Lactobacillus

Int J Syst Evol Microbiol, 2005 Jan, 55(Pt 1), 401 - 4
Lactobacillus delbrueckii subsp . indicus subsp . nov., isolated from Indian dairy products; Dellaglio F et al.; Four strains isolated from Indian dairy products and initially identified as Lactobacillus delbrueckii could not be assigned to a definite subspecies because molecular identification and phenotypic traits did not agree with those of recognized subspecies of L . delbrueckii . Hybridization of total DNA (78-86 % against type strains of the other three subspecies), AFLP and RAPD-PCR fingerprints, phylogenetic analysis based on 16S rRNA gene sequences and sequence analysis of two coding genes (recA and hsp60), together with phenotypic profiles, indicated that the four strains form a coherent cluster and represent a novel subspecies, for which the name Lactobacillus delbrueckii subsp . indicus subsp . nov . is proposed . The type strain is NCC725(T) (=LMG 22083(T)=DSM 15996(T)).

Int J Syst Evol Microbiol, 2005 Jan, 55(Pt 1), 197 - 207
Polyphasic study of wine Lactobacillus strains: taxonomic implications; Rodas AM et al.; One hundred and seventy-eight lactobacilli isolated from wine were characterized by a polyphasic approach . Strains were phenotypically identified at genus and species level by classical tests including the analysis of cell morphology, homo/heterofermentative character, sugar fermentation patterns, growth at different temperatures and the optical nature of the isomer of lactic acid produced from glucose . Molecular techniques such as random amplification of polymorphic DNA (RAPD), amplified 16S rDNA restriction analysis (16S-ARDRA), PFGE-RFLP and ribotyping were used to characterize strains, and their potential for identification and/or typing was evaluated . The information obtained with these techniques was processed with the BioNumerics software in order to analyse relationships existing between isolated strains and various reference species of the genus . Then, taxonomic dendrograms were obtained, and this information allowed the proposal of molecular procedures suitable for the identification and typing of these wine micro-organisms . The techniques useful for both identification and typing were RAPD and ribotyping, while 16S-ARDRA was only useful for identification and PFGE-RFLP only for typing purposes . The wine strains were identified as Lactobacillus brevis (19 strains), Lactobacillus collinoides (2 strains), Lactobacillus hilgardii (71 strains), Lactobacillus paracasei (13 strains), Lactobacillus pentosus (2 strains), Lactobacillus plantarum (34 strains) and Lactobacillus mali (10 strains).

Int J Syst Evol Microbiol, 2005 Jan, 55(Pt 1), 83 - 5
Lactobacillus satsumensis sp . nov., isolated from mashes of shochu, a traditional Japanese distilled spirit made from fermented rice and other starchy materials; Endo A et al.; Gram-positive, rod-shaped, motile lactic acid bacteria (strains NRIC 0603, NRIC 0604(T), NRIC 0605 and NRIC 0606) were isolated from shochu mashes using an enrichment culture approach . These strains clustered in the Lactobacillus casei-Pediococcus group and were closely related to Lactobacillus nagelii and Lactobacillus mali on the basis of 16S rRNA gene sequence similarity . Levels of DNA-DNA relatedness revealed genotypic separation of the four isolates from the above two species . The isolates are therefore considered to represent a novel species, for which the name Lactobacillus satsumensis is proposed . The type strain is NRIC 0604(T) (=JCM 12392(T)=DSM 16230(T)).

Int J Syst Evol Microbiol, 2005 Jan, 55(Pt 1), 77 - 82
Lactobacillus gastricus sp . nov., Lactobacillus antri sp . nov., Lactobacillus kalixensis sp . nov . and Lactobacillus ultunensis sp . nov., isolated from human stomach mucosa; Roos S et al.; In an attempt to study the composition of the Lactobacillus flora from mucosa of human stomach, 16S rRNA gene sequences of 129 isolates were analysed . For 15 of these, the results differed significantly from known sequences, and additional tests were performed to determine whether these isolates represented as yet unrecognized species . Phenotypic and genetic characteristics revealed that these isolates represented four novel Lactobacillus species . Two belong to the Lactobacillus reuteri and the other two to the Lactobacillus delbrueckii subgroup of Lactobacillus . The names Lactobacillus gastricus sp . nov., Lactobacillus antri sp . nov., Lactobacillus kalixensis sp . nov . and Lactobacillus ultunensis sp . nov . are proposed, with the respective type strains Kx156A7(T) (=LMG 22113(T)=DSM 16045(T)=CCUG 48454(T)), Kx146A4(T) (=LMG 22111(T)=DSM 16041(T)=CCUG 48456(T)), Kx127A2(T) (=LMG 22115(T)=DSM 16043(T)=CCUG 48459(T)) and Kx146C1(T) (=LMG 22117(T)=DSM 16047(T)=CCUG 48460(T)).

Int J Syst Evol Microbiol, 2005 Jan, 55(Pt 1), 35 - 40
Lactobacillus rossii sp . nov., isolated from wheat sourdough; Corsetti A et al.; Screening of sourdough lactic acid bacteria for bacteriocin production resulted in the isolation of a Gram-positive, catalase-negative, non-spore-forming, non-motile rod bacterium (strain CS1(T)) that could not be associated with any previously described species . Comparative 16S rRNA gene sequence analysis recognized strain CS1(T) as a distinct member of the genus Lactobacillus . By a species-specific PCR strategy, five additional strains previously isolated from sourdoughs were found to belong to the same species as strain CS1(T), as confirmed by 16S rRNA gene sequence analysis . The closest related species were Lactobacillus durianis, Lactobacillus malefermentans and Lactobacillus suebicus, with which strain CS1(T) shared 93 % sequence similarity . For a further characterization of strain CS1(T), physiological (growth temperature, CO(2) production, hydrolysis of arginine, isomeric type of lactate, sugar fermentation) and chemotaxonomic (G+C content and peptidoglycan structure) properties were determined . Phenotypic characterization showed that strain CS1(T) was a member of the obligately heterofermentative group of the genus Lactobacillus . The DNA G+C content was 44.6 mol% . The peptidoglycan was of the A3alpha (l-lys-l-ser-l-Ala(2)) type . Physiological, biochemical and genotypic data, as well as results of DNA-DNA hybridization of genomic DNA with one of the closest phylogenetic relatives, L . durianis (34.3 %), indicated that strain CS1(T) represents a novel species of the genus Lactobacillus for which the name Lactobacillus rossii sp . nov . is proposed . The type strain of this species is CS1(T) (=ATCC BAA-822(T)=DSM 15814(T)).

Int J Syst Evol Microbiol, 2005 Jan, 55(Pt 1), 31 - 4
Lactobacillus suntoryeus sp . nov., isolated from malt whisky distilleries; Cachat E et al.; Eight strains of Lactobacillus with identical partial 16S rRNA gene sequences and similar randomly amplified polymorphic DNA patterns were isolated from fermentation samples from Japanese and Scottish malt whisky distilleries . Phylogenetic analysis of almost complete 16S rRNA gene sequences from three representative strains (two from Japan, one from Scotland) placed them in the genus Lactobacillus as members of the Lactobacillus acidophilus group . Lactobacillus helveticus and Lactobacillus gallinarum were the most closely related species, with 16S rRNA gene similarities of 99.3 and 98.1 %, respectively . A similar phylogeny was derived from partial sequences of elongation factor Tu (tuf) genes in which the alleles from the three distillery isolates were identical and shared 99.0 % similarity with L . helveticus and L . gallinarum tuf genes . S-layer (slp) gene sequences suggested different relationships among the strains and the distillery isolates no longer formed a monophyletic group . The alleles from the Japanese and Scottish strains shared only 54 % similarity . Chromosomal DNA from the distillery strains gave DNA-DNA hybridization values between 79 and 100 % but showed less than 43 and 22 % reassociation with L . helveticus and L . gallinarum DNA, respectively . The name Lactobacillus suntoryeus sp . nov . is proposed for this novel taxon; the type strain is strain SA(T) (=LMG 22464(T)=NCIMB 14005(T)).

J Dairy Sci, 2005 Feb, 88(2), 527 - 33
Clinical Effects of Lactobacillus acidophilus Strain L-92 on Perennial Allergic Rhinitis: A Double-Blind, Placebo-Controlled Study; Ishida Y et al.; Studies in animals have suggested that lactic acid bacteria alleviate allergic diseases, however, little information is available on their clinical effect on allergy in humans . Thus, we examined the efficacy of orally administered Lactobacillus acidophilus strain L-92 (L-92) on perennial allergic rhinitis . In a randomized, double-blind, placebo-controlled clinical trial, 49 patients with perennial allergic rhinitis were randomized to receive either 100 mL of heat-treated fermented milk containing L-92 (n = 25) or acidified milk without lactic acid bacteria (placebo; n = 24) for 8 wk . The severity of symptoms was evaluated based on the changes in the scores of clinical symptoms . Oral administration of milk fermented with L-92 resulted in a statistically significant improvement of nasal symptom-medication scores . Ocular symptom-medication scores of patients in the L-92 intervention group tended to improve compared with those in the placebo group . In addition, clear decreases of the scores of swelling and color of the nasal mucosa were observed in the L-92 intervention group at 6 and 8 wk after the start of ingestion of fermented milk . There were no significant differences in serum antihouse dust mite immunoglobulin E levels nor in T helper type 1/T helper type 2 ratio between the 2 groups . These results suggest that oral administration of L-92 can alleviate the symptoms of perennial allergic rhinitis, however, statistically significant changes were not shown in blood parameters.

J Microbiol, 2004 Dec, 42(4), 315 - 8
Probiotication of tomato juice by lactic Acid bacteria; Yoon KY et al.; This study was undertaken to determine the suitability of tomato juice as a raw material for production of probiotic juice by four lactic acid bacteria (Latobacillus acidophilus LA39, Lactobacillus plantarum C3, Lactobacillus casei A4, and Lactobacillus delbrueckii D7) . Tomato juice was inoculated with a 24-h-old culture and incubated at 30 degrees C . Changes in pH, acidity, sugar content, and viable cell counts during fermentation under controlled conditions were measured . The lactic acid cultures reduced the pH to 4.1 or below and increased the acidity to 0.65% or higher, and the viable cell counts (CFU) reached nearly 1.0 to 9.0x10(9)/ml after 72 h fermentation . The viable cell counts of the four lactic acid bacteria in the fermented tomato juice ranged from 10(6) to 10(8) CFU/ml after 4 weeks of cold storage at 4 degrees C . Probiotic tomato juice could serve as a health beverage for vegetarians or consumers who are allergic to dairy products.

J Microbiol Methods, 2005 Mar, 60(3), 353 - 63
Modified electroporation protocol for Lactobacilli isolated from the chicken crop facilitates transformation and the use of a genetic tool; Mason CK et al.; Isolates of Lactobacillus spp . from a collection of potentially probiotic strains isolated from the crops of broiler chickens were found to be non-electrotransformable using published techniques . One strain of Lactobacillus salivarius was shown to develop electrocompetence when an overnight culture was incubated in fresh medium . The effect was enhanced if glycine was incorporated into the fresh growth medium . When these modifications were applied to a number of other crop isolates of Lactobacillus spp., electrocompetence could be detected in approximately half the strains tested . Two temperature sensitive plasmid vectors that had been used for the genetic modification of other lactic acid bacteria were introduced into a crop strain of Lb . salivarius . Both showed temperature sensitivity at 42 degrees C and above but were relatively stable at 37 degrees C . The genetic tool harbouring an IS element allowed the delivery of the plasmid to multiple independent sites in the host chromosome . Harnessing such genetic tools will facilitate the future genetic analysis of the host bacterium.

Microbiol Res, 2004, 159(4), 339 - 46
Potential of selected lactic acid bacteria to produce food compatible antifungal metabolites; De Muynck C et al.; The aim of this study was to assess the potential of lactic acid bacteria to inhibit the outgrowth of some common food-spoiling fungi . Culture supernatants of 17 Lactic acid bacterial strains as well as of three commercial probiotic cultures were evaluated for antifungal activity using an agar-diffusion method . The method parameters were chosen in order to reveal compounds for potential use in food (bio)preservation . Thirteen strains showed antifungal activity of which five strains were very promising: Lactobacillus acidophilus LMG 9433, L . amylovorus DSM 20532, L . brevis LMG 6906, L . coryniformis subsp . coryniformis LMG 9196 and L . plantarum LMG 6907 . Four of these five strains were further examined; it was found that the produced antifungal metabolites were pH-dependent . The exact chemical nature of these substances has not been revealed yet.

Oral Health Prev Dent, 2003, 1(2), 157 - 62
Influence of dental restorative materials on salivary Streptococcus mutans and lactobacilli in the primary dentition; Willershausen B et al.; PURPOSE: When restoring deciduous teeth with extensive lesions, preformed stainless steel crowns are increasingly used in addition to resin-based materials . The aim of the present clinical study was to examine the influence of composites and stainless steel crowns on the cariogenic bacteria S . mutans and Lactobacilli by means of a commercially available salivary test (CRT bacteria) in 100 children . MATERIALS AND METHODS: For 50 children (30 boys, 20 girls, mean age 5.7 +/- 2.4 years) only composites have been used as a restorative material (an average of 9 fillings per child, total number: n=450), which had been in situ for an average of 15.9 months at the time of the examination . In another group of 50 children (33 boys, 17 girls, mean age 5.9 +/- 2.5 years), stainless steel crowns and composites have been used for restorations (an average of 3.5 stainless steel crowns per child, total number: n=174, an average of 7 composite fillings per child, total number n=348), which had been in place for a mean period of 17.2 months . RESULTS: The salivary examination of the children with composite restorations showed a high proportion of patients (64%, n=32) with high numbers (> or = 10(5)) of S . mutans and Lactobacilli (54%, n=27) . Only 22% (n=11) of the children with additional stainless steel crown restorations were found to have high numbers (> or = 10(5)) of S . mutans and Lactobacilli (34%, n=17) . CONCLUSION: This study shows a potential positive inhibitory effect of stainless steel crown restorations as compared to composite fillings with respect to the oral bacterial colonization.

Oral Health Prev Dent, 2003, 1(3), 185 - 94
A ten-year cross-sectional and follow-up study of salivary flow rates and mutans streptococci and lactobacillus counts in elderly Swedish individuals; Fure S; PURPOSE: The whole salivary flow rates and microbial conditions were examined in a 10-year follow-up and cross-sectional study in a random sample of elderly inhabitants of Gothenburg . MATERIAL AND METHODS: Of the 208 individuals examined at baseline, forty-nine per cent participated in the follow-up, 56, 37 and nine persons, respectively, in the age groups of 65, 75 and 85 years . In addition, a new random sample of 98 individuals aged 55 year was examined . RESULTS: The mean secretion rate decreased significantly with increasing age in terms of unstimulated and stimulated whole saliva . Of the 200 participants, 50% were taking medication, which could have hyposalivatory side-effects . Persons with a daily intake of > or =4 drugs had significantly lower unstimulated and stimulated secretion rates . Forty-five persons reported subjective dryness in the mouth . The mean saliva secretion rates among these persons were significantly lower and the number of drugs consumed significantly higher than in people with no such complaints . The overall salivary counts of lactobacilli and mutants streptococci increased with age . Higher counts of these bacteria were found in persons wearing removable dentures than in persons without dentures . CONCLUSION: The salivary and microbial conditions ought to be continuously monitored in old people, in order to identify those who need oral health promotive measures.

Appl Environ Microbiol, 2005 Jan, 71(1), 297 - 302
Starch-Binding Domain Affects Catalysis in Two Lactobacillus {alpha}-Amylases; Rodriguez-Sanoja R et al.; A new starch-binding domain (SBD) was recently described in alpha-amylases from three lactobacilli (Lactobacillus amylovorus, Lactobacillus plantarum, and Lactobacillus manihotivorans) . Usually, the SBD is formed by 100 amino acids, but the SBD sequences of the mentioned lactobacillus alpha-amylases consist of almost 500 amino acids that are organized in tandem repeats . The three lactobacillus amylase genes share more than 98% sequence identity . In spite of this identity, the SBD structures seem to be quite different . To investigate whether the observed differences in the SBDs have an effect on the hydrolytic capability of the enzymes, a kinetic study of L . amylovorus and L . plantarum amylases was developed, with both enzymes acting on several starch sources in granular and gelatinized forms . Results showed that the amylolytic capacities of these enzymes are quite different; the L . amylovorus alpha-amylase is, on average, 10 times more efficient than the L . plantarum enzyme in hydrolyzing all the tested polymeric starches, with only a minor difference in the adsorption capacities.

J Biotechnol, 2005 Feb 23, 115(4), 405 - 12 Epub 2004 Nov 11.
High-pressure inactivation of Saccharomyces cerevisiae and Lactobacillus plantarum at subzero temperatures; Perrier-Cornet JM et al.; High hydrostatic pressure is a new technology in the food processing industry, and is used for cold pasteurization of food products . However, the pressure inactivation of food-borne microorganisms requires very high pressures (generally more than 400MPa) and long pressure holding times (5min or more) . Carrying out pressure processing at low temperatures without freezing can reduce these parameters, which presently limit the application of this technology, in keeping the quality of fresh raw product . The yeast, Saccharomyces cerevisiae and the bacterium, Lactobacillus plantarum were pressurized for 10min at temperatures between -20 and 25 degrees C and pressure between 100 and 350MPa . Pressurization at subzero temperatures without freezing significantly enhanced the effect of pressure . For example, at a pressure of 150MPa, the decrease in temperature from ambient to -20 degrees C allowed an increase in the pressure-induced inactivation from less than 1log up to 7-8log for each microorganism studied . However, for comparable inactivation levels, the kinetics of microorganism inactivation did not differ, which suggests identical inactivation mechanisms . Implications of water thermodynamical properties like compression, protein denaturation, as well as membrane phase transitions, are discussed.

Reprod Nutr Dev, 2004 Sep-Oct, 44(5), 449 - 57
Effects of the administration of Lactobacilli on body growth and on the metabolic profile in growing Maltese goat kids; Chiofalo V et al.; The aim of this study was to evaluate the effect of some lactobacilli on body growth and on the metabolic-nutritional status in growing goat kids . Twenty growing Maltese goat kids (10 Control and 10 Bios) were studied . The animals of the Bios group received a concentrate including 1 g x kg(-1) of SEB Bovino (spray-dried), Akron S.r.l., Italy, with non bacterial components: gum arabic, soybean meal, silicate alum of magnesium, and with bacterial components: 10(11) cfu kg(-1) each of Lactobacillus acidophilus, Lactobacillus salivarius, Lactobacillus reuteri . Monthly, bio-metric and weight evaluations were carried out on each animal and individual blood samples were taken . The Bios group showed the highest body weight (Control 19 vs . Bios 23 kg P < 0.001), anamorphosis (Control 71 vs . Bios 78 P < 0.05) and body proportion (Control 35 vs . Bios 41 P < 0.001) indices; the lowest levels of Non Esterified Fatty Acids (Control 0.778 vs . Bios 0.403 mmol L(-1) P < 0.001), triglycerides (Control 0.21 vs . Bios 0.18 mmol L(-1) P < 0.05), urea (Control 8.83 vs . Bios 7.65 mmol L(-1) P < 0.05) and the highest levels of Alkaline Phosphatase (Control 270 vs . Bios 851 U L(-1) P < 0.01) and Creatine Kinase (Control 173 vs . Bios 285 U L(-1) P < 0.01) . The results testify to the better metabolic activity of the Bios group which achieved, at the end of the trial (7 months old), about 99% of the morphological development of the adult, therefore an adequate structure for mating and going into production within the first year of life.

Reprod Nutr Dev, 2004 Sep-Oct, 44(5), 397 - 405
Preliminary assays for the development of a probiotic for goats; Draksler D et al.; In order to determine probiotic properties, 137 strains of lactic acid bacteria from the feces of Creole goats were screened, only six were resistant to pH 2.0 and bile salts (0.3%) . Three strains identified as Lactobacillus and two as Enterococcus showed agglutination with the treated yeast . Between them, Lactobacillus DDL17, DDL19, DDL48 and Enterococcus DDE39 demonstrated high specificity in this test because the correspondent agglutination was inhibited by one sugar, suggesting the presence of a lectin-like structure in their cell walls, which could be due to adhesion ability . Another Enterococcus strain (DDE55) showed low affinity because five sugars inhibited the agglutination of the treated yeasts . The results of hydrophobic properties showed that the strains who were able to agglutinate yeasts presented similar hydrophobic characteristics as hexadecane, xylene and toluene, but high specificity was not related to a high hydrophobicity . Only two strains (Lactobacillus DDL19 and DDL48) showed aggregation with the lowest concentration of ammonium sulfate, complementing the hydrophobicity assay . Only one strain, Lactobacillus DDL48, showed an inhibition against an enteric indicator strain (Salmonella Typhimurium and Escherichia coli O111) . This inhibitory action was not affected by the addition of catalase and no inhibition was detected after neutralizing the supernatant culture fluid . These strains could be pre-selected in order to complete studies focused on designing a probiotic for use in goat feed.

Zh Mikrobiol Epidemiol Immunobiol, 2004 Nov-Dec, (6), 101 - 6
{The role of cytokines in the immunoreactivity modulation with bacteria of the Lactobacillus genus}
{Modulation of immune system cells by lactobacilli}
{No authors listed}

The influence of lactobacilli on the proliferative potential of immune system cells after the intragastral administration of viable microbial cells and their native filtrates to mice CBA is evaluated . The data have been obtained on the modulating influence of lactobacilli on the formation of T- and B-cell immune response--their role in maintaining homeostatsis and specific features of cell mediated immune reactions after the intragastral administration of virulent Shigella dysenteriae for modeling experimental infection in CBA mice . The mechanisms of the immunomodulating action of lactobacilli on local and systemic reactions of the host as well as realization of the protective properties of lactobacilli against the causative agents of acute enteric infections are discussed.

Appl Microbiol Biotechnol . 2005 Jan 6; {Epub ahead of print}
Cell surface display system for Lactococcus lactis: a novel development for oral vaccine; Raha AR et al.; The food-grade Lactococcus lactis is a potential vector to be used as a live vehicle for the delivery of heterologous proteins for vaccine and pharmaceutical purposes . We constructed a plasmid vector pSVac that harbors a 255-bp single-repeat sequence of the cell wall-binding protein region of the AcmA protein . The recombinant plasmid was transformed into Escherichia coli and expression of the gene fragment was driven by the T7 promoter of the plasmid . SDS-PAGE showed the presence of the putative AcmA' fragment and this was confirmed by Western blot analysis . The protein was isolated and purified using a His-tag affinity column . When mixed with a culture of L . lactis MG1363, ELISA and immunofluorescence assays showed that the cell wall-binding fragment was anchored onto the outer surface of the bacteria . This indicated that the AcmA' repeat unit retained the active site for binding onto the cell wall surface of the L . lactis cells . Stability assays showed that the fusion proteins (AcmA/A1, AcmA/A3) were stably docked onto the surface for at least 5 days . The AcmA' fragment was also shown to be able to strongly bind onto the cell surface of naturally occurring lactococcal strains and Lactobacillus and, with less strength, the cell surface of Bacillus sphericus . The new system designed for cell surface display of recombinant proteins on L . lactis was evaluated for the expression and display of A1 and A3 regions of the VP1 protein of enterovirus 71 (EV71) . The A1 and A3 regions of the VP1 protein of EV71 were cloned upstream to the cell wall-binding domains of AcmA protein and successfully expressed as AcmA/A1 and AcmA/A3 . Whole-cell ELISA showed the successful display of VP1 protein epitopes of EV71 on the surface of L . lactis . The success of the anchoring system developed in this study for docking the A1 and A3 epitopes of VP1 onto the surface of L . lactis cells opens up the possibilities of peptide and protein display for not only Lactococcus but also for other gram-positive bacteria . This novel way of displaying epitopes on the cell surface of L . lactis and other related organisms should be very useful in the delivery of vaccines and other useful proteins.

Pol J Vet Sci, 2004, 7(4), 251 - 9
Effect of chosen lactic acid bacteria strains on Staphylococcus aureus in vitro as well as in meat and raw sausages; Gomolka-Pawlicka M et al.; The present study was aimed at determining the influence of 15 strains of lactic acid bacteria on the growth of 2 Staphylococcus aureus strains in vitro as well as in meat and raw sausages . The investigations were performed within the framework of three alternate stages which differed in respect to the products studied, the number of Lactobacillus sp . strains and, partly, methodological approach . The study also considered water activity (a(w)) and pH of the products investigated . The results obtained are demonstrated in 5 diagrams . It was found that among 15 strains of Lactobacillus aureus investigated only one strain, Lactobacillus helveticus T 78, showed antagonistic effect on studied strains of Staphylococcus aureus both in vitro as well as in meat and raw sausages . Five other strains of Lactobacillus spp . displayed the antagonistic effect in vitro only . The temperature and incubation time of sausages, but also the type of sausage stuffing were found to have a distinct or slight influence, respectively, on the antagonistic interaction between the bacteria . However, this phenomenon was affected by neither a(w) nor pH.

J Food Prot, 2004 Dec, 67(12), 2779 - 85
Interrelationships among microbiological, physicochemical, and biochemical properties of Terrincho cheese, with emphasis on biogenic amines; Pinho O et al.; Changes in the microbiological, physicochemical, and biochemical characteristics of Terrincho cheese as represented by native microflora, pH, water activity, soluble nitrogen fractions, free amino acids, and biogenic amines (e.g., ethylamine, dimethylamine, tryptamine, phenylethylamine, putrescine, cadaverine, histamine, tyramine, cystamine, and spermine) during ripening were monitored . Terrincho is a traditional Portuguese cheese manufactured from raw ewe's milk . The main groups of microorganisms (lactococci, lactobacilli, enterococci, pseudomonads, staphylococci, coliforms, yeasts, and molds) were determined following conventional microbiological procedures . Free amino acids and biogenic amines were determined by reverse-phase high-performance liquid chromatography, following extraction from the cheese matrix and derivatization with dabsyl chloride . The total content of free amino acids ranged from 1,730 mg/kg of dry matter at the beginning of the ripening stage to 5,180 mg/kg of dry matter by day 60 of ripening; such an increase was highly correlated with the increase of water-soluble nitrogen in total nitrogen, 12% trichloroacetic acid-soluble nitrogen in total nitrogen, and 5% phosphotungstic acid-soluble nitrogen in total nitrogen throughout ripening . Histamine was consistently present at very low levels, whereas putrescine, cadaverine, and tryptamine were the dominant biogenic amines and increased in concentration during ripening . Ethylamine, tryptamine, phenylethylamine, and cystamine reached maxima by 30 days of ripening and decreased thereafter . Significant correlations between amino acid precursors and corresponding biogenic amines, as well as between biogenic amines and microbial viable numbers, were observed.

J Food Prot, 2004 Dec, 67(12), 2727 - 34
Characterization and antimicrobial activity of bacteriocin 217 produced by natural isolate Lactobacillus paracasei subsp . paracasei BGBUK2-16; Lozo J et al.; The strain Lactobacillus paracasei subsp . paracasei BGBUK2-16 . which was isolated from traditionally homemade white-pickled cheese, produces bacteriocin 217 (Bac217; approximately 7 kDa) . The onset of Bac217 biosynthesis was observed in the logarithmic phase of growth, and the production plateau was reached after 9 or 12 h of incubation at 37 and 30 degrees C, respectively, when culture entered the early stationary phase . Biochemical characterization showed that Bac217 retained antimicrobial activity within the range of pH 3 to 12 or after treatment at 100 degrees C for 15 min . Bac217 antimicrobial activity also remained unchanged after storage at 4 degrees C for 6 months or -20 degrees C for up to 12 months . However, Bac217 activity was completely lost after treatment with different proteolytic enzymes . BGBUK2-16 contains only one plasmid about 80 kb in size . Plasmid curing indicated that genes coding for Bac217 synthesis and immunity seem to be located on this plasmid . Bac217 exhibited antimicrobial activity against some pathogenic bacteria, such as Staphylococcus aureus and Bacillus cereus . Interestingly, Bac217 showed activity against Salmonella sp . and Pseudomonas aeruginosa ATCC27853 . The inhibitory effect of BGBUK2-16 on the growth of S . aureus in mixed culture was observed . S . aureus treatment with Bac217 led to a considerable decrease (CFU/ml) within a short period of time . The mode of Bac217 action on S . aureus was identified as bactericidal . It should be noted that the strain BGBUK2-16 was shown to be resistant to bacteriocin nisin, which is otherwise widely used as a food additive for fermented dairy products.

Home Healthc Nurse, 2005 Jan, 23(1), 13 - 15
Bugs, Drugs, and Yogurt; Neafsey PJ et al.; Mrs . S . is suffering from a yeast infection after a course of antibiotics . Despite several unsuccessful self-treatments with an over-the-counter intravaginal clotrimazole treatment, she remained symptomatic and she was prescribed oral Diflucan (fluconazole) . At first it seems that Mrs . S . has been cured, but a few days later, her symptoms return . You remember reading an article on Lactobacillus acidophilus as a remedy for yeast infections . Is there any validity to this treatment or is it just the latest craze of naturopathic medicine?

Appl Microbiol Biotechnol . 2005 Jan 4; {Epub ahead of print}
Lactobacillus reuteri ATCC 53608 mdh gene cloning and recombinant mannitol dehydrogenase characterization; Sasaki Y et al.; A gene encoding mannitol-2-dehydrogenase (E.C . 1.1.1.138) (MDH) was cloned from Lactobacillus reuteri and expressed in Escherichia coli . The 1,008-bp gene encodes a protein consisting of 336 amino acids, with a predicted molecular mass of 35,920 Da . The deduced amino acid sequence of L . reuteri MDH (LRMDH) is 77% and 76% similar to the MDHs from Leuconostoc mesenteroides and Leuconostoc pseudomesenteroides, respectively . The purified recombinant enzyme appears as a single band of 40 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but gel filtration indicates that the native enzyme is a dimer . The optimum temperature for the recombinant enzyme is 37 degrees C, the pH optima for D: -fructose reduction and D: -mannitol oxidation are 5.4 and 6.2, respectively . The K(m) values for NAD (9 mM) and NADH (0.24 mM) are significantly higher than those for NADP (0.35 mM) and NADPH (0.04 mM) . The K(m) values of LRMDH for D: -fructose and D: -mannitol are 34 mM and 54 mM, respectively . Contrary to what the enzyme sequence suggests, recombinant LRMDH contains a single catalytic zinc per subunit.

Biofactors, 2004, 22(1-4), 63 - 6
Antitumor activity and action mechanisms of Lactobacillus casei through the regulation of immune responses; Matsuzaki T et al.; Lactobacillus casei strain Shirota (LcS) has been demonstrated to have beneficial effects in numerous disease models, especially murine autoimmune disease and carcinogenesis models, via host immune modulation . It has also been reported that LcS induced recovery of the host immune responses that were decreased by treatment with carcinogens, and that augmented the natural killer activity and T cell functions of host immune cells . After LcS is ingested by the host, it is incorporated into M cells in Peyer's patches (PP) and digested to form active components . In PP, macrophages or dendritic cells that phagocytosed LcS gained ability to produce tumor necrosis factor-alpha . The components of LcS digested in PP were then recognized through toll-like receptor 2 in macrophages, resulting in the production of several cytokines which elicit varied responses in host immune cells . Also, it was observed by 2D-PAGE analyses that the expression level and/or the phosphorylation of some proteins in PP and mesenteric lymph nodes were definitely altered after the ingestion of LcS, providing more evidence of cellular responses.

Biofactors, 2004, 21(1-4), 155 - 8
Isolation and identification of lactic acid bacteria with effect of immune protection to Eschericia coli in mice; Ishida-Fujii K et al.; Lactic acid bacteria were isolated from an alcohol fermentation broth, and the activity as a probiotic was examined using pathogenic E . coli . Thirty-six strains exhibiting good growth were isolated in the medium of concentrated mush which was a residue resulted in the alcohol distillation process . One of these strains, Lactobacillus paracasei subsp . paracasei I-5, could be grown in the medium containing 8 vol% ethanol and at 45 degrees C . The characteristics were different from the type strain, L . paracasei subsp . paracasei NBRC 15889trade mark . L . paracasei I-5 showed an excellent growth in the concentrated mush, which just diluted two-fold and adjusted the pH . ICR mice were fed with a standard germ-free feed (CMF) and the strain I-5 (7x10(9) cells/day) was orally administrated for 11 days prior to the intraperitoneal challenge with pathogenic E . coli Juhl . After the challenge, mice administrated the strain I-5 exhibited a high survival rate and survival extension days (p < 0.01) compared with the control . The results suggested that the strain might enhance the animal resistance against microbial pathogens . Neonatal diarrhea caused by E . coli is a serious disease in calf breeding . The strain might be practically valuable to prevent diarrhea in calves.

Pediatrics, 2005 Jan, 115(1), 178 - 81
Lactobacillus sepsis associated with probiotic therapy; Land MH et al.; Probiotic strains of lactobacilli are increasingly being used in clinical practice because of their many health benefits . Infections associated with probiotic strains of lactobacilli are extremely rare . We describe 2 patients who received probiotic lactobacilli and subsequently developed bacteremia and sepsis attributable to Lactobacillus species . Molecular DNA fingerprinting analysis showed that the Lactobacillus strain isolated from blood samples was indistinguishable from the probiotic strain ingested by the patients . This report indicates, for the first time, that invasive disease can be associated with probiotic lactobacilli . This report should not discourage the appropriate use of Lactobacillus or other probiotic agents but should serve as a reminder that these agents can cause invasive disease in certain populations.

Vaccine, 2005 Jan 19, 23(9), 1126 - 34
Intranasal immunisation using recombinant Lactobacillus johnsonii as a new strategy to prevent allergic disease; Scheppler L et al.; We have previously demonstrated the induction of a specific anti-IgE response in vivo by parenteral immunisation of rhesus monkeys using short IgE mimotopes or an anti-idiotypic antibody mimicking an IgE epitope . Such specific anti-IgE responses may be of clinical benefit for atopic patients . In this study, we examined the potential for a more convenient therapy via mucosal immunisation using live recombinant Lactobacillus johnsonii (Lj) as a vaccine delivery vehicle . Either an anti-idiotypic scFv or an IgE mimotope were expressed on the surface of Lj as fusion proteins using the cell wall anchored proteinase PrtB from Lactobacillus delbrueckii subsp . bulgaricus . The recombinant Lj were shown to express the heterologous fusion proteins and were specifically recognised by the corresponding anti-human IgE monoclonal antibody . Subcutaneous and intranasal immunisation of mice with recombinant Lj, expressing these fusion proteins induced a systemic IgG response against human IgE . Our data suggest that recombinant Lactobacilli expressing IgE epitopes may represent a novel means of vaccination to induce a beneficial anti-IgE response.

Biopolymers . 2004 Dec 29;77(2):98-106 {Epub ahead of print}
Viscometric and static light scattering studies on an exopolysaccharide produced by Lactobacillus delbrueckii subspecies bulgaricus NCFB 2483; Goh KK et al.; The rheological properties and molecular parameters of a purified exopolysaccharide (EPS) produced by a ropy strain of Lactobacillus delbrueckii subsp . bulgaricus NCFB 2483 were investigated . Using capillary viscometry, an intrinsic viscosity of 2,013 mL/g was obtained . The flow curves were fitted by both the Carreau and the Cross equations for shear-thinning fluids, with the Carreau equation giving a better fit . The Cross equation fitted fairly well the plot of reduced viscosity as a function of reduced shear rate with an exponent value (1 - n) of approximately 0.76, typical of random coil polymers . Furthermore, the concentration dependence of the viscosity plot showed a gradient of approximately 1.1 in the dilute regime and 3.3 in the semidilute regime . Molecular parameters were obtained using a multiangle laser light scattering technique . The 2483 EPS molecules had a weight-average molar mass of approximately 2 x 10(6) Da and a z-average root mean square radius (RMS) of approximately 151 nm . From the light scattering data, the bacterial EPS was also found to have a low polydispersity index ( approximately 1.15) and adopt a random coil conformation . (c) 2004 Wiley Periodicals, Inc . Biopolymers, 2005.

Nutr Clin Care, 2004 Jul-Sep, 7(3), 104 - 13
Probiotics and prebiotics in clinical practice; Broussard EK et al.; Probiotics are living organisms that have a beneficial therapeutic effect on the host when ingested . The bacteria Lactobacillus GG and the yeast Saccharomyces boulardii have been shown in controlled trials to prevent antibiotic-associated diarrhea . Prebiotics are non-living, indigestible polysaccharides metabolized in the intestine that induce physical and bacteriologic musocal modifications . Rather than introducing exogenous bacteria into the intestinal millieu, prebiotics stimulate certain indigenous bacteria already resident in the gut.

Biosci Biotechnol Biochem, 2004 Dec, 68(12), 2427 - 35
Isolation of Lactobacillus sakei Strain KJ-2008 and Its Removal of Characteristic Malodorous Gases under Anaerobic Culture Conditions; Kim JD et al.; A number of different sources, such as composts, leachates, and pig feces samples were collected from different pig farms in Korea . Several microorganisms were screened for their ability to deodorize the malodorous gases . As a result, a novel malodorous gas-deodorizing bacterial strain KJ-2008 was isolated due to the most abundant of nitrate-supplemented minimal media under anaerobic conditions . Crimp-sealed serum bottles containing nitrate-supplemented minimal medium (MM-NO(3)(-)) in airtight conditions were inoculated with KJ-2008 . Nitrate concentration decreased rapidly after 20 h incubation and nitrite production reached almost zero during the time the experimental was carried out . Taxonomic identification including 16S rDNA base sequencing and phylogenetic analysis indicated that the isolate KJ-2008 had a 99.8% homology in its 16S rDNA base sequence with Lactobacillus sakei . Among the volatile fatty acids, acetic acid contained in large amounts in fresh piggery slurry decreased about 40% after 50 h incubation of the strain KJ-2008 . n-Butyric acid, n-valeric acid, and iso-valeric acid gradually decreased, and iso-butyric acid and capronic acid dramatically eliminated at initial time with the treatment . Moreover, NH(3) removal efficiency reached a maximum of 98.5% after 50 h of incubation . The concentration of H(2)S did not change.

Infect Immun, 2005 Jan, 73(1), 476 - 84
CCL20/macrophage inflammatory protein 3alpha and tumor necrosis factor alpha production by primary uterine epithelial cells in response to treatment with lipopolysaccharide or Pam3Cys; Crane-Godreau MA et al.; Having previously shown that CCL20/macrophage inflammatory protein 3alpha and tumor necrosis factor alpha (TNF-alpha) are released by polarized primary rat uterine epithelial cells (UEC) in response to Escherichia coli but not to Lactobacillus rhamnosus, we sought to determine if epithelial cells are responsive to pathogen-associated molecular patterns (PAMP), including lipopolysaccharide (LPS), lipoteichoic acid (LTA), and Pam(3)Cys, a bacterial lipoprotein analog . Epithelial cells were grown to confluence on Nunc cell culture inserts prior to apical treatment with PAMPs . In response to LPS, LTA, and Pam(3)Cys (EMC Microcollection GmbH, Tubingen, Germany), CCL20 levels increased (4- to 10-fold) while PAMPs caused increased TNF-alpha (1- to 4-fold) in the medium collected after 24 h of incubation . Both apical and basolateral secretion of CCL20 and TNF-alpha increased in response to PAMPs, but treatments had no effect on cell viability and integrity, as measured by transepithelial resistance . Time course studies of CCL20 and TNF-alpha release in response to Pam(3)Cys and LPS indicated that CCL20 release peaked between 2 and 4 h after treatment, whereas TNF-alpha release was gradual over the length of the incubation . Freeze-thaw and cell lysis experiments, along with actinomycin D studies, suggested that CCL20 and TNF-alpha are synthesized in response to PAMP stimulation . Taken together, these studies demonstrate that E . coli and selected PAMPs have direct effects on the production of CCL20 and TNF-alpha without affecting cell integrity . Since CCL20 is known to be both chemotactic and antimicrobial, the increase in apical and basolateral release by UEC in response to PAMPs suggests a new mechanism of innate immune protection in the female reproductive tract.

Int J Food Microbiol, 2005 Jan 15, 98(1), 81 - 8
Influence of polysaccharides on oxygen dependent lactate utilization by an amylolytic Lactobacillus plantarum strain; Pintado J et al.; Oxygen-dependent conversion of lactate to acetate by the amylolytic strain Lactobacillus plantarum A6 was studied using MRS-grown cells, transferred to a basic medium with lactate . In the presence of oxygen, lactate was stoechiometricaly converted to acetate . When glucose, maltose or cellobiose was added to the basic medium, no utilisation of lactate was observed . However, when starch or glycogen was added, the conversion of lactate to acetate happened . To verify the possible link of this effect with sugar consumption rate, a glucose-fed batch culture was conducted with a lactic acid consuming culture grown on the basic medium with lactate . Even when glucose was fed at the same low rate as the consumption rate observed for polysaccharides, lactic acid was no more consumed . For the amylolytic strain L . plantarum A6, the transport and use of oligosaccharides resulting from polysaccharides hydrolysis might affect differently the glycolytic flux, with the putative consequence to suppress the metabolic control of glycolysis by glucose . This fact could play an important role in the fermentation of amylaceous foods.

