Dig Liver Dis, 2002 Sep, 34 Suppl 2, S81 - 3 Probiotics and Helicobacter pylori eradication; Canducci F et al.; The need for new strategies for Helicobacter pylori eradication, alternative or complementary to antibiotic therapy, has recently claimed the attention of many investigators . Pre-clinical studies have shown the inhibition of Helicobacter pylori growth by Lactobacilli and the anti-Helicobacter pylori action of Lactobacillus salivarius, Lactobacillus acidophilus and Lactobacillus casei subspecies rhamnosus strains, possibly due to the production of lactic acid or to the secretion of an autolysin . Clinical studies have demonstrated a persistent reduction in delta over baseline values at the 13C urea breath test independently of omeprazole administration with Lactobacillus acidophilus La1, the eradication in 6 out of 14 patients with Lactobacillus acidophilus alone, positive results in patients in which a standard Helicobacter pylori triple therapy was randomly supplemented with Lactobacillus acidophilus. Dig Liver Dis, 2002 Sep, 34 Suppl 2, S66 - 7 Probiotics and Crohn's disease; Prantera C et al.; Antibiotics are often employed in the treatment of Crohn's disease, with interesting results . However indiscriminate suppression of intestinal bacteria may be harmful and long-term use of antibiotics is burdened by side-effects and by the risk of developing bacterial resistance . Manipulation of enteric flora with probiotic compounds would be a possible and appealing alternative . First aim of our study has been to investigate the efficacy of this probiotic in reducing the endoscopic recurrence rate or in reducing the severity of recurrent lesions at 1 year after surgery . Secondary goal has been the reduction of the clinical recurrence rate . Forty-five patients have been randomised to receive Lactobacillus rhamnosus strain GG or placebo for 12 months . The results have shown no differences in endoscopic and clinical remission between the two groups. Dig Liver Dis, 2002 Sep, 34 Suppl 2, S63 - 5 Use of Lactobacillus-GG in paediatric Crohn's disease; Guandalini S; The potential role of luminal bacteria in initiating the abnormal immune response seen in inflammatory bowel disease is stressed by many observations . A defect in mucosal barrier function could allow luminal bacterial antigens to initiate the chronic relapsing inflammation in Crohn's disease . The potential role of luminal bacteria in initiating the abnormal immune response seen in inflammatory bowel disease is stressed by many observations . A pilot study to investigate the possible effect of Lactobacillus GG in children with active Crohn's disease was conducted . Four male patients were enrolled, median age 14.5 years (range 10-18) . In terms of clinical outcome, the patients showed significant improvement . In three patients on Lactobacillus GG, it was possible to taper the dose of steroids . Thus, although our data are obviously very preliminary, Lactobacillus GG appears to be effective in improving the clinical status of children with Crohn's disease . A multicentre study is currently being carried out in 7 US University centres in a randomized, double-blind, placebo-controlled fashion to establish the efficacy of this probiotic in children with Crohn's disease. Dig Liver Dis, 2002 Sep, 34 Suppl 2, S2 - 7 Probiotics in the third millennium; Gorbach SL; Probiotics are "living microorganisms which upon ingestion in certain numbers exert health benefits beyond inherent general nutrition" . Since 1987, when the first publication on the properties of the Lactobacillus GG was done, overall, there have been over 200 publications in peer-reviewed scientific journals . This paper will report the status and the prospectus of probiotics research at the beginning of the Third Millennium . Probiotics have proven benefits in treatment and prevention of rotavirus diarrhoea in children and reduction of antibiotic-associated intestinal side-effects . Interesting results have recently been published regarding food allergies and atopic eczema in children . Prevention of vaginitis and of travellers' diarrhoea have also been reported . Promising results are being reported in patients with inflammatory bowel disease, cystic fibrosis, dental caries and irritable bowel syndrome . It has also been suggested that probiotics could enhance oral vaccine administration, and that they may help treatment against Helicobacter pylori infection, but further studies are needed . Future areas of research regard probiotics' role in the process of carcinogenesis, given their influence on the gut microflora, and as immune modulators in autoimmune disorders . The possibility of introducing appropriate genes to the probiotics to make them produce various compounds is also under investigation . However, there is still confusion in the minds of the authorities over whether a probiotic is a drug, a food, or a dietary supplement . The challenge is to continue research to define the appropriate uses of probiotics and discover new applications which will bring benefit to humankind. Appl Environ Microbiol, 2002 Nov, 68(11), 5663 - 70 Use of the alr gene as a food-grade selection marker in lactic acid bacteria; Bron PA et al.; Both Lactococcus lactis and Lactobacillus plantarum contain a single alr gene, encoding an alanine racemase (EC 5.1.1.1), which catalyzes the interconversion of D-alanine and L-alanine . The alr genes of these lactic acid bacteria were investigated for their application as food-grade selection markers in a heterologous complementation approach . Since isogenic mutants of both species carrying an alr deletion (Deltaalr) showed auxotrophy for D-alanine, plasmids carrying a heterologous alr were constructed and could be selected, since they complemented D-alanine auxotrophy in the L . plantarum Deltaalr and L . lactis Deltaalr strains . Selection was found to be highly stringent, and plasmids were stably maintained over 200 generations of culturing . Moreover, the plasmids carrying the heterologous alr genes could be stably maintained in wild-type strains of L . plantarum and L . lactis by selection for resistance to D-cycloserine, a competitive inhibitor of Alr (600 and 200 micro g/ml, respectively) . In addition, a plasmid carrying the L . plantarum alr gene under control of the regulated nisA promoter was constructed to demonstrate that D-cycloserine resistance of L . lactis is linearly correlated to the alr expression level . Finally, the L . lactis alr gene controlled by the nisA promoter, together with the nisin-regulatory genes nisRK, were integrated into the chromosome of L . plantarum Deltaalr . The resulting strain could grow in the absence of D-alanine only when expression of the alr gene was induced with nisin. Appl Environ Microbiol, 2002 Nov, 68(11), 5607 - 19 Antibacterial activities of nisin Z encapsulated in liposomes or produced in situ by mixed culture during cheddar cheese ripening; Benech RO et al.; This study investigated both the activity of nisin Z, either encapsulated in liposomes or produced in situ by a mixed starter, against Listeria innocua, Lactococcus spp., and Lactobacillus casei subsp . casei and the distribution of nisin Z in a Cheddar cheese matrix . Nisin Z molecules were visualized using gold-labeled anti-nisin Z monoclonal antibodies and transmission electron microscopy (immune-TEM) . Experimental Cheddar cheeses were made using a nisinogenic mixed starter culture, containing Lactococcus lactis subsp . lactis biovar diacetylactis UL 719 as the nisin producer and two nisin-tolerant lactococcal strains and L . casei subsp . casei as secondary flora, and ripened at 7 degrees C for 6 months . In some trials, L . innocua was added to cheese milk at 10(5) to 10(6) CFU/ml . In 6-month-old cheeses, 90% of the initial activity of encapsulated nisin (280 +/- 14 IU/g) was recovered, in contrast to only 12% for initial nisin activity produced in situ by the nisinogenic starter (300 +/- 15 IU/g) . During ripening, immune-TEM observations showed that encapsulated nisin was located mainly at the fat/casein interface and/or embedded in whey pockets while nisin produced by biovar diacetylactis UL 719 was uniformly distributed in the fresh cheese matrix but concentrated in the fat area as the cheeses aged . Cell membrane in lactococci appeared to be the main nisin target, while in L . casei subsp . casei and L . innocua, nisin was more commonly observed in the cytoplasm . Cell wall disruption and digestion and lysis vesicle formation were common observations among strains exposed to nisin . Immune-TEM observations suggest several modes of action for nisin Z, which may be genus and/or species specific and may include intracellular target-specific activity . It was concluded that nisin-containing liposomes can provide a powerful tool to improve nisin stability and availability in the cheese matrix. Appl Environ Microbiol, 2002 Nov, 68(11), 5429 - 36 Food-grade delivery system for controlled gene expression in Lactococcus lactis; Henrich B et al.; A food-grade system for the delivery of desired genes to Lactococcus lactis, their inducible expression, and their transfer to related strains was established . Based on the thermosensitive pG(+)host replicon, two types of plasmid vectors were constructed which contained sections of either the chromosomal leu operon of L . lactis or the tel operon from the lactococcal sex factor . Genes cloned into the leu or tel sequences of these vectors were delivered to the homologous regions of the chromosome or the sex factor through two single crossovers, leading to integration of the recombinant plasmids and subsequent excision of the vector portions . Inducible transcription of integrated genes was achieved by using the nisin-controlled expression (NICE) system . To establish the signal transduction genes nisRK in L . lactis, the vectors pLNG1363 (targeted to the chromosome) and pUK500 (targeted to the sex factor) were constructed . Fusions of six different peptidase genes (pep) from Lactobacillus delbrueckii with the nisin-inducible promoter P(nisA) were delivered to the sex factor with derivatives of the vector pUK300 . Food-grade recombinants of L . lactis were constructed which had the nisRK genes and individual P(nisA)::pep fusions integrated either separately into the chromosome and the sex factor or simultaneously into the sex factor . With both types of recombinants, expression of P(nisA)::pep fusions after induction with nisin was demonstrated . Depending on the loci used for integration of nisRK, variable induction rates were observed . Furthermore, an engineered sex factor carrying a P(nisA)::pepI fusion was transfered by conjugation between two strains of L . lactis at a frequency of 4 x 10(-4). Appl Environ Microbiol, 2002 Nov, 68(11), 5379 - 86 Evaluation of the heat inactivation of Escherichia coli and Lactobacillus plantarum by differential scanning calorimetry; Lee J et al.; Differential scanning calorimetry (DSC) is used to evaluate the thermal stability and reversibility after heat treatment of transitions associated with various cellular components of Escherichia coli and Lactobacillus plantarum . The reversibility and the change in the thermal stability of individual transitions are evaluated by a second temperature scan after preheating in the DSC to various temperatures between 40 and 130 degrees C . The viability of bacteria after a heat treatment between 55 and 70 degrees C in the DSC is determined by both plate count and calorimetric data . The fractional viability values based on calorimetric and plate count data show a linear relationship . Viability loss and the irreversible change in DSC thermograms of pretreated whole cells are highly correlated between 55 and 70 degrees C . Comparison of DSC scans for isolated ribosomes shows that the thermal stability of E . coli ribosomes is greater than that of L . plantarum ribosomes, consistent with the greater thermal tolerance of E . coli observed from viability loss and DSC scans of whole cells. Appl Environ Microbiol, 2002 Nov, 68(11), 5374 - 8 Membrane-bound ATPase contributes to hop resistance of Lactobacillus brevis; Sakamoto K et al.; The activity of the membrane-bound H+-ATPase of the beer spoilage bacterium Lactobacillus brevis ABBC45 increased upon adaptation to bacteriostatic hop compounds . The ATPase activity was optimal around pH 5.6 and increased up to fourfold when L . brevis was exposed to 666 microM hop compounds . The extent of activation depended on the concentration of hop compounds and was maximal at the highest concentration tested . The ATPase activity was strongly inhibited by N,N'-dicyclohexylcarbodiimide, a known inhibitor of FoF1-ATPase . Western blots of membrane proteins of L . brevis with antisera raised against the alpha- and beta-subunits of FoF1-ATPase from Enterococcus hirae showed that there was increased expression of the ATPase after hop adaptation . The expression levels, as well as the ATPase activity, decreased to the initial nonadapted levels when the hop-adapted cells were cultured further without hop compounds . These observations strongly indicate that proton pumping by the membrane-bound ATPase contributes considerably to the resistance of L . brevis to hop compounds. Mol Microbiol, 2002 Oct, 46(2), 381 - 94 Domains in the S-layer protein CbsA of Lactobacillus crispatus involved in adherence to collagens, laminin and lipoteichoic acids and in self-assembly; Antikainen J et al.; The protein regions in the S-layer protein CbsA of Lactobacillus crispatus JCM 5810, needed for binding to collagens and laminin, anchoring to bacterial cell wall, as well as self-assembly, were mapped by deletion analysis of His-tagged peptides isolated from