J Appl Microbiol, 2000 Jan, 88(1), 176 - 82 Effect of pH control on lactic acid fermentation of starch by Lactobacillus manihotivorans LMG 18010T; Guyot JP et al.; Lactic acid fermentation of starch by Lactobacillus manihotivorans LMG 18010T, a new amylolytic L(+) lactic acid producer, was investigated and compared with starch fermentation by Lact . plantarum A6 . At non-controlled pH, growth and lactic acid production from starch by Lact . manihotivorans LMG 18010T lasted 25 h . Specific growth and lactic acid production rates continuously decreased from the onset of the fermentation, unlike Lact . plantarum A6 which was able to grow and convert starch product hydrolysis into lactic acid more rapidly and efficiently at a constant rate up to pH 4.5 . In spite of complete and rapid starch hydrolysis by Lact . manihotivorans LMG 18010T during the first 6 h, only 45% of starch hydrolysis products were converted to lactic acid . When pH was maintained at 6.0, lactic acid, amylase and final biomass production by Lact . manihotivorans LMG 18010T increased markedly and the fermentation time was reduced by half . Under the same conditions, an increase only in amylase production was observed with Lact . plantarum A6 . When grown on glucose or starch at pH 6.0, Lact . manihotivorans LMG 18010T had an identical maximum specific growth rate (0.35 h(-1)), whereas the maximum rate of specific lactic acid production was three times higher with glucose as substrate . Lactobacillus manihotivorans LMG 18010T did not produce amylase when grown on glucose . Based on the differences in the physiology between the two species and other amylolytic lactic acid bacteria, different applications may be expected. J Appl Microbiol, 2000 Jan, 88(1), 142 - 53 Effect of some nutritional and environmental parameters on the production of diacetyl and on starch consumption by Pediococcus pentosaceus and Lactobacillus acidophilus in submerged cultures; Escamilla ML et al.; Three series of 5-day submerged cultures with Pediococcus pentosaceus MITJ-10 and Lactobacillus acidophilus Hansen 1748 were carried out in starch-based media, and the effect of cultural factors on the changes of starch, diacetyl and amylase activity determined . In axenic cultures, Ped . pentosaceus MITJ-10 produced more diacetyl (63.27 mg l(-1)) by adding glucose, yeast extract and CaCO3 (P < 0.01), at 28 degrees C (P < 0.05); but more starch was consumed (18.4 g l(-1)) in the absence of glucose (P < 0.01) . Lact . acidophilus Hansen 1748 consumed more starch (26.56 g l(-1)) at 28 degrees C, with CaCO3, glucose (P < 0.01) and yeast extract (P < 0.05); however, the amylolytic activity (10077U l(-1)) was favoured at 35 degrees C (P < 0.01) . Little starch was consumed in mixed cultures due to the low pH; nevertheless, diacetyl content rose to 135.76 mg l(-1) at 32 degrees C (P < 0.01) . Therefore, both studied strains might be useful to produce aromatic extensors from starchy substrates . These natural aromatic extensors are of interest to the food industry. J Appl Microbiol, 2000 Jan, 88(1), 81 - 9 Purification and some characteristics of enterocin ON-157, a bacteriocin produced by Enterococcus faecium NIAI 157; Ohmomo S et al.; Bacteriocin-like activity (BLA) was screened in 690 strains of lactic acid bacteria isolated from plant materials such as silages and fermented vegetables . Among them, a strain identified as Enterococcus faecium NIAI 157 showed a clear BLA against the indicator strain, Ent . faecium IFO 13712 . The proteinaceous nature and antimicrobial activity against closely related species strongly indicated that this BLA was a bacteriocin and was designated enterocin ON-157 . The bacteriocin activity of this strain was extracellularly produced in the logarithmic growth phase in MRS broth and purified by ultrafiltration, ammonium sulphate precipitation and cation-exchange chromatography . Purified enterocin ON-157 had a molecular weight of approximately 2500 Da in SDS-PAGE analysis and was easily inhibited by treatment with alpha-amylase and proteolytic enzymes . Enterocin ON-157 had a bactericidal mode of action and inhibited the growth of the enterococci, Lactobacillus sake and Listeria monocytogenes . Enterococcus faecium NIAI 157 harboured two plasmids, 49.0 kb and 43.7 kb, and a variant missing a larger plasmid by curing with novobiocin lost the bactriocin activity. J Appl Microbiol, 2000 Jan, 88(1), 58 - 68 The microbial association of Greek taverna sausage stored at 4 and 10 degrees C in air, vacuum or 100% carbon dioxide, and its spoilage potential; Samelis J et al.; Strains of the Lactobacillus sakei/curvatus group, mainly non-slime-producing Lact . sakei, dominated the microbial flora of industrially manufactured taverna sausage, a traditional Greek cooked meat, stored at 4 degrees C and 10 degrees C in air, vacuum and 100% CO2 . Atypical, arginine-positive and melibiose-negative strains of this group were isolated . The isolation frequency of Lact . sakei/curvatus from sausages stored anaerobically was as high as 92-96%, while other meat spoilage organisms were practically absent . Conversely, in air-stored sausages, leuconostocs, mainly Leuconostoc mesenteroides ssp . mesenteroides, had a considerable presence (14-21%), whereas Brochothrix thermosphacta, pseudomonads and Micrococcaceae grew, but failed to increase above 10(5) cfu g(-1) in all samples during storage . Only yeasts were able to compete against LAB and reached almost 10(7) cfu g(-1) after 30 d of aerobic storage at 10 degrees C . The great dominance (> 10(8) cfu g(-1)) of LAB caused a progressive decrease of pH and an increase of the concentration of L-lactate, D-lactate and acetate in all sausage packs . The growth of LAB and its associated chemical changes were more pronounced at 10 degrees C than 4 degrees C . At both storage temperatures, L-lactate and acetate increased more rapidly and to a higher concentration aerobically, unlike D-lactate, which formed in higher amounts anaerobically . Storage in air was the worst packaging method, resulting in greening and unpleasant off-odours associated with the high acetate content of the sausages . Carbon dioxide had no significant effect on extending shelf-life . The factors affecting the natural selection of Lact . sakei/curvatus in taverna sausage are discussed . Moreover, it was attempted to correlate the metabolic activity of this group with the physicochemical changes and the spoilage phenomena occurring in taverna sausage under the different storage conditions. J Appl Microbiol, 2000 Jan, 88(1), 52 - 7 The growth of Listeria monocytogenes in cheese packed under a modified atmosphere; Whitley E et al.; The effect of modified atmosphere Packaging (MAP) on the growth of Listeria monocytogenes in mould ripened cheeses was studied at refrigeration temperatures (2-8.3 degrees C) over a storage period of 6 weeks . Control experiments in cling film with no atmospheric modification produced a lag time before growth of up to 1 week and rapid subsequent growth . MAP with a CO2 concentration of less than 20% allowed growth to occur but when O2 was incorporated; the lag time was reduced from 3 to 2 weeks and subsequent growth was also faster, producing an increase in cell numbers of 1.4 log cycles over the incubation period . N2-MAP in the absence of O2 increased the lag time to 3 weeks and slowed growth, while the inclusion of CO2 extended the lag to 3 weeks and slowed subsequent growth even more . In MAP with 80:10:10 (v/v/v) N2:CO2:O2, there was a lag period of 2-3 weeks before growth of L . monocytogenes occurred, while the total viable aerobic count (TVAC) decreased by 2-3 log cycles and the total Lactobacillus count showed little change . It was concluded that MAP was not suitable for preventing the growth of L . monocytogenes in such cheeses. J Appl Microbiol, 2000 Jan, 88(1), 44 - 51 Bacteriocin production by lactic acid bacteria isolated from Rioja red wines; Navarro L et al.; Forty-two lactic acid bacteria (LAB) of the genera Lactobacillus (32), Leuconostoc (6), Pediococcus (3) and Lactococcus (1), isolated from Rioja red wines, were tested for antimicrobial activity . All these strains, as well as 18 Leuconostoc oenos and 19 yeast strains were used as indicators . Only nine strains showed antimicrobial activity, and all were of the species Lactobacillus plantarum, which constitutes the predominant microflora in Rioja red wines after alcoholic fermentation . Lact . plantarum strain J-51 showed the widest range of action, inhibiting the growth of 31 strains of the four studied LAB genera . Lact . plantarum J-51 antimicrobial activity was lost after treatment with proteases, suggesting a proteinaceous nature for this activity . It was found to be stable between pH 3 and 9 and under strong heating conditions (100 degrees C for 60 min) . Polymerase chain reaction (PCR) analysis of Lact . plantarum J-51 genome revealed the presence of the plnA gene that encodes the plantaricin precursor PlnA . A 366-bp fragment was sequenced and showed 95% identity with pln locus of Lact . plantarum C-11 . The deduced precursor peptide sequence showed one mutation (Gly7 to Ser7) at the double glycine leader peptide, and the three putative 26-, 23- and 22-residue active peptides remain identical to those of Lact . plantarum C-11 . Therefore, antimicrobial peptides constitute a potent adaptation advantage for those strains that dominate in a medium such as wine, and can play an important role in the ecology of wine microflora. Acta Microbiol Immunol Hung, 2000, 47(1), 53 - 61 Microbial study of farmhouse ewe cheese during storage in olive oil; Gomez R et al.; The effect of storing farmhouse ewe cheese in oil and in vacuo over long periods of time on physicochemical properties (water activity and pH) and the microbiota of the cheese was investigated . The storage conditions were found to scarcely influence the sample pH . Also, the initial water activity (a(w) = 0.961) and its value after 9 months of storage (0.927) were both very similar to those for naturally ripened cheese . The incidence of pathogenic microbial groups was found to decrease with storage time (counts fell below 1 log CFU/g) . The flora that effects proteolytic changes in cheese consisted of lactic microorganisms (viz . lactococci and lactobacilli), in addition, after 6 months of storage, of enterococci . The last are responsible for the formation of large amounts of soluble nitrogen (SN), non-protein nitrogen (NPN) and aminoacid nitrogen (NH2-N), which provide this type of cheese with very special sensory features while preventing dehydration and thus lengthening its shelf life. Bioseparation, 1999, 8(1-5), 229 - 35 On-line monitoring of glucose and/or lactate in a fermentation process using an expanded micro-bed flow injection analyser; Nandakumar MP et al.; A novel flow injection biosensor system for monitoring fermentation processes has been developed using an expanded micro bed as the enzyme reactor . An expanded bed reactor is capable of handling a mobile phase containing suspended matter like cells and cell debris . Thus, while the analyte is free to interact with the adsorbent, the suspended particulate matter passes through unhindered . With the use of a scaled down expanded bed in the flow injection analysis (FIA) system, it was possible to analyse samples directly from a fermentor without the pretreatment otherwise required to extract the analyte or remove the suspended cells . This technique, therefore, provides a means to determine the true concentrations of the metabolites in a fermentor, with more ease than possible with other techniques . Glucose oxidase immobilised on STREAMLINE was used to measure glucose concentration in a suspension of dead yeast cells . There was no interference from the cell particles even at high cell densities such as 15 gm dry weight per litre . The assay time was about 6 min . Accuracy and reproducibility of the system was found to be good . In another scheme, lactate oxidase was covalently coupled to STREAMLINE for expanded bed operation . With the on-line expanded micro bed FIA it was possible to follow the fermentation with Lactobacillus casei. Bioseparation, 1999, 8(1-5), 159 - 68 Expanded bed adsorption as a unique unit operation for the isolation of bacteriocins from fermentation media; Callewaert R et al.; Expanded bed adsorption using a strong cation exchanger allowed the direct isolation of amylovorin L471, a bacteriocin from Lactobacillus amylovorus DCE 471, from the fermentation medium . The pH of the loading and elution buffer were optimised in a packed bed with cell-free culture supernatant . Bound bacteriocin was eluted with 1.0 M NaCl . The highest recovery (30%) was obtained at the lowest pH (3.6) . At higher pH values the recovery was lower, namely 12%, 15% and 7% at pH 4.5, 6.5 and 8.0, respectively . In expanded bed mode, direct isolation of the bacteriocin from the fermentation medium at pH 3.6 (loading and elution) initially resulted in a recovery of 12% . After optimisation of the pH (loading and elution at pH 3.6 and 6.5, respectively), the recovery for amylovorin L471 increased up to 30% and higher . Recovery of enterocin A from Enterococcus faecium CTC 492 fermentation medium averaged 15% (loading and elution at pH 3.6 and 6.0, respectively) . With pediocin, produced by Pediococcus acidilactici ATCC 8042, 26% recovery was obtained at a pH of 6.5 during loading and elution . Low recoveries can be ascribed to non-optimal operation conditions (pH of loading and elution buffer), inactivation of the bacteriocin on a cationic resin, and the formation of more insoluble and less active, strongly hydrophobic bacteriocin aggregates upon further purification. Int J Food Microbiol, 1999 Nov 15, 52(3), 155 - 61 Arginine, citrulline and ornithine metabolism by