Int J Food Microbiol, 2005 Jan 15, 98(1), 63 - 72
Phenotypic characterization of lactic acid bacteria from sourdoughs for Altamura bread produced in Apulia (Southern Italy); Ricciardi A et al.; In order to study the composition of the lactic acid bacteria (LAB) community of sourdoughs used for the manufacture of Altamura bread, a traditional durum wheat bread produced in Apulia (Southern Italy), 111 strains of LAB were isolated and characterized . The phenotypic characterization of the isolates, carried out using a set of 29 tests, allowed the identification of 15 clusters at the 80% similarity level by hierarchical cluster analysis . Of the isolates, 88% were identified as facultatively heterofermentative LAB (Lactobacillus plantarum, Lb . paracasei, Lb . casei) and 12% as heterofermentative LAB (Lb . brevis, Leuconostoc mesenteroides) . SDS-PAGE profiles of whole cell proteins of 68 strains confirmed the identification . Both the diversity and structure of the lactic microflora for sourdoughs for Altamura bread varied among samples.

Int J Food Microbiol, 2005 Jan 15, 98(1), 53 - 62
A putative glucan synthase gene dps detected in exopolysaccharide-producing Pediococcus damnosus and Oenococcus oeni strains isolated from wine and cider; Walling E et al.; Some lactic acid bacteria can induce viscosity in wine, beer and cider by production of exopolysaccharides (EPS) . A polymerase chain reaction (PCR) assay was previously described for the detection of ropy Pediococcus damnosus strains in wine {J . Appl . Microbiol . 90 (2001) 535} . The primers used in that study, PF5 and PF6, are investigated in addition to new primers which broaden the range of spoiling agents detectable by PCR . Primers PF1 and PF8 allow the amplification of DNA from ropy P . damnosus strains isolated from wine, as was observed with PF5 and PF6 . In addition, PF1 and PF8, unlike PF5 and PF6, are able to generate an amplicon using template DNA from a ropy P . damnosus strain isolated from cider and a ropy Oenococcus oeni strain isolated from champagne . Two different ropy Lactobacillus species were also isolated, but their DNA was not amplified using primers PF1 and PF8 . The new primers PF1 and PF8 were chosen from the sequence of gene dps, a putative glucan synthase gene, found across all the ropy P . damnosus strains isolated, from both wine or cider, and also in a ropy O . oeni strain . To our knowledge, this is the first time that an EPS-producing O . oeni strain is described . Glucan biosynthesis was assessed by agglutination tests done with Streptococcus pneumoniae type 37-specific antibodies, which specifically detect glucan-producing cells . The results show that there is a direct correlation between glucan production and detection of gene dps . Therefore, Dps is considered a candidate for the glucan synthase enzyme responsible for EPS production by ropy strains of P . damnosus and O . oeni.

Antimicrob Agents Chemother, 2005 Jan, 49(1), 170 - 5
Ecological effects of perorally administered pivmecillinam on the normal vaginal microflora; Sullivan A et al.; The knowledge of the effects of antimicrobial agents on the normal vaginal microflora is limited . The objective of the present study was to study the ecological impact of pivmecillinam on the normal vaginal microflora . In 20 healthy women, the estimated day of ovulation was determined during three subsequent menstrual cycles . Microbiological and clinical examinations were performed on the estimated day of ovulation and on day 3 in all cycles and also on day 7 after ovulation in cycles 1 and 2 . Anaerobic and facultative anaerobic gram-positive rods, mainly species of lactobacilli and actinomycetes, dominated the microflora . One woman was colonized on the third day of administration with a resistant Escherichia coli strain, and Candida albicans was detected in one woman on days 3 and 7 in cycle 2 . No other major changes in the normal microflora occurred during the study . Administration of pivmecillinam had a minor ecological impact on the normal vaginal microflora.

Genet Mol Res, 2004 Sep 30, 3(3), 432 - 40
Reduction of non-digestible oligosaccharides in soymilk: application of engineered lactic acid bacteria that produce alpha-galactosidase; LeBlanc JG et al.; Human consumption of soy-derived products has been limited by the presence of non-digestible oligosaccharides (NDO), such as the alpha-galactooligosaccharides raffinose and stachyose . Most mammals, including man, lack pancreatic alpha-galactosidase (alpha-Gal), which is necessary for the hydrolysis of these sugars . However, such NDO can be fermented by gas-producing microorganisms present in the cecum and large intestine, which in turn can induce flatulence and other gastrointestinal disorders in sensitive individuals.The use of microorganisms expressing alpha-Gal is a promising solution to the elimination of NDO before they reach the large intestine . In the present study, lactic acid bacteria engineered to degrade NDO have been constructed and are being used as a tool to evaluate this solution . The alpha-Gal structural genes from Lactobacillus plantarum ATCC8014 (previously characterized in our laboratory) and from guar have been cloned and expressed in Lactococcus lactis . The gene products were directed to different bacterial compartments to optimize their possible applications . The alpha-Gal-producing strains are being evaluated for their efficiency in degrading raffinose and stachyose: i) in soymilk fermentation when used as starters and ii) in situ in the upper gastrointestinal tract when administered to animals orally, as probiotic preparations . The expected outcomes and possible complications of this project are discussed.

Appl Microbiol Biotechnol . 2004 Dec 22; {Epub ahead of print}
Production of Lactobacillus kefir cells for asymmetric synthesis of a 3,5-dihydroxycarboxylate; Pfruender H et al.; An efficient fedbatch process for the production of Lactobacillus kefir DSM 20587 cells was developed . An improvement in space time yield of 270% (3.7 g(DCW) l(-1) day(-1)) and in final enzyme activity of 440% (9.1 U/ml) was achieved on a 150 l scale by controlling the oxygen transfer rate to 7-9 mmol l(-1) h(-1) . The cells exhibited good and highly stereoselective reducing activities against tert-butyl 6-chloro-3,5-dioxohexanoate . tert-Butyl (3R,5S)-6-chloro-dihydroxyhexanoate, a chiral building block for HMG-CoA reductase inhibitor synthesis, was produced with 47.5% yield and >99% ee at C(3)3 and C(5)5 in a simple batch biotransformation process.

Lett Appl Microbiol, 2005, 40(1), 56 - 62
Gene organization and sequences of pediocin AcH/PA-1 production operons in Pediococcus and Lactobacillus plasmids; Miller KW et al.; Abstract k.w . miller, p . ray, t . steinmetz, t . hanekamp and b . ray 2004.Aims: To determine the locations and sequences of pediocin AcH production genes in Pediococcus parvulus ATO77 from vegetables, Lactobacillus plantarum WHE92 from Muenster cheese, and a lactose-fermenting isolate Pediococcus pentosaceus S34 from buffalo milk . Methods and Results: Plasmid curing, Southern blot hybridization, and DNA sequence analysis indicate that pediocin AcH production genes are encoded by highly similar operons in unique plasmids designated pATO77 from P . parvulus ATO77, pS34 from P . pentosaceus S34, and pWHE92 from Lact . plantarum WHE92 . Structure, immunity and secretion system genes are linked together in the operons, and the promoter sequences are the same . The amino acid sequences of the encoded proteins are highly conserved between plasmids . Conclusions: Pediocin AcH production genes are located within a plasmid-borne operon cassette in all lactic acid bacterial strains examined to date . All four genes needed for production are present within a single plasmid in each strain . Significance and Impact of the Study: This is the first demonstration that the expression of a class IIa bacteriocin is directed by a common gene cassette that has been disseminated to unique plasmids in different genera of lactic acid bacteria . These plasmids should be useful for expressing pediocin AcH in Pediococcus and Lactobacillus strains used in food production.

J Agric Food Chem, 2004 Dec 29, 52(26), 7752 - 8
High-performance liquid chromatographic determination of naturally occurring folates during tempe preparation; Ginting E et al.; A trienzyme treatment (protease, alpha-amylase, and human plasma conjugase), followed by purification using SPE with SAX cartridges and reversed-phase HPLC with UV-PDA detection, was performed for determination of the distribution of various folate forms and content at various stages of tempe preparation . The major folate form in soybean identified was 5-formyl tetrahydrofolate (5-CHO-H4folate), followed by 10-formyl tetrahydrofolate (10-CHO-PGA), and 5-methyl tetrahydrofolate (5-CH3-H4folate), whereas folic acid was not detected and tetrahydrofolic acid (H4folate) was not detectable . The most predominant form in tempe was also 5-CHO-H4folate, followed by 10-CHO-PGA, whereas the quantities of 5-CH3-H4folate and folic acid were negligible . Quantities and retention of folate significantly decreased during the first boiling, dehulling, soaking, and second boiling procedures, yielding folate retention of 32% . A remarkable increase in folate content was found after fermentation, 5.2-fold higher than that of the boiled soybean . This may be due to de novo formation of folate by Rhizopus oligosporus, the principal mold in tempe fermentation . HPLC results were approximately 38-55% lower than the values obtained from the microbiological assay using Lactobacillus casei.

J Appl Microbiol, 2005, 98(1), 229 - 37
Characterization of an acid phosphatase from Lactobacillus pentosus: regulation and biochemical properties; Palacios MC et al.; Abstract m.c . palacios, m . haros, c.m . rosell and y . sanz . 2004.Aims: To screen for phosphatase and phytase activities in Lactobacillus isolated from diverse ecosystems and to determine the biochemical properties and the factors that regulate the synthesis of the enzyme responsible for these activities in the selected strain, Lactobacillus pentosus CECT 4023 . Methods and Results: These activities were determined spectrophotometrically by using p-nitrophenyl phosphate and sodium phytate as substrates . They were maximal at the onset of the stationary phase of growth and repressed in the presence of high glucose concentration and inorganic phosphate . The enzyme responsible for these activities was an acid phosphatase (E.C.3.1.3.2.), with a molecular mass of 69 kDa . The activity was optimum at pH 5.0 and 50 degrees C . It hydrolysed mono-phosphorylated substrates and phytate, albeit at lower rates . It was inhibited by iodoacetic acid, phenyl-methylsulphonyl fluoride, di-sodium pyrophosphate and Ca(+2) while activated by Co(+2) and low concentrations of l-ascorbic acid and EDTA . Conclusions: Lactobacillus pentosus CECT 4023 produces a nonspecific acid phosphatase that hydrolyses a number of mono-phosphorylated substrates and phytate . Significance and Impact of the Study: The results suggest that the phosphatase from L . pentosus CECT 4023 could partly contribute to reduce the phosphorylation degree of phytate and its derivatives and, thereby, their anti-nutrient properties during fermentation processes.

J Appl Microbiol, 2005, 98(1), 84 - 95
Cell wall modifications during osmotic stress in Lactobacillus casei; Piuri M et al.; Abstract m . piuri, c . sanchez-rivas and s.m . ruzal . 2004.Aims: To study the modification of the cell wall of Lactobacillus casei ATCC 393 grown in high salt conditions . Methods and Results: Differences in the overall structure of cell wall between growth in high salt (MRS + 1 mol l(-1) NaCl; N condition) and control (MRS; C condition) conditions were determined by transmission electronic microscopy and analytical procedures . Lactobacillus casei cells grown in N condition were significantly larger than cells grown under unstressed C condition . Increased sensitivity to mutanolysin and antibiotics with target in the cell wall was observed in N condition . Purified cell wall also showed the increased sensitivity to lysis by mutanolysin . Analysis of peptidoglycan (PG) from stressed cells showed that modification was at the structural level in accordance with a decreased PG cross-link involving penicillin-binding proteins (PBP) . Nine PBP were first described in this species and these proteins were expressed in low percentages or presented a modified pattern of saturation with penicillin G (Pen G) during growth in high salt . Three of the essential PBP were fully saturated in N condition at lower Pen G concentrations than in C condition, suggesting differences in functionality in vivo . Conclusions: The results show that growth in high salt modified the structural properties of the cell wall . Significance and Impact of Study: Advances in understanding the adaptation to high osmolarity, in particular those involving sensitivity to lysis of lactic acid bacteria.

J Appl Microbiol, 2005, 98(1), 64 - 72
Analysis of promoter sequences from Lactobacillus helveticus CNRZ32 and their activity in other lactic acid bacteria; Chen YS et al.; Abstract y.-s . chen and j.l . steele . 2004.Aims: To clone and analyse seven putative promoter fragments (pepC, pepN, pepX, pepO, pepE, pepO2, hsp17) from Lactobacillus helveticus CNRZ32 for their expression in Lact . helveticus CNRZ32, Lact . casei ATCC334 and Lactococcus lactis MG1363 . Methods and Results: Promoter fragments were fused to the promoter-less beta-glucuronidase (gusA) gene on pNZ272(RBS-) (ATG-) . The resulting constructs were evaluated for their ability to drive the expression of active GusA with 0.5 mmol l(-1) 5-bromo-4-chloro-3-indolyl-beta-d-glucuronide . All promoters except P(pepN)::gusA were active in the examined strains . Northern hybridization was performed to examine the promoter strength . Sequence analysis of these promoters identified well conserved putative ribosomal binding and putative -10 hexamers sites . Conclusions: Seven promoter fragments from Lact . helveticus CNRZ32 were recognized in the lactic acid bacteria, Lact . casei ATCC334 and L . lactis MG1363, as well as in Escherichia coli . P(pepN)::gusA could not be maintained in the strains examined because of toxicity associated with heterologous protein over-expression driven by P(pepN) . Significance and Impact of the Study: This study revealed that desirable levels of heterologous food-grade protein production in GRAS organisms can be obtained with the application of natural promoter fragments from closely related organisms.

J Appl Microbiol, 2005, 98(1), 56 - 63
Lyophilized preparations of bacteriocinogenic Lactobacillus curvatus and Lactococcus lactis subsp . lactis as potential protective adjuncts to control Listeria monocytogenes in dry-fermented sausages; Benkerroum N et al.; Abstract n . benkerroum, a . Daoudi, t . hamraoui, h . Ghalfi, c . thiry, m . duroy, p . evrart, d . roblain and p . thonart . 2004.Aim: Study of the effectiveness of in situ bacteriocin production by lactic acid bacteria (LAB) to control Listeria monocytogenes in dry-fermented sausages . Methods and Results: Two bacteriocin-producing strains: Lactococcus lactis subsp . lactis LMG21206 and Lactobacillus curvatus LBPE were grown in a pilot scale fermentor and lyophilized to be directly used in dry sausage fermentation . A commercial starter culture (Bel'meat(TM) SL-25) not inhibitory to L . monocytogenes (Bac(-) starter) was mixed (1 : 1) with each of the two lyophilized bacteriocin-producing strains to obtain starters active against the pathogen (Bac(+) starter) . Anti-Listeria effectiveness of the Bac(+) starters was studied in dry-fermented sausages . The meat batter was experimentally contaminated with a mixture of four different strains of L . monocytogenes (10(2)-10(3) CFU g(-1)) . The results showed that L . monocytogenes did not grow in any of the contaminated batches, but no significant decrease (P > 0.05) was observed either in the positive control (no added starter culture) or in samples fermented with the Bac(-) starter culture during the fermentation period and up to 15 days of drying . When the Bac(+) starter contained Lb . curvatus LBPE, cell counts of L . monocytogenes decreased to below the detectable limit (<10 CFU g(-1)) after 4 h of fermentation and no survivors could be recovered by enrichment beyond day 8 of drying . When the Bac(+) starter culture containing Lc . lactis LMG21206 was used, a decrease in Listeria counts to below the detectable limit was achieved after 15 days of drying . Conclusions: The bacteriocin-producing strains studied may be used as adjunct cultures for sausage fermentations to control the occurrence and survival of L . monocytogenes . Significance and Impact of the Study: Addition of the Bac(+) strains, especially the Lb . curvatus strain would provide an additional hurdle to enhance the control of L . monocytogenes in fermented meat products.

J Appl Microbiol, 2005, 98(1), 14 - 23
Succinate production and citrate catabolism by Cheddar cheese nonstarter lactobacilli; Dudley EG et al.; Abstract e.g . dudley and j.l . steele . 2004.Aims: To identify strains of Cheddar cheese nonstarter lactobacilli that synthesize succinate from common precursors and characterize the biochemical pathways utilized . Methods and Results: Whole cell incubations of Lactobacillus plantarum, Lactobacillus casei, Lactobacillus zeae and Lactobacillus rhamnosus, were used to identify strains that accumulated succinate from citrate, l-lactate, aspartic acid or isocitrate . In vivo(13)C-nuclear magnetic resonance spectroscopy ((13)C-NMR) identified the biochemical pathway involved at pH 7.0, and under conditions more representative of the cheese ripening environment (pH 5.1/4% NaCl/13 degrees C) . Enzyme assays on cell-free extracts were used to support the pathway suggested by (13)C-NMR . Conclusions: The Lact . plantarum strains studied synthesize succinate from citrate by the reductive tricarboxylic acid (TCA) cycle at either pH 7.0 or pH 5.1/4% NaCl/13 degrees C . Lactobacillus casei, Lact . zeae and Lact . rhamnosus strains lack one or more enzymatic activities present in this pathway, and do not accumulate succinate from any of the four precursors studied . Significance and Impact of the Study: The addition of Lact . plantarum strains to milk during cheese manufacture may increase the accumulation of the flavour enhancer succinate.

Ann Allergy Asthma Immunol, 2004 Dec, 93(6), 601 - 5
Rice-induced enterocolitis in an infant: TH1/TH2 cellular hypersensitivity and absent IgE reactivity; Gray HC et al.; BACKGROUND: Although food allergy is common in children, rice allergy is unusual in Western cultures . OBJECTIVE: To report a case of T-cell-mediated rice intolerance in an 11-month-old girl . METHODS: To evaluate the intolerance to rice in this patient, a graded rice food challenge was performed . To examine the immunologic reactivity to rice, in vitro lymphoproliferative responses and cytokine synthesis of rice-stimulated peripheral blood lymphocytes (PBLs) was performed . Subsequently, skin patch testing to rice and other foods was performed . RESULTS: Allergy skin prick test results were negative for rice and positive for egg, milk, and soy . Specific IgE antibodies to rice, egg, peanut, wheat, walnut, codfish, milk, soybean, corn, shrimp, scallops, and clams were undetectable . Results of a single-blind rice food challenge were positive, manifested by emesis that persisted for more than an hour and required intravenous hydration . In vitro lymphoproliferation by the patient's PBLs to rice stimulation was positive . In addition, cytokine synthesis of interferon-gamma, interleukin 10 (IL-10), tumor necrosis factor a, and IL-5 by the patient's rice-stimulated PBLs was elevated, indicating a TH1/TH2 cell response to rice . Endoscopy revealed normal esophageal, gastric, and duodenal mucosa; a biopsy specimen revealed mild esophagitis . Duodenal explant T cells were initially established by stimulation with rice and IL-2 . After a 2-day rest, the lymphocytes were restimulated with rice for 7 days and revealed increased interferon-gamma and IL-5 synthesis . Twenty billion colony forming units of Lactobacillus GG were added to the patient's diet twice daily . After 6 weeks, rice rechallenge resulted in emesis within 1 hour . Results of patch testing were positive to rice, wheat, and barley but negative to soy, which the patient tolerated on food challenge . CONCLUSIONS: Although this patient did not demonstrate IgE antibody to rice, TH1/TH2 cell-mediated responses to rice were detected, and the patient experienced significant morbidity . Patch testing for gastrointestinal food allergies may be useful when the food specific IgE antibody is negative . Probiotic therapy in this patient did not ameliorate her sensitivity to rice, and food elimination remains the only reliable treatment for TH1/TH2-mediated food hypersensitivity.

J Dairy Res, 2004 Nov, 71(4), 461 - 70
Amino acid transamination in permeabilised cells of Lactobacillus helveticus, Lb . paracasei and Lb . danicus; Thage BV et al.; Aminotransferase (AT) activity against 18 amino acids was studied in ten strains of three species of Lactobacillus . A method for permeabilisation of cells was developed using toluene and ethanol combined with mechanical treatment . It was found that the AT activities in the washed permeabilised cells (W-PC) corresponded well to that in cell-free extracts (CFE) . The AT specificity pattern was species as well as strain dependant . Strains of Lb . helveticus had high specificity for aromatic amino acids (ArAA) and lower activity against branched-chain amino acids (BcAA) and Asp, while strains of Lb . paracasei subsp . paracasei degraded BcAA and Asp, but had a lower and variable specificity against ArAA . One of the Lb . paracasei strains was characterised by having very high AT activity against all three BcAA (Ile, Leu, Val) compared with any of the other Lb . paracasei strains tested . Strains of Lb . danicus, which is a newly discovered Lactobacillus species isolated from cheese, had up to about 20 times higher AT activity against Leu than Lb . paracasei and Lb . helveticus . The permeabilised cells of Lb . danicus had also considerably higher AT activity against ArAA than Lb . paracasei and Lb . helveticus strains, and also higher AT activity against Asp . All Lactobacillus strains tested had AT activity against Met, but at a much lower rate than against other amino acids . Results of this study also demonstrated a chemical reaction between alpha-ketoglutaric acid and Asp that was catalysed by pyridoxal-5-phosphate without any AT present.

J Dairy Res, 2004 Nov, 71(4), 451 - 60
In vitro screening of potential probiotic activities of selected lactobacilli isolated from unpasteurized milk products for incorporation into soft cheese; Coeuret V et al.; The aim was to select potentially probiotic lactobacilli from 88 strains isolated from unpasteurized milk and cheese products, and to incorporate these bacteria in a viable state into a soft cheese, without changing its quality . The survival of these bacteria was assessed in acidic and bile conditions, after freezing at -80 degrees C . Four strains from unpasteurized Camembert--two Lactobacillus plantarum strains and two Lb . paracasei/casei strains--were identified and typed by PCR and PFGE and were found to display potentially probiotic characteristics in addition to resistance to low pH and bile . These characteristics were resistance to lysozyme, adhesion to CACO-2 cells, antimicrobial effects against common foodborne pathogens (Listeria monocytogenes, Staphylococcus aureus, Salmonella spp., Escherichia coli, innocuity following the ingestion of high doses by mice and appropriate antibiotic susceptibility profiles . The potential of Lb . plantarum strain UCMA 3037 for incorporation into a soft cheese (Pont-l'Eveque registered designation of origin (RDO)) was investigated . This strain grew well and survived in sufficient numbers (more than 10(7) cfu/g throughout the shelf-life of the product) in the cheese . This strain did not change the quality score of the product until the best before date (75 days after manufacture) . Thus, unpasteurized Camembert is a natural source of potentially probiotic lactobacilli, which could be used as an additive in the development of potentially probiotic soft cheeses . Further work is required to demonstrate the persistence and efficacy of these strains in the human host upon ingestion.

Biotechnol Lett, 2004 Nov, 26(22), 1739 - 44
Improved synthesis of chiral alcohols with Escherichia coli cells co-expressing pyridine nucleotide transhydrogenase, NADP(+)-dependent alcohol dehydrogenase and NAD(+)-dependent formate dehydrogenase; Weckbecker A et al.; Recombinant pyridine nucleotide transhydrogenase (PNT) from Escherichia coli has been used to regenerate NAD(+) and NADPH . The pnta and pntb genes encoding for the alpha- and beta-subunits were cloned and co-expressed with NADP(+)-dependent alcohol dehydrogenase (ADH) from Lactobacillus kefir and NAD(+)-dependent formate dehydrogenase (FDH) from Candida boidinii . Using this whole-cell biocatalyst, efficient conversion of prochiral ketones to chiral alcohols was achieved: 66% acetophenone was reduced to ( R )-phenylethanol over 12 h, whereas only 19% ( R )-phenylethanol was formed under the same conditions with cells containing ADH and FDH genes but without PNT genes . Cells that were permeabilized with toluene showed ketone reduction only if both cofactors were present.

Biotechnol Lett, 2004 Oct, 26(20), 1613 - 6
Fermentative production of DL-lactic acid from amylase-treated rice and wheat brans hydrolyzate by a novel lactic acid bacterium, Lactobacillus sp; Yun JS et al.; Rice and wheat brans, without additional nutrients and hydrolyzed by alpha-amylase and amyloglucosidase, were fermented to DL-lactic acid using a newly isolated strain of Lactobacillus sp . RKY2 . In batch fermentations at 36 degrees C and pH 6, the amount of lactic acid in fermentation broth reached 129 g l(-1) by supplementation of rice bran with whole rice flour . The maximum productivity was 3.1 g lactic acid l(-1) h(-1) in rice bran medium supplemented with whole rice flour or whole wheat flour.

Ann N Y Acad Sci, 2004 Nov, 1026, 284 - 8
Immunization of Bovines with Concealed Antigens from Haematobia irritans; Bautista CR et al.; To evaluate an immunization procedure using antigens from Haematobia irritans intestine (AgHiI), four bovines (group I) were inoculated with AgHiI mixed with Freund's incomplete adjuvant containing Lactobacillus casei, three bovines (group II) received AgHiI, and three bovines (group III) received saline solution . At day 35, blood was collected from each animal to feed H . irritans flies . There was no difference in the fly mortality observed in the three groups . The percentage of reduction of eggs oviposited by each female in 8 days (%RE), as compared with group III, was 29.45 for group I and 11.02 for group II . Antibody levels (AbL) to AgHiI were higher in group I than in groups II and III . A high correlation between %RE and AbL was observed.

Stomatologiia (Mosk), 2004, 83(6), 6 - 8
{Development and use of periodontal dressing of collagen and Lactobacillus casei 37 cell suspension in combined treatment of periodontal disease of inflammatory origin (results of microbiologic study)}; Effect of yeast inoculation rate on the metabolism of contaminating lactobacilli during fermentation of corn mash; Alltech Biotechnology Center, 3031 Catnip Hill Pike, Nicholasville, KY, 40356, USA, nnarendranath@alltech.comTwo separate 4 (bacterial concentrations)x6 (yeast concentrations) full factorial experiments were conducted in an attempt to identify a novel approach to minimize the effects caused by bacterial contamination during industrial production of ethanol from corn . Lactobacillus plantarum and Lactobacillus paracasei, commonly occurring bacterial contaminants in ethanol plants, were used in separate fermentation experiments conducted in duplicate using an industrial strain of Saccharomyces cerevisiae, Allyeast Superstart . Bacterial concentrations were 0, 1x10(6), 1x10(7) and 1x10(8) cells/ml mash . Yeast concentrations were 0, 1x10(6), 1x10(7), 2x10(7), 3x10(7), and 4x10(7) cells/ml mash . An increased yeast inoculation rate of 3x10(7) cells/ml resulted in a greater than 80% decrease (P<0.001) and a greater than 55% decrease (P<0.001) in lactic acid production by L . plantarum and L . paracasei, respectively, when mash was infected with 1x10(8) lactobacilli/ml . No differences (P>0.25) were observed in the final ethanol concentration produced by yeast at any of the inoculation rates studied, in the absence of lactobacilli . However, when the mash was infected with 1x10(7) or 1x10(8) lactobacilli/ml, a reduction of 0.7-0.9% v/v (P<0.005) and a reduction of 0.4-0.6% v/v (P<0.005) in the final ethanol produced was observed in mashes inoculated with 1x10(6) and 1x10(7) yeast cells/ml, respectively . At higher yeast inoculation rates of 3x10(7) or 4x10(7) cells/ml, no differences (P>0.35) were observed in the final ethanol produced even when the mash was infected with 1x10(8) lactobacilli/ml . The increase in ethanol corresponded to the reduction in lactic acid production by lactobacilli . This suggests that using an inoculation rate of 3x10(7) yeast cells/ml reduces the growth and metabolism of contaminating lactic bacteria significantly, which results in reduced lactic acid production and a concomitant increase in ethanol production by yeast.

Eur J Clin Microbiol Infect Dis . 2004 Dec 14; {Epub ahead of print}
Pathogenic relevance of Lactobacillus: a retrospective review of over 200 cases; Cannon JP et al.; Given that Lactobacillus has been reported to be the causative pathogen in many types of infection despite debate regarding the organism's clinical significance, a literature review was conducted to investigate the treatments and outcomes of Lactobacillus infections reported to date . In this article, the characteristics of over 200 reported cases of Lactobacillus-associated infections are summarized . Lactobacillus was found to be frequently associated with endocarditis and bacteremia . Lactobacillus was also associated with a variety of other infections including, but not limited to, peritonitis, abscesses, and meningitis . The species casei and rhamnosus were the most common . The isolates tended to be most sensitive to erythromycin and clindamycin and most resistant to vancomycin . The species that was most sensitive to vancomycin was acidophilus . The overall mortality rate was nearly 30% . There was a significant association between mortality and polymicrobial infection (P=0.004) . In the subset of patients with bacteremia, increased mortality was associated with inadequate treatment (P=0.001) and polymicrobial bacteremia (P=0.044).

Appl Microbiol Biotechnol, 2005 Jan, 66(4), 414 - 21 Epub 2004 Jul 23.
Quantification by real-time PCR of Lactococcus lactis subsp . cremoris in milk fermented by a mixed culture; Grattepanche F et al.; During cheese making, interactions between different strains of lactic acid bacteria play an important role . However, few methods are available to specifically determine each bacterial population in mixed cultures, in particular for strains of the same species . The aim of this study was to develop a real-time PCR quantification method to monitor the population of Lactococcus cremoris ATCC 19257 in mixed culture with Lactobacillus rhamnosus RW-9595M and the bacteriocin-producing microorganism Lc . diacetylactis UL719 . The specificity of the two primers 68FCa33 and 16SR308 used to amplify a 240-bp fragment of DNA from Lc . cremoris was demonstrated by conventional PCR . Using these primers for real-time PCR, the detection limit was 2 cfu/reaction or 200 cfu of Lc . cremoris ATCC 19257 per millilitre of mixed culture in milk . In pure culture batch fermentation, good correlation was obtained between real-time PCR and the conventional plating method for monitoring Lc . cremoris growth . In mixed culture batch fermentation, Lb . rhamnosus and Lc . cremoris decreased due to nisin Z production by Lc . diacetylactis . The decrease of the Lc . cremoris cell population detected by real-time PCR was not possible to observe by the plate count method in the presence of a Lc . diacetylactis population that was 1 log higher.

Biotechnol Bioeng, 2005 Jan 20, 89(2), 129 - 37
Fractionation of cell mixtures using acoustic and laminar flow fields; Kumar M et al.; A fractionation method applicable to different populations of cells in a suspension is reported . The separation was accomplished by subjecting the suspension to a resonant ultrasonic field and a laminar flow field propagating in orthogonal directions within a thin, rectangular chamber . Steady, laminar flow transports the cell suspension along the chamber, while the ultrasonic field causes the suspended cells to migrate to the mid-plane of the chamber at rates related to their size and physical properties . A thin flow splitter positioned near the outlet divides the effluent cell suspension into two product streams, thereby allowing cells that respond faster to the acoustic field to be separated from those cells that respond more slowly . Modeling of the trajectories of individual cells through the chamber shows that by altering the strength of the flow relative to that of the acoustic field, the desired fractionation can be controlled . Proof-of-concept experiments were performed using hybridoma cells and Lactobacillus rhamnosus cells . The two populations of cells could be effectively separated using this technique, resulting in hybridoma/Lactobacillus ratios in the left and right product streams, normalized to the feed ratio, of 6.9 +/- 1.8 and 0.39 +/- 0.01 (vol/vol), respectively . The acoustic method is fast, efficient, and could be operated continuously with a high degree of selectivity and yield and with low power consumption . (c) 2004 Wiley Periodicals, Inc.

J Infect Dis, 2005 Jan 1, 191(1), 25 - 32 Epub 2004 Dec 02.
Female genital-tract HIV load correlates inversely with Lactobacillus species but positively with bacterial vaginosis and Mycoplasma hominis; Sha BE et al.; BACKGROUND: Bacterial vaginosis (BV) is associated with human immunodeficiency virus (HIV) acquisition . We examined the association between BV and BV-associated bacteria and expression of HIV in the female genital tract . METHODS: HIV RNA, lactobacilli, Gardnerella vaginalis, and Mycoplasma hominis in cervicovaginal lavage (CVL) samples were quantified by polymerase chain reaction (PCR) . Gynecologic evaluation included Nugent score assessment, Amsel criteria assessment, detection of other genital-tract infections, and dysplasia grading . CD4 cell count, plasma HIV RNA level, and antiretroviral history were obtained . RESULTS: A total of 203 CVL samples from women with Nugent scores of 7-10 (BV group) and 203 samples from women with Nugent scores of 0-3 (no-BV group) were matched by plasma HIV RNA level and analyzed . After controlling for plasma HIV RNA level and Nugent score in univariate analyses, we found that G . vaginalis and M . hominis bacterial counts, Candida vaginitis, and herpes simplex virus (HSV) were positively associated with CVL HIV RNA levels . In multivariate analysis, only lactobacilli bacterial counts (P=.006; inverse association), M . hominis bacterial counts (P=.0001; positive association), Candida vaginitis (P=.007), and HSV (P=.03) were significantly associated with CVL HIV RNA levels . CONCLUSION: Bacteria associated with BV increase genital-tract HIV RNA levels . Quantitative bacterial counts for lactobacilli and M . hominis are better correlates of CVL HIV RNA than are Nugent score or Amsel criteria . Since plasma virus and CD4 cell levels did not differ between the BV and no-BV groups, these data suggest that the bacterial flora associated with BV influence genital-tract HIV shedding.

Caries Res, 2005 Jan-Feb, 39(1), 41 - 7
Caries prevalence measured with QLF after treatment with fixed orthodontic appliances: influencing factors; Boersma JG et al.; Caries prevalence on the buccal surfaces of teeth in orthodontic patients was determined with QLF and visual examination immediately after removal of fixed appliances . The number of lesions found by QLF far outnumbered that found by visual examination, but the distribution pattern was similar . 97% of all subjects and on average 30% of the buccal surfaces in a person were affected . On average, in males 40% of surfaces and in females 22% showed white spots (p < 0.01) . Caries prevalence was lower (p < 0.01) in incisors and cuspids than in molars and premolars . A positive correlation with caries prevalence was found for the bleeding scores 6 weeks after debonding and lactobacillus counts before debonding . Mutans streptococci counts, age, treatment duration, socioeconomic status and dietary habits showed no correlation with caries prevalence .

J Dairy Sci, 2005 Jan, 88(1), 55 - 66
Acid and bile tolerance and cholesterol removal ability of lactobacilli strains; Liong MT et al.; Eleven strains of lactobacilli were studied for their acid and bile tolerance . Possible mechanisms of cholesterol removal by strains of lactobacilli were examined . Cholesterol assimilation as determined by the difference in cholesterol content in the medium before and after the incubation period showed that all lactobacilli strains were able to assimilate cholesterol at varying levels ranging from 12.03 to 32.25 mug/mL . Cholesterol removal was associated with growth of cultures . Binding of cholesterol to lactobacilli cells was determined using growing, heat-killed, and resting cells in phosphate buffer . Cholesterol removed by dead and resting cells ranged from 0.79 to 3.82 mg/g of dry weight compared with growing cells, which ranged from 4.53 to 16.03 mg/g of dry weight . Fatty acid methyl esters, as quantified using gas chromatography, showed changes in lipid profiles in cells grown in the presence of cholesterol compared with those grown without cholesterol . Fatty acid profiles, especially of hexadecanoic, octadecanoic, total saturated, and unsaturated acids suggested that cholesterol from the medium was incorporated into the cellular membrane . These findings suggest that strains of lactobacilli could remove cholesterol via various mechanisms and may be promising candidates for use as a dietary adjunct to lower serum cholesterol in vivo.

J Dairy Sci, 2005 Jan, 88(1), 21 - 9
Fermentation pH and Temperature Influence the Cryotolerance of Lactobacillus acidophilus RD758; Wang Y et al.; The effects of 3 fermentation temperatures (30, 37, and 42 degrees C) and 3 fermentation pH (4.5, 5, and 6) on the cryotolerance of Lactobacillus acidophilus RD758 were studied in relation to their fatty acid composition . Cryotolerance was defined as the ability of the cells to recover their acidification activity after freezing and frozen storage at -20 degrees C . Better cryotolerance was obtained in cells grown at 30 degrees C or at pH 5; these cells showed no loss in acidification activity during freezing and a low rate of loss in acidification activity during frozen storage . On the other hand, cells grown at 42 degrees C or at pH 4.5 displayed poor cryotolerance . The membrane fatty acid composition was analyzed and related to the cryotolerance using principal component analysis . The improved cryotolerance observed during the freezing step was associated with a high ratio of unsaturated to saturated fatty acids, a low C18:0 content, and high C16:0 and cyclic C19:0 relative concentrations . High resistance during frozen storage was related to a high cycC19:0 concentration . Finally, the low cryotolerance observed after fermentation at pH 4.5 was explained by a low C18:2 content.

Am J Clin Nutr, 2004 Dec, 80(6), 1618 - 25
Lactobacillus rhamnosus induces peripheral hyporesponsiveness in stimulated CD4+ T cells via modulation of dendritic cell function; Braat H et al.; BACKGROUND: Although it is widely recognized that the intake of so-called probiotic microorganisms is beneficial in chronic mucosal inflammation and topical allergic disease, the immunologic details explaining how such bacteria can exert these effects remain obscure . OBJECTIVE: We determined whether Lactobacillus rhamnosus can modulate T cell responses in vitro and in vivo . DESIGN: In vitro, human monocyte-derived dendritic cells (DCs) matured in the presence of L . rhamnosus were used to instruct naive CD4+ T cells; subsequently, the T cell response was assessed with the use of CD3/CD28 and interleukin (IL) 2 . Cytokine production by ex vivo-stimulated naive cells and memory T cells was measured before and after oral supplementation with L . rhamnosus in 6 healthy volunteers and 6 patients with Crohn disease . RESULTS: A decreased T cell proliferation and cytokine production, especially of IL-2, IL-4, and IL-10, was observed in CD3/CD28-stimulated T cells derived from L . rhamnosus-matured DCs . This T cell hyporesponsiveness was associated with enhanced DC-T cell interaction and normal responsiveness of T cells for IL-2 . In vivo oral supplementation of L . rhamnosus for 2 wk induced a similar T cell hyporesponsiveness, including impaired ex vivo T helper subsets 1 and 2 responses without up-regulation of immunoregulatory cytokines in cohorts of both healthy volunteers and patients with Crohn disease . CONCLUSIONS: We propose that L . rhamnosus modulates DC function to induce a novel form of T cell hyporesponsiveness; this mechanism might be an explanation for the observed beneficial effects of probiotic treatment in clinical disease.

Int J Food Microbiol, 2005 Jan 1, 97(3), 307 - 15
Water activity affects heat resistance of microorganisms in food powders; Laroche C et al.; To study the factors and mechanisms involved in microorganisms' death or resistance to temperature in low-water-activity environments, a previous work dealt with the viability of dried microorganisms immobilized in thin-layer on glass beads . This work is intended to check the efficiency of a rapid heating-cooling treatment to destroy microorganisms that were dried after mixing with wheat flour or skim milk . The thermoresistance of the yeast Saccharomyces cerevisiae and the bacterium Lactobacillus plantarum were studied . Heat stress was applied at two temperatures (150 or 200 degrees C) for treatments of one of four durations (5, 10, 20, or 30 s) and at seven levels of initial water activity (a(w)) in the range 0.10 to 0.70 . This new treatment achieved a microbial destruction of eight log reductions . A specific initial water activity was defined for each strain at which it was most resistant to heat treatments . On wheat flour, this initial a(w) value was in the range 0.30-0.50, with maximal viability value at a(w)=0.35 for L . plantarum, whatever the temperature studied, and 0.40 for S . cerevisiae . For skim milk, a variation in microbial viability was observed, with optimal resistance in the range 0.30-0.50 for S . cerevisiae and 0.20-0.50 for L . plantarum, with minimal destruction at a(w)=0.30 whatever the heating temperature is.

Int J Food Microbiol, 2005 Jan 1, 97(3), 285 - 96
Characterization and identification of lactic acid bacteria in "morcilla de Burgos"; Santos EM et al.; A total of 176 lactic acid bacteria (LAB) isolated from a typical Spanish blood sausage called "morcilla de Burgos" were identified by means of phenotypic characteristics and 16S rDNA RFLP (ribotyping) . LAB were isolated from "morcilla" of different producers and in different storage periods, which includes unpackaged, vacuum and modified atmosphere packaged "morcilla" and vacuum packed and pasteurised "morcilla" . The knowledge of specific spoilage bacteria of "morcilla de Burgos" will be useful to design new preservation methods to extend the shelf-life of this product . Identification made according to phenotypic and biochemical characteristics shows the majority of the isolates were heterofermentative LAB (93.2%) and eight different bacterial groups could be distinguished (A-G) . Weisella viridescens was the main species detected (42%) . In addition, Leuconostoc spp . (23.9%), Weissella confusa (11.4%) and Lactobacillus fructosus (5.7%) species were found . Few strains were phenotypically missidentified as Lactobacillus sanfrancisco, Pediococcus spp., Lactobacillus sakei/curvatus and Carnobacterium spp . and 11 strains remained unknown . Most of the leuconostocs were identified as Leuconostoc mesenteroides and Leuconostoc carnosum species . Ribotyping shows a quite good correlation with phenotypic methods, although it has been possible to identify 15 different clusters . W . viridescens and leuconostocs were also the predominant LAB . Strains identified as W . confusa by phenotypic characteristics were resolved in W . confusa and Weissella cibaria by ribotyping . Neither Carnobacterium piscicola nor Lb . sanfrancisco were identified by means of genotypic method . All Lb . fructosus strains and some more included in different phenotypic groups (17 strains in total) could not be associated with any reference strain (cluster VII).