Escherichia coli and by heterologous expression on Lactobacillus casei . Mature CbsA is 410 amino acids long, and stepwise genetic truncation at both termini revealed that the region 32-271 carries the infor-mation for self-assembly of CbsA into a periodic structure . The lactobacillar S-layer proteins exhibit sequence variation in their assembly domain, but the border regions 30-34 and 269-274 in CbsA are conserved in valine-rich short sequences . Short deletions or substitutions at these regions affected the morphology of His-CbsA polymers, which varied from sheet-like to cylindrical tubular polymers, and further truncation beyond the DNA encoding residues 32 and 271 leads to a non-periodic aggregation . The self-assembly of the truncated peptides, as seen by electron microscopy, was correlated with their behaviour in a cross-linking study . The shorter peptides not forming a regular polymer were observed by the cross-linking study and mass spectrometry to form dimers, trimers and tetramers, whereas the other peptides were cross-linked to large multimers only . Binding of solubilized type I and IV collagens was observed with the His-CbsA peptides 1-274 and 31-287, but not with the smaller peptides regardless of their ability to form regular polymers . Strain JCM 5810 also adheres to immobilized laminin and, in order to analyse the possible laminin binding by CbsA, cbsA and its fragments were expressed on the surface of L . casei . Expression of the CbsA peptides 1-274, 1-287, 28-287 and 31-287 on L . casei conferred adhesiveness to both laminin and collagen immobilized on glass as well as to laminin- and collagen-containing regions in chicken colon and ileum . The C-terminal peptides 251-410 and 288-410 bound to L . crispatus JCM 5810 cells from which the S-layer had been depleted by chemical extraction, whereas no binding was seen with the His-CbsA peptides 1-250 or 1-269 or to cells with an intact S-layer . The His-CbsA peptides 251-410 and 288-410 bound to teichoic acids of several bacterial species . The results show that CbsA is an adhesive complex with an N-terminal assembly domain exhibiting affinity for pericellular tissue components and a cationic C-terminal domain binding to negatively charged cell wall components. Arzneimittelforschung, 2002, 52(9), 706 - 15 Local treatment of vaginal infections of varying etiology with dequalinium chloride or povidone iodine . A randomised, double-blind, active-controlled, multicentric clinical study; Petersen EE et al.; This randomised, double-blind, multicentric clinical study compared the efficacy and tolerability of the two vaginal antiseptics, 10 mg dequalinium chloride (CAS 522-51-0, Fluomycin N) and 200 mg povidone iodine (CAS 25655-41-8), in a parallel-group design . A total of 180 patients with vaginal infections of varying etiology participated in this study (bacterial vaginosis, fluor vaginalis, vulvo-vaginal candidiasis, trichomoniasis) . Patients were randomly allocated to one of the two treatment groups and were treated once per day for 6 days . Control examinations took place 5 to 7 days after the end of treatment, and 3 to 4 weeks after the first control examination . The total symptoms score, a summary score for the clinical symptoms, discharge, burning, pruritus, redness of vulva/vagina, was defined as primary efficacy parameter . The treatments at the first control examination were compared in the full analysis set using the Wilcoxon-Mann-Whitney U-test, 2-sided, thereby proving equivalence of both treatments at the 5% level . Both treatments strongly improved the symptoms of vaginal infections both on short-term and long-term follow-up . Descriptive analysis of the secondary parameters, vaginal pH, degree of purity of the vaginal flora, and number of lactobacilli in the wet mounts, supported the comparable efficacy of both therapies to restore the vaginal milieu . Analysis of the diagnostic subgroups indicated that irrespective of the diagnosis, both treatments improved the efficacy criteria as observed for the entire population . The global assessment of the therapeutic efficacy by investigators and patients supported the results of the efficacy analysis with good to very good ratings in 70-90% of the cases . A good tolerability of both preparations was observed in this study with a low number of adverse events in the test group (5.8%). Biosci Biotechnol Biochem, 2002 Sep, 66(9), 1963 - 6 Cytokine production by the murine macrophage cell line J774.1 after exposure to lactobacilli; Morita H et al.; Eleven strains of lactobacilli were tested for their ability to induce the murine macrophage-like cell line J774.1 to secrete cytokines . Some of the bacteria tested induce the production of interleukin(IL) 6, IL-12, and tumor necrosis factor a (TNF-alpha) by J774.1 cells . Seven strains also induced the production of IL-10 . However, no IL-1beta was produced . Lactobacillus acidophilus TMC 0356 significantly induced the production of more IL-6, IL-10, IL-12, and TNF-alpha than the other bacteria tested (p < 0.0001; ANOVA) . These results suggest that lactobacilli can activate macrophages to secrete both inflammatory and anti-inflammatory cytokines . Selected strains might be used to bring about pro or antiinflammatory immune reactions. Biosci Biotechnol Biochem, 2002 Sep, 66(9), 1835 - 43 Characterization of trehalose phosphorylase from Bacillus stearothermophilus SK-1 and nucleotide sequence of the corresponding gene; Inoue Y et al.; A bacterial trehalose phosphorylase (TPase; EC 2.4.1.64) was purified from the culture supernatant of Bacillus stearothermophilus SK-1 to apparent homogeneity, and some properties were investigated . Furthermore, a gene from SK-1 responsible for the TPase was cloned by Southern hybridization with a degenerate oligonucleotide probe synthesized on the basis of the N-terminal sequence of the purified enzyme . The Mr of the enzyme was estimated to be 150,000 by gel filtration and 83,000 by SDS-PAGE, so the enzyme is likely to be a homodimer . The enzyme had optimum activity at pH 7.0-8.0 or nearby and the optimum temperature was about 75 degrees C . The deduced amino acid sequence of the SK-1 TPase encodes a theoretical protein with a Mr of 87,950 . Alignment of amino acid sequences with a maltose phosphorylase from Lactobacillus brevis the crystal structure and active site of which had been analyzed suggested that these two phosphorylases evolved from a common ancestor . The Escherichia coli cells harboring the plasmid containing the cloned TPase gene had about 100 times the activity of SK-1. Caries Res, 2002 Sep-Oct, 36(5), 327 - 40 Evaluation of a computer program for caries risk assessment in schoolchildren; Hansel Petersson G et al.; The 'Cariogram' is an interactive PC program for caries risk evaluation . It takes into account the interactions between caries-related factors and expresses a graphic assessment of the risk . The aim of this study was to assess the caries risk in schoolchildren using the Cariogram and to evaluate the program by comparing the caries risk assessments with the actual change in DMF . A 2-year prospective study on 446 schoolchildren, 10-11 years old, was conducted . At baseline, data on general health, diet, oral hygiene and use of fluoride were obtained . Saliva analyses included mutans streptococci and lactobacilli counts, buffer capacity and secretion rate . DMFT and DMFS were calculated from records and bitewing radiographs . Scores were entered and caries risk was assessed . Re-examination for caries was done after 2 years . The children were divided into 5 groups according to the assessed caries risk at baseline . Where the Cariogram predicted a 0-20% (high risk), 21-40%, 41-60%, 61-80% and 81-100% (low risk) chance of avoiding new lesions, 8, 35, 42, 73 and 83%, respectively, had no new lesions 2 years later . Logistic regression analyses were carried out . When the Cariogram was included, only two factors, the Cariogram (p < 0.001) and the DMFS at baseline, i.e . past caries experience (p = 0.001), turned out to be significantly associated with caries increment . The Cariogram was the most powerful explanatory variable . When the Cariogram was excluded, lactobacillus count, mutans streptococci, diet intake frequency and DMFS at baseline were significantly associated with caries increment . The Cariogram predicted caries increment more accurately than any included single-factor model . How this finding can be translated into daily practice in the best and most practical way is a matter for future research . Int J Food Sci Nutr, 2002 Sep, 53(5), 367 - 73 Microbiological evaluation of ghanaian maize dough co-fermented with cowpea; Sanni AI et al.; Fermented maize dough meals form a large proportion of people's diet in Ghana . To enhance the nutritive value of these carbohydrate-rich foods, protein complementation was introduced . In this study, microbial ecology of fermenting maize dough fortified with 20% cowpea was investigated . A total of 106 microbial strains were isolated from different batches of the fermenting dough at periodic intervals . Ten genera of microorganisms namely Lactobacillus, Leuconostoc, Saccharomyces, Debaryomyces, Candida, Bacillus, Micrococcus, Klebsiella, Escherichia and Aspergillus were identified, with lactic acid bacteria species being predominant . A lactic count of log 9.9 cfu/g was obtained at the end of 72 h fermentation relative to log 6.6 cfu/g recorded for aerobic mesophiles . At the 12 h fermentation period, the population of yeasts was less than log 1.0 cfu/g, but gradually rose to log 5.36 cfu/g by 48 h followed a slight decline at the end of 72 h fermentation period (log 4.08 cfu/g) . Enteric microorganisms that were isolated from the raw cowpea were less than log 1.0 cfu/g at 12 h of fermentation, while the Aspergillus species were isolated from the raw maize and the dough subjected to drying treatment . The growth of inoculated enteropathogenic E . coli and S . typhimurium was inhibited in the cooked, fermented maize-cowpea dough, and at 72 h, they were not within detectable limit . The study concluded that addition of cowpea at 20% level did not affect the natural fermentation characteristics of the maize dough. J Urol, 2002 Nov, 168(5), 2236 - 9 Lactobacillus species is more cytotoxic to human bladder cancer cells than Mycobacterium Bovis (bacillus Calmette-Guerin); Seow SW et al.; PURPOSE: We determined if Lactobacillus species has growth inhibitory effects in human bladder cancer cell lines and how this effect compares with the known effects of Mycobacterium bovis, that is bacillus Calmette-Guerin (BCG) . MATERIALS AND METHODS: The growth of MGH and RT112 cells were determined by cell counts after 24, 48 and 72 hours of exposure to L . casei strain Shirota (Yakult, Singapore) or L . rhamnosus strain GG (National Collection of Industrial and Marine Bacteria, Ltd., Aberdeen, Scotland) (1 x 10 and 1 x 10 cfu) or BCG (1 x 10 cfu) in the presence and absence of streptomycin . Annexin-V was used to monitor the presence of pre-apoptotic cells . RESULTS: L . rhamnosus GG inhibited MGH proliferation and it was cytotoxic to RT112 cells (p <0.05) . L . casei Shirota was cytotoxic to the 2 cell lines (p <0.05) . BCG had a similar cytotoxic effect in MGH cells as Lactobacillus species but was not as effective in RT112 cells . Streptomycin abrogated the cytotoxic effect of Lactobacillus species but not that of BCG . Cytotoxic activity was not found in Lactobacilli culture supernates but it was induced in the presence of mammalian cells . L . rhamnosus GG induced apoptosis in RT112 but not in MGH cells . No apoptotic cells were detected after treatment with L . casei Shirota . CONCLUSIONS: Lactobacillus species induced cytotoxic effects in bladder cancer cells . Unlike BCG, it requires bacterial protein synthesis . Like BCG, L . casei Shirota induces cell death primarily via necrosis . The cytoxicity of these lactobacilli in bladder cancer cells raises the possibility of using this species of bacteria as intravesical agents for treating bladder cancer. J Pediatr Gastroenterol Nutr, 2002 Oct, 35(4), 527 - 31 Effect of Lactobacillus GG and breast-feeding in the prevention of rotavirus nosocomial infection; Mastretta E et al.; BACKGROUND: Rotavirus is one of the leading etiologic agents of nosocomial infections among children . The development of preventive measures is therefore important . The efficacy of GG in the treatment of rotavirus infection has been reported in literature, but there is only one recent study about its effectiveness in prevention of infection . The role of breast-feeding in the prevention of rotavirus infection is still debated . The aim of our study was to assess the efficacy of GG and breast-feeding in the prevention of nosocomial rotavirus infections . METHODS: In a randomized, placebo-controlled, double-blind study, 220 children aged 1 to 18 months hospitalized from December 1999 to May 2000, received GG (n = 114) at a dose of 10 colony-forming units or a comparable placebo (n = 106) every day of their hospital stay . Rotavirus testing on stool samples was performed for every patient on admission, during hospitalization, and after discharge . RESULTS: The total incidence of nosocomial rotavirus infections was 27.7% (61 of 220 patients) . The attack rate of rotavirus infections among the patients who received probiotic was 25.4% (29 of 114 patients), while for the placebo group it was 30.2% (32 of 106 patients) . The difference is not significant (P = 0.432) . Forty-seven of 220 infants (21.4%) were breast-fed, and 173 of 220 (78.6%) were non-breast-fed . The attack rate of rotavirus infections among breast-fed infants was 10.6% (5 of 47 infants), while for non-breast-fed