lactic acid bacteria from wine; Arena ME et al.; The catabolism of arginine, an amino acid found in grape juice and wine, citrulline and ornithine was investigated in four lactic acid bacteria . Only Lactobacillus hilgardii X1B catabolized arginine and excreted citrulline into the medium . The recovery of arginine as ornithine was lower than the expected theoretical value . The arginase-urease pathway was not detected indicating that the amino acid degradation was carried out only by the arginine dihydrolase pathway . Oenococcus oeni m, a strain not able to utilize arginine, degraded citrulline that was completely recovered as ornithine, ammonia and CO2 . Lactobacillus hilgardii X1B catabolized citrulline but it was only 44% recovered as ornithine . The citrulline utilization by Oenococcus oeni m may be important for two reasons: it can gain extra energy for growth from citrulline metabolism, and the amino-acid diminution could avoid the possibility of ethyl carbamate formation from the citrulline naturally present in wine. FEMS Microbiol Lett, 2000 Apr 1, 185(1), 1 - 7 Enterococcal-type glycopeptide resistance genes in non-enterococcal organisms; Patel R; Although the emergence of vancomycin-resistant enterococci can be attributed, in part, to the increasing use of vancomycin in clinical practice, and glycopeptide use in animal husbandry, the origins of the enterococcal vancomycin resistance genes are not clear . The vancomycin resistance-associated genes in Enterococcus gallinarum, Enterococcus casseliflavus/flavescens, Lactobacillus spp., Leuconostoc spp., Pediococcus spp., and Erysipelothrix rhusiopathiae, are not the source of the high-level vancomycin resistance-associated genes in enterococci . There are, however, environmental organisms which have been found to have gene clusters homologous to the enterococcal vanA, vanB and vanC gene clusters; these include the biopesticide Paenibacillus popilliae, and, to a lesser extent, the glycopeptide-producing organisms Amycolatopsis orientalis and Streptomyces toyocaensis . Still, the exact sources of the enterococcal vancomycin resistance genes remain a mystery. Akush Ginekol (Sofiia), 1999, 38(2), 21 - 3 {Streptococcus group B isolated in 3 microscopic displays from the vaginal secretions of pregnant women}; Shopova E et al.; Over a period of 20 months we investigated 1366 vaginal specimens from pregnant women for GBS (group B streptococci) carrier in three microscopic patterns, evaluated by Nugent score system . More frequently we isolated GBS in group I intermedia (score 4-6)--20.8%, (when Lactobacillus spp . is missing--67.4%, without or associated with other nonanaerobic microorganisms (60.5%) . All isolated GBS strains showed sensitivity to ampicillin and carbenicillin . Good sensitivity was found to cefazolin (92.4%) ant to cefuroxim (94.9%) . The strains showed 13.7% resistance to erythromycin and 4.1% to clindamycin. Arch Gynecol Obstet, 1999 Nov, 263(1-2), 34 - 6 Accuracy of rapid antigen detection test for group B streptococci in the indigenous vaginal bacterial flora; Donders GG et al.; OBJECTIVE: To test the sensitivity of the rapid group B streptococci (GBS) antigen test ICONR and compare its accuracy in women with vaginal enterococci or with non-specific disturbance of the lactobacillary flora . STUDY DESIGN: The ICONR, aerobic culture and a microscopic wet mount evaluation were done on a vaginal sample in 254 unselected women presenting for routine gynecologic care in an academic hospital in Flanders, Belgium and tested by Chi2 {diagnostic odds ratio (DOR) and its 95 percent confidence limits} . RESULTS: Sensitivity of the test was 70%, specificity 99.5% . Prevalence of GBS was 10.6% overall, 23% in the group with abnormal vaginal flora and 7% in the normal group (p=0.002) . Accuracy of the ICONR was not affected by abnormal vaginal flora, but was significantly lower in the presence of enterococci: the DOR decreased from 490 to 58, and the positive predictive value from 94 to 80% . CONCLUSION: With a sensitivity of 70% the enzyme immunoassay ICONR does not appear to be suitable as a practical screening tool for detecting GBS carriage in normal or preterm laboring women . In the presence of enterococci the test performed less well, with a DOR falling by 8 to 9 fold . We presume this is due to lower specificity in vivo in the presence of enterococci, as non-specific disturbance of the lactobacillary flora did not interfere with test results. Int J STD AIDS, 2000 Mar, 11(3), 150 - 5 Diagnosis of bacterial vaginosis by wet mount identification of bacterial morphotypes in vaginal fluid; Schmidt H et al.; In order to develop a more practical way of diagnosing bacterial vaginosis (BV), we evaluated a scoring system, weighting small bacterial morphotypes versus lactobacillary morphotypes in wet mounts, assessed criteria for BV and normalcy from this scoring, and then evaluated their reproducibility and accuracy . We examined 754 women for pH, homogeneous vaginal discharge, amine odour, clue cells and the composite clinical diagnosis . We also examined wet mounts for small bacterial morphotypes and lactobacillary morphotypes, and weighted their quantitative presence as a bacterial morphotype score . The term 'small bacterial morphotypes' denotes a group of small bacillary forms comprising coccobacilli, tiny rods, and mobile curved rods . The different characteristics of BV were all gradually associated with increased bacterial morphotype scoring . We deemed a score of 0-1 as normal, 2-4 as intermediate phase, grade I, 5-6 as intermediate phase, grade II, and 7-8 indicative of BV . Reproducibility of the interpretation was high, both for the new grading system (weighted Kappa 0.90 in women perceiving and 0.81 in women not perceiving abnormal vaginal discharge) and for the new criterion for BV (non-weighted Kappa 0.91 and 0.84 in the 2 groups of women) . The new criterion also proved highly concurrent with the composite clinical diagnosis (Kappa 0.91 and 0.81 in the 2 groups) . In conclusion, the wet mount bacterial morphotype scoring is valid for grading of the disorder of the vaginal microbial ecosystem, and the new criterion for BV a more practical option than existing diagnostic methods. J Immunol, 2000 Apr 1, 164(7), 3733 - 40 Lactobacilli and Streptococci activate NF-kappa B and STAT signaling pathways in human macrophages; Miettinen M et al.; Gram-positive bacteria induce the production of several cytokines in human leukocytes . The molecular mechanisms involved in Gram-positive bacteria-induced cytokine production have been poorly characterized . In this work we demonstrate that both nonpathogenic Lactobacillus rhamnosus GG and pathogenic Streptococcus pyogenes (group A streptococci) induce NF-kappa B and STAT DNA-binding activity in human primary macrophages as analyzed by EMSA . NF-kappa B activation was rapid and was not inhibited by a protein synthesis inhibitor cycloheximide, suggesting that these bacteria could directly activate NF-kappa B . STAT1, STAT3, and IFN regulatory factor-1 DNA binding was induced by both bacteria with delayed kinetics compared with NF-kappa B . In addition, streptococci induced the formation of IFN-alpha-specific transcription factor complex and IFN-stimulated gene factor-3 (ISGF3) . STAT1 and STAT3 activation and ISGF3 complex formation were inhibited by cycloheximide or by neutralization with IFN-alpha/beta-specific Abs . Streptococci were more potent than lactobacilli in inducing STAT1, ISGF3, and IFN regulatory factor-1 DNA binding . Accordingly, only streptococci induced IFN-alpha production . The activation of the IFN-alpha signaling pathway by streptococci could play a role in the pathogenesis of these bacteria . These results indicate that extracellular Gram-positive bacteria activate transcription factors involved in cytokine signaling by two mechanisms: directly, leading to NF-kappa B activation, and indirectly via cytokines, leading to STAT activation. Gene, 2000 Jan 25, 242(1-2), 155 - 66 The genetic switch for the regulatory pathway of Lactobacillus plantarum phage (phi)g1e: characterization of the promoter P(L), the repressor gene cpg, and the cpg-encoded protein Cpg in Escherichia coli; Kakikawa M et al.; The structural and functional features of the approximately 530 bp P(L)/Gb5-Gb6-cpg-Gb7 region (P(L) overlaps Gb5) for the lysogenic pathway of L . plantarum phage (phi)gle were investigated using the cat gene of E . coli plasmid pKK232-8 as a reporter . In E . coli XL1-Blue, a recombinant plasmid pKPL2 (cat under P(L)/Gb5-Gb6) exhibited distinct CAT activity, whereas the activity of pKPLCP1 (cat under P(L)/Gb5-Gb6-cpg) was only marginal . When pKPL2 was coexistent with a compatible derivative of plasmid pACYC177 carrying P(L)/Gb5-Gb6-cpg, the CAT activity was declined to the level of pKPLCP1 . On the other hand, the cpg-encoded protein Cpg was overproduced in E . coli under P(T7) . The molecular mass of the purified Cpg (14.5 kDa on a SDS gel) corresponded well with that (15.1 kDa) predicted from the DNA sequence . Gel-shift and footprinting assays demonstrated that Cpg selectively binds to about 25 bp bases centered around the GATAC-box (from 1 to 7) . Moreover, protein crosslinking experiments using glutaraldehyde showed that Cpg most likely functions as a dimeric form . Thus, the present results indicate that Cpg probably represses P(L) through binding to the operator GATAC-box(es), and the P(L)/cpg region might participate in the lysogenic pathway. J Food Prot, 2000 Mar, 63(3), 412 - 4 Binding of aflatoxin B1 alters the adhesion properties of Lactobacillus rhamnosus strain GG in a Caco-2 model; Kankaanpaa P et al.; Lactic acid bacteria have been previously reported to possess antimycotoxigenic activities both in vitro and in vivo . The objective of this study was to investigate the effect of aflatoxin B1 on adhesion capability of Lactobacillus rhamnosus strain GG using a Caco-2 adhesion model . Removal of aflatoxin B1 by L . rhamnosus strain GG reduced the adhesion capability of this strain from 30% to 5% . It is therefore concluded that aflatoxins may influence the adhesion properties of probiotics able to sequester them, and subsequently these bacteria may reduce the accumulation of aflatoxins in the intestine via increased excretion of an aflatoxin-bacteria complex. Plant Foods Hum Nutr, 1999, 54(3), 239 - 50 Effect of process improvement on the physico-chemical properties of infant weaning food from fermented composite blends of cereal and soybeans; Onilude AA et al.; Saccharomyces cerevisiae and Lactobacillus plantarum ATCC 10776 were used as starters to ferment various composite blends of cereals and legumes produced through malting and toasting of two varieties of maize--Zea mays (DMR-LSR white & DMR-ESR yellow), sorghum--Sorghum bicolor (Dawa white & Dawa red) and one variety of soybeans (Glycine ax) . Compared to the untoasted and unmalted fermented blends, a relatively lower pH (3.6), highly sour product was obtained with 12 h of fermentation . Results also showed that cereal and soybean toasting brought about a better reconstitution indices (B25, 84 ml; B45, 87 ml), water holding capacities (B25, 0.68 ml/g; B45, 0.62 ml/g), bulk densities (C15, 11.6; C35, 10.8) and gross energy (B15, 501.5 k cal/100g; B45, 508.5 kcal/100g) at the end of fermentation . Furthermore, reductions in total polyphenol and tannin contents were observed with fermentation of toasted and malted cereal blends supplemented with toasted and malted soybeans while porridges from the same blends displayed desirable starch stability and consistent gelling tendency, although B15 (a ferment of malted, toasted white maize supplemented with toasted and malted soybean) fell within acceptable limits . In all, the physical characteristics were affected by varieties of cereal and soybeans. Enzyme Microb Technol, 2000 Mar 1, 26(5-6), 431 - 445 Comparison of simple neural networks and nonlinear regression models for descriptive modeling of Lactobacillus helveticus growth in pH-controlled batch cultures; Schepers AW et al.; A set of 20 Lactobacillus helveticus growth curves was obtained from pH-controlled batch cultures with different pH setpoints, whey permeate and yeast extract concentrations . To find the best descriptive model of the biomass concentration versus time (y = X(t)) growth curve, fitting results of a large number of models were compared with statistical and approximate methods . Models studied included simple neural networks, reparameterized Logistic, Gompertz, Richards, Schnute, Weibull, and Morgan-Mercier-Flodin models, Amrane-Prigent model, and four new models based on autonomous growth functions . Simple neural networks with only four weights were good descriptive models of the growth curves and fitting qualities were similar to those of the best existing four-parameter models, such as the Logistic model . However, meaningful parameters had to be calculated numerically and use of simple neural networks yielded no distinctive advantages over other models . A new five-parameter model, based on an autonomous growth function, yielded the best fitting results, even when the number of model parameters was accounted for in the comparisons . However, the maximum specific growth rate was not always well estimated . Therefore the five-parameter Richards model was chosen as the best descriptive model of the growth curve. J Reprod Med, 2000 Feb, 45(2), 131 - 4 Trichomonas vaginalis and bacterial vaginosis . Coexistence in vaginal wet mount preparations from pregnant women; Franklin TL et al.; OBJECTIVE: To identify how frequently trichomoniasis and characteristics of bacterial vaginosis (BV) occur concomitantly in wet mount preparations from pregnant women . STUDY DESIGN: Diagnosis of trichomoniasis was predicted on visualization of the organism . Diagnosis of BV required a positive volatile (whiff) test, presence of "clue cells" and one of two minor criteria: (1) absence of lactobacilli, or (2) a pH > 4.5 . Pregnant women from January 1995 to July 1997 at our clinic had wet mount/KOH preparations performed as standard prenatal care . Corresponding medical charts were analyzed for symptoms, race, BV, sexually transmitted diseases, urinary tract infections and other infections . RESULTS: Of 191 pregnant women identified, 69 had trichomoniasis . Seventy-nine percent of the 69 were African American . Fifteen percent of pregnant women (17) had concomitant trichomoniasis and BV . Irrespective of race, 35-38% of pregnant women with trichomoniasis had another sexually transmitted disease or a urinary tract infection diagnosed in that pregnancy . CONCLUSION: BV, or bacteria excess syndrome, is a frequent coinfection in pregnant women harboring Trichomonas vaginalis. Appl Microbiol Biotechnol, 2000 Feb, 53(2), 159 - 66 Influence of complex nutrients, temperature and pH on bacteriocin production by Lactobacillus sakei CCUG 42687; Aasen IM et al.; The effects of process conditions and growth kinetics on the production of the bacteriocin sakacin P by Lactobacillus sakei CCUG 42687 have been studied in pH-controlled fermentations . The fermentations could be divided into phases based on the growth kinetics, phase one being a short period of exponential growth, and three subsequent ones being phases of with decreasing specific growth rate . Sakacin P production was maximal at 20 degrees C . At higher temperatures (25-30 degrees C) the production ceased at lower cell masses, when less glucose was consumed, resulting in much lower sakacin P concentrations . With similar media and pH, the maximum sakacin P concentration at 20 degrees C was seven times higher than that at 30 degrees C . The growth rate increased with increasing concentrations of yeast extract, and the maximum concentration and specific production rate of sakacin P increased concomitantly . Increasing tryptone concentrations also had a positive influence upon sakacin P production, though the effect was significantly lower than that of yeast extract . The maximum sakacin P concentration obtained in this study was 20.5 mg l(-1) . On the basis of the growth and production kinetics, possible metabolic regulation of bacteriocin synthesis is discussed, e.g . the effects of availability of essential amino acids, other nutrients, and energy. J Otolaryngol, 2000 Feb, 29(1), 28 - 34 Clinical evaluation of a commercially available oral moisturizer in relieving signs and symptoms of xerostomia in postirradiation head and neck cancer patients and patients with Sjögren's syndrome; Rhodus NL et al.; A major complication of irradiation therapy for head and neck cancer is salivary gland dysfunction and xerostomia . The purpose of this clinical investigation was to evaluate the effects of a commercially available oral moisturizer (Optimoist) on salivary flow rate, symptoms of xerostomia, oral pH, oral microflora, and swallowing in postirradiation head and neck cancer patients (XRT) and patients with Sjogren's syndrome (SS) . Subjects who were post-XRT and subjects with SS (n = 24; mean age = 54.1) discontinued their use of any salivary substitute or moisturizer for 2 weeks prior to entering the study . Baseline whole unstimulated saliva was collected for 5 minutes using a standard sialometric technique . Candida albicans and Lactobacillus cultures were performed using kits from Orion Diagnostica, Inc., and a pH analysis was performed on the salivary sample using a Markson (model 00663) pH meter . Swallowing was assessed by clinical measures by videofluoroscopic techniques . Several subjective assessments were performed to evaluate symptoms of xerostomia . Subjects were instructed in the use of a daily diary and to use only the provided article ad libitum for a period of 2 weeks . After the 2-week period, the results indicated significant subjective and objective improvements in signs and symptoms of xerostomia . Whole unstimulated salivary flow rate improved from (mean +/- SEM) 0.1150 +/- 0.02 to 0.2373 +/- 0.09 mL/min . Salivary pH did not change . Global subjective improvement in xerostomia improved in 58% of the subjects . Candida colonization decreased in 43% of the subjects . There was no change in Lactobacilli colonization . Swallowing objectively improved in 75% of subjects . These results indicate significant improvement in both signs of hyposalivation and symptoms of xerostomia with the use of Optimoist in postirradiation head and neck cancer patients and patients with SS. Mikrobiol Z, 1999 Nov-Dec, 61(6), 42 - 50 {The bacteriocinogenic and lysozyme-synthesizing activity of lactobacilli}; Kovalenko NK et al.; It was shown that lactic acid bacteria isolated from different sources produced along with lactic acid the bacteriocine substances . Strains producing bacteriocines were mainly isolated from digestive tract of people and calves . Bacteriogenic properties have been shown more frequently in the case of lactobacilli (7.8%) more rarely--in coccoid forms of lactic acid bacteria (4.5%) . At the same time bacteriocine activity was higher in the case of Enterococcus and depended both on the strain type and cultivation conditions . Synthesis of bacteriocines started from the hours of phase of the culture growth and reached maximum at the beginning of stationary phase . The bacteriocine accumulation rate depended on composition of the growth medium. Eur J Clin Microbiol Infect Dis, 2000 Jan, 19(1), 33 - 8 Comparison of a commercial disk test with vancomycin and colimycin susceptibility testing for identification of bacteria with abnormal gram staining reactions; Fenollar F et al.; In an effort to identify bacteria that fail to give the expected Gram reaction, thus leading to misidentification, two nonstaining tests for Gram reaction, vancomycin and colimycin susceptibility testing and the Gram-Sure test (Remel, USA), were employed on 145 strains from 42 gram-negative and gram-positive genera with contradictory Gram stain results . The Gram-Sure test is a commercially available disk that detects the presence of L-alanine-aminopeptidase, an enzyme usually found only in the cell wall of gram-negative bacteria . In this test, aminopeptidase activity is detected using a substrate that can be hydrolyzed to produce a fluorescent compound under long-wave UV light . The commercial disk test and vancomycin plus colimycin susceptibility testing appeared to perform equally well except in the identification of Erysipelothrix and Lactobacillus, for which the commercial disk test was better, and Moraxella, for which vancomycin and colimycin susceptibility testing was more helpful . An advantage of the commercial disk test is that it can be performed in 10 min, whereas vancomycin and colimycin susceptibility testing requires at least 18 h . The commercial disk test is also less expensive than vancomycin and colimycin susceptibility testing . However, since the same results can be obtained with the 5 microg and 30 microg vancomycin disks, it is possible to use only one vancomycin disk, with the cost then being equivalent to that of the commercial disk test . The major inconvenience of the commercial disk test is the requirement of a UV ray . However, this test could be a useful tool for the identification of unusual organisms. Biochemistry, 2000 Mar 14, 39(10), 2429 - 35 Effects of subunit occupancy on partitioning of an intermediate in thymidylate synthase mutants; Variath P et al.; Experimental evidence for a 5-exocyclic methylene-dUMP intermediate in the thymidylate synthase reaction was recently obtained by demonstrating that tryptophan 82 mutants of the Lactobacillus casei enzyme produced 5-(2-hydroxyethyl)thiomethyl-dUMP (HETM-dUMP) (Barret, J . E., Maltby, D . A., Santi, D . V., and Schultz, P . G . (1998) J . Am . Chem . Soc . 120, 449-450) . The unusual product was proposed to emanate from trapping of the intermediate with beta-mercaptoethanol in competition with hydride transfer from H(4)folate to form dTMP . Using mutants of the C-terminal residue of thymidylate synthase, we found that the ratio of HETM-dUMP to dTMP varies as a function of CH(2)H(4)folate concentration . This observation seemed inconsistent with the conclusion that both products arose from a common intermediate in which CH(2)H(4)folate was already bound to the enzyme . The enigma was resolved by a kinetic model that allowed for differential partitioning of the intermediate formed on each of the two subunits of the homodimeric enzyme in forming the two different products . With three C-terminal mutants of L . casei TS, HETM-dUMP formation was consistent with a model in which product formation occurs upon occupancy of the first completely bound subunit, the rate of which is unaffected by occupancy of the second subunit . With one analogous E . coli TS mutant, HETM-dUMP formation occurred upon occupancy of the first subunit, but was inhibited when both subunits were occupied . With all mutants, dTMP formation occurs from occupied forms of both subunits at different rates; here, binding of cofactor to the first subunit decreased affinity for the second, but the reaction occurred faster in the enzyme form with both subunits bound to dUMP and CH(2)H(4)folate . The model resolves the apparent enigma of the cofactor-dependent product distribution and supports the conclusion that the exocyclic methylene intermediate is common to both HETM-dUMP and dTMP formation. Toxicol In Vitro, 2000 Feb, 14(1), 53 - 9 Inhibition of dihydrofolate reductase and cell growth activity by the phenanthroindolizidine alkaloids pergularinine and tylophorinidine: the in vitro cytotoxicity of these plant alkaloids and their potential as antimicrobial and anticancer agents; Rao KN et al.; The phenanthroindolizidine plant alkaloids pergularinine (PGL) and tylophorinidine (TPD) isolated from the Indian medicinal herb Pergularia pallida have been evaluated for their biological activity and assessed for the first time employing dihydrofolate reductase (DHFR) (5,6,7,8-THF: NADP(+) oxidoreductase, EC 1.5.1.3) as the probe in the present investigations . The enzyme is a key target in cancer chemotherapy and has been purified from Lactobacillus leichmannii . Cytotoxicity studies showed that both PGL and TPD are potently toxic and inhibited the growth of L . leichmannii cells (IC(50)=45 and 40 microM, respectively) . Both the alkaloids significantly inhibited DHFR activity (IC(50)=40 and 32 microM for PGL and TPD, respectively) . Alkaloid concentrations greater than 75-95 microM resulted in a complete loss of DHFR activity . Our results are suggestive of the alkaloids as potential antimicrobial and antitumour compounds . Alkaloid binding to DHFR is slow and reversible . Inhibition kinetics revealed K(i) values of 9x10(-6) M and 7x10(-6) M for PGL and TPD, respectively for the enzyme and inhibition in both the cases was a simple linear 'non-competitive' type. J Clin Microbiol, 2000 Mar, 38(3), 1302 - 4 Potential errors in recognition of Erysipelothrix rhusiopathiae; Dunbar SA et al.; Here we describe four isolations of Erysipelothrix rhusiopathiae associated with polyarthralgia and renal failure, septic arthritis, classic erysipeloid, and peritonitis . Although the biochemical identification was straightforward in each case, recognition presented a challenge to the clinical microbiologist, since in three cases E . rhusiopathiae was not initially considered due to unusual clinical presentations, in two cases the significance might not have been appreciated because growth was in broth only, and in one case the infection was thought to be polymicrobic . Because the Gram stain can be confusing, abbreviated identification schemes that do not include testing for H(2)S production could allow E . rhusiopathiae isolates to be misidentified as Lactobacillus spp . or Enterococcus spp . in atypical infections. Appl Environ Microbiol, 2000 Mar, 66(3), 1152 - 7 Antagonistic activity of Lactobacillus acidophilus LB against intracellular Salmonella enterica serovar Typhimurium infecting human enterocyte-like Caco-2/TC-7 cells; Coconnier MH et al.; To gain further insight into the mechanism by which lactobacilli develop antimicrobial activity, we have examined how Lactobacillus acidophilus LB inhibits the promoted cellular injuries and intracellular lifestyle of Salmonella enterica serovar Typhimurium SL1344 infecting the cultured, fully differentiated human intestinal cell line Caco-2/TC-7 . We showed that the spent culture supernatant of strain LB (LB-SCS) decreases the number of apical serovar Typhimurium-induced F-actin rearrangements in infected cells . LB-SCS treatment efficiently decreased transcellular passage of S . enterica serovar Typhimurium . Moreover, LB-SCS treatment inhibited intracellular growth of serovar Typhimurium, since treated intracellular bacteria displayed a small, rounded morphology resembling that of resting bacteria . We also showed that LB-SCS treatment inhibits adhesion-dependent serovar Typhimurium-induced interleukin-8 production. Appl Environ Microbiol, 2000 Mar, 66(3), 1084 - 92 Characterization and determination of origin of lactic acid bacteria from a sorghum-based fermented weaning food by analysis of soluble proteins and amplified fragment length polymorphism fingerprinting; Kunene NF et al.; The group that includes the lactic acid bacteria is one of the most diverse groups of bacteria known, and these organisms have been characterized extensively by using different techniques . In this study, 180 lactic acid bacterial strains isolated from sorghum powder (44 strains) and from corresponding fermented (93 strains) and cooked fermented (43 strains) porridge samples that were prepared in 15 