J Fam Pract . 2004 Dec;53(12):952.
Lactobacillus does not prevent post-antibiotic vaginitis; Solution structure and thermolysis of Cobeta-5'-deoxyadenosylimidazolylcobamide et al.; Department of Chemistry and Biochemistry, Ohio University, Athens, Ohio 45701, USA . brownk3@ohiou.edu

The solution structure of Cobeta-5'-deoxyadenosylimidazolylcobamide, Ado(Im)Cbl, the coenzyme B(12) analogue in which the axial 5,6-dimethylbenzimidazole (Bzm) ligand is replaced by imidazole, has been determined by NMR-restrained molecular modeling . A two-state model, in which a conformation with the adenosyl moiety over the southern quadrant of the corrin and a conformation with the adenosyl ligand over the eastern quadrant of the corrin are both populated at room temperature, was required by the nOe data . A rotation profile and molecular dynamics simulations suggest that the eastern conformation is the more stable, in contrast to AdoCbl itself in which the southern conformation is preferred . Consensus structures of the two conformers show that the axial Co-N bond is slightly shorter and the corrin ring is less folded in Ado(Im)Cbl than in AdoCbl . A study of the thermolysis of Ado(Im)Cbl in aqueous solution (50-125 degrees C) revealed competing homolytic and heterolytic pathways as for AdoCbl but with heterolysis being 9-fold faster and homolysis being 3-fold slower at 100 degrees C than for AdoCbl . Determination of the pK(a)'s for the Ado(Im)Cbl base-on/base-off reaction and for the detached imidazole ribonucleoside as a function of temperature permitted correction of the homolysis and heterolysis rate constants for the temperature-dependent presence of the base-off species of Ado(Im)Cbl . Activation analysis of the resulting rate constants for the base-on species show that the entropy of activation for Ado(Im)Cbl homolysis (13.7 +/- 0.9 cal mol(-1) K(-1)) is identical with that of AdoCbl (13.5 +/- 0.7 cal mol(-1) K(-1)) but that the enthalpy of activation (34.8 kcal mol(-1)) is 1.0 +/- 0.4 kcal mol(-1) larger . The opposite effect is seen for heterolysis, where the enthalpies of activation are identical but the entropy of activation is 5 +/- 1 cal mol(-1) K(-1) less negative for Ado(Im)Cbl . Extrapolation to 37 degrees C provides a rate constant for Ado(Im)Cbl homolysis of 2.1 x 10(-9) s(-1), 4.3-fold smaller than for AdoCbl . Combined with earlier results for the enzyme-induced homolysis of Ado(Im)Cbl by the ribonucleoside triphosphate reductase from Lactobacillus leichmannii, the catalytic efficiency of the enzyme for homolysis of Ado(Im)Cbl at 37 degrees C can be calculated to be 4.0 x 10(8), 3.8-fold, or 0.8 kcal mol(-1), smaller than for AdoCbl . Thus, the bulky Bzm ligand makes at best a <1 kcal mol(-1) contribution to the enzymatic activation of coenzyme B(12).

Clin Infect Dis, 2004 Dec 15, 39(12), 1858 - 60 Epub 2004 Dec 15.
Phenotypic differences between commercial Lactobacillus rhamnosus GG and L . rhamnosus strains recovered from blood; Ouwehand AC et al.; The isolation of clinical isolates that are indistinguishable, by molecular methods, from the probiotic strain of Lactobacillus rhamnosus GG has been reported . We compared the virulence potential of these clinical isolates with that of the probiotic strain . It was observed that all isolates differed significantly in 1 or more phenotypic properties.

Eur J Nutr . 2004 Dec 1; {Epub ahead of print}
Probiotic bacteria stimulate virus-specific neutralizing antibodies following a booster polio vaccination; de Vrese M et al.; BACKGROUND . Orally ingested probiotic bacteria may modulate the immune response and increase antibody titers against enteric infections by bacteria or viruses . Even though positive effects of probiotics on respiratory tract infections have been reported, overall only few studies have examined effects on virus infections concerning organs other than the gastrointestinal tract . AIM OF THE STUDY . It was the aim of the study to investigate whether and how probiotics affect the immune response to a standardized enterovirus challenge (polio) and infections not limited to the gastrointestinal tract in healthy adults . METHODS . In a randomized, controlled and double-blind study 64 volunteers consumed for 5 weeks chemically acidified clotted milk without bacteria or with 10(10)/serving (Lactobacillus rhamnosus) GG or Lactobacillus acidophilus CRL431 added . In the second week subjects were vaccinated orally against polio 1, 2 and 3 . Polio virus neutralizing serum activity, the primary parameter, was determined by the standard neutralization test (WHO) before and three times after vaccination . Polio-specific IgA, IgG and IgM were detected by ELISAs . RESULTS . Probiotics increased poliovirus neutralizing antibody titers (NT) and affected the formation of poliovirus-specific IgA and IgG in serum . The maximum increase after immunization was about 2, 2.2, or 4-fold higher, respectively, for NT, IgG or, IgA, in volunteers consuming probiotics instead of placebo . No consistent difference was noted between bacterial strains . CONCLUSIONS . Probiotics induce an immunologic response that may provide enhanced systemic protection of cells from virus infections by increasing production of virus neutralizing antibodies.

J Pharm Pharm Sci, 2004 Sep 24, 7(3), 315 - 24
Artificial cell microcapsule for oral delivery of live bacterial cells for therapy: design, preparation, and in-vitro characterization; Ouyang W et al.; PURPOSE: Bacterial cells can be engineered to synthesize a wide array of disease modifying substrates such as cytokines, vaccines and antibodies; however, their use as an orally delivered therapeutic is limited by poor gastrointestinal (GI) survival and instigation of immunogenic response . Artificial cell microcapsules have been well studied as a means to overcome such problems, however, presently obtainable microcapsules have limitations . This study summarizes a novel microcapsule design specifying its preparation and GI stability in-vitro . METHOD: Multilayer APPPA microcapsules were designed, prepared and characterized in-vitro for bacterial cell oral delivery using Lactobacillus reuteri cells as a model . Microcapsule structural integrity, mechanical stability, and GI survival studies were performed in simulated gastric (SGF) and intestinal (SIF) fluids in various pH conditions at 37.2 degrees C and compared with presently available alginate/poly-l-lysine/alginate (APA) microcapsules . HPLC was used for the microcapsule membrane permeability study . RESULTS: Results show that APPPA microcapsules can be prepared for bacterial cell encapsulation and are stable in simulated GI conditions . No microcapsule damage was reported when exposed to SGF and SIF for 12 hours at 250 rpm mechanical shaking at 37.2 degrees C . In addition, 93.2+/-2.3% and 98.9+/-0.6% of microcapsules were undamaged after 24 hours in SGF and SIF respectively . Microcapsule pH stability results show that 92.8+/-3.1% of microcapsules remained intact at pH 1, 3, 5, and 7 and no damage was observed at pH 9.0 when challenged for 24 hours . When exposed for 3 hours with 250 rpm shaking at 37.2 degrees C, no damage of the microcapsules in SGF and SIF at pH, 1,3,5,7, and 9 was observed . Compared to APA microcapsules, APPPA membranes showed superior GI stability and permeability for cell encapsulation . CONCLUSION: Novel APPPA microcapsules have superior features for oral delivery of live bacterial cells and they can be used for various clinical applications . However, further study such as membrane permeability, cytotoxicity, immune protection capacity, and suitability for live bacterial cell oral delivery in-vivo is required.

Appl Environ Microbiol, 2004 Dec, 70(12), 7260 - 8
Evidence for involvement of at least six proteins in adaptation of Lactobacillus sakei to cold temperatures and addition of NaCl; Marceau A et al.; Lactobacillus sakei is a lactic acid bacterium widely represented in the natural flora of fresh meat . The aim of this study was to analyze the differences in protein expression during environmental changes encountered during technological processes in which L . sakei is involved in order to gain insight into the ability of this species to grow and survive in such environments . Using two-dimensional electrophoresis, we observed significant variation of a set of 21 proteins in cells grown at 4 degrees C or in the presence of 4% NaCl . Six proteins could be identified by determination of their N-terminal sequences, and the corresponding gene clusters were studied . Two proteins belong to carbon metabolic pathways, and four can be clustered as general stress proteins . A phenotype was observed at low temperature for five of the six mutants constructed for these genes . The survival of four mutants during stationary phase at 4 degrees C was affected, and surprisingly, one mutant showed enhanced survival during stationary phase at low temperatures.

Appl Environ Microbiol, 2004 Dec, 70(12), 7200 - 9
Bile-mediated aminoglycoside sensitivity in Lactobacillus species likely results from increased membrane permeability attributable to cholic acid; Elkins CA et al.; Few studies have been conducted on antimicrobial resistance in lactobacilli, presumably because of their nonpathogenic nature as anaerobic commensals . We assessed resistance in 43 type strains and isolates representing 14 species by using agar disk diffusion and MIC analysis in MRS medium . Most noteworthy were two general phenotypes displayed by nearly every strain tested: (i) they were more susceptible (up to 256-fold in some cases) to the deconjugated bile acid cholic acid than to the conjugate taurocholic or taurodeoxycholic acid, and (ii) they became susceptible to aminoglycosides when assayed on agar medium containing 0.5% fractionated bovine bile (ox gall) . Two-dimensional MIC analyses of one representative strain, Lactobacillus plantarum WCFS1, at increasing concentrations of ox gall (0 to 30.3 mg/ml) displayed corresponding decreases in resistance to all of the aminoglycosides tested and ethidium bromide . This effect was clinically relevant, with the gentamicin MIC decreasing from >1,000 to 4 mug/ml in just 3.8 mg of ox gall per ml . In uptake studies at pH 6.5, {G-3H}gentamicin accumulation increased over control levels when cells of this strain were exposed to bile acids or reserpine but not when they were exposed to carbonyl cyanide m-chlorophenylhydrazone . The effect was dramatic, particularly with cholic acid, increasing up to 18-fold, whereas only modest increases, 3- and 5-fold, could be achieved with taurocholic acid and ox gall, respectively . Since L . plantarum, particularly strain WCFS1, is known to encode bile salt hydrolase (deconjugation) activity, our data indicate that mainly cholic acid, but not taurocholic acid, effectively permeabilizes the membrane to aminoglycosides . However, at pHs approaching neutral conditions in the intestinal lumen, aminoglycoside resistance due to membrane impermeability may be complemented by a potential efflux mechanism.

Appl Environ Microbiol, 2004 Dec, 70(12), 6992 - 7
Initiation of protein synthesis by a labeled derivative of the Lactobacillus casei DN-114 001 strain during transit from the stomach to the cecum in mice harboring human microbiota; Oozeer R et al.; Although studies on the survival of bacteria in the digestive tract have been reported in the literature, little data are available on the physiological adaptation of probiotics to the digestive environment . In previous work, a transcriptional fusion system (i.e., luciferase genes under the control of a deregulated promoter) was used to demonstrate that a derivative of the Lactobacillus casei DN-114 001 strain, ingested in a fermented milk and thus exhibiting initially a very weak metabolic activity, synthesized proteins de novo after its transit in the digestive tract of mice harboring human microbiota (known as human-microbiota-associated mice) . With the same genetic system and animal model, we here investigate for the first time the ability of L . casei to reinitiate synthesis in the different digestive tract compartments . In this study, most ingested L . casei cells transited from the stomach to the duodenum-jejunum within 1 h postingestion . No luciferase activity was observed in these digestive tract compartments after the first hour . At later times, the bulk of bacteria had transited to the ileum and the cecum . Luciferase synthesis was detected between 1.5 and 2.0 h postingestion at the ileal level and from 1.5 h to at least 6.0 h postingestion in the cecum, where the activity remained at a maximum level . These results demonstrate that ingested L . casei (derivative of the DN-114 001 strain) administered via a fermented milk has already reinitiated protein synthesis when it reaches the ileal and cecal compartments.

Med Sci Monit . 2004 Nov 24;10(12):CR662-666 {Epub ahead of print}
Effect of Lactobacillus casei supplementation on the effectiveness and tolerability of a new second-line 10-day quadruple therapy after failure of a first attempt to cure Helicobacter pylori infection; Tursi A et al.; Background: Probiotics have never been used as second-line treatment in patients resistant to a first course of anti-H . pylori treatment . Material/Methods: 70 consecutive patients with persistent H . pylori infection were enrolled and treated with ranitidine bismuth citrate (RBC) 400 mg b.d, esomeprazole or pantoprazole 40 mg/day, amoxycillin 1 g t.d, tinidazole 500 mg b.d . with (group A) or without (group B) supplementation with 750 mg daily containing 16 billion bacteria Lactobacillus casei subsp . casei DG . Esomeprazole or pantoprazole 40 mg/day was administered for a further 4 weeks in cases of active peptic ulcer or severe gastritis detected at endoscopy . In these cases endoscopy was repeated one month after conclusion of therapy . The remaining patients were checked by alC-urea breath test . Results: Sixty-six patients completed the study, 34 in group A and 32 in group B . One group A patient (2.85%) was excluded for protocol violation and one group B patient (2.85%) was lost to follow-up . 33/34 group A patients were H . pylori-negative {per-protocol: 97.05%, on intention-to-treat: 94.28%} . 5/34 patients (14.7%) showed side-effects, but all of them completed the treatment . In group B, two patients (5.71%) showed severe side-effects and were withdrawn from the study . 30/32 patients were H . pylori- negative {per-protocol: 93.75%, on intention-to-treat: 85.71% (p=n.s.)} . 11/32 patients (34.37%) showed side-effects, but all of them completed the study (p<0.05) . Conclusions: This 10-day quadruple therapy obtains a high eradication rate, but probiotic supplementation reduces side-effects and permits a slight improvement in eradicating H . pylori.

Expert Rev Anti Infect Ther, 2004 Dec, 2(6), 913 - 22
Evolving issues in understanding and treating bacterial vaginosis; Marrazzo JM; Bacterial vaginosis is a synergistic polymicrobial syndrome characterized by depletion of Lactobacillus spp., especially those that produce hydrogen peroxide, and an intense increase in the quantity of commensal vaginal anaerobic bacteria to 100- to 1000-fold above normal levels . While the bacterial spectrum of these organisms has long been known to include Gardnerella vaginalis, Prevotella spp., anaerobic Gram-positive cocci, Mobiluncus spp . and Mycoplasma hominis, innovative use of molecular diagnostics has identified novel species apparently associated with this syndrome, including Atopobium vaginalis . Effecting resolution of bacterial vaginosis is important, in particular for the 8 to 23% of women afflicted with symptomatic disease during their reproductive years . Bacterial vaginosis has been consistently associated with numerous adverse sequelae related to the upper genital tract, including pelvic inflammatory disease and postsurgical infection in the setting of invasive gynecologic procedures, and may increase women's risk of acquiring HIV infection . Pregnant women with bacterial vaginosis experience a higher rate of preterm delivery and low-birth-weight infants . While antibiotics with activity against anaerobes--typically, metronidazole and clindamycin applied vaginally or taken orally--are the mainstays of therapy, bacterial vaginosis frequently recurs . For these reasons, innovative approaches to therapy are urgently required.

J Chemother, 2004 Oct, 16(5), 463 - 7
Microbiological evaluation of commercial probiotic products available in Italy; Drago L et al.; Scientific evidence of the prevention and therapy of some intestinal diseases is accumulating in regard to probiotic products . However, sufficient information on the use of probiotics in specific therapies is not yet available and, above all, there is no clear legislation about these products in Europe . In this study, we evaluated five different probiotic products commercially available in Italy for their qualitative and quantitative microbial content after about 12 and 22 months of storage . We also evaluated the stability of lactobacilli to 0.3% bile salts and to pH of 3.58 and 7.98 . There were discrepancies between the declared content and our results found after storage for 4 of the tested products . Bile salts and basic pH did not affect the growth of the lactobacilli tested, while for 2 tested products 6 hours at acid pH produced a complete inhibition of bacterial growth . Our results suggest the need for clear legislation and adequate control of the manufacturing of probiotic products.

Eur J Dermatol, 2004 Nov-Dec, 14(6), 400 - 6
Clinical usefulness of oral itraconazole, an antimycotic drug, for refractory atopic dermatitis; Ikezawa Z et al.; We investigated the clinical usefulness of oral itraconazole for refractory atopic dermatitis in a crossover study . Patients with refractory atopic dermatitis were divided into two groups: Group A; a combination of itraconazole plus a conventional lactobacillus preparation was administered for 8 weeks, followed by lactobacillus preparation alone for 8 weeks, Group B; lactobacillus preparation alone was administered for 8 weeks, followed by itraconazole plus lactobacillus for 8 weeks . In both groups, a decrease in dose or strength of concomitant topical steroids was observed at the end of the treatment course of itraconazole, and improvement of parameters such as eosinophil count, serum IgE level and specific IgE antibody titers to fungi was also observed after the administration of itraconazole . These results suggest that oral itraconazole is useful for the treatment of intractable atopic dermatitis patients who do not respond to conventional therapeutic approaches.

J Agric Food Chem, 2004 Dec 1, 52(24), 7300 - 5
Survival of freeze-dried Lactobacillus bulgaricus KFRI 673 in chitosan-coated calcium alginate microparticles; Lee JS et al.; The aim of this study was to investigate the effect of alginate microparticles coated with three kinds of chitosans of different molecular weights on the survival of Lactobacillus bulgaricus KFRI 673 in simulated gastric (SGJ) and intestinal juices (SIJ) and on their stability during storage at 4 and 22 degrees C . L . bulgaricus KFRI 673 loaded in alginate microparticles was prepared by spraying the mixture of sodium alginate and cell culture into the calcium chloride solution using an air-atomizing device . When L . bulgaricus KFRI 673 was exposed to SGJ of pH 2.0 for 60 min, none of the microorganism survived . Contrary to this result, microbiological analysis indicated that microencapsulation in alginate microparticles improved the survival of acid-sensitive L . bulgaricus KFRI 673 in SGJ and that high molecular weight chitosan coating resulted in the highest survival in SGJ . To study storage stability of free and microencapsulated cells, in vitro studies were conducted at 4 and 22 degrees C during a 4 week period . Both free and microencapsulated cells showed similar stabilities during 4 weeks of storage at 4 degrees C . However, the stability of Lactobacillus at 22 degrees C was appreciably improved when loaded in high molecular weight chitosan-coated alginate microparticles . In conclusion, microencapsulation of lactic acid bacteria with alginate and chitosan coating offers an effective way of delivering viable bacterial cells to the colon and maintaining their survival during refrigerated storage.

East Mediterr Health J, 2003 Jan-Mar, 9(1-2), 141 - 7
Effect of natural starters used for sourdough bread in Morocco on phytate biodegradation; Chaoui A et al.; Phytase activity was studied in natural sourdough bread starters to determine physicochemical characteristics (phytic acid hydrolysis, dough rising capacity and pH) in the flour and during sourdough fermentation . Fermentation microorganisms (yeasts and lactic acid bacteria) were also characterized . Results showed a decrease of phytic acid in sourdoughs started with traditional starters, and wide variation in phytase activity . Microorganism counts were high at the end of fermentation, indicating higher fermenting activity of the starters . Yeast populations showed wide variation and lactic acid bacteria had high counts in the fermentation . Phytase activity was demonstrated in starter cultures made of lactic acid bacteria and yeast isolates, the most interesting of which were Saccharomyces cerevisiae combined with Lactobacillus plantarum and Leuconostoc mesenteroides.

Clin Invest Med, 2004 Oct, 27(5), 274 - 81
Probiotics in surgical wound infections: current status; Howard JC et al.; BACKGROUND: Probiotics--live microorganisms that confer a health benefit when taken in adequate amounts, usually as food supplements--are receiving renewed attention in the medical community . Some have been found to play a role in disease remediation . However, mainstream medicine and science remain divided about the validity of health claims made about them . METHODS: To clarify the potential value of probiotics, we reviewed the scientific data on their role in preventing and treating surgical infections as well as some of our own studies of the effects of certain strains of lactobacilli on surgical implant infections . PRINCIPAL FINDINGS: There is little rigorous evidence that probiotics may be beneficial in the prevention and treatment of wound infections . However, data from 3 clinical trials and from our laboratory indicate that certain strains of probiotic lactobacilli and their byproducts may help reduce infection rates in surgical patients and may ameliorate staphylococcus-related infections of surgical implants . CONCLUSION: Although there is good clinical evidence that certain probiotics may be beneficial in conditions such as diarrheal and inflammatory bowel diseases, more studies are required to apply these concepts to the prevention and treatment of wound and other surgical infections.

Rev Argent Microbiol, 2004 Jul-Sep, 36(3), 139 - 44
{Microbiological study of garlic (Allium sativum L.) and onion (Allium cepa L.) dehydrated}; Fuselli SR et al.; A microbiological study during the process and the storage of garlic (Allium sativum L.) and onion (Allium cepa L.) dehydrated, with the additional barriers of blanching or brine immersion, was made . In all raw materials the average counts of aerobic mesophilic bacteria expressed in CFU/g ranged from 1.2 x 10(2) to 1.6 x 10(3), molds and yeasts from 60 to 1.6 x 10(3), Lactobacillus spp . and Leuconostoc mesenteroides subsp . mesenteroides between 10 and 50 . Microorganisms identified were Penicillium spp., Monilia spp., Lactobacillus brevis, Leuconostoc mesenteroides subsp . mesenteroides and yeasts in garlic; Mucor spp., Penicillium spp., Monilia spp., Lactobacillus brevis and yeasts in both types of onions . Leuconostoc mesenteroides subsp . mesenteroides was detected in only kind of onion . In dehydrated garlic storage, Penicillium spp., Monilia spp., Lactobacillus brevis and yeasts were detected . In garlic, when a blanching step was carried out no microflora was detected . Mucor spp., Penicillium spp., Monilia spp . and Lactobacillus brevis were identified in both types of dehydrated onions . When brine immersion was included the microflora detected was significantly lower and only Penicillium spp . were found . The use of additional barriers such as blanching or brine immersion produces an important effect on the microbiological stability in these products.

Appl Microbiol Biotechnol, 2004 Dec, 66(2), 160 - 5 Epub 2004 Dec.
Production of L-lactic acid from a mixture of xylose and glucose by co-cultivation of lactic acid bacteria; Taniguchi M et al.; The production of optically pure lactic acid in a high yield from xylose or a mixture of xylose and glucose, which is a model hydrolysate of lignocellulose, is described . In a single cultivation, Enterococcus casseliflavus produced 38 g/l of lactic acid with an optical purity of 96% enantiomeric excess (ee) and 6.4 g/l of acetic acid from 50 g/l of xylose when MRS medium was used . When a mixture of 50 g/l of xylose and 100 g/l of glucose was used as the carbon source in a cultivation of E . casseliflavus alone, glucose was converted to lactic acid in the early phase of the cultivation but xylose was hardly consumed . In a co-cultivation where E . casseliflavus and Lactobacillus casei specific for glucose were simultaneously inoculated, little or no lactic acid was produced after the glucose was almost consumed . A co-cultivation with two-stage inoculation (in which E . casseliflavus was added at a cultivation time of 40 h after L . casei cells were inoculated) resulted in complete consumption of 50 g/l of xylose and 100 g/l of glucose . In the co-cultivation, 95 g/l of lactic acid with a high optical purity of 96% ee was obtained at 192 h . Such a co-cultivation using two microorganisms specific for each sugar is considered to be one promising cultivation technique for the efficient production of lactic acid from a sugar mixture derived from lignocellulose.

FEMS Microbiol Lett, 2004 Dec 1, 241(1), 73 - 7
Transformation of Lactobacillus plantarum by electroporation with in vitro modified plasmid DNA; Alegre MT et al.; An improved method for the electrotransformation of Lactobacillus plantarum CECT 220 (ATCC 8014) with plasmid DNA isolated from Escherichia coli is described . The two main modifications with respect to existing methods are: (i) isolation of plasmid DNA from E . coli JM110 grown in minimal medium and (ii) in vitro modification of the DNA by cell-free extracts of the host L . plantarum . Optimal electrotransformation was obtained with exponentially growing cells of L . plantarum concentrated to 6x10(9) cfu ml-1, with electric pulses of 13 kV cm-1 in cuvettes with 1 mm inter-electrode distance . We consider that this method constitutes a useful tool for routine manipulation of L . plantarum, and can probably be extended to other lactic acid bacteria.

Int J Antimicrob Agents, 2004 Dec, 24(6), 578 - 84
Porphyrin-based compounds exert antibacterial action against the sexually transmitted pathogens Neisseria gonorrhoeae and Haemophilus ducreyi; Bozja J et al.; A series of porphyrin based compounds without (nMP) or with (MP) metals were found to have potent bactericidal action in vitro against the sexually transmitted pathogens Neisseria gonorrhoeae and Haemophilus ducreyi . nMP and MP did not show bactericidal activity against five species of lactobacilli . An MP containing gallium had the capacity to block a gonococcal infection in a murine vaginal model, indicating that its development as a topical microbicide to block sexually transmitted bacterial infections is warranted . In contrast to other bacterial species, loss of the gonococcal haemoglobin uptake system encoded by hpuB or energy supplied through the TonB-ExbB-ExbD system did not significantly affect levels of MP-susceptibility in gonococci . In contrast, mutations in gonococci that inactivate the mtrCDE-encoded efflux pump were found to enhance gonococcal susceptibility to nMPs and MPs while over-production of this efflux pump decreased levels of gonococcal susceptibility to these compounds.

Zh Mikrobiol Epidemiol Immunobiol, 2004 Sep-Oct, (5), 94 - 8
{Quantitative characterization of antagonistic activity of lactobacilli}
{Comparative study of monosaccharide composition of group Bacillus--Lactobacillus--Streptococcus bacteria glycocalix by biochemical and electron-microscopic methods}
{No authors listed}

Results of comparison of the data on monosaccharide composition of glycocalix in group Bacillus--Lactobacillus--Streptococcus bacteria using biochemical and electronmicroscopic methods prove that the composition and quantity of monosaccharides varied depending on the genus and peculiarities of microorganisms strains (biological activity in respect of people and warm-blooded animals) . At the same time, representatives of this group of microorganisms in the glycocalix had common monosaccharides: glucosamine, sialic acids, galactosamine, galactose, glucose, mannose, fucose, that is the phenotypical display of genetic relations.

Int J Syst Bacteriol, 1996 Apr, 46(2), 595 - 8
Lactobacillus paraplantarum sp . now., a new species related to Lactobacillus plantarum; Curk MC et al.; Four strains of facultatively heterofermentative lactobacilli isolated from beer and human feces have physiological characteristics similar to those of Lactobacillus plantarum . Unlike 66% of the L . plantarum strains tested (F . Bringel, M.-C . Curk, and J.-C . Hubert, Int . J . Syst . Bacteriol . 46:588-594, 1996), these strains do not catabolize alpha-methyl-D-mannoside . However, because they exhibit little DNA relatedness to L . plantarum and Lactobacillus pentosus, these four strains were classified as members of a new species, Lactobacillus paraplantarum; strain CNRZ 1885 (= CIP 104668) is the type strain.

Appl Microbiol Biotechnol . 2004 Nov 12; {Epub ahead of print}
Enantioselective reduction of carbonyl compounds by whole-cell biotransformation, combining a formate dehydrogenase and a ( R)-specific alcohol dehydrogenase; Ernst M et al.; A whole-cell biotransformation system for the reduction of prochiral carbonyl compounds, such as methyl acetoacetate, to chiral hydroxy acid derivatives {methyl ( R)-3-hydroxy butanoate} was developed in Escherichia coli by construction of a recombinant oxidation/reduction cycle . Alcohol dehydrogenase from Lactobacillus brevis catalyzes a highly regioselective and enantioselective reduction of several ketones or keto acid derivatives to chiral alcohols or hydroxy acid esters . The adh gene encoding for the alcohol dehydrogenase of L . brevis was expressed in E . coli . As expected, whole cells of the recombinant strain produced only low quantities of methyl ( R)-3-hydroxy butanoate from the substrate methyl acetoacetate . Therefore, the fdh gene from Mycobacterium vaccae N10, encoding NAD(+)-dependent formate dehydrogenase, was functionally coexpressed . The resulting two-fold recombinant strain exhibited an in vitro catalytic alcohol dehydrogenase activity of 6.5 units mg(-1) protein in reducing methyl acetoacetate to methyl ( R)-3-hydroxy butanoate with NADPH as the cofactor and 0.7 units mg(-1) protein with NADH . The in vitro formate dehydrogenase activity was 1.3 units mg(-1) protein . Whole resting cells of this strain catalyzed the formation of 40 mM methyl ( R)-3-hydroxy butanoate from methyl acetoacetate . The product yield was 100 mol% at a productivity of 200 mumol g(-1) (cell dry weight) min(-1) . In the presence of formate, the intracellular {NADH}/{NAD(+)} ratio of the cells increased seven-fold . Thus, the functional overexpression of alcohol dehydrogenase in the presence of formate dehydrogenase was sufficient to enable and sustain the desired reduction reaction via the relatively low specific activity of alcohol dehydrogenase with NADH, instead of NADPH, as a cofactor.

J Bacteriol, 2004 Dec, 186(23), 7829 - 35
Genetic characterization of the bile salt response in Lactobacillus plantarum and analysis of responsive promoters in vitro and in situ in the gastrointestinal tract; Bron PA et al.; In this paper we describe the growth, morphological, and genetic responses of Lactobacillus plantarum WCFS1 to bile . Growth experiments revealed that a stepwise increase in the porcine bile concentration led to a gradual decrease in the maximal growth rate . Moreover, the final density reached by an L . plantarum culture growing in MRS containing 0.1% bile was approximately threefold lower than that in MRS lacking bile . The morphology of the cells grown in MRS containing 0.1% bile was investigated by scanning electron microscopy, which revealed that cells clumped together and had rough surfaces and that some of the cells had a shrunken and empty appearance, which clearly contrasted with the characteristic rod-shaped, smooth-surface morphology of L . plantarum cells grown in MRS without bile . An alr complementation-based genome-wide promoter screening analysis was performed with L . plantarum, which led to identification of 31 genes whose expression was potentially induced by 0.1% porcine bile . Remarkably, 11 membrane- and cell wall-associated functions appeared to be induced by bile, as were five functions involved in redox reactions and five regulatory factors . Moreover, the lp_0237 and lp_0775 genes, identified here as genes that are inducible by bile in vitro, were previously identified in our laboratory as important for L . plantarum in vivo during passage in the mouse gastrointestinal tract (P . A . Bron, C . Grangette, A . Mercenier, W . M . de Vos, and M . Kleerebezem, J . Bacteriol . 186:5721-5729, 2004) . A quantitative reverse transcription-PCR approach focusing on these two genes confirmed that the expression level of lp_0237 and lp_0775 was significantly higher in cells grown in the presence of bile and cells isolated from the mouse duodenum than in cells grown on laboratory medium without bile.

J Appl Microbiol, 2004, 97(6), 1297 - 310
Isolation of a lactic acid bacterium and yeast consortium from a fermented material of Ulva spp . (Chlorophyta); Uchida M et al.; AIMS: Microbiota in a fermented culture of Ulva spp . was examined with the objective to characterize the type of fermentation and to obtain starter microbes for performing seaweed fermentation . METHOD AND RESULTS: Fermented Ulva spp . cultures which were obtained and transferred in a laboratory were examined for their microbiota . With phenotypic characterization and phylogenetic analysis based on rRNA gene nucleotide sequences, the predominant micro-organisms were identified as Lactobacillus brevis, Debaryomyces hanseni var . hansenii, and a Candida zeylanoides-related specimen, suggesting that the observed fermentation can be categorized to lactic acid and ethanol fermentation . Inoculating the individually cultured cell suspensions of the three kinds of micro-organisms with cellulase induced the fermentation in various kinds of seaweed . CONCLUSIONS: A microbial consortium composed of a lactic acid bacterium, L . brevis, and yeasts, D . hansenii and a C . zeylanoides-related specimen, were predominant in a fermented culture of Ulva spp . Lactic acid and ethanol fermentation could be induced in various kinds of seaweed by adding this microbial consortium along with cellulase . SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of lactic acid and ethanol fermentation in seaweed, which is expected to provide a new material for food and dietary applications.

J Dairy Sci, 2004 Dec, 87(12), 4097 - 103
Purification and characterization of intracellular proteinase from Lactobacillus casei ssp . casei LLG; Shin JY et al.; The intracellular proteinase of Lactobacillus casei ssp . casei LLG was isolated in the cytoplasmic fraction with 0.05 M Tris-HCl buffer (pH 7.5) . The enzyme was purified by the fast protein liquid chromatography system equipped with ion-exchange and gel filtration chromatographies . This proteinase comprised a single monomeric form and had a molecular weight of about 55 kDa and an isoelectric point near pH 4.9 . The optimum pH and temperature for the enzyme activity were determined to be pH 6.5 and 37 degrees C, respectively . The enzyme was inactivated by metal-chelating compounds (EDTA, 1,10-phenanthroline) and less affected by serine proteinase inhibitors (diisopropylfluorophosphate, phenylmethylsulfonyl fluoride) . Proteinase activity was increased by Ca++, Mn++, and Co++, and inhibited by Cu++, Mg++, and Zn++ . The activity of this enzyme to hydrolyze casein appeared to be more active on beta-casein than alphas1-casein and kappa-casein as monitored by polyacrylamide gel electrophoresis.

Int J Pharm, 2004 Dec 9, 287(1-2), 163 - 73
Nisin-loaded poly-L-lactide nano-particles produced by CO2 anti-solvent precipitation for sustained antimicrobial activity; Salmaso S et al.; Nisin-loaded poly-L-lactide (PLA) nano-particles were fabricated by processing protein/polymer organic solutions by semi-continuous compressed CO2 anti-solvent precipitation . Preliminary solubility studies were carried out for an optimised selection of organic solvent mixtures leading to preparation of protein/polymer solutions . The particles were prepared by processing 50:50 dimethylsulfoxide/dichloromethane mixtures containing 1% polymer (w/v) and 5 or 20% nisin (nisin/polymer, w/w) . Proper operative conditions (organic solution injection rate, precipitation temperature and gas pressure, CO2 flow rate, washing time, etc.) were set up to yield production reproducibility, high product recovery (over 70%) and high drug loading (over 95% of the recovered protein) . Scanning electron microscopy demonstrated that spherical, smooth surfaced particles were produced . Light scattering showed that the particle size was in the range of 200-400 nm and the products were characterised by narrow polydispersity . In vitro release studies showed that the protein is slowly released throughout 1000 h . However, the release was slower as the salt concentration and the pH of the release buffer increased . Solubility investigations suggested that the observed differences in protein release rate out of nano-particles was attributable to the protein interaction with the polymer which was found to increase as the pH or the salt concentration increased in the release buffer . In vitro studies carried out by nano-particle incubation in medium containing Lactobacillus delbrueckeii showed that nisin was released in the active form and the antibacterial activity was maintained up to 45 days incubation.

Int J Food Microbiol, 2004 Dec 15, 97(2), 147 - 56
Numbers and strains of lactobacilli in some probiotic products; Coeuret V et al.; The numbers and types of Lactobacillus strains in probiotic feed or food products for sale in Europe were compared with the information stated on the product labels . The labels of all 10 products gave information on the genus and species of Lactobacillus, and seven gave information on the numbers of a specified strain(s) . Lactobacilli were recovered using a recently validated method, developed as part of the European Community Project SMT4 CT98-2235 for the official control of probiotic microorganisms used as feed additives . Polymerase chain reaction (PCR)-based methods were used to assess the accuracy of labelling with regard to genus and species, and pulsed-field gel electrophoresis (PFGE) was used to identify strains . Five products were mislabelled with respect to the numbers and three with respect to species of lactobacilli . In four cases, the specified strains were not detected . Four fermented milks sold under three trademarks contained the same strain, which was named differently on each label . As safety and functionality of probiotics are strain dependent, these results demonstrated the need to control lactobacilli present in commercially probiotic human food products, not only at the species but also at the strain level, to ensure their quality and protect the consumer . The usefulness of the official methods developed for animal feed additives to identify lactobacilli in probiotic foods for humans was demonstrated.

Vet Immunol Immunopathol, 2004 Dec 28, 102(4), 379 - 88
Immune responses in rainbow trout Oncorhynchus mykiss induced by a potential probiotic bacteria Lactobacillus rhamnosus JCM 1136; Panigrahi A et al.; This study was undertaken to examine the effect of supplementing a suggested probiotic bacteria Lactobacillus rhamnosus JCM 1136 in feed on immune response and gut flora composition of rainbow trout Oncorhynchus mykiss . The probiotic bacteria were incorporated into a commercial feed to constitute two experimental diets containing either 10(9) or 10(11) colony forming unit of live bacteria/g of feed while a third diet without the bacterial supplement served as the control diet . The diets were offered to rainbow trout (75g average weight) in triplicate tanks for 30 days . Fish were sampled at 10, 20 and 30 days after commencement of the feeding trial to determine the proportion of the given probiont in the gut microflora composition and the nonspecific humoral and cellular immune responses on the 30th day . The relative proportion of the probiont increased with the feeding duration in the intestine, but not in the stomach . The proportion of L . rhamnosus in the stomach corresponded to the intake levels while no such relation existed in the intestine . The serum lysozyme and complement activities were significantly greater in fish fed the higher level of probiont compared with the control fish . The phagocytic activity of head kidney leucocytes also showed similar tendencies . These observations indicate the potential immuno-regulatory role of probiotic organisms in rainbow trout.

Clin Diagn Lab Immunol, 2004 Nov, 11(6), 1171 - 81
Induction of a humoral immune response following an Escherichia coli O157:H7 infection with an immunomodulatory peptidic fraction derived from Lactobacillus helveticus-fermented milk; Leblanc J et al.; Numerous beneficial effects have been attributed to probiotic lactic acid bacteria (LAB), such as the stimulation of the immune system, the prevention of enteric infections by enteropathogens, and the regression of immunodependent tumors . It has been shown that biologically active metabolites released during fermentation, in particular biopeptides, could act as immunomodulatory agents . However, no studies have been conducted to evaluate the implication of these bioactive peptides in the induction of a protective immune response against enteric infections . The present study aimed to evaluate the possible immunomodulatory and anti-infectious effects of a peptidic fraction released in milk fermented by Lactobacillus helveticus . The immune response in the mucosa-associated lymphoid tissue was monitored following an administration of the potentially bioactive peptidic fraction . The total immunoglobulin A (IgA) immune response was evaluated after an Escherichia coli O157:H7 infection in a BALB/c murine model . Immunohistochemical and enzyme-linked immunosorbent assays revealed an increase in the number of IgA-secreting B lymphocytes in the intestinal lamina propria and an enhanced total secretory and systemic IgA response . Cytokine profiling also revealed stimulation of a Th2 response in mice fed the peptidic fraction, whereas infected controls demonstrated a proinflammatory Th1 response . These results indicate that bioactive peptides released during fermentation by LAB could contribute to the known immunomodulatory effects of probiotic bacteria.

J Agric Food Chem, 2004 Nov 17, 52(23), 6923 - 31
Identification of angiotensin-I-converting enzyme inhibitory peptides derived from sodium caseinate hydrolysates produced by Lactobacillus helveticus NCC 2765; Robert MC et al.; Angiotensin-I-converting enzyme (ACE) inhibitory activity was identified in milk proteins fermented with Lactobacillus (Lb.) helveticus NCC 2765 (Nestle Culture Collection, Vers-chez-les-Blanc, Switzerland) . Hydrolyzing sodium caseinate for 1 and 2 h inhibited ACE activity, as measured by an in vitro ACE inhibition test . The hydrolysates with the highest ACE inhibitory potential were fractionated by gel permeation chromatography and their low molecular weight fractions collected . These fractions were subsequently subfractionated by reverse-phase high-pressure liquid chromatography . Several hydrophobic subfractions showed high ACE inhibitory potential, and their peptide composition was determined using an ion trap mass spectrometer equipped with an elctrospray ionization source . Analysis of the low molecular weight fraction identified 14 peptides with known antihypertensive activity and 1 with previously described opioid activity . On the basis of the peptide composition of active subfractions, two potentially active novel sequences were defined, and the following synthetic peptides were synthesized: FVAPFPEVFG (alphaS1 39-48), ENLLRFFVAPFPEVFG (alphaS1 33-48), NENLLRFFVAPFPEVFG (alphaS1 32-48), LNENLLRFFVAPFPEVFG (alphaS1 31-48), NLHLPLPLL (beta 147-155), ENLHLPLPLL (beta 146-155), and VENLHLPLPLL (beta 145-155) . The ACE inhibitory potential of these synthetic peptides was assessed, and IC50 values were determined . NLHLPLPLL (beta 147-155), which was the only synthetic peptide also present in the sodium caseinate hydrolysates, and NENLLRFFVAPFPEVFG (alphaS1 32-48) showed the highest inhibition of ACE activity, with IC50 values of 15 and 55 microM, respectively . Furthermore, the stability of all synthetic peptides was assessed using an in vitro model simulating gastric digestion . The beta-casein-derived peptides remained intact following the successive hydrolysis by pepsin and pancreatin, whereas alphaS1-casein-derived peptides were degraded by pepsin.

