Plasmid, 2003 May, 49(3), 253 - 68 Three new insertion sequence elements ISLdl2, ISLdl3, and ISLdl4 in Lactobacillus delbrueckii: isolation, molecular characterization, and potential use for strain identification; Ravin V et al.; A group of new insertion sequence (IS) elements, ISLdl2, ISLdl3, and ISLdl4, from Lactobacillus delbrueckii subsp . lactis ATCC 15808 was isolated, characterized, and used for strain identification together with ISLdl1, recently characterized as an L . delbrueckii IS element belonging to the ISL3 family . ISLdl2 was 1367 bp in size and had a 24 bp IR and an 8 bp DR . The single ORF of ISLdl2 encoded a protein of 392 aa similar to transposases of the IS256 family . ISLdl3 had a single ORF encoding a protein of 343 aa similar to transposases of the IS30 family . Finally, ISLdl4 had a single ORF encoding a protein of 406 aa and displayed homology to the transposases of the IS110 family . ISLdl4 was only slight different from ISL4 (Accession No . AY040213) . ISLdl1, ISLdl2, and ISLdl4 were present in all of the 10 L . delbrueckii subsp . lactis and subsp . delbrueckii strains tested, as well as in three of the 11 L . delbrueckii subsp . bulgaricus strains tested . ISLdl3 was present only in four closely related strains of L . delbrueckii subsp . lactis . These IS elements were not observed in Lactobacillus rhamnosus, Lactobacillus acidophilus, Lactobacillus helveticus, or Lactobacillus plantarum . A cluster of IS elements, ISLdl1, ISLdl2, ISLdl3, ISLdl4, and ISL6, was observed in L . delbrueckii subsp . lactis strain ATCC 15808 . Within this cluster, ISLdl4 was inserted into ISLdl1 between the left IR and the start codon of ORF455, encoding a putative transposase . Most of the integration sites of the IS elements were strain-specific . We have observed that IS elements can migrate from one strain to another as integral parts of bacterial DNA by using phage LL-H as a vehicle . We demonstrate for the first time that inverse PCR and vectorette PCR methods with primers based on sequences of the IS elements could be used for identification of L . delbrueckii strains. Avian Pathol, 2003 Apr, 32(2), 173 - 81 Passage of Salmonella through the crop and gizzard of broiler chickens fed with fermented liquid feed; Heres L et al.; In vivo experiments were conducted in order to investigate the passage and bacterial reduction of Salmonella in the crop and gizzard of chickens when fed two different feeds . The chickens were fed dry conventional feed and fermented liquid feed . The fermented feed contains a relatively high concentration of lactic and acetic acid and lactobacilli . One and three week old broiler chickens were necropsied at short intervals after inoculation with Salmonella Enteritidis . Counts of Salmonella from the crop, gizzard, duodenum, caecum and colon/rectum were obtained . This revealed a sharper decrease of Salmonella in the anterior parts of the gastro-intestinal tract in chickens fed with fermented feed than in chickens fed dry feed . It is therefore concluded that fermented feed improves the barrier formed by the crop and gizzard . The reduction of Salmonella is fully realised in the crop and gizzard . The lower intestinal compartment did not show a substantial effect on the reduction of Salmonella . The performed in vivo method appeared to be an appropriate way to study intervention strategies that aim to control Salmonella by improving the barrier function of the upper gastro-intestinal tract. J Agric Food Chem, 2003 May 21, 51(11), 3287 - 93 Purification and structural characterization of 3-hydroxypropionaldehyde and its derivatives; Vollenweider S et al.; The compound 3-hydroxypropionaldehyde (3-HPA), together with HPA hydrate and HPA dimer, in aqueous solution forms a system with interesting chemical properties . Therefore, 3-HPA has attracted attention by the chemical industry for use as a precursor in the production of plastics, acrylic acid, and 1,3-propanediol and by the food industry, in using 3-HPA-producing Lactobacillus reuteri as a probiotic . To produce 3-HPA in high yield from glycerol, L . reuteri was used as a biotransformation system . A convenient chromatographic purification method was developed, and purified 3-HPA was analyzed using electrospray ionization mass spectrometry and (13)C NMR . Quantitative (13)C NMR revealed a concentration-dependent distribution of the three compounds forming the HPA system . At concentrations above 1.4 M, the HPA dimer was predominant . However, at concentrations relevant for biological systems, HPA hydrate was the most abundant, followed by the aldehyde form . Our results indicate that the dimeric form with expected antibiotic properties should not be the active form. Folia Microbiol (Praha), 2003, 48(1), 56 - 8 Evidence for chromosomal determination of fungicidal activity in strains of Lactobacillus brevis and Lactobacillus fermentum isolated from fermented foods; Osuntoki AA et al.; The genetic basis of the fungicidal activity of strains of Lactobacillus brevis and L . fermentum isolated from indigenous fermented foods was determined . A 5.5-kb plasmid was isolated from L . brevis while L . Fermentum was found to harbor no plasmid . Plasmid curing indicated no correlation between the plasmid and the fungicidal activity of the Lactobacillus species . The fungicidal activity of the isolated organisms can be supposed to be mediated by the chromosome . No antibiotic resistance genetic markers were detected on the plasmid and hence it was classified as cryptic. Appl Microbiol Biotechnol, 2003 Oct, 62(5-6), 498 - 502 Epub 2003 May 13. Control of Lactobacillus contaminants in continuous fuel ethanol fermentations by constant or pulsed addition of penicillin G; Bayrock DP et al.; The addition of penicillin G to combat microbial contamination in continuous fuel alcohol fermentations was performed using both continuous and pulsed addition regimes . In continuous fermentations where both Saccharomyces cerevisiae and Lactobacillus paracasei were present, the mode of addition of penicillin G determined final numbers of viable L . paracasei . When the same overall average concentration of penicillin G was added in both pulsed and continuous modes, the initial viable number of L . paracasei (8.0 x 10(9) cfu ml(-1)) decreased to a greater degree (1.02 x 10(5) cfu ml(-1) L . paracasei) when penicillin G was pulsed at 6 h frequencies at an overall average concentration of 2,475 U/l than when penicillin G was added continuously at 2,475 U/l (2.77 x 10(5) cfu ml(-1) L . paracasei) . Pulsed additions over longer frequencies at 2,475 U/l were not as effective in reducing viable bacteria . Viable yeasts increased during both treatment conditions by more than 2-fold . The two addition regimes also eliminated the 40% decrease in ethanol concentration caused by the intentional bacterial infection . Although there was 3 times more bacterial death with 6 h pulsed additions compared to continuous additions of penicillin G at 2,475 U/l, there was, by that point, no practical difference in either final ethanol concentration or relative ethanol recovery. J Dairy Sci, 2003 Apr, 86(4), 1118 - 29 Intracellular esterase from Lactobacillus casei LILA: nucleotide sequencing, purification, and characterization; Fenster KM et al.; An esterase gene (estC) was isolated from a genomic library of Lactobacillus casei LILA . The estC gene consisted of a 777 bp open reading frame encoding a putative peptide of 28.9 kDa . A recombinant EstC fusion protein containing a C-terminal six-histidine tag was constructed and purified to electrophoretic homogeneity . Characterization of EstC revealed that it was a serine-dependent dimeric enzyme . Optimum temperature, NaCl concentration, and pH for EstC were determined to be 30 degrees C, 0% NaCl, and pH 5.5, respectively . EstC had significant activity under conditions simulating those of ripening cheese (10 degrees C, 4% NaCl, and pH 5.1) . Kinetic constants (KM and Vmax) were determined for EstC action on a variety of ethyl esters and ester compounds consisting of substituted phenyl alcohols and short n-chain fatty acids . For comparison purposes, the previously studied EstA from Lactococcus lactis MG1363 was purified to electrophoretic homogeneity and its substrate selectivity determined in a similar fashion . Different substrate selectivities were observed for EstC and EstA. Gut, 2003 Jun, 52(6), 827 - 33 Extracellular MUC3 mucin secretion follows adherence of Lactobacillus strains to intestinal epithelial cells in vitro; Mack DR et al.; BACKGROUND: Mucins are large complex glycoproteins that protect intestinal mucosal surfaces by limiting access of environmental matter to their epithelial cells . Several mucin genes have been described, including MUC3 that is a membrane associated mucin of the small intestine . Increased MUC3 mRNA transcription is induced by incubation of intestinal epithelial cells with a Lactobacillus strain known to be adherent to them . AIMS: To determine whether increased epithelial cell MUC3 mucin expression in response to Lactobacillus strains results in increased extracellular secretion of MUC3 mucins and the importance of epithelial cell adherence in modulation of MUC3 mucin expression . METHODS: HT29 cells grown to enhance expression of MUC3 mucins were incubated with selected Lactobacillus strains . Spent cell culture medium was collected for detection of secreted MUC3 mucins using dot blot immunoassay with a generated MUC3 antibody . Post-incubation HT29 cell RNA was collected for analysis of MUC3 expression by northern blot analysis using a MUC3 cDNA probe . In vitro binding studies using Lactobacillus strains incubated alone or coincubated with enteropathogenic Escherichia coli strain E2348/69 were used for adherence and inhibition of adherence studies, respectively . RESULTS: Lactobacillus strains with minimal ability to adhere to HT29 cells failed to induce upregulation of mucin gene expression . There was a direct correlation between upregulation of MUC3 mucin mRNA expression and extracellular secretion of MUC3 mucin . The same Lactobacillus strains that increased extracellular secretion of MUC3 mucin led to reduced adherence of enteropathogen E coli E2348/69 during coincubation experiments . CONCLUSION: Probiotic microbes induce MUC3 mucin transcription and translation with extracellular secretion of the MUC3 mucins . Epithelial cell adherence enhances the effects of probiotics on eukaryotic mucin expression. Microb Cell Fact . 2003 Apr 9;2(1):2. Cheese whey-induced high-cell-density production of recombinant proteins in Escherichia coli; Viitanen MI et al.; BACKGROUND: Use of lactose-rich concentrates from dairy processes for the induction of recombinant gene's expression has not received much attention although they are interesting low cost substrates for production of recombinant enzymes . Applicability of dairy waste for induction of recombinant genes in Escherichia coli was studied . Clones expressing Lactobacillus phage muramidase and Lactobacillus alcohol dehydrogenase were used for the experiments . RESULTS: Shake flask cultivations in mineral salt medium showed that cheese whey or deproteinised whey induced gene expression as efficiently as IPTG (isopropyl-beta-D-thiogalactopyranoside) or pure lactose . Addition of yeast extract or proteolytically degraded whey proteins did not improve the recombinant protein yield . In contrast, addition of yeast extract to the well-balanced mineral salt medium decreased the product yield . Feeding with glycerol provided sufficient amount of easily assimilable carbon source during the induction period without preventing lactose intake and induction by lactose . High-cell-density fed-batch cultivations showed that product yields comparable to IPTG-induction can be achieved by feeding bacteria with a mixture of glycerol and concentrated whey permeate during the induction . CONCLUSION: Whey and concentrated whey permeate can be applied as an alternative inducer in recombinant high-cell-density fed-batch fermentations . The yield of the recombinant product was comparable to fermentations induced by IPTG . In low-cell-density shake flask experiments the yield was higher with whey or whey permeate than with IPTG. Am J Gastroenterol, 2003 Apr, 98(4), 865 - 70 Effects of nonpathogenic bacteria on cytokine secretion by human intestinal mucosa; Borruel N et al.; OBJECTIVE: The human intestine harbors a complex microbial ecosystem, and the mucosa is the interface between the immune system and the luminal environment . The aim of this study was to elucidate whether host-bacteria interactions influence mucosal cytokine production . METHODS: Macroscopically normal colonic specimens were obtained at surgery from eight patients with neoplasm, and inflamed ileal specimens were obtained from two patients with Crohn's disease . Mucosal explants were cultured for 24 h with either nonpathogenic Escherichia coli ECOR-26, Lactobacillus casei DN-114 001, L . casei DN-114 056, L . casei ATCC-334, or Lactobacillus bulgaricus LB-10 . Each study included blank wells with no bacteria . Tissue and bacteria viability were confirmed by LDH release and culture . Concentration of tumor necrosis factor (TNF)alpha, transforming growth factor beta1, interleukin (IL)-8, and IL-10 was measured in supernatants . In parallel experiments, neutralizing anti-TNFalpha antibody was added to the culture . RESULTS: Co-culture of mucosa with bacteria did not modify LDH release . Co-culture with L . casei strains significantly reduced TNFalpha release, whereas E . coli increased it . These effects were observed both in normal and inflamed mucosa . In combination studies, L . casei DN-114 001 prevented TNFalpha stimulation by E . coli . L . casei DN-114 001 also reduced IL-8 release via a TNFalpha-independent pathway . L . casei DN-114 056 or E . coli increased IL-10 release in the presence of neutralizing anti-TNFalpha . CONCLUSIONS: Nonpathogenic bacteria interact with human intestinal mucosa and can induce changes in cytokine production that are strain specific. Br Poult Sci, 2003 Mar, 44(1), 139 - 44 Effects of Lactobacillus cultures on growth performance, abdominal fat deposition, serum lipids and weight of organs of broiler chickens; Kalavathy R et al.; 1 . The effects of a mixture of 12 Lactobacillus strains (LC) on the growth performance, abdominal fat deposition, serum lipids and weight of organs of broiler chickens were studied from 1 to 42 d of age . 