households were characterized by using biochemical and physiological methods, as well as by analyzing the electrophoretic profiles of total soluble proteins . A total of 58 of the 180 strains were Lactobacillus plantarum strains, 47 were Leuconostoc mesenteroides strains, 25 were Lactobacillus sake-Lactobacillus curvatus strains, 17 were Pediococcus pentosaceus strains, 13 were Pediococcus acidilactici strains, and 7 were Lactococcus lactis strains . L . plantarum and L . mesenteroides strains were the dominant strains during the fermentation process and were recovered from 87 and 73% of the households, respectively . The potential origins of these groups of lactic acid bacteria were assessed by amplified fragment length polymorphism fingerprint analysis. Can J Microbiol, 1999 Dec, 45(12), 981 - 7 Some probiotic properties of chicken lactobacilli; Gusils C et al.; The beneficial effect of lactobacilli has been attributed to their ability to colonize human and animal gastrointestinal tracts . In this work, adhesion assays with three lactobacillus strains and intestinal fragments obtained from chickens were assessed . Lactobacillus animalis and L . fermentum were able to adhere to three kinds of epithelial cells (crop, small and large intestines) with predominance to small intestine . Among the strains considered, L . fermentum subsp . cellobiosus showed the lowest and L . animalis the highest adhesion ability . Scanning electron microphotographs showing L . animalis and L . fermentum adhering to intestinal cells were obtained . The characterization of L . animalis adhesion indicated that lectin-like structure of this strain has glucose/mannose as specific sugars of binding . However, a calcium requirement was not observed . The adhesion of L . fermentum was reduced by addition of sialic acid or mannose (P < 0.01) . These carbohydrates can be involved in the interaction between adhesin and epithelial surface . In this case, the dependence on bivalent cations was demonstrated . Lactobacillus fermentum was effective in reducing the attachment of Salmonella pullorum by 77%, while L . animalis was able to inhibit (90%, 88%, and 78%) the adhesion of S . pullorum, S . enteritidis, and S . gallinarum to host-specific epithelial fragments respectively . Our results from this in vitro model suggest that these lactobacilli are able to block the binding sites for Salmonella adhesion. J Food Prot, 2000 Feb, 63(2), 237 - 43 Reduction of biogenic amine formation using a negative amino acid-decarboxylase starter culture for fermentation of Fuet sausages; Bover-Cid S et al.; The ability of Lactobacillus sakei CTC494, a negative amino acid-decarboxylase starter culture, to reduce biogenic amine accumulation during sausage fermentation and storage at 4 and 19 degrees C was studied . The effect on the amine formation of the tyramine producer Lactobacillus curvatus CTC371, as a positive strain, was also examined in comparison to a spontaneous fermentation process without starter culture (control batch) . The polyamines spermine, spermidine, and diaminopropane were not influenced by the ripening, and their levels slightly decreased in all the batches throughout the storage . Tyramine, cadaverine, and putrescine were the main amines formed during the ripening . The addition of starter culture resulted in a decrease on the biogenic amine formation, depending on the strain inoculated . A great reduction in tyramine content was achieved when L . sakei CTC494 was inoculated, whereas L . curvatus CTC371 only attenuated tyramine accumulation compared with the control batch . Both starters were able to significantly limit the production of putrescine and cadaverine, and they inhibited tryptamine and phenylethylamine formation by the wild microbial flora . Tyramine levels of the control sausages rose during the storage at both temperatures, whereas those of cadaverine only increased at 19 degrees C . On the contrary, sausages manufactured through the starter controlled fermentation did not show changes of amine contents during the storage . The addition of a proper selected starter culture is advisable to produce safer sausages with low contents of biogenic amines. J Food Prot, 2000 Feb, 63(2), 202 - 9 The antimicrobial properties of chitosan in mayonnaise and mayonnaise-based shrimp salads; Roller S et al.; The potential for using chitosan glutamate as a natural food preservative in mayonnaise and mayonnaise-based shrimp salad was investigated . Mayonnaise containing 3 g/liter of chitosan combined with acetic acid (0.16%) or lemon juice (1.2 and 2.6%) was inoculated with log 5 to 6 CFU/g of Salmonella Enteritidis, Zygosaccharomyces bailii, or Lactobacillus fructivorans and stored at 5 and 25 degrees C for 8 days . In mayonnaise containing chitosan and 0.16% acetic acid, 5 log CFU/g of L . fructivorans were inactivated, and numbers remained below the sensitivity limit of the plate counting technique for the duration of the experiment . Z . bailii counts were also reduced by approximately 1 to 2 log CFU/g within the first day of incubation at 25 degrees C, but this was followed by growth on subsequent days, giving an overall growth delay of 2 days . No differences in counts of Z . bailii in mayonnaise stored at 5 degrees C or of Salmonella Enteritidis stored at either temperature were observed . In mayonnaise containing lemon juice at both 1.2 and 2.6%, no substantial differences were observed between the controls and the samples containing chitosan . In shrimp salads stored at 5 degrees C, the presence of a coating of chitosan (9 mg/g of shrimp) inhibited growth of the spoilage flora from approximately log 8 CFU/g in the controls to log 4 CFU/g throughout 4 weeks . However, at 25 degrees C, chitosan was ineffective as a preservative . The results demonstrated that chitosan may be useful as a preservative when combined with acetic acid and chill storage in specific food applications. J Agric Food Chem, 2000 Feb, 48(2), 239 - 44 Use of two-dimensional electrophoresis to evaluate proteolysis in salmon (Salmo salar) muscle as affected by a lactic fermentation; Morzel M et al.; Two-dimensional electrophoresis was used to study proteolysis in salmon fillets inoculated or not with the starter culture Lactobacillus sake LAD . Protein fragments appeared increasingly with time in both samples, indicating that the main quantitative changes were due to endogenous enzymes . In the most acidic zone (pI = 4-6 . 20) particularly, proteolysis was overall independent from processing . In contrast, fermentation had a significant effect in the pH range 6.20-8.35, suggesting a specificity of the bacterial proteases of L . sake toward alkaline to slightly acidic proteins . Furthermore, fragments surrounding tropomyosin (apparent pI = 4.70) appeared in fermented samples, indicating that the protein may be a suitable substrate for the metabolism of L . sake LAD. Med Microbiol Immunol (Berl), 1999 Aug, 188(1), 9 - 14 The role of tumor necrosis factor (TNF)-alpha in the antitumor effect of intrapleural injection of Lactobacillus casei strain Shirota in mice; Yasutake N et al.; The involvement of several cytokines in the antitumor effect induced by intrapleural (i.pl.) injection of heat-killed cells of Lactobacillus casei strain Shirota (LC 9018) in mice was investigated . Injection of LC 9018 i.pl . into Meth A fibrosarcoma (Meth A)-bearing mice not only significantly prolonged the survival of the mice, but also effectively inhibited the accumulation of malignant pleural fluid in the thoracic cavity . In the thoracic cavity of tumor-bearing mice treated with LC 9018, we observed large amounts of several cytokines including interleukin (IL)-1beta, interferon (IFN)-gamma, IL-12 and tumor necrosis factor (TNF)-alpha . Both anti-IFN-gamma and anti-IL-12 monoclonal antibody (mAb) treatments partially diminished the antitumor activity of LC 9018 in vivo, while the treatment of anti-IL-1beta mAb did not influence the survival of the mice . However, anti-TNF-alpha mAb treatment completely abolished the antitumor effect of LC 9018 in vivo, suggesting that in this model LC 9018 has a survival-prolonging effect involving certain cytokines . Moreover, i.pl . injection of mouse recombinant TNF-alpha into Meth A-bearing mice pretreated with anti-TNF-alpha mAb partially restored the survival-enhancing effect of LC 9018 . These results led us to conclude that TNF-alpha induced by i.pl . injection of LC 9018 plays an important role in the antitumor effect of LC 9018 in vivo. Enzyme Microb Technol, 2000 Feb 1, 26(2-4), 209 - 215 Production of lactic acid by Lactobacillus rhamnosus with vitamin-supplemented soybean hydrolysate; Kwon S et al.; Batch fermentation studies were performed to evaluate the potentials of a complex nitrogen source, soybean, as an alternative to yeast extract for the economical production of lactic acid by Lactobacillus rhamnosus . An enzyme-hydrolysate of soybean meal, Soytone, with an adequate supplementation of vitamins was found to be highly effective in supporting lactic acid production from glucose and lactose . The effects of seven selected vitamins: d-biotin, pyridoxine, p-aminobenzoic acid, nicotinic acid, thiamine, pantothenic acid, and riboflavin, on cell growth and lactic acid production were investigated to provide the basis for the optimization of vitamin supplementation to minimize the cost . Pantothenic acid was the most required compound while the other six vitamins were also essential for high lactic acid productivity . As a result of the optimization, 15 g/l yeast extract could be successfully replaced with 19.3 g/l Soytone supplemented with the vitamins, resulting in a production of 125 g/l lactic acid from 150 g/l glucose . The volumetric productivity and lactate yield were 2.27 g/l/h and 92%, respectively, which were higher than those with 15 g/l yeast extract . The raw material cost was estimated to be 21.4 cent/kg lactic acid, which was only approximately 41% of that with yeast extract. Curr Microbiol, 2000 Apr, 40(4), 245 - 9 Genetic organization and polymorphism of the guaA gene encoding the GMP synthetase in Lactobacillus rhamnosus; Grimaldi C et al.; The guaA gene encoding GMP synthetase was cloned from a potential probiotic strain of Lactobacillus rhamnosus . DNA sequence and Northern blot analysis indicated that (i) guaA did not belong to an guaAB operonic structure, conversely to enteric bacteria, (ii) L . rhamnosus guaA seemed to be highly expressed, and (iii) genetic regulation might differ from Bacillus subtilis . Moreover, differences in the genetic organization of guaA allowed discrimination of some closely related L . rhamnosus strains, with a rapid screening by PCR. Gerodontology, 1999 Jul, 16(1), 47 - 58 Caries activity and associated risk factors in elderly hospitalised population--15-months follow-up in French institutions; Guivante-Nabet C et al.; Only a few studies have been published concerning hospitalised elderly disabled people . OBJECTIVES: 1) to investigate the oral health status of elderly French patients hospitalised in the two main geriatric hospitals of Paris . 2) to describe the respective influences of general parameters (type of hospitalisation, pathologies and medication) on oral environment parameters . 3) to analyse the influences of these oral parameters on caries activity in Long-Term Care (LTCF) and in rehabilitation facilities (RF) patients and to study the incidence and the time-course of caries in these specific population . SUBJECTS: 117 subjects (mean age = 83.0 years, SD = 7.8, range = 64 to 102 years) were examined at baseline and 32 of the 50 LTCF subjects were reexamined 15-months later . METHODS: The general parameters recorded were age, gender, type of hospitalisation, period of stay, removable prosthesis, general diseases, number of diagnoses, medications with hyposalivary side-effects . The oral environment parameters recorded were flow rate, buffer capacity, mutans streptococci and lactobacilli counts, measured at baseline by tests on stimulated saliva, and plaque index . Crown and root surfaces were recorded according to a modified caries activity index . RESULTS: Among the polypathological subjects (85.5% of the population), the number of diseases ranged from 2 to 8 . The LTCF patients had a significantly higher mean number of diagnoses (3.5; SD = 1.5) than the RF patients (2.8; SD = 1.4) . 76.9% of patients were taking medications with hyposalivary side-effects . The stimulated flow rate ranged from 0.02 ml/min to 5 ml/min . Its mean was significantly lower for LTCF patients (0.67 ml/min; SD = 0.51) than for RF patients (1.12 ml/min; SD = 0.89) . The plaque index was significantly higher in LTCF subjects and in patients with mental diseases . At baseline, 17,442 crown and root surfaces were examined . Flow rate was related to crown caries and buffer capacity to root caries . During the 15-months follow-up, the mean number of active root surfaces was significantly increased: from 0.148 (SD = 0.116) at baseline vs . 