SADJ . 2004 Aug;59(7):280, 282.
Anti-bacterial efficacy of new self-etching dentine bonding agent; Brandt PD et al.; Since a perfect seal is not easily achieved during bonding procedures, any product which will limit or delay bacterial proliferation in the resulting micro-leakage spaces should extend the life span of bonded restorations . This study compared the antibacterial properties of an experimental, self-etching, fluoride-releasing, antibacterial bonding system (ABF) with two standard bonding systems (SE Bond and Scotchbond MP) . Spread plates of three different bacteria (Streptococcus mutans, Lactobacillus paracasei and Actinomyces naeslundii) were prepared on Brain-Heart Infusion agar . Standardised, sterilised filtration paper disks were infiltrated with 20 microl of primer and then placed on the inoculated agar and incubated at 37 degrees C for 48 hours . The extent of the inhibition zones were measured at different positions and data were analysed using the Student t-test to determine significant differences . All three primers showed zones of inhibition for all three bacteria tested . Inhibition zones for ABF primer against S . mutans were significantly larger (p<0.05) compared to that of Scotchbond MP and SE Bond . In general the antibacterial activity of the three primers against the three bacteria tested varied, and the inhibitory effect for the experimental primer was significantly superior against S . mutans only.

Med Hypotheses, 2005, 64(1), 64 - 8
Effect of antibiotics, prebiotics and probiotics in treatment for hepatic encephalopathy; Bongaerts G et al.; In order to reduce ammonia production by urease-positive bacteria Solga recently hypothesised (S.F . Solga, Probiotics can treat hepatic encephalopathy, Medical Hypotheses 2003; 61: 307-13), that probiotics are new therapeutics for hepatic encephalopathy (HE), and that they may replace antibiotics and lactulose . This influenced our view of the effect of antibiotics, prebiotics, e.g., lactulose, and probiotics on intestinal bacteria in the treatment of HE . Intestinal ammonia arises from aminoacids after bacterial de-amination and not from urea making urease-positive bacteria irrelevant . Antibiotics are not preferred in the treatment of HE, since ammonia-producing antibiotic-resistant bacteria may survive and replace ammonia-producing antibiotic-susceptible bacteria . Intestinal prebiotics are carbohydrate-like compounds, such as lactulose and resistant starch, that beneficially affects host's health in a different manner than normal food . In the small bowel prebiotics are not absorbed and digested, but are fermented in the colon by colonic bacteria . Fermentation of prebiotics yields lactic, acetic and butyric acids, as well as gas especially hydrogen (H2) . The massive H2 volumes cause rapid intestinal hurry and thus massive amounts of colonic bacteria, not only urease-positive bacteria, but also deaminating bacteria, are removed and intestinal uptake of toxic bacterial metabolites, e.g., ammonia, reduced . As living non-pathogenic micro-organisms, probiotics beneficially affect the host's health by fermenting non-absorbed sugars, especially in the small bowel . Thus, they reduce the substrate of the other bacteria, and simultaneously they create a surplus of fermentation products which may affect the non-probiotic flora . Regarding the fermentation products (lactic acid, ethanol, acetic acid and CO2) five groups of probiotic micro-organisms are known . It is argued that probiotic, CO2-producing (facultatively) heterolactic lactobacilli, i.e., lactobacilli, that produce both lactic acid and CO2 from sugars, such as glucose, are preferred in the treatment of HE . Our ideas concur with the practice guidelines regarding HE as formulated by Blei, Cordoba and the Practice Parameters Committee of the American College of Gastroenterology, and does not alter the final conclusion of Solga as regards the beneficial use in future treatment of HE.

Biotechnol Bioeng, 2004 Dec 20, 88(6), 681 - 9
Strain improvement and metabolic flux analysis in the wild-type and a mutant Lactobacillus lactis strain for L(+)-lactic acid production; Bai DM et al.; The effects of initial glucose concentration and calcium lactate concentration on the lactic acid production by the parent strain, Lactobacillus lactis BME5-18, were studied . The results of the experiments indicated that glucose and lactate repressed the cell growth and the lactic acid production by Lactobacillus lactis BME5-18 . A L(+)-lactic acid overproducing strain, Lactobacillus lactis BME5-18M, was screened by mutagenizing the parent strain with ultraviolet (UV) light irradiation and selecting the high glucose and lactate calcium concentration repression resistant mutant . Starting with a concentration of 100g L(-1) glucose, the mutant produced 98.6 g L(-1) lactic acid after 60 h in flasks, 73.9% higher than that of the parent strain . The L(+)-lactic acid purity was 98.1% by weight based on the amount of total lactic acid . The culture of the parent strain could not be analyzed well by conventional metabolic flux analysis techniques, since some pyruvate were accumulated intracellularly . Therefore, a revised flux analysis method was proposed by introducing intracellular pyruvate pool . Further studies demonstrate that there is a high level of NADH oxidase activity (12.11 mmol mg(-1) min(-1)) in the parent strain . The molecular mechanisms of the strain improvement were proposed, i.e., the high level of NADH oxidase activity was eliminated and the uptake rate of glucose was increased from 82.1 C-mmol (g DW h)(-1) to 98.9 C-mmol (g DW h)(-1) by mutagenizing the parent strain with UV, and therefore the mutant strain converts mostly pyruvate to lactic acid with a higher productivity (1.76 g L(-1) h(-1)) than the parent strain (0.95 g L(-1) h(-1)).

Clin Pharmacokinet, 2004, 43(14), 951 - 62
Enteral drug absorption in patients with short small bowel : a review; Severijnen R et al.; Drug therapy may become difficult when a significant amount of the small intestine is resected, as happens in patients with a short small bowel . Drug absorption from the gastrointestinal tract is altered in these patients; however, this effect is variable in patients and differs with each drug . Literature regarding clinical outcomes of normal or alternative administration routes in patients with a short small bowel is limited . We explored what is written about the normal absorption of commonly used drugs and what difference the resection of different but substantial parts of the small intestine makes . Changes in the gastrointestinal tract after resection of >50% of the small intestine causes malabsorption of macronutrients and micronutrients, and may alter the drug absorption process . The metabolic activity of the abundantly present intestinal lactobacilli can also affect the enteral drug absorption in patients with short small bowel as this results in the production of lactic acid, gaseous CO(2), ethanol and an increased bile acid deconjugation . Accelerated intestinal luminal transit time causes a reduction in absorption of certain antimicrobial agents, digoxin, hydrochlorothiazide, ciclosporin, cimetidine, mesalazine (5-aminosalicylic acid), oral contraceptives and levothyroxine . Gastric hypersecretion and lack of sufficient contact time with the intestinal mucosa in patients with short small bowel leads to insufficient absorption of drugs such as omeprazole . Successful treatment with warfarin, tricyclic antidepressants, metronidazole, fluconazole, procainamide, sotalol and pindolol are reported in several studies . Many different factors cause this variability in drug absorption in such patients . Monitoring the serum drug concentration in these patients may ease dealing with the management problems.

Macromol Biosci, 2004 Nov 20, 4(11), 1021 - 7
Production of D-lactic acid by bacterial fermentation of rice starch; Fukushima K et al.; D-Lactic acid was synthesized by the fermentation of rice starch using microorganisms . Two species: Lactobacillus delbrueckii and Sporolactobacillus inulinus were found to be active in producing D-lactic acid of high optical purity after an intensive screening test for D-lactic acid bacteria using glucose as substrate . Rice powder used as the starch source was hydrolyzed with a combination of enzymes: alpha-amylase, beta-amylase, and pullulanase to obtain rice saccharificate consisting of maltose as the main component . Its average gross yield was 82.5% . Of the discovered D-lactic acid bacteria, only Lactobacillus delbrueckii could ferment both maltose and the rice saccharificate . After optimizing the fermentation of the rice saccharificate using this bacterium, pilot scale fermentation was conducted to convert the rice saccharificate into D-lactic acid with a D-content higher than 97.5% in a yield of 70% . With this yield, the total yield of D-lactic acid from brown rice was estimated to be 47%, which is almost equal to the L-lactic acid yield from corn . The efficient synthesis of D-lactic acid can open a way to the large scale application of high-melting poly(lactic acid) that is a stereocomplex of poly(L-lactide) and poly(D-lactide) . Schematic representation of the production of D-lactic acid starting from brown rice as described here.

Microbiology, 2004 Nov, 150(Pt 11), 3681 - 90
Glucan synthesis in the genus Lactobacillus: isolation and characterization of glucansucrase genes, enzymes and glucan products from six different strains; Kralj S et al.; Members of the genera Streptococcus and Leuconostoc synthesize various alpha-glucans (dextran, alternan and mutan) . In Lactobacillus, until now, the only glucosyltransferase (GTF) enzyme that has been characterized is gtfA of Lactobacillus reuteri 121, the first GTF enzyme synthesizing a glucan (reuteran) that contains mainly alpha-(1-->4) linkages together with alpha-(1-->6) and alpha-(1-->4,6) linkages . Recently, partial sequences of glucansucrase genes were detected in other members of the genus Lactobacillus . This paper reports, for the first time, isolation and characterization of dextransucrase and mutansucrase genes and enzymes from various Lactobacillus species and the characterization of the glucan products synthesized, which mainly have alpha-(1-->6)- and alpha-(1-->3)-glucosidic linkages . The four GTF enzymes characterized from three different Lb . reuteri strains are highly similar at the amino acid level, and consequently their protein structures are very alike . Interestingly, these four Lb . reuteri GTFs have relatively large N-terminal variable regions, containing RDV repeats, and relatively short putative glucan-binding domains with conserved and less-conserved YG-repeating units . The three other GTF enzymes, isolated from Lactobacillus sakei, Lactobacillus fermentum and Lactobacillus parabuchneri, contain smaller variable regions and larger putative glucan-binding domains compared to the Lb . reuteri GTF enzymes.

Microbiology, 2004 Nov, 150(Pt 11), 3601 - 11
The complete genomes of Lactobacillus plantarum and Lactobacillus johnsonii reveal extensive differences in chromosome organization and gene content; Boekhorst J et al.; The first comprehensive comparative analysis of lactobacilli was done by comparing the genomes of Lactobacillus plantarum (3.3 Mb) and Lactobacillus johnsonii (2.0 Mb) . L . johnsonii is predominantly found in the gastrointestinal tract, while L . plantarum is also found on plants and plant-derived material, and is used in a variety of industrial fermentations . The L . plantarum and L . johnsonii chromosomes have only 28 regions with conservation of gene order, totalling about 0.75 Mb; these regions are not co-linear, indicating major chromosomal rearrangements . Metabolic reconstruction indicates many differences between L . johnsonii and L . plantarum: numerous enzymes involved in sugar metabolism and in biosynthesis of amino acids, nucleotides, fatty acids and cofactors are lacking in L . johnsonii . Major differences were seen in the number and types of putative extracellular proteins, which are of interest because of their possible role in host-microbe interactions . The differences between L . plantarum and L . johnsonii, both in genome organization and gene content, are exceptionally large for two bacteria of the same genus, emphasizing the difficulty in taxonomic classification of lactobacilli.

Appl Environ Microbiol, 2004 Nov, 70(11), 6936 - 9
Selection of single-chain antibodies against the VP8* subunit of rotavirus VP4 outer capsid protein and their expression in Lactobacillus casei; Monedero V et al.; Single-chain antibodies (scFv) recognizing the VP8* fraction of rotavirus outer capsid and blocking rotavirus infection in vitro were isolated by phage display . Vectors for the extracellular expression in Lactobacillus casei of one of the scFv were constructed . L . casei was able to secrete active scFv to the growth medium, showing the potential of probiotic bacteria to be engineered to express molecules suitable for in vivo antirotavirus therapies.

Gene, 2004 Nov 10, 342(1), 189 - 97
Identification and phenotypic characterization of the cell-division protein CdpA; Altermann E et al.; Analysis of the automated computer annotation of the early draft phase genome of Lactobacillus acidophilus NCFM revealed the previously discovered S-layer gene slpA and an additional partial ORF with weak similarities to S-layer proteins . The entire gene was sequenced to reveal a 1799-bp gene coding for 599 amino acids with a calculated molecular mass of 64.8 kDa . No transcription or translation signals could be determined in close proximity to the 5'-region . However, a strong putative terminator with a free energy of -16.84 kcal/mol was identified directly downstream of the gene . A PSI-Blast analysis showed similarities to members of S-layer proteins, cell-wall associated proteinases and hexosyl-transferases . Calculation of an unrooted phylogenetic tree with other examples of S-layer proteins and proteinases placed the deduced protein separately from both groups . A derivative of L . acidophilus NCFM was constructed by targeted integration into the gene . SDS-PAGE analysis of non-covalently linked proteins of the cell wall of the mutant, compared to the wild type, revealed the loss of a cell-surface protein . Phenotypic analyses of the mutant revealed significant changes in cell morphology, altered responses to various environmental stresses, and lowered cell adhesion . Based on the in silico and functional analyses, we ascertained that this protein plays a role in cell-wall processing during the growth and cell-cell separation and designated the gene as cell-division protein, cdpA.

Int J Food Microbiol, 2004 Dec 1, 97(1), 93 - 8
Development of culture media containing spent yeast cells of Debaryomyces hansenii and corn steep liquor for lactic acid production with Lactobacillus rhamnosus; Rivas B et al.; To develop a cost-effective fermentation medium, biomass of Debaryomyces hansenii coming from xylitol production and corn steep liquor was evaluated for lactic acid production by Lactobacillus rhamnosus in glucose-containing media . Glucose consumption and lactic acid generation were followed using a variety of media made with different nutrient supplementation . Under selected conditions, media containing glucose, biomass of D . hansenii and corn steep liquor as unique components led to product yields similar to those obtained in a fully supplemented medium.

Int J Food Microbiol, 2004 Dec 1, 97(1), 43 - 51
A collaborative study to evaluate qualitatively powdered baby food validation samples artificially contaminated with Salmonella anatum; Leuschner RG et al.; Nineteen laboratories participated in a collaborative study to evaluate the performance of validation samples consisting of powdered infant formula and cereal-based weaning food artificially contaminated with low levels of Salmonella anatum . The Standard method BS EN ISO 6579:2002 was to be followed for the trial . Salmonella counts in each baby food were around 10 CFU/25 g (low) or 10 CFU/g (high level) . Trial participants received five samples of each sample type and five blank samples . All samples contained Pseudomonas aeruginosa and Lactobacillus spp . at concentrations between 10(4) and 10(5) CFU/g as background flora . Precision data for the powdered infant formula were similar for the solid selective plating-out medium xylose-lysine-deoxycholate (XLD) and a 2nd choice of agar suitable to isolate Salmonella spp . The sensitivity was 91.3%, accordance 82.5% and concordance 83.9% for the low and 100% for the high level in all cases . For the cereal-based weaning food, the precision data for the high level were similar for XLD and the 2nd choice medium (98.8%, 97.5% and 97.5%) . A slight difference was observed for the low level where XLD obtained values of 82.5%, 73.8%, 70.6% for sensitivity, accordance and concordance and the second choice medium values of 81.5%, 72.5% and 69% . The specificity, accordance and concordance of blanks for the infant formula and cereal-based weaning food were 100%.

Med Dosw Mikrobiol, 2004, 56(1), 67 - 77
{Hydrogen peroxide produced by Lactobacillus species as a regulatory molecule for vaginal microflora}; Strus M et al.; A total of 33 strains of Lactobacillus belonging to 9 species, isolated from vagina, were tested for production of hydrogen peroxide . We observed that the following species: L . delbrueckii, L . acidophilus, L . crispatus, L . johnsonii and L . gasseri dominated over other species in secretion of hydrogen peroxide to the growth medium . Concentration of this substance amounted from 0.05 to 1.06 mM (in case of strong aeration the concentration increased up to 1.8 mM) . Moreover, killing properties of the pure hydrogen peroxide exerted toward Escherichia coli and Candida albicans were less prominent than these of the supernatants of cultures of Lactobacillus strains producing H2O2.

J Pediatr, 2004 Nov, 145(5), 612 - 6
Effect of probiotics on gastrointestinal symptoms and small intestinal permeability in children with atopic dermatitis; Rosenfeldt V et al.; OBJECTIVE: To determine whether probiotic lactobacilli may alleviate small intestinal inflammation and strengthen the intestinal barrier function in children with atopic dermatitis . STUDY DESIGN: In a double-blinded, placebo-controlled, cross-over study, probiotic lactobacilli (Lactobacillus rhamnosus 19070-2 and L reuteri DSM 12246) were administered for 6 weeks to 41 children with moderate and severe atopic dermatitis . Gastrointestinal symptoms were registered before and during treatment and small intestinal permeability was measured by the lactulose-mannitol test . RESULTS: During Lactobacillus supplementation, there was a significant decrease in the frequency of gastrointestinal symptoms (39% during the placebo period versus 10% during active treatment, P=.002) . There was a positive association between the lactulose to mannitol ratio and the severity of the eczema (r=0.61, P=.02 after placebo and r=0.53, P=.05 after active treatment) . After probiotic treatment, the lactulose to mannitol ratio was lower (0.073) than after placebo (0.110, P=.001) . CONCLUSIONS: Impairment of the intestinal mucosal barrier appears to be involved in the pathogenesis of atopic dermatitis . The study suggests that probiotic supplementation may stabilize the intestinal barrier function and decrease gastrointestinal symptoms in children with atopic dermatitis.

Plasmid, 2004 Nov, 52(3), 203 - 11
Complete nucleotide sequence and structural organization of pPB1, a small Lactobacillus plantarum cryptic plasmid that originated by modular exchange; de las Rivas B et al.; A small cryptic plasmid designated pPB1 was isolated from Lactobacillus plantarum BIFI-38 and its complete 2899 bp nucleotide sequence was determined . Sequence analysis revealed four putative open reading frames . Based on sequence analysis two modules could be identified . First, the replication module consisted of a sequence coding for a replication protein (RepB) and its corresponding target site, and two putative repressor proteins (RepA and RepC) . Sequence analysis indicated the possible synthesis of an antisense RNA that might regulate RepB production . A putative lagging-strand initiation site was also found, suggesting that pPB1 replicates via a rolling circle mechanism . The second module of pPB1 consisted of a sequence coding for a putative mobilization protein and its corresponding oriT site . Since the nucleotide sequence of the replication module showed 94.5% identity to the similar region on the Leuconostoc lactis plasmid pCI411, and the nucleotide sequence of the mobilization module had 97.5% identity to L . plantarum plasmid pLB4, it is concluded that pPB1 originated by modular exchange between two such plasmids by homologous recombination . Putative recombination sites where crossover might have taken place were also identified.

Obstet Gynecol, 2004 Nov, 104(5 Pt 1), 926 - 30
Vaginal yeast colonization in nonpregnant women: a longitudinal study; Beigi RH et al.; OBJECTIVE: We sought to investigate the prevalence and risk factors for vaginal yeast colonization over a 1-year period . METHODS: We conducted a longitudinal cohort study of 1,248 asymptomatic young women by collecting demographic and behavioral data at baseline, 4, 8, and 12 months . RESULTS: Seventy percent of women were colonized by vaginal yeast at one or more visits, but only 4% were colonized at all 4 visits . Using an adjusted generalized estimating equation model, factors associated with vaginal yeast colonization were marijuana use in the previous 4 months, depomedroxyprogesterone acetate use in the past 4 months, sexual intercourse in the previous 5 days, and concurrent colonization with lactobacilli and group B streptococcus . Symptoms of pruritus and vulvovaginal burning were associated with yeast colonization, but antifungal use was not . CONCLUSION: Recent sexual intercourse and use of injection contraceptives are risk factors for yeast colonization . Rates of antifungal use did not show an association with yeast colonization . The reporting of antifungal use by women lacking yeast colonization suggests that self-diagnosis is inaccurate . LEVEL OF EVIDENCE: II-2.

Mikrobiol Z, 2004 Jul-Aug, 66(4), 62 - 8
{Adhesion of lactic acid bacteria to epithelium of different cavities of human organism}
{Determination of species belonging of Lactobacillus genus strain with the use of rapd-typing}
{No authors listed}

The numeric analysis of 14 strains of lactobacilli investigated by 57 phenotypical characteristics, has resulted in formation of three clusters: group L . delbrueckii, L . plantarum and the modular group formed by strains of various species . The API-testing, 16S rRNA sequencing and RAPD-typing with primers 5'-AGCAGCGTGG-3' and LP3-ST 5'-TGGTCCGAGC-3' have been carried out for several strains which position in a dendrogram did not correspond to their species diagnosis . The obtained data have allowed the reclassification of the strain L . delbrueckii subsp . bulgaricus B-1923T as L . plantarum B-1923T, L . agilis B-2603 as L . fermentum B-2603, L . amylophilus 30/4/(2) as L . plantarum 30/4/(2) . Strain L . confusus 39T, identified by the phenotypical characteristics and data of API-testing as L . fermenturm, was reclassified on the basis of the phylogenetic analysis and RAPD-typing, as L . pentosus 39T.

J Obstet Gynaecol, 1998, 18(1), 63 - 7
A study of the prevalence of hydrogen peroxide generating Lactobacilli in bacterial vaginosis: the determination of H2O2 concentrations generated, in vitro , by isolated strains and the levels found in vaginal secretions of women with and without infection; Jones SA; The concentration of hydrogen peroxide in the vaginal secretions of normal women were compared with levels found in women infected with bacterial vaginosis (BV) . The mean concentration found in 90 normal samples was 0.171 mug per ml, whereas in women with BV, the mean level was 0.04 mug per ml ( P 0.001) . In vitro culture experiments using TMB agar showed that in normal women, 75% of Lactobacillus strains produced H O and 2 2 25% did not, whereas in women 'infected' with BV . 14% of the isolates produced H O and 86% did not ( P 0.001) . 2 2 Quantitative culture tests showed that both anaerobic and facultative strains of Lactobacilli isolated from normal women generated significantly higher levels of H O than 2 2 strains cultured from women with BV . Anaerobic strains isolated from normal women produced a mean level of 6.8 mug per ml, compared with the mean level of 1.4 mug per ml from women with BV . Facultative strains from normal women produced mean levels of 18.3 mug per ml compared with 2.4 mug per ml produced by strains from BV-infected women, which had been incubated under the same conditions ( P 0.001) . There results indicate that Lactobacilli which produce H O probably have a significant role in 2 2 protecting the vaginal ecosystem from BV infection.

Ann Clin Microbiol Antimicrob . 2004 Oct 27;3(1):23.
Effect of Ampicillin on the kinetics of colonization of Streptococcus pneumoniae and Lactobacillus fermentum in the respiratory tract of mice; Cangemi de Gutierrez R et al.; Ampicillin was selected to further study the effect of this antibiotic on the colonization capability of S . pneumoniae and L . fermentum intranasally inoculated in a mice experimental model . The sensitivity of S . pneumoniae and L . fermentum to antibiotics was evaluated by different "in vitro" techniques . The results showed that both microorganisms have a typical pattern of sensitivity to antibiotics in these assays . The "in vivo" experiments showed that the treatment with Ampicillin increased the number of lactobacilli and neumococci in the groups of mice treated only with one of the microorganisms . In those mice treated with Lactobacillus, challenged later with neumococci and treated with Ampicillin, the pathogen in lung decreased on the 4th day, disappearing completely after on . The histological studies showed that the antibiotic treatment decreased the inflammatory response produced by the pathogen at the lung and trachea levels.

J Food Prot, 2004 Oct, 67(10), 2306 - 9
Population dynamics of the constitutive biota of French dry sausages in a pilot-scale ripening chamber; Huerta R et al.; The population dynamic of constitutive biota on 84 samples belonging to two different types of French fermented dry sausages during the ripening process in a pilot-scale ripening chamber was investigated . Samples were analyzed in three steps of their production: fresh product, first drying stage, and finished product . In addition, 180 strains of lactic acid bacteria were identified using a miniaturized biochemical procedure of characterization . In general, the number of lactic acid bacteria that evolved during the ripening process of French dry sausages increased during the first days of the process after which the number of these organisms remained constant at approximately 8 log CFU/g . Lactobacillus sakei and Pediococcus pentosaceus, bacteria added as starter, were the dominant species . Pediococcus urinaeequi, Pediococcus acidilactici, and particularly Lactobacillus curvatus were also present . Finally, we have to take into account that the controlled conditions of the pilot plant generally contribute to the homogenization of the behavior of the starter biota.

J Food Prot, 2004 Oct, 67(10), 2189 - 94
Occurrence of Listeria monocytogenes in green table olives; Caggia C et al.; Microbiological safety of green table olives from different cultivars, prepared by both the Spanish-style and biological methods and fermented with starter cultures of lactic acid bacteria (Lactobacillus plantarum, Lactobacillus casei, and Lactobacillus pentosus), was investigated . The fermentation process was monitored by measuring pH values, titratable acidities, and growth of lactic acid bacteria over time . During fermentation, lactic acid bacteria and yeasts were major microbial populations . Microbiological safety was evaluated by analysis for Listeria monocytogenes with the use of an enrichment method during storage (from 55 days to 18 months) . Results demonstrated that L . monocytogenes can survive and grow in green table olives . L . monocytogenes was found in one of the commercial (thermally treated) samples analyzed and in all samples older than 2 months, irrespective of olive cultivar, lactic acid bacteria starter used, pH and titratable acidity of brine samples, or treatment applied.

Am J Obstet Gynecol, 2004 Oct, 191(4), 1324 - 30
Polymorphism in intron 2 of the interleukin-1 receptor antagonist gene, local midtrimester cytokine response to vaginal flora, and subsequent preterm birth; Genc MR et al.; OBJECTIVE: This study investigated the association between polymorphism in intron 2 of the interleukin-1 receptor antagonist gene, midtrimester vaginal microflora, vaginal interleukin receptor antagonist and interleukin-1beta levels and subsequent spontaneous preterm birth . STUDY DESIGN: Vaginal samples from 212 women, collected at 18-22 weeks' gestation, were analyzed for the polymorphism in intron 2 of the interleukin-1 receptor antagonist gene by polymerase chain reaction, qualitative and quantitative vaginal microflora, and interleukin-1beta and interleukin-1ra concentrations by enzyme-linked immunosorbent assay . Pregnancy outcome data were subsequently obtained . RESULTS: Carriage of intron 2 of the interleukin-1 receptor antagonist allele 2 (IL1RN * 2) was associated with an elevated vaginal pH in black ( P < .001) and white ( P = .005) women, a reduced interleukin-1beta response to anaerobic Gram-negative rods and/or Gardnerella vaginalis ( P < .01), and a decreased rate of spontaneous preterm deliveries (6% versus 18%, P = .02) . In black women, IL1RN * 2 carriage was associated with increased anaerobic Gram-negative rods, Mycoplasma, and Peptostreptococci and decreased Lactobacilli colonization . CONCLUSION: IL1RN * 2 carriage was associated with a blunted proinflammatory interleukin-1beta response to abnormal vaginal flora . This property may decrease susceptibility to infection-related preterm birth.

Swed Dent J, 2004, 28(3), 119 - 28
Comparing caries risk factors and risk profiles between children and elderly; Petersson GH et al.; The aim of this study was to compare the caries risk profiles of children and elderly, the actual annual caries increment and the impact of some selected caries related factors.The risk profiles were created by a computerised risk assessment program, the Cariogram, which evaluates data and presents the weighted and summarized result as one figure, illustrating the 'percent chance of avoiding caries' in the future . The data used originated from two separate longitudinal studies illustrating the Cariogram's capacity to assess caries risk . One study comprised about 400 children; the other included about 150 elderly . At baseline, information on past caries experience, diet, oral hygiene and use of fluoride was obtained . Saliva analyses included mutans streptococci and lactobacilli, buffering capacity and secretion rate . The caries risk was assessed and after two and five years, respectively, caries was re-evaluated and the incidence was compared with the predictions . Fifty percent of the children, but only two percent of the elderly appeared in the lowest caries risk group . Of the elderly, 26% belonged to the highest caries risk group versus 3 % of the children.The mean DFS increment per year for the total group of children was 0.4 +/- 0.8 (SD) and 1.2 +/- 1.9 for the elderly . Individual factors contributing significantly to the higher risk profiles for the adults were higher plaque scores, higher counts of mutans streptococci and lower buffering capacity . Over all, the risk for caries, as assessed by the Cariogram, was twice as high for the elderly.

Acta Crystallogr D Biol Crystallogr, 2004 Nov, 60(Pt 11), 2044 - 7 Epub 2004 Oct 20.
Crystallization and preliminary analysis of a water-forming NADH oxidase from Lactobacillus sanfranciscensis; Lountos GT et al.; Single crystals have been obtained of NADH oxidase (Nox), a flavoenzyme cloned from Lactobacillus sanfranciscensis . The enzyme catalyzes the oxidation of two equivalents of NAD(P)H and reduces one equivalent of oxygen to yield two equivalents of water, without releasing hydrogen peroxide after the reduction of the first equivalent of NAD(P)H . The enzyme crystallizes in space group P2(1)2(1)2(1), with unit-cell parameters a = 59.6, b = 92.6, c = 163.5 A . The crystals diffract to 1.85 A resolution using synchrotron radiation . Matthews coefficient calculations suggest the presence of two molecules per asymmetric unit (V(M) = 2.3 A(3) Da(-1), 45.5% solvent content), which has been confirmed by the molecular-replacement solution using a search molecule derived from NADH peroxidase (PDB code 1f8w).

Rocz Panstw Zakl Hig, 2004, 55(2), 139 - 45
{Microbiological quality of minimally processed vegetable salads}; Wojcik-Stopczynska B; The microbiological condition of minimally processed vegetable salads (7-days durability) purchased in the retail network of Szczecin has been assessed . The study included 14 kinds of salads manufactured by two producers . The total count of mesophilic aerobic bacteria and their spores, aid-forming bacteria (lactobacilli), titre of coliforms, occurrence of pathogenic bacteria and counts of moulds and yeast were determined . No pathogenic bacteria (Salmonella, Staphylococcus aureus, Bacillus cereus and Proteus) was detected in all salads . However contamination by saprophytic microorganisms was high in salads of both producers . Total count of mesophilic aerobic bacteria was higher than recommended level (> 10(5) cfu/g) in majority of salads . The high quantities of yeast (> 10(3) jtk/g) and lactobacilli (10(3)-10(5) fu/g) were also stated . The titre of coliforms was reduced (down to 0.01-0.0001 g) . The count of moulds did not exceed 100 cfu/g in a most of samples . Moulds occurring in tested samples were represented mainly by Penicillium sp.

Bioresour Technol, 2005 Mar, 96(4), 485 - 90
Selection of medium components by Plackett-Burman design for production of l(+) lactic acid by Lactobacillus amylophilus GV6 in SSF using wheat bran; Naveena BJ et al.; Plackett-Burman design was employed for screening 15 parameters for production of l(+) lactic acid from wheat bran, an inexpensive substrate and solid support, by Lactobacillus amylophilus GV6 in solid state fermentation (SSF) . Eleven nutrients belonging to two categories viz.; nitrogen sources and salt sources along with three physical parameters and a buffer were screened . This design screens {Formula: see text} variables in {Formula: see text} number of experiments . Coefficients and sum of squares ratio in percentage (SS%) of these variables were calculated by subjecting the experimental data to statistical analysis . The nitrogen sources peptone, yeast extract and tri-ammonium citrate, along with NaH(2)PO(4).2H(2)O and Tween 80, were found to influence productivity, which can be further optimized for increased lactic acid production . Use of this design is scarce in solid state fermentation and has not been attempted previously for single step conversion of starch to l(+) lactic acid using a bacterial system.

J Bacteriol, 2004 Nov, 186(21), 7069 - 83
Lactobacillus plantarum bacteriophage LP65: a new member of the SPO1-like genus of the family Myoviridae; Chibani-Chennoufi S et al.; The virulent Lactobacillus plantarum myophage LP65 was isolated from industrial meat fermentation . Tail contraction led to reorganization of the tail sheath and the baseplate; a tail tube was extruded . In ultrathin section the phage adsorbed via its baseplate to the exterior of the cell, while the tail tube tunneled through the thick bacterial cell wall . Convoluted membrane structures were induced in the infected cell . Progeny phage was detected 100 min postinfection, and lysis occurred after extensive digestion of the cell wall . Sequence analysis revealed a genome of 131,573 bp of nonredundant DNA . Four major genome regions and a large tRNA gene cluster were observed . One module corresponded to DNA replication genes . Helicase/primase and two replication/recombination enzymes represented the only links to T4-like Myoviridae from gram-negative bacteria . Another module corresponded to the structural genes . Sequence relatedness identified links with Listeria phage A511, Staphylococcus phage K, and Bacillus phage SPO1 . LP65 structural proteins were identified by two-dimensional proteome analysis and mass spectrometry . The putative tail sheath protein showed a shear-induced change in electrophoretic migration behavior . The genome organization of the structural module in LP65 resembled that of Siphoviridae from the lambda supergroup.

Curr Microbiol, 2004 Nov, 49(5), 341 - 5
Nutritional requirements of Lactobacillus delbrueckii subsp . lactis in a chemically defined medium; Hebert EM et al.; This study was undertaken to determine the nutritional requirements of Lactobacillus delbrueckii subsp . lactis and to develop a minimal chemically defined medium that supports sustained growth of these microorganisms . The single-omission technique was applied to each component of complete chemically defined medium in order to determine the nutritional requirements . L . delbrueckii subsp . lactis was prototrophic for alanine, glycine, aspartic acid, asparagine, glutamine, threonine, and proline . The lysine requirement was strain-dependent . Magnesium was the only essential oligoelement . These microorganisms also required uracil and guanine and adenine as pyrimidine and purine sources, respectively . In view of the nutritional requirements we designed a new minimal defined medium which supports sustained growth of L . delbrueckii subsp . lactis . This medium is simple and well defined, and should be preferable to complex media for conducting future biochemical, physiological, and genetic studies on L . delbrueckii subsp . lactis.

Lett Appl Microbiol, 2004, 39(5), 439 - 44
Screening of lactic acid bacteria from fermented vegetables by carbohydrate profiling and PCR-ELISA; Tamminen M et al.; AIMS: The aim of this study was to identify potential souring agents, isolated from fermented plant material, by API 50 CHL assay and a molecular method based on polymerase chain reaction and colorimetric hybridization (PCR-ELISA) . METHODS AND RESULTS: Forty-two strains of lactic acid bacteria derived from plant material were screened by taking advantage of API 50 CHL and PCR-ELISA . Oligonucleotide probes used for hybridization in PCR-ELISA were specific for lactobacilli, the Leuconostoc family, Lactobacillus pentosus/plantarum and Lactobacillus brevis . The hybrides were detected by a colour-developing reaction . Bacteria isolated from fermented cucumbers were identified as Lact . plantarum-related (Lact . plantarum and Lact . pentosus) and Leuconostoc species . Most of the strains isolated from sauerkraut were identified as Lact . pentosus/plantarum . CONCLUSIONS: Complementary results were obtained in the identification of bacterial strains, isolated from fermented cucumbers and sauerkraut, by API 50 CHL and PCR-ELISA . SIGNIFICANCE AND IMPACT OF THE STUDY: PCR-ELISA proved to be suitable for the screening of large numbers of bacterial isolates from fermented vegetables . This will be useful for the identification of strains suitable for the design of starter cultures for the fermentation of plant material.

Lett Appl Microbiol, 2004, 39(5), 431 - 8
Isolation and characterization of anti-Salmonella lactic acid bacteria from the porcine gastrointestinal tract; Casey PG et al.; AIMS: To identify lactic acid bacteria (LAB) of porcine intestinal origin with anti-Salmonella activity . METHODS AND RESULTS: Samples were obtained from pig faeces and caeca and screened for the presence of anti-Salmonella LAB . The 11 most promising isolates were identified as belonging to the genera Lactobacillus and Pediococcus . The LAB exhibited large variation in their ability to survive in simulated gastric juice at pH 1.85 . While Lactobacillus johnsonii species survived at levels of 80% for up to 30 min, Lactobacillus pentosus species declined to <0.001% in that time . All isolates tolerated porcine bile at a concentration of 0.3% (w/v), with some isolates capable of growth in the presence of up to 5% (w/v) bile . The ability of the LAB isolates to prevent Salmonella invasion of intestinal epithelial HT-29 cells varied, with reductions of between 30% (Lact . pentosus) and 80% (Lactobacillus murinus spp.) observed . CONCLUSIONS: LAB of porcine origin were observed to survive simulated passage through the GIT and inhibit growth of Salmonella and its invasion of the intestinal epithelium . SIGNIFICANCE AND IMPACT OF THE STUDY: The data demonstrate that some porcine intestinal LAB isolates may offer potential as probiotics for the reduction of Salmonella carriage in pigs.

Nutr Clin Care, 2004 Apr-Jun, 7(2), 69 - 74
The medicinal uses of poi; Brown AC et al.; Poi is a pasty starch made from the cooked, mashed corm of the taro plant (Colocasia esculenta L.) . Originating in Asia, this root crop is now found primarily in tropical and subtropical regions and was a major dietary staple in the Pacific islands . We hypothesize that poi has potential use as a probiotic-defined by FAO/WHO as, "live microorganisms which when administered in adequate amounts confer a health benefit to the host." No scientific studies have explored the possibility of poi being used as a probiotic in medical nutrition therapy, however, an investigator determined that the predominant bacteria in poi are Lactococcus lactis (95%) and Lactobacilli (5%), both of which are lactic acid-producing bacteria . This investigator also reported that poi contains significantly more of these bacteria per gram than yogurt . To determine if poi is beneficial for certain health conditions, a literature search was conducted to find all available research studies in which poi was used as a complementary treatment . Documented evidence suggests that poi shows promise for use in infants with allergies or failure-to-thrive . However, to support previous findings, more research needs to be conducted with poi and its potential use as a probiotic.

Appl Microbiol Biotechnol . 2004 Oct 9; {Epub ahead of print}
Development of an improved procedure for isolation and purification of exopolysaccharides produced by Lactobacillus delbrueckii subsp . bulgaricus NCFB 2483; Goh KK et al.; A method was developed for the isolation and purification of exopolysaccharide (EPS) produced by Lactobacillus delbrueckii subsp . bulgaricus NCFB 2483 that can be adapted for industrial-scale operation . Hydrolyzed milk medium, which was ultrafiltered to remove molecular species larger than 2.5x10(5) Da, was found to be a suitable growth medium for the bacteria, which produced approximately 400 mg EPS/l . Optimal isolation of EPS was achieved using centrifugation, filtration and ethanol precipitation methods . Insoluble and soluble EPS fractions were obtained . The soluble fraction was purified using a series of ethanol precipitations to achieve approximately 98% (w/w) purity . This fraction consisted of galactose, glucose, rhamnose and mannose in the ratio of approximately 5:1:0.6:0.5, with traces of glucosamine.

J Appl Microbiol, 2004, 97(5), 1095 - 104
Application of genotypic and phenotypic analyses to commercial probiotic strain identity and relatedness; Yeung PS et al.; AIMS: The objective of this study was to generate strain-specific genomic patterns of a bank of 67 commercial and reference probiotic strains, with a focus on probiotic lactobacilli . METHODS AND RESULTS: Pulsed-field gel electrophoresis (PFGE) was used as the primary method for strain differentiation . This method was compared with carbohydrate fermentation analysis . To supplement visual comparison, PFGE patterns were analysed quantitatively by cluster analysis using unweighted pair group method with arithmetic averages . SmaI, NotI and XbaI were found to effectively generate clear and easy-to-interpret PFGE patterns of a range of probiotic strains . Some probiotic strains from different sources shared highly similar PFGE patterns . CONCLUSIONS: Results document the value of genotypic strain identification methods, combined with phenotypic methods, for determining probiotic strain identity and relatedness . No correlation was found between relatedness determined by carbohydrate fermentation profiles alone compared with PFGE analysis alone . Some commercial strains are probably derived from similar sources . SIGNIFICANCE AND IMPACT OF THE STUDY: This approach is valuable to the probiotic industry to develop commercial strain identification patterns, to provide quality control of strain manufacturing production runs, to track use of protected strains and to determine the relatedness among different research and commercial probiotic strains.

J Agric Food Chem, 2004 Oct 20, 52(21), 6465 - 9
Metabolism of gallic acid and catechin by Lactobacillus hilgardii from wine; Alberto MR et al.; The ability of Lactobacillus hilgardii 5w to metabolize gallic acid and catechin was evaluated . It was grown in a complex medium containing gallic acid or catechin . The metabolites were analyzed by high-performance liquid chromatography and identified by comparing the retention times and spectral data with the standards of a database . In gallic acid-grown cultures, gallic acid, pyrogallol, catechol, protocatechuic acid, p-hydroxybenzoic acid, p-hydroxybenzaldehyde, and p-hydroxybenzyl alcohol were detected . In catechin-grown cultures, catechin, gallic acid, pyrogallol, catechol, p-hydroxybenzoic acid, acetovanillone, and homovanillic acid were detected . This work presents evidence of gallic acid and catechin degradation by L . hilgardii from wine .