2 . One hundred and thirty-six 1-d-old male broiler chicks were assigned at random to two dietary treatments: a basal diet (control), and a basal diet with 0.1% LC . 3 . The supplementation of LC in broiler diets improved the body weight gain and feed conversion rate from 1 to 42 d of age and was effective in reducing abdominal fat deposition but only after 28 d of age . 4 . The LC diets reduced serum total cholesterol, low density lipoprotein (LDL) cholesterol and triglycerides in broilers from 21 to 42 d of age . However, there was no significant difference in serum high density lipoprotein (HDL) cholesterol between control and LC-fed broilers . There was also no significant difference in the weights of organs of control and LC-fed broilers . 5 . The results indicated that the mixture of 12 Lactobacillus strains have a hypolipidaemic effect on broilers. Br Poult Sci, 2003 Mar, 44(1), 67 - 74 Effect of whole and demucilaged linseed in broiler chicken diets on digesta viscosity, nutrient utilisation and intestinal microflora; Alzueta C et al.; 1 . A study was undertaken to evaluate the effects of the inclusion of linseed or demucilaged linseed in the diet on the performance, fat and fatty acid digestibilities, metabolisability of gross energy and intestinal microflora . 2 . The dietary inclusions were 80 and 160 g/kg for linseed and 160 g/kg for demucilaged linseed . Diets were given to chickens from 1 to 23 d of age . 3 . Incorporation of linseed in the diet, particularly at 160 g/kg, depressed weight gain and food utilisation . Digestibility of fat and single fatty acids and dietary metabolisable energy were reduced . 4 . Inclusion of linseed in the diet markedly increased the viscosity of ileal digesta . Microbial activity in the ileum and caeca was not greatly affected, but there was a significant increase in the number of lactobacilli . 5 . The antinutritional effects caused by feeding linseed were partially overcome by substituting demucilaged linseed for linseed in the diet . 6 . Results from the current study suggest that the viscous properties of mucilage are a major factor in the observed antinutritional effects of linseed through increasing intestinal viscosity . Such effects might also be mediated by the gut microflora. J Clin Microbiol, 2003 May, 41(5), 2237 - 9 Fatal case of endocarditis due to Weissella confusa; Flaherty JD et al.; This is the first reported case of endocarditis due to the Lactobacillus-like vancomycin-resistant gram-positive bacillus Weissella confusa . Full identification and susceptibility testing of Lactobacillus-like organisms recovered in blood culture should be performed for patients with clinical presentations that suggest endocarditis. J Clin Microbiol, 2003 May, 41(5), 1925 - 8 BVBlue test for diagnosis of bacterial vaginosis; Myziuk L et al.; Bacterial vaginosis (BV) is a disorder of the vaginal ecosystem characterized by a shift in the vaginal flora from the normally predominant Lactobacillus to one dominated by sialidase enzyme-producing mixed flora . It is the most common cause of abnormal vaginal discharge in adult women . The BVBlue system (Gryphus Diagnostics, L.L.C.) is a chromogenic diagnostic test based on the presence of elevated sialidase enzyme in vaginal fluid samples . BVBlue was compared to the standard method for diagnosing BV (Amsel criteria and Nugent score) . Fifty-seven nonmenstruating women of > or =16 years of age who presented for a pelvic examination were recruited . Demographic features were collected via a self-administered questionnaire . The Amsel criteria were assessed based on three of four of the following characteristics of vaginal discharge: consistency, odor, pH, and presence of clue cells on Gram stain . BVBlue was compared to the Gram stain and Amsel criteria . The sensitivity, specificity, positive predictive value, and negative predictive value for BVBlue versus the Gram stain and Amsel criteria were 91.7, 97.8, 91.7, and 97.8% and 50.0, 100, 100, and 88.2%, respectively . A significantly greater proportion of patients with a vaginal pH of >4.5, a positive amine test, or with clue cells on vaginal Gram smear were found to have a positive BVBlue test (P < 0.001) . Women previously treated for BV were 2.98 times more likely to have another episode of BV . BVBlue is a useful point-of-care diagnostic tool to provide a presumptive diagnosis of BV, especially in situations where microscopic capabilities are unavailable. J Clin Microbiol, 2003 May, 41(5), 1881 - 7 DNA fingerprinting of Lactobacillus crispatus strain CTV-05 by repetitive element sequence-based PCR analysis in a pilot study of vaginal colonization; Antonio MA et al.; Lactobacillus crispatus is one of the predominant hydrogen peroxide (H(2)O(2))-producing species found in the vagina and is under development as a probiotic for the treatment of bacterial vaginosis . In this study, we assessed whether DNA fingerprinting by repetitive element sequence-based PCR (rep-PCR) can be used to distinguish the capsule strain of L . crispatus (CTV-05) from other endogenous strains as well as other species of vaginal lactobacilli . Vaginal and rectal lactobacilli were identified to the species level by using whole-chromosome probe DNA hybridization . The DNAs from L . crispatus, L . jensenii, L . gasseri, and an as-yet-unnamed H(2)O(2)-negative Lactobacillus species designated 1086V were subjected to rep-PCR . The results of gel electrophoresis and ethidium bromide staining of the DNA fingerprints obtained were compared . L . crispatus CTV-05 had a unique DNA fingerprint compared to all other lactobacilli . DNA fingerprints for 27 production lots of L . crispatus sampled from 1994 through 2001 were identical to that of the original strain isolated in 1993, suggesting strain stability . In a pilot study of nine women, this DNA fingerprinting method distinguished CTV-05 from other endogenous vaginal lactobacilli prior to and after vaginal capsule use . rep-PCR DNA fingerprinting is useful for strain typing and for evaluating longitudinal loss or acquisition of vaginal lactobacilli used as probiotics. Appl Environ Microbiol, 2003 May, 69(5), 3015 - 9 Unexpected thermal destruction of dried, glass bead-immobilized microorganisms as a function of water activity; Laroche C et al.; To help us understand the factors and mechanisms implicated in the death of microorganisms or their resistance to temperature in a low water activity environment, microorganisms were dried on the surface of glass beads before being subjected to high temperatures for a short period followed by rapid cooling . Two microorganisms were studied: the yeast Saccharomyces cerevisiae and the bacterium Lactobacillus plantarum . Experiments were carried out at 150, 200, and 250 degrees C, with four durations of heat treatment and seven levels of initial water activity between 0.10 and 0.70 . We observed an unexpected range of water activity, between 0.30 and 0.50, at which microorganisms were more resistant to the various treatments, with maximal viability at 0.35 for L . plantarum and 0.40 for S . cerevisiae. Appl Environ Microbiol, 2003 May, 69(5), 2674 - 83 Extent of genetic lesions of the arginine and pyrimidine biosynthetic pathways in Lactobacillus plantarum, L . paraplantarum, L . pentosus, and L . casei: prevalence of CO(2)-dependent auxotrophs and characterization of deficient arg genes in L . plantarum; Bringel F et al.; Lactic acid bacteria require rich media since, due to mutations in their biosynthetic genes, they are unable to synthesize numerous amino acids and nucleobases . Arginine biosynthesis and pyrimidine biosynthesis have a common intermediate, carbamoyl phosphate (CP), whose synthesis requires CO(2) . We investigated the extent of genetic lesions in both the arginine biosynthesis and pyrimidine biosynthesis pathways in a collection of lactobacilli, including 150 strains of Lactobacillus plantarum, 32 strains of L . pentosus, 15 strains of L . paraplantarum, and 10 strains of L . casei . The distribution of prototroph and auxotroph phenotypes varied between species . All L . casei strains, no L . paraplantarum strains, two L . pentosus strains, and seven L . plantarum strains required arginine for growth . Arginine auxotrophs were more frequently found in L . plantarum isolated from milk products than in L . plantarum isolated from fermented plant products or humans; association with dairy products might favor arginine auxotrophy . In L . plantarum the argCJBDF genes were functional in most strains, and when they were inactive, only one gene was mutated in more than one-half of the arginine auxotrophs . Random mutation may have generated these auxotrophs since different arg genes were inactivated (there were single point mutations in three auxotrophs and nonrevertible genetic lesions in four auxotrophs) . These data support the hypothesis that lactic acid bacteria evolve by progressively loosing unnecessary genes upon adaptation to specific habitats, with genome evolution towards cumulative DNA degeneration . Although auxotrophy for only uracil was found in one L . pentosus strain, a high CO(2) requirement (HCR) for arginine and pyrimidine was common; it was found in 74 of 207 Lactobacillus strains tested . These HCR auxotrophs may have had their CP cellular pool-related genes altered or deregulated. Wei Sheng Yan Jiu . 2003 Jan;32(1):56. {Lactobacillus casei microbiological assay of plasma and RBC concentrations of folic acid with 96-well microtiiter plates}; Huangfu Z et al.; To investigate the plasma and RBC concentrations of folic acid in normal persons . Microbiological assay was used to determine the plasma and RBC folic acid in 59 normal persons by using of 96-well microtiter plasma . The results showed that the plasma and RBC folio acid concentration were (8.2 +/- 2.9) microgram/L and (337.5 +/- 91.0) microgram/L in male and female normal persons (aged 18 to 32) respectively . The coefficient Variations determined within assay and between assay were 2.2% and 4.7% respectively . This method may reduce reagent costs, shortened the assay time, increase the sensitivity and improve the reproducibility . It can be used for large scale survey. Biosci Biotechnol Biochem, 2003 Feb, 67(2), 420 - 2 Effect of an oral administration of Lactobacillus casei strain Shirota on the natural killer activity of blood mononuclear cells in aged mice; Hori T et al.; The natural killer (NK) activity of blood mononuclear cells and splenocytes in aged mice fed on a diet containing Lactobacillus casei strain Shirota (LcS group) was significantly (P < 0.01) higher than that in mice fed on a diet without LcS . In the LcS group, there was a significant positive correlation (r = 0.63, P < 0.01) between the NK activity of blood mononuclear cells and the NK activity of splenocytes. Trends Biotechnol, 2003 May, 21(5), 217 - 23 Bacterial replacement therapy: adapting 'germ warfare' to infection prevention; Tagg JR et al.; The individual bacterial members of our indigeneous microbiota are actively engaged in an on-going battle to prevent colonisation and overgrowth of their terrain by competing microbes, some of which might have pathogenic potential for the host . Humans have long attempted to intervene in these bacterial interactions . Ingestion of probiotic bacteria, particularly lactobacilli, is commonly practiced to promote well-balanced intestinal microflora . As bacterial resistance to antimicrobials has increased, so too has research into colonisation of human tissues with specific effector strains capable of out-competing known bacterial pathogens . Recent progress is particularly evident in the application of avirulent Streptococcus mutans to the control of dental caries, alpha hemolytic streptococci to reduction of otitis media recurrences and Streptococcus salivarius to streptococcal pharyngitis prevention. Plasmid, 2003 Mar, 49(2), 118 - 29 Sequence and analysis of pBM02, a novel RCR cryptic plasmid from Lactococcus lactis subsp cremoris P8-2-47; Sanchez C et al.; This paper reports the complete nucleotide sequence of the 3.85 kbp plasmid pBM02 from Lactococcus lactis subsp . cremoris P8-2-47 . Analysis of the sequence predicted six ORFs larger than 25 amino acids . They all were transcribed from the same strand and organized in two functional cassettes: the replication region and a putative mobilization region . In the replication region, two ORFs specifying proteins homologous to others found in some classes of rolling circle-replicating plasmids were encountered (copG and repB) . In fact, single-stranded DNA was detected as a replication intermediate of pBM02 . copG and repB, together with some upstream sequences, formed part of the minimal replication unit of the plasmid . Interestingly, pBM02 shared a 212 bp stretch with plasmids of the pWV01 type, in which the whole single-strand origin of replication is included . In the mobilization region, an ORF coding for a mobilization-like protein was present, preceded by a putative oriT sequence homologous to that of plasmid pMV158 . The replicon of pBM02 is of the wide-host range type, and functions in both Gram-positive and Gram-negative bacteria, including Lactobacillus casei, Lactobacillus plantarum, Bacillus subtilis, and Escherichia coli. FEMS Microbiol Lett, 2003 Apr 25, 221(2), 285 - 91 Comparison of the thickening properties of four Lactobacillus delbrueckii subsp . bulgaricus strains and physicochemical characterization of their exopolysaccharides; Petry S et al.; It is now well established that physicochemical properties of exopolysaccharides (EPS) can vary between strains of a given species and according to growth conditions . The EPS production of four strains of Lactobacillus bulgaricus was monitored during growth in milk and in a chemically defined media . All strains, including the non-ropy one, produced EPS . The monosaccharide composition, molar mass (M(w)), and intrinsic viscosity of these EPS were determined and compared . Further characterization using high-performance size-exclusion chromatography revealed the presence of two fractions in all EPS: one fraction exhibited a high M(w) and a high intrinsic viscosity while the other had a low M(w) and a low intrinsic viscosity . Strikingly, the EPS synthesized by the non-ropy strain was mainly composed of the low-M(w) fraction while for the ropy strains, the fraction of high M(w) varied between 43 and 90% . According to our results, we propose that the ratio between the high-M(w) and low-M(w) fractions is critical for the texturing properties of L . bulgaricus EPS. Appl Biochem Biotechnol, 2003 Spring, 105 -108, 637 - 47 Production of lactic acid from food wastes; Kim KI et al.; Conversion of food wastes into lactic acid by simultaneous saccharification and fermentation (SSF) was investigated . The process involves saccharification of the starch component in food wastes by a commercial amylolytic enzyme preparation (a mixture of amyloglucosidase, alpha-amylase, and protease) and fermentation by Lactobacillus delbrueckii . The highest observed overall yield of lactic acid in the SSF was 91% of theoretical . Lactic acid concentration as high as 80 g/L was attainable in 48 h of the SSF . The optimum operating conditions for the maximum productivity were found to be 42 degrees C and pH 6.0 . Without supplementation of nitrogen-containing nutrients, the lactic acid yield in the SSF decreased to 60%: 27 g/L of lactic acid from 60 g/L of food waste . The overall performance of the SSF, however, was not significantly affected by the elimination of mineral supplements. Obstet Gynecol Surv, 2003 May, 58(5), 351 - 8 Common complementary and alternative therapies for yeast vaginitis and bacterial vaginosis: a systematic review; Van Kessel K et al.; This article is a systematic review of the literature regarding the most commonly used complementary and alternative medicine (CAM) therapies for yeast vaginitis and bacterial vaginosis . A search was conducted of all published literature on conventional search engines (PubMed, EMBASE, the Cochrane Registry, CINAHL, LILACS) and alternative medicine databases (Natural Medicines Comprehensive Database, Longwood Herbal Taskforce, and Alternative Medicine Alert), for all studies of the five most commonly used CAM treatments of vaginitis . Inconsistencies in definition of vaginitis, type of intervention, control groups, and outcomes prevented performance of a meta-analysis, and paucity of high-quality studies made ranking by evidence-based scales unsuitable . Lactobacillus recolonization (via yogurt or capsules) shows promise for the treatment of both yeast vaginitis and bacterial vaginosis with little potential for harm . Boric acid can be recommended to women with recurrent vulvovaginal Candidal infections who are resistant to conventional therapies, but can occasionally cause vaginal burning . Because of associated risks in the absence of well-documented clinical benefits, douching remains a practice that should not be recommended for the treatment of vaginitis . Finally, tea tree oil and garlic show some in vitro potential for the treatment of vaginitis, but the lack of in vivo studies preclude their recommendation to patients for the time-being . The available evidence for CAM treatments of vaginitis is of poor quality despite the prevalent use of these therapies . Well-designed randomized, controlled trials investigating the efficacy and safety of these therapies for vaginitis are needed before any reliable clinical recommendations can be made . TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians . LEARNING OBJECTIVES: After completion of this article, the reader will be able to list the most common complementary and alternative medicine therapies for vaginitis, summarize the data surrounding the efficacy of each therapy, describe the adverse affects of each therapy, and outline which therapies are recommended and not recommended for vaginitis. Protein Sci, 2003 May, 12(5), 1131 - 5 Peptidase family U34 belongs to the superfamily of N-terminal nucleophile hydrolases; Pei J et al.; Peptidase family U34 consists of enzymes with unclear catalytic mechanism, for instance, dipeptidase A from Lactobacillus helveticus . Using extensive sequence similarity searches, we infer that U34 family members are homologous to penicillin V acylases (PVA) and thus potentially adopt the N-terminal nucleophile (Ntn) hydrolase fold . Comparative sequence and structural analysis reveals a cysteine as the catalytic nucleophile as well as other conserved residues important for catalysis . The PVA/U34 family is variable in sequence and exhibits great diversity in substrate specificity, to include enzymes such as choloyglycine hydrolases, acid ceramidases, isopenicillin N acyltransferases, and a subgroup of eukaryotic proteins with unclear function. Z Naturforsch {C}, 2003 Mar-Apr, 58(3-4), 225 - 9 Effect of aeration on the production of carotenoid pigments by Rhodotorula rubra-lactobacillus casei subsp . casei co-cultures in whey ultrafiltrate; Simova ED et al.; Under intensive aeration (1.3 l/l min) the associated growth of Rhodotorula rubra GED2 and Lactobacillus casei subsp . casei in cheese whey ultrafiltrate (55 g lactose/l) proceeded effectively for both cultures with production of maximum carotenoids (12.4 mg/l culture fluid) . For maximum amount of carotenoids synthesized in the cell, the yeast required more intensive aeration than the aeration needed for synthesis of maximum concentration of dry cells . Maximum concentration of carotenoids in the cell (0.49 mg/g dry cells) was registered with air flow rate at 1.3 l/l min, and of dry cells (27.0 g/l) at 1.0 l/l min . An important characteristic of carotenogenesis by Rhodotorula rubra GED2 + Lactobacillus casei subsp . casei was established--the intensive aeration (above 1.0 l/l min) stimulated beta-carotene synthesis (60% of total carotenoids). Int J Syst Evol Microbiol, 2003 Mar, 53(Pt 2), 513 - 7 Lactobacillus versmoldensis sp . nov., isolated from raw fermented sausage; Krockel L et al.; Lactobacillus versmoldensis sp . nov . (KU-3T) was isolated from raw fermented sausages . The new species was present in high numbers, and frequently dominated the lactic acid bacteria (LAB) populations of the products . 16S rDNA sequence data revealed that the isolates are closely related to the species Lactobacillus kimchii DSM 13961T, Lactobacillus paralimentarius DSM 13238T, Lactobacillus alimentarius DSM 20249T and Lactobacillus farciminis DSM 20184T . DNA-DNA reassociation data, however, clearly distinguished the new isolates from these species; they showed a low degree of DNA relatedness with the type strains of this group of phylogenetically closely related lactobacilli . These results warrant separate species status for strain KU-3T, for which the name Lactobacillus versmoldensis sp . nov . is proposed . The type strain is KU-3T (=DSM 14857T =NCCB 100034T =ATCC BAA-478T). Poult Sci, 2003 Apr, 82(4), 657 - 63 Effects of isomalto-oligosaccharides on broiler performance and intestinal microflora; Zhang WF et al.; Dose effects of dietary isomalto-oligosacchrides (IMO) on broiler growth performance and characteristics of the intestinal microflora were compared . Three hundred sixty male broilers were randomly allotted to five treatments, with eight replicate pens per treatment and nine chicks per pen . Chicks were fed either a basal diet (control) or the basal diet plus 0.3, 0.6, 0.9, or 1.2% IMO . All chicks had access to feed and water ad libitum during the 7-wk experiment . At the end of the experiment, eight chicks per treatment were randomly chosen to measure the thymus index . Additionally, six birds per treatment were randomly selected to determine viable bacterial counts of Lactobacillus, Escherichia coli, and total aerobes in the digestive tract . The digesta of all the killed birds were also used to measure short-chain fatty acid (SCFA) levels . The results indicate that IMO enhanced growth performance during the initial 3 wk, but no further effects were detected during the latter 4 wk of the experiment . Isobutyrate level in crop content and acetate level in duodenum digesta were decreased by supplementation with IMO (P < 0.05) . Isovalerate level in duodenum digesta was decreased in the 0.3 and 0.6% IMO groups (P < 0.001), whereas the jejunum butyrate and isobutyrate levels of the 0.3% IMO group were higher than in other groups (P < 0.05) . The facultative microflora of the crop and cecum were not affected by IMO supplementation . However, the thymus index was increased significantly in chicks consuming diets containing 0.3% IMO. Poult Sci, 2003 Apr, 82(4), 603 - 11 Fermented liquid feed reduces susceptibility of broilers for Salmonella enteritidis; Heres L et al.; The presence of Salmonella in chickens is a problem because poultry meat is recognized as a source of human salmonellosis . Fermented feed has characteristics like a high number of lactobacilli and high concentration of lactic acid, which could make chickens less susceptible for infection with Salmonella . Fermented feed might therefore prevent the colonization of chickens with Salmonella . Two studies were performed to quantify the effect of fermented liquid feed on the susceptibility of broilers for Salmonella . The fermented feed was prepared by fermenting a dry broiler feed supplemented with 1.4 parts of water . Lactobacillus plantarum was used for fermentation . The fermented liquid feed (FLF) contained 10(9) to 10(10) cfu lactobacilli per gram, and the pH was 4 . Individually housed control chickens and FLF-fed chickens were inoculated with 10(2) to 10(7) cfu Salmonella enteritidis (SE) . Colonization was estimated by cloacal swabs and quantitative caecal culture . The proportion of SE-shedding chickens was decreased in FLF-fed chickens . FLF-fed chickens required a longer time after inoculation or a higher inoculation dose to get the same proportion of infected chickens in comparison with dry feed-fed chickens . The level of cecal colonization with Salmonella in the ceca was not different at the end of the experimental period . The results indicate that FLF can hamper the introduction of Salmonella in broiler flocks because the chickens are less susceptible for infection . Fermented liquid feed might therefore be a new hurdle in the strategy to control Salmonella in chicken flocks. Int J Gynaecol Obstet, 2003 May, 81(2), 175 - 82 Cervical Lactobacillus and leukocyte infiltration in preterm premature rupture of membranes; Silva MG et al.; OBJECTIVES: To evaluate the possible association between microorganisms present in the cervical secretions and amniotic fluid of pregnant women with preterm premature rupture of membranes (PPROM), and histologic chorioamnionitis . METHODS: Thirty-seven pregnant women with PPROM and 21 healthy pregnant women were studied . Secretions from the cervical canal and amniotic fluid were collected to isolate microorganisms present in the genital tract . Cervical smears were Gram stained and evaluated microscopically . At delivery, chorioamniotic membranes were collected for histopathologic analysis . RESULTS: Microscopic examination of the cervical secretion smears obtained from the PPROM group showed a low rate of Lactobacillus species, large numbers of leukocytes, and a wide diversity of microorganisms compared with the control group . The PPROM group presented an 80% rate of chorioamnionitis . Staphylococcus aureus isolation in cervical secretion was associated with intense inflammatory infiltrate in the membranes and might play a role in the pathogenesis of PPROM . CONCLUSIONS: The low colonization of cervical flora by Lactobacillus species associated with an intense leukocyte infiltrate detected in Gram-stained cervical smears can be considered a rapid method of detecting chorioamnionitis in pregnant women with PPROM. J Dairy Sci, 2003 Mar, 86(3), 719 - 27 Effect of Lactobacillus helveticus and Propionibacterium freudenrichii ssp . shermanii combinations on propensity for split defect in Swiss cheese; White SR et al.; One of the least controlled defects in Swiss cheese is development of splits that appear during refrigerated storage after cheese is removed from the warm room . Such fissures, or cracks, in the body of the cheese can be as short as 1 cm, or long enough to span a 90-kg block . A 2 x 2 x 2 factorial experiment was used to determine the effect of different Lactobacillus helveticus/Propionibacterium freudenreichii ssp . shermanii starter culture combinations on the occurrence of split defect in Swiss