infants it was 32.4% (56 of 173 infants) . The difference is significant (P = 0.003) . CONCLUSION: In our study, GG was ineffective in preventing nosocomial rotavirus infections, whereas breast-feeding was effective. J Biol Chem, 2002 Dec 27, 277(52), 50959 - 65 Epub 2002 Oct 21. Probiotic bacterium prevents cytokine-induced apoptosis in intestinal epithelial cells; Yan F et al.; Probiotic bacteria are microorganisms that benefit the host by preventing or ameliorating disease . However, little information is known regarding the scientific rationale for using probiotics as alternative medicine . The purpose of this paper is to investigate the mechanisms of probiotic beneficial effects on intestinal cell homeostasis . We now report that one such probiotic, Lactobacillus rhamnosus GG (LGG), prevents cytokine-induced apoptosis in two different intestinal epithelial cell models . Culture of LGG with either mouse or human colon cells activates the anti-apoptotic Akt/protein kinase B . This model probiotic also inhibits activation of the pro-apoptotic p38/mitogen-activated protein kinase by tumor necrosis factor, interleukin-1alpha, or gamma-interferon . Furthermore, products recovered from LGG culture broth supernatant show concentration-dependent activation of Akt and inhibition of cytokine-induced apoptosis . These observations suggest a novel mechanism of communication between probiotic microorganisms and epithelia that increases survival of intestinal cells normally found in an environment of pro-apoptotic cytokines. J Appl Microbiol, 2002, 93(5), 884 - 93 Characterization and selection of vaginal Lactobacillus strains for the preparation of vaginal tablets; Mastromarino P et al.; AIMS: To characterize and select Lactobacillus strains for properties that would make them a good alternative to the use of antibiotics to treat human vaginal infections . METHODS AND RESULTS: Ten Lactobacillus strains belonging to four different Lactobacillus species were analysed for properties relating to mucosal colonization or microbial antagonism (adhesion to human epithelial cells, hydrogen peroxide production, antimicrobial activity towards Gardnerella vaginalis and Candida albicans and coaggregation with pathogens) . The involvement of electrostatic interactions and the influence of bacterial metabolic state in the binding of lactobacilli to the cell surface were also studied . Adherence to epithelial cells varied greatly among the Lactobacillus species and among different strains belonging to the same Lactobacillus species . The reduction in surface negative electric charge promoted the binding of several Lactobacillus strains to the cell membrane whereas lyophilization reduced the adhesion capacity of many isolates . The antimicrobial activity of lactobacilli culture supernatant fluids was not directly related to the production of H2O2 . CONCLUSIONS: Three strains (Lactobacillus brevis CD2, Lact . salivarius FV2 and Lact . gasseri MB335) showed optimal properties and were, therefore, selected for the preparation of vaginal tablets . The selected strains adhered to epithelial cells displacing vaginal pathogens; they produced high levels of H2O2, coaggregated with pathogens and inhibited the growth of G . vaginalis . SIGNIFICANCE AND IMPACT OF THE STUDY: The dosage formulation developed in this study appears to be a good candidate for the probiotic prophylaxis and treatment of human vaginal infections. J Appl Microbiol, 2002, 93(5), 781 - 6 A collaborative study of a method for enumeration of probiotic enterococci in animal feed; Leuschner RG et al.; AIMS: Validation of an enumeration method to be used as an official control method in the framework of Council Directive 70/524/EEC for probiotic enterococci used as feed additives . METHODS AND RESULTS: Twenty laboratories in 12 European countries carried out a collaborative study . A plate count method using bile esculin azide (BEA) agar was used . Precision data in terms of repeatability (r) and reproducibility (R) of the method using different feeding stuffs and three inoculation levels were determined . Enterococci were present in the samples as a single component or in mixtures with other probiotic feed additives . The enumeration of enterococci on BEA agar showed a relative standard deviation (RSD)r of 1.5-3.6% and an RSD(R) between 2.9 and 7.4% . BEA agar was selective for enterococci in the presence of other probiotic micro-organisms such as pediococci, lactobacilli and yeast . CONCLUSIONS: For routine analysis of viable enterococci concentrations in feeding stuffs, the use of BEA is recommended . This methodology is not applicable for mineral feeds . SIGNIFICANCE AND IMPACT OF THE STUDY: An official control method for enumeration of authorized probiotic enterococci in feeding stuffs was validated . The results are intended for consideration for adoption as CEN and ISO standards. Lett Appl Microbiol, 2002, 35(5), 428 - 32 Glutamate uptake in Lactobacillus delbrueckii subsp . bulgaricus CNRZ 208 and its enhancement by a combination of Mn2+ and Mg2+; de Giori GS et al.; AIMS: To demonstrate the mechanism of glutamate uptake in the dairy strain Lactobacillus delbrueckii subsp . bulgaricus CNRZ 208, and to characterize key aspects of the system . METHODS AND RESULTS: Glutamate uptake proceeded via an active transport system requiring an exogenous source of energy . The system also transported aspartate and glutamine . It was unique, with a Kt of 2.8 micro mol l-1 and a Vmax of 900 micro mol s-1 (g dry weight)-1 . The activity was optimal at pH 7.3 and 50 degrees C, was independent of the glutamate charge, and was enhanced by Mn2+ + Mg2+ in combination . Inhibition of the activity by uncouplers and ionophores showed that transport was driven by an ATP-dependent mechanism involving the proton-motive force . This inhibition was partially abolished in the presence of both Mn2+ and Mg2+ . CONCLUSIONS: We demonstrated for the first time that an active transport system governs the uptake of the essential amino acid glutamate in Lact . delbrueckii subsp . bulgaricus CNRZ 208, the activity of which is enhanced by a combination of Mn2+ and Mg2+ . SIGNIFICANCE AND IMPACT OF THE STUDY: The potential of the findings is discussed with reference to the growth of Lact . delbrueckii subsp . bulgaricus in mixed-strain cultures for the dairy industry. Lett Appl Microbiol, 2002, 35(5), 370 - 4 Characterization of Lactobacillus plantarum from wine must by PCR species-specific and RAPD-PCR; Spano G et al.; AIMS: Physiological and molecular analysis such as PCR species-specific and randomly amplified polymorphic PCR (RAPD-PCR) have been used for typing of Lactobacillus plantarum strains from typical wine must . METHODS AND RESULTS: Phenotypic tests such as API 50CH and evaluation of D-L-lactate production from glucose were used to perform a preliminary characterization of lactobacilli . Furthermore, 18 strains of lactobacilli were analyzed by PCR species-specific oligonucleotides based on short sequences of the recA gene . CONCLUSIONS: Four strains were identified as belonging to the L . plantarum species and were further analysed by RAPD-PCR . The RAPD-PCR profiles were similar in all strains that had positive results for species-specific PCR, suggesting that the four L . plantarum strains were closely related . SIGNIFICANCE AND IMPACT OF THE STUDY: Using PCR species-specific as a preliminary screening test and then RAPD-PCR can be as considered the most reliable method of performing a rapid and correct typing of L . plantarum from wine must. J Gen Virol, 2002 Nov, 83(Pt 11), 2891 - 5 Interaction of the Cro repressor with the lysis/lysogeny switch of the Lactobacillus casei temperate bacteriophage A2; Ladero V et al.; The transcriptional switch region of Lactobacillus casei temperate bacteriophage A2 contains three similar 20 bp operator subsites, O(1), O(2) and O(3), which are interspersed between the divergent promoters P(R) and P(L) . The Cro protein binds initially to O(3), which overlaps the -35 region of P(L), excluding the RNA polymerase (sigma(A)-RNAP) from it . This results in the switching off of cI transcription and directs the incoming phage into the lytic cycle . At higher concentrations, Cro also binds to O(1) and/or O(2), which overlap P(R), probably introducing a bend in the intervening DNA . This interaction induces DNA looping, which provokes the subsequent displacement of sigma(A)-RNAP from P(R) . Consequently, Cro abolishes the binding of sigma(A)-RNAP to the genetic switch of A2 and, presumably, its own synthesis, contributing indirectly to the entry of phage development into its late stages. Obstet Gynecol . 2002 Oct;100(4):765. Douching in relation to bacterial vaginosis, lactobacilli, and facultative bacteria in the vagina; Ness RB et al.; OBJECTIVE: To study how frequency, recentness, and reason for douching impact bacterial vaginosis-related vaginal microflora and the occurrence of cervical pathogens . Douching has been linked to bacterial vaginosis as well as to chlamydial cervicitis in some, but not all, studies . METHODS: A total of 1200 women at high risk for sexually transmitted infections were enrolled from five clinical sites around the United States . Cross-sectional, structured interviews were conducted and vaginal swabs were self-obtained for Gram stain, culture, and DNA amplification tests for Neisseria gonorrhoeae and Chlamydia trachomatis . RESULTS: Douching at least once per month was associated with an increased frequency of bacterial vaginosis . Those who douched recently (within 7 days) were at highest risk {odds ratio (OR) 2.1, 95% confidence interval (CI) 1.3, 3.1} . Douching for symptoms (OR 1.7, 95% CI 1.1, 2.6) and for hygiene (OR 1.3, 95% CI 1.0, 1.9) both related to bacterial vaginosis risk . The associations between douching and Gardnerella vaginalis, Mycoplasma hominis, and lack of hydrogen peroxide-producing lactobacilli were similar to those between douching and bacterial vaginosis . Gonococcal or chlamydial cervicitis was not associated with douching . CONCLUSION: Douching for symptoms or hygiene, particularly frequent or recent douching, was associated with bacterial vaginosis and bacterial vaginosis-associated vaginal microflora, but not with gonococcal or chlamydial cervicitis. Appl Microbiol Biotechnol, 2002 Oct, 60(1-2), 73 - 80 Epub 2002 Aug 23. Production and stability of 3-hydroxypropionaldehyde in Lactobacillus reuteri; Luthi-Peng Q et al.; 3-Hydroxypropionaldehyde (3-HPA) is considered as a potent antimicrobial substance . Exploration of its application as a food preservative or as a therapeutic auxiliary agent has been documented in the literature . In the present work, factors that may impact on 3-HPA accumulation by Lactobacillus reuteri and on the stability of 3-HPA were investigated . Three media - H(2)O, milk and MRS broth - were chosen as test systems . Data indicated that 3-HPA accumulation in resting cells of L . reuteri in a two-step fermentation is greatly affected by temperature, pH, cell age and biomass as well as components in the test system . Within 2 h of incubation, 170 mM 3-HPA could be produced with a cell dry weight of 30 g/l, representing 85% of the glycerol supplied (200 mM) in H(2)O . The presence of glycerol during cell growth increased the productivity of 3-HPA by resting cells . In general, 3-HPA is much more stable in H(2)O than in milk and MRS . Factors that enhanced accumulation of 3-HPA did not simply show the same positive impact on the stability of 3-HPA . Thus, for defined applications, factors affecting production and stability of 3-HPA should be evaluated separately. Int J Food Microbiol, 2003 Jan 25, 80(2), 161 - 9 Fermentation by Lactobacillus fermentum Ogi E1 of different combinations of carbohydrates occurring naturally in cereals: consequences on growth energetics and alpha-amylase production; Calderon M et al.; Glucose, fructose, sucrose and starch are naturally present in cereals . Fermentation of different combinations of these carbohydrates by Lactobacillus fermentum Ogi E1, a sourdough heterofermentative lactobacillus, was investigated to determine effects on fermentation kinetics, growth energetics and alpha-amylase production . Irrespective of the substrate combination, the strain was able to simultaneously produce alpha-amylase and consume starch, glucose, fructose and sucrose . In mixtures of starch with either sucrose or fructose or with both fructose and glucose, yields of alpha-amylase from biomass (Y(amy/x)) were similar to those observed for starch . However, for starch and glucose or starch, glucose, fructose and sucrose mixtures, both Y(amy/x) and the specific rate of alpha-amylase production decreased markedly . In fructose- or sucrose-containing mixtures, mannitol was formed stoichiometrically indicating that fructose served as electron acceptor, and acetate was produced at constant yield from biomass (Y(ac/x)) (1 g acetate g biomass(-1)) . Acetate production was expected to confer to the strain a competitive advantage during natural fermentation by improving biomass formation and growth through an increase in the ATP gain . Y(ATP) varied depending on the carbohydrate mixture, indicating different effects of substrate mixtures on the efficiency in ATP coupling to biomass formation . Compared to starch fermentation, the highest value of Y(ATP) (29 g biomass mol ATP(-1)) was estimated for the starch/fructose mixture but no increase in mu(max) was observed . The lowest value (16 g biomass mol ATP(-1)) was obtained for the starch, glucose and fructose mixture, whereas for the mixture of all carbohydrates, Y(ATP) was similar to that obtained with starch alone (20 g biomass mol ATP(-1)) and it was intermediary for the starch and sucrose mixture (17 g biomass mol ATP(-1)) . It is concluded that competitiveness of the strain cannot be based on expected energy gain in mixed substrate fermentation involving fructose and sucrose with glucose and starch, but rather on its ability to simultaneously use carbohydrates while producing alpha-amylase and to produce acetic acid . Acetic acid production could enhance the strain capacity to inhibit nonacid-tolerant, competitive microflora at the earlier stage of natural fermentation. J Agric Food Chem, 2002 Oct 23, 50(22), 6497 - 500 Comparative survey in Lactobacillus plantarum of the growth and metabolism of arginine and citrulline in different media; Arena ME et al.; Arginine deiminase activity increased in the presence of arginine in Lactobacillus plantarum strains N4 and N8 isolated from orange . The influence of citrulline and ornithine on arginine deiminase and ornithine transcarbamylase activities was strain-dependent . The growth and arginine and citrulline metabolism of L . plantarum were studied in the presence of tomato juice . Its addition enhances the growth in both strains . The specific amino acids utilization was inversely proportional to the initial glucose concentration . Arginine and citrulline addition to basal medium exerted a stimulatory effect on the growth of N4 strain, and this effect was observed only with citrulline in strain N8 . The magnitude of this effect was lower in the presence of tomato juice. J Food Prot, 2002 Oct, 65(10), 1586 - 9 Behavior of Lactobacillus plantarum and Saccharomyces cerevisiae in fresh and thermally processed orange juice; Alwazeer D et al.; Lactobacillus plantarum and Saccharomyces cerevisiae are acid-tolerant microorganisms that are able to spoil citrus juices before and after pasteurization . The growth of these microorganisms in orange juice with and without pasteurization was investigated . Two samples of orange juice were inoculated with ca . 