FEMS Microbiol Lett, 2004 Oct 15, 239(2), 267 - 75
Rapid identification of probiotic Lactobacillus species by multiplex PCR using species-specific primers based on the region extending from 16S rRNA through 23S rRNA; Kwon HS et al.; This study aimed to develop a novel multiplex polymerase chain reaction (PCR) primer set for the identification of seven probiotic Lactobacillus species such as Lactobacillus acidophilus, Lactobacillus delbrueckii, Lactobacillus casei, Lactobacillus gasseri, Lactobacillus plantarum, Lactobacillus reuteri and Lactobacillus rhamnosus . The primer set, comprising of seven specific and two conserved primers, was derived from the integrated sequences of 16S and 23S rRNA genes and their rRNA intergenic spacer region of each species . It was able to identify the seven target species with 93.6% accuracy, which exceeds that of the general biochemical methods . The phylogenetic analyses, using 16S rDNA sequences of the probiotic isolates, also provided further support that the results from the multiplex PCR assay were trustworthy . Taken together, we suggest that the multiplex primer set is an efficient tool for simple, rapid and reliable identification of seven Lactobacillus species.

FEMS Microbiol Lett, 2004 Oct 15, 239(2), 213 - 20
Identification of the ornithine decarboxylase gene in the putrescine-producer Oenococcus oeni BIFI-83; Marcobal A et al.; We report here the identification of an ornithine decarboxylase (ODC) gene in the putrescine-producer Oenococcus oeni BIFI-83 strain . The gene contains a 2,235-nucleotide open reading frame encoding a 745-amino acid residues protein with a deduced molecular mass of 81 kDa . The primary structure of the ODC deduced from the nucleotide sequence has a consensus sequence containing the pyridoxal-5-phosphate (PLP) binding domain, and the critical amino acids residues involved in enzymatic activity are also conserved . As determined by BLAST analysis, the deduced amino acid sequence of the odc gene shares a 67% identity with the ODC protein from Lactobacillus 30a . The odc gene appears to be rarely present in the genome of O . oeni, since in a screening for the presence of this gene in 42 oenococcal strains none of the strains possessed an odc gene copy.

J Dent, 2004 Nov, 32(8), 667 - 78
In vitro growth, acidogenicity and cariogenicity of predominant human root caries flora; Shen S et al.; Streptococcus mutans (Sm), Lactobacillus acidophilus (La) and Actinomyces israelii (Ai) have been associated with root surface caries, which is an increasing problem in elderly Chinese . The aim of this study therefore, was to evaluate in vitro, the growth, acidogenicity and cariogenicity of these organisms, both in mono- and co-cultures using an in vitro model . Forty-eight root specimens were prepared using intact extracted human molars . Fresh, wild-type bacteria obtained from root caries lesions were assembled into seven experimental groups as either mono- or co-cultures and incubated with the root specimens . Appropriate controls were included . Growth curve of each experimental group was monitored for 24h, aerobically, at 37 degrees C using a microplate reader . The pH of the medium was recorded after 24-h incubation using a pH meter . Mean depths of artificial root lesions produced in each cultural group were measured using polarized light microscopy in specimens cut into thin sections (100+/-20 microm) . Compared with mono-cultures, synergistic growth was observed in co-cultures of 'La+Sm', 'Ai+La' and 'Ai+La+Sm' . Mean lesion depth produced in La group was significantly shallower than other mono- or co-culture groups (p<0.01) . The pH values of all culture media were similar after 24-h incubation . The current data elucidate the complex interactions of three predominant bacterial species considered prime agents of human root surface caries.

Eur J Obstet Gynecol Reprod Biol, 2004 Nov 10, 117(1), 76 - 81
Novel insight into the vaginal microflora in postmenopausal women under hormone replacement therapy as analyzed by PCR-denaturing gradient gel electrophoresis; Devillard E et al.; OBJECTIVES: A study was conducted to use new molecular technologies to identify the vaginal bacterial species of postmenopausal women under oral estrogen therapy (Premarin-conjugated equine estrogen, CEE) . STUDY DESIGN: Nineteen postmenopausal women under CEE treatment were recruited and their vaginal flora were analyzed during 3 months, using polymerase chain reaction in combination with denaturing gradient gel electrophoresis (DGGE) and sequencing of DNA fragments . RESULTS: Sixty-eight percent of the women presented with a 'Normal' Nugent score at the first sampling time (d0) . All the subjects had bacterial species detected in their vaginal flora . Lactobacillus species were detected in all samples . Moreover, 53% of the women had lactobacilli exclusively at d0 . Two Lactobacillus species were dominant, and were recovered from the majority of samples (L . iners and L . crispatus) . Forty-four percent of the samples also contained other bacterial species, which were potential urogenital pathogens . Candida albicans was detected in 26% of the subjects . The vaginal flora of the women under CEE treatment were different from that of postmenopausal women not receiving hormone therapy, but very similar to premenopausal women . CONCLUSIONS: The present study indicates that one benefit from hormone replacement therapy (HRT) is the restoration of a lactobacilli vaginal flora associated with a protective effect against urogenital infections.

Eur J Obstet Gynecol Reprod Biol, 2004 Nov 10, 117(1), 70 - 5
Efficacy and safety of vitamin C vaginal tablets in the treatment of non-specific vaginitis . A randomised, double blind, placebo-controlled study; Petersen EE et al.; METHODS: This was a randomised, double-blind, placebo-controlled study to evaluate the efficacy and safety of Vitamin C vaginal tablets (250 mg) given once a day in patients suffering from non-specific vaginitis . The total length of the study was 20 days, including a treatment phase of 6 days . The primary end-point was the presence in the two groups of non-specific vaginitis 1 and 2 weeks after the end of treatment, as assessed by at least 3 out of the 4 characteristic symptoms: discharge, fishy odour, vaginal pH >/= 4.7, and presence of clue cells . Secondary end-points were the individual symptoms and signs, above reported, and pruritus, fever, superinfections, microscopic findings on vaginal smear, and colposcopy . PATIENTS: One hundred female patients aged 18 years or older and suffering from non-specific vaginitis were included in the study after giving their informed consent . Fifty were randomised to the active treatment and 50 to placebo . Seven patients, three in the Vitamin C group and four in the placebo group, were lost to follow-up and did not complete the treatment period . Two patients in the active group showed protocol deviations (age under 18 years and HIV-positive, respectively) . The two groups resulted comparable for demographics, history and baseline clinical picture . RESULTS: A cluster analysis of the four main symptoms showed a statistically significant difference between the active group and the placebo group; significantly more patients were still affected by non-specific vaginitis after placebo (35.7%) compared to patients treated with Vitamin C tablets (14.0%) . The meaningful secondary variable, referring to the microscopic examination of vaginal smear, supported the trend for efficacy in the Vitamin C treated group . The clue cells disappeared in 79% of patients treated with the drug and in 53% of patients on placebo . Similarly, bacteria disappeared in 77 and 54%, respectively, while lactobacilli reappeared in 79.1 and 53.3%, respectively . Vaginal pH values decreased significantly in both groups, but the frequency rate of subjects with pH >/= 4.7, as measured 1 week after the drug discontinuation, was significantly lesser in the Vitamin C group (16.3%) than in the placebo group (38.6%) . Adverse events occurred in four patients, two on placebo (pruritus, cystitis) and two on Vitamin C (two candidiasis).

Clin Infect Dis, 2004 Oct 1, 39(7), 990 - 5 Epub 2004 Sep 02.
Bacterial vaginosis and anaerobic bacteria are associated with endometritis; Haggerty CL et al.; BACKGROUND: Chlamydia trachomatis and/or Neisseria gonorrhoeae account for approximately one-third to one-half of pelvic inflammatory disease (PID) cases . Thus, up to 70% of cases have an unknown, nongonococcal/nonchlamydial microbial etiology . METHODS: We investigated the associations of N . gonorrhoeae, C . trachomatis, bacterial vaginosis, anaerobic bacteria, facultative bacteria, and lactobacilli with endometritis among 278 women with complete endometrial histology and culture from the PID Evaluation and Clinical Health Study . RESULTS: Women with acute endometritis were less likely to have H(2)O(2)-producing Lactobacillus species (odds ratio {OR}, 0.1; 95% confidence interval {CI}, 0.01-0.8) and more likely to be infected with C . trachomatis (OR, 16.2; 95% CI, 4.6-56.6), N . gonorrhoeae (OR, 11.6; 95% CI, 4.5-29.9), diphtheroids (OR, 5.0; 95% CI, 2.1-12.2), black-pigmented gram-negative rods (OR, 3.1; 95% CI, 1.4-7.0), and anaerobic gram-positive cocci (OR, 2.1; 95% CI, 1.0-4.3) and to have bacterial vaginosis (OR, 2.4; 95% CI, 1.3-4.3) . CONCLUSIONS: We conclude that bacterial vaginosis-associated organisms are frequent among women with PID . Because these organisms were strongly associated with endometritis, we recommend that all women with PID be treated with regimens that include metronidazole.

Biotechnol Bioeng, 2004 Dec 5, 88(5), 601 - 12
A method for the determination of flux in elementary modes, and its application to Lactobacillus rhamnosus; Poolman MG et al.; In this article we address the question of how, given information about the reaction fluxes of a system, flux values can be assigned to the elementary modes of that system . Having described a method by which this may be accomplished, we first illustrate its application to a hypothetical, in silico system, and then apply it to fermentation data from Lactobacillus rhamnosus . This reveals substantial changes in the flux values assigned to elementary modes, and thus to the internal metabolism, as the fermentation progresses . This is information that could not, to our knowledge, be obtained by existing methods . The relationship between our technique and the well-known method of Metabolic Flux Analysis is also discussed.

Br J Nutr, 2004 Sep, 92(3), 439 - 46
The in vivo use of the stable isotope-labelled biomarkers lactose-{15N}ureide and {2H4}tyrosine to assess the effects of pro- and prebiotics on the intestinal flora of healthy human volunteers; De Preter V et al.; Amongst the various claimed beneficial effects of pro- and prebiotics for the human host, it has been hypothesised that functional foods are able to suppress the generation and accumulation of toxic fermentation metabolites (NH3, p-cresol) . Direct evidence supporting this hypothesis is lacking mainly because of the unavailability of reliable biomarkers . Preliminary data indicate that lactose-{15N}ureide and {2H4}tyrosine may be potential biomarker candidates . The aim of the present study was to evaluate the effect of pro- and prebiotics on the colonic fate of these biomarkers in a randomised, placebo-controlled, cross-over study with nineteen healthy volunteers . At the start of the study and at the end of each 2-week study period, during which they were administered either a probiotic (n 10; 6.5 x 10(9) Lactobacillus casei Shirota cells twice daily) or a prebiotic (n 9; lactulose 10 g twice daily), the volunteers consumed a test meal containing the two biomarkers . Urine was collected during 48 h . Results were expressed as percentage of the administered dose . As compared with the placebo, the decrease in the percentage dose of p-{2H4}cresol in the 24-48 h urine fraction was significantly higher after probiotic intake (P=0.042) . Similar changes were observed for the 15N tracer (P=0.016) . After prebiotic intake, a significantly higher decrease in the percentage dose of p-{2H4}cresol (P=0.005) and 15N tracer (P=0.029) was found in the 0-24 h urine collection . The present results demonstrate that suppression of the generation and accumulation of potentially toxic fermentation metabolites by pro- and prebiotics can reliably be monitored in vivo by the use of stable isotope-labelled biomarkers.

Int Arch Allergy Immunol, 2004 Nov, 135(3), 205 - 15 Epub 2004 Nov.
The anti-allergic effects of lactic acid bacteria are strain dependent and mediated by effects on both Th1/Th2 cytokine expression and balance; Fujiwara D et al.; BACKGROUND: There is growing interest in the immune-stimulating effect and in particular, the anti-allergic effect, of lactic acid bacteria (LAB) . However, no comprehensive studies have been done that compare the immune-stimulating potential of LAB strains . METHODS: The in vitro immune-stimulating effects on Th1/Th2 balance of more than 100 LAB strains were compared in splenocytes from ovalbumin-sensitized Th2-polarized mice . The in vivo anti-allergic ability of strain KW3110 was studied in the Th2-polarized model by detecting serum IgE concentration, Th1/Th2 cytokine secretion from splenocytes, and the expression of co-stimulatory molecules on macrophages . RESULTS: In vitro studies from Th2-polarized splenocytes, using IL-12 as a Th1 parameter and IL-4 secretion as a Th2 parameter revealed a wide variety of IL-12-inducing and IL-4-repressing activities, depending on the strain of LAB, not depending on the species . However, evaluation of individual strains in vivo revealed that after exposure to Lactobacillus paracasei KW3110 strain, the serum IgE elevation elicited by repeated OVA injection of mice was strongly inhibited . Cytokine secretion from splenocytes 20 weeks after KW3110 administration showed increased IL-12 and decreased IL-4 expression . Both CD40 and B7-1 expression on macrophages was upregulated by administration of KW3110 . CONCLUSIONS: Improving the consequences of the Th1/Th2 imbalance by administration of LAB was dependent upon the LAB strain rather than the LAB species . Oral KW3110 administration in the mouse allergy model directed the Th1/Th2 balance toward Th1 through the maturation of APCs and inhibition of serum IgE elevation.

Appl Environ Microbiol, 2004 Oct, 70(10), 6290 - 5
Molecular characterization of two lactate dehydrogenase genes with a novel structural organization on the genome of Lactobacillus sp . strain MONT4; Weekes J et al.; Two lactate dehydrogenase (ldh) genes from Lactobacillus sp . strain MONT4 were cloned by complementation in Escherichia coli DC1368 (ldh pfl) and were sequenced . The sequence analysis revealed a novel genomic organization of the ldh genes . Subcloning of the individual ldh genes and their Northern blot analyses indicated that the genes are monocistronic.

Appl Environ Microbiol, 2004 Oct, 70(10), 5929 - 36
Improved stress tolerance of GroESL-overproducing Lactococcus lactis and probiotic Lactobacillus paracasei NFBC 338; Desmond C et al.; The bacterial heat shock response is characterized by the elevated expression of a number of chaperone complexes . Two-dimensional polyacrylamide gel electrophoresis revealed that GroEL expression in probiotic Lactobacillus paracasei NFBC 338 was increased under heat adaptation conditions (52 degrees C for 15 min) . Subsequently, the groESL operon of L . paracasei NFBC 338 was PCR amplified, and by using the nisin-inducible expression system, two plasmids, pGRO1 and pGRO2, were constructed on the basis of vectors pNZ8048 and pMSP3535, respectively . These vectors were transferred into Lactococcus lactis(pGRO1) and L . paracasei(pGRO2), and after induction with nisin, overexpressed GroEL represented 15 and 20% of the total cellular protein in each strain, respectively . Following heat shock treatment of lactococci (at 54 degrees C) and lactobacilli (at 60 degrees C), the heat-adapted cultures maintained the highest level of viability (5-log-unit increase, approximately) in each case, while it was found that the GroESL-overproducing strains performed only moderately better (1-log-unit increase) than the controls . On the other hand, the salt tolerance of both GroESL-overproducing strains (in 5 M NaCl) was similar to that of the parent cultures . Interestingly, both strains overproducing GroESL exhibited increased solvent tolerance, most notably, the ability to grow in the presence of butanol (0.5% {vol/vol}) for 5 h, while the viability of the parent strain declined . These results confirm the integral role of GroESL in solvent tolerance, and to a lesser extent, thermotolerance of lactic acid bacteria . Furthermore, this study demonstrates that technologically sensitive cultures, including certain probiotic lactobacilli, can potentially be manipulated to become more robust for survival under harsh conditions, such as food product development and gastrointestinal transit.

Appl Environ Microbiol, 2004 Oct, 70(10), 5750 - 5
Mechanisms of bactericidal action of cinnamaldehyde against Listeria monocytogenes and of eugenol against L . monocytogenes and Lactobacillus sakei; Gill AO et al.; The spice oil components eugenol and cinnamaldehyde possess activity against both gram-positive and gram-negative bacteria, but the mechanisms of action remain obscure . In broth media at 20 degrees C, 5 mM eugenol or 30 mM cinnamaldehyde was bactericidal (>1-log reduction in the number of CFU per milliliter in 1 h) to Listeria monocytogenes . At a concentration of 6 mM eugenol was bactericidal to Lactobacillus sakei, but treatment with 0.5 M cinnamaldehyde had no significant effect . To investigate the role of interference with energy generation in the mechanism of action, the cellular and extracellular ATP levels of cells in HEPES buffer at 20 degrees C were measured . Treatment of nonenergized L . monocytogenes with 5 mM eugenol, 40 mM cinnamaldehyde, or 10 microM carbonyl cyanide m-chlorophenylhydrazone (CCCP) for 5 min prevented an increase in the cellular ATP concentration upon addition of glucose . Treatment of energized L . monocytogenes with 40 mM cinnamaldehyde or 10 microM CCCP caused a rapid decline in cellular ATP levels, but 5 mM eugenol had no effect on cellular ATP . Treatment of L . sakei with 10 mM eugenol prevented ATP generation by nonenergized cells and had no effect on the cellular ATP of energized cells . CCCP at a concentration of 100 microM had no significant effect on the cellular ATP of L . sakei . No significant changes in extracellular ATP were observed . Due to their rapidity, effects on energy generation clearly play a major role in the activity of eugenol and cinnamaldehyde at bactericidal concentrations . The possible mechanisms of inhibition of energy generation are inhibition of glucose uptake or utilization of glucose and effects on membrane permeability.

Ren Fail, 2004 Jul, 26(4), 419 - 23
Lactobacillus peritonitis: a rare cause of peritonitis in peritoneal dialysis patients; Lee SY et al.; Lactobacilli are part of the normal gastrointestinal and female genitourinary flora in humans and they are seldom pathogenic and rarely cause human disease . In the literature, Lactobacillus peritonitis was most common in immunocompromised patients, including patients under chronic peritoneal dialysis . We also suspect that the presence of Lactobacillus spp . in the peritoneal fluid might indicate the leakage of normal flora from a perforated intraabdominal hollow organ . To access the versatile clinical pictures of Lactobacillus peritonitis, this investigation retrospectively reviewed the medical records for Lactobacillus spp . isolated from peritoneal fluid from July 1998 to January 2002 at Chang-Gung Memorial Hospital, Taipei, Taiwan . A total of 10 patients were enrolled in the study . Six of these 10 patients had concomitant intraabdominal hollow organ perforation, and peritoneal fluid cultures in these six patients also contained bacteria other than Lactobacillus spp . All six patients had recently experienced either abdominal surgery or blunt abdominal trauma . The remaining four patients who had not undergone surgery had decompensated liver cirrhosis with ascites and spontaneous bacterial peritonitis . The results suggested that the presence of Lactobacillus spp . in the peritoneal fluid other than immunocompromised patients should raise the suspicion of hollow organ perforation in patients with recent abdominal surgery or blunt abdominal trauma.

Infect Dis Obstet Gynecol, 2004, 12(1), 13 - 6
Cytolytic vaginosis: misdiagnosed as candidal vaginitis; Cerikcioglu N et al.; OBJECTIVES: In this study, 210 women with vaginal discharge and other symptoms/signs of genital pathology suggestive of vulvovaginal candidiasis (VVC) were involved in order to distinguish true WC and cytolytic vaginosis (CV) cases . METHODS: Fungal cultures, 10% potassium hydroxide (KOH) and Gram stained preparations and pH measurements were performed on the vaginal discharge material of each patient . RESULTS: Fifteen patients (7.1%) were diagnosed with cytolytic vaginosis according to their clinical and microbiological findings, including abundant lactobacilli, fragmented epithelial cells and/or free nuclei due to cytolysis, seen in their discharge materials on microscopic examination, but no fungal growth . CONCLUSIONS: The results of this study may contribute to the reports in the literature indicating the importance of such disorders, which are generally misdiagnosed as candidiasis.

Antibiot Khimioter, 2004, 49(4), 35 - 8
{Large intestine dysbacteriosis and therapy efficacy in patients with respiratory tract tuberculosis in sanatoria}; Accumulation of Mn(II) in Deinococcus radiodurans facilitates gamma-radiation resistance; Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USA . mdaly@usuhs.mil

Deinococcus radiodurans is extremely resistant to ionizing radiation . How this bacterium can grow under chronic gamma radiation {50 grays (Gy) per hour} or recover from acute doses greater than 10 kGy is unknown . We show that D . radiodurans accumulates very high intracellular manganese and low iron levels compared with radiation-sensitive bacteria and that resistance exhibits a concentration-dependent response to manganous chloride {Mn(II)} . Among the most radiation-resistant bacterial groups reported, Deinococcus, Enterococcus, Lactobacillus, and cyanobacteria accumulate Mn(II) . In contrast, Shewanella oneidensis and Pseudomonas putida have high iron but low intracellular manganese concentrations and are very sensitive . We propose that Mn(II) accumulation facilitates recovery from radiation injury.

Obstet Gynecol, 2004 Oct, 104(4), 761 - 9
Bacterial vaginosis and risk of pelvic inflammatory disease; Ness RB et al.; BACKGROUND: Bacterial vaginosis commonly is found in women with pelvic inflammatory disease (PID), but it is unclear whether bacterial vaginosis leads to incident PID . METHODS: Women (n = 1,179) from 5 U.S . centers were evaluated for a median of 3 years . Every 6-12 months, vaginal swabs were obtained for gram stain and culture of microflora . A vaginal microflora gram stain score of 7-10 was categorized as bacterial vaginosis . Pelvic inflammatory disease was diagnosed by presence of either histologic endometritis or pelvic pain and tenderness plus one of the following: oral temperature greater than 38.3 degrees C; sedimentation rate greater than 15 mm/hour; white blood count greater than 10,000; or lower genital tract detection of leukorrhea, mucopus, or Neisseria gonorrhoeae or Chlamydia trachomatis . RESULTS: After adjustment for relevant demographic and lifestyle factors, baseline bacterial vaginosis was not associated with the development of PID (adjusted hazard ratio 0.89, 95% confidence interval 0.55-1.45) . Carriage of bacterial vaginosis in the previous 6 months before a diagnosis (adjusted risk ratio 1.31, 95% confidence interval 0.71-2.42) also was not significantly associated with PID . Similarly, neither absence of hydrogen peroxide-producing Lactobacillus nor high levels of Gardnerella vaginalis significantly increased the risk of PID . Dense growth of pigmented, anaerobic gram-negative rods in the 6 months before diagnosis did significantly increase a woman's risk of PID (P =.04) . One subgroup of women, women with 2 or more recent sexual partners, demonstrated associations among bacterial vaginosis, Gardnerella vaginalis, anaerobic gram-negative rods, and PID . CONCLUSION: In this cohort of high-risk women, after adjustment for confounding factors, we found no overall increased risk of developing incident PID among women with bacterial vaginosis . LEVEL OF EVIDENCE: II-2

Eur J Oral Sci, 2004 Oct, 112(5), 424 - 8
Protective effect of salivary nitrate and microbial nitrate reductase activity against caries; Doel JJ et al.; To test the hypothesis that a combination of high salivary nitrate and high nitrate-reducing capacity are protective against dental caries, 209 children attending the Dental Institute, Barts and The London NHS Trust were examined . Salivary nitrate and nitrite levels, counts of Streptococcus mutans and Lactobacillus spp., and caries experience were recorded . Compared with control subjects, a significant reduction in caries experience was found in patients with high salivary nitrate and high nitrate-reducing ability . Production of nitrite from salivary nitrate by commensal nitrate-reducing bacteria may limit the growth of cariogenic bacteria as a result of the production of antimicrobial oxides of nitrogen, including nitric oxide.

J Physiol Biochem, 2004 Jun, 60(2), 85 - 91
Daily ingestion of fermented milk containing Lactobacillus casei DN114001 improves innate-defense capacity in healthy middle-aged people; Parra MD et al.; Different lactic acid bacteria have often been administered as a dietary means to enhance immune system activity . Based on this statement, the aim of the current work was to test the effects of a Lactobacillus casei DN114001 fermented milk consumption on the immune response capacity in middle-age volunteers . Forty-five healthy volunteers, 24 women and 21 men (aged: 51-58 years), were randomized into two groups to receive three cups per day of a L . casei DN114001 (10(8)-10(10) ufc/g) fermented milk (n = 23), or placebo (n = 22), during an 8-week period . Measurements were performed before (day 0), and after the nutritional intervention (day 56) . After the trial, no changes in immune cell proportions were detected, but the probiotic-treated group increased oxidative burst capacity of monocytes (probiotic group: p = 0.029; placebo group: p = 0.625), as well as NK cells tumoricidal activity (probiotic group: p = 0.023; placebo group: p = 0.125) . Results showed that daily intake of fermented milk containing Lactobacillus casei DN114001 could have a positive effect in modulating the innate immune defense in healthy-middle-age people.

Rev Iberoam Micol, 2003 Dec, 20(4), 160 - 3
{In vitro effect of carbohydrates and enteric bacteria on adherence of Candida albicans}; Biasoli MS et al.; The adherence of Candida albicans to any cell is considered essential in the process that leads to colonization . Our objective in this study was to evaluate the effect of different carbohydrates and the presence of lactobacilli and Escherichia coli on the in vitro adherence of Candida albicans . The adherence to buccal epithelial cells was higher when growing at concentrations of galactose of 50, and 200 mM, as well as 50, 200, and 500 mM of sucrose, and 500 mM of mannose, compared with that obtained when growing in Sabouraud dextrose broth (p < 0.01) . The presence of other microorganisms, such as Lactobacillus acidophilus and L . casei, caused a decrease in the in vitro adherence of C . albicans to buccal epithelial cells (p < 0.05), whereas E . coli did not modify this adherence at all.

Mikrobiol Z, 2004 May-Jun, 66(3), 33 - 42
{Screening of the strains of lactic acid bacteria possessing hypocholesterinemic activity and their practical use}; Stabilization of cucurbitacin E-glycoside et al.; USDA/ARS Insect Biocontrol Laboratory, Beltsville, MD 20705, USA . pmartin@asrr.arsusda.gov

Cucurbitacins are feeding stimulants for diabroticite beetles, including corn rootworms and cucumber beetles, which can be added to a bait containing an insecticide thereby reducing the levels of other insecticide treatments needed to control these pests . One of them, cucurbitacin E-glycoside, is water soluble and easily processed from mutant bitter Hawkesbury watermelons (BHW) that express elevated levels of cucurbitacin . Storage of BHW extract at room temperature resulted in a 92% reduction of cucurbitacin E-glycoside over two months, while refrigeration or freezing resulted in a 60% loss of the active ingredient during this time . The loss of the active ingredient was correlated with an increase in BHW extract pH from 5 to greater than 9 . The increase in pH of the BHW extracts at room temperature appeared to be due to the growth of certain bacteria, especially Bacillusspp . In refrigerated extracts, the pH remained relatively constant, and bacterial growth was dominated by bacteria such as Lactobacilli . An alternative to refrigeration is concentration of BHW extract . One means of concentration is spray drying, but the high sugar content of the BHW extract (20mg/ml glucose, 40mg/ml fructose) makes this technique impractical . Fermentation of the BHW extract by the yeast, Saccharomyces boulardii, eliminated the sugars and did not raise the pH nor alter the cucurbitacin E-glycoside content of the extract . Elimination of the sugars by fermentation produced an extract that could be successfully spray dried . BHW extract fermented by S . boulardii produced a higher level of feeding stimulation for spotted cucumber beetles in laboratory choice tests . When applied to cucumbers, there was no difference in control of spotted and striped cucumber beetles between baits of fresh or fermented juices combined with the same insecticide.

Int J Food Microbiol, 2004 Nov 15, 96(3), 307 - 13
Evidence on the role of protein biosynthesis in the induction of heat tolerance of Lactobacillus rhamnosus GG by pressure pre-treatment; Ananta E et al.; It was the aim of this work to evaluate, whether and to which extent heat resistance of Lactobacillus rhamnosus GG is affected by mild pressure treatments prior to exposure to lethal temperatures, such as during spray-drying . It was observed that cells pressure pre-treated at 100 MPa at 37 degrees C for 10 min showed higher survival than untreated cells when exposed to heat challenge at 60 degrees C . To gain more insights on the cellular mode of action of pressure induced heat tolerance, flow cytometric analysis was applied in combination with functional dye LIVE/DEAD BacLight bacterial viability kit . Dot plot analysis showed that a lower degree of membrane damage was observed at pressure pre-treated cells upon heat treatment at 60 degrees C for 3 min . Evaluation of heat inactivation kinetics of cells pressure treated in the presence of chloramphenicol, a protein synthesis inhibitor, pointed out the potential contribution of pressure-induced protein biosynthesis in the enhancement of bacterial heat tolerance.

Int J Food Microbiol, 2004 Nov 15, 96(3), 273 - 9
Characterization of lytic enzyme activities of Lactobacillus gasseri with special reference to autolysis; Yokoi KJ et al.; Lactobacillus gasseri JCM 1130 and JCM 1131(T) exhibited autolytic activity in agar containing autoclaved cells of each strain as substrate . By zymogram analysis of JCM 1131(T), two lytic bands with apparent molecular masses of 54.5 and 35 kDa, were detected . Similarly, JCM 1130 yielded two lytic bands with apparent molecular masses of 35 and 33.5 kDa . In simple buffers as well, JCM 1131(T) suffered a drastic decrease in cell turbidity, but JCM 1130 did not undergo the decrease . The optimal pH for autolysis of JCM 1131(T) was in the range of 6.0-7.0, and the lysis was completely inhibited at pH 4-5 . The lysis of JCM 1131(T) was suppressed by NaCl, in a concentration-dependent way . When subjected to UV irradiation or mitomycin C (MMC) treatment, cultures of both strains elicited conspicuous turbidity decrease after 2-4 h of growth, suggesting the occurrence of prophage induction . The 35-kDa lytic band of JCM 1131(T) and the 33.5-kDa protein of JCM 1130 were considerably increased by UV irradiation.

J Agric Food Chem, 2004 Oct 6, 52(20), 6300 - 5
Phytate degradation by lactic acid bacteria and yeasts during the wholemeal dough fermentation: a 31P NMR study; Reale A et al.; myo-Inositol hexaphosphate (IP6) is the main source of phosphorus in cereal grains, and therefore, in bakery products . Different microorganisms such as yeasts and lactic acid bacteria have phytase enzymes able to hydrolyze IP6 during the wholemeal breadmaking . In this paper, the phytase activity of Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus curvatus, and Saccharomyces cerevisiae strains, isolated from southern Italian sourdoughs, is assayed using the (31)P NMR technique . The sourdough technology based on the use of lactic acid bacteria in the breadmaking is finally suggested.

J Dairy Sci, 2004 Jun, 87(6), 1585 - 94
Impact of autolytic, proteolytic, and nisin-producing adjunct cultures on biochemical and textural properties of cheddar cheese; Sallami L et al.; The effect of incorporating a highly autolytic strain (Lactobacillus delbrueckii subsp . bulgaricus UL12) a proteolytic strain (Lactobacillus casei subsp . casei L2A), or a nisin Z-producing strain (Lactococcus lactis, subsp . lactis biovar diacetylactis UL719) into Cheddar cheese starter culture (Lactococcus lactis KB and Lactococcus cremoris KB) on physicochemical and rheological properties of the resultant cheeses was examined . Cheeses were ripened at 7 degrees C and analyzed over a 6-mo period for viable lactococcal and lactobacilli counts, pH, titratable acidity (TA), lipolysis, proteolysis, and textural characteristics . The combination of the nisin-producing strain and autolytic adjuncts significantly increased the production of water-soluble nitrogen, free amino acids, and free fatty acids . The effect of Lc . diacetylactis UL719 alone or of Lb . casei L2A on water-soluble nitrogen and free amino acid contents were also significant, whereas their effect on free fatty acids was not . Viable counts of Lb . bulgaricus UL12 were significantly reduced in the presence of Lc . diacetylactis UL719 . Lactobacilli-containing cheeses showed significantly lower values for hardness, fracturability, and springiness . It could be concluded that the addition of Lb . bulgaricus UL12 together with a nisin-producing strain produces a greater increase in cheese proteolysis and an improvement in Cheddar cheese texture.

FEMS Microbiol Lett, 2004 Oct 1, 239(1), 111 - 6
Evaluation of the robustness of FT-IR spectra of lactobacilli towards changes in the bacterial growth conditions; Oust A et al.; In order to use Fourier Transform Infrared (FT-IR) spectroscopy for identification of microorganisms on a routine basis, it is important that the spectra are robust against small, uncontrollable variations in the bacterial growth conditions . In this study, the effect of small variations in growth temperature, growth time, growth medium and atmospheric conditions on the separation of Lactobacillus based on their FT-IR spectra was investigated . The resulting spectra were shown to be robust against the variations in the cultivation conditions, and the separation of both strains and species was unaffected . Larger variations in the growth medium influenced only the separation of strains . FT-IR spectroscopy for identification of lactobacilli therefore seems to be robust against small variations in the cultivation conditions.

Ultramicroscopy, 2004 Nov, 101(2-4), 105 - 9
Nanosurgery: observation of peptidoglycan strands in Lactobacillus helveticus cell walls; Firtel M et al.; The internal cell wall structure of the bacterium Lactobacillus helveticus has been observed in situ in aqueous solution using an atomic force microscope (AFM) . The AFM tip was used not only for imaging but presumably to remove mechanically large patches of the outer cell wall after appropriate chemical treatment, which typically leaves the bacteria alive . The surface exposed after this "surgery" revealed approximately 26 nm thick twisted strands within the cell wall . The structure and location of the observed strands are consistent with the glycan backbone of peptidoglycan fibers that give strength to the cell wall . The structural organization of these fibers has not been observed previously.

JPEN J Parenter Enteral Nutr, 2004 Sep-Oct, 28(5), 348 - 54
Effects of a nutritional supplement on the immune response and cytokine production in free-living Chilean elderly; Bunout D et al.; BACKGROUND: Immune response is impaired in the elderly . Our aim was to study the effects of a special nutritional formula on the immune response and response to influenza and pneumococcal vaccination in elderly subjects . METHODS: Sixty healthy subjects aged > or = 70 years, with a Mini Mental score > or = 22 were studied . Half of the subjects received a special nutritional formula (in addition to the regular diet) providing, among other nutrients, 480 kcal, 31 g proteins, 120 IU vitamin E, 3.8 microg vitamin B12, 400 microg folic acid, 10(9) cfu Lactobacillus paracasei (NCC 2461), and 6 g of fructo-oligosaccharides . At 4 months of follow-up, subjects were vaccinated against influenza and pneumococcus . Lymphokine production by mononuclear cells (PBMC), lymphocyte subpopulations, and natural killer cell (NK) activity were measured at baseline and 4 months of follow-up (before vaccination) . Antibodies against influenza and pneumococcal antigens and flu-stimulated production of interferon gamma and interleukin-2 by PBMC were measured at 4 and 6 months . Skin response to 7 recall antigens and body composition were assessed at baseline and at 4 and 12 months . All infections occurring during the study period were recorded . RESULTS: NK activity increased in supplemented subjects and decreased in nonsupplemented individuals . Interleukin-2 production by PBMC and the proportion of T cells with NK activity decreased in controls and did not change in supplemented subjects . Supplemented subjects reported less infections than nonsupplemented individuals (in 13% and 22% of scheduled visits, respectively; p = .02) . CONCLUSIONS: This nutritional supplement increased innate immunity and protection against infections in elderly people.

Arch Microbiol, 2004 Dec, 182(6), 450 - 457 Epub 2004 Sep 23.
Identification of a region involved in the pheromone receptor function of the histidine kinase PlnB; Johnsborg O et al.; Bacteriocin biosynthesis in Lactobacillus plantarum is an inducible process, triggered by the secreted inducer peptide pheromone IP-C11 . The environmental concentration of IP-C11 is monitored by the membrane-bound histidine protein kinase PlnB, which is part of a two-component signal transduction pathway . Upon interaction with IP-C11, PlnB phosphorylates the cognate response regulator PlnC . This regulator subsequently activates transcription of the bacteriocin genes . PlnB belongs to the HPK(10) subfamily of peptide-pheromone-activated histidine kinases . All members of this subfamily have an unusual polytopic membrane domain that previously has been shown to contain the peptide pheromone receptor . Employing an in vivo reporter assay, the present work investigated the receptor functionality of various mutagenized PlnB membrane domains . The results indicated that important determinants for receptor function locate to the most N-terminal extracytoplasmatic loop of the membrane domain . In addition, this region appears to be involved in the peptide pheromone interaction of ComD, another member of the HPK(10) subfamily.

Biotechnol Bioeng, 2004 Oct 5, 88(1), 71 - 6
New drying process for lactic bacteria based on their dehydration behavior in liquid medium; Mille Y et al.; This study describes the different stages of optimization in an original drying process for lactic acid bacteria that allows the retrieval of dried samples of Lactobacillus plantarum with maximum viability . The process involves the addition of casein powder to bacterial pellets, followed by mixing and then air-drying in a fluidized bed dryer . The effects on bacterial viability of the a(w) of the casein powder and the kinetics of a(w) variation in the fluidized bed dryer are considered . These parameters were first studied in a water-glycerol solution and the results were then applied to the drying process . Data from the study in liquid medium were reliable in the fluidized drying stage, insofar as optimal viability was achieved for similar dehydration times (16-50 min in liquid medium, and 30 min in the fluidized bed dryer) . However, when the powder was mixed rapidly with bacteria, the level of destruction differed from that observed in liquid medium . Viability was up to 70% when the a(w) of water-glycerol was 0.55, whereas it was only 2.1% when the a(w) of the casein-bacterial mix was 0.64 . The predictive capacity of dehydration in liquid medium is discussed with regard to the permeability of cells to external solutes . The new process allowed 100% survival of L . plantarum after complete drying (final a(w) < 0.2) . However, when used for the desiccation of L . bulgaricus, these parameters achieved a viability of less than 10%.

Int J Syst Evol Microbiol, 2004 Sep, 54(Pt 5), 1903 - 4
The status of the species Lactobacillus rogosae Holdeman and Moore 1974 . Request for an opinion; Felis GE et al.; A brief history of the species Lactobacillus rogosae is presented . It was ascertained that the type strain and other existing strains are not available in any established culture collection; therefore, they cannot be included in any scientific study . This matter is referred to the Judicial Commission, asking for an opinion on the status of the species.

Int J Syst Evol Microbiol, 2004 Sep, 54(Pt 5), 1621 - 6
Lactobacillus curvatus subsp . melibiosus is a later synonym of Lactobacillus sakei subsp . carnosus; Koort J et al.; On the basis of phenotypic and DNA-DNA reassociation studies, strain CCUG 34545T has been considered to represent a distinct Lactobacillus curvatus subspecies, Lactobacillus curvatus subsp . melibiosus . However, in several independent studies dealing with Lactobacillus sakei and L . curvatus strains, the subspecies division of L . curvatus has been found to be controversial . The original study distinguishing the two subspecies within both L . curvatus and L . sakei also lacked 16S rRNA gene sequence analyses . Therefore, the taxonomic position of L . curvatus subsp . melibiosus CCUG 34545T was re-evaluated in a polyphasic taxonomy study that included 16S rRNA gene sequence analysis, DNA-DNA reassociation, DNA G + C content determination, numerical analysis of ribotypes and whole-cell protein patterns and the examination of some fundamental phenotypic properties . The results obtained indicate that strain CCUG 34545T and its duplicate, CCUG 41580T, are Lactobacillus sakei subsp . carnosus strains and that L . curvatus subsp . melibiosus is a later synonym of L . sakei subsp . carnosus.

Dig Dis Sci, 2004 Aug, 49(7-8), 1095 - 102
Probiotics reduce bacterial colonization and gastric inflammation in H . pylori-infected mice; Johnson-Henry KC et al.; Probiotics are characterized by their ability to interact with commensal microflora in the gastrointestinal tract to produce beneficial health effects . In vitro studies suggest that Lactobacillus species have the potential to suppress the growth of Helicobacter pylori . The goal of this study was to determine if pretreatment of mice with a commercial mixture of live probiotics (L . rhamnosus, strain R0011, and L . acidophilus, strain R0052) would suppress colonization of H . pylori, strain SS1 . Thirty C57BL/6 female mice were divided into four groups: Group A was fed sterile water, group B received probiotics in sterile drinking water, group C was challenged orogastrically with H . pylori, and group D was pretreated with probiotics in drinking water prior to and following challenge with H . pylori . Rectal swabs, stomach homogenates, and luminal contents from ileum and colon were plated onto colistin nalidixic acid plates . Serial dilutions of stomach homogenates were plated onto H . pylori-sensitive agar plates and incubated under microaerophilic conditions . Tissue samples from the stomach were analyzed histologically to determine the degree of H . pylori colonization, mucosal inflammation, and epithelial cell apoptosis . Probiotics in drinking water did not affect the overall well-being of mice . Lactobacillus species were excreted in stools over the entire duration of treatment . Pretreatment with probiotics reduced the number of mice with H . pylori growth from stomach homgenates (100 to 50%; P = 0.02) . The percentage of mice with moderate-severe H . pylori-induced inflammation in the gastric antrum was reduced with probiotic pretreatment (71 to 29%; P = 0.14) . However, pretreatment with probiotics did not prevent H . pylori-induced apoptosis in the gastric mucosa . This preparation of probiotics provided a safe and novel approach for reducing H . pylori colonization and bacterial-induced inflammation of mice.