0.250 (SD = 0.174) at the second examination . CONCLUSIONS: The strongest relationship in the present study between oral parameters and caries activity was the negative relationship between buffer capacity and active root caries . This study confirms an association between the type of hospitalisation and both salivary parameters flow rate and plaque index . This investigation illustrates the critical need for hygiene and oral care, in this elderly disabled population. Gerodontology, 1999 Jul, 16(1), 37 - 46 Factors which are associated with dental decay in the older individual; Loesche WJ et al.; OBJECTIVES: To improve reliability of salivary bacterial cultures as a surrogate for plaque levels of cariogenic bacterial species by reporting the salivary CFUs of these organisms as a function of the number of teeth . DESIGN: Cross-sectional collection of data in a convenience sample of adults over 60 years of age . SETTING: Hospital Dental clinic, University bacteriology laboratory . SUBJECTS: 523 older dentate subjects, average age 70, including 412 subjects who were in an independent living status and 111 in a dependent-living situation . MAIN OUTCOME MEASURES: Subjects were examined for decay and the presence of salivary factors including the levels of S . mutans, lactobacilli, yeast and other bacteria . The salivary levels of the bacteria were adjusted for the number of teeth in the mouth, and the resultant values were entered into multivariable logistic regression models along with clinical and other salivary parameters . RESULTS: Mutans streptococci levels reported as CFUs/ml saliva per tooth were significantly associated with coronal surface decay, and lactobacilli, reported in a similar way, were significantly associated with root surface decay . Salivary levels of yeasts, which had previously been associated with decay in this population, were no longer significant using this construct . CONCLUSIONS: This construct of reporting salivary bacteriological data as a function of tooth number and per ml saliva could improve the reliability of bacteriological data obtained in epidemiological studies investigating the role of bacteria in dental decay in the elderly. Plasmid, 2000 Mar, 43(2), 130 - 6 Nucleotide sequence analysis of the lactococcal EPS plasmid pNZ4000; van Kranenburg R et al.; The complete 42180-bp nucleotide sequence of the mobilization plasmid pNZ4000, coding for exopolysaccharide (EPS) production in Lactococcus lactis, was determined . This plasmid contains a region involved in EPS biosynthesis, four functional replicons, a region containing mobilization genes, and three origin of transfer (oriT) sequences . Sequences identical to these oriT sequences were also found on two other lactococcal plasmids and a plasmid from Lactobacillus helveticus . Several complete and partial IS elements were identified on pNZ4000, including iso-ISS1, iso-IS946, and iso-IS982 sequences . Furthermore, pNZ4000 contains a gene cluster that may encode a cobalt transport system and a gene that encodes a CorA homologue which may function as a magnesium transporter . Poult Sci, 2000 Jan, 79(1), 7 - 11 Lactobacillus flora in the cloaca and vagina of hens and its inhibitory activity against Salmonella enteritidis in vitro; Miyamoto T et al.; Lactobacilli in the cloaca and vagina of 40 normal laying hens were investigated quantitatively and qualitatively, and their ability to inhibit growth of Salmonella enteritidis (SE) was examined using a spot-the-lawn technique . All samples of cloacal contents and half the samples of vaginal mucus were positive for lactobacilli . The means +/- SD of total Lactobacillus counts in the cloaca and those in the vagina were log10 5.5 +/- 1.1 and 2.5 +/- 2.6 cfu/g, respectively . In the cloaca, Lactobacillus acidophilus was isolated from 92.5% of hens, and Lactobacillus salivarius was isolated from 85.0% of hens, whereas Lactobacillus fermentum was isolated from only one hen . In the vagina, L . acidophilus and L . salivarius were isolated from 42.5% of hens . In the inhibition assay in vitro, all strains of Lactobacillus from cloacal contents and vaginal mucus inhibited growth of SE . There was a wide range of the inhibitory activity even in the same species . No difference of the growth inhibition zone was observed between lactobacilli from cloaca and those from vagina . The present study suggested that lactobacilli in the cloaca and vagina of hens might have a protective effect against SE colonization. Biochim Biophys Acta, 2000 Feb 29, 1490(3), 355 - 61 A gene cluster encoding plantaricin 1.25beta and other bacteriocin-like peptides in Lactobacillus plantarum TMW1.25; Ehrmann MA et al.; Plantaricin 1.25beta is a thermostable class two bacteriocin produced by Lactobacillus plantarum TMW1.25 isolated from sausage fermentation . It is co-produced with several other bacteriocin-like peptides . Using oligonucleotides derived from previously determined peptide sequences, a 3.8 kb DNA fragment could be amplified . A neighboring 1.8 kb fragment was amplified using ligation-anchored single-specific-primer PCR . Sequencing of the complete 5.6 kb stretch revealed that the structural gene for plantaricin 1.25beta, plnB, was located downstream of another bacteriocin gene, plnC . Seven other open reading frames were detected, including plnK encoding a bacteriocin-like peptide, but not including any putative immunity genes . Interestingly, the gene cluster contained an IS30-like insertion sequence, designated IS125, as well as an ISS1 homolog. Hua Xi Yi Ke Da Xue Xue Bao, 1998 Mar, 29(1), 33 - 7 {The effect of removable partial denture on the composition of bacteria on the apt-to-decay site of abutment teeth}; Gao N et al.; This study was aimed at the composition of plaques which were on the base teeth's carious sensitive sites, just after the insertion of the removable partial denture (RPD) . The results showed that the ratios of bacterial detection in different teeth at the same period and those on the different sites of same teeth at different periods were not significantly different . There was a tendency that the detection ratio of the stomatococcus at the early period was higher than that of other bacteria, but at the middle period the ratios declined, and at the late period it was higher again . The detection ratio of the bacillus was on the contrary in the study . At the same time, the composition of the plaques changed significantly after the dentures had been used for 7, 14, and 21 days and 3 months . After 3 months, the ratios of the stomatococcus and the bacillas returned to their normal levels . The 3 main cariogenic bacteria were all detected in this study . The Streptococcus mutant was the dominant bacterium in the plaques and its ratio went up with the time duration . The ratio for the lower second molar was higher than that for the mandibular second premolar (P < 0.05) . The detection ratios of the Actinomyces viscosus and lactobacillus were lower . In conclusion, after the insertion of RPD, the oral microbial ecosystem would be changed and the cariogenic bacteria began to implant on the surface of the base teeth . So RPD is a potential factor to cause the caries. Rinsho Biseibutshu Jinsoku Shindan Kenkyukai Shi, 1999 Dec, 10(2), 77 - 82 Identification of Lactobacillus species of human origin by a commercial kit, API50CHL; Song YL et al.; The efficacy of a biochemical kit, API50CHL kit, for identification of intestinal and vaginal lactobacilli from humans was evaluated by comparing with the results of DNA-DNA hybridization assay . The results showed that in total only 52 of the 172 strains (30.2%) tested were identified correctly by the kit at species level . Especially all strains of some species, such as Lactobacillus gasseri and Lactobacillus crispatus, were misidentified as Lactobacillus acidophilus by the kit . However, the kit was found to be reliable for identification of Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus rhamnosus, and Lactobacillus salivarius . This suggests that the exact identification of Lactobacillus isolates from human stool and vaginal specimens by API50CHL kit is difficult without the support of modern genotypic technique. Sex Transm Dis, 2000 Feb, 27(2), 74 - 8 Effect of chlorhexidine on genital microflora, Neisseria gonorrhoeae, and Trichomonas vaginalis in vitro; Rabe LK et al.; BACKGROUND: Chlorhexidine is a disinfectant that has been used in skin and mouth washes and as a preservative in some vaginal lubricants . A gel containing 0.25% chlorhexidine gluconate has been found to be effective against Chlamydia trachomatis in vitro and in animal models . Applied vaginally, 5 g of this gel could achieve vaginal fluid concentrations of < or = 1250 microg/ml . GOAL: To test the in vitro activity of chlorhexidine in a gel over a pH range of 4 to 8 in the presence or absence of blood . STUDY DESIGN: Organisms were exposed to chlorhexidine for 30 minutes to 2 hours, and the minimum cidal concentration (MCC) was calculated . RESULTS: The MCC for Neisseria gonorrhoeae was 25 microg/ml at 30 minutes and 12.5 microg/ml at 1 to 2 hours of exposure, whereas the MCC for Trichomonas vaginalis was 1250 microg/ml . Chlorhexidine was more active at pH 8 than pH 4, and less active in the presence of blood . The MCC for Lactobacillus crispatus was 1250 microg/ml at pH 4 and only 125 microg/ml at pH 8 . CONCLUSIONS: Based on its in vitro activity, chlorhexidine may be an appropriate topical microbicide for prevention of gonorrhea, but not for prevention of trichomoniasis . This study suggests that the presence of blood and pH affect the activity of chlorhexidine against genital pathogens and commensals. Zentralbl Veterinarmed B, 1999 Dec, 46(10), 683 - 7 Aggregation of sow lactobacilli with diarrhoeagenic Escherichia coli; Kmet V et al.; A total of 20 strains of lactobacilli were isolated from the oesophagus and vagina of 20 sows at the time of partus . Aggregation activity was seen between six homofermentative autoaggregative lactobacilli and three strains of pathogenic Escherichia coli with F4, F5 and F6 fimbriae . The highest aggregation activity was observed between vaginal Lactobacillus acidophilus PV 32 or oesophageal OE 2/1 and E . coli with F4 (K88) . The presence of aggregation-promoting factor (APF) was confirmed by polymerase chain reaction (PCR) with primers of a specific fragment the apf gene derived from human L . gasseri 4B2 in one oesophageal L . acidophilus strain OE 2/1 . We propose that autoaggregative lactobacilli that aggregate with diarrhoeagenic E . coli can express a class of APF proteins that exhibit the function of an aggregation mediator. J Infect Dis, 2000 Feb, 181(2), 595 - 601 Effects of contraceptive method on the vaginal microbial flora: a prospective evaluation; Gupta K et al.; A prospective evaluation of 331 university women who were initiating use of oral contraceptive pills (OCPs), a cervical cap, diaphragm-spermicide, or other spermicidal methods was done to assess the effects of commonly used contraceptive methods on the vaginal flora . Vaginal introital cultures were performed at baseline and then weekly for 1 month . The prevalence of Escherichia coli vaginal colonization and of abnormal vaginal Gram stain scores (Nugent criteria) increased significantly among women using a cervical cap or diaphragm-spermicide but not among women using OCPs . Women with E . coli colonization were significantly more likely to have an abnormal Nugent score and an absence of lactobacilli . In a multivariate model, use of spermicidal contraception in the preceding week was associated with an abnormal Nugent score and with colonization with E . coli, Enterococcus species, and anaerobic gram-negative rods . Thus, spermicidal methods of contraception are associated with alterations of the vaginal microflora that consequently may predispose women to genitourinary infections. J Infect Dis, 2000 Feb, 181(2), 587 - 94 Intravaginal practices, vaginal flora disturbances, and acquisition of sexually transmitted diseases in Zimbabwean women; van De Wijgert JH et al.; One hundred sixty-nine Zimbabwean women were studied to determine whether the use of intravaginal practices (cleaning with the fingers, wiping the vagina, and inserting traditional substances) are associated with disturbances of vaginal flora and acquisition of sexually transmitted diseases (STDs) . Subjects were interviewed and received counseling and a pelvic examination at enrollment, 1 month, and 6 months, and vaginal specimens were collected at enrollment and at 6 months . Users were more likely than nonusers to have vaginal flora disturbances but were not more likely to acquire an STD (relative risk {RR}, 2.15; P=.188) . Certain vaginal flora disturbances were associated with increased STD incidence and HIV prevalence . The absence of lactobacilli from the vaginal flora was associated with being positive for human immunodeficiency virus in baseline (odds ratio {OR}, 0.24; P=.001) and 6-month transition multivariate models (OR, 0.39; P=.025) . The presence of clue cells at baseline was associated with a higher incidence of STDs (RR, 1 . 94; P=.025). J Dent, 2000 Feb, 28(2), 111 - 6 A five-year clinical evaluation of Class II composite resin restorations; Kohler B et al.; OBJECTIVES: To study the clinical efficacy of posterior composite resin restorations placed in general practice after five years . METHODS: Two commercial composite resin systems were used . Three general practitioners placed the restorations at a Public Dental Health Service Office . The patients were not selected specifically for this study . Class II cavities were restored with Superlux Molar and P-50 APC composite systems on an alternate basis . At baseline, 63 restorations were placed in molars and premolars in 45 patients . For primary caries, generally a conservative cavity design was used (n = 23), while replacements of amalgams resulted in the larger conventional Class II design (n = 40) . The restorations were assessed using a modified USPHS criteria . Wear measurements were determined by the Leinfelder method . Photographs and bite-wing radiographs were taken to supplement the clinical evaluation of colour match, marginal adaptation and recurrent caries . Saliva sampling was performed to determine the rate of secretion and the level of mutans streptococci and lactobacilli . RESULTS: At the five-year review 51 restorations were available for examination, of which nine restorations were rated clinically unacceptable . Including the failed restorations (n = 7), at the three-year review, a total of 16 restorations had failed (27.6%) over a five-year period . The most common reasons for failure were recurrent caries (n = 7) and marginal defects (n = 4) . The mean wear of Superlux Molar was 167 microns and of P-50 APC 158 microns . Eight of the 11 patients with failed restorations due to caries and marginal defects had high counts of mutans streptococci at baseline . CONCLUSIONS: The failures in the present group of patients did not specifically relate to material, tooth type or cavity design . However, it is suggested that patient factors such as caries activity should be monitored and managed. Acta Crystallogr D Biol Crystallogr, 1999 Dec, 55 ( Pt 12), 1978 - 85 Three-dimensional structure of the Gly121Tyr dimeric form of ornithine decarboxylase from Lactobacillus 30a; Vitali J et al.; Ornithine decarboxylases catalyze the conversion of ornithine to putrescine at the beginning of the polyamine pathway . Ornithine decarboxylase (ODC) from Lactobacillus 30a is a 990612 Da dodecamer composed of six homodimers . A single point mutation (Gly121Tyr) was found to prevent association of dimers into dodecamers . The dimeric protein has been crystallized at pH 7.0 in the presence of guanosine triphosphate (GTP) . Crystals belong to space group P3(2)21, with unit-cell parameters a = 111.8, c = 135.9 A and one monomer in the asymmetric unit . The structure was determined by molecular replacement and refined using simulated annealing to R = 0.211 at 2 . 