10(5) CFU/ml of each microorganism . Others were inoculated with ca . 10(7) CFU/ml of each microorganism and then thermally treated . L . plantarum populations were reduced by 2.5 and <1 log10 CFU/ml at 60 degrees C for 40 s and at 55 degrees C for 40 s, respectively . For the same treatments, S . cerevisiae populations were reduced by >6 and 2 log10 CFU/ml, respectively . Samples of heated and nonheated juice were incubated at 15 degrees C for 20 days . Injured populations of L . plantarum decreased by ca . 2 log10 CFU/ml during the first 70 h of storage, but those of S . cerevisiae did not decrease . The length of the lag phase after pasteurization increased 6.2-fold for L . plantarum and 1.9-fold for S . cerevisiae, and generation times increased by 41 and 86%, respectively . The results of this study demonstrate the differences in the capabilities of intact and injured cells of spoilage microorganisms to spoil citrus juice and the different thermal resistance levels of cells . While L . plantarum was more resistant to heat treatment than S . cerevisiae was, growth recovery after pasteurization was faster for the latter microorganism. Gut, 2002 Nov, 51(5), 659 - 64 Increased mucosal tumour necrosis factor alpha production in Crohn's disease can be downregulated ex vivo by probiotic bacteria; Borruel N et al.; BACKGROUND AND AIMS: Tumour necrosis factor alpha (TNF-alpha) plays a key role in the pathogenesis of intestinal inflammation in Crohn's disease . The effect of bacteria on TNF-alpha release by intestinal mucosa was investigated . METHODS: Ileal specimens were obtained at surgery from 10 patients with Crohn's disease (ileal stricture) and five disease controls undergoing right hemicolectomy (caecal cancer) . Mucosal explants from each specimen were cultured for 24 hours with either non-pathogenic Escherichia coli, Lactobacillus casei DN-114001, L bulgaricus LB10, or L crispatus (each study contained blank wells with no bacteria) . Tissue and bacterial viability was confirmed by lactate dehydrogenase (LDH) release and culture . Concentrations of TNF-alpha were measured in supernatants and the phenotype of the intestinal lymphocytes was analysed by flow cytometry . RESULTS: Coculture of mucosa with bacteria did not modify LDH release . Release of TNF-alpha by inflamed Crohn's disease mucosa was significantly reduced by coculture with L casei or L bulgaricus; changes induced by L crispatus or E coli were not significant . The effect of L casei and L bulgaricus was not prevented by protease inhibitors . Coculture with L casei and L bulgaricus reduced the number of CD4 cells as well as TNF-alpha expression among intraepithelial lymphocytes from Crohn's disease mucosa . None of the bacteria induced changes in non-inflamed mucosa . CONCLUSIONS: Probiotics interact with immunocompetent cells using the mucosal interface and modulate locally the production of proinflammatory cytokines. Electrophoresis, 2002 Sep, 23(19), 3321 - 31 Pulsed-field gel electrophoretic analysis of the genome of Lactobacillus gasseri ATCC33323, and construction of a physical map; Abs El-Osta YG et al.; Some species of Lactobacillus are of major industrial and health significance as fermenting agents in the manufacturing of food products, as food preservatives, as "probiotic" bacteria or as vaccine delivery vehicles . In spite of their importance, there is a paucity of published information on their genome organization and structure . In this study, a combination of pulsed field gel electrophoresis (PFGE) and hybridization approaches was used to investigate the genome of L . gasseri neotype strain ATCC33323 . PFGE analysis of chromosomal DNA (after digestion with the rare-cutting restriction enzymes I-CeuI, CspI, SmaI, ApaI, and SgrAI) allowed the chromosome size of L . gasseri to be estimated at 1.96 Mbp, and also revealed the presence of a linear plasmid of 48.5 kbp . A physical map of the L . gasseri chromosome, containing 6 sites for the enzymes I-CeuI and 12 for CspI, was constructed . Placed on the map were the genes dnaA and gyrB (usually located close to the origin of replication on the bacterial chromosome) and 18 ribosomal RNA (rrn) genes . Mapping analysis also revealed that the chromosome contained six rrn operons, and that one of them was inverted in orientation with respect to the others . Each rrn operon contained a single copy of each of the three rrn genes, 23S rRNA (rrl), 16S rRNA (rrs) and 55 rRNA (rrf) gene . The constructed physical map should be a useful foundation for genomic and genetic studies of the lactobacilli and provides a platform for applied research, such as the engineering of Lactobacillus strains with improved characteristics for industrial and probiotic applications. Sex Transm Dis, 2002 Oct, 29(10), 581 - 7 Rectal applications of nonoxynol-9 cause tissue disruption in a monkey model; Patton DL et al.; BACKGROUND: Efforts to develop topical microbicide products have all but ignored evaluation for rectal use . GOAL: The goal of this study was to assess the effects of multiple rectal applications of Conceptrol (containing 4% nonoxynol-9) on flora and mucosal tissues in the pig-tailed macaque model . STUDY DESIGN: Monkeys (8 per group) received daily rectal applications of Conceptrol, placebo gel, or no product, for 3 days . At each visit, a preapplication rectal lavage specimen and swab specimen for microbiology and pH determination were collected . Conceptrol or placebo gel (2.5 ml) was then administered intrarectally . Fifteen minutes after application, samples were again collected . RESULTS: Gross observation of rectal lavage indicated sheets of epithelium 15 minutes after application of the nonoxynol-9 product . Histopathology of these samples revealed epithelial sheets with stroma attached . The presence of H(2)O(2)-producing lactobacilli remained relatively constant, whereas that of H(2)O(2)-producing viridans streptococci diminished in all nonoxynol-9-exposed animals in which these organisms were detected at baseline . CONCLUSIONS: Repeated applications of nonoxynol-9 disrupts the rectal mucosa of the pig-tailed macaque . The disruption of these tissues could have serious implications for an increase in likelihood of acquisition of sexually transmitted infection/HIV in humans. J Dairy Res, 2002 Aug, 69(3), 457 - 72 Screening and selection of Lactobacillus strains for use as adjunct cultures in production of semi-hard cheese; Antonsson M et al.; Thirty-three Lactobacillus strains were tested as adjuncts in a cheese model system . Eighteen strains originated from cheese (nine Lactobacillus spp . and nine Lb . paracasei/casei) and 15 from human intestinal mucosa (11 Lb . rhamnosus; three Lb . paracasei; one Lb . plantarum) . Model cheeses weighing 120 g were made of cheese grains from full-scale production of washed curd semi-hard cheese (Herrgard) . The model system was reproducible and similar to full-scale production with respect to moisture, salt content, pH and microbial flora . The model cheeses were sampled for aerobic and anaerobic plate count and viable counts of Lactobacillus and Lactococcus . The presence of adjuncts in the model cheeses was confirmed by typing isolates with Randomly Amplified Polymorphic DNA (RAPD) . The sensory properties of model cheeses were described . In a first trial 23 of the 33 adjuncts were re-isolated from the corresponding model cheeses after 9 or 13 weeks . Adjuncts of Lb . paracasei were re-isolated more frequently than adjuncts of Lb . rhamnosus . Nine strains were selected, on the basis of their ability to grow and be a dominating part of the microflora of model cheese with interesting sensory properties . These strains were further studied together with two commercial cultures . The sensory influences on model cheeses of six of the adjuncts were confirmed, and flavour scores were in the range of 2.9-7.1 for model cheeses with different adjuncts while the control had a flavour score of 5.6 (0-10 scale) . Survival and growth of seven out of the nine strains correlated with the results of the first trial . Growth and influence on flavour of four adjunct cultures were confirmed in experimental cheese manufactured in a 400-1 open vat. Antonie Van Leeuwenhoek, 2002 Aug, 82(1-4), 333 - 40 Anti-hypertensive activity of fermented dairy products containing biogenic peptides; Takano T; A considerable amount of research has been conducted to study the health benefits of fermented milks . One possible mechanism for the beneficial effects is the involvement of biogenic compounds, which are produced by the action of microorganisms and make the products directly beneficial without the need for live bacteria . Research on the health effects of a pasteurized sour milk, which is fermented by a starter culture containing Lactobacillus helveticus as the predominant microorganism, indicated clear evidence for the involvement of biogenic compounds . In this paper, beneficial effects of sour milk are reviewed with special attention to the effect on anti-hypertension and angiotensin-I converting enzyme inhibitory peptides identified in sour milk. Antonie Van Leeuwenhoek, 2002 Aug, 82(1-4), 263 - 9 Respiration capacity and consequences in Lactococcus lactis; Gaudu P et al.; We recently reported that the well-studied fermenting bacterium Lactococcus lactis could grow via a respirative metabolism in the presence of oxygen when a heme source is present . Respiration induces profound changes in L . lactis metabolism, and improvement of oxygen tolerance and long-term survival . Compared to usual fermentation conditions, biomass is approximately doubled by the end of growth, acid production is reduced, and large amounts of normally minor end products accumulate . Lactococci grown via respiration survive markedly better after long-term storage than fermenting cells . We suggest that growth and survival of lactococci are optimal under respiration-permissive conditions, and not under fermentation conditions as previously supposed . Our results reveal the uniqueness of the L . lactis respiration model . The well-studied 'aerobic' bacteria express multiple terminal cytochrome oxidases, which assure respiration all throughout growth; they also synthesize their own heme . In contrast, the L . lactis cydAB genes encode a single cytochrome oxidase (bd), and heme must be provided . Furthermore, cydAB genes mediate respiration only late in growth . Thus, lactococci exit the lag phase via fermentation even if heme is present, and start respiration in late exponential phase . Our results suggest that the spectacularly improved survival is in part due to reduced intracellular oxidation during respiration . We predict that lactococcal relatives like the Enterococci, and some Lactobacilli, which have reported respiration potential, will display improved survival under respiration-permissive conditions. BMC Microbiol . 