cheese . Eights vats of cheese were made in summer and eight in winter . Each 90-kg block of cheese was cut into twenty-four 4-kg blocks and graded based on the presence of splits . Only small variations were found in the composition of cheeses made during the same season . There were no correlations between moisture, pH, fat, protein, calcium, lactose contents, D/L lactate ratio, or protein degradation that could be used to predict splits after 90 d of storage . However, cheese made in the summer had 2% higher moisture content and a greater prevalence of splits . There was a sixfold increase in amount of downgraded cheese between the best and worst culture combinations used during cheese manufacture . After 90-d storage, 14 to 90% of cheese had splits in the summer, and 1 to 6% in the winter . Split formation increased with time from 60 to 120 d of storage and extent of split formation was influenced by both the lactobacilli and propionibacteria cultures used. Biotechnol Bioeng, 2003 Jun 30, 82(7), 864 - 71 Production of mannitol and lactic acid by fermentation with Lactobacillus intermedius NRRL B-3693; Saha BC et al.; Lactobacillus intermedius B-3693 was selected as a good producer of mannitol from fructose after screening 72 bacterial strains . The bacterium produced mannitol, lactic acid, and acetic acid from fructose in pH-controlled batch fermentation . Typical yields of mannitol, lactic acid, and acetic acid from 250 g/L fructose were 0.70, 0.16, and 0.12 g, respectively per g of fructose . The fermentation time was greatly dependent on fructose concentration but the product yields were not dependent on fructose level . Fed-batch fermentation decreased the time of maximum mannitol production from fructose (300 g/L) from 136 to 92 h . One-third of fructose could be replaced with glucose, maltose, galactose, mannose, raffinose, or starch with glucoamylase (simultaneous saccharification and fermentation), and two-thirds of fructose could be replaced with sucrose . L . intermedius B-3693 did not co-utilize lactose, cellobiose, glycerol, or xylose with fructose . It produced lactic acid and ethanol but no acetic acid from glucose . The bacterium produced 21.3 +/- 0.6 g lactic acid, 10.5 +/- 0.3 g acetic acid, and 4.7 +/- 0.0 g ethanol per L of fermentation broth from dilute acid (15% solids, 0.5% H(2)SO(4), 121 degrees C, 1 h) pretreated enzyme (cellulase, beta-glucosidase) saccharified corn fiber hydrolyzate . J Agric Food Chem, 2003 Apr 23, 51(9), 2745 - 52 Fluorescence labeling of wheat proteins for determination of gluten hydrolysis and depolymerization during dough processing and sourdough fermentation; Thiele C et al.; This study was undertaken to enable the determination of hydrolysis and functionality of proteins in situ during fermentation of wheat doughs . Wheat proteins were fractionated and labeled with fluorescein-isothiocyanate (FITC) . Fluorescent proteins were incorporated into wheat sourdoughs inoculated with lactobacilli and into neutral and acid control doughs . Doughs containing fungal protease were furthermore evaluated . Doughs were analyzed by extraction and size exclusion chromatography analysis of sodium dodecyl sulfate soluble proteins . Labeled proteins exhibited characteristics comparable to native proteins, with respect to proteolytic degradation and polymerization . Proteolytic breakdown of proteins was enhanced at low pH . Glutenin subunits were incorporated into the gluten macropolymer at neutral pH . Polymerization of FITC proteins was not observed at low pH . Sourdoughs were comparable to acid control doughs, major effects were attributed to changes of pH, rather than microbial metabolism . A synergistic effect with respect to proteolytic activity was observed between fungal protease and L . pontis. J Appl Microbiol, 2003, 94(5), 800 - 7 Accumulation of 1,2-propanediol and enhancement of aerobic stability in whole crop maize silage inoculated with Lactobacillus buchneri; Nishino N et al.; Aims: To assess the effects of inoculation of Lactobacillus buchneri on the ensiling properties and aerobic stability of maize silage . Methods and Results: Chopped whole crop maize was ensiled in 0.5 litre airtight polyethylene bottles (0.4 kg per bottle) and in double-layered, thin polyethylene bags (15 kg per bag), with or without inoculation of Lact . buchneri . The silos were stored for two to four months and the chemical composition, microbial numbers and aerobic stability were determined . Inoculation lowered lactic acid and yeasts, and increased acetic acid and pH value, resulting in improved aerobic stability of the silages . Inoculated silages produced 1,2-propanediol, the content of which increased as ensiling was prolonged, and nearly 50 g kg-1 dry matter had accumulated after four months of storage . The effects of inoculation, however, were much less pronounced in silages prepared in bags . Mannitol was found in all silages; the production was lowered by Lact . buchneri treatment and appeared to be unrelated to the accumulation of 1,2-propanediol . Conclusions: Inoculation of Lact . buchneri occasionally causes accumulation of 1,2-propanediol in silages without further degradation into propionic acid and 1-propanol . Significance and Impact of the Study: Substantial amounts of 1,2-propanediol could be consumed by ruminants when fed on silages inoculated with Lact . buchneri . In addition to increasing acetic acid, attention needs to be paid to 1,2-propanediol because the two fermentation products might affect the intake and utilization of silage-based diets. J Clin Periodontol, 2003 Apr, 30(4), 307 - 14 Differences in antimicrobial activity of four commercial 0.12% chlorhexidine mouthrinse formulations: an in vitro contact test and salivary bacterial counts study; Herrera D et al.; AIM: To evaluate the in vitro and in vivo antimicrobial activity of four commercial 0.12% chlorhexidine mouthrinses . MATERIAL AND METHODS: The in vitro antimicrobial activity test consisted in a modified contact test where 20 selected bacterial species were tested during 1 min with each test product . After the contact, the inoculum was cultured, and the results were expressed in terms of survival/resistance and the percentage of survival as compared to a saline control . The in vivo test consisted of a double-blind, randomized, crossover salivary bacterial counts study . 10 volunteers rinsed during 1 min with each tested product . Saliva samples were obtained before rinsing, and after 5 min, and 1, 3, 5 and 7 h . These samples were cultured both aerobically and anaerobically . Percentages of survival, in regard to baseline, were calculated for each time point . Comparisons among products were tested using anova and selected paired t-test . RESULTS: The in vitro contact test showed no survival in any tested species with CHX+CPC, while three species (Lactobacillus casei, Streptococcus mitis and Peptostreptococcus micros) were resistant to the other three products . CHX and CHX+NaF demonstrated additional resistant species (three and four species, respectively) . The in vivo salivary bacterial counts test showed higher reductions of CHX+CPC and CHX+ALC in aerobic and anaerobic bacteria, lasting for 5 h . Significant differences were detected at multiple time points, when these two products were compared both with the control and the other tested products . CONCLUSION: Important differences in activity, among 0.12% CHX products, were detected by both in vitro and in vivo tests . The formulation with alcohol was more active than those without alcohol, excepting the formulation with CHX+CPC, in which the reformulation and addition of CPC not only compensate but rather increase the antimicrobial activity. Biochemistry, 2003 Apr 22, 42(15), 4578 - 84 Epimerization at carbon-5' of (5'R)-{5'-2H}adenosylcobalamin by ribonucleoside triphosphate reductase: cysteine 408-independent cleavage of the Co-C5' bond; Chen D et al.; The adenosylcobalamin-dependent ribonucleoside triphosphate reductase (RTPR) from Lactobacillus leichmannii catalyzes the reduction of ribonucleoside triphosphates to deoxyribonucleoside triphosphates . RTPR also catalyzes the exchange of the C5'-hydrogens of adenosylcobalalamin with solvent hydrogen . A thiyl radical located on Cys 408 is generated by reaction of adenosylcobalamin at the active site and is proposed to be the intermediate for both the nucleotide reduction and the 5'-hydrogen exchange reactions . In the present research, a stereochemical approach is used to study the mechanism of the Co-C5' bond cleavage of adenosylcobalamin in the reaction of RTPR . When stereoselectively deuterated coenzyme, (5'R)-{5'-(2)H(1)} adenosylcobalamin (5'R/S = 3:1), was incubated with RTPR or the Cys 408 viariants, C408A-RTPR and C408S-RTPR in the presence of dGTP, the deuterium at the 5'-carbon was stereochemically scrambled, leading to epimerization of the (5'S)-{5'-(2)H(1)}- and (5'R)-{5'-(2)H(1)}-isotopomers . Observation of epimerization with mutated RTPR proves that transient cleavage of the Co-C5' bond occurs in the absence of the thiol group on Cys 408 . The rate constants for epimerization by RTPR, C408A-RTPR, and C408S-RTPRs in the presence of dGTP are 5.1, 0.28, and 0.42 s(-1), respectively . Only the wild-type RTPR catalyzes the 5'-hydrogen exchange reaction . Both epimerization and 5'-hydrogen exchange reactions are stimulated by the allosteric effector dGTP, and epimerization is not detected in the absence of the effector . Mechanistic implications with respect to wt-RTPR-mediated carbon cobalt bond homolysis and the intermediacy of the 5'-deoxyadenosyl radical will be presented. J Calif Dent Assoc, 2003 Mar, 31(3), 229 - 45 Fluoride-releasing restorative materials and secondary caries; Hicks J et al.; Secondary caries is responsible for 60 percent of all replacement restorations in the typical dental practice . Risk factors for secondary caries are similar to those for primary caries development . Unfortunately, it is not possible to accurately predict which patients are at risk for restoration failure . During the past several decades, fluoride-releasing dental materials have become a part of the dentist's armamentarium . Considerable fluoride is released during the setting reaction and for periods up to eight years following restoration placement . This released fluoride is readily taken up by the cavosurface tooth structure, as well as the enamel and root surfaces adjacent to the restoration . Resistance against caries along the cavosurface and the adjacent smooth surface has been shown in both in vitro and in vivo studies . Fluoride-releasing dental materials provide for improved resistance against primary and secondary caries in coronal and root surfaces . Plaque and salivary fluoride levels are elevated to a level that facilitates remineralization . In addition, the fluoride released to dental plaque adversely affects the growth of lactobacilli and mutans streptococci by interference with bacterial enzyme systems . Fluoride recharging of these dental materials is readily achieved with fluoridated toothpastes, fluoride mouthrinses, and other sources of topical fluoride . This allows fluoride-releasing dental materials to act as intraoral fluoride reservoirs . The improvement in the properties of dental materials with the ability to release fluoride has improved dramatically in the past decade, and it is anticipated that in the near future the vast majority of restorative procedures will employ fluoride-releasing dental materials as bonding agents, cavity liners, luting agents, adhesives for orthodontic brackets, and definitive restoratives. Am J Dent, 2002 Dec, 15(6), 356 - 60 Comparison of antibacterial activity of simplified adhesive systems; Imazato S et al.; PURPOSE: To determine and compare the intrinsic antibacterial activity of commercially available simplified adhesive systems . MATERIALS AND METHODS: The antibacterial activity of five self-etching/priming one-step adhesives and three priming/bonding adhesives against Streptococcus mutans, Lactobacillus casei, and Actinomyces viscosus was assessed by the agar disc-diffusion test and determination of the minimum inhibitory/bactericidal concentration (MIC/MBC) . Twenty microl of each adhesive was impregnated into a paper disc and placed on an agar plate inoculated with a bacterial suspension, with or without light-irradiation . The size of inhibition zones produced around the specimens was measured after 48 hours of incubation . The MIC values were measured by serial microdilution assays, visually examining the growth of bacteria after incubation with each adhesive for 24-48 hours . The subcultures were made on BHI agar plates from the wells showing no visible growth of bacteria, and the MBC values were determined based on production of colonies . Two primers in two-step self-etching systems and an experimental primer containing chlorhexidine were tested for reference . The results of disc-diffusion tests were analyzed by ANOVA and Fisher's PLSD test . RESULTS: The size of inhibition zones produced by adhesives varied among the brands . None of the available commercial adhesives showed significant inhibition against all three of the bacterial species tested . Light-irradiation reduced the size of inhibition zones for a few materials, but a similar trend to the effectiveness of non-irradiated specimens was observed . The antibacterial activity of commercial products determined in terms of MIC/MBC values were different from the results by disc-diffusion tests . The self-etching adhesives with low pH were not necessarily more effective than priming/bonding solutions, and no significant relationships between