7 A resolution . The GTP-binding site was analyzed in detail . The protein exhibits a novel binding mode for GTP which is different from that seen in most G-proteins or GTPases . Central to this binding scheme appear to be three lysines, Lys190, Lys374 and Lys382, which form salt bridges with the three phosphates, and Thr191, which hydrogen bonds with the guanine base . Furthermore, the structure suggests that there is some flexibility in the wing domain, which can change its orientation as the protein adapts to its environment . The active site is similar to that of the native enzyme, consistent with the observation that the enzyme activity does not depend on its dodecameric state. Int J Clin Pract, 1999 Apr-May, 53(3), 179 - 84 The effect of supplementation with milk fermented by Lactobacillus casei (strain DN-114 001) on acute diarrhoea in children attending day care centres; Pedone CA et al.; The objective of this study was to determine if supplementation with milk fermented by yogurt cultures and Lactobacillus casei (strain DN-114 001) could lessen acute diarrhoea in healthy children . The study was conducted over six months, with 287 children aged 18.9 (SD 6.0) months, comprising three periods of one month supplementation, each month being followed by one month without supplementation . Subjects were supplemented daily with either 125 g or 250 g (according to age) of one of three tested dairy products: standard yogurt, milk fermented by yogurt cultures and Lactobacillus casei (10(8) cfu/ml), or a jellied milk (control product) . A daily record was kept of the number and type of stools . Although the incidence of diarrhoea was not shown to be different between the groups, the severity of diarrhoea over the six-month study was significantly decreased (4.3 days) with the supplementation of L . casei fermented milk compared with the jellied milk (8.0 days) (p = 0.009). Biotechnol Prog, 2000 Jan-Feb, 16(1), 59 - 63 Characterization of Lactobacillus carbohydrate fermentation activity using immobilized cell technique; Corton E et al.; A microbial bioreactor based on calcium alginate immobilized Lactobacillus cells coupled to a pH electrode was developed for quantitative determination of carbohydrate fermentation activity . A high biomass (10(10) cfu mL(-)(1)) and particular pregrowth conditions were needed . Reduction of catabolite repression by monosaccharides was achieved by pregrowth in lactose . The evolution of acid production in a continuous flow-stopped flow bioreactor was monitored for different sugar solutions in contact with the immobilized bacteria . The resulting slopes (DeltamV/Deltat) were used to quantify the fermentation capability for a defined sugar related to that of glucose, which was taken as 100% . The procedure is simple, being based on pH variation that can give quantitative results compared to other reported techniques for carbohydrate fermentation pattern from which only qualitative results are obtained . In addition, it offers reduction in time and costs and is a suitable tool for the rapid analysis of isolated strains and in studies of modifications of sugar metabolism in mutants. J Appl Microbiol, 1999 Dec, 87(6), 923 - 31 Purification and characterization of a bacteriocin produced by Lactobacillus acidophilus IBB 801; Zamfir M et al.; Lactobacillus acidophilus IBB 801 produces a small bacteriocin, designated acidophilin 801, with an estimated molecular mass of less than 6.5 kDa . It displays a narrow inhibitory spectrum (only related lactobacilli but including the Gram-negative pathogenic bacteria Escherichia coli Row and Salmonella panama 1467) with a bactericidal activity . The antimicrobial activity of cell-free culture supernatant fluid was insensitive to catalase but sensitive to proteolytic enzymes such as trypsin, proteinase K and pronase, heat-stable (30 min at 121 degrees C), and maintained in a wide pH range . The proteinaceous compound was isolated from cell-free culture supernatant fluid and purified . Crude bacteriocin was isolated as a floating pellicle after ammonium sulphate precipitation (40% saturation) and partially purified by extraction/precipitation with chloroform/methanol (2/1, v/v) . Further purification to homogeneity was performed by reversed phase Fast Performance Liquid Chromatography . The amino acid composition was determined . Amino acid sequencing revealed that the N-terminal end was blocked. Int J Gynaecol Obstet, 1999 Nov, 67 Suppl 1, S35 - 8 The future of bacterial vaginosis-related research; Taylor-Robinson D; Various ways and criteria are used to diagnose BV . Guidelines should be redrawn and they should embody greater uniformity . The etiology of BV remains enigmatic . However, various observations suggest that host factors, possibly hormonal, cause an imbalance in the vaginal microflora . Exogenous factors, such as semen and antibiotics, may then help to bring about a more prolonged change . This forms a working hypothesis for further exploration . The role of the lactobacillus phage in the development of BV also needs to be determined . Various conditions may occur as a consequence of BV in non-pregnant and pregnant women and BV may also affect men . A subjective assessment of the extent to which these associations occur or are likely to be shown to occur by further investigations is presented in Table 1 . The ability to cure acute BV needs to be improved as does the treatment of chronic BV, for which vaginal recolonization with exogenous lactobacilli is an approach to be evaluated further. Cancer Lett, 1999 Dec 1, 147(1-2), 125 - 37 Inhibitory effects of freeze-dried milk fermented by selected Lactobacillus bulgaricus strains on carcinogenesis induced by 1,2-dimethylhydrazine in rats and by diethylnitrosamine in hamsters; Balansky R et al.; Fermented milk products might be used for cancer chemoprevention due to their putative anticarcinogenic and antitumor activities . The diet was supplemented with freeze-dried milk fermented by Lactobacillus bulgaricus strain LBB.B 144 (product FFM.B 144) added throughout the experiment at doses of 1.3 g and 2.5 g per rat, 5 times a week starting 3 weeks before the first carcinogen injection . This treatment significantly inhibited, by 26.2-28.6% and by 34.2%, the total intestinal carcinogenesis induced by 1,2-dimethylhydrazine (DMH, 21 mg/kg, s.c., once per week for 20 weeks) in male and female BD6 rats, respectively . FFM.B144 decreased the tumor incidence and multiplicity in large bowel, caecum, and duodenum . Protective effects were better expressed in female animals, with exception of that observed in duodenum . Supplementation of diet with freeze-dried milk fermented by Lactobacillus bulgaricus strain LBB.B5 (product FFM.B5) inhibited DMH-induced carcinogenesis only in the large bowel, but had no significant protective effect when all intestinal tumors were taken into account . However, both freeze-dried products favorably shifted the differentiation of large bowel tumors by increasing the proportion of benign and highly differentiated malignant tumors and decreasing in parallel the number of poorly differentiated carcinomas without influencing the tumor size . A lower number of cases with visible mesenterial metastasis was also observed in FFM-treated rats . In addition, both FFM.B 144 and FFM.B5 significantly inhibited, by 26-33%, the induction in the same rats of ear-duct tumors . FFM.B144 but not FFM.B5 was also effective in inhibiting the tracheal carcinogenesis induced in Syrian golden hamsters by diethylnitrosamine (DEN, 100 mg/kg, two s.c . injections), the protective effect being better expressed in female animals . The anticarcinogenic potential of some fermented milk products might be exploited in chemoprevention of cancer in humans. Adv Exp Med Biol, 1999, 473, 185 - 90 Potentiation of the effectiveness of Lactobacillus casei in the prevention of E . coli induced diarrhea in conventional and gnotobiotic pigs; Bomba A et al.; The influence of preventive administration of Lactobacillus casei subsp . casei and maltodextrin KMS X-70 on Escherichia coli 08: K88 adhesion in the gastrointestinal tract of 11 conventional and 6 gnotobiotic piglets was investigated . The preventive administration of L . casei alone had almost no inhibitory effect on the adherence of E . coli to the jejunal mucosa of gnotobiotic and conventional piglets while the lactobacilli administered together with maltodextrin decreased the number of E . coli colonising jejunal mucosa of gnotobiotic piglets by 1 logarithm (4.95 log 10/cm2) in comparison with the control group (5.96 log 10/cm2) . L . casei administered in combination with maltodextrin decreased the number of E . coli colonising the jejunum of conventional piglets by more than two and half logarithm (4.75 log 10/cm2, p < 0.05) in comparison with the control (7.42 log 10/cm2) . The inhibitory effect of Lactobacillus casei and maltodextrin KMS X-70 on the adhesion of E . coli to the intestinal mucosa of conventional and gnotobiotic pigs was probably mediated by Lactobacillus--produced antibacterial substances and stimulation of immunity. Appl Environ Microbiol, 2000 Feb, 66(2), 606 - 13 Bacteriocin production with Lactobacillus amylovorus DCE 471 is improved and stabilized by fed-batch fermentation; Callewaert R et al.; Amylovorin L471 is a small, heat-stable, and hydrophobic bacteriocin produced by Lactobacillus amylovorus DCE 471 . The nutritional requirements for amylovorin L471 production were studied with fed-batch fermentations . A twofold increase in bacteriocin titer was obtained when substrate addition was controlled by the acidification rate of the culture, compared with the titers reached with constant substrate addition or pH-controlled batch cultures carried out under the same conditions . An interesting feature of fed-batch cultures observed under certain culture conditions (constant feed rate) is the apparent stabilization of bacteriocin activity after obtaining maximum production . Finally, a mathematical model was set up to simulate cell growth, glucose and complex nitrogen source consumption, and lactic acid and bacteriocin production kinetics . The model showed that bacterial growth was dependent on both the energy and the complex nitrogen source . Bacteriocin production was growth associated, with a simultaneous bacteriocin adsorption on the producer cells dependent on the lactic acid accumulated and hence the viability of the cells . Both bacteriocin production and adsorption were inhibited by high concentrations of the complex nitrogen source. Immunol Cell Biol, 2000 Feb, 78(1), 55 - 66 Immune response to orally consumed antigens and probiotic bacteria; Chin J et al.; The gut mucosal system must fulfil conflicting roles in suppressing immune responses against orally fed antigens (tolerance) while still retaining the ability to respond to potential enteric pathogens . It must also, to a large degree, not mount an immune response against commensal enteric bacteria and the administration of large numbers of probiotic bacteria formulated as dietary supplements in food products . Contrary to this dogma, it has been found that feeding ovalbumin as a marker antigen, in association with selected probiotic bacteria, appears to prime for an intestinal immune response that is further augmented by skin vaccination . Skin immunization is known to stimulate a strong innate, humoral and cellular immune response . Such dominant immunogenic signals appear to override tolerogenic signals engendered by oral feeding of antigen . High-dose antigen feeding stimulated a strong Th2-dependent antibody response to skin vaccination but completely suppressed cytotoxic T cell responses . This was true even when ovalbumin was administered in conjunction with various selected probiotic bacteria . However, while yeast appeared to be better at priming for an enhanced humoral response, Lactobacillus fermentum and Staphylococcus carnosus were more effective in enhancing the postvaccinal lymphoproliferative response against ovalbumin. Appl Microbiol Biotechnol, 1999 Dec, 53(1), 69 - 74 Isolation and characterisation of a ropy Lactobacillus strain producing the exopolysaccharide kefiran; Micheli L et al.; A capsular-polysaccharide-producing strain, LM-17, was isolated from kefir grains and was identified as a slime-forming, rod-shaped Lactobacillus . According to 1H- and 13C-NMR spectral data, the exopolysaccharide produced by the isolated bacterial strain is identical to the glucogalactan extracted from kefir grains and therefore known as kefiran . The kefiran produced was characterised by means of viscosity, optical rotatory power, circular dichroism and IR spectral measurements . A batch procedure was set up for the culture and extraction of the exopolysaccharide in laboratory conditions, resulting in a yield of 2 g/l purified kefiran from the culture supernatant of the LM-17 strain. J Inorg Biochem, 1999 Nov-Dec, 77(3-4), 185 - 95 Thermolysis of coenzymes B12 at physiological temperatures: activation parameters for cobalt-carbon bond homolysis and a quantitative analysis of the perturbation of the homolysis equilibrium by the ribonucleoside triphosphate reductase from Lactobacillus leichmannii; Brown KL et al.; The kinetics of the thermolysis of 5'-deoxyadenosylcobalamin (AdoCbl, coenzyme B12) in aqueous solution, pH 7.5, have been studied in the temperature range 30-85 degrees C using AdoCbl tritiated at the adenine C2 position and the method of initial rates . Combined with a careful analysis of the distribution of adenine-containing products, the results permit the dissection of the competing rate constants for carbon-cobalt bond homolysis and heterolysis . After correction for the temperature-dependent occurrence of the much less reactive base-off species of AdoCbl, the activation parameters for homolysis of the base-on species were found to be delta H++homo,on = 33.8 +/- 0.2 kcal mol-1 and delta S++homo,on = 13.5 +/- 0.7 cal mol-1 K-1, values not significantly different from those determined by Hay and Finke (J . Am . Chem . Soc . 