2001;1(1):20. Utilization of tmRNA sequences for bacterial identification; Schonhuber W et al.; BACKGROUND: Ribosomal RNA molecules are widely used for phylogenetic and in situ identification of bacteria . Nevertheless, their use to distinguish microorganisms within a species is often restricted by the high degree of sequence conservation and limited probe accessibility to the target in fluorescence in situ hybridization (FISH) . To overcome these limitations, we examined the use of tmRNA for in situ identification . In E . coli, this stable 363 nucleotides long RNA is encoded by the ssrA gene, which is involved in the degradation of truncated proteins . RESULTS: Conserved sequences at the 5'- and 3'-ends of tmRNA genes were used to design universal primers that could amplify the internal part of ssrA from Gram-positive bacteria having low G+C content, i.e . genera Bacillus, Enterococcus, Lactococcus, Lactobacillus, Leuconostoc, Listeria, Streptococcus and Staphylococcus . Sequence analysis of tmRNAs showed that this molecule can be used for phylogenetic assignment of bacteria . Compared to 16S rRNA, the tmRNA nucleotide sequences of some bacteria, for example Listeria, display considerable divergence between species . Using E . coli as an example, we have shown that bacteria can be specifically visualized by FISH with tmRNA targeted probes . CONCLUSIONS: Features of tmRNA, including its presence in phylogenetically distant bacteria, conserved regions at gene extremities and a potential to serve as target for FISH, make this molecule a possible candidate for identification of bacteria. Biotechnol Prog, 2002 Sep-Oct, 18(5), 1033 - 8 The application of a generic feedstock from wheat for microbial fermentations; Wang R et al.; The feasibility of a generic fermentation feedstock produced from wheat flour has been confirmed in several fermentations of yeasts, bacterium, and filamentous fungus for the production of commodity chemicals . Saccharomyces cerevisiae was incubated for the observation of yeast growth and ethanol production, Pichia farinosa for glycerol production, Monascus purpureus for fungal growth and pigment production, and Lactobacillus bulgaricus for bacteria growth and lactic acid production . The results confirmed that the feedstock contained no inhibitory components to the strains tested . Similar or higher metabolite yields were obtained in comparison with other studies carried in commonly used media. Int J Syst Evol Microbiol, 2002 Sep, 52(Pt 5), 1881 - 5 Carnobacterium viridans sp . nov., an alkaliphilic, facultative anaerobe isolated from refrigerated, vacuum-packed bologna sausage; Holley RA et al.; A facultatively anaerobic, non-spore-forming, psychrophilic, gram-positive, non-aciduric but alkaliphilic, rod-shaped bacterium (MPL-11T) was found to be responsible for green discoloration of refrigerated vacuum-packaged bologna upon opening of the package . Although Aerococcus viridans, which had been implicated earlier in causing the same problem, was also found, this is the first report of discoloration caused by an organism shown to be a species of Carnobacterium . Bacterial discoloration was caused by H2O2 production upon exposure of the meat to air . Strain MPL-11T is catalase- and oxidase-negative . It is not motile and does not reduce nitrate to nitrite or produce ammonia from arginine . It does not grow in acetate-containing broth or agar (Rogosa) or produce H2S . The peptidoglycan is of the meso-diaminopimelic acid type and it produces predominantly L(+)-lactic acid from glucose . It grows from at least 2 to 30 degrees C over a pH range from 5.5 to 9.1 . Ribotyping suggested that strain MPL-11T could be a species of either Lactobacillus or Carnobacterium, but analysis using DNA sequences from the 16S rRNA gene showed conclusively that the organism belonged to the genus Carnobacterium . Since acid is not produced from amygdalin, inulin, mannitol, methyl alpha-D-glucoside or D-xylose, the organism differs from the seven described species of Carnobacterium . In addition, strain MPL-11T is the first member of the genus found that does not produce acid from ribose . It is capable of acid production/growth on galactose, glucose, fructose, mannose, N-acetylglucosamine, aesculin, cellobiose, maltose, lactose, sucrose, trehalose and tagatose . Although extremely salt tolerant, it does not grow in > or = 4% NaCl . On the basis of phenotypic and genotypic data, it is concluded that this isolate represents a separate, novel species . Accordingly, the name Carnobacterium viridans sp . nov . is proposed . The type strain is strain MPL-11T (= ATCC BAA-336T = DSM 14451T). Int J Syst Evol Microbiol, 2002 Sep, 52(Pt 5), 1745 - 8 Lactobacillus pantheris sp . nov., isolated from faeces of a jaguar; Liu B et al.; Two novel Lactobacillus strains were isolated from the faeces of a jaguar in Beijing Zoo . They were gram-positive, non-spore-forming, non-motile, rod-shaped organisms that exhibited no catalase activity . The strains fermented cellobiose, D-fructose, galactose, D-glucose, lactose, maltose, D-mannose, salicin, trehalose, N-acetyl glucosamine, beta-gentiobiose and D-tagatose . D(-)-Lactic acid was the exclusive product from glucose fermentation . The G+C content of the DNA of strain A24-2-1T was 52.7 mol % . Phylogenetic analysis based on 16S rDNA sequence similarity indicated that the strains represented a novel member of the genus Lactobacillus and were most closely related to Lactobacillus manihotivorans LMG 18010T, with 16S rDNA sequence similarity of 95.2% . The DNA-DNA relatedness between them was 13% . The name Lactobacillus pantheris sp . nov . is proposed for the novel strains, with strain A24-2-1T as the type strain (= AS 1.2826T = LMG 21017T). Proc Natl Acad Sci U S A, 2002 Oct 15, 99(21), 13437 - 41 Epub 2002 Oct 01. X-ray structure of a bifunctional protein kinase in complex with its protein substrate HPr; Fieulaine S et al.; HPr kinase/phosphorylase (HprK/P) controls the phosphorylation state of the phosphocarrier protein HPr and regulates the utilization of carbon sources by Gram-positive bacteria . It catalyzes both the ATP-dependent phosphorylation of Ser-46 of HPr and its dephosphorylation by phosphorolysis . The latter reaction uses inorganic phosphate as substrate and produces pyrophosphate . We present here two crystal structures of a complex of the catalytic domain of Lactobacillus casei HprK/P with Bacillus subtilis HPr, both at 2.8-A resolution . One of the structures was obtained in the presence of excess pyrophosphate, reversing the phosphorolysis reaction and contains serine-phosphorylated HPr . The complex has six HPr molecules bound to the hexameric kinase . Two adjacent enzyme subunits are in contact with each HPr molecule, one through its active site and the other through its C-terminal helix . In the complex with serine-phosphorylated HPr, a phosphate ion is in a position to perform a nucleophilic attack on the phosphoserine . Although the mechanism of the phosphorylation reaction resembles that of eukaryotic protein kinases, the dephosphorylation by inorganic phosphate is unique to the HprK/P family of kinases . This study provides the structure of a protein kinase in complex with its protein substrate, giving insights into the chemistry of the phospho-transfer reactions in both directions. Syst Appl Microbiol, 2002 Aug, 25(2), 228 - 31 Differentiation of Lactobacillus helveticus, Lactobacillus delbrueckii subsp bulgaricus, subsp lactis and subsp delbrueckii using physiological and genetic tools and reclassification of some strains from the ATCC collection; Delley M et al.; Several physiological tests of glucose metabolism and genetic tools including species specific probes and 16S rDNA sequences were used to identify strains of L . helveticus and the group of L . delbrueckii with its three subspecies lactis, bulgaricus, and delbrueckii . These species are important for the milk industry as fermenting lactic acid bacteria . The identification procedure was applied to the different strains of these species available from the ATCC collection and allowed to reclassify part of them. J Pediatr Gastroenterol Nutr, 2002 Sep, 35(3), 350 - 5 Lactobacillus plantarum reduces the in vitro secretory response of intestinal epithelial cells to enteropathogenic Escherichia coli infection; Michail S et al.; OBJECTIVE: Enteropathogenic (EPEC) is a Gram-negative bacillus that causes diarrhea . Secretory responses of intestinal epithelial cells can be seen after EPEC infection . Probiotics, which are live bacteria that have proven benefit to the host, play a role in the treatment and prevention of the different enteric pathogens . The goals of the study were to determine whether the probiotic agent (LBP) strain 299v alters the secretory changes seen in EPEC infection and, if so, what underlying mechanism is possible . METHODS: Caco-2 cell monolayers were rapidly infected with EPEC strain E2348/69 and immediately mounted in Ussing chambers . The monolayers were exposed to LBP before, after, and simultaneously with EPEC infection . Short circuit current (Isc) was measured in the Ussing chamber . RESULTS: EPEC infection caused an increase in short circuit current that was reduced by preincubation with LBP ( < 0.01) . No direct bactericidal effect was observed, but LBP reduced the attachment of EPEC to Caco-2 cells . CONCLUSION: LBP can play an important role in reducing the secretory change in response to EPEC infection, possibly through inhibition of its binding . However, the presence of the probiotic agent before the infection is necessary . In this setting, its role is more preventive rather than therapeutic. FEMS Microbiol Lett, 2002 Sep 10, 214(2), 257 - 61 Effects of oxidoreduction potential combined with acetic acid, NaCl and temperature on the growth, acidification, and membrane properties of Lactobacillus plantarum; Ouvry A et al.; The effects of oxidoreduction potential (Eh) combined with acetic acid, NaCl and temperature on the growth, acidification, and membrane properties of Lactobacillus plantarum were studied . The culture medium was set at pH 5, and two different Eh values were adjusted using nitrogen (Eh = +350 mV) or hydrogen (Eh = -300 mV) gas . In reducing condition, the growth was slowed and the acidification delayed at 37 degrees C, but not at 10 degrees C . A synergistic inhibitory effect of reducing Eh, acetic acid and NaCl was observed, mainly for delaying the lag phase before acidification . These results may be explained by changes in ATPase activity, membrane fluidity and surface properties. Arch AIDS Res, 1996, 10(4), 239 - 41 Alternative treatment for bacterial vaginosis in pregnant patients; restoration of vaginal acidity and flora; Tasdemir M et al.; PIP: In Turkey, physicians treated 22 first-trimester pregnant women aged 20-34 exhibiting signs and symptoms of bacterial vaginosis with commercially available yogurt containing more than 100 million Lactobacillus acidophilus per ml and at a pH of less than 4.5 . The treatment involved vaginal douching with a 10-ml syringe daily for 7 days and was repeated after a 1-week interval . The signs and symptoms used as criteria for bacterial vaginosis included subjective symptoms of foul-smelling discharge, vaginal pH more than 4.7, presence of clue cells in the vaginal discharge, and foul odor caused by the addition of 10% potassium hydroxide to the vaginal discharge (i.e., positive amine test) . Cure was defined as the disappearance of 3 of the 4 criteria 4-6 weeks after completing the 2nd treatment and clinical improvement (i.e., patients' subjective feelings on 3rd day of treatment) . All the women were considered clinically improved on the 3rd day of treatment . 4 weeks after the 2nd treatment, 20 (90%) women exhibited none of the criteria for bacterial vaginosis . At 6 weeks, 19 (86.4%) had no criteria . These findings suggest that commercially available yogurt may restore the microenvironment and pH of the vagina, thus representing a viable treatment of bacterial vaginosis . This treatment may be preferable during pregnancy, since the use of any drug during pregnancy should be minimized . It will reduce the likelihood of a premature delivery . Health Stat Nord Ctries, 1985 Jan-Feb, 36(1), 77 - 84 {Vaginal microscopic evidence in women with symptoms of genital infection}; Cerbini S et al.; PIP: Vaginal living organisms have been defined as the most complete in human ecology . As many as 100 million colonies of microorganisms can live in a single ml of vaginal liquid . More common are the anaerobic types, particularly the lactobacillus and bacteroides . The low pH of the vagina, normally 4, is due to the high lactic acid content of the lactobacillus . Pathogenic bacteria is inhibited at 4.5 pH . Conversely, it proliferates at a high pH . The slightest alteration of this delicate equilibrium allows potentially pathogenic bacteria to grow to a concentration high enough to produce symptoms . Researchers have sought to identify new pathological microorganisms, due to the increase in reported cases of vaginal infection in recent years . New strains such as microplasmas and chlamydia are assuming greater importance . Evidence of the numerous vaginal microbes comes from anamnestic, symptomatological data and from simple laboratory tests conducted in gynecological clinics with microscopes and Gram coloration . 