the acidity and antibacterial effects were found . Compared with commercial products, an experimental primer containing chlorhexidine produced significantly larger inhibition zones against all species (P < 0.05) and exhibited greater bacteriostatic/bactericidal activity, demonstrating lower MIC/MBC values. Cent Eur J Public Health, 2003 Mar, 11(1), 23 - 4 Cytolytic vaginosis: examination of 2947 vaginal smears; Demirezen S; The aim of this study is to detect the rate of cytolytic vaginosis (CV) cases in patients with symptoms resembling those ones of candida vaginitis and to distinguish them from candidiasis cases by examining of 2947 Papanicolaou-stained vaginal smears . Fifty four of 2947 patients (1.83%) were diagnosed as having CV based on cytologic criteria such as naked nuclei of intermediate cells (IC), the overgrowth of lactobacilli, cytoplasmic fragmentations due to lysis of the cells . None of these smears contained candidal blastospores and hyphae and polymorphonuclear leucocytes . The pH was 3.5 to 5.5 and the clinical symptoms were profuse vaginal discharge (VD) especially whitish-cheesy vaginal discharge and other symptoms of vaginal candidiasis . The study indicated that special attention has to be paid during evaluation the vaginal smears of the patients with presumed vaginal candidiasis in order to prevent an erroneous diagnosis of CV. J Gen Appl Microbiol, 2003 Feb, 49(1), 51 - 8 The effect of sodium acetate on the activity of L- and D-lactate dehydrogenases in Lactobacillus sakei NRIC 1071(T) and other lactic acid bacteria; Iino T et al.; The effect of sodium acetate on the production of stereoisomers of lactic acid produced by Lactobacillus sakei NRIC 1071(T) and other lactic acid bacteria was studied . L . sakei NRIC 1071(T) started producing L-lactic acid at the early logarithmic phase and d-lactic acid at the late logarithmic phase . The activity of L-lactate dehydrogenase {EC 1.1.1.27, L-LDH} from the resting cells of L . sakei NRIC 1071(T) appeared at the early stage of the logarithmic phase during the growth, and the activity of D-lactate dehydrogenase {EC 1.1.1.28, D-LDH} at the late stage of the logarithmic phase . The resting cells and cell-free extracts of L . sakei NRIC 1071(T) did not produce DL-lactic acid from L- or D-lactic acid . Stained bands of L-LDH and D-LDH appeared in the cell-free extracts from the cells of L . sakei NRIC 1071(T) . Consequently, L . sakei conclusively produced L- and D-lactic acid by the action of L-LDH and D-LDH . This finding leads to the conclusion that lactate racemase {EC 5.1.2.1} does not exist in this strain . When the specific activity of LDHs (the total activity of L-LDH plus D-LDH) from the cells cultivated in the presence of sodium acetate is compared with that cultivated in its absence, the ratio of the activity between the cells cultivated in the former condition and those in the latter fell from 1.7 on the cell-free extracts to 1.3 on the preparation of the QAE-Toyopearl 550c chromatography . This result indicates that the amount of LDHs in the cells of L . sake NRIC 1071(T) cultivated in the presence of 50 mM sodium acetate was much more than that in the cells cultivated in the absence of sodium acetate . The shift of the type of stereoisomers of lactic acid from the DL-type to the L-type is discussed in the case of L . sakei strains. J Gen Appl Microbiol, 2003 Feb, 49(1), 21 - 7 Seed culture and its effect on the growth and lactic acid production of Lactobacillus helveticus; Amrane A; Effects of seed culture medium on the subsequent culture of L . helveticus growing on whey supplemented with yeast extract and peptones have been examined . Cells were inoculated when the maximum cellular activity was achieved, i.e . the maximum for the target function, the product of the specific growth rate with the undissociated lactic acid concentration . This function decreased when the nitrogen supplementation of the preculture medium was lowered, resulting only in an increase of the lag phase length of the culture, corresponding to a cellular adaptation to the new medium . On the contrary, when cells were not in the same physiological state, growth and production parameters were affected, maximum rates, as well as maximum biomass concentration . This occurred in case of the comparison of inocula carried out under pH control or not, or when various preculture lengths were tested. Berl Munch Tierarztl Wochenschr, 2003 Mar-Apr, 116(3-4), 96 - 101 Lectin-like binding of lactobacilli considered for their use in probiotical preparations for animal use; Styriak I et al.; Three gut lactobacilli from piglets (Lactobacillus plantarum L 5, Lactobacillus paracasei L 81, Lactobacillus fermentum L 670) and Lactobacillus casei subsp . pseudoplantarum L.c.) from a calf were examined by microtitre plate binding assay for their lectin-like binding activity after their cultivation on Rogosa agar and in MRS broth . Three ECM (extracellular matrix) molecules (fetuin, porcine fibronectin and porcine mucin) were selected for this assay . Additionally, the effect of heparin on the binding of these three ECM molecules by Lactobacillus strains in microtitre plates was tested . Moreover, haemagglutination tests with pig, cattle, sheep, and hen erythrocytes were performed . However, none of the four Lactobacillus strains examined did react with any of the erythrocytes tested . The differences between individual strains were observed in their binding to immobilised ECM molecules . The best adherent was the Lactobacillus plantarum L5, however, the other three strains showed also good ECM binding . With regard to an influence of cultivation medium on lectin-like binding activity, binding of all ECM molecules was expressed in Lactobacillus paracasei L 81 to significantly higher degree (P < 0.001) after cultivation on Rogosa agar than in MRS broth . Similarly, strains Lactobacillus fermentum L 670 and Lactobacillus casei subsp . pseudoplantarum L.c . displayed significantly higher (P < 0.001) binding of fibronectin and mucin after growth on Rogosa agar in comparison with MRS broth cultivation . However, no significant (on fetuin and fibronectin binding) or opposite effect (on mucin binding) of cultivation medium was observed in Lactobacillus plantarum L 5 strain . The influence of cultivation medium on fetuin binding by Lactobacillus fermentum L 670 was also not significant while Lactobacillus casei subsp . pseudoplantarum L.c . bound fetuin significantly better (P < 0.01) after growth on Rogosa agar . Heparin pretreatment increased the binding of the ECM molecules by the Lactobacillus fermentum L 670 strain significantly (P < 0.001 or P < 0.05) with the exception of porcine fibronectin when the strain was cultivated in MRS broth . This result is important especially in the connection with the previous observations that heparin decreased ECM binding of enteropathogens as staphylococci or clinical enterococcal isolates . Following up on some earlier strain characteristics, these results indicate that the selected lactobacilli are probably suitable for probiotic purposes. J Ind Microbiol Biotechnol, 2003 Apr, 30(4), 216 - 9 Epub 2003 Apr 02. Development of Lactobacillus plantarum LL441 and its plasmid-cured derivatives in cheese; Delgado S et al.; A wild Lactobacillus plantarum strain and two of its plasmid-cured derivatives were separately used as adjunct cultures in the manufacture of a Gouda-like traditional Spanish cheese . The wild strain, LL441, harbours seven plasmids and produces a lantibiotic-like bacteriocin . The LL441-B2 derivative has lost plasmids of 40 and 80 kb and the bacteriocin-producing capability . The LL441-B11 derivative has lost in addition a 70 kb plasmid encoding active alpha- and beta-galactosidases . All three strains could be used as adjunct cultures as none of the technological and biochemical parameters of the cheeses was affected . Both the wild-type and the two derivatives were recovered from experimental cheeses up to 30 days after manufacture at similar rates of nearly 20% . Thus, the phenotypic traits under examination were not essential for L . plantarum to grow into the cheese matrix. Microbiology, 2003 Apr, 149(Pt 4), 1073 - 82 Biphasic kinetics of growth and bacteriocin production with Lactobacillus amylovorus DCE 471 occur under stress conditions; Neysens P et al.; Micro-organisms used during the production of fermented foods are subjected to several abiotic stresses . Microbial survival during these processes strongly depends on the ability of the cells to adapt and become more tolerant to the environmental conditions . Cultivation of Lactobacillus amylovorus DCE 471, a potential strain for use during type II sourdough fermentations, at low temperatures, unfavourable pH and high salt concentrations resulted in biphasic growth patterns . In addition, two separate bacteriocin peaks, as well as a dramatic change in cellular morphology, were observed . In general, an increase of the specific bacteriocin production occurred during the second growth phase . Finally, the observed sugar consumption profiles were affected by the applied fermentation temperature . Moreover, the highest bacteriocin activity occurred during maltose consumption at a low constant temperature of 28 degrees C and a constant pH of 5.4 . Plate counts from both growth phases revealed the existence of two colony types . Irregular colonies were found to outnumber smoother colonies during the first growth phase, while the second growth phase was characterized by a greater number of smooth colonies . Electron microscopy was used to investigate the observed morphological switch at the single-cell level . Single, rod-shaped cells changed into elongated cells that grew in chains . Colony and cell morphology changes coincided with the biphasic growth pattern. J Nutr Health Aging, 2003, 7(2), 75 - 7 Effect of fermented milk containing the probiotic Lactobacillus casei DN-114001 on winter infections in free-living elderly subjects: a randomised, controlled pilot study; Turchet P et al.; Probiotics are being increasingly studied for their ability to enhance host resistance to, and recovery from, infection . The probiotic strain Lactobacillus casei DN-114001 has previously been shown to reduce the incidence and duration of episodes of diarrhoea in children . Our controlled pilot study aimed to evaluate the effect of supplementation for 3 weeks with milk fermented with yoghurt cultures and L . casei DN-114001 on the incidence and severity of winter infections (gastrointestinal and respiratory) in elderly people . We found no difference in the incidence of winter infections between groups . However, duration of all pathologies was significantly lower in the treatment group (7.0 3.2 days, n=180) than in the control group (8.7 3.7 days; n=180) (p=0.024), as was maximal temperature (38.3 0.5 C treatment group vs . 38.5 0.6 C control; p=0.01) . The potential for a 20% reduction in the duration of winter infections that we have found warrants further investigation on a larger scale. J Biol Chem, 2003 Jun 20, 278(25), 22861 - 7 Epub 2003 Apr 04. Kinetic and mechanistic characterization of recombinant Lactobacillus viridescens FemX (UDP-N-acetylmuramoyl pentapeptide-lysine N6-alanyltransferase); Hegde SS et al.; The FemABX family encodes enzymes that incorporate l-amino acids into the interchain peptide bridge of Gram-positive cell wall peptidoglycan and are novel nonribosomal peptidyl transferases that use aminoacyl-tRNA as the amino acid donor . We previously reported the identification of the femX gene from Lactobacillus viridescens and recombinant expression of active FemX (LvFemX) that catalyzes the transfer of l-Ala from Ala-tRNAAla to the epsilon-amino group of l-lysine of UDP-MurNAc pentapeptide (Hegde, S . S., and Shrader, T . E . (2001) J . Biol . Chem . 