Infect Immun, 2004 Oct, 72(10), 5750 - 8
NF-kappaB- and AP-1-mediated induction of human beta defensin-2 in intestinal epithelial cells by Escherichia coli Nissle 1917: a novel effect of a probiotic bacterium; Wehkamp J et al.; Little is known about the defensive mechanisms induced in epithelial cells by pathogenic versus probiotic bacteria . The aim of our study was to compare probiotic bacterial strains such as Escherichia coli Nissle 1917 with nonprobiotic, pathogenic and nonpathogenic bacteria with respect to innate defense mechanisms in the intestinal mucosal cell . Here we report that E . coli strain Nissle 1917 and a variety of other probiotic bacteria, including lactobacilli--in contrast to more than 40 different E . coli strains tested--strongly induce the expression of the antimicrobial peptide human beta-defensin-2 (hBD-2) in Caco-2 intestinal epithelial cells in a time- and dose-dependent manner . Induction of hBD-2 through E . coli Nissle 1917 was further confirmed by activation of the hBD-2 promoter and detection of the hBD-2 peptide in the culture supernatants of E . coli Nissle 1917-treated Caco-2 cells . Luciferase gene reporter analyses and site-directed mutagenesis experiments demonstrated that functional binding sites for NF-kappaB and AP-1 in the hBD-2 promoter are required for induction of hBD-2 through E . coli Nissle 1917 . Treatment with the NF-kappaB inhibitor Helenalin, as well as with SP600125, a selective inhibitor of c-Jun N-terminal kinase, blocked hBD-2 induction by E . coli Nissle 1917 in Caco-2 cells . SB 202190, a specific p38 mitogen-activated protein kinase inhibitor, and PD 98059, a selective inhibitor of extracellular signal-regulated kinase 1/2, were ineffective . This report demonstrates that probiotic bacteria may stimulate the intestinal innate defense through the upregulation of inducible antimicrobial peptides such as hBD-2 . The induction of hBD-2 may contribute to an enhanced mucosal barrier to the luminal bacteria.

J Mol Microbiol Biotechnol, 2004, 7(4), 170 - 81
Glycerol metabolism of Lactobacillus rhamnosus ATCC 7469: cloning and expression of two glycerol kinase genes; Alvarez Mde F et al.; Lactobacillus rhamnosus ATCC 7469 was able to grow in glycerol as the sole source of energy in aerobic conditions, producing lactate, acetate, and diacetyl . A biphasic growth was observed in the presence of glucose . In this condition, glycerol consumption began after glucose was exhausted from the culture medium . Glycerol kinase activity was detected in L . rhamnosus ATCC 7469, a characteristic of microorganisms which catabolize glycerol in aerobic conditions . Genetic analysis revealed that this strain possesses two glycerol kinase genes: gykA and glpK, that encode for two different glycerol kinases GykA and GlpK, respectively . The glpK geneis associated in an operon with alpha-glycerophosphate oxidase (glpO) and glycerol facilitator (glpF) genes . Transcriptional analysis revealed that only glpK is expressed when L . rhamnosus was grown on glycerol .

Can J Microbiol, 2004 Jul, 50(7), 482 - 8
Lactobacillus casei, Lactobacillus rhamnosus, and Lactobacillus zeae isolates identified by sequence signature and immunoblot phenotype; Dobson CM et al.; Species taxonomy within the Lactobacillus casei group of bacteria has been unsettled . With the goal of helping clarify the taxonomy of these bacteria, we investigated the first 3 variable regions of the 16S rRNA gene, the 16S-23S rRNA interspacer region, and one third of the chaperonin 60 gene for Lactobacillus isolates originally designated as L . casei, L . paracasei, L . rhamnosus, and L . zeae . For each genetic region, a phylogenetic tree was created and signature sequence analysis was done . As well, phenotypic analysis of the various strains was performed by immunoblotting . Both sequence signature analysis and immunoblotting gave immediate identification of L . casei, L . rhamnosus, and L . zeae isolates . These results corroborate and extend previous findings concerning these lactobacilli; therefore, we strongly endorse recent proposals for revised nomenclature . Specifically, isolate ATCC 393 is appropriately rejected as the L . casei type strain because of grouping with isolates identified as L . zeae . As well, because all other L . casei isolates, including the proposed neotype isolate ATCC 334, grouped together with isolates designated L . paracasei, we support the use of the single species L . casei and rejection of the name L . paracasei.

J Reprod Immunol, 2004 Oct, 63(2), 111 - 22
Inoculation of Lactobacillus expressing hCG beta in the vagina induces an anti-hCG beta antibody response in murine vaginal mucosa; Yao XY et al.; OBJECTIVES: To test the possibility of vaccination with lactobacillus expressing hCG beta antigen administered by vaginal mucosal immunization . METHODS: A plasmid pIlac-hCG beta was constructed and then transfected into Lactobacillus casei CECT5276, which stably expressed hCG beta protein . RIA was used to detect hCG beta in the culture supernatant and cell lysate . Western blotting was performed to evaluate the expressed protein of interest . Female BALB/c mice aged 6-8 weeks received inoculations in the vagina of the recombinant L . casei CECT5276 . ELISA was used to determine the anti-hCG beta IgA antibody in vaginal lavage fluid from the BALB/c mice after vaginal mucosal immunization . RESULTS: The pIlac alone appeared to have a higher efficiency than pIlac-hCG beta, and the highest transfection efficiency of both plasmids was at pulse voltages of 1200 V and 1500 V . About 78.5% of the hCG beta protein was excreted into the culture supernatant . Excretion of hCG beta was most efficient when the pH of the culture medium was adjusted to around 7.0 and the concentration of lactose was around 1% . The hCG beta protein in the vaginal lavage fluid of these BALB/c mice was positive on the third day after vaginal inoculation . Anti-hCG beta IgA antibody continued to be found in the vaginal lavage fluid for 2 weeks following a booster vaginal inoculation . The splenic lymphocytes of the mice immunized with hCG beta through the vagina underwent a proliferative reaction to hCG antigen restimulation in vitro . Interferon gamma (IFN-gamma) and interleukin (IL)-4 were secreted at higher levels after vaginal mucosal immunization of L . casei expressing hCG beta than after vaginal mucosal immunization of L . casei alone . CONCLUSIONS: Vaginal immunization of lactobacillus expressing hCG beta induced an anti-hCG beta antibody response in the murine vaginal mucosa . Induction of the antigen-specific antibodies in the reproductive tract following vaginal inoculation of recombinant lactobacillus will lead to the development of a safe, efficient, and easy-to-use form of immunocontraception.

Clin Nutr, 2004 Oct, 23(5), 1069 - 73
An investigation into the effect of a probiotic on gut immune function in surgical patients; Woodcock NP et al.; BACKGROUND: The gut-associated lymphoid tissue (GALT) is an important component of the gut barrier . We have previously demonstrated a significant increase in various parameters of gut immune function in association with bacterial translocation . Animal studies have suggested that the probiotic Lactobacillus plantarum 299v improves the immunological status of the intestinal mucosa . The aim of this study was to determine whether the same is true in humans . METHOD: This was a prospective randomised controlled study, in which immunohistochemical techniques were used to measure the concentrations of plasma cells, IgA positive cells and IgM positive cells in the lamina propria, together with the concentrations of IgA and IgM at the mucosal surface in specimens of normal small bowel obtained from patients undergoing elective abdominal surgery who had consumed an oral preparation containing the probiotic Lactobacillus plantarum 299v (ProViva) during the immediate preoperative period . These were compared with similar specimens obtained from control subjects who did not receive the probiotic . RESULTS: A total of 22 patients were studied (probiotic group n = 11, control group n = 11) . The median volume of ProViva consumed was 3250 ml (range 2100-9000 ml), for a median duration of 9 days (range 5-18 days) . There were no significant differences between the probiotic and control groups in terms of concentrations of plasma cells, IgA positive cells or IgM positive cells in the lamina propria . There was a significantly higher concentration of IgM at the mucosal surface in the control group (P = 0.02, Fishers Exact test mid P), but no difference in terms of IgA . CONCLUSIONS: The increase in IgA observed in the intestinal mucosa in response to probiotics in animal studies does not occur in humans . The significance of the increase in IgM at the mucosal surface in the controls is unclear.

Res Microbiol, 2004 Oct, 155(8), 655 - 61
Bacterial desorption in water-saturated porous media in the presence of rhamnolipid biosurfactant; Chen G et al.; We investigated the effects of transients in elution chemistry on bacterial desorption in water-saturated porous media . Two typical Gram-positive bacterial strains of Lactobacillus casei and Streptococcus mitis were used as the model bacteria in this research . These two strains were first deposited in the porous medium, after which the medium with deposited bacteria was flushed with rhamnolipid biosurfactant solutions with a step increase in concentrations, and pulse-type bacterial releases were obtained . Bacterial desorption was quantified from bacterial breakthrough curves . It was found that bacterial retention in silica sand corresponded to bacterial interaction free energies with silica sand evaluated at the equilibrium distance, which were calculated based on independently determined bacterial, sediment and solution surface thermodynamic properties . With the increase in rhamnolipid biosurfactant concentrations, interactions between bacteria and silica sand decreased, and consequently less bacteria were retained . The decrease in interactions between bacteria and silica sand with increasing rhamnolipid biosurfactant concentrations was attributed to a decrease in the solution electron acceptor parameter of the Lewis acid/base component of surface tension, {Formula: see text} . The increase in rhamnolipid biosurfactant concentrations favored the decrease in solution {Formula: see text}, and consequently decreased the interactions between bacteria and silica sand.

Arch Microbiol, 2004 Nov, 182(5), 337 - 45 Epub 2004 Sep 14.
Random transposon mutagenesis of Lactobacillus plantarum by using the pGh9:IS S1 vector to clone genes involved in the regulation of phenolic acid metabolism; Gury J et al.; The lactic acid bacterium Lactobacillus plantarum possesses a promising inducible padA promoter that controls the expression of the padA gene encoding a phenolic acid decarboxylase, and which is transcriptionally regulated by phenolic acids . A strategy was followed in order to clone genes involved in the transcriptional regulation of the padA gene . The pGh9:IS S1 plasmid was used to perfect the mutagenesis of L . plantarum by transposition . This plasmid transposed randomly in the L . plantarum NC8 chromosome, with a frequency of 0.03% at a non-permissive replicating temperature of 42 degrees C . A L . plantarum mutant strain harbouring the transcriptional fusion padA:cat that displayed a chloramphenicol acetyl transferase (CAT) activity induced by p-coumaric acid was constructed . This strain was mutated by transposition with pGh9:IS S1 in order to select mutants with a constitutive CAT activity therefore able to grow on medium containing chloramphenicol without the phenolic acid inducer . Four mutants were identified and three of them harboured an inserted single copy of the pGh9:IS S1 vector . Analysis of IS S1 target sites allowed the identification of two genes: one encoding a putative protein that displays similarity with histidine-binding protein, the other belongs to the ATP-dependent RNA helicase family, protein which are usually involved in gene expression.

J Bacteriol, 2004 Oct, 186(19), 6661 - 6
Major role of NAD-dependent lactate dehydrogenases in aerobic lactate utilization in Lactobacillus plantarum during early stationary phase; Goffin P et al.; NAD-independent lactate dehydrogenases are commonly thought to be responsible for lactate utilization during the stationary phase of aerobic growth in Lactobacillus plantarum . To substantiate this view, we constructed single and double knockout mutants for the corresponding genes, loxD and loxL . Lactate-to-acetate conversion was not impaired in these strains, while it was completely blocked in mutants deficient in NAD-dependent lactate dehydrogenase activities, encoded by the ldhD and ldhL genes . We conclude that NAD-dependent but not NAD-independent lactate dehydrogenases are involved in this process.

J Bacteriol, 2004 Oct, 186(19), 6647 - 50
Dynamic cell surface hydrophobicity of Lactobacillus strains with and without surface layer proteins; Vadillo-Rodriguez V et al.; Variations in surface hydrophobicity of six Lactobacillus strains with and without an S-layer upon changes in ionic strength are derived from contact angle measurements with low- and high-ionic-strength aqueous solutions . Cell surface hydrophobicity changed in response to changes in ionic strength in three out of the six strains, offering these strains a versatile mechanism to adhere to different surfaces . The dynamic behavior of the cell surface hydrophobicity could be confirmed for two selected strains by measuring the interaction force between hydrophobic and hydrophilic tips with use of atomic force microscopy.

J Agric Food Chem, 2004 Sep 8, 52(18), 5735 - 9
Antioxidant activity of fermented soybean extract; Hu CC et al.; Free radicals are considered to be important causative factors in the development of cancer and cardiovascular diseases . This relationship has led to interest in evaluating the antioxidant capacities of many dietary supplements . Fermented soybean extract is produced by symbiotic fermentation of organic soybean with 20 types of Lactobacillus and yeast . In vitro and in vivo models are used in this study to evaluate the antioxidant effect of fermented soybean extract . Several in vitro models are used to detect the antioxidant capacity of the fermented soybean extract, which is compared to vitamin C and Trolox . The results demonstrate that the fermented soybean extract has strong antioxidant activity against unsaturated fatty acid peroxidation compared to vitamin C and Trolox . By the means of the test system developed by Y . Toshiki et al., it is shown that the fermented soybean extract can function both as an antioxidant and as a free radical acceptor that can convert free radicals into harmless substances through an energy-decreasing procedure . An in vivo study examines the effects of fermented soybean extract on the activity of antioxidant enzymes . The activities of the antioxidant enzymes (AOE) including total superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) are measured in liver, kidney, and brain from male Sprague-Dawley rats . The activities of CAT, SOD, and GPX are increased in the liver . However, the SOD activity is decreased in the kidney . SOD and GPX activities are decreased in the brain . These results lead to the conclusion that fermented soybean extract not only has antioxidant activity but also has an effect on the activity of antioxidant enzymes in liver.

Food Addit Contam, 2004 Jul, 21(7), 626 - 31
Fumaric acid in apple juice: a potential indicator of microbial spoilage of apples used as raw material; Gokmen V et al.; The effects of heat treatment (evaporation and excessive heating), and microbial contamination (Rhizopus stolonifer, Penicillium expansum and Lactobacillus plantarum) as sources of fumaric acid formation in apple juice concentrates were investigated . Fumaric acid formed in apple juice did not exceed 1.0 mg l(-1) during both commercial-scale evaporation and laboratory-scale excessive heat treatment, indicating that malic dehydration is not the primary source of fumaric acid formation . However, R . stolonifer and L . plantarum produced 18.23 +/- 0.82 and 5.39 +/- 0.31 mg l(-1) fumaric acid as determined in the raw juice obtained from contaminated apples after 5 days of incubation . The mean fumaric acid content of apple juice concentrates manufactured in 2000, 2001 and 2002 were determined as 4.9 +/- 1.9, 5.7 +/- 2.8 and 4.1 +/- 2.6 mg l(-1), respectively . The overall results suggest that the primary source of fumaric acid in apple juice concentrate is the use of apples decayed by certain microorganisms capable of producing fumaric acid.

Acta Odontol Scand, 2004 Jun, 62(3), 143 - 6
Composition of the salivary microflora during habitual consumption of fluoridated milk; Engstrom K et al.; The aim was to evaluate the effect of habitual consumption of fluoridated milk on the composition of the salivary microflora . The study group comprised 20 healthy schoolchildren and young adults with a mean age of 13.6 years and the investigation had a randomized double-blind crossover design with a washout period of 1 month . After professional tooth-cleaning at baseline, the subjects were supplied with either fluoridated (250 mL, 5 ppm F) or non-fluoridated milk for one daily intake during a period of 4 weeks . Salivary samples were collected immediately before tooth-cleaning and after 1, 2, and 4 weeks, respectively . The samples were immediately cultivated for total viable counts, oral streptococci, mutans streptococci, lactobacilli, and actinomyces spp . Bacterial counts were logarithmically transformed before statistical evaluation using ANOVA . No significant alterations of the salivary microflora were found during any of the milk regimens compared with baseline . There was a slight reduction in the proportion of mutans streptococci after 2 and 4 weeks during consumption with fluoridated milk but the difference failed to reach statistical significance . In conclusion, this study was unable to disclose any significant alteration of the composition of the salivary microflora following daily intake of fluoridated milk.

J Microbiol Methods, 2004 Nov, 59(2), 271 - 81
Identification by fluorescence spectroscopy of lactic acid bacteria isolated from a small-scale facility producing traditional dry sausages; Ammor S et al.; Three different fluorescence spectra were recorded following excitation at 250 nm (aromatic amino acids+nucleic acids, AAA+NA), 316 nm (NADH) and 380 nm (FAD) for 20 type strain collections of lactic acid bacteria (LAB) . Evaluation of the data using principal component analysis and factorial discriminant analysis showed a good discrimination of considered LAB at the genus, species and genus-species level . AAA+NA fluorophores showed the highest percentage of good classification . From AAA+NA spectra recorded on LAB isolated from a small-scale facility producing traditional dry sausages, we succeeded to identify 28 of 29 wild strains . This method allowed us to discriminate between Lactobacillus sakei subsp . carnosus and Lactobacillus sakei subsp . sakei . Thus, intrinsic fluorescence is an economical and powerful tool for the identification of wild LAB isolated from meat and meat products.

J Microbiol Methods, 2004 Nov, 59(2), 149 - 62
FT-IR spectroscopy for identification of closely related lactobacilli; Oust A et al.; Fourier Transform Infrared (FT-IR) spectroscopy was used to analyse 56 strains from four closely related species of Lactobacillus, L . sakei, L . plantarum, L . curvatus and L . paracasei . Hierarchical Cluster Analysis (HCA) was used to study the clusters in the data, but in the dendrogram, the spectra were not differentiated into four separate clusters corresponding to species . When the data were analysed with Partial Least Squares Regression (PLSR), the strains were differentiated into four clusters according to species . It was also possible to recognise strains that were incorrectly identified by conventional methods prior to the FT-IR analysis . PLSR was used to identify strains from three of the species, and the results were compared to two other multivariate methods, Soft Independent Modelling of Class Analogy (SIMCA) and K-Nearest Neighbour (KNN) . The three methods gave equally good identification results . The results show that FT-IR spectroscopy in combination with PLSR, or other multivariate methods, is well suited for identification of Lactobacillus at the species level, even in quite large data sets.

Gerodontology, 2004 Sep, 21(3), 130 - 40
Ten-year cross-sectional and incidence study of coronal and root caries and some related factors in elderly Swedish individuals; Fure S; OBJECTIVE: The 10-year incidence of dental caries was related to some associated factors in a random sample of 65, 75 and 85-year-old inhabitants of Gothenburg . SUBJECTS: Of the 208 persons examined at baseline, 102 (49%) participated in the follow-up examination; 56, 37 and nine, respectively, in the different age groups . For the purpose of time-trend comparisons, a new random sample of 98 individuals aged 55 years was examined . RESULTS: Ninety-five per cent of the participants had developed one or more carious lesions during the 10-year period and the incidence of coronal and root caries increased with age . In the 65-year-olds, 9% of the root surfaces had decayed during the period, compared with 25% in the 85-year-olds . Secondary caries predominated over primary caries and prosthetic crowns accounted for 70% of the restored tooth surfaces . Twenty per cent of the individuals were daily smokers and 61% were taking drugs with hyposalivatory side-effects . The mean saliva secretion rates were lower in the older groups compared with the 'younger' ones . The overall salivary counts of mutans streptococci and lactobacilli had increased during the period and the values were highest in the oldest age groups . Salivary levels of lactobacilli and mutans streptococci, number of teeth, daily numbers of cigarettes and drugs and oral hygiene were the best predictors of the incidence of caries . CONCLUSION: The findings indicate that there is an increased risk of dental caries with age owing to unfavourable caries-related factors.

Syst Appl Microbiol, 2004 Aug, 27(4), 443 - 53
Technological and molecular diversity of Lactobacillus plantarum strains isolated from naturally fermented sourdoughs; Pepe O et al.; Thirty Lactobacillus (L.) plantarum strains, isolated from sourdough, were identified by biochemical tests as well as 16S rDNA sequencing and differentiated on the basis of technological properties, such as amylase, protease, phytase and antirope activities . These properties were shown to be widely differing among the strains, indicating a significant technological diversity . Genetic differentiation was achieved by restriction endonuclease analysis-pulsed field gel electrophoresis (REA-PFGE) that allowed the L . plantarum strains to be divided into 10 different genomic groups . Moreover, 32 different starters were employed in dough making experiments; each starter consisted of a single strain of L . plantarum associated with a maltose positive or a maltose negative yeast . The technological properties of the doughs were greatly influenced by the type of strain included in the starter . The time of leavening and the acidification activities detected in the dough were enhanced by the presence of L . plantarum strains . The bacterial and yeast contents and fermentation properties were statistically treated by principal component analysis (PCA), which allowed the discrimination of different typologies of dough . The study of the peculiar characteristics of different strains of L . plantarum is fundamental for a better understanding of their potential in affecting the nutritional value, quality and stability of the baked goods . L . plantarum strains are able to differentially influence the dough quality when employed as starters.

J Clin Pediatr Dent, 2004 Summer, 28(4), 327 - 32
Mutans streptococci, lactobacilli in saliva and acidity from organisms in dental plaque: changes after restorative treatment; Morinushi T et al.; The effect of restorative dental treatment was evaluated on mutans streptococci and lactobacilli in saliva, as well as acid production by plaque bacteria . We used semiquantitative culture kits at time points up to 6 months following treatment . Changes in eating habits and oral hygiene during the study period were ruled out using a questionnaire . Mutans streptococci, lactobacilli, and acidity all had decreased significantly at 1 week after treatment . Acidity often was the first variable to return to pretreatment levels, while abundant at 6 than 3 months . Lactobacilli showed the most durable response to treatment . Restorative treatment achieves import temporary decreases in caries-associated bacteria, especially lactobacilli, without influence from potentially relevant behavioral changes . In addition, follow up examinations at relatively long intervals at least 3 month would appear most effective for dental health management.

Quintessence Int, 2004 Sep, 35(8), 649 - 52
Sports dentistry: buccal and salivary profile of a female soccer team; de Sant'Anna GR et al.; Sports dentistry has been considered a prominent area of dentistry because dental health can limit the abilities of athletes, both professional and nonprofessional, in their training and competition . Dental decay is associated with the frequent use of carbohydrates, recommended as an energy source for exercise . Strong indications exist regarding the possibility to use saliva as a performance determinant and for evaluation and prescription of physical activity . This study evaluated the salivary profiles (pH, flow rate, mutans streptococci and Lactobacillus counts) and decayed, missing, and filled teeth of 18 female soccer players (13 to 19 years old) of the Olympic Training and Research Center of Sao Paulo, before and after a training session . The salivary flow rate presented a significant reduction after training; however, there was no significant alteration in pH . Fifty percent of the players presented 10(5) to 10(6) mutans streptococci, and 66% presented 10(3) Lactobacillus . Several salivary components protect against microorganisms that cause superior respiratory tract infections (common in athletes), as well as participating in the remineralization mechanism during cariogenic challenges . Thus, due to the salivary flow rate reduction in this population with a high number of cariogenic microorganisms, noncariogenic drink ingestion at regular intervals and maintenance of hydration levels during training, are suggested.

Int J Food Microbiol, 2004 Nov 1, 96(2), 199 - 204
Tannase activity by lactic acid bacteria isolated from grape must and wine; Vaquero I et al.; We examined a range of oenological lactic acid bacteria species and reference strains for their potential to degrade tannins . Bacterial tannase activity was checked by a spectrophotometric and a visual reading method . None of the strains belonging to the oenological species of the genus Lactobacillus, Leuconostoc, Oenococcus or Pediococcus were tannase producers, with the exception of Lactobacillus plantarum . All the L . plantarum strains analyzed were positive for tannase activity and their identities were reconfirmed by L . plantarum PCR-specific assay or by sequencing the 16S rDNA . Tannase activity could be considered an important criterion for the selection of malolactic starter cultures since it might confer advantages in the winemaking process by reducing astringency and haze in wine.

Int J Food Microbiol, 2004 Nov 1, 96(2), 149 - 64
Evaluation of meat born lactic acid bacteria as protective cultures for the biopreservation of cooked meat products; Vermeiren L et al.; In this study, 91 strains, originating from meat products, were subjected to a step-by-step screening and characterisation to search for potential protective cultures to be used in the cooked cured meat industry . Strains were first tested on their homofermentative and psychrotrophic character and salt tolerance . Secondly, the antibacterial capacities towards Listeria monocytogenes, Leuconostoc mesenteroides, Leuconostoc carnosum and Brochotrix thermosphacta were determined in an agar spot test . In total, 38% of the tested strains were inhibitory towards all indicator strains . However, 91%, 88% and 74% of the strains could inhibit, respectively, L . monocytogenes, B . thermosphacta and Leuc . mesenteroides . Finally, 12 strains, with the highest antibacterial capacities, were evaluated on their competitive nature by comparing their growth rate, acidifying character and lactic acid production at 7 degrees C under anaerobic conditions in a liquid broth . All 12 strains, except for a bacteriocin producing Lactobacillus plantarum strain and the lactocin S producing Lactobacillus sakei 148, combined a fast growth rate with a deep and rapid acidification caused by the production of high levels of lactic acid . The 12 selected strains were then further investigated for their growth capacity on a model cooked ham product to establish whether the presence of these cultures on the ham did not negatively influence the sensory properties of the ham . All strains grew in 6 days at 7 degrees C from a level of 10(5)-10(6) to 10(7)-10(8) cfu/g and again the bacteriocin producing L . plantarum strain was the slowest growing strain . As the glucose level of the model cooked ham product was low (0.09+/-0.03%), growth of the putative protective cultures resulted in glucose depletion and a limited lactic acid production and accompanying pH decrease . Cooked ham inoculated with isolates 13E, 10A, 14A (all three identified as L . sakei subsp . carnosus by SDS-PAGE) and with strains L . sakei 148 (LS5) and L . sakei subsp . carnosus SAGA 777 (LS8) were not rejected by the sensory panel at the 34th day of the vacuum packaged storage at 7 degrees C . Therefore, these strains could have potential for the use as protective culture in cooked meat products.

FEMS Immunol Med Microbiol, 2004 Oct 1, 42(2), 173 - 80
Patterns of cytokine induction by gram-positive and gram-negative probiotic bacteria; Cross ML et al.; Bacteria used in commercial probiotic preparations are most commonly gram-positive lactic acid-producing species, although there are also some probiotic products which utilise gram-negative coliform bacteria . Characterising how the innate immune system responds to these bacteria in vitro may give an indication as to the likely immunomodulatory events that can be triggered following probiotic administration in vivo . Here, an established gram-positive probiotic (Lactobacillus casei Shirota) was compared against a novel gram-negative probiotic strain (Escherichia coli Nissle 1917) for its ability to induce cytokine production in a cell type representative of the innate immune system; in addition, responses were contrasted against those induced by an enteropathogenic coliform, E . coli 2282 . We investigated the ability of these three bacterial strains to modulate production of interleukins-10, -12 and -18; tumour necrosis factor-alpha; interferon-alpha; and transforming growth factor-beta, via a series of in vitro culture experiments involving the murine monocyte/macrophage cell line J774A.1 . All bacteria induced marked secretion of IL-12 and TNFalpha by cells, while only coliforms induced production of IL-10; there was minimal or no induction of IL-18 or TGFbeta . Activation of cells with recombinant gamma-interferon promoted increased production of IL-12, but decreased production of IL-10, in response to the co-culture of coliform bacteria, indicating differential cytokine induction depending on the activation status of the target cell . In general, live bacteria stimulated higher levels of IL-10, IL-12 and TNFalpha secretion than heat-killed preparations, while only live coliforms induced IFNalpha . These findings are discussed in relation to the likely immunomodulatory effects of gram-positive and gram-negative bacteria on the innate immune system in vivo, with particular emphasis on the marked similarity in cytokine response patterns observed between probiotic versus pathogenic coliform bacteria.

Bioresour Technol, 2005 Jan, 96(1), 63 - 7
Joint effect of nitrogen sources and B vitamin supplementation of date juice on lactic acid production by Lactobacillus casei subsp . rhamnosus; Nancib A et al.; The use of date juice as a substrate for lactic acid production was investigated . Various nitrogen sources were compared with yeast extract for efficient lactic acid production by Lactobacillus casei subsp . rhamnosus . Among different nitrogen sources added to date juice (yeast extract, ammonium sulfate, tryptic soy, urea, peptone, and casein hydrolysate), yeast extract was the most efficient . The effect of yeast extract could have been due to its B vitamin content . The addition of five B vitamins at less than 25 mg/l to date juice with any nitrogen source enhanced lactic acid production to some extent, except for date juice with yeast extract or urea or peptone . The most significant increase was obtained with ammonium sulfate . Half of the yeast extract content (10 g/l) in a supplemented date juice could be replaced by a mixture of B vitamins at less than 25 mg/l, and ammonium sulfate at 2.6 g/l with no significant decrease in lactic acid production.

J Pharmacol Sci, 2004 Sep, 96(1), 84 - 90 Epub 2004 Sep 10.
Yogurt containing Lactobacillus gasseri OLL2716 exerts gastroprotective action against acute gastric lesion and antral ulcer in rats; Uchida M et al.; Yogurt containing Lactobacillus gasseri OLL2716 (LG21 yogurt) is reported to improve Helicobacter pylori-induced gastric mucosal inflammation in clinical studies . However, other beneficial effects of LG21 yogurt have not been clarified . Therefore, we examined whether LG21 yogurt exhibits a gastroprotective action against acute gastric lesion or antral ulcer in rats . Moreover, the mechanism of gastroprotective action was also evaluated . After fasting, acute gastric lesions were induced by 0.6 M HCl . Gastric mucosal folds were stained by oral administration of methyl violet . Antral ulcers were induced by the combined administration of diethyldithiocarbamate and HCl in refed rats after fasting . LG21 yogurt was orally administered before HCl treatment or staining the mucosal folds . LG21 yogurt significantly and dose-dependently inhibited the formation of acute gastric lesions, and this gastroprotective action was attenuated by pretreatment with indomethacin . LG21 yogurt also significantly increased prostaglandin E2 generation in the gastric mucosa . Stained length of gastric mucosal fold was reduced by LG21 yogurt . Antral ulcer formation was also significantly inhibited by LG21 yogurt . From the above results, it was found that the ingestion of LG21 yogurt is useful for the prevention of gastric ulcer . Moreover, endogenous prostaglandin was suggested to be one of the gastroprotective mechanisms of LG21 yogurt.

Int J Food Microbiol, 2004 Oct 1, 96(1), 97 - 102
The microflora of fermented nixtamalized corn; Sefa-Dedeh S et al.; Nixtamalization is a traditional process that improves the nutritional quality of corn . To provide a means of utilizing the nutritional benefits of nixtamalized corn and improve product acceptability, lactic acid fermentation was applied . The objective of the study was to study the microbial profile and establish the important lactobacilli of fermenting nixtamalized corn dough . Two batches of cleaned whole corn were subjected to the process of nixtamalization, using two concentrations of lime (0.5 or 1.0%), milled, made into a dough (50% moisture) and fermented spontaneously for 72 h . A control sample was prepared without alkaline treatment . pH and titratable acidity of the dough were measured . Aerobic mesophiles, lactic acid bacteria, yeasts and molds were enumerated on Plate Count Agar (PCA), deMan, Rogossa and Sharpe (MRS) Agar and Malt Extract Agar (MEA), respectively . The identity of lactobacilli present was established at the species level using API 50 CHL . The pH of all the fermenting systems decreased with fermentation time with concomitant increase in titratable acidity . Lactic acid bacteria in numbers of 1.6 x 10(9), 2.3 x 10(9) and 1.8 x 10(9) cfu/g, respectively yeasts and molds, and numbers of 8.0 x 10(7), 5.0 x 10(5) and 1.7 x 10(5) cfu/g, respectively were observed in the control and the two nixtamalized (0.5% and 1.0% lime) samples after 48 h of fermentation . Lactobacilli identified in the fermenting nixtamalized corn dough were Lactobacillus plantarum, Lactobacillus fermentum and Lactobacillus cellobiosus as well as Pediococcus spp . The study demonstrates that nixtamalized corn though alkaline in nature can be subjected to spontaneous fermentation to produce a sour product .

Int J Food Microbiol, 2004 Oct 1, 96(1), 61 - 6
Transition of the probiotic bacteria, Lactobacillus casei strain Shirota, in the gastrointestinal tract of a pig; Ohashi Y et al.; The transition of probiotic bacteria Lactobacillus in the gastrointestinal tract was investigated in pigs that received commercially available fermented milk prepared with Lactobacillus casei strain Shirota (LCS) . Three female pigs fistulated at the cecum were fed 130 ml of fermented milk that contained over 10(10) (cfu) LCS with their daily meal for 8 days . Cecal contents were sampled through a fistula every 2 h for 24 h after marker dosing . The viable cell number (log cfu/g) of LCS and the concentrations of transit markers in each sample were determined . The viable number of LCS cells ranged from 3.56 to 6.58 . The number of LCS in the cecum was not stable in pigs and varied with the flow of the cecal content . The viable number of LCS cells was significantly correlated with the relative concentration of the marker . These results indicated that most LCS moved with the liquid component . The level of LCS reached the maximum (6.38) 6 h after dosing . Four doses every 6 h may be required to maintain the maximum level of LCS at the cecum .

FEMS Microbiol Lett, 2004 Sep 15, 238(2), 291 - 5
Low pH-induced membrane fatty acid alterations in oral bacteria; Fozo EM et al.; Four oral bacterial strains, of which two are considered aciduric and two are considered acid-sensitive, were grown under glucose-limiting conditions in chemostats to determine whether their membrane fatty acid profiles were altered in response to environmental acidification . Streptococcus gordonii DL1, as well as the aciduric strains S . salivarius 57.I, and Lactobacillus casei 4646 increased the levels of mono-unsaturated membrane fatty acids . The non-aciduric strain S . sanguis 10904 did not alter its membrane composition in response to pH values examined here . Thus, in response to low pH, aciduric oral bacteria alter their membrane composition to contain increased levels of long-chained, mono-unsaturated fatty acids . This suggests that membrane fatty acid adaptation is a common mechanism utilized by bacteria to withstand environmental stress.

J Appl Microbiol, 2004, 97(4), 876 - 81
Reduction of alpha-galactooligosaccharides in soyamilk by Lactobacillus fermentum CRL 722: in vitro and in vivo evaluation of fermented soyamilk; LeBlanc JG et al.; AIMS: Consumption of soya-derived products has been hampered by the presence of alpha-galactooligosaccharides (alpha-GOS) because mammals lack pancreatic alpha-galactosidase (alpha-Gal) which is necessary for their hydrolysis . These sugars thus reach the large intestine causing gastrointestinal disorders in sensitive individuals . The use of lactic acid bacteria (LAB) expressing alpha-Gal is a promising solution for the degradation of alpha-GOS in soyamilk . METHODS AND RESULTS: The capacity of the LAB Lactobacillus fermentum CRL 722 to properly degrade alpha-GOS was studied in vitro using controlled fermentation conditions and in vivo using a rat model . Lactobacillus fermentum CRL 722 was able to grow on commercial soyamilk and completely eliminated stachyose and raffinose during fermentation because of its high alpha-Gal activity . Rats fed soyamilk fermented by this LAB had smaller caecums compared with rats fed unfermented soyamilk . CONCLUSIONS: Soyamilk fermentation by Lact . fermentum CRL 722 results in the reduction of alpha-GOS concentrations in soyamilk, thus eliminating possible undesirable physiological effects normally associated with its consumption . SIGNIFICANCE AND IMPACT OF THE STUDY: Fermentation with Lact . fermentum CRL 722 could prevent gastrointestinal disorders in sensitive individuals normally associated with the consumption of soya-based products . This LAB could thus be used in the elaboration of novel fermented vegetable products which better suit the digestive capacities of consumers.

J Appl Microbiol, 2004, 97(4), 818 - 26
The effect of Propionibacterium acidipropionici, with or without Lactobacillus plantarum, on the fermentation and aerobic stability of wheat, sorghum and maize silages; Filya I et al.; AIMS: To determine the effect of Propionibacterium acidipropionici, alone or in combination with Lactobacillus plantarum, on the fermentation and aerobic stability of wheat, sorghum and maize silages . METHODS AND RESULTS: The inoculants were applied at 1.0 x 10(6) CFU g(-1) . Silages with no additives served as control . Fresh forages were sampled prior to ensiling . Three jars per treatment were sampled on days 2, 4, 8, 16 and 60 after ensiling, for chemical and microbiological analysis . At the end of the ensiling period, the silages were subjected to an aerobic stability test . The P . acidipropionici-inoculated silages had significantly higher levels of acetic and propionic acid than the L . plantarum or P . acidipropionici + L . plantarum-inoculated silages (P < 0.05) . Therefore, yeast activity was impaired in the P . acidipropionici-inoculated silages . As a result, P . acidipropionici decreased CO(2) production and improved aerobic stability of wheat, sorghum and maize silages . However, the combination of P . acidipropionici + L . plantarum did not improve aerobic stability of the silages . CONCLUSIONS: The P . acidipropionici was very effective in protecting the wheat, sorghum and maize silages exposed to air under laboratory conditions, probably because the acidic environment under ensiling conditions is favourable for this micro-organism . SIGNIFICANCE AND IMPACT OF THE STUDY: The use of P . acidipropionici, as a silage inoculant can improve the aerobic stability of silages by inhibition of yeast activity.

J Appl Microbiol, 2004, 97(4), 774 - 82
Cloning, molecular characterization and expression analysis of two small heat shock genes isolated from wine Lactobacillus plantarum; Spano G et al.; AIM: Understanding the molecular response to stress tolerance of wine Lactobacillus plantarum . METHODS AND RESULTS: Two genes codifying for heat shock proteins were cloned from wine L . plantarum . The coding regions of the two heat shock genes are 420 and 444 nucleotides long, and started with an ATG codon suggesting that they were translated . The protein sequences deduced from the isolated genes have a molecular mass of 18.483 and 19.282 kDa, respectively, and were therefore named hsp18.5 and hsp19.3 . The expression of small heat shock genes was analysed by RT-PCR analysis . Moreover, the 5' and 3' noncoding regions were cloned and sequenced . CONCLUSIONS: The expression of the heat shock genes was strongly induced by heat, cold and ethanol stress . Analysis of the 5' and 3' flanking regions of hsp18.5 and hsp19.3 genes, revealed the presence of an inverted repeat sequence (TTAGCACTC-N(9)-GAGTGCTAA) homologue to the CIRCE elements found to the upstream regulatory region of heat shock operons, and an inverted sequence that could form a stem and loop structure that it is likely to function as a transcriptional terminator . Based on their structures, the genes were classified as belonging to Class I of heat shock genes according to the B . subtilis nomenclature of heat response genes . SIGNIFICANCE AND IMPACT OF THE STUDY: Small heat shock genes isolated from wine L . plantarum might have a role in preventing damage by cold stress.

J Appl Microbiol, 2004, 97(4), 749 - 56
The aggregation-promoting factor of Lactobacillus crispatus M247 and its genetic locus; Marcotte H et al.; AIMS: Characterization of the aggregation-promoting factor (APF) of the human intestinal isolate Lactobacillus crispatus M247 and its homologous nonaggregating mutant Mu5 . METHODS AND RESULTS: Western blot analysis revealed that the supernatant of both M247 and Mu5 contains a 28-kDa protein which cross reacts with the antiserum produced against the APF of Lact . gasseri 4B2 . The apf genes of M247 and Mu5 strains were identical and were shown to be 672 nucleotides in length and encoding a protein of 223 amino acids with a predicted molecular weight of 24.0 kDa . CONCLUSION: Our results shows that the lost of aggregation in Mu5 is not related to a defect in secretion of the APF protein or a mutation in the apf gene . SIGNIFICANCE AND IMPACT OF THE STUDY: These results suggest that the mutation in Mu5 may be contained in another molecule involved in aggregation such as a possible receptor for APF.

J Appl Microbiol, 2004, 97(4), 712 - 8
Genetic marker for differentiating beer-spoilage ability of Lactobacillus paracollinoides strains; Suzuki K et al.; AIMS: To determine whether the beer-spoilage ability is an intrinsic character of Lactobacillus paracollinoides and identify a genetic marker for differentiating the beer-spoilage ability of strains belonging to this species . METHODS AND RESULTS: The ribotype of a nonspoilage strain, Lact . brevis ATCC8291, was found to be identical with that of Lact . paracollinoides LA7 . The 16S rDNA sequence analysis and DNA-DNA hybridization study indicates that nonspoilage ATCC8291 should belong to Lact . paracollinoides . We further isolated nonspoilage variants from Lact . paracollinoides LA2(T) and LA9 by incubating these strains at 30 degrees C . To identify a genetic marker for differentiating the beer-spoilage ability of Lact . paracollinoides, open reading frames 5 (ORF5), the previously reported genetic marker for Lact . brevis, was evaluated . As a result, ORF5 homologues were detected in all of the 12 beer-spoilage strains of Lact . paracollinoides, while this ORF was not found in ATCC8291 or the two nonspoilage variants obtained from LA2(T) and LA9 . CONCLUSIONS: Lactobacillus paracollinoides is not an intrinsic beer-spoiler and the nonspoilage strain Lact . brevis ATCC8291 should be reclassified as Lact . paracollinoides . ORF5 was found to be useful for differentiating beer-spoilage ability of this species . SIGNIFICANCE AND IMPACT OF THE STUDY: The finding that Lact . paracollinoides includes nonspoilage strains necessitates brewers to use a genetic marker that is associated with the beer-spoilage ability of this species.