108 (1986) 4820), in the temperature range 85-115 degrees C . In contrast, the heterolysis of base-on AdoCbl was characterized by a much smaller enthalpy of activation (delta H++het,on = 18.5 +/- 0.2 kcal mol-1) and a negative entropy of activation (delta S++het,on = -34.0 +/- 0.7 cal mol-1 K-1) so that heterolysis, which is minor pathway at elevated temperatures, is the dominant pathway for AdoCbl decomposition at physiological temperatures . Using literature values for the rate constant for the reverse reaction, the equilibrium constant for AdoCbl homolysis at 37 degrees C was calculated to be 7.9 x 10(-18) . Comparison with the equilibrium constant for this homolysis at the active site of the ribonucleoside triphosphate reductase from Lactobacillus leichmannii shows that the enzymes shifts the equilibrium constant towards homolysis products by a factor of 2.9 x 10(12) (17.7 kcal mol-1) by binding the thermolysis products with an equilibrium constant of 7.1 x 10(16) M-2, compared to the bonding constant for AdoCbl of 2.4 x 10(4) M-1. Eur J Cancer Prev, 1999 Dec, 8(6), 533 - 7 Effects of intestinal bacteria on the development of colonic neoplasm II . Changes in the immunological environment; Horie H et al.; To study the effects of intestinal bacteria on the development of colonic neoplasm, we have established gnotobiotic mice with a single species of intestinal bacteria . In the previous study, the incidence of colonic adenoma induced with 1,2-dimethylhydrazine (DMH) in the gnotobiotic mice with Lactobacillus acidophilus, gnotobiotic mice with Escherichia coli and germ-free mice were 30, 50 and 74%, respectively . In this study, 7-week-old mice in each group were sacrificed without the administration of DMH to examine the constituents of immuno-competent cells in various mouse organs using flow cytometry . In the gnotobiotic mice, CD3 intermediate interleukin (IL)-2Rbeta positive cells were observed predominantly in the liver . In the gnotobiotic mice with L . acidophilus, Mac-1 positive Gr-1 positive cells were observed predominantly in the colonic lamina propria . The activation of extrathymic T cells in the liver and granulocytes in the colonic mucosa may be related to anti-neoplastic effects of L . acidophilus in this experimental model. Microbios, 1999, 100(397), 135 - 44 Prevalence and persistence of amoxycillin-resistant bacteria in the dental plaques of adults; Packer S et al.; The prevalence and persistence of amoxycillin-resistant organisms (ARO) in the dental plaque of adults was determined . Plaque samples from ten adults, who had not taken antibiotics during the previous 6 months, were screened for ARO on three occasions at intervals of 3 months . The ARO were tested for their susceptibility to amoxycillin and to amoxycillin plus clavulanic acid as well as their ability to produce beta-lactamases . The ARO were found in all subjects on at least one sampling occasion and in 87% of the 30 samples examined . Of the 36 ARO isolated, 33% were yeasts, 19% were staphylococci, 19% Actinomycetes spp . and 14% lactobacilli, whilst seventeen of the isolates produced a beta-lactamase and seven of these were sensitive to coamoxiclav . The proportion of ARO in an individual fluctuated widely over the study period . It is suggested that the ARO are frequently, though transiently, present in low numbers in the plaque of individuals who have not recently received antibiotics. Infect Immun, 2000 Feb, 68(2), 752 - 9 Activation of human peripheral blood mononuclear cells by nonpathogenic bacteria in vitro: evidence of NK cells as primary targets; Haller D et al.; The interaction of commensal bacteria with immunocompetent cells may occur in definite compartments of the mucosal immune system, as limited translocation through the epithelial barrier cannot be excluded . In this study the stimulation of human peripheral blood mononuclear cells and purified lymphocyte subsets by nonpathogenic gram-positive lactobacilli (Lactobacillus johnsonii and Lactobacillus sakei) and gram-negative Escherichia coli was investigated . The various bacterial strains induced a differential cytokine pattern . Whereas L . johnsonii and L . sakei strongly induced gamma interferon (IFN-gamma) and interleukin-12 (IL-12), E . coli and lipopolysaccharide (LPS) preferentially induced IL-10 after 16 h of stimulation . Expression of activation antigens CD69 and CD25 was observed on (CD3(-) CD56(+)) natural killer (NK) cells after stimulation of total human peripheral blood mononuclear cells . All bacteria mediated the proliferation of human peripheral blood mononuclear cells, and the strongest proliferative response was observed with L . johnsonii . Purified CD4(+), CD8(+), and CD19(+) lymphocyte subsets were not activated upon bacterial stimulation but showed normal response to a mitogenic stimulus . In contrast, purified NK cells upregulated the IL-2Ralpha chain (CD25) and underwent proliferation when stimulated by L . johnsonii . E . coli and LPS were less effective in inducing proliferation . Expression of CD25 or secretion of IFN-gamma from purified NK cells was significantly increased in the presence of bacterially primed macrophages, indicating that full activation required both bacterium- and cell contact-based signals derived from accessory cells. Int J Food Microbiol, 1999 Dec 15, 53(2-3), 115 - 25 Hydrolysis of muscle myofibrillar proteins by Lactobacillus curvatus and Lactobacillus sake; Sanz Y et al.; Proteolytic enzyme activities of whole cells and cell free extracts (CFE) of Lactobacillus curvatus CECT 904 and Lactobacillus sake CECT 4808 were characterised using synthetic chromogenic compounds and myofibrillar proteins as substrates . The hydrolytic action was monitored by SDS-PAGE and reverse phase-HPLC analyses . The CFE of L . sake partially contributed, together with muscle enzymes, to the initial hydrolysis of myofibrillar proteins . Whole-cells of both L . curvatus and L . sake generated peptides considered important for cured-meat taste . The peptide mapping, resulting from the action on the substrates assayed, revealed a profile of extra and intracellular enzymes . Both strains expressed strong amino acid metabolism. Am J Gastroenterol, 2000 Jan, 95(1 Suppl), S22 - 5 Probiotics and immune response; Cunningham-Rundles S et al.; Current evidence supports the concept that oral administration of probiotic lactobacilli may be therapeutic in preventing antibiotic-associated diarrhea in children and in reestablishing normal flora in the gastrointestinal tract . Children with human immunodeficiency virus (HIV) infections may have episodes of diarrhea and frequently experience malabsorption associated with possible bacterial overgrowth; together these may interact to produce the growth abnormalities characteristic of this group . The overall objective of this investigation has been to determine whether oral administration of the probiotic Lactobacillus plantarum 299v could improve nutrient status and promote growth in children congenitally exposed to HIV . In addition, the possible beneficial effect of Lactobacillus plantarum 299v in modulating immune response was evaluated . In preliminary results described here, we report on the ability of Lactobacillus plantarum 299v to colonize children with HIV and to elicit specific systemic immune response after oral supplementation. Am J Gastroenterol, 2000 Jan, 95(1 Suppl), S19 - 21 Probiotics and inflammatory bowel diseases; Schultz M et al.; The pathogenesis of inflammatory bowel diseases remains elusive . However, the resident luminal bacteria seem to be an important factor in their development and chronicity . There is evidence to suggest that inflammatory bowel diseases may represent an aggressive immunological response to the resident luminal flora, rather than an alteration in the normal flora . In prior research, probiotic bacteria were effective in managing certain acute diarrheal diseases, and investigators reported that certain Lactobacilli strains seem to have protective immunomodulating and bowel flora manipulating properties . We report the results of recent studies with probiotics in animal models, in which promising effects for the treatment of chronic inflammatory bowel disease, pouchitis, and ulcerative colitis were observed . Future research may clarify a precise role for probiotic bacteria in managing chronic inflammatory bowel diseases. Am J Gastroenterol, 2000 Jan, 95(1 Suppl), S8 - 10 The effects of antibiotic use on gastrointestinal function; Levy J; The bacterial flora of the gastrointestinal (GI) tract plays an important role in maintaining the integrity of the enterocyte, modulating metabolic and immunologic processes, and protecting against colonization by invasive pathogens . Disruption of this finely tuned and stable gut flora by antibiotics, infection, chemotherapy, or radiation has profound effects on the protective barrier and results in overgrowth by pathogens, invasion and translocation of toxins, and life-threatening infections . Use of antibiotics promotes the emergence of resistant organisms, and multiple-antibiotic resistance has become a major public health issue . Preservation of protective species or recolonization with nonpathogenic yeasts or lactobacilli during periods of stress (infections, drugs) has begun to show promise in the management of patients receiving multiple antibiotics, particularly in hospital-acquired infections. J Pediatr Gastroenterol Nutr, 2000 Jan, 30(1), 68 - 72 Clinical evaluation of the addition of lyophilized, heat-killed Lactobacillus acidophilus LB to oral rehydration therapy in the treatment of acute diarrhea in children; Simakachorn N et al.; BACKGROUND: Addition of a medication to the World Health Organization protocol for treatment of acute diarrhea in children is controversial . In this trial, the clinical efficacy of a medication (Lacteol Fort sachets; Laboratoire du Lacteol du Docteur Boucard, Houdan France) containing lyophilized heat-killed Lactobacillus acidophilus LB was assessed as an adjunct to oral rehydration therapy . METHODS: Children aged 3 to 24 months with acute diarrhea and mild or moderate dehydration were enrolled in the study . Children received oral rehydration therapy for the first 4 hours . After this first rehydration phase, undiluted milk formula or breast milk was fed alternately with oral rehydration solution . Children were fed rice gruel as tolerated . They received either one sachet containing 10 billion of lyophilized heat-killed L . acidophilus LB or placebo at admission and at 12-hour intervals for five doses . RESULTS: Seventy-three children (37 L . acidophilus LB, 36 placebo) were enrolled, of whom 40 (17 L . acidophilus LB, 23 placebo) received an antibiotic before inclusion . Rotavirus was identified in approximately 50% of the children in each group . After 24 hours of treatment, the number of rotavirus-positive children with watery stools was significantly lower (p = 0.012) in the L . acidophilus LB group . Mean duration of diarrhea was decreased (p = 0.034) with L . acidophilus LB (43.4 hours) versus placebo (57.0 hours) . This decreased duration was particularly marked in children with no antibiotic therapy before inclusion (31.1 hours): 42.9 hours for the L . acidophilus LB group versus 74.0 hours for the placebo group (p = 0.016) . CONCLUSIONS: Addition of L . acidophilus LB to oral rehydration therapy was effective in the treatment of children with acute diarrhea by decreasing the duration of diarrhea. J Pediatr Gastroenterol Nutr, 2000 Jan, 30(1), 54 - 60 Lactobacillus GG administered in oral rehydration solution to children with acute diarrhea: a multicenter European trial; Guandalini S et al.; BACKGROUND: The probiotic Lactobacillus GG is effective in promoting a more rapid recovery of acute, watery diarrhea in children with rotavirus enteritis . Very limited information is available, however, on the potential role of such agents in non-rotaviral diarrheal episodes . Furthermore, no evidence is available concerning the efficacy of Lactobacillus GG administered in the oral rehydration solution during oral rehydration therapy . A multicenter