276, 6998-7003) . Recombinant LvFemX exhibits Km values of 42 and 15 microm for UDP-MurNAc pentapeptide and Escherichia coli Ala-tRNAAla, respectively, and exhibited a kcat value of 660 min-1 . Initial velocity and inhibition kinetic studies support an ordered sequential mechanism for the enzyme, and we propose that catalysis proceeds via a ternary complex . The pH dependence of the activity was bell-shaped, depending on the ionization state of two groups exhibiting apparent pKa values of 5.5 and 9.3 . Chemical modification of the enzyme and the kinetics of inactivation, and protection by substrate, indicated the involvement of carboxyl groups in the catalytic function of the enzyme . Site-directed mutagenesis identified Asp109 as a candidate for the catalytic base and Glu320 plays an additional important role in the catalytic function of the enzyme. Appl Environ Microbiol, 2003 Apr, 69(4), 2321 - 9 Rope-producing strains of Bacillus spp . from wheat bread and strategy for their control by lactic acid bacteria; Pepe O et al.; Two types of white wheat bread (high- and low-type loaves) were investigated for rope spoilage . Thirty of the 56 breads tested developed rope spoilage within 5 days; the high-type loaves were affected by rope spoilage more than the low-type loaves . Sixty-one Bacillus strains were isolated from ropy breads and were characterized on the basis of their phenotypic and genotypic traits . All of the isolates were identified as Bacillus subtilis by biochemical tests, but molecular assays (randomly amplified polymorphic DNA PCR assay, denaturing gradient gel electrophoresis analysis, and sequencing of the V3 region of 16S ribosomal DNA) revealed greater Bacillus species variety in ropy breads . In fact, besides strains of B . subtilis, Bacillus licheniformis, Bacillus cereus, and isolates of Bacillus clausii and Bacillus firmus were also identified . All of the ropy Bacillus isolates exhibited amylase activity, whereas only 32.4% of these isolates were able to produce ropiness in bread slices after treatment at 96 degrees C for 10 min . Strains of lactic acid bacteria previously isolated from sourdough were first selected for antirope activity on bread slices and then used as starters for bread-making experiments . Prevention of growth of approximately 10(4) rope-producing B . subtilis G1 spores per cm(2) on bread slices for more than 15 days was observed when heat-treated cultures of Lactobacillus plantarum E5 and Leuconostoc mesenteroides A27 were added . Growth of B . subtilis G1 occurred after 7 days in breads started with Saccharomyces cerevisiae T22, L . plantarum E5, and L . mesenteroides A27. Appl Environ Microbiol, 2003 Apr, 69(4), 2230 - 6 Surface display of the receptor-binding region of the Lactobacillus brevis S-layer protein in Lactococcus lactis provides nonadhesive lactococci with the ability to adhere to intestinal epithelial cells; Avall-Jaaskelainen S et al.; Lactobacillus brevis is a promising lactic acid bacterium for use as a probiotic dietary adjunct and a vaccine vector . The N-terminal region of the S-layer protein (SlpA) of L . brevis ATCC 8287 was recently shown to mediate adhesion to various human cell lines in vitro . In this study, a surface display cassette was constructed on the basis of this SlpA receptor-binding domain, a proteinase spacer, and an autolysin anchor . The cassette was expressed under control of the nisA promoter in Lactococcus lactis NZ9000 . Western blot assay of lactococcal cell wall extracts with anti-SlpA antibodies confirmed that the SlpA adhesion domain of the fusion protein was expressed and located within the cell wall layer . Whole-cell enzyme-linked immunosorbent assay and immunofluorescence microscopy verified that the SlpA adhesion-mediating region was accessible on the lactococcal cell surface . In vitro adhesion assays with the human intestinal epithelial cell line Intestine 407 indicated that the recombinant lactococcal cells had gained an ability to adhere to Intestine 407 cells significantly greater than that of wild-type L . lactis NZ9000 . Serum inhibition assay further confirmed that adhesion of recombinant lactococci to Intestine 407 cells was indeed mediated by the N terminus-encoding part of the slpA gene . The ability of the receptor-binding region of SlpA to adhere to fibronectin was also confirmed with this lactococcal surface display system . These results show that, with the aid of the receptor-binding region of the L . brevis SlpA protein, the ability to adhere to gut epithelial cells can indeed be transferred to another, nonadhesive, lactic acid bacterium. Appl Environ Microbiol, 2003 Apr, 69(4), 2217 - 22 Metabolism of fructooligosaccharides by Lactobacillus paracasei 1195; Kaplan H et al.; Fermentation of fructooligosaccharides (FOS) and other oligosaccharides has been suggested to be an important property for the selection of bacterial strains used as probiotics . However, little information is available on FOS transport and metabolism by lactic acid bacteria and other probiotic bacteria . The objectives of this research were to identify and characterize the FOS transport system of Lactobacillus paracasei 1195 . Radiolabeled FOS was synthesized enzymatically from {(3)H}sucrose and purified by column and thin-layer chromatography, yielding three main products: glucose (G) alpha-1,2 linked to two, three, or four fructose (F) units (GF(2), GF(3), and GF(4), respectively) . FOS hydrolysis activity was detected only in cell extracts prepared from FOS- or sucrose-grown cells and was absent in cell supernatants, indicating that transport must precede hydrolysis . FOS transport assays revealed that the uptake of GF(2) and GF(3) was rapid, whereas little GF(4) uptake occurred . Competition experiments showed that glucose, fructose, and sucrose reduced FOS uptake but that other mono-, di-, and trisaccharides were less inhibitory . When cells were treated with sodium fluoride, iodoacetic acid, or other metabolic inhibitors, FOS transport rates were reduced by up to 60%; however, ionophores that abolished the proton motive force only slightly decreased FOS transport . In contrast, uptake was inhibited by ortho-vanadate, an inhibitor of ATP-binding cassette transport systems . De-energized cells had low intracellular ATP concentrations and had a reduced capacity to accumulate FOS . These results suggest that FOS transport in L . paracasei 1195 is mediated by an ATP-dependent transport system having specificity for a narrow range of substrates. Appl Environ Microbiol, 2003 Apr, 69(4), 2087 - 93 Identification of equine cecal bacteria producing amines in an in vitro model of carbohydrate overload; Bailey SR et al.; Acute laminitis has been associated with the overgrowth of gram-positive bacteria within the equine hindgut, causing the release of factor(s) leading to ischemia-reperfusion of the digits . The products of fermentation which trigger acute laminitis are, as yet, unknown; however, vasoactive amines are possible candidates . The objectives of this study were to use an in vitro model of carbohydrate overload to study the change in populations of cecal streptococci and lactobacilli and to establish whether certain species of these bacteria were capable of producing vasoactive amines from amino acids . Cecal contents from 10 horses were divided into aliquots and incubated anaerobically with either corn starch or inulin (fructan; both at 1 g/100 ml) . Samples were taken at 6-h intervals over a 24-h period for enumeration of streptococci, lactobacilli, and gram-negative anaerobes by a dilution method onto standard selective growth media . The effects of the antibiotic virginiamycin (1 mg/100 ml) and calcium hydrogen phosphate (CaHPO(4); 0.3 g/100 ml) were also examined . Fermentation of excess carbohydrate was associated with increases in numbers of streptococci and lactobacilli (2- to 3.5-log unit increases; inhibited by virginiamycin) but numbers of gram-negative anaerobes were not significantly affected . A screening agar technique followed by 16S rRNA gene sequence analysis enabled the identification of 26 different bacterial strains capable of producing one or more vasoactive amines . These included members of the species Streptococcus bovis and five different Lactobacillus spp . These data suggest that certain bacteria, whose overgrowth is associated with carbohydrate fermentation, are capable of producing vasoactive amines which may play a role in the pathogenesis of acute laminitis. Appl Environ Microbiol, 2003 Apr, 69(4), 2073 - 9 Exopolysaccharide and kestose production by Lactobacillus sanfranciscensis LTH2590; Korakli M et al.; The effect was investigated of sucrose concentration on sucrose metabolism and on the formation of exopolysaccharide (EPS) by Lactobacillus sanfranciscensis LTH2590 in pH-controlled fermentations with sucrose concentrations ranging from 20 to 160 g liter(-1) . The EPS production increased and the relative sucrose hydrolysis activity decreased by increasing the sucrose concentration in the medium . The carbon recovery decreased from 95% at a sucrose concentration of 30 g liter(-1) to 58% at a sucrose concentration of 160 g liter(-1) because of the production of an unknown metabolite by L . sanfranciscensis . This metabolite was characterized as a fructo-oligosaccharide . The oligosaccharide produced by L . sanfranciscensis was purified and characterized as a trisaccharide with a glucose/fructose ratio of 1:2 . The comparison of the retention time of this oligosaccharide and that of pure oligosaccharide standards using two different chromatography methods revealed that the oligosaccharide produced by L . sanfranciscensis LTH2590 is 1-kestose . Kestose production increased concomitantly with the initial sucrose concentration in the medium. Appl Environ Microbiol, 2003 Apr, 69(4), 2044 - 51 Identification of Lactobacillus reuteri genes specifically induced in the mouse gastrointestinal tract; Walter J et al.; Lactobacilli are common inhabitants of the gastrointestinal tracts of mammals and have received considerable attention due to their putative health-promoting properties . Little is known about the traits that enhance the ability of these bacteria to inhabit the gastrointestinal tract . In this paper we describe the development and application of a strategy based on in vivo expression technology (IVET) that enables detection of Lactobacillus reuteri genes specifically induced in the murine gut . A plasmid-based system was constructed containing 'ermGT (which confers lincomycin resistance) as the primary reporter gene for selection of promoters active in the gastrointestinal tract of mice treated with lincomycin . A second reporter gene, 'bglM (beta-glucanase), allowed differentiation between constitutive and in vivo inducible promoters . The system was successfully tested in vitro and in vivo by using a constitutive promoter . Application of the IVET system with chromosomal DNA of L . reuteri 100-23 and reconstituted lactobacillus-free mice revealed three genes induced specifically during colonization . Two of the sequences showed homology to genes encoding xylose isomerase (xylA) and peptide methionine sulfoxide reductase (msrB), which are involved in nutrient acquisition and stress responses, respectively . The third locus showed homology to the gene encoding a protein whose function is not known . Our IVET system has the potential to identify genes of lactobacilli that have not previously been functionally characterized but which may be essential for growth of these bacteria in the gastrointestinal ecosystem. Cell Microbiol, 2003 Apr, 5(4), 277 - 85 Lactobacillus rhamnosus GG decreases TNF-alpha production in lipopolysaccharide-activated murine macrophages by a contact-independent mechanism; Pena JA et al.; Animal studies and human clinical trials have shown that Lactobacillus can prevent or ameliorate inflammation in chronic colitis . However, molecular mechanisms for this effect have not been clearly elucidated . We hypothesize that lactobacilli are capable of downregulating pro-inflammatory cytokine responses induced by the enteric microbiota . We investigated whether lactobacilli diminish production of tumour necrosis factor alpha (TNF-alpha) by the murine macrophage line, RAW 264.7 gamma (NO-), and alter the TNF-alpha/interleukin-10 (IL-10) balance, in vitro . When media conditioned by Lactobacillus rhamnosus GG (LGG) are co-incubated with lipopolysaccharide (LPS) or lipoteichoic acid (LTA), TNF-alpha production is significantly inhibited compared to controls, whereas IL-10 synthesis is unaffected . Interestingly, LGG-conditioned media also decreases TNF-alpha production of Helicobacter-conditioned media-activated peritoneal macrophages . Lactobacillus species may be capable of producing soluble molecules that inhibit TNF-alpha production in activated macrophages . As overproduction of pro-inflammatory cytokines, especially TNF-alpha, is implicated in pathogenesis of chronic intestinal inflammation, enteric Lactobacillus-mediated inhibition of pro-inflammatory cytokine production and alteration of cytokine profiles may highlight an important immunomodulatory role for commensal bacteria in the gastrointestinal tract. Pol J Vet Sci, 2003, 6(1), 29 - 39 Antagonistic effect of chosen lactic acid bacteria strains on Salmonella species in meat and fermented sausages; Gomolka-Pawlicka M et al.; The aim of this study was to determine of influence of 15 strains of lactic acid bacteria on the growth of 7 Salmonella spp . strains in model set-ups, and in meat and ripened fermented sausages . The investigations were performed within the framework of three alternate stages which differed in respect to the products studied, the number of Lactobacillus spp . strains and, partly, methodological approach . The ratio between lactic acid bacteria and Salmonella strains studied was, depending on the alternate, 1:1, 1:2 and 2:1, respectively . The investigations also covered the water activity (a(w)) and pH of the tested products . The results obtained are shown in 12 figures and suggest that all the lactic acid bacteria strains used within the framework of the model set-ups showed antagonistic effect on all the Salmonella spp . strains . However, these abilities were not observed with respect to some lactic acid bacteria strains in meat and fermented sausage . The temperature and length of the incubation period of sausages, but not a(w) and pH, were found to have a distinct influence on the antagonistic interaction between the bacteria. Can J Microbiol, 2003 Jan, 49(1), 51 - 7 Lactobacillus growth and membrane composition in the presence of linoleic or conjugated linoleic acid; Jenkins JK et al.; Five Lactobacillus strains of intestinal and food origins were grown in MRS broth or milk containing various concentrations of linoleic acid or conjugated linoleic acid (CLA) . The fatty acids had bacteriostatic, bacteriocidal, or no effect depending on bacterial strain, fatty acid concentration, fatty acid type, and growth medium . Both fatty acids displayed dose-dependent inhibition . All strains were inhibited to a greater extent by the fatty acids in broth than in milk . The CLA isomer mixture was less inhibitory than linoleic acid . Lactobacillus reuteri ATCC 55739, a strain capable of isomerizing linoleic acid to CLA, was the most inhibited strain by the presence of linoleic acid in broth or milk . In contrast, a member of the same species, L . reuteri ATCC 23272, was the least inhibited