J Appl Microbiol, 2004, 97(4), 673 - 81
Role of viability of probiotic strains in their persistence in the gut and in mucosal immune stimulation; Galdeano CM et al.; AIMS: To determine how probiotic bacteria contact with intestinal epithelial and immune cells and the conditions to induce a good mucosal immune stimulation . METHODS AND RESULTS: Lactobacillus casei was studied by transmission electron microscopy (TEM) to determine its interaction with the gut . We compared the influence of viable and nonviable lactic acid bacteria on the intestinal mucosal immune system (IMIS) and their persistence in the gut of mice . TEM showed whole Lact . casei adhered to the villi; the bacterial antigen was found in the cytoplasm of the enterocytes . Viable bacteria stimulated the IMIS to a greater extent than nonviable bacteria with the exception of Lact . delbrueckii subsp . bulgaricus . For all the strains assayed at 72 h no antigenic particles were found in the intestine . CONCLUSION: Antigenic particles but not the whole bacteria can enter to epithelial cells and contact with the immune cells . Bacterial viability is a condition for a better stimulation of the IMIS . SIGNIFICANCE AND IMPACT OF THE STUDY: We demonstrated that only antigenic particle interact with the immune cells and their fast clearance from the gut agrees with those described for the particulate antigens . The regular consumption of probiotics should not adversely affect the host.

J Microbiol, 2004 Jun, 42(2), 133 - 8
Free fatty acid accumulation by mesophilic lactic acid bacteria in cold-stored milk; Coskun H et al.; This study was aimed to determine the accumulation of free fatty acid by mesophilic lactic acid bacteria (Lactococcus lactis subsp . lactis 1471, Lactococcus lactis subsp . cremoris 1000 and Lactobacillus casei 111) in cold-stored milk . According to the results, all cold-stored milks had higher acid degree values than those of fresh milk . This phenomenon showed that a slight increase occurred in the accumulation of free fatty acids as a result of spontaneous lipolysis during cold storage . All lactic acid bacteria showed good performance in production of titratable acidity, which increased during fermentation of the milk (fresh and stored milks) . Moreover, as the storage time was prolonged, more free fatty acid accumulation was obtained from the fermentation of the cold-stored milk by the investigated lactic acid bacteria . The control milk, which was without lactic acid bacteria, showed no change in the accumulation of free fatty acid during fermentation . From this result, it can be suggested that longer cold-storage time can induce higher free fatty acid accumulation in milk by lactic acid bacteria.

Lett Appl Microbiol, 2004, 39(4), 336 - 40
Malt sprout extract medium for cultivation of Lactobacillus plantarum protective cultures; Laitila A et al.; AIMS: The aim was to develop a cheap cereal-based alternative medium for the large-scale production of biopreservative Lactobacillus plantarum VTT E-79098 . We examined the effect of growth medium and pH control on the cell yield of Lact . plantarum E-79098 and the antimicrobial activity of the cell-free extracts . METHODS: Fermentations using a novel Malt Sprout Extract Medium (MSE) were performed with different pH regimes . The antimicrobial activity of the cell-free extracts against Pantoea agglomerans VTT E-90396 and Fusarium avenaceum VTT D-80147 was assessed with automated turbidometry . SIGNIFICANCE AND IMPACT OF THE STUDY: When compared with MRS, the MSE medium cultures produced equal growth yields of Lact . plantarum VTT E-79098 and enhanced antimicrobial potential against the Gram-negative bacterium P . agglomerans and a Fusarium fungus . The MSE medium can be used as a low-cost alternative to MRS for producing high cell yields and good antimicrobial activity of Lact . plantarum.

Lett Appl Microbiol, 2004, 39(4), 313 - 8
Purification and characterization of an endopeptidase that has an important role in the carboxyl terminal processing of antihypertensive peptides in Lactobacillus helveticus CM4; Ueno K et al.; AIMS: To purify and characterize a peptidase that can catalyse C-terminal processing of antihypertensive peptide from Lactobacillus helveticus CM4 . METHODS AND RESULTS: An endopeptidase which seems to process the carboxyl terminal end of two antihypertensive peptides, Val-Pro-Pro and Ile-Pro-Pro, was purified from Lactobacillus helveticus CM4 by four stages of column chromatography, using synthetic pro-peptide as a substrate . The molecular weight of the purified enzyme was estimated to be 67,000 by SEPHACRYL S-200 and 70,000 by SDS-PAGE analysis . The purified enzyme generated: (i) Val-Pro-Pro from Val-Pro-Pro-Phe-Leu and Val-Pro-Pro-Phe-Leu-Gln-Pro, and (ii) Ile-Pro-Pro from Ile-Pro-Pro-Leu-Thr and Ile-Pro-Pro-Leu-Thr-Gln-Thr, but theses peptides could not be generated from Val-Pro-Pro-Phe, Val-Pro-Pro-Phe-Leu-Gln, Ile-Pro-Pro-Leu and Ile-Pro-Pro-Leu-Thr-Gln . Part of the amino terminal sequence of the purified enzyme had homology to a previously reported pepO gene product . CONCLUSION: These results suggest that the purified endopeptidase isolated in this study have an important role in the carboxyl terminal processing of two antihypertensive peptides in Lact . helveticus CM4.

J Mater Sci Mater Med, 2002 Jan, 13(1), 125 - 131
Study of a hydraulic calcium phosphate cement for dental applications; Serraj S et al.; Calcium phosphate-based cements (CPCs) have attracted much interest because of their good osteoconductivity for bone reconstruction . We obtained CPCs by mixing calcium bis-dihydrogenophosphate monohydrate (MCPM) and calcium oxide with water or sodium phosphate buffers (NaP) as liquid phase . Cement samples with different calcium-to-phosphate ratios (Ca/P), liquid-to-powder ratios (L/P) and liquid phases were analyzed by X-rays diffraction (XRD), pH-metry, extensometry and calorimetry . Antibacterial activity on two bacterial strains (Streptococcus mutans, Lactobacillus acidophilus) and a polycontaminated bacterial inoculum was also studied using the agar diffusion method . The best mechanical properties ( approximately 25 MPa) corresponded to Ca/P ratios between 1.67 and 2.5, a 1 M sodium phosphate buffer pH 7, as liquid phase and a L/P ratio of 0.6\,\hbox{ml g}^{\,-\,1} . The final setting time increased with the Ca/P ratio . The setting expansion, around 1-2%, depended on the Ca/P and L/P ratios . The inner temperature of the cements rose to 45 degrees during setting then decreased rapidly . The injectability was 100% up to 3.5 min and then decreased . It increased with increasing the L/P ratio but to the detriment of the compressive strength and setting time . XRD analysis indicated that the setting reaction led to a mixture of calcium hydroxide and calcium-deficient hydroxyapatite even for a Ca/P ratio of 1.67 . Consequently, the pH of the surrounding fluids rose to 11.5-12 during their dissolution . Bacterial growth inhibition was only clearly observed for Ca/P>/=2 . This bioactive calcium phosphate cement can potentially be employed for pulp capping and cavity lining as classical calcium hydroxide-based cements, but it is not usable, in the present formulation, for root canal filling because of its short setting time.

Appl Environ Microbiol, 2004 Sep, 70(9), 5315 - 22
Identification and inactivation of genetic loci involved with Lactobacillus acidophilus acid tolerance; Azcarate-Peril MA et al.; Amino acid decarboxylation-antiporter reactions are one of the most important systems for maintaining intracellular pH between physiological limits under acid stress . We analyzed the Lactobacillus acidophilus NCFM complete genome sequence and selected four open reading frames with similarities to genes involved with decarboxylation reactions involved in acid tolerance in several microorganisms . Putative genes encoding an ornithine decarboxylase, an amino acid permease, a glutamate gamma-aminobutyrate antiporter, and a transcriptional regulator were disrupted by insertional inactivation . The ability of L . acidophilus to survive low-pH conditions, such as those encountered in the stomach or fermented dairy foods, was investigated and compared to the abilities of early- and late-stationary-phase cells of the mutants by challenging them with a variety of acidic conditions . All of the integrants were more sensitive to low pH than the parental strain . Interestingly, each integrant also exhibited an adaptive acid response during logarithmic growth, indicating that multiple mechanisms are present and orchestrated in L . acidophilus in response to acid challenge.

Appl Environ Microbiol, 2004 Sep, 70(9), 5081 - 8
Influence of complex nutrient source on growth of and curvacin a production by sausage isolate Lactobacillus curvatus LTH 1174; Verluyten J et al.; Lactobacillus curvatus LTH 1174, a fermented sausage isolate, produces the antilisterial bacteriocin curvacin A . Its biokinetics of cell growth and bacteriocin production as a function of various concentrations of a complex nutrient source were investigated in vitro during laboratory fermentations with modified MRS medium . A modification of the nutrient depletion model was used to fit the data describing growth and bacteriocin production . Both cell growth and bacteriocin activity were influenced by changes in the complex nutrient source concentration . Standard MRS medium clearly limited the growth of L . curvatus LTH 1174 . Higher nutrient concentrations, up to a certain degree, led to improved growth, a higher attainable biomass concentration, and a higher bacteriocin activity in the supernatant . A lower concentration of complex nutrient source caused severe growth inhibition, leading to a lower biomass concentration but a much higher specific bacteriocin production . When examining the separate components of the complex nutrient source, a stimulating effect of bacteriological peptone on growth was found without an adverse effect on bacteriocin production, resulting in increased curvacin A activity . Furthermore, specific depletion of the amino acids tyrosine, serine, and asparagine/aspartic acid was observed for this strain.

BMC Pediatr . 2004 Sep 02;4(1):18.
Lactobacillus casei strain GG in the treatment of infants with acute watery diarrhea: a randomized, double-blind, placebo controlled clinical trial {ISRCTN67363048}; Salazar-Lindo E et al.; BACKGROUND: Adjuvant therapy to ORT with probiotic bacteria for infants with acute watery diarrhea has been under active investigation . Most studies have been done in the developed world showing benefit only for viral mild gastroenteritis . We evaluated the effect of a milk formula containing one billion (109) cfu/ml of Lactobacillus casei strain GG (LGG) upon duration and severity of diarrhea in infants in an environment with more severe acute diarrhea, where etiologic agents other than rotavirus are involved more frequently, and where mixed infections are more prevalent . METHODS: Male infants aged 3-36 months brought for treatment of acute watery diarrhea of less than 48 hours were eligible . After rehydration was completed with the WHO's oral rehydration solution, patients were randomly assigned to receive a milk formula either containing LGG or not . Stool volume was periodically measured using a devise suited to collect stools separate from urine . Duration of diarrhea was estimated based on stools physical characteristics . RESULTS: Eighty nine patients received the placebo milk formula and ninety received the LGG containing formula . Both groups were comparable in their baseline characteristics . Total stool output was significantly larger (p = 0.047) in the LGG group (247.8 ml/kg) than in the placebo group (195.0 ml/kg) . No significant differences were found in duration of diarrhea (58.5 hours with LGG vs . 50.4 hours with placebo), rate of treatment failure (21.1% with LGG vs . 18.0% with placebo), and proportion of patients with unresolved diarrhea after 120 hours (12.2% with LGG vs . 12.5% with placebo) . The rate of stools with reducing substances after 24 hours of treatment increased significantly in both groups (from 41.4% to 72.2% with LGG and from 45.9% to 68.0% with placebo) . CONCLUSION: This study did not show a positive effect of LGG on the clinical course of acute watery diarrhea . Positive beneficial effects of LGG, as had been reported elsewhere, could have been masked in our study by worsening diarrhea due to transient lactose malabsorption . Further studies with low-lactose or non-lactose conveyors of LGG are desirable.

J Bacteriol, 2004 Sep, 186(18), 6059 - 69
Two arginine repressors regulate arginine biosynthesis in Lactobacillus plantarum; Nicoloff H et al.; The repression of the carAB operon encoding carbamoyl phosphate synthase leads to Lactobacillus plantarum FB331 growth inhibition in the presence of arginine . This phenotype was used in a positive screening to select spontaneous mutants deregulated in the arginine biosynthesis pathway . Fourteen mutants were genetically characterized for constitutive arginine production . Mutations were located either in one of the arginine repressor genes (argR1 or argR2) present in L . plantarum or in a putative ARG operator in the intergenic region of the bipolar carAB-argCJBDF operons involved in arginine biosynthesis . Although the presence of two ArgR regulators is commonly found in gram-positive bacteria, only single arginine repressors have so far been well studied in Escherichia coli or Bacillus subtilis . In L . plantarum, arginine repression was abolished when ArgR1 or ArgR2 was mutated in the DNA binding domain, or in the oligomerization domain or when an A123D mutation occurred in ArgR1 . A123, equivalent to the conserved residue A124 in E . coli ArgR involved in arginine binding, was different in the wild-type ArgR2 . Thus, corepressor binding sites may be different in ArgR1 and ArgR2, which have only 35% identical residues . Other mutants harbored wild-type argR genes, and 20 mutants have lost their ability to grow in normal air without carbon dioxide enrichment; this revealed a link between arginine biosynthesis and a still-unknown CO2-dependent metabolic pathway . In many gram-positive bacteria, the expression and interaction of different ArgR-like proteins may imply a complex regulatory network in response to environmental stimuli.

Obstet Gynecol, 2004 Sep, 104(3), 555 - 63
A comparison between the vaginal ring and oral contraceptives; Veres S et al.; OBJECTIVE: To investigate whether contraceptive vaginal ring use results in similar estimated genital symptoms, signs, examination, and laboratory findings compared with oral contraceptive use . METHODS: Women were randomly assigned to either contraceptive vaginal ring or a 20 microg ethinyl estradiol oral contraceptive pill use for 3 consecutive 28-day cycles, directly followed by 3 cycles of the study drug not initially assigned . Subjects scored genital symptoms on a daily diary using a 0-4 scale and underwent a baseline, cycle 2, cycle 4, and exit pelvic examination including vaginal discharge evaluation, vaginal Gram stain and white cell count, and culture for yeast and Lactobacillus, including colony count and hydrogen peroxide production . RESULTS: Of the 40 subjects assigned to each arm, 33 (82.5%) subjects in the ring-first arm and 31 (77.5%) subjects in the pill-first arm completed all study visits (P =.58) . Most subjects reported few genital symptoms with either method, but 63% of subjects reported vaginal wetness during ring use compared with 43% during pill use . During ring use larger numbers of Lactobacillus colonies present were positive for hydrogen peroxide production (fold difference 2.67, 95% confidence interval 1.49, 4.78, P <.001) . All other laboratory data, including yeast colony counts, Nugent Gram stain score, vaginal white blood cell count, vaginal pH, and discharge weight, were not significantly different by method . CONCLUSION: Some women may notice an increase in vaginal wetness during contraceptive ring use yet the method is well tolerated and appears to improve the vaginal flora.

J Ind Microbiol Biotechnol, 2004 Oct, 31(9), 401 - 8 Epub 2004 Oct.
Bacterial contaminants of fuel ethanol production; Skinner KA et al.; Bacterial contamination is an ongoing problem for commercial fuel ethanol production facilities . Both chronic and acute infections are of concern, due to the fact that bacteria compete with the ethanol-producing yeast for sugar substrates and micronutrients . Lactic acid levels often rise during bouts of contamination, suggesting that the most common contaminants are lactic acid bacteria . However, quantitative surveys of commercial corn-based fuel ethanol facilities are lacking . For this study, samples were collected from one wet mill and two dry grind fuel ethanol facilities over a 9 month period at strategic time points and locations along the production lines, and bacterial contaminants were isolated and identified . Contamination in the wet mill facility consistently reached 10(6) bacteria/ml . Titers from dry grind facilities were more variable but often reached 10(8)/ml . Antibiotics were not used in the wet mill operation . One dry grind facility added antibiotic to the yeast propagation tank only, while the second facility dosed the fermentation with antibiotic every 4 h . Neither dosing procedure appeared to reliably reduce overall contamination, although the second facility showed less diversity among contaminants . Lactobacillus species were the most abundant isolates from all three plants, averaging 51, 38, and 77% of total isolates from the wet mill and the first and second dry grind facilities, respectively . Although populations varied over time, individual facilities tended to exhibit characteristic bacterial profiles, suggesting the occurrence of persistent endemic infections.

Plasmid, 2004 Sep, 52(2), 84 - 8
Characterization of pC7 from Lactobacillus paraplantarum C7 derived from Kimchi and development of lactic acid bacteria--Escherichia coli shuttle vector; Park WJ et al.; A cryptic plasmid pC7 was characterized, which was isolated from Lactobacillus paraplantarum C7 derived from Kimchi, a traditional korean vegetable food . The plasmid pC7 is a circular molecule of 2,134 base-pairs in length with a G + C content of 38.5% . The nucleotide sequence analysis revealed the presence of an open reading frame encoding a putative 317 amino acids protein homologous to replication proteins RepA . Furthermore, a putative double and a single-strand origin were identified . Together with features of nucleotide sequences, the detection of single-stranded intermediate DNA in Lb . paraplantarum demonstrated that pC7 replicates via a rolling circle mechanism . A cloning vector for lactic acid bacteria was developed on the basis of the pC7 replicon, into which were inserted an erythromycin resistance gene as a marker, multiple cloning sites, and Escherichia coli ColE1 replication origin . E . coli and several species of Lactobacillus and Leuconostoc can be transformed with the resultant vector . Therefore, pC7 derivatives may be useful LAB-E . coli shuttle vectors, which are essential in engineering important strains in food fermentation.

FEMS Microbiol Lett, 2004 Sep 1, 238(1), 159 - 65
Genetic diversity (RAPD-PCR) of lactobacilli isolated from "Almagro" eggplant fermentations from two seasons; Sesena S et al.; Genetic diversity of 323 strains of lactobacilli isolated from an Almagro eggplant manufacturing plant was analyzed by using random amplified polymorphic DNA (RAPD) . Thirty-four distinct RAPD patterns were obtained with 95% of isolates grouped into 18 main clusters . Genetic diversity was higher in brines from season II, Lactobacillus plantarum and Lactobacillus fermentum/cellobiosus being the species with the greatest number of genotypes . A single L . fermentum/cellobiosus genotype comprised isolates from both seasons and could be considered endemic to that factory.

FEMS Microbiol Lett, 2004 Sep 1, 238(1), 9 - 15
Identification, characterisation and specificity of a cell wall lytic enzyme from Lactobacillus fermentum BR11; Turner MS et al.; Screening of a genomic library with an antiserum raised against whole Lactobacillus fermentum BR11 cells identified a clone expressing an immunoreactive 37-kDa protein . Analysis of the 3010-bp DNA insert contained within the clone revealed four open reading frames (ORFs) . One ORF encodes LysA, a 303 amino acid protein which has up to 35% identity with putative endolysins from prophages Lj928 and Lj965 from Lactobacillus johnsonii and Lp1 and Lp2 from Lactobacillus plantarum as well as with the endolysin of Lactobacillus gasseri bacteriophage Phiadh . The immunoreactive protein was shown to be encoded by a truncated ORF downstream of lysA which has similarity to glutamyl-tRNA synthetases . The N-terminus of LysA has sequence similarity with N-acetylmuramidase catalytic domains while the C-terminus has sequence similarity with putative cell envelope binding bacterial SH3b domains . C-terminal bacterial SH3b domains were identified in the majority of Lactobacillus bacteriophage endolysins . LysA was expressed in Escherichia coli and unusually was found to have a broad bacteriolytic activity range with activity against a number of different Lactobacillus species and against Lactococcus lactis, streptococci and Staphylococcus aureus . It was found that LysA is 2 and 8000 times more active against L . fermentum than L . lactis and Streptococcus pyogenes, respectively.

Ann Nutr Metab, 2004, 48(4), 228 - 34 Epub 2004 Aug 27.
Effects of long-term intervention with Lactobacillus helveticus-fermented milk on bone mineral density and bone mineral content in growing rats; Narva M et al.; BACKGROUND: Lactobacillus helveticus-fermented milk has been shown to increase calcium absorption compared to ordinary sour milk . In the present study the possible effect of L . helveticus-fermented milk on bone was studied in growing rats . METHODS: Spontaneously hypertensive male rats, which develop an osteoporotic bone disorder with age, were randomized into 5 groups (n = 10) receiving milk fermented with L . helveticus 16H and, as reference drinks, milk fermented with L . helveticus and Saccharomyces cerevisiae, sour milk, skim milk or water, for 14 weeks . After the intervention bone mineral density and bone mineral content were measured by dual-energy X-ray absorptiometry . The femur weight, length and volume were measured before ashing . From the ashes the weight and mineral content were assessed . RESULTS: As the body weight gain differed significantly between the groups, the results were related to the body weight . The L . helveticus-fermented milk intervention significantly increased the bone mineral density and bone mineral content compared to the sour milk, skim milk and water interventions . The mean values of the bone mineral density and bone mineral content were higher in the L . helveticus-fermented milk group compared to the Saccharomyces-fermented milk group but the difference was not statistically significant . CONCLUSION: L . helveticus-fermented milk increases bone mineral density and bone mineral content in relation to body weight in the long-term feeding of growing rats . The mechanism of L . helveticus remains to be discovered . Copyright (c) 2004 S . Karger AG, Basel.

BMJ . 2004 Sep 4;329(7465):548 . Epub 2004 Aug 27.
Effect of lactobacillus in preventing post-antibiotic vulvovaginal candidiasis: a randomised controlled trial; Pirotta M et al.; OBJECTIVE: To test whether oral or vaginal lactobacillus can prevent vulvovaginitis after antibiotic treatment . DESIGN: Randomised, placebo controlled, double blind, factorial 2x2 trial . SETTING: Fifty general practices and 16 pharmacies in Melbourne, Australia . PARTICIPANTS: Non-pregnant women aged 18-50 years who required a short course of oral antibiotics for a non-gynaecological infection: 278 were enrolled in the study, and results were available for 235 . INTERVENTIONS: Lactobacillus preparations taken orally or vaginally, or both, from enrollment until four days after completion of their antibiotic course . MAIN OUTCOME MEASURES: Participants' reports of symptoms of post-antibiotic vulvovaginitis, with microbiological evidence of candidiasis provided by a self obtained vaginal swab . RESULTS: Overall, 55/235 (23% (95% confidence interval 18% to 29%)) women developed post-antibiotic vulvovaginitis . Compared with placebo, the odds ratio for developing post-antibiotic vulvovaginitis with oral lactobacillus was 1.06 (95% confidence interval 0.58 to 1.94) and with vaginal lactobacillus 1.38 (0.75 to 2.54) . Compliance with antibiotics and interventions was high . The trial was terminated after the second interim analysis because of lack of effect of the interventions . Given the data at this time, the chances of detecting a significant reduction in vulvovaginitis with oral or vaginal lactobacillus treatment were less than 0.032 and 0.0006 respectively if the trial proceeded to full enrollment . CONCLUSIONS: The use of oral or vaginal forms of lactobacillus to prevent post-antibiotic vulvovaginitis is not supported by these results . Further research on this subject is unlikely to be fruitful, unless new understandings about the pathogenesis of post-antibiotic vulvovaginitis indicate a possible role for lactobacillus.

J Chromatogr A, 2004 Jul 23, 1043(2), 195 - 200
Agarose-coated anion exchanger prevents cell-adsorbent interactions; Viloria-Cols ME et al.; A common problem during recovery of bioproducts by adsorption from particulate broths is fouling of the adsorbent material as a result of the interaction of cells and cell debris, which present negative charges, with the positively charged anion exchangers commonly used in bioprocesses . The effect of shielding an adsorbent with a layer of agarose on reducing the binding of cells while still allowing the low-molecular-mass bioproducts to be adsorbed was studied . Coating the anion-exchange resin Amberlite IRA-400 with agarose followed by cross-linking the agarose layer effectively prevented the binding of Escherichia coli, Saccharomyces cerevisiae, and Lactobacillus casei cells but allowed binding of lactic acid to the adsorbent . The cross-linked agarose layer was stable during recycling of the adsorbent.

J Dairy Sci, 2004 Aug, 87(8), 2563 - 70
Microbial counts, fermentation products, and aerobic stability of whole crop corn and a total mixed ration ensiled with and without inoculation of Lactobacillus casei or Lactobacillus buchneri; Nishino N et al.; Whole crop corn (DM 29.2%) and a total mixed ration (TMR, DM 56.8%) containing wet brewers grains, alfalfa hay, dried beet pulp, cracked corn, soybean meal, and molasses at a ratio of 5:1:1:1:1:1 on fresh weight basis, were ensiled with and without Lactobacillus casei or Lactobacillus buchneri in laboratory silos . The effects of inoculation on microbial counts, fermentation products, and aerobic stability were determined after 10 and 60 d . Untreated corn silage was well preserved with high lactic acid content, whereas large numbers of remaining yeasts resulted in low stability on exposure to air . Inoculation with L . casei suppressed heterolactic fermentation, but no improvements were found in aerobic stability . The addition of L . buchneri markedly enhanced the aerobic stability, while not affecting the DM loss and NH3-N production . Large amounts of ethanol were found when the TMR was ensiled, and the content of ethanol overwhelmed that of lactic acid in untreated silage . This fermentation was related to high yeast populations and accounted for a large loss of DM found in the initial 10 d . The ethanol production decreased when inoculated with L . casei and L . buchneri, but the effects diminished at 60 d of ensiling . Inoculation with L . buchneri lowered the yeasts in TMR silage from the beginning of storage; however, the populations decreased to undetectable levels when stored for 60 d, regardless of inoculation . No heating was observed in TMR silage during aerobic deterioration test for 7 d . This stability was achieved even when a high population of yeasts remained and was not affected by either inoculation or ensiling period . The results indicate that inoculation with L . buchneri can inhibit yeast growth and improve aerobic stability of corn and TMR silage; however, high stability of TMR silage can be obtained even when no treatments were made and high population (>10(5) cfu/g) of yeasts were detected.

J Dairy Sci, 2004 Jul, 87(7), 2017 - 23
A new probiotic cheese with antioxidative and antimicrobial activity; Songisepp E et al.; The aim of our study was to develop an original probiotic cheese based on the Estonian open-texture, smear-ripened, semisoft cheese "Pikantne." Cheese was produced by two methods using cheese starter cultures (Probat 505) in combination with 0.04% of probiotic Lactobacillus fermentum strain ME-3 (10(9) cfu/mL) with high antimicrobial activity and antioxidative properties . The probiotic Lactobacillus was added into milk simultaneously with starter cultures (cheese A) and into drained curd (cheese B) . After addition of probiotic L . fermentum ME-3, the cheese composition, flavor, and aroma were comparable to the control cheese (score values = 4.5, 4.2, and 3.7 for control cheese, cheese A, and cheese B, respectively) . Cheese A, which had good sensory properties, was chosen for further testing of viability and probiotic properties . The probiotic strain was found to withstand the technological processing of cheese, surviving and sustaining moderate antimicrobial and high antioxidative activity throughout ripening and storage (the ripened cheese contained approximately 5 x 10(7) cfu/g viable ME-3 cells), although the viability of the ME-3 strain incorporated into the cheese showed a slight decrease between d 24 and 54 after cheese preparation . Semisoft cheese "Pikantne" serves as a suitable carrier of antimicrobial and antioxidative L . fermentum ME-3.

Oral Microbiol Immunol, 2004 Oct, 19(5), 281 - 8
Salivary IgA to cariogenic bacteria in HIV-positive children and its correlation with caries prevalence and levels of cariogenic microorganisms; Castro GF et al.; The interrelationship of HIV infection, dental caries and mucosal immune responses remains controversial . In our study population of 40 HIV-infected and 40 healthy control children (ages 2-5 years) there was a significantly higher prevalence of dental caries in HIV-infected children (P<0.05) . The extent of caries correlated with the severity of HIV disease . To determine whether the immunosuppression that ensues after HIV infection could contribute to the increased caries prevalence, the concentrations of total IgA and IgA specific to cariogenic bacteria (Streptococcus mutans, Streptococcus sobrinus and Lactobacillus acidophilus) were determined in whole saliva by enzyme-linked immunosorbent assay . Levels of the same bacteria were also quantified in saliva using checkerboard DNA-DNA hybridization . A significantly increased level of total salivary IgA was found in the HIV-positive population (P < 0.05), but there were comparable titers of specific IgA to cariogenic bacteria in HIV-positive and healthy controls . The microbiological assessment also demonstrated similar levels of cariogenic microorganisms in both groups . We conclude that HIV-positive children appear to maintain the capacity to mount a mucosal immune response to cariogenic microorganisms, at least until late stages of disease.

Huan Jing Ke Xue, 2004 May, 25(3), 69 - 72
{Flocculating mechanism of microbial flocculant MBFTRJ21}; Qin PY et al.; The bioflocculant MBFTRJ21 for nature soda was exploited in our lab, and its flocculating mechanism was studied . The bioflocculant produced by Lactobacillus, has some advantages, such as less dosage, better flocculating effects, and etc . MBFTRJ21 is mainly composed of polysaccharide, which is determined by Anthrone reaction, Coomassie blue reaction and Elson-Morgan method . The acting force between bioflocculant and nature soda is hydrogen bond, which is determined by xi electric potential, hydrogen bond and electrovalent bond determination . The bioflocculant is treated by heat and KIO4, which indicate the active ingredient of bioflocculant is protein and osamine . Bridge occur during flocculation process, which is determined by particle size distribution . The results showed that the flocculating mechanism of MBFTRJ21 resulted from the combination of nature soda with MBFTRJ21 by hydrogen bond at the active position of glycosaminoglycan.

Br Poult Sci, 2004 Jun, 45(3), 355 - 66
Immunomodulation by probiotic lactobacilli in layer- and meat-type chickens; Koenen ME et al.; 1 . The aim of the experiments was to evaluate whether selected probiotic lactobacillus strains have different immunomodulating effects in layer- and meat-type strain chickens . 2 . Humoral and cellular specific and non-specific immune responses were studied by experiments on cellular proliferation, entry and survival of Salmonella bacteria in gut and spleen leukocytes, immunoglobulin isotypes and specific immunoglobulin titres . 3 . The effects of two different feeding regimes (short and continuous feeding) and doses for administration of lactobacilli were studied . 4 . The lactobacillus strains that were evaluated showed modulating effects on the immune system of layer- and meat-type chickens . 5 . In meat-type strain chickens the lactobacilli had a stimulating effect when the chickens were young (up to 3 weeks) and the dose was relatively high, whereas in layer-type chickens a lower effective dose and discontinuous administration was also effective . 6 . Immunoprobiotic lactobacilli can have a positive effect on humoral and cellular immune responses in layer- and meat-type strain chickens, but the lactobacillus strain to be used, the age of the animals and effective dose of lactobacilli to be administered need to be optimised.

J Microbiol Methods, 2004 Oct, 59(1), 109 - 15
Optimising single cell activity assessment of Lactobacillus plantarum by fluorescent in situ hybridisation as affected by growth; de Vries MC et al.; Fluorescent in situ hybridisation (FISH) with a 16S ribosomal RNA (rRNA)-targeted oligonucleotide probe, Eub338, could be used to estimate the in situ activity of Lactobacillus plantarum WCFS1 in exponentially growing cells . However, L . plantarum is capable of growth to very high cell densities, and the properties of the L . plantarum cell envelope prevented effective entry of the fluorescent oligonucleotide probe into the cells at later stages of growth at high cell densities . Total rRNA measurements of cells isolated at different growth stages showed maximal amounts of RNA (8.77+/-0.8 fg) per cell at the early stationary phase and confirmed the effectiveness of FISH for accurate activity measurement in exponentially growing cells.

J Microbiol Methods, 2004 Oct, 59(1), 1 - 6
Identification of Lactobacillus sakei and Lactobacillus curvatus by multiplex PCR-based restriction enzyme analysis; Lee J et al.; Two closely related lactic acid bacteria, Lactobacillus sakei and Lactobacillus curvatus, are very difficult to be rapidly differentiated . Here we report multiplex polymerase chain reaction (PCR)-based restriction enzyme analysis that is useful for rapid and reliable identification of these two species . This method employs both polymerase chain reaction (PCR) and restriction enzyme analysis (REA) . First, multiplex-PCR using three primers that were designed from 16S rDNA sequence produces two bands, a 433-bp and a 623-bp band . A 433-bp band represents only L . sakei and L . curvatus among lactobacilli and genetically related bacteria, and a 623-bp band is used for further identification by restriction analysis . Second, restriction analysis of 623-bp band using Hind III restriction enzyme discriminates L . sakei from L . curvatus . This method could identify 28 strains as L . sakei or L . curvatus, which were frequently isolated from kimchi, a traditional fermented cabbage product in South Korea . Therefore, these results suggest that this method is simple, rapid, and reliable for the identification of L . sakei and L . curvatus species.

Biosci Biotechnol Biochem, 2004 Aug, 68(8), 1779 - 81
Stimulatory effects of casein phosphopeptide (CPP-III) on mRNA expression of cytokines in Caco-2 cells; Kawahara T et al.; The effect of CPP-III, a commercially available casein phosphopeptide, on the mRNA expression of cytokines in Caco-2 cells was investigated . CPP-III enhanced the mRNAs of interleukin (IL)-6 and tumor necrosis factor-alpha while IL-1beta was not affected . The mRNA expression of IL-6 was stronger in the presence of both CPP-III and bacterial components such as peptidoglycan from Lactobacillus acidophilus and lipopolysaccharide from Salmonella typhimurium . These results suggest that CPP-III influences the expression of cytokines in intestinal epithelial cells.

J Immunol, 2004 Sep 1, 173(5), 3492 - 503
Presence of IFN-gamma does not indicate its necessity for induction of coronary arteritis in an animal model of Kawasaki disease; Chan WC et al.; Kawasaki disease is the most common cause of vasculitis affecting children, and the leading cause of acquired heart disease in the developed world . To date, studies on the role of IFN-gamma in the pathogenesis of Kawasaki disease have focused on peripheral production of IFN-gamma, and have yielded conflicting results . Affected heart tissue is not available from children with Kawasaki disease . In this study, we use an animal model of Kawasaki disease, Lactobacillus casei cell wall extract (LCWE)-induced coronary arteritis, to examine the role of IFN-gamma in the development of coronary artery lesions . We report the presence of IFN-gamma, both at the mRNA and protein levels, in the affected vessels . Its biphasic expression, first at days 3-7 and again at days 28-42 post-LCWE injection, corresponds to the first appearance of inflammatory infiltrate in coronary arteries, and later to vascular wall disruption and aneurysm formation, respectively . Interestingly, ablation of IFN-gamma expression did not dampen the inflammatory response, and IFN-gamma-deficient lymphocytes proliferated more vigorously in response to LCWE than those of wild-type animals . Of more importance, the incidence of coronary arteritis was the same in IFN-gamma-deficient and wild-type mice . Taken together, our findings demonstrate that IFN-gamma regulates the immune response during development of coronary arteritis, but is not required for the induction of coronary artery disease.

Infect Immun, 2004 Sep, 72(9), 5308 - 14
Live Lactobacillus reuteri is essential for the inhibitory effect on tumor necrosis factor alpha-induced interleukin-8 expression; Ma D et al.; The mechanism of the apparent anti-inflammatory action of probiotic organisms is unclear . Lactobacillus reuteri is effective in inhibiting colitis in interleukin-10 (IL-10)-deficient mice . Nerve growth factor (NGF), in addition to its activity on neuronal cell growth, has significant anti-inflammatory effects in several experimental systems in vitro and in vivo, including a model of colitis . Our experiments were designed to explore the mechanism of effect of L . reuteri in the human epithelial cell lines T84 and HT29 on cytokine and NGF synthesis and IL-8 response to tumor necrosis factor alpha (TNF-alpha) . Epithelial cells were cultured for various times with live and killed L . reuteri and examined by reverse transcription-PCR for NGF, IL-10, and TNF-alpha-induced IL-8 expression . An enzyme-linked immunosorbent assay was used to quantitate intracellular IL-8 and secreted product . Western blotting and confocal microscopy were used to determine the effects on IkappaB and NF-kappaB, respectively . Live but not heat-killed or gamma-irradiated L . reuteri upregulated NGF and dose dependently inhibited constitutive synthesis by T84 and HT29 cells of IL-8 and that induced by TNF-alpha in terms of mRNA and intracellular and secreted protein . Similarly, L . reuteri inhibited IL-8 synthesis induced by Salmonella enterica serovar Typhimurium . L . reuteri required preincubation and adherence for effect, inhibited translocation of NF-kappaB to the nuclei of HeLa cells, and prevented degradation of IkappaB . Neither cellular lysates nor media supernatants had any effect on TNF-alpha-induced IL-8 . The conclusion is that L . reuteri has potent direct anti-inflammatory activity on human epithelial cells, which is likely to be related to the activity of ingested probiotics . L . reuteri also upregulates an unusual anti-inflammatory molecule, NGF, and inhibits NF-kappaB translocation to the nucleus.

FEMS Microbiol Lett, 2004 Aug 15, 237(2), 385 - 91
Nisin-controlled expression of Norwalk virus VP60 protein in Lactobacillus casei; Martin MC et al.; A food-grade strain with nisRK stably integrated into the genome, was constructed in order to implement the nisin-controlled expression system (NICE) in Lactobacillus casei ATCC393 . Expression of beta-glucuronidase (gus) reporter gene was employed to optimize the system, which has been successfully used to produce the main antigenic protein from Norwalk virus, opening new perspectives for producing edible vaccines.

Biomed Microdevices, 2004 Jun, 6(2), 125 - 30
Development of a novel hand-held immunoassay for the detection of enterohemorrhagic Escherichia coli O157:H7; Lin FY et al.; Escherichia coli O157:H7 is an important human pathogen responsible for numerous outbreaks of hemorrhagic colitis and hemolytic uremic syndrome world-wide . A portable detection device is needed for field testing and point-of-care testing . Poly(dimethylsiloxane) (PDMS) is a solid substrate well suited for adsorption of macromolecules . The purpose of this study was to develop a hand-held enzyme-linked immunosorbent assay (ELISA) for the detection of E . coli O157:H7 antigens . The prototype consisted of three modules: one for loading reagents, a second for immunosensor detection, and a third for discharge of wastes . Reagent delivery was achieved by using 1-ml syringes embedded within the loading module . The detection module was based on a PDMS layer on which varying concentrations of E . coli O157:H7 antigens, and negative controls (Lactobacillus rhamnosus R011 and phosphate-buffered saline) were passively adsorbed . Commercially available goat anti-E . coli O157:H7 antibody was used as a primary antibody, a donkey anti-goat IgG conjugated with horseradish peroxidase was used as a secondary antibody, and a precipitating substrate was employed for colorimetric detection . Results obtained with the prototype were compared to those obtained using a conventional nitrocellulose membrane-based immunoassay . Using the prototype, E . coli O157:H7 antigens, in quantities from 4 to 400 ng, were accurately detected . This detection limit was comparable to that observed using conventional dot-blot assays . The PDMS layer could be re-used without loss of sensitivity . Colorimetric detection could be visualized readily without the need for sophisticated equipment . Furthermore, this device can be adapted to perform diagnostics for other microbial pathogens currently detected using immunoassay methodology.

J Bacteriol, 2004 Sep, 186(17), 5906 - 18
High-resolution structure of the histidine-containing phosphocarrier protein (HPr) from Staphylococcus aureus and characterization of its interaction with the bifunctional HPr kinase/phosphorylase; Maurer T et al.; A high-resolution structure of the histidine-containing phosphocarrier protein (HPr) from Staphylococcus aureus was obtained by heteronuclear multidimensional nuclear magnetic resonance (NMR) spectroscopy on the basis of 1,766 structural restraints . Twenty-three hydrogen bonds in HPr could be directly detected by polarization transfer from the amide nitrogen to the carbonyl carbon involved in the hydrogen bond . Differential line broadening was used to characterize the interaction of HPr with the HPr kinase/phosphorylase (HPrK/P) of Staphylococcus xylosus, which is responsible for phosphorylation-dephosphorylation of the hydroxyl group of the regulatory serine residue at position 46 . The dissociation constant Kd was determined to be 0.10 +/- 0.02 mM at 303 K from the NMR data, assuming independent binding . The data are consistent with a stoichiometry of 1 HPr molecule per HPrK/P monomer in solution . Using transversal relaxation optimized spectroscopy-heteronuclear single quantum correlation, we mapped the interaction site of the two proteins in the 330-kDa complex . As expected, it covers the region around Ser46 and the small helix b following this residue . In addition, HPrK/P also binds to the second phosphorylation site of HPr at position 15 . This interaction may be essential for the recognition of the phosphorylation state of His15 and the phosphorylation-dependent regulation of the kinase/phosphorylase activity . In accordance with this observation, the recently published X-ray structure of the HPr/HPrK core protein complex from Lactobacillus casei shows interactions with the two phosphorylation sites . However, the NMR data also suggest differences for the full-length protein from S . xylosus: there are no indications for an interaction with the residues preceding the regulatory Ser46 residue (Thr41 to Lys45) in the protein of S . xylosus . In contrast, it seems to interact with the C-terminal helix of HPr in solution, an interaction which is not observed for the complex of HPr with the core of HPrK/P of L . casei in crystals.