663 women were examined in 6 laboratories . The computerized data revealed that only 29, or 4.37%, showed no symptoms at all and had gone to the lab for a mere check up; 175, or 26.40%, complained of pain, burning and itching; 361, or 54.45%, revealed symptoms associated with leukorrhea; 98, or 14.78%, showed only leukorrhea . The epidemiological study shows the importance of candida albicans in the infectious pathology of the female genital organ . Trichomonas vaginalis and grambacteria infection have been detected particularly when an IUD is used . The negative result of numerous bacterioscopic exams has shown the necessity of more in-depth study of genital infections, using analytical cultural methodology . Appl Environ Microbiol, 2002 Oct, 68(10), 5155 - 9 In vitro adhesion specificity of indigenous Lactobacilli within the avian intestinal tract; Edelman S et al.; In vitro adherence of Lactobacillus strains to cell and tissue types along the chicken alimentary tract and to ileal mucus were determined . Fresh isolates from chickens adhered to the epithelium of crop and, in a strain-dependent manner, to follicle-associated epithelium and the apical surfaces of mature enterocytes of intestinal villi . No adherence to the apical surfaces of undifferentiated enterocytes, the mucus-producing goblet cells, or the ileal mucus was detected. Contracept Technol Update, 1999 Mar, 20(3), 25 - 7 Vaginal microbicides needed for female-controlled prevention; Paediatrics Forum . Acute diarrhoea; PIP: Acute diarrhea is still responsible for about 40% of diarrhea-associated deaths, and oral rehydration therapy (ORT) does not actually reduce the duration of diarrhea . A species of lactobacilli specific for the human gut was first isolated in 1987, Lactobacillus casei strain GG, and several trials have used colonization of the gut by this organism as an adjunct to ORT . A placebo-controlled, triple-blind study in Pakistan showed a significant reduction in the number of children with persistent watery stools at 48 hours, as well as a reduction in stool output and vomiting . Dioctahedral smectite (DS) has been found to adsorb viruses, bacteria, and toxins resulting in the protection of gut mucosa . A randomized double-blind trial (placebo-controlled) studied outcome in 90 males, 3-24 months old, with acute diarrhea of or= 5 days duration . After rehydration, patients were given either 1.5 g of DS or placebo 4 times a day for 3 days . At 48 hours, 42% of the treatment group were free from diarrhea, as opposed to 13% of the placebo group, and at 3.5 days 20% of the placebo group still had diarrhea, as opposed to none in the treatment group . Mean duration of diarrhea in the treatment groups was 54.1 hours (placebo 72.9 hours, p 0.001) . However, mean stool output was similar (97.9 g/kg vs . 110.9 g/kg) . Bismuth subsalicylate (BSS) has been frequently used in adults with benefits in both prevention and treatment . 142 Chilean children 4-36 months old were randomized to receive either placebo or BSS (100 mg/kg/day) 5 times a day for 5 days . Stools were normal in the E . coli group by 72 hours as opposed 139 hours in the placebo group (p 0.01), while rotavirus-infected stools normalized in 57.5 hours, as opposed to 104.5 hours in the placebo group . Other effective approaches include micronutrient supplementation including zinc and folate . Shengzhi Yu Biyun, 1990 Feb, 10(1), 38 - 42 {The effect of contraceptive suppository of morrhuatic acid on lactobacilli in the vagina}; Wu JY et al.; PIP: The positive incidence of vaginal lactobacilli after culture was 69.7% in 33 volunteers tested by the quantitative method . Those positive patients were divided into different groups for study . 21 used morrhuatic acid and become group 1 . Their log10 X+or- Se of lactobacilli was 7.9545 +or- 0.2748 before the suppository was used, 8.3422 +or- 0.1910/12 hours after suppository use, and the paired t value was 2.4327 . The geometric mean of lactobacilli was significantly higher at 12 hours after treatment (p0.05) . The 2nd group of 17 subjects who were sexually active had levels of quantitative lactobacilli of 7.7435 +or- 0.2968/12 hours after sexual activity, and no statistical significance were found although that was lower than 8.0587 +or- 0.2129 prior to sexual activity . The 6 subjects who were sexually active and who were tested both before and after the use of morrhuatic acid served as the 3rd group . There the levels of lactobacilli before and after treatment (with sexual activity) were quite close, 8.2475 +or- 0.0891 and 8.0219 +or- 0.2840, respectively . Results revealed that the vaginal lactobacilli were not affected by the morrhuatic acid suppository . (author's modified) Cent Eur J Public Health, 2002 Sep, 10(3), 97 - 9 The Lactobacilli--Candida relationship in cervico-vaginal smears; Demirezen S; The aim of this study is to determine whether there is a relationship between the presence of Lactobacilli and the growth of Candida . For this purpose, 1110 cervico-vaginal smears were examined cytologically and 450 patients who were normally menstruating were subsequently selected among them . Fifty nine of 450 were diagnosed as having Candida and they were accepted as study group . Three hundred and ninety-one patients who were diagnosed as having no infectious agents were accepted as control group . Lactobacilli were present in 50 of 59 in various degree and were absent in 9 of 59 . The prevalence rates of the presence of Lactobacilli (84.74%) as well as intermediate cells (98.30%) were higher in study group than in control group . There was also a significant correlation between the presence of Lactobacilli and the growth of Candida (p < 0.05) . Statistical analysis was performed with chi 2 test using SPSS package programme . We can suggest that Lactobacilli do not have a protective role when there is Candida growth in the vaginal mucosa. J Biochem Mol Biol, 2002 Mar 31, 35(2), 244 - 7 Mutations within the putative active site of heterodimeric deoxyguanosine kinase block the allosteric activation of the deoxyadenosine kinase subunit; Park I et al.; Replacement of the Asp-84 residue of the deoxyguanosine kinase subunit of the tandem deoxyadenosine kinase/ deoxyguanosine kinase (dAK/dGK) from Lactobacillus acidophilus R-26 by Ala, Asn, or Glu produced increased Km values for deoxyguanosine on dGK . However, it did not seem to affect the binding of Mg-ATP . The Asp-84 dGK replacements had no apparent effect on the binding of deoxyadenosine by dAK . However, the mutant dGKs were no longer inhibited by dGTP, normally a potent distal endproduct inhibitor of dGK . Moreover, the allosteric activation of dAK activity by dGTP or dGuo was lost in the modified heterodimeric dAK/dGK enzyme . Therefore, it seems very likely that Asp-84 participates in dGuo binding at the active site of the dGK subunit of dAK/dGK from Lactobacillus acidophilus R-26. CCL Family Found . 1991 May-Jun;17(6):3. What about NFP? Unknown risks; Oral contraceptives et al.; PIP: The effects of oral contraceptives on serum folate and hematologic levels in 526 women were studied by a new radioassay method and by Lactobacillus casei assay . Serum folate levels were not significantly lower compared with a control group (p more than .1 by L casei assay; p more than .2 by radioassay) . Hemoglobin values and iron deficiency anemia were significantly less in the control group (p less than .001) . It was concluded that oral contraceptives do not cause folate deficiency anemia . Female patient, 1979 May, 4(5), 29 - 32, 34 Human milk for the premature infant; Cunningham MD et al.; PIP: Closer study of premature infants has led to the establishment of human milk as the recommended nutritional standard for them . Infants fed with human milk are found to gain protection against necrotizing enterocolitis, and have fewer upper respiratory tract infections and systemic infections . Antibacterial components of human milk include leukocytes (macrophages, granulocytes, and T and B cell lymphocytes), the B cell secreted immunoglobulins, and complement factors . Lactoperoxidase and lactoferrin in breast milk are particularly active against group B beta hemolytic streptococcus and staphylococcus, respectively . Fresh human milk helps in the colonization of the infant's gut by Lactobacillus bifidus, a bacteria that suppresses other possibly pathogenic organisms . Nutritional values of human milk offer certain advantages and disadvantages over cow's milk . The quality of human milk protein is superior to that of cow's milk, and human milk's higher levels of cystine are especially required in the premature infant, where the liver is deficient in the enzyme required to convert methionine to cystine . Human milk is low in certain essential electrolytes and trace elements, such as sodium . It can also harbor drugs, pollutants, and harmful viruses that can be dangerous to the infant . Freezing of human milk for banking can destroy many of the immunological advantages of its use . Given that the advantages seem to outweigh the disadvantages, the authors favor the use of human milk for premature infants . Biomed Environ Sci, 2002 Jun, 15(2), 153 - 65 Effects of anti-caries antibodies on Lactobacillus GG in its fermentation and storage periods; Wei H et al.; OBJECTIVE: To investigate how antibodies influence the fermentation of Lactobacillus GG and how Lactobacillus GG influences the biological properties of antibodies during the fermentation and storage periods . METHODS: Anti-caries immune colostrum powder (IP) and control colostrum powder (CP), skimmed milk powder (SP) at concentrations of 1%, 2.5%, 5% and 10% (w/v) were added to MRS and 50 mM Hepes buffer system was used in the milk, the growth curves of Lactobacillus GG including viable cells, lactic acid concentratrion and pH, and also the titer of specific antibodies were determined during the fermentation and storage periods . RESULTS: In MRS, SP could improve the growth of Lactobacillus GG in all periods of fermentation, especially at the concentrations of 5% and 10% . CP at the concentrations of 1% and 2.5% had a good initial velocity at the beginning and CP (1%, 2.5%, 5%, 10%) in all the groups could reach a high viable cell concentration at the end of fermentation, which suggested that there were some growth factors for Lactobacillus GG in CP overcoming the inhibition of unspecific antibodies . IP at 1%, 2.5%, 5% and 10% could inhibit the growth of Lactobacillus GG in all the fermentation periods, the critical concentration point was between 2.5%-5%, and there was a competition between growth factors and the inhibition of specific antibodies . In pasteurized milk, the influence of Hepes could help the fermentation start quickly and reach the log phase earlier than in the control group; however, when combined with 5% IP, the growth of Lactobacillus GG was strongly inhibited during all the fermentation periods . The fermentation of Lactobacillus GG had no significant effect on the titer change of anti-caries antibodies during the fermentation and storage periods . CONCLUSION: SP and CP is beneficial to the growth of Lactobacillus GG in MRS, whereas there is a competition between growth factors and the inhibition of specific antibodies to the growth of Lactobacillus GG, the critical concentration of IP is 2.5%-5% . Hepes could influence the titer of antibodies through buffering Ph, thus influence the fermentation of Lactobacillus GG . The fermentation of Lactobacillus GG with IP or CP could be used for the development of functionally fermented immune milk in the future. Spec Care Dentist, 2002 May-Jun, 22(3), 103 - 7 Risk factors for dental caries in children with cerebral palsy; dos Santos MT et al.; This study was conducted to examine the oral condition and the salivary and microbiological parameters associated with dental caries in 62 children with cerebral palsy, who came from households of low socioeconomic status (Study Group) . This group had mixed (6 to 11 years old) and permanent (11 to 16 years old) dentition . Dental examinations were performed to measure dental caries, plaque index, salivary levels of mutans streptococci and lactobacilli, salivary flow rate, pH of stimulated saliva, and buffer capacity of saliva . A group of 67 non-handicapped children from similar socioeconomic backgrounds also were examined using these parameters (Control Group) . Data were analyzed statistically by non-parametric tests and by correlation . The results showed that children with cerebral palsy who had permanent dentitions had a higher mean decayed, missing and filled surfaces index, as well as a higher plaque index for both sexes . Microbiological examination revealed higher levels of mutans streptococci among Study Group subjects with mixed dentition than in the Control Group . Also, lactobacillus counts were higher in the Study Group, regardless of sex or dentition . With respect to salivary flow rate, pH and buffering capacity, lower mean values were obtained for the Study Group. Inflammation, 2002 Oct, 26(5), 207 - 14 Signalling mechanisms involved in the induction of inducible nitric oxide synthase by Lactobacillus rhamnosus GG, endotoxin, and lipoteichoic acid; Korhonen R et al.; BACKGROUND AND AIMS: Probiotic Lactobacillus rhamnosus GG (Lactobacillus GG) has been found beneficial in the treatment of viral and antibiotic-associated diarrhea . Recently, it has also been shown to induce nitric oxide (NO) production, and have some other immunostimulatory effects . The aim of the present study was to investigate the mechanisms involved in the induction of inducible nitric oxide synthase (iNOS) and NO production by Lactobacillus GG . METHODS AND RESULTS: iNOS expression and NO production induced by Lactobacillus GG, lipopolysaccharide (LPS), and lipoteichoic acid (LTA) was abrogated by NOS inhibitors L-NMMA and 1400W, by a protein synthesis inhibitor cycloheximide, by a tyrosine kinase inhibitor genistein and by a NF-kappaB inhibitor pyrrolidinedithiocarbamate (PDTC) in J774 macrophages . Polymyxin B inhibited NO production induced by LPS, but did not inhibit Lactobacillus GG induced NO production . p42/44 MAP-kinase inhibitor PD98059, dexamethasone and cyclosporine A inhibited partially iNOS protein expression and NO formation in Lactobacillus GG, LPS and LTA treated cells . Ro 31-8220 (protein kinase C inhibitor) and SB203580 (p38 MAP-kinase inhibitor) had only a minor effect on NO production . CONCLUSIONS: Lactobacillus GG induced NO production through iNOS pathway and the mechanisms mediating that process were very similar with those involved in LPS and LTA induced NO synthesis. Am J Obstet Gynecol, 2002 Sep, 187(3), 561 - 8 Antimicrobial components of vaginal fluid; Valore EV et al.; OBJECTIVE: We examined the antimicrobial activity and composition of vaginal fluid . STUDY DESIGN: Vaginal fluid from preweighed tampons was assayed for pH, lactic acid, and antimicrobial polypeptides . The fluid was also fractionated