trial was conducted to evaluate the efficacy of Lactobacillus GG administered in the oral rehydration solution to patients with acute-onset diarrhea of all causes . METHODS: Children 1 month to 3 years of age with acute-onset diarrhea were enrolled in a double-blind, placebo-controlled investigation . Patients were randomly allocated to group A, receiving oral rehydration solution plus placebo, or group B, receiving the same preparation but with a live preparation of Lactobacillus GG (at least 10(10) CFU/250 ml) . After rehydration in the first 4 to 6 hours, patients were offered their usual feedings plus free access to the same solution until diarrhea stopped . RESULTS: One hundred forty children were enrolled in group A, and 147 in group B . There were no differences at admission between the groups in age, sex, previous types of feeding, previous duration of diarrhea, use of antibiotics, weight, height, weight-height percentile, prevalence of fever, overall status, degree of dehydration, and percentage of in- versus outpatients . Duration of diarrhea after enrollment was 71.9 +/- 35.8 hours in group A versus 58.3 +/- 27.6 hours in group B (mean +/- SD; P = 0.03) . In rotavirus-positive children, diarrhea lasted 76.6 +/- 41.6 hours in group A versus 56.2 +/- 16.9 hours in groups B (P < 0.008) . Diarrhea lasted longer than 7 days in 10.7% of group A versus 2.7% of group B patients (P < 0.01) . Hospital stays were significantly shorter in group B than in group A . CONCLUSIONS: Administering oral rehydration solution containing Lactobacillus GG to children with acute diarrhea is safe and results in shorter duration of diarrhea, less chance of a protracted course, and faster discharge from the hospital. J Dairy Sci, 1999 Dec, 82(12), 2536 - 42 Viability of Lactobacillus gasseri and its cholesterol-binding and antimutagenic activities during subsequent refrigerated storage in nonfermented milk; Usman et al.; The effect storage at 4 degrees C on the viability of Lactobacillus gasseri and its sodium taurocholate-deconjugating and cholesterol-binding abilities as well as desmutagenic activity was investigated . Unfermented milks containing L . gasseri strains SBT0274 and SBT0270 at 10(9) cfu/ml were prepared using 10% skim milk . Total and bile-tolerant lactobacilli for strains SBT0274 and SBT0270 generally decreased after 14 d of storage at 4 degrees C; however, viable cells of these strains were still at 10(8) cfu/ml after 28 d of storage . The amounts of cholic acid released and of cholesterol bound by strains SBT0274 and SBT0270 declined over time, especially at 21 d of storage . Antimutagenic activity of unfermented milk made from both strains was attributed to the bacterial cells, and the activity was stable during storage at 4 degrees C for 28 d. J Dairy Sci, 1999 Dec, 82(12), 2525 - 9 New binding assay and preparative trial of cell-surface lectin from Lactobacillus acidophilus group lactic acid bacteria; Matsumura A et al.; To select Lactobacillus acidophilus group bacteria as a probiotic yogurt starter, we designed a new screening method that measures the binding activity of surface layer protein to rat colonic mucin, which contains sugar chains similar to those in human colonic mucin . The B1 subgroup (Lactobacillus gasseri), which is the dominant strain in the human intestinal tract, showed the highest binding activity to rat colonic mucin among all the subgroups of L . acidophilus . The binding activity of the surface layer protein was also shown to be significantly reduced after periodate oxidation of the rat colonic mucin . This new screening method is useful for rapid selection of L . acidophilus strains that have high adhesion to the human intestinal tract . Lectin-like proteins that were bound to rat colonic mucin were isolated from the surface layer proteins with a rat colonic mucin-coated membrane and were analyzed by SDS-PAGE . A few main bands together with several minor bands were observed on the electrophoretograms obtained from the strains tested . It is possible that those lectin-like proteins contribute to adhesion of the bacterial cell to human colonic mucosa by binding specifically to carbohydrate portions. Int J Food Sci Nutr, 1999 May, 50(3), 203 - 11 Processing of quinoa (Chenopodium quinoa, Willd): effects on in vitro iron availability and phytate hydrolysis; Valencia S et al.; The effect of different processing techniques was studied on in vitro iron availability and phytate hydrolysis in high and low saponin content quinoa (Chenopodium quinoa, Willd) seeds . Water slurries of ungerminated and germinated quinoa flour were processed by cooking, soaking, and fermentation using Lactobacillus plantarum as starter . Iron solubility under physiological conditions (in vitro) was measured and used as an estimation of iron availability . Phytate (inositol hexaphosphate/IP6) and its degradation products were analysed by an HPLC method . The IP6 + IP5 content was reduced by cooking with 4 to 8%, germination with 35 to 39%, soaking with 61 to 76% and by fermentation with 82 to 98% . The highest reduction, about 98%, was obtained after fermentation of the germinated flour . Cooking had no effect on the amount of soluble iron . Iron solubility increased, however, two to four times after soaking and germination, three to five times after fermentation and five to eight times after fermentation of the germinated flour samples and was highly correlated to the reduction of IP6 + IP5 (P < 0.001) . There was no difference between the quinoa varieties with regard to phytate reduction and iron solubility . The pH in fermented samples was reduced from 6.5 to about 3.5, due to lactic acid formation. Int J Food Sci Nutr, 1999 May, 50(3), 159 - 64 Formation of biogenic amine in mayonnaise, herring and tuna fish salad by lactobacilli; Leuschner RG et al.; The effect of amino acid decarboxylase-positive lactobacilli in mayonnaise, herring and tuna fish salads on formation of biogenic amines (BA) was investigated . Commercial mayonnaise was inoculated with either of five amine-forming lactobacilli which were selected as model contaminants: Lactobacillus curvatus LTH 975 and LTH 1859 (cadaverine, putrescine, tyramine and phenylethylamine producing), L . delbrueckii LTH 1260 (tyramine and phenylethylamine forming) and L . buchneri LTH 1388 and LTH 661 (histamine forming) . Low concentrations of tyramine (4.5 ppm) were detected and an addition of precursor amino acids resulted in an increase of amine concentrations to 40 ppm putrescine, 16.5 ppm tyramine and 5.5 ppm cadaverine . Herring and tuna fish salads were inoculated either with L . curvatus LTH 975 or L . Buchneri LTH 1388 . In tuna fish salad 1 ppm putrescine, 3 ppm cadaverine, 7 ppm histamine and 28 ppm tyramine were found after 4 days when L . curvatus was added . In the corresponding herring salad putrescine (14 ppm), cadaverine (11.5 ppm), histamine (17 ppm) and tyramine (72 ppm) were detected . Fish salads containing L . buchneri displayed histamine concentrations of 900 ppm in tuna and 670 ppm in herring salad, respectively . Eight lactic acid bacteria and five yeasts, isolates from spoiled delicatessen salads and ingredients, were not able to form putrescine, cadaverine, histamine, tyramine or phenylethylamine. Biochemistry, 1999 Nov 30, 38(48), 15962 - 9 Characterization of rates of ring-flipping in trimethoprim in its ternary complexes with Lactobacillus casei dihydrofolate reductase and coenzyme analogues; Polshakov VI et al.; NMR measurements have been used to investigate rates of ring-flipping and the activation parameters for the trimethoxybenzyl ring of the antibacterial drug trimethoprim (TMP) bound to Lactobacillus casei dihydrofolate reductase (DHFR) for a series of ternary complexes formed with analogues of the coenzyme NADPH . Rates were obtained at several temperatures from line shape analyses ((13)C-edited HSQC (1)H spectra) and transfer of magnetization measurements (zz-HSQC) on complexes containing 3'-O-{(13)C}trimethoprim . Examination of the structures of the complexes indicates that ring-flipping can only be achieved following major conformational changes and transient fluctuations of the protein and coenzyme structure around the trimethoxybenzyl ring . There is no simple correlation between rates of ring-flipping and binding constants . The presence of the coenzyme nicotinamide ring (in either its reduced or its oxidized forms) in the binding site close to the trimethoxybenzyl ring moiety is the major factor reducing the ring-flipping on coenzyme binding . Thus, the ternary complex with NADPH shows the largest reduction in the rate of ring-flipping (11 +/- 3 s(-)(1) at 298 K) as compared with the binary complex (793 +/- 80 s(-)(1) at 298 K) . Complexes with NADPH analogues that either have no nicotinamide ring or are known to have their nicotinamide rings removed from the binding site show the smallest reductions . For the DHFR.TMP.NADP(+) complex where there are two conformations present, very different rates of ring-flipping were observed for the two forms . The activation parameters (DeltaH() and DeltaS()) for the ring-flipping in all the complexes are discussed in terms of the protein-ligand interactions and the possible constraints on the pathway through the transition state. Appl Environ Microbiol, 2000 Jan, 66(1), 383 - 91 Use of green fluorescent protein to tag lactic acid bacterium strains under development as live vaccine vectors; Geoffroy MC et al.; The lactic acid bacteria (LAB) are safe microorganisms which are mainly used for the preparation of fermented foods and for probiotic applications . The potential of LAB as live vehicles for the production and delivery of therapeutic molecules such as antigens is also being actively investigated today . However, very little is known about the fate of live LAB when administered in vivo and about the interaction of these microorganisms with the nasal or gastrointestinal ecosystem . For future applications, it is essential to be able to discriminate the biotherapeutic strain from the endogenous microflora and to unravel the mechanisms underlying the postulated health-beneficial effect . We therefore started to investigate both aspects in a mouse model with two LAB species presently under development as live vaccine vectors, i.e., Lactococcus lactis and Lactobacillus plantarum . We have constructed different expression vectors carrying the gfp (green fluorescent protein {GFP}) gene from the jellyfish Aequoria victoria, and we found that this visible marker was best expressed when placed under the control of the inducible strong nisA promoter from L . lactis . Notably, a threshold amount of GFP was necessary to obtain a bright fluorescent phenotype . We further demonstrated that fluorescent L . plantarum NCIMB8826 can be enumerated and sorted by flow cytometry . Moreover, tagging of this strain with GFP allowed us to visualize its phagocytosis by macrophages in vitro and ex vivo and to trace it in the gastrointestinal tract of mice upon oral administration. Appl Environ Microbiol, 2000 Jan, 66(1), 310 - 9 An explosive antisense RNA strategy for inhibition of a lactococcal bacteriophage; Walker SA et al.; The coding regions of six putative open reading frames (ORFs) identified near the phage phi31 late promoter and the right cohesive end (cos) of lactococcal bacteriophage phi31 were used to develop antisense constructs to inhibit the proliferation of phage phi31 . Two middle-expressed ORFs (ORF 1 and ORF 2) and four late-expressed ORFs (ORF 3 through ORF 6) were cloned individually between the strong Lactobacillus P6 promoter and the T7 terminator (T(T7)) to yield a series of antisense RNA transcripts . When expressed on a high-copy-number vector from a strong promoter, the constructs had no effect on the efficiency of plaquing (EOP) or the plaque size of phage phi31 . To increase the ratio of antisense RNA to the targeted sense mRNA appearing during a phage infection, the antisense cassettes containing the late-expressed ORFs (ORF 3 through ORF 6) were subcloned to pTRK360, a low-copy-number vector containing the phage phi31 origin of replication, ori31 . ori31 allows for explosive amplification of the low-copy-number vector upon phage infection, thereby increasing levels of antisense RNA transcripts later in the lytic cycle . In addition, the presence of ori31 also lowers the burst size of phage phi31 fourfold, resulting in fewer sense, target mRNAs being expressed from the phage genome . The combination of ori31 and P6::anti-ORF 4H::T(T7) resulted in a threefold decrease in the EOP of phage phi31 (EOP = 0.11 +/- 0.03 {mean +/- standard deviation}) compared to the presence of ori31 alone (EOP = 0.36) . One-step growth curves showed that expression of anti-ORF 4H RNA decreased the percentage of successful centers of infection (75 to 80% for ori31 compared to 35 to 45% for ori31 plus anti-ORF 4H), with no further reduction in burst size . Growth curves performed in the presence of varying levels of phage phi31 showed that ori31 plus anti-ORF 4H offered significant protection to Lactococcus lactis, even at multiplicities of infection of 0.01 and 0.1 . These results illustrate a successful application of an antisense strategy to inhibit phage replication in the wake of recent unsuccessful reports. Appl Environ Microbiol, 2000 Jan, 66(1), 297 - 303 Detection and identification of gastrointestinal Lactobacillus species by using denaturing gradient gel electrophoresis and species-specific PCR primers; Walter J et al.; Denaturing gradient gel electrophoresis (DGGE) of DNA fragments obtained by PCR amplification of the V2-V3 region of the 16S rRNA gene was used to detect the presence of Lactobacillus species in the stomach contents of mice . Lactobacillus isolates cultured from human and porcine gastrointesti