strain by linoleic acid and CLA . All strains increased membrane linoleic acid or CLA levels when grown with exogenous fatty acid . Lactobacillus reuteri ATCC 55739 had substantial CLA in the membrane when the growth medium was supplemented with linoleic acid . No association between level of fatty acid incorporation into the membrane and inhibition by that fatty acid was observed. Biotechnol Bioeng, 2003 Jun 20, 82(6), 653 - 63 Metabolic engineering of Lactobacillus fermentum for production of mannitol and pure L-lactic acid or pyruvate; Aarnikunnas J et al.; For production of mannitol in combination with pure L-lactic acid or pyruvate, the D- and L-lactate dehydrogenase genes (ldhD and ldhL) of a mannitol-producing Lactobacillus fermentum strain were cloned and stepwise inactivated . For inactivation of both ldh genes by a gene replacement technique, deletion constructs removing a 0.4-kb fragment from the promoter and the 5' end region of the ldh genes were used . The first inactivation mutant, designated L . fermentum GRL1030, carried the deletion in ldhD (DeltaldhD) . A double mutant, DeltaldhD-DeltaldhL, was constructed by the inactivation of the ldhL gene of strain GRL1030, resulting in strain L . fermentum GRL1032 . The correctness of the both mutants was confirmed at the DNA level by polymerase chain reaction, as shown by the absence of ldh transcripts by northern blotting and as a lack of the corresponding enzyme activity . In bioreactor cultivations, the single mutant GRL1030 produced mannitol and L-lactic acid as expected . Mannitol and lactic acid yields and productivities were practically unaffected by deletion of the ldhD gene . The double mutant GRL1032 produced mannitol and pyruvate as expected . However, although the yield of mannitol from fructose remained high, its volumetric productivity was reduced . The double mutation negatively affected the glucose consumption rate, resulting in reduced cellular growth . In addition to pyruvate, the double mutant produced 2,3-butanediol . More surprisingly, some lactic acid was still produced . FEMS Microbiol Lett, 2003 Mar 28, 220(2), 281 - 6 Purification and characterisation of mannitol dehydrogenase from Lactobacillus sanfranciscensis; Korakli M et al.; Mannitol dehydrogenase (MDH) was purified and characterised from Lactobacillus sanfranciscensis . Two peptide fragments of MDH were N-terminally sequenced for the first time in the genus Lactobacillus . The purified enzyme had an apparent molecular mass of 44 kDa and catalysed both the reduction of fructose to mannitol and the oxidation of mannitol to fructose . The K(m) value for the reduction reaction was 24 mM fructose and that for the oxidation 78 mM mannitol . The optimum temperature was 35 degrees C, the pH optima for the reduction or oxidation were 5.8 and 8, respectively. J Chromatogr B Analyt Technol Biomed Life Sci, 2003 May 5, 788(1), 187 - 91 Measurements of sub-nanomolar concentrations of unmetabolised folic acid in serum; Sweeney MR et al.; We describe a combined HPLC/microbiological assay procedure for the sub-nanomolar analysis of unmetabolised folic acid (pteroylglutamate) in human serum . This metabolically unaltered form of the vitamin arises following the consumption of folic acid either in supplemental form or in fortified foods . Following HPLC separation of folic acid from other folate derivatives the folic acid fraction was concentrated by C(18) Sep-Pak cartridges and assayed by Lactobacillus casei microbiological assay . The present assay allows the quantitation and kinetic analysis of the effects of consumption of folic acid. Acta Otorrinolaringol Esp, 2002 Dec, 53(10), 791 - 3 {Submental abscess by Lactobacillus acidophilus/jensenii}; Fajardo M et al.; The different species of Lactobacillus genus are bacteria of a low pathogenicity . They are found in the gastrointestinal tract, the female genitourinary tract, and also as part of the anaerobic flora in the mouth . Due to this low pathogenicity, they are generally related with infections together with other with more virulent microorganisms, or in immunocompromised patients . In our case, Lactobacillus appears alone as an etiological agent in an immunocompetent patient with a submental abscess. J Nutr Sci Vitaminol (Tokyo), 2002 Oct, 48(5), 325 - 31 Characterization and bioavailability of vitamin B12-compounds from edible algae; Watanabe F et al.; Substantial amounts of vitamin B12 were found in some edible algae (green and purple lavers) and algal health food (chlorella and spirulina tablets) using the Lactobacillus delbrueckii subsp . lactis ATCC7830 microbiological assay method . Corrinoid-compounds were purified and characterized from these algae to clarify the chemical properties and bioavailability of the algal vitamin B12 . True vitamin B12 is the predominate cobamide of green and purple lavers and chlorella tablets . Feeding the purple laver to vitamin B12-deficient rats significantly improved the vitamin B12 status . The results suggest that algal vitamin B12 is a bioavailable source for mammals . Pseudovitamin B12 (an inactive corrinoid) predominated in the spirulina tablets, which are not suitable for use as a vitamin B12 source, especially for vegetarians . algal health food, bioavailability, cobalamin, edible algae, vitamin B12 Int J Syst Evol Microbiol, 2003 Jan, 53(Pt 1), 263 - 8 Lactobacillus thermotolerans sp . nov., a novel thermotolerant species isolated from chicken faeces; Niamsup P et al.; Five strains of thermotolerant lactic acid bacteria (G 12, G 22, G 35T, G 43 and G 44) isolated from chicken faeces were characterized taxonomically . The strains were facultatively anaerobic, Gram-positive, catalase-negative, non-motile, non-spore-forming rods . They were heterofermentative lactobacilli that produced DL-lactic acid . Growth of the strains occurred at 45 degrees C but not at 15 degrees C . The optimum temperature for growth was 42 degrees C, as determined from the specific growth rate . The highest permissive temperatures for growth were 50 degrees C for strain G35T and 48 degrees C for the other four strains . DNA G+C content of the strains was between 49 and 51 mol% . Complex fatty acid patterns of the strains showed the presence of C14:0, C16:0, sometimes C18:0, C18:1 and C19:0 cyclo in the cell walls . Phylogenetic analysis of the 16S rRNA gene sequences of the five strains placed them in the Lactobacillus caseil Pediococcus group, with Lactobacillus fermentum as their closest relative (about 95% sequence similarity) . DNA-DNA hybridization data indicated that the thermotolerant strains were not L . fermentum . Taken together, the findings of this study show that the five strains isolated from chicken faeces represent a novel species within the genus Lactobacillus, for which the name Lactobacillus thermotolerans is proposed (G 35T = DSM 14792T =JCM 11425T). Int J Syst Evol Microbiol, 2003 Jan, 53(Pt 1), 133 - 6 Lactobacillus ingluviei sp . nov., isolated from the intestinal tract of pigeons; Baele M et al.; Lactic acid bacteria were isolated from the crop and intestines of pigeons . One group of strains, showing similar genomic patterns after screening with tRNA intergenic spacer PCR, could not be identified to the species level . Sequencing of the 16S rRNA gene of one representative strain revealed about 96% similarity to sequences from Lactobacillus fermentum and Lactobacillus mucosae . Determination of the DNA base composition, DNA-DNA hybridization experiments, SDS-PAGE of whole-cell proteins and biochemical testing confirmed that the seven strains studied constitute a single novel Lactobacillus species, for which the name Lactobacillus ingluviei sp . nov . is proposed . The type strain is strain KR3T (=LMG 20380T =CCUG 45722T). Int J Syst Evol Microbiol, 2003 Jan, 53(Pt 1), 67 - 75 Significant differences between Lactobacillus casei subsp . casei ATCC 393T and a commonly used plasmid-cured derivative revealed by a polyphasic study; Acedo-Felix E et al.; Many studies on Lactobacillus casei subsp . casei (L . casei) have been carried out using strain ATCC 393 (pLZ15-) . Four strains of L . casei ATCC 393T and three of ATCC 393 (pLZ15-) were compared using phenotypic methods and many of the available genotyping techniques . These tests showed that strains of ATCC 393T obtained from independent public type-culture collections were significantly different from the plasmid-free (pLZ15-) strains of ATCC 393T . These findings were confirmed by sequencing the first 580 nt (domain I) of the 16S and 23S rDNAs of the strains . Complete sequencing of the 16S rDNA of one representative strain from each group revealed that strain ATCC 393T from culture collections was 99% similar to Lactobacillus zeae ATCC 15820T and that the strain so far considered as L . casei ATCC 393 (pLZ15-) was, in turn, 100% similar to L . casei ATCC 334 and Lactobacillus paracasei subsp . paracasei ATCC 4022 . All data obtained in this work indicate that the ancestral strain of ATCC 393 (pLZ15-) might never have been the strain that is now available from culture collections. Int J Syst Evol Microbiol, 2003 Jan, 53(Pt 1), 7 - 13 Molecular analysis of sourdough reveals Lactobacillus mindensis sp . nov; Ehrmann MA et al.; Genotypic fingerprinting to analyse the bacterial flora of an industrial sourdough revealed a coherent group of strains which could not be associated with a valid species . Comparative 16S rDNA sequence analysis showed that these strains formed a homogeneous cluster distinct from their closest relatives, Lactobacillus farciminis, Lactobacillus alimentarius and Lactobacillus kimchii . To characterize them further, physiological (sugar fermentation, formation of DL-lactate, hydrolysis of arginine, growth temperature, CO2 production) and chemotaxonomic properties have been determined . The DNA G +C content was 37.5 0.2 mol% . The peptidoglycan was of the lysine-D-iso-asparagine (L-Lys-D-Asp) type . The strains were homofermentative, Gram-positive, catalase-negative, non-spore-forming, non-motile rods . They were found as a major stable component of a rye flour sourdough fermentation . Physiological, biochemical as well as genotypic data suggested them to be a new species of the genus Lactobacillus . This was confirmed by DNA-DNA hybridization of genomic DNA, and the name Lactobacillus mindensis is proposed . The type strain of this species is DSM 14500T (=LMG 21508T). Caries Res, 2003 Mar-Apr, 37(2), 142 - 7 Metabolic control as a modifier of the association between salivary factors and dental caries among diabetic patients; Syrjala AM et al.; The aim here was to analyze the role of metabolic control as a modifier of the association of salivary factors with dental caries among diabetic patients . Cross-sectional data were gathered from 149 insulin-dependent diabetes mellitus patients by means of clinical examination, a quantitative questionnaire and patient records . Dental caries was measured as the number of decayed surfaces . The data were analyzed using negative binomial regression models . HbA1(c) level appeared not to be associated with dental caries . Of the salivary factors, only high levels of mutans streptococci (MS) and lactobacilli were associated with dental caries . Among the subjects with HbA1(c) > or =8.5, the presence of dental caries was more distinctly associated with higher levels of MS (RR = 3.86, CI: 1.39-11.96) and lactobacilli (RR = 3.59, CI: 1.44-9.33) compared to those with HbA1(c)<8.5 . It can be concluded that poor glycemic control strengthens the positive association of MS and lactobacilli with dental caries . Caries Res, 2003 Mar-Apr, 37(2), 108 - 14 Inhibitory effects on selected oral bacteria of antibacterial agents incorporated in a glass ionomer cement; Botelho MG; The objectives of the study were to investigate the antimicrobial efficacy, over time, of combining antibacterial agents with a glass ionomer cement (GIC) . This was assessed using an agar diffusion test . Chlorhexidine hydrochloride, cetylpyridinium chloride, cetrimide and benzalkonium chloride were added to Fuji IX GIC at 0, 1, 2 and 4% w/w . Antibacterial-GIC specimens were placed onto agar plates inoculated with one of six bacterial species (Streptococcis, Lactobacillus, and Actinomyces, two each) and the area of inhibition calculated after 24 h incubation . The experiment was repeated weekly and at week 11 the surface of the specimen was abraded prior to replacing on inoculated agar plates . Control specimens of the GIC produced no bacterial inhibition . The antibacterial-GIC combination specimens showed significant inhibition which decreased at different rates over the test period . Resurfacing of the specimens showed a dramatic increase of antibacterial action similar to levels produced on week 1 . CT-GIC showed the greatest (p < 0.005) inhibitory effect throughout the experimental period for 4 out of 6 test bacteria . The addition of antibacterial agents to Fuji IX creates a GIC material with significant antimicrobial action in vitro which is dependent on concentration and type of antibacterial agent, and appears to be associated primarily with a release of the antibacterial from the surface layer of the specimen . J Dairy Sci, 2003 Feb, 86(2), 429 - 38 The impact of fermentation and in vitro digestion on the formation of angiotensin-I-converting enzyme inhibitory activity from pea and whey protein; Vermeirssen V et al.; Pea and whey protein were fermented by Lactobacillus helveticus and Saccharomyces cerevisiae in monoculture and in combination at 28 and 37 degrees C in order to release angiotensin-I-converting enzyme (ACE) inhibitory peptides . The fermentation products were subjected to in vitro