J Bacteriol, 2004 Sep, 186(17), 5721 - 9
Identification of Lactobacillus plantarum genes that are induced in the gastrointestinal tract of mice; Bron PA et al.; Lactobacillus plantarum is a flexible and versatile microorganism that inhabits a variety of environmental niches, including the human gastrointestinal (GI) tract . Moreover, this lactic acid bacterium can survive passage through the human or mouse stomach in an active form . To investigate the genetic background of this persistence, resolvase-based in vivo expression technology (R-IVET) was performed in L . plantarum WCFS1 by using the mouse GI tract as a model system . This approach identified 72 L . plantarum genes whose expression was induced during passage through the GI tract as compared to laboratory media . Nine of these genes encode sugar-related functions, including ribose, cellobiose, sucrose, and sorbitol transporter genes . Another nine genes encode functions involved in acquisition and synthesis of amino acids, nucleotides, cofactors, and vitamins, indicating their limited availability in the GI tract . Four genes involved in stress-related functions were identified, reflecting the harsh conditions that L . plantarum encounters in the GI tract . The four extracellular protein encoding genes identified could potentially be involved in interaction with host specific factors . The rest of the genes are part of several functionally unrelated pathways or encode (conserved) hypothetical proteins . Remarkably, a large number of the functions or pathways identified here have previously been identified in pathogens as being important in vivo during infection, strongly suggesting that survival rather than virulence is the explanation for the importance of these genes during host residence.

J Nutr Health Aging, 2004, 8(4), 208 - 11
Monocyte function in healthy middle-aged people receiving fermented milk containing Lactobacillus casei; Parra D et al.; Background: Lactic acid bacteria have been suggested as a dietary strategy to enhance immune system activity . Objective: The aim of the current work was to test the effects of a Lactobacillus casei fermented milk consumption on monocyte activity of middle-aged volunteers . Design: Forty-five healthy volunteers, 24 women and 21 men (aged: 51 - 58 years), were randomized in two groups to receive three cups per day of a fermented milk containing L . casei DN114001 (108 - 1010/g) (n = 23), or placebo (n = 22), during 8 weeks . White blood cell count and the oxidative burst capacity of monocytes and granulocytes were examined with a FACScalibur . Measurements were performed at baseline and after the nutritional intervention, at day fifty-six . Results: After the trial, no changes in immune cell proportions were detected in both groups, as well as in monocyte activity after the placebo consumption (p = 0.625) . However, volunteers included in the probiotic-treated group increased (p = 0.029) their oxidative burst capacity of monocytes, and this increment inversely and significantly correlated with the intensity registered at baseline (r = -0.653, p = 0.004) . Conclusions: Results showed that daily intake of fermented milk containing Lactobacillus casei was able to module the oxidative burst capacity of monocyte subset in healthy middle-aged people, particularly in subjects with lower initial levels . Thus, this nutritional strategy could be considered to maintain immune competence in ageing.

Caries Res, 2004 Sep-Oct, 38(5), 436 - 41
Suppression of caries-related microorganisms in dentine lesions after short-term chlorhexidine or antibiotic treatment; Wicht MJ et al.; This study investigated the efficiency of a chlorhexidine varnish and an antibiotic paste in suppressing the cultivable microflora of deep dentine cavities in a stepwise excavation procedure . Subsequent to enamel preparation and removal of the central biomass, infected dentine was sampled from the cavity floor . Ten cavities each were either covered with the 1% chlorhexidine- and 1% thymol-containing varnish Cervitec (CE), the demeclocycline hydrocortisone-containing ointment Ledermix (LE) or received no treatment as control (CO) . A compomer composite was used as intermediate restorative . Cavities were reassessed after 6 weeks and again dentine samples were microbiologically investigated for total viable counts, mutans streptococci and lactobacilli . After 6 weeks a significant reduction of the total viable counts was observed in the LE group (p = 0.011) compared to the control, whereas no differences were found in the CE group (p > 0.05) . Mutans streptococci were rarely recovered at baseline and after 6 weeks . Compared to the CO group counts of lactobacilli were significantly reduced in the CE and LE groups (p < 0.05) . Lactobacillus species were frequently recovered at baseline and after 6 weeks of observation . Lactobacillus rhamnosus was the predominant species in all samples investigated . Application of CE or LE resulted in reduced counts of lactobacilli after a period of 6 weeks . Although none of the materials completely eliminated the viable microorganisms, the use of LE was more effective than CE in reducing the total anaerobic microorganisms associated with carious dentine.

J Agric Food Chem, 2004 Aug 25, 52(17), 5533 - 8
Structural characterization and biological activities of an exopolysaccharide kefiran produced by Lactobacillus kefiranofaciens WT-2B(T); Maeda H et al.; Lactobacillus kefiranofaciens, isolated from kefir grains, produces an extracellular polysaccharide when cultured, not only in PYG10 medium but also in a liquid medium containing a rice hydrolysate that had been previously degraded by treatment with a glucoamylase . The maximum yield of the polysaccharide, using the rice hydrolysate as the medium, was 2.5 g/L after a 7-day culture period at pH 5.0 and 33 degrees C . Compositional analysis, methylation analysis, specific rotation, and (1)H and (13)C NMR spectroscopy revealed that the structures of polysaccharides obtained from these two different culture media are essentially identical . The polysaccharide is composed of a hexasaccharide repeating unit and, thus, is known as kefiran . The weight-average molecular weight and the z-average radius of gyration of a sample, purified from the rice hydrolysate medium, were determined to be 7.6 x 10(5) g/mol and 39.9 nm, respectively, by gel permeation chromatography equipped with a multiangle laser-light-scattering photometer . Changes in blood pressure and serum components were examined in SHRSP/Hos rats, using doses of 100 and 300 mg of kefiran/kg of rat . A suppression in the increase in blood pressure was observed in these rats after 30 days . This activity is discussed in terms of the concentration of serum components of the rat, with emphasis on lipid components such as cholesterols, triglycerides, and free fatty acids.

Taehan Kanho Hakhoe Chi, 2004 Feb, 34(1), 132 - 40
{The effect of oral health education on oral health in kindergarten children.}; Song BS; PURPOSE: This study was performed to investigate the effects of oral health education on kindergarten children . METHOD: The study was designed as a quasi-experimental, nonequivalent control group pre -post test design . Data was collected from April 1st to November 30th, 2001 . The total samples were sixty seven healthy kindergarten children; the experiment group consisted of thirty three and the control group was thirty four children . The experimental group received an oral health education program daily for 20 minutes for four weeks . RESULT: There was significant improvements on oral health behavior in the experimental group . Streptococcus mutans and lactobacilli of the salivary variables in the experimental group were significantly higher than the control group . The dmft was lower in the experimental group than the control group, but there was no significant difference between the two groups . However it was significantly lower in the experimental group than control group over time . CONCLUSION: Oral health education for kindergarten children showed an increase in the use of tooth paste and practicing correct tooth brush usage . Also, it decreased the rate of eating cariogenic food and had a positive effect on oral health through suppressing dental cavities.

J Anim Sci, 2004 Jul, 82(7), 1976 - 85
Relative ileal amino acid flows and microbial counts in intestinal effluents of Goettingen Minipigs and Saddleback pigs are not different; Hennig U et al.; We explored the suitability of Goettingen Minipigs as models to measure ileal AA digestibility and evaluate dietary proteins for conventional pigs . Further, a potential for secondary ileal microbial colonization 5 mo after establishing end-to-end ileorectal anastomosis was investigated . Goettingen Minipigs (BW 18 kg) and Saddleback pigs (BW 27 kg) fitted with end-to-end ileorectal anastomosis were fed six diets based on barley and oilseed meals and three diets based on wheat and milk powder differing in total and ileal digestible lysine . Apparent ileal digestibilities of CP (N x 6.25) and of 20 AA were determined . No differences (P = 0.062 to 0.982) were found in AA apparent ileal digestibilities between breeds . Therefore, Minipigs are a reasonable model to estimate apparent ileal digestibility of AA for evaluation of dietary proteins . However, the apparent ileal digestibility of CP (P = 0.048) was higher in Minipigs than in Saddleback pigs (barley and oilseed meals-based diets 70% vs . 66%; wheat and milk powder-based diets 80% vs . 77%), which is probably due to a smaller contribution of non-AA-nitrogen in the ileal effluent of Goettingen Minipigs . For lysine, the apparent ileal digestibilities (means of both breeds) ranged from 78 to 85% in wheat and milk powder-based, and 70 to 78% in barley and oilseed-based diets . Experimentally derived concentrations of ileally digestible lysine confirmed the values predicted from a published table . Microbial counts were not affected by breed as shown for lactobacilli, with 9.1+/-0.2 and 9.1+/-0.2 (P = 0.977), enterococci with 4.8+/-0.3 and 5.6+/-0.4 (P = 0.162), and yeasts with 4.6+/-0.3 and 4.6+/-0.4 (P = 0.906) log cfu/g effluent for Goettingen Minipigs and Saddleback pigs, respectively . The counts did not change over 5 mo, suggesting that no secondary microbial colonization occurred in pigs with end-to-end ileorectal anastomosis.

Histochem Cell Biol, 2004 Aug, 122(2), 151 - 9
Selective expression of detoxifying glutathione transferases in mouse colon: effect of experimental colitis and the presence of bacteria; Edalat M et al.; Glutathione transferases (GSTs) play a central role in the cellular defense against harmful endogenous compounds and xenobiotics in mouse and man . The gastrointestinal channel is constantly exposed to bacteria, bacterial products, and xenobiotics . In the present study the distribution of alpha, mu, and pi class GSTs was examined immunohistologically in the colon of conventional and germ-free (GF) mice subjected to experimental colitis . The tissues samples were from conventional mice with and without colitis induced by dextran sulfate sodium (DSS); GF mice treated with DSS or carrageenan; and GF mice inoculated with normal mouse bacterial flora as well as with Lactobacillus GG . In conventional as well as in GF mice the mu and pi class GSTs showed reduced intestinal expression when colitis was induced . In con-rast, the level of GSTs reacting with antibodies directed against the alpha class, in particular mGST A4-4, was elevated after induction of inflammation . Of special interest is mGST A4-4 because of its high catalytic activity with toxic products of lipid peroxidation . In the colon of conventionalized GF mice that were given mouse intestinal flora, the mGST A4-4 expression was increased with time for several weeks, but then showed a decrease to a normal level . Additionally, the inoculation of GF mice with Lactobacillus GG induced all the intestinal GSTs studied.

Eur J Nutr, 2004 Dec, 43(6), 381 - 9 Epub 2004 Jul 14.
The effect of milk fermented by yogurt cultures plus Lactobacillus casei DN-114001 on the immune response of subjects under academic examination stress; Marcos A et al.; BACKGROUND : A suppressed immune response has been documented in students under examination stress . AIMS : The current study aimed to evaluate the effect of milk fermented with yogurt cultures plus Lactobacillus casei DN-114001 (Actimel((R))) on the immune system of subjects under academic examination stress . METHODS : University students were allocated to one of two groups, receiving during 6 weeks (3 weeks prior to, as well as the 3-week duration of the examination period) either: a) a glass of semi-skimmed milk each day (control group, n=63) or b) two 100mL portions per day of fermented milk (treatment group, n=73) . Anxiety and immunological measurements were monitored at baseline (Phase 0) and study end (Phase 1) . RESULTS : The results were expressed as the differences between the data obtained from Phase 0 and Phase 1 . This was calculated by subtracting Phase 1 results from the Phase 0 and it is denominated "Treatment effect" . Mean (+/- SE) anxiety increased significantly (P<0.05) over the 6-week study in all students, from 40.74+/-2.50 to 61.19+/-2.64 (in percentiles) . There was no significant treatment effect since this increase was similar in the control and the treatment groups (21.65+/-5.09 vs 19.14+/-3.67, respectively) . However, there was a significant treatment effect (P<0.05) on the mean change in absolute number of lymphocytes during the 6-week study, which decreased in the control group (-0.04+/-0.12 cells x 103/mm(3)) and increased in the treatment group (0.37+/-0.11 cells x 103/mm(3)) . There was also a significant treatment effect (P<0.05) on the change in absolute numbers of CD56 cells during the 6-week study . Mean absolute CD56 cells significantly decreased (P<0.05) in the control group (-51.97+/-21.33 cells/mm(3)),while remaining similar in the treatment group (17.29+/-17.27 cells/mm(3)) . During the study, mean serum cortisol increased 4.30+/-0.98 microg/dL in the control group, and 1.75+/-1.05 microg/dL in the treatment group and no significant differences were found between both values (P=0.062) . CONCLUSIONS : Milk fermented with yogurt cultures plus Lactobacillus casei DN-114001 was able to modulate the number of lymphocytes and CD56 cells in subjects under academic examination stress.

Anal Bioanal Chem, 2004 Sep, 380(1), 15 - 23 Epub 2004 Aug 07.
A rapid biosensor for viable B . anthracis spores; Baeumner AJ et al.; A simple membrane-strip-based biosensor assay has been combined with a nucleic acid sequence-based amplification (NASBA) reaction for rapid (4 h) detection of a small number (ten) of viable B . anthracis spores . The biosensor is based on identification of a unique mRNA sequence from one of the anthrax toxin genes, the protective antigen ( pag), encoded on the toxin plasmid, pXO1, and thus provides high specificity toward B . anthracis . Previously, the anthrax toxins activator ( atxA) mRNA had been used in our laboratory for the development of a biosensor for the detection of a single B . anthracis spore within 12 h . Changing the target sequence to the pag mRNA provided the ability to shorten the overall assay time significantly . The vaccine strain of B . anthracis (Sterne strain) was used in all experiments . A 500-microL sample containing as few as ten spores was mixed with 500 microL growth medium and incubated for 30 min for spore germination and mRNA production . Thus, only spores that are viable were detected . Subsequently, RNA was extracted from lysed cells, selectively amplified using NASBA, and rapidly identified by the biosensor . While the biosensor assay requires only 15 min assay time, the overall process takes 4 h for detection of ten viable B . anthracis spores, and is shortened significantly if more spores are present . The biosensor is based on an oligonucleotide sandwich-hybridization assay format . It uses a membrane flow-through system with an immobilized DNA probe that hybridizes with the target sequence . Signal amplification is provided when the target sequence hybridizes to a second DNA probe that has been coupled to liposomes encapsulating the dye sulforhodamine B . The amount of liposomes captured in the detection zone can be read visually or quantified with a hand-held reflectometer . The biosensor can detect as little as 1 fmol target mRNA (1 nmol L(-1)) . Specificity analysis revealed no cross-reactivity with 11 organisms tested, among them closely related species such as B . cereus, B . megaterium, B . subtilis, B . thuringiensis, Lactococcus lactis, Lactobacillus plantarum, and Chlostridium butyricum . Also, no false positive signals were obtained from nonviable spores . We suggest that this inexpensive biosensor is a viable option for rapid, on-site analysis providing highly specific data on the presence of viable B . anthracis spores.

Scand J Infect Dis, 2004, 36(6-7), 501 - 3
Lactobacillus gasseri causing Fournier's gangrene; Tleyjeh IM et al.; Fournier's gangrene is characterized as an aggressive necrotizing process caused by a polymicrobial infection that includes virulent organisms . We report the first case of Fournier's gangrene caused by a low-virulence organism, Lactobacillus gasseri.

Eur J Immunol, 2004 Sep, 34(9), 2520 - 8
Lactobacillus casei reduces CD8+ T cell-mediated skin inflammation; Chapat L et al.; Probiotics, including Lactobacilli, have been postulated to alleviate allergic and inflammatory diseases, but evidence that they exert an anti-inflammatory effect by immune modulation of pathogenic T cell effectors is still lacking . The aim of this study was to examine whether L . casei could affect antigen-specific T cell-mediated skin inflammation . To this end, we used contact hypersensitivity to the hapten 2,4-dinitrofluorobenzene, a model of allergic contact dermatitis mediated by CD8+ CTL and controlled by CD4+ regulatory T cells . Daily oral administration of fermented milk containing L . casei or L . casei alone decreased skin inflammation by inhibiting the priming/expansion of hapten-specific IFN-gamma-producing CD8+ effector T cells . The down-regulatory effect of the probiotics required the presence of CD4+ T cells, which control the size of the hapten-specific CD8+ T cell pool primed by skin sensitization . L . casei cell wall was as efficient as live L . casei to regulate both the CHS response and the hapten-specific CD8+ T cell response, suggesting that cell wall components contribute to the immunomodulatory effect of L . casei . This study provides the first evidence that oral administration of L . casei can reduce antigen-specific skin inflammation by controlling the size of the CD8+ effector pool .

J Med Food, 2004 Summer, 7(2), 223 - 8
Nucleic acid-based diagnosis of bacterial vaginosis and improved management using probiotic lactobacilli; Reid G et al.; Bacterial vaginosis (BV) is a common condition in women that represents an imbalance of the vaginal microflora, lactobacilli depletion, and excess growth of mainly anaerobic Gram-negative pathogens . Diagnosis is made using a series of tests or a Gram stain of a vaginal smear . Treatment with antibiotics is quite effective, but recurrences are common . A study of 55 vaginal samples from 11 postmenopausal women showed the presence of BV by the Gram stain-based Nugent scoring system, and polymerase chain reaction-denaturing gradient gel electrophoresis showed that Bacteroides or Prevotella species were the most common isolates recovered (24 of 25), with Escherichia coli, Staphylococcus aureus, and Streptococcus agalactiae also found in some samples . In one case, only Gardnerella vaginalis was found . These findings illustrate that BV remains common even among otherwise healthy women, but it is not caused solely by either Gardnerella or Mobiluncus . Use of a FemExam system (Cooper Surgical, Shelton, CT), based upon elevated pH and trimethylamine levels, to screen vaginal smears from 59 healthy women showed poor correlation with the Gram stain method . A randomized, placebo-controlled trial of these subjects showed that the lactobacilli-dominant microbiota was restored in subjects with BV but not in controls, following 2 months of daily oral intake of Lactobacillus rhamnosus GR-1 and Lactobacillus fermentum RC-14 . These studies show that nucleic acid-based methods are effective at identifying bacteria responsible for BV . If such methods could be used to develop a commercially available, self-use kit, women would be much better placed to take control of their own health, for example, using medicinal food or dietary supplement products such as the clinically proven probiotic strains L . rhamnosus GR-1 and L . fermentum RC-14.

Appl Environ Microbiol, 2004 Aug, 70(8), 4814 - 20
Overexpression of Lactobacillus casei D-hydroxyisocaproic acid dehydrogenase in cheddar cheese; Broadbent JR et al.; Metabolism of aromatic amino acids by lactic acid bacteria is an important source of off-flavor compounds in Cheddar cheese . Previous work has shown that alpha-keto acids produced from Trp, Tyr, and Phe by aminotransferase enzymes are chemically labile and may degrade spontaneously into a variety of off-flavor compounds . However, dairy lactobacilli can convert unstable alpha-keto acids to more-stable alpha-hydroxy acids via the action of alpha-keto acid dehydrogenases such as d-hydroxyisocaproic acid dehydrogenase . To further characterize the role of this enzyme in cheese flavor, the Lactobacillus casei d-hydroxyisocaproic acid dehydrogenase gene was cloned into the high-copy-number vector pTRKH2 and transformed into L . casei ATCC 334 . Enzyme assays confirmed that alpha-keto acid dehydrogenase activity was significantly higher in pTRKH2:dhic transformants than in wild-type cells . Reduced-fat Cheddar cheeses were made with Lactococcus lactis starter only, starter plus L . casei ATCC 334, and starter plus L . casei ATCC 334 transformed with pTRKH2:dhic . After 3 months of aging, the cheese chemistry and flavor attributes were evaluated instrumentally by gas chromatography-mass spectrometry and by descriptive sensory analysis . The culture system used significantly affected the concentrations of various ketones, aldehydes, alcohols, and esters and one sulfur compound in cheese . Results further indicated that enhanced expression of d-hydroxyisocaproic acid dehydrogenase suppressed spontaneous degradation of alpha-keto acids, but sensory work indicated that this effect retarded cheese flavor development.

Appl Environ Microbiol, 2004 Aug, 70(8), 4807 - 13
Effects of different spices used in production of fermented sausages on growth of and curvacin A production by Lactobacillus curvatus LTH 1174; Verluyten J et al.; Lactobacillus curvatus LTH 1174, a fermented sausage isolate, produces the listericidal bacteriocin curvacin A . The effect of different spices relevant for the production of fermented sausages was investigated in vitro through laboratory fermentations with a meat simulation medium and an imposed pH profile relevant for Belgian-type fermented sausages . The influence on the growth characteristics and especially on the kinetics of curvacin A production with L . curvatus LTH 1174 was evaluated . Pepper, nutmeg, rosemary, mace, and garlic all decreased the maximum specific growth rate, while paprika was the only spice that increased it . The effect on the lag phase was minor except for nutmeg and especially for garlic, which increased it, yet garlic was stimulatory for biomass production . The maximum attainable biomass concentration (X(max)) was severely decreased by the addition of 0.40% (wt/vol) nutmeg, while 0.35% (wt/vol) garlic or 0.80% (wt/vol) white pepper increased X(max) . Nutmeg decreased both growth and bacteriocin production considerably . Garlic was the only spice enhancing specific bacteriocin production, resulting in higher bacteriocin activity in the cell-free culture supernatant . Finally, lactic acid production was stimulated by the addition of pepper, and this was not due to the manganese present because an amount of manganese that was not growth limiting was added to the growth medium . Addition of spices to the sausage mixture is clearly a factor that will influence the effectiveness of bacteriocinogenic starter cultures in fermented-sausage manufacturing.

J Bacteriol, 2004 Aug, 186(16), 5529 - 32
Characterization of lipoteichoic acids as Lactobacillus delbrueckii phage receptor components; Raisanen L et al.; Lipoteichoic acids (LTAs) were purified from Lactobacillus delbrueckii subsp . lactis ATCC 15808 and its LL-H adsorption-resistant mutant, Ads-5, by hydrophobic interaction chromatography . L . delbrueckii phages (LL-H, the LL-H host range mutant, and JCL1032) were inactivated by these poly(glycerophosphate) type of LTAs in vitro in accordance to their adsorption to intact ATCC 15808 and Ads-5 cells.

J Agric Food Chem, 2004 Aug 11, 52(16), 5233 - 9
Evaluation of vinification lees as a general medium for lactobacillus strains; Bustos G et al.; Lactobacillus species present high nutritional requirements, so it is necessary to find new low-cost nutrient components for fermentation media . This work compares the utilization of vinification lees (an important residue of wineries) from red and white winemaking technology, distilled or not . An amount of 20 g of lees/L was used as the unique nutrient to obtain lactic acid from glucose using Lactobacillus strains with different properties: L . plantarum CECT-221, L . pentosus CECT-4023, L . casei CECT-5275, and L . coryniformis subsp . torquens CECT-25600 . Only L . casei using distilled lees showed values (Pmax = 92.1 g/L and Y(P/S) = 1.05 g/g) similar to those obtained with the MRS broth . The UV spectra of white and red lees, distilled or not, allowed an interpretation of the different phenolic compounds present and their influence on the fermentation . Their detoxification by extraction with organic compounds and fermentation with L . pentosus was also considered . Time courses of glucose and lactic acid were modeled according to reported models to obtain more information about the process.

Appl Microbiol Biotechnol . 2004 Jul 29; {Epub ahead of print}
Production of l-lactic acid from a mixture of xylose and glucose by co-cultivation of lactic acid bacteria; Taniguchi M et al.; The production of optically pure lactic acid in a high yield from xylose or a mixture of xylose and glucose, which is a model hydrolysate of lignocellulose, is described . In a single cultivation, Enterococcus casseliflavus produced 38 g/l of lactic acid with an optical purity of 96% enantiomeric excess (ee) and 6.4 g/l of acetic acid from 50 g/l of xylose when MRS medium was used . When a mixture of 50 g/l of xylose and 100 g/l of glucose was used as the carbon source in a cultivation of E . casseliflavus alone, glucose was converted to lactic acid in the early phase of the cultivation but xylose was hardly consumed . In a co-cultivation where E . casseliflavus and Lactobacillus casei specific for glucose were simultaneously inoculated, little or no lactic acid was produced after the glucose was almost consumed . A co-cultivation with two-stage inoculation (in which E . casseliflavus was added at a cultivation time of 40 h after L . casei cells were inoculated) resulted in complete consumption of 50 g/l of xylose and 100 g/l of glucose . In the co-cultivation, 95 g/l of lactic acid with a high optical purity of 96% ee was obtained at 192 h . Such a co-cultivation using two microorganisms specific for each sugar is considered to be one promising cultivation technique for the efficient production of lactic acid from a sugar mixture derived from lignocellulose.

Microbiology, 2004 Aug, 150(Pt 8), 2565 - 73
Characterization of vaginal microbial communities in adult healthy women using cultivation-independent methods; Zhou X et al.; The normal microbial flora of the vagina plays an important role in preventing genital and urinary tract infections in women . Thus an accurate understanding of the composition and ecology of the ecosystem is important to understanding the aetiology of these diseases . Common wisdom is that lactobacilli dominate the normal vaginal microflora of post-pubertal women . However, this conclusion is based on methods that require cultivation of microbial populations; an approach that is known to yield a biased and incomplete assessment of microbial community structure . In this study cultivation-independent methods were used to analyse samples collected from the mid-vagina of five normal healthy Caucasian women between the ages of 28 and 44 . Total microbial community DNA was isolated following resuspension of microbial cells from vaginal swabs . To identify the constituent numerically dominant populations in each community 16S rRNA gene libraries were prepared following PCR amplification using the 8f and 926r primers . From each library, the DNA sequences of approximately 200 16S rRNA clones were determined and subjected to phylogenetic analyses . The diversity and kinds of organisms that comprise the vaginal microbial community varied among women . Species of Lactobacillus appeared to dominate the communities in four of the five women . However, the community of one woman was dominated by Atopobium sp., whereas a second woman had appreciable numbers of Megasphaera sp., Atopobium sp . and Leptotrichia sp., none of which have previously been shown to be common members of the vaginal ecosystem . Of the women whose communities were dominated by lactobacilli, there were two distinct clusters, each of which consisted of a single species . One class consisted of two women with genetically divergent clones that were related to Lactobacillus crispatus, whereas the second group of two women had clones of Lactobacillus iners that were highly related to a single phylotype . These surprising results suggest that culture-independent methods can provide new insights into the diversity of bacterial species found in the human vagina, and this information could prove to be pivotal in understanding risk factors for various infectious diseases.

Arch Pediatr, 2004 Aug, 11(8), 903 - 7
{Treatment of acute diarrhea: prescription patterns by private practice pediatricians}; Uhlen S et al.; Acute gastroenteritis remains a frequent illness in infants and children with still important morbidity and mortality rates . Oral rehydratation solutions (ORS) and early refeeding are the main recommendations . Indication of drugs remains limited . OBJECTIVE: To evaluate the management of acute diarrhea by private practice pediatricians of France . METHODS: A questionnaire concerning ORS, dietary formula, antidiarrheal diet, antibiotherapy, antidiarrheal drugs was sent to all 2907 private pediatricians of France . RESULTS: Six hundred twenty-nine questionnaires were analyzed (22%) . Three hundred and ninety-seven pediatricians (63%) prescribed systematically an ORS, 294 (47%) changed formula, 412 (66%) prescribed a regimen . Antibiotic was prescribed after coproculture (81%), when glairy and bloody diarrhea (65%), associated infectious disease (63%), toxi-infectious syndrome (42%) or immunodeficiency were present (28%) . Most pediatricians (97%) prescribed at least one drug: diosmectite (84%), Lactobacillus acidophilus (63%), Saccharomyces boulardii (62%), racecadotril (62%), loperamide (28%), attapulgite de Mormoiron (26%), nifuroxazide (20%) . Drugs were prescribed more often for their effectiveness than for comfort . CONCLUSION: This study demonstrates the discrepancies that remain between recommendations and practical care in the treatment of acute diarrhea in children . Private French pediatricians often prescribe drugs.

Lett Appl Microbiol, 2004, 39(3), 284 - 9
Barotolerance is inducible by preincubation under hydrostatic pressure, cold-, osmotic- and acid-stress conditions in Lactobacillus sanfranciscensis DSM 20451T; Scheyhing CH et al.; AIMS: This study addresses the inducibility of barotolerance by preincubation of Lactobacillus sanfranciscensis DSM 20451T under various sublethal stress conditions . METHODS AND RESULTS: Stress conditions which reduce the growth rate of L . sanfranciscensis DSM 20451T to 10% of its maximum were determined . These conditions were met at 43, 12.5 degrees C, a pH value of 3.7, 1.9% NaCl, or 80 MPa respectively . In contrast to heat preincubation, other prestresses, including salt, cold and pressure led to an increase of barotolerance by hydrostatic pressure of 300 MPa for 30 min . Stationary-phase cells also showed an increased barotolerance . Sublethal pressure leads to enhanced heat tolerance . CONCLUSIONS: Stress response to salt, low temperature and acidic pH as well as starvation overlap with that one to high pressure by inducing barotolerance . SIGNIFICANCE AND IMPACT OF THE STUDY: Inactivation of bacteria by high pressure treatment is influenced by their history which modulates barotolerance . Mechanisms of barotolerance appear different from heat shock defence.

Lett Appl Microbiol, 2004, 39(3), 267 - 73
Development of a 16S rDNA-targeted PCR assay for monitoring of Lactobacillus plantarum and Lact . rhamnosus during co-cultivation for production of inoculants for silages; Klocke M et al.; AIMS: This paper reports a simple, rapid approach for the parallel detection of Lactobacillus plantarum and Lact . rhamnosus in co-culture in order to produce an inoculant mixture for silage purposes . METHODS AND RESULTS: The 16S rDNA-targeted PCR primers were established for parallel detection of Lact . plantarum and Lact . rhamnosus in a single multiplex PCR . A protocol for application of these primers in direct PCR as well as colony-direct (CD) PCR was developed . These primers were also applicable for the estimation of the relative amount of each DNA type in mixed probes (semi-quantitative PCR) . CONCLUSIONS: The PCR assay presented in this study is a robust, fast and semi-quantitative approach for detection of Lact . plantarum and Lact . rhamnosus in liquid cultures as well as on agar plates . SIGNIFICANCE AND IMPACT OF THE STUDY: This method provides an effective tool for the establishment of a regime for co-cultivation of Lact . plantarum and Lact . rhamnosus . This would enable faster and thus cost-reduced production of ensiling inoculants.

Lett Appl Microbiol, 2004, 39(3), 240 - 5
Comparative analysis of conserved genetic markers and adjacent DNA regions identified in beer-spoilage lactic acid bacteria; Suzuki K et al.; AIMS: To conduct an inter-species comparative study on the nucleotide sequences of the conserved DNA regions surrounding ORF5, a genetic marker for differentiating beer-spoilage lactic acid bacteria . METHODS AND RESULTS: The conserved DNA regions surrounding ORF5 were examined by PCR analysis, using three beer-spoilage strains, Lactobacillus brevis ABBC45C, L . paracollinoides LA2T and Pediococcus damnosus ABBC478 . As a result, the DNA regions containing ORF1-7, originally found in ABBC45C, appeared to be conserved among the three strains, while the downstream region was not found in L . paracollinoides LA2T and P . damnosus ABBC478 . The sequencing analysis of the conserved DNA regions of LA2T and ABBC478 revealed ca 99% nucleotide sequence identities with that of ABBC45C . CONCLUSIONS: The nucleotide sequences of the ca 8.2 kb DNA regions containing ORF1-7 were virtually identical among the three strains belonging to different species . The internal organizations of the ORFs were found to be remarkably similar . SIGNIFICANCE AND IMPACT OF THE STUDY: The level of nucleotide sequence identities suggests the DNA regions surrounding ORF5 were horizontally acquired by these beer-spoilage strains belonging to the three different species of lactic acid bacteria.

Biotechnol Bioeng, 2004 Aug 20, 87(4), 525 - 36
A novel convenient procedure for extractive work-up of whole-cell biotransformations using de-emulsifying hydrolases; Jorg G et al.; Extractive work-up of whole-cell biotransformations generally suffers from the formation of stable gels and slimes upon addition of the organic solvent to the cell suspension and the cell-free solution, respectively . This problem has been overcome by enzymatic lysis of emulsifying agents present in the medium through addition of hydrolases . Of these agents, proteases have exhibited the most powerful de-emulsifying activity . Enzyme treatment of cell-free culture media of Saccharomyces cerevisiae with pronase E drastically reduced phase separation time (t(p)) from 1 week to 30 min without significantly affecting product integrity . Yeast cell suspensions were de-emulsified best with protease N-01, where phase separation was complete after 1 h . As was exemplified with cell-free culture media of Lactobacillus kefir, wherein addition of pronase E or protease N-01 reduced t(p) from 1 week to 2 h each, this practical, ready-to-use method is appropriate for both fungal and bacterial biocatalysts.

Nahrung, 2004 Jun, 48(3), 218 - 20
Metabolic activities of Lactobacillus spp . strains isolated from kefir; Yuksekdag ZN et al.; A total of 21 strains of Lactobacillus species were isolated from Turkish kefir samples, in order to select the most suitable strains according to their metabolic activities including probiotic properties . As a result of the identification tests, 21 Lactobacillus isolates were identified as L . acidophilus (4%), L . helveticus (9%), L . brevis (9%), L . bulgaricus (14%), L . plantarum (14%), L . casei (19%) and L . lactis (28%) . The amount of produced lactic acid, hydrogen peroxide, proteolytic activity, and acetaldehyde productions of Lactobacillus spp . were determined . Different amounts of lactic acid were produced by strains studies; however, lactic acid levels were 1.7-11.4 mg/mL . All strains produced hydrogen peroxide . L . bulgaricus Z14L strain showed no proteolytic activity, L . casei Z6L strain produced the maximum amount (0.16 mg/mL) of proteolytic activity . Acetaldehyde concentration produced in Lactobacillus strains ranged between 0.88-3.52 microg/mL.

J Nutr, 2004 Aug, 134(8), 1964 - 9
Lactobacillus GG bacteria ameliorate arthritis in Lewis rats; Baharav E et al.; Probiotic bacteria have beneficial effects in infectious and inflammatory diseases, principally in bowel disorders . In the case of chronic progressive autoimmune arthritides, a major goal of treatment is to reduce inflammation . We hypothesized that probiotic bacteria would ameliorate inflammation found in arthritis models . To assess this effect, Lewis rats were injected with 50 microg bovine alpha-tropomyosin (TRM) or complete Freund's adjuvant (CFA) to induce tropomyosin arthritis (TA) or adjuvant arthritis (AA), respectively . In both models, the rats were divided into 6 groups and fed 0.5 mL/d of the following suspensions: 1) heat-killed Lactobacillus GG (LGG) bacteria; 2) live LGG, both 10(11) colony-forming units (cfu)/L; 3) sterilized milk; 4) plain yogurt; 5) yogurt containing 10(11) cfu/L LGG; or 6) sterilized water . In the disease-prevention experiments, feeding started 1 wk before or after disease induction . In the therapeutic experiments, feeding was initiated at the onset of clinical arthritis . In all experiments, there were significant interactions between time and treatment (P < 0.001), except for milk, which had no effect in the therapeutic experiment . Histologically, rats fed yogurt containing LGG had a milder inflammation in all experiments (P < 0.05), whereas rats fed plain yogurt exhibited a moderate inflammatory score only in the prevention experiments . Anti-TRM antibody titers were not affected by any of the treatments in any of the experiments . Ingestion of live or heat-killed human LGG had a clinically beneficial effect on experimental arthritis . Our observation of the remarkable preventive and curative effect on arthritis using commercial yogurts containing lactobacilli, especially LGG, suggests the need for investigation of these agents in arthritic patients.

Avian Dis, 2004 Apr-Jun, 48(2), 279 - 86
Salmonella enterica serovar typhimurium colonization of the crop in the domestic turkey: influence of probiotic and prebiotic treatment (Lactobacillus acidophilus and lactose); Johannsen SA et al.; Acute colonization of the crop of the domestic turkey by Salmonella enterica serovar typhimurium (ST) was examined . The influences of preharvest probiotic and prebiotic treatment with lactobaccilli and lactose on crop colonization with ST were also investigated . Prior to Salmonella challenge, poults received 2.5% lactose and Lactobacillus acidophilus (1.9 x 10(9) organisms/liter) in the only source of drinking water from 1 day old to termination . At 3-wk-old, turkey poults were challenged with ST (1.7 X 10(8) colony-forming units {CFU}/ml) before their natural nocturnal fast to determine the potential effects of supplementation on crop colonization when the crop was engorged and subsequently undergoing emptying . Crop ingesta and tissue were collected at time points 30 min and 4, 8, and 24 hr postchallenge and ST levels were determined . High levels of ST were detected in the crop . For instance, for the poults not receiving lactose or lactobacilli, 30 min after ST challenge, there were 4.4 x 10(7) CFU in the crop ingesta and 5.3 x 10(5) CFU in the crop wall . Ingesta ST levels dropped dramatically to 1.0 x 10(6) CFU after 4 hr as the crop emptied . Crop wall ST levels were steady during the nocturnal crop evacuation . Immunohistochemical staining demonstrated ST in close association with the crop epithelium . Treatment with lactose and L . acidophilus supplementation did not reduce ST colonization.

Int J Food Microbiol, 2004 Sep 1, 95(2), 219 - 23
Lactobacillus reuteri in bovine milk fermented decreases the oral carriage of mutans streptococci; Nikawa H et al.; The effect of Lactobacillus reuteri against one of the major cariogenic organism, Streptococcus mutans, was studied . Yogurt products containing L.reuteri showed a significant growth inhibitory effect against S . mutans, whilst yoghurts with lactobaccilli other than L . reuteri did not show such inhibition . Further, double-blind, placebo-controlled trial demonstrated that consuming yogurt with L . reuteri significantly reduced the oral carriage of mutans streptococci, compared with the placebo yogurt.

Int J Food Microbiol, 2004 Sep 1, 95(2), 157 - 68
Application of central composite design and response surface methodology to the fermentation of olive juice by Lactobacillus plantarum and Debaryomyces hansenii; Tsapatsaris S et al.; The objectives of this study were to investigate the effects of NaCl, calcium acetate and calcium lactate in concentrations corresponding to ionic strengths equivalent to 2-10%, w/v salt brines as well as the 50% replacement of NaCl contained in the above mixture by KCl . A central composite design and response surface methodology were used to optimize the maximum specific growth rate of Lactobacillus plantarum ATCC 8014 and Debaryomyces hansenii 2114 . The fermentation was carried out in olive juice obtained from Kalamon black olives at 30 degrees C with initial pH 5.0 . Mathematical models describing the combined effects of these factors on the maximum specific growth rate of L . plantarum or D . hansenii were established . Both strains in single cultures showed higher maximum specific growth rate in olive juice supplemented with NaCl/KCl, Ca-acetate and Ca-lactate . But in mixed culture fermentations of olive juice supplemented with NaCl, Ca-acetate and Ca-lactate, higher specific growth rates were obtained . Under the optimum growth conditions determined for the single culture fermentations, i.e . 378.4 mM NaCl, 34.1 mM Ca-acetate and 39.9 mM Ca-lactate, mixed culture fermentation was undertaken by varying the time of inoculation of the yeast strain . When D . hansenii was inoculated 48 h before L . plantarum the maximum specific growth rate of L . plantarum was increased to 0.247 per hour, which was significantly higher compared to L . plantarum alone (0.211 per hour) . In mixed culture fermentation of olive juice supplemented with the mixture of NaCl/KCl under similar conditions as above, a maximum specific growth rate of L . plantarum of 0.218 per hour was determined . The optimum conditions determined for mixed culture fermentation are useful in fermentation of black olives Kalamon variety under lower salt content.

Int J Food Microbiol, 2004 Sep 1, 95(2), 127 - 35
Expression of the catalase gene katA in starter culture Lactobacillus plantarum TISTR850 tolerates oxidative stress and reduces lipid oxidation in fermented meat product; Noonpakdee W et al.; The catalase gene katA of Lactobacillus sakei SR911 was cloned and expressed in Escherichia coli UM2 and Lactobacillus plantarum TISTR850 under strong lactococcal promoter P59 in E . coli-lactococcus expression vector pIL1020 . The L . plantarum TISTR850 is a catalase-deficient strain isolated from local fermented meat product . The recombinant L . plantarum TISTR850 was shown to decompose hydrogen peroxide, and catalase activity approximately three times higher that of natural catalase-producing strain L . sakei SR911 . The recombinant protein was also detected by in situ activity staining of the catalase enzyme . The recombinant L . plantarum TISTR850 did not accumulate hydrogen peroxide under glucose-limited aerobic conditions and remained viable after 60 h of incubation . The recombinant and host strain L . plantarum TISTR850 were used as starter cultures in the fermented meat product, and lipid oxidation was monitored over a 7-day storage at 20 degrees C determined as thiobarbituric acid-reactive substances (TBARS) value . The lipid oxidation level in the fermented meat product seeded with the catalase genetically modified starter culture L . plantarum TISTR850 was significantly lower than that of the natural catalase-deficient strain.

Int J Syst Evol Microbiol, 2004 Jul, 54(Pt 4), 1365 - 8
Lactobacillus saerimneri sp . nov., isolated from pig faeces; Pedersen C et al.; In studying the composition of the Lactobacillus flora of faeces from pigs fed different diets, isolates with notable differences in their 16S rRNA gene sequence compared to recognized species were found . Phenotypic characteristics together with 16S rRNA gene sequences revealed that the isolates represented a novel species belonging to the Lactobacillus mali subgroup of lactobacilli . The name Lactobacillus saerimneri sp . nov . is proposed (type strain GDA154T=LMG 22087T=DSM 16049T=CCUG 48462T).

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