by molecular filtration . Antimicrobial activity of whole fluid was determined against representative resident and exogenous microbes, and its fractions were tested against Escherichia coli . RESULTS: Vaginal fluids (5/5 donors) were permissive for Lactobacillus crispatus and vaginalis and Candida albicans, but not for Escherichia coli, Streptococcus group B, and Lactobacillus jensenii in three of five donors . The antimicrobial activity against E coli was predominantly in a <3-kd fraction and correlated with both low pH and high lactic acid content . Compared with a matched pH buffer, lactic acid markedly suppressed the growth of E coli . Concentrated 2- or 5-fold, the protein-rich fraction was active against E coli . CONCLUSION: Vaginal fluid exerts selective antimicrobial activity against nonresident bacterial species . The activity is mediated by lactic acid, low pH, and antimicrobial polypeptides. Chem Pharm Bull (Tokyo), 2002 Sep, 50(9), 1233 - 7 Development of a simple HPLC method for determination of paeoniflorin-metabolizing activity of intestinal bacteria in rat feces; He JX et al.; A simple and reproducible HPLC method for the determination of paeoniflorin (PF)-metabolizing activity of intestinal bacteria in rat feces was developed and validated . Orally administered PF, a major active constituent of Paeoniae Radix, is metabolized into a bioactive compound, paeonimetabolin I (PM-I) by intestinal bacteria . Direct determination of the PF-metabolizing rate into PM-I is hard to achieve by HPLC due to the lack of intense chromophore in PM-I . However, when PF was incubated with Lactobacillus brevis, an intestinal bacterium, in the presence of phenylmercaptan, the metabolizing rate of PF into 8-phenylthio-paeonimetabolin I (PT-PM-I) was found to be equivalent to that of PF into PM-I . Thus, the PF-metabolizing activity of intestinal bacteria in rat feces was determined by measuring the rate of biotransformation of PF into PT-PM-I, which was detected by HPLC at 255 nm . This method can be utilized in the biopharmaceutical study of traditional Chinese formulations containing Paeoniae Radix. J Agric Food Chem, 2002 Sep 25, 50(20), 5605 - 11 Fermentability of grape must after inhibition with dimethyl dicarbonate (DMDC); Delfini C et al.; Dimethyl dicarbonate (DMDC) was added to grape must and to synthetic media and results showed that, at 20 degrees C, 150 mg.L(-)(1) DMDC completely inhibited the fermentation of a grape must that was previously inoculated with 10(6) cells.mL(-)(1) Saccharomyces bayanus and Saccharomyces uvarum . Brettanomyces intermedius, Candida guilliermondii, Hansenula jadinii, Hansenula petersonii, Kloeckera apiculata, Pichia membranaefaciens, and Saccharomyces cerevisiae were inhibited by 250 mg.L(-)(1) . Candida valida was inhibited in the presence of 350 mg.L(-)(1), whereas Hanseniaspora osmophila, Saccharomycodes ludwigii, Schizosaccharomyces pombe, and Zygosaccharomyces bailii required 400 mg.L(-)(1) . Delay of fermentation (but not inhibition) was noted in the presence of 400 mg.L(-)(1) for the following cultures: Brettanomyces anomalus, Hanseniaspora uvarum, Metschnikowia pulcherrima, Schizosaccharomyces japonicus, Torulaspora delbrueckii, and Zygosaccharomyces florentinus . Acetobacter aceti and Lactobacillus sp . were completely inhibited using 1000 and 500 mg.L(-)(1) DMDC, respectively . The fermentation of a grape must inoculated with 10(6) cells.mL(-)(1) of different wine yeasts was delayed for 4 days after the prior addition of 200 mg.L(-)(1) of DMDC; 200 mg.L(-)(1) DMDC did not show any residual inhibitory effect after 12 h, nor did 300 mg.L(-)(1) 24 h after the addition . In cellar experiments, indigenously contaminated grape musts (with and without skins) showed a delay in fermentation of 48 h after the addition of only 50 mg.L(-)(1) DMDC . The possibility of using DMDC (as pure grade as commercially available) in grape must as a disinfectant for the decontamination of musts indigenously contaminated with wild yeast should be considered seriously, despite its apparent low solubility in water. Anal Biochem, 2002 Sep 1, 308(1), 1 - 4 Application of vitamin B(12)-targeting site on Lactobacillus helveticus B-1 to vitamin B(12) assay by chemiluminescence method; Sato K et al.; Lactobacillus helveticus B-1 is assumed to have a vitamin B(12)-targeting (or B(12)-binding) site on the cells, since the binding reaction of vitamin B(12) with L . helveticus B-1 cells proceeded instantly and quantitatively . This reaction is specific to complete B(12) compounds, cobalamins, and can be used for a vitamin B(12) assay method by chemiluminescence . The calibration graph was linear from 0.1 to 10.0 ng/mL . The B(12) contents in oyster and sardine were 75.9 and 39.4 microg/100g, respectively . These values were very close to those obtained using a chemilumi-ADVIA Centaur immunoassay system with intrinsic factor and to those obtained by microbiological assays. J Appl Microbiol, 2002, 93(4), 714 - 24 Influence of pH, temperature and culture media on the growth and bacteriocin production by vaginal Lactobacillus salivarius CRL 1328; Juarez Tomas MS et al.; AIMS: To study the influence of pH, temperature and culture medium on the growth and bacteriocin production by vaginal Lactobacillus salivarius subsp . salivarius CRL 1328 . METHODS AND RESULTS: The study was performed using a complete factorial experimental design . Lactobacillus salivarius was cultivated in LAPTg and MRS broths, adjusted to specific initial pH, and at different temperatures of incubation . The growth, which was evaluated by the Gompertz model, was higher in MRS broth than in LAPTg broth . The initial pH of the culture medium and the temperature had a dramatic effect on the production of bacteriocin . The optimal conditions for bacteriocin production were different to those for optimal growth . The decrease in the pH of the culture medium was parallel to the growth; pH had similar final values in both the MRS and the LAPTg broths . CONCLUSIONS: The optimal growth conditions were recorded in MRS broth, with an initial pH of 6.5 and a temperature of 37 degrees C . The maximum bacteriocin activity was obtained in LAPTg after 6 h at 37 degrees C, and at an initial pH of 6.5 or 8.0 . SIGNIFICANCE AND IMPACT OF THE STUDY: The application of a complete factorial design, and the evaluation of the growth parameters through the Gompertz model, enabled a rapid and simultaneous exploration of the influence of pH, temperature and growth medium on both growth and bacteriocin production by vaginal Lact . salivarius CRL 1328. J Appl Microbiol, 2002, 93(4), 566 - 76 Antifungal activities of two Lactobacillus plantarum strains against Fusarium moulds in vitro and in malting of barley; Laitila A et al.; AIMS: The Lactobacillus plantarum strains VTT E-78076 (E76) and VTT E-79098 (E98) were studied for their antifungal potential against Fusarium species . METHODS AND RESULTS: In vitro screening with automated turbidometry as well as direct and indirect impedimetric methods clearly showed Lact . plantarum cell-free extracts to be effective against Fusarium species including Fusarium avenaceum, F . culmorum, F . graminearum and F.oxysporum . However, great variation in growth inhibition was observed between different Fusarium species and even between strains . The antifungal potential of Lact . plantarum E76 culture, including cells and spent medium, was also examined in laboratory-scale malting with naturally contaminated two-rowed barley from the crops of 1990-96 . The growth of the indigenous Fusarium flora was restricted by the addition of Lact . plantarum E76 to the steeping water . However, the antifungal effect was greatly dependent on the contamination level and the fungal species/strains present on barley in different years . CONCLUSIONS: Lactobacillus plantarum strains E76 and E98 had a fungistatic effect against different plant pathogenic, toxigenic and gushing-active Fusarium fungi . SIGNIFICANCE AND IMPACT OF THE STUDY: The present study indicates that Lact . plantarum strains with known and selected characteristics could be used as a natural, food-grade biocontrol agent for management of problems caused by Fusarium fungi during germination of cereals. Int J Food Microbiol, 2002 Oct 25, 78(3), 217 - 26 Antimicrobial effect of natural preservatives in a cooked and acidified chicken meat model; Lemay MJ et al.; The inhibitory effect of Microgard 100, Microgard 300, nisin, Alta 2002, Perlac 1902, sodium lactate and essential oil of mustard on microorganisms experimentally inoculated was screened in an acidified chicken meat model (pH = 5.0) and stored for 2 weeks at a none restrictive growth temperature of 22 degrees C . All antimicrobials tested were used at the highest concentration recommended by their manufacturer . Sausage batter made with mechanically deboned chicken was inoculated with a mixed culture of Escherichia coli ATCC 25922, Brochothrix thermosphacta CRDAV452, and a protective culture Lactobacillus alimentarius BJ33 (FloraCan L-2) . A final cell concentration of 3-4 log CFU g (-1) was targeted after cooking at a core temperature of 55 degrees C for each microorganism in order to assess cell count variation effectively . Composition, water activity (a(w)), pH and redox potential of the sausage model was also evaluated . The E . coli population decreased steadily during storage and was close or below detection level (< 1 log CFU g (-1)) for all treatments, including the control, after 14 days . Sodium lactate was most effective against B . thermosphacta; population was 4 log lower than the control after 14 days of storage . When essential oil of mustard was used, aerobic mesophilic bacteria and lactic acid bacteria were significantly lower than the control after 2 days of storage (P < or = 0.05) . The other antimicrobial agents tested had no significant effect on the aerobic mesophilic bacteria, E . coli, B . thermosphacta and lactic acid bacteria counts, when compared to the control. Appl Microbiol Biotechnol, 2002 Sep, 59(6), 665 - 71 Epub 2002 Jul 13. The mannitol dehydrogenase gene (mdh) from Leuconostoc mesenteroides is distinct from other known bacterial mdh genes; Aarnikunnas J et al.; The N-terminal amino acid sequences of the intact protein and three tryptic peptides from a 41 kDa protein purified from a commercial mannitol dehydrogenase (MDH) enzyme preparation of Leuconostoc mesenteroides ATCC-9135 were determined . Oligonucleotides deduced from these peptide sequences were used to isolate the putative mdh gene from L . mesenteroides using the Vectorette system . Nucleotide sequence analysis revealed an open reading frame (ORF1) of 1,014 bp encoding a putative MDH protein of 338 amino acids, and another open reading frame (ORF2) encoding an unknown protein of 245 amino acids . In Northern blots, a transcript of approximately 2.2-kb was detected with an mdh-specific probe . Mapping of the 5'-end of the 2.2-kb transcript indicated that mdh was the first gene of the operon . After fusion of six histidine codons to the 3'-end of the mdh gene and expression in Escherichia coli M15, active MDH was isolated using HisTrap purification . The overexpressed enzyme showed high specificity for mannitol and fructose . In dot blot hybridisation, the L . mesenteroides mdh-specific probe bound strongly to chromosomal DNA of Leuconostoc pseudomesenteroides and weakly to DNA of some heterofermentative Lactobacillus strains, whereas no hybridisation signals were obtained with DNA derived from strains carrying characterised mdh genes . Furthermore, the amino acid sequence similarity between L . mesenteroides MDH and other known MDHs was very low, suggesting that MDHs from heterofermentative lactic acid bacteria form a structurally and functionally separate enzyme group . Interestingly, L . mesenteroides MDH shared significant sequence similarity with the medium-chain dehydrogenase/reductase protein family. Nahrung, 2002 Aug, 46(4), 251 - 5 Utilization of Lactobacillus sp . for steroid glycoalkaloids degradation by lactic acid fermentation; Vesela M et al.; The degradation fo steroid glycoalkaloids (SGAs) has been studied in model solutions . The number of colony forming units (CFU) was determined using a nondirect (cultivation) method during all stages of fermentation . The changes in SGAs content were observed by HPLC on the Supelcosil LC-NH2 column . The changes in alpha-tomatine concentration added to fermented Lactobacillus MRS broth have been studied . A mathematical model of steroid glycoalkaloids degradation during lactic fermentation was proposed . The mathematical model was based on the experimental data of SGA and glucose concentration and should be used for study and prediction of SGA concentration changes of fermented samples . The ratio of SGA degradation rate by fermentation and by lactic acid hydrolysis was calculated . The experimental data evaluated by proposed mathematical model for the selected strain of Lactobacillus plantarum 976H show real feasibility for SGA degradation by lactic acid fermentation. J Dairy Res, 2002 May, 69(2), 255 - 67 Lactobacillus reuteri DSM 20016: purification and characterization of a cystathionine gamma-lyase and use as adjunct starter in cheesemaking; De Angelis M et al.; A homo-tetrameric approximately 160-kDa cystathionine gamma-lyase was purified to homogeneity from Lactobacillus reuteri DSM 20016 by four chromatographic steps . The activity was pyridoxal-5'-phosphate dependent and the enzyme catalyzed the alpha,gamma-elimination reaction of L-cystathionine, producing L-cysteine, ammonia and alpha-ketobutyrate . The enzyme was active towards a range of amino acids and amino acid derivatives, including methionine . The pH and temperature optima were found to be 8.0 and 35 degrees C . respectively . Isoelectric pH (pI) was approximately 5.0 as determined by two-dimensional electrophoresis . Sensitivity to chemical inhibitors was typical of lactococcal cystathionine gamma- and beta-lyases, except it was inhibited by sulphydryl reagents . The N-terminal sequence was MKFNTQLIHGGNSED, which had 100% homology with cystathionine beta-lyase of Lb . reuteri 104R (Accession Number (CAC05298) . Lb . reuteri DSM 20016 . together with 10 other strains of non-starter lactic acid bacteria, was used as adjunct starter in the production of miniature ('anestrato Pugliese-like cheeses . After 40 d ripening, the water-soluble extract of the cheeses with added Lactobacillus fermentum DT41 and Lb . reuteri DSM 20016 contained the highest enzyme activities on cystathionine and methionine substrates . Determinations of methanethiol, dimethyl sulphide, dimethyl disulphide and dimethyl trisulphide in the miniature cheeses confirmed the findings of enzyme activities. J Dairy Res, 2002 May, 69(2), 213 - 25 Microbial composition, including the incidence of pathogens, of goat milk from the bergamo region of italy during a lactation year; Foschino R et al.; Sixty samples of raw goat milk intended for Caprino cheese-making were collected from ten farms in the Bergamo area over a 6-month period . Analyses of main microbial groups, somatic cell count (SCC) and pH were performed to determine the effect of origin (farm) and lactation period (April - September) on microbial composition and the incidence of pathogens in milk . Overall mean values were: standard plate count (SPC), 5.0 x 10(4) cfu/ml; yeasts, 2.5 x 10(2) cfu/ml; coliforms, 91 x 10(2) cfu/ml; Escherichia coli, 2.9 cells/ml: enterococci, 1.1 x 10(2) cfu/ ml; lactococci, 3 4 x 10(3) cfu/ml; lactobacilli, 3.0 x 10(3) cfu/ml; halotolerant bacteria, 8.2 x 10(3) cfu/ml; spores of mesophilic aerobic bacteria, 11 cfu/ml; SSC, 9.9 x 10(5) cells/ml; pH, 6.63 . Moulds and spores of sulphite-reducing clostridia were found intermittently . Neither Salmonella spp . nor Listeria monocytogenes was detected, while Esch . coli O157: H7 was isolated from one milk sample (an incidence of 1.7%) . Staphylococcus aureus was discovered at a level > 10(2) cfu/ml in 26 samples (43%) with an overall mean of 12 x 10(3) cfu/ml, whereas coagulase-negative staphylococci were found in 54 samples (90%) with an overall mean of 1.3 x 10(3) cfu/ml . Of Staph . aureus strains, 23% proved to be enterotoxinogenic with a prevalence of enterotoxin C producers . Staph . caprae was the coagulase-negative species most frequently isolated; none of the coagulase-negative staphylococci strains synthesized any of the enterotoxins tested for . Sample source was the major factor affecting the microbial composition of goat milk: significant differences (P < 0.01) were observed among samples from different farms for SPC, coliforms, lactococci, lactobacilli and halotolerant bacteria . Period of lactation had a significant effect (P < 0.025) on SCC and pH . SPC correlated well with coliforms, lactococci and lactobacilli; SSC did not reveal positive interactions with any microbial groups or pH. Pharmacotherapy, 2002 Sep, 22(9), 1180 - 2 Lactobacillus species as a cause of ventilator-associated pneumonia in a critically ill trauma patient; Wood GC et al.; Lactobacillus species are ubiquitous inhabitants of the human gastrointestinal and genitourinary tracts and rarely cause infections . Pneumonia caused by Lactobacillus has been reported only in immunocompromised patients and in one patient with structural lung disease . A 39-year-old, immunocompetent, critically ill woman developed ventilator-associated pneumonia, which was found to be caused by Lactobacillus . To our knowledge, this is the first such report . The infection was treated successfully with 14 days of intravenous vancomycin . Based on this case, Lactobacillus should be considered a possible cause of ventilator-associated pneumonia in immunocompetent, critically ill patients. Odontostomatol Trop, 2002 Jun, 25(98), 19 - 26 Caries prevalence and some caries related factors for 12 year-old children from Vientiane and Luang Prabang provinces in Lao People's Democratic Republic; Tayanin GL et al.; Caries prevalence and some caries related factors for 100 twelve-year-old children from Vientiane and Luang Prabang provinces in Lao People's Democratic Republic were studied . The caries prevalence was high showing a DMFT of 4.6 with 9% of the children being caries free, the D comportment contributed mostly to the DMFT and the Significant Caries Index (Sic Index) was DMFT 8.0 . As to the caries related factors, 25% of the children were in the high mutans class, 31% of the group had high or very high Lactobacilli scores and 84-88% studied took local sweet snacks, chips and sweet drinks every day . Taking into consideration the high caries prevalence, the mutans streptococci distribution, the frequent intake of sweets and also the relatively low fluoride concentration in the drinking waters, oral health preventive programmes are indicated to prevent a possible caries epidemic in Laos in the future. Oral Dis, 2002 Jul, 8(4), 207 - 17 Bacterial and yeast flora of root surface caries in elderly, ethnic Chinese; Shen S et al.; OBJECTIVES: Root caries is emerging as a significant problem in the middle aged and elderly . As little data is available on the microbiology of root caries in Chinese cohorts, we evaluated 30 such lesions in elderly, institutionalized, ethnic Chinese . METHODS: Samples of carious dentine were aseptically taken from root caries lesions of 18 subjects (five males and 13 females, mean age 79.67 +/- 8.57) . The cultivable bacteria, both aerobic and anaerobic, were isolated and identified using standard methods and commercial identification kits . The yeasts were isolated on Sabouraud's agar and identified using the API system . RESULTS: The main findings were: (1) of the total isolates, 91.09% were Gram-positive and 8.91% were Gram-negative microorganisms; (2) the proportions of cocci and rods were 36.68 and 63.31%, respectively; (3) the predominant groups of organisms isolated were Streptococcus spp., Lactobacillus spp., Staphylococcus spp . and Actinomyces spp . with isolation frequencies of 100, 90.00, 73.33 and 63.33%, respectively; (4) the isolation frequency of yeasts belonging to Candida spp . (63.33%) was notably high although the proportion of yeasts within each sample was low (0.01%) . Candida dubliniensis, a newly identified yeast species particularly prevalent in HIV infection, comprised 14.29% of yeasts . CONCLUSIONS: This study provides baseline information on the microbiologic features of root caries in the elderly, ethnic Chinese . Whilst our data on the most predominant bacteria isolated in root caries concur with those from other regions of the world the isolation of C . dubliniensis from these lesions has not been documented before. J Bacteriol, 2002 Oct, 184(19), 5518 - 23 Cold shock induction of the cspL gene in Lactobacillus plantarum involves transcriptional regulation; Derzelle S et al.; Fragments of the cspL promoter region were fused to the gusA reporter and reintroduced into Lactobacillus plantarum cells, either on multicopy plasmids or through single-copy chromosomal integration . beta-Glucuronidase activity and primer extension data demonstrate that the cspL promoter is induced in response to cold shock and that multicopy constructs quench the induction of the resident cspL gene. J Infect, 2002 Aug, 45(2), 122 - 7 Bacteria in two-millennia-old cheese, and related epizoonoses in Roman populations; Capasso L; A tremendous volcanic eruption destroyed all the life around Mount Vesuvius during the night between 24 and 25 August, 79 AD . Two famous towns, Pompeii and Herculaneum, were completely buried under volcanic products . At Herculaneum, about 25m of volcanic mud killed about 250 people who had fled to the beaches in an attempt to escape (Bisel, S . C.,Rivista di Studi Pompeiani, 1, 123-124, 1987) . An anthropological examination of the skeletons of these "fugitives" reveals the bone lesions typical of brucellosis in 17.4% of adults (Capasso, L., International Journal of Osteoarchaelogy, 9, 277-288, 1999) . This very high incidence of brucellosis was theoretically linked to the consumption of ovine milk and its derivates, which is also indicated by both literary and figurative sources . A single carbonized cheese was found in Herculaneum; its analysis clearly reveals the excellent state of preservation of the milk curds . For the first time, we demonstrate the presence of a variety of bacteria, possibly Lactobacillus, that also includes cocco-like forms that seem to be morphologically and dimensionally consistent with Brucella . The long interval spent by the organic remains under the volcanic mud and high temperatures they suffered preclude the possibility of identifying the bacteria through molecular methods. Br J Nutr, 2002 Sep, 88 Suppl 1, S101 - 8 Competition for adhesion between probiotics and human gastrointestinal pathogens in the presence of carbohydrate; Lee YK et al.; The adhesion of Lactobacillus rhamnosus GG to human enterocyte-like Caco-2 cells was not inhibited by eight carbohydrates tested, namely N-acetyl-glucosamine, galactose, glucose, fructose, fucose, mannose, methyl-alpha-D-mannopyranoside and sucrose . The degree of hydrophobicity predicted the adhesion of L . rhamnosus GG to Caco-2 cells . L . rhamnosus GG, however, was able to compete with Escherichia coli and Salmonella spp . of low hydrophobicity and high adhesin-receptor interaction for adhesion to Caco-2 cells . The interference of adhesion of these gastrointestinal (GI) bacteria by L . rhamnosus GG was probably through steric hindrance, and the degree of inhibition was related to the distribution of the adhesin receptors and hydrophobins on the Caco-2 surface . A Carbohydrate Index for Adhesion (CIA) was used to depict the binding property of adhesins on bacteria surfaces . CIA was defined as the sum of the fraction of adhesion in the presence of carbohydrates, with reference to the adhesion measured in the absence of any carbohydrate . The degree of competition for receptor sites between Lactobacillus casei Shirota and GI bacteria is a function of their CIA distance . There were at least two types of adhesins on the surface of L . casei Shirota . The study provides a scientific basis for the screening and selection of probiotics that compete with selective groups of pathogens for adhesion to intestinal surfaces . It also provides a model for the characterisation of adhesins and adhesin-receptor interactions. Arch Latinoam Nutr, 2002 Mar, 52(1), 29 - 34 {The nutritional status change the effectiveness of a dietary supplement of lactic bacteria on the emerging of respiratory tract diseases in children}; Rio ME et al.; One hundred children 6 to 24 month old, normal or undernourished according to weight for height index, received during three months--autumn to winter--a dietary supplement of live Lactobacillus acidophilus and Lactobacillus Casei, 10(7)-10(8)/ml in fermented milk (LB) or an equivalent amount of fluid milk (L) as control . Children's follow-up was performed as outpatients in the Hospital Posadas (Great Buenos Aires) . Episodes of respiratory tract infections were recorded and classified according to severity as: Pneumonia (N); Bronchitis (B), Recurrent Obstructive Bronchitis (BOR) and upper respiratory tract infections (CVAS) . 58% of children fitted the study protocol, 22 in the LB and 36 in the L group; 21 were undernourished and 37 presented normal weight/height . No deaths were recorded . Total episodes were 103: 34 in LB and 69 in L, that means a frequency of 1.55 and 1.92 episodes/children respectively . In LB a maximum of 3 episodes/children was recorded, meanwhile the number reached 7 in L (p = 0.0373) . Severity was higher in L than LB: 0.06 vs . 0 for N; 0.69 vs . 0.45 for B + BOR and 1.17 vs . 1.09 for CVAS . In the control group frequency of severe pathologies was about twice in undernourished than in normal: 0.08 vs . 0.04 for N; 1.08 vs . 0.50 for B + BOR; no difference was found for CVAS . Live lactobacillus supplement suppressed pneumonia and decreased bronchitis in undernourished as well as in normal . In this study undernutrition not only increased the chance of suffering severe acute respiratory tract infections but also impaired the effectiveness of the supplement to decrease severity . The effect is explained on the basis of the immunocompetence depression linked to an inadequate nutritional status. Vaccine, 2002 Sep 10, 20(27-28), 3304 - 9 Protection against tetanus toxin after intragastric administration of two recombinant lactic acid bacteria: impact of strain viability and in vivo persistence; Grangette C et al.; Non-pathogenic lactic acid bacteria (LAB) are attractive as live carriers to deliver protective antigens to the mucosal immune system . Both persisting and non-persisting strains of lactic acid bacteria have been evaluated and seem to work equally well by the systemic and nasal routes of administration . However, it is not known if persistence and viability of the strain play a critical role when immunizing by the oral route . To address this question, recombinant LAB strains, able to persist (Lactobacillus plantarum NCIMB8826/pMEC127) or not (Lactococcus lactis MG1363/pMEC46) in the gastro-intestinal tract of mice and producing equivalent amounts of the tetanus toxin fragment C (TTFC) were compared to each other . A very strong ELISA TTFC-specific and protective humoral response was elicited by either live or UV-inactivated recombinant Lb . plantarum strains . In