gastrointestinal digestion, and the digests of nonfermented samples served as controls . After fermentation, the ACE inhibitory activity (%) increased by 18 to 30% for all treatments, except for the fermentations of whey protein with Saccharomyces cerevisiae at 28 degrees C, where no significant change was observed . After digestion, however, both fermented and nonfermented samples reached maximum ACE inhibitory activity . The whey digests tended to have lower (50%) inhibitory concentrations (IC50; 0.14 to 0.07 mg/ml), hence, higher ACE inhibitory activity, than the pea digests (0.23 to 0.11 mg/ml) . The nonfermented whey protein digest showed the highest ACE inhibitory activity of all . For pea protein, the nonfermented sample had the lowest IC50 value . These results suggest that in vitro gastrointestinal digestion was the predominant factor controlling the formation of ACE inhibitory activity, hence, indicating its importance in the bioavailability of ACE inhibitory peptides. J Dairy Sci, 2003 Feb, 86(2), 424 - 8 Increase of viability of entrapped cells of Lactobacillus delbrueckii ssp . bulgaricus in artificial sesame oil emulsions; Hou RC et al.; A technique was developed to protect lactic acid bacteria (Lactobacillus delbrueckii ssp . bulgaricus) against simulated gastrointestinal conditions by encapsulation of bacterial cells within artificial sesame oil emulsions . Purified sesame oil bodies consisting of approximately 99% oil, 0.5% phospholipid, and 0.5% protein were decomposed by heating at 70 degrees C for 1 h . The bacteria cultured in nonfat milk were encapsulated in artificial oil emulsions constituted with decomposed sesame oil bodies and excess sesame or vegetable cooking oil . Viability of bacteria in storage at 4 degrees C for 16 d was substantially elevated from 0.023 to 5.45% after encapsulation . Compared with free cells, the entrapped bacteria demonstrated a significant increase (approximately 10(4) times) in survival rate when subjected to simulated high acid gastric or bile salt conditions . The results indicate that artificial sesame oil emulsion may serve as an effective biocapsule for encapsulation of bacteria in dairy products. Carbohydr Res, 2003 Mar 28, 338(7), 605 - 9 Structural analysis of the Lactobacillus rhamnosus strain KL37C exopolysaccharide; Lipinski T et al.; The exopolysaccharide from the lactic acid bacterium Lactobacillus rhamnosus strain KL37C isolated from human intestinal flora was prepared by sonication of bacterial cell mass suspended in water followed by centrifugation and cold ethanol precipitation of the supernatant . The polysaccharide material was purified by gel permeation chromatography on an TSK HW-50 column and characterised using chemical and enzymatic methods . On the basis of sugar and methylation analysis and 1H, 13C, 1D and 2D NMR spectroscopy the exopolysaccharide was shown to be composed of the following pentasaccharide repeating unit:-->3)-alpha-D-Glcp-(1-->2)-beta-D-Galf-(1-->6)-alpha-D-Galp-(1-->6)-alpha-D-Glcp-(1-->3)-beta-D-Galf-(1--> FEMS Microbiol Lett, 2003 Mar 14, 220(1), 121 - 5 Bacteriocin-like substance (BLS) production in Aeromonas hydrophila water isolates; Messi P et al.; 30 Aeromonas hydrophila water isolates were tested for bacteriocin-like substance (BLS) production using a target panel of closely related microorganisms and other Gram-positive and Gram-negative bacteria, including food-borne pathogens . A . hydrophila showed antibacterial activity against one or more indicator microorganisms, but the activity emerged only with non-phylogenetically related genera or species . In particular all A . hydrophila showed antibacterial activity against one or more of the tested Staphylococcus strains, five against Listeria spp . (Listeria seeligeri, Listeria welshimeri and Listeria ivanovii), and eight presented a weak antagonistic activity towards Streptococcus agalactiae and Lactobacillus spp . Inhibitory activity was not observed against the other Gram-positive (Listeria monocytogenes, Listeria innocua and Enterococcus spp.) and Gram-negative tested strains, including Aeromonas sobria, Aeromonas caviae and the same A . hydrophila, when used as indicator . Anti-staphylococcal activity was observed with a gradual increase of the inhibition zone during incubation and seemed to be influenced by A . hydrophila hemolytic expression . Extrachromosomal analysis showed the presence, in 70% of the strains, of one to five plasmids with molecular masses ranging from 2.1 to 41.5 MDa, but it was not possible to relate this result with BLS production. J Calif Dent Assoc, 2003 Feb, 31(2), 135 - 8 Acquisition and transmission of mutans streptococci; Berkowitz RJ; Dental caries is an infectious and transmissible disease . The mutans streptococci and some Lactobacillus species are the two groups of infectious agents most strongly associated with dental caries . Earlier studies demonstrated that infants acquire mutans streptococci from their mothers and only after the eruption of primary teeth . More recent studies indicate that mutans streptococci can colonize the mouths of predentate infants and that horizontal, as well as vertical, transmission does occur . These findings will likely facilitate the development of clinical strategies to prevent or delay infant infection by these organisms, thereby reducing the prevalence of dental caries. J Food Prot, 2003 Mar, 66(3), 466 - 72 Interactions of lactic acid bacteria with human intestinal epithelial cells: effects on cytokine production; Wallace TD et al.; As a participant in the mucosal immune response, the intestinal epithelial cell must respond to a variety of stimuli, including lactic acid bacteria (LAB) consumed in the diet . The objective of this study was to compare the abilities of several strains of LAB to modulate cytokine secretion by human intestinal epithelial cell (IEC) line HT-29 . Certain strains of Lactobacillus rhamnosus, Lactobacillus delbrueckii, and Lactobacillus acidophilus suppressed the production of the chemokine RANTES by stimulated HT-29 IEC, although the magnitude of this suppression varied depending on the nature of the bacterial growth medium . Similarly, specific strains showed growth condition-dependent suppression of HT-29 interleukin-8 (IL-8) production . Strain-dependent effects were also seen for the suppression of tumor necrosis factor alpha (TNF-alpha) and transforming growth factor beta (TGF-beta) production . The binding of several of these bacterial strains to the HT-29 cell line was also examined . Different strains were found to have differing abilities to interact with IEC, with L . rhamnosus R0011 being the strain that generally had the most extensive effects on HT-29 cytokine production and also bound to HT-29 IEC most effectively . Modulation of IEC cytokine production has the potential to profoundly affect the mucosal microenvironment, influencing the immune response to pathogens and other ingested antigens. J Food Prot, 2003 Mar, 66(3), 426 - 30 Kinetics of adsorption and desorption of aflatoxin B1 by viable and nonviable bacteria; Lee YK et al.; The reactions involved in the binding (adsorption) and release (desorption) of aflatoxin B1 (AFB1) to and from the surface of bacteria were investigated . Viable and heat-killed Lactobacillus rhamnosus GG, L . rhamnosus LC-705, and Propionibacterium freudenreichii subsp . shermanii JS were incubated in phosphate-buffered saline containing variable concentrations (0.0017 to 13.3 microg/ml) of AFB1 . The relationship between the bacterial surface hydrophobicity and the AFB1 adsorption affinity was also investigated . A linear relationship was observed between the specific rate of AFB1 adsorption and the AFB1 concentration for all bacteria . The nature of desorption of adsorbed AFB1 was investigated by repetitive aqueous washes . A linear relationship was observed between the natural log value of the concentration of AFB1 adsorbed and the number of washes for all bacteria studied . The desorption constants were strain-dependent and were lower for heat-killed bacteria than for viable bacteria . Heat treatment appears to alter the surface properties of the bacteria rather than expose new adsorption sites . No correlation was found between the hydrophobicity and the AFB1 adsorption affinity. J Food Prot, 2003 Mar, 66(3), 418 - 25 Antagonistic action of cells of Lactobacillus delbrueckii subsp . lactis against pathogenic and spoilage microorganisms in fresh meat systems; Senne MM et al.; Cells of Lactobacillus delbrueckii subsp . lactis RM2-5 were added to various meat model systems that had been inoculated with Escherichia coli O157:H7 or Salmonella Typhimurium to determine whether these lactobacilli were antagonistic to the pathogens during storage at 5 degrees C . Experiments in which L . delbrueckii subsp . lactis RM2-5 was directly applied to the surfaces of beefsteaks resulted in significant (P < 0.05) reductions in the growth of psychrotrophs and coliforms plus a slight decrease in the numbers of E . coli O157:H7 over time relative to those for control samples to which no lactobacilli had been added . Experiments involving the direct application of L . delbrueckii subsp . lactis RM2-5 to the surfaces of freshly slaughtered beef and pork carcass samples inoculated with either E . coli O157:H7 or Salmonella Typhimurium showed significant (P < 0.05) declines in numbers of the pathogens as well as a reduction in the growth of psychrotrophs during storage at 5 degrees C for 6 days . The results of the experiments suggest that lactobacillus cultures have potential for use in an intervention technology for the control of foodborne pathogens, especially on the surfaces of beef and pork carcasses . The results of this study also suggest that an extension of the shelf life of meat can result from the decreased growth of psychrotrophic spoilage organisms. Biochim Biophys Acta, 2003 Mar 17, 1631(2), 188 - 96 The polar lipid composition of Mesorhizobium ciceri; Choma A et al.; The extractable lipid composition of Mesorhizobium ciceri strain HAMBI 1750 grown in a phosphate sufficient medium (79CA) is reported . Cardiolipin (CL-27% of total lipids), phosphatidylglycerol (PG-18%), phosphatidylethanolamine (PE-1%), phosphatidylcholine (PC-30%) and two methylated derivatives of PE, i.e . phosphatidyl-N, N-dimethylethanolamine (DMPE-1%) and phosphatidyl-N-monomethylethanolamine (MMPE-1%), were found to make up the phospholipids of the analysed bacteria . Nonphosphorus, ornithine-containing lipid (OL-10%) was also detected . Polar groups of phospholipids were predominantly acylated with cis-11,12-methyleneoctadecanoyl (lactobacillic) residues, whereas the ornithine lipid contained mainly 3-hexadecanoyloxy-11,12-methyleneoctadecanoic acid bound to the alpha-amino group. Stomatologiia (Mosk), 2002, 81(6), 53 - 6 {Preoperative bifidum-lactobacterin therapy in children with cleft lip and palate}; Murtazaev SM et al.; Microbiological and immunologic investigations of peripheric blood in 125 children were conducted . They determined a positive role of early correction with eubiotics (bifidum-lactobacterium) in children with congenital cleft lip and congenital cleft palate during 8-9 months before chiloplasty and uraniscoplasty . Using eubiotics results in normalization and restoration of the intestinal microflora, which leads to restoration of cellular and humoral immunity and to decreased rates of accompanying and postoperative complications. Eur J Biochem, 2003 Mar, 270(6), 1102 - 16 Outer sphere mutagenesis of Lactobacillus plantarum manganese catalase disrupts the cluster core . Mechanistic implications; Whittaker MM et al.; X-ray crystallography of the nonheme manganese catalase from Lactobacillus plantarum (LPC) {Barynin, V.V., Whittaker, M.M., Antonyuk, S.V., Lamzin, V.S., Harrison, P.M., Artymiuk, P.J . & Whittaker, J.W . (2001) Structure9, 725-738} has revealed the structure of the dimanganese redox cluster together with its protein environment . The oxidized {Mn(III)Mn(III)} cluster is bridged by two solvent molecules (oxo and hydroxo, respectively) together with a micro 1,3 bridging glutamate carboxylate and is embedded in a web of hydrogen bonds involving an outer sphere tyrosine residue (Tyr42) . A novel homologous expression system has been developed for production of active recombinant LPC and Tyr42 has been replaced by phenylalanine using site-directed mutagenesis . Spectroscopic and structural studies indicate that disruption of the hydrogen-bonded web significantly perturbs the active site in Y42F LPC, breaking one of the solvent bridges and generating an 'open' form of the dimanganese cluster . Two of the metal ligands adopt alternate conformations in the crystal structure, both conformers having a broken solvent bridge in the dimanganese core . The oxidized Y42F LPC exhibits strong optical absorption characteristic of high spin Mn(III) in low symmetry and lower coordination number . MCD and EPR measurements provide complementary information defining a ferromagnetically coupled electronic ground state for a cluster containing a single solvent bridge, in contrast to the diamagnetic ground state found for the native cluster containing a pair of solvent bridges . Y42F LPC has less than 5% of the catalase activity and much higher Km for H2O2 ( approximately 1.4 m) at neutral pH than WT LPC, although the activity is slightly restored at high pH where the cluster is converted to a diamagnetic form . These studies provide new insight into the contribution of the outer sphere tyrosine to the stability of the dimanganese cluster and the role of the solvent bridges in catalysis by dimanganese catalases. J Appl Mi