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Aims Klebsiella pneumoniae , an important opportunistic pathogen of nosocomial inflection, is known for its ability to form biofilm. The purpose of the current study is to assess how co- or mono-cultured probiotics affect K. pneumoniae 's ability to produce biofilms and investigate the potential mechanisms by using a polyester nonwoven chemostat and a Caco-2 cell line. Methods and Results Compared with pure cultures of Lactobacillus rhamnosus and Lactobacillus sake , the formation of K. pneumoniae biofilm was remarkably inhibited by the mixture of L. rhamnosus, L. sake , and Bacillus subtilis at a ratio of 5:5:1 by means of qPCR and FISH assays. In addition, Lactobacillus in combination with B. subtilis could considerably reduce the adherence of K. pneumoniae to Caco-2 cells by using inhibition, competition, and displacement assays. According to the RT-PCR assay, the adsorption of K. pneumoniae to Caco-2 cells was effectively inhibited by the co-cultured probiotics, leading to significant reduction in the expression of proinflammatory cytokines induced by K. pneumoniae. Furthermore, the HPLC and RT-PCR analyses showed that the co-cultured probiotics were able to successfully prevent the expression of the biofilm-related genes of K. pneumoniae by secreting plenty of organic acids as well as the second signal molecule (c-di-GMP), resulting in inhibition on biofilm formation. Conclusion Co-culture of L. sake, L. rhamnosus , and B. subtilis at a ratio of 5:5:1 could exert an antagonistic effect on the colonization of pathogenic K. pneumoniae by down-regulating the expression of biofilm-related genes. At the same time, the co-cultured probiotics could effectively inhibit the adhesion of K. pneumoniae to Caco-2 cells and block the expression of proinflammatory cytokines induced by K. pneumoniae. [ABSTRACT FROM AUTHOR]

Aim The present study is a single-centre, randomized, controlled clinical trial aimed to evaluate the effectiveness of the probiotic Lacticaseibacillus rhamnosus TOM 22.8 (DSM 33500) strain, orally administrated, to treat vaginal dysbiosis. Methods and Results Overall, 80 women, with signs and symptoms of vaginal dysbiosis, were enrolled and allocated to the treatment group (A, n =60), who took 1 capsule of the probiotic strain for 10 consecutive days, or the non-treatment group (B, n =20), who did not receive any treatment. Clinical (vaginal signs and symptoms; pH of the vaginal fluid; Amsel criteria; Nugent score; Lactobacillary grade) and microbiological examinations were performed at baseline (T0), 10 days (T1), and 30 (T2) days after the oral administration of the probiotic TOM 22.8 strain. The latter resulted in a restoration of the physiological pH, accompanied by remission or attenuation of clinical signs and symptoms as well as the improvement of the quality of life (QoL). Microbiological data revealed a significant reduction of potentially pathogenic bacteria. Conclusion The administration of the L. rhamnosus TOM 22.8 probiotic strain could be proposed as an effective strategy for the treatment of vaginal dysbiosis. [ABSTRACT FROM AUTHOR]

Aims To investigate the effect of Lactobacillus rhamnosus on viral replication and cellular response to human rhinovirus (HRV) infection, including the secretion of antiviral and inflammatory mediators from well-differentiated nasal epithelial cells (WD-NECs). Methods and results The WD-NECs from healthy adult donors (N  = 6) were cultured in vitro , exposed to different strains of L. rhamnosus (D3189, D3160, or LB21), and infected with HRV (RV-A16) after 24 h. Survival and adherence capacity of L. rhamnosus in a NEC environment were confirmed using CFSE-labelled isolates, immunofluorescent staining, and confocal microscopy. Shed virus and viral replication were quantified using TCID50 assays and RT-qPCR, respectively. Cytotoxicity was measured by lactate dehydrogenase (LDH) activity. Pro-inflammatory mediators were measured by multiplex immunoassay, and interferon (IFN)-λ1/3 was measured using a standard ELISA kit. Lactobacillus rhamnosus was able to adhere to and colonize WD-NECs prior to the RV-A16 infection. Lactobacillus rhamnosus did not affect shed RV-A16, viral replication, RV-A16-induced IFN-λ1/3 production, or LDH release. Pre-exposure to L. rhamnosus , particularly D3189, reduced the secretion of RV-A16-induced pro-inflammatory mediators by WD-NECs. Conclusions These findings demonstrate that L. rhamnosus differentially modulates RV-A16-induced innate inflammatory immune responses in primary NECs from healthy adults. [ABSTRACT FROM AUTHOR]

Aims Clostridioides difficile  infections (CDI) are a major cause of morbidity and mortality in hospitalized patients. A probiotic formulation (Bio-K+) comprised of  Lactobacillus acidophilus  CL1285, Lacticaseibacillus casei  LBC80R, and Lacti. rhamnosus  CLR2 strains have been shown to reduce the incidence of CDI and antibiotic-associated diarrhea (AAD). This research aims to therefore elucidate the mechanism of action of the three probiotic strained against  C. difficile R20291, independently of the acidification of the environment.  Methods and Results Antitoxin activity was evaluated using ELISA method and the expression of  C. difficile  genes was evaluated using transcriptomic analysis in co-culture assays conducted in a bioreactor allowing precise control of the pH. The fermentation results demonstrated a decrease for toxin A and many genes directly related to  C. difficile  virulence were underexpressed in the co-cultures. Conclusions The lactobacilli tested could have a role in the motility, the quorum sensing (QS), the survival of the spores, and the germination potential of the spores, which are essential elements for the virulence of  C. difficile. [ABSTRACT FROM AUTHOR]

While conventional bacterial pneumonia mainly centralizes avoidance of bacterial colonization, it remains unclear how to restore the host immunity for hyperactive immunocompetent primary and immunocompromised secondary bacterial pneumonia. Here, probiotic-based nanoparticles of OASCLR were formed by coating chitosan, hyaluronic acid and ononin on living Lactobacillus rhamnosus. OASCLR nanoparticles could effectively kill various clinic common pathogens and antibacterial efficiency was >99.97%. Importantly, OASCLR could modulate lung microbiota, increasing the overall richness and diversity of microbiota by decreasing pathogens and increasing probiotic and commensal bacteria. Additionally, OASCLR could target inflammatory macrophages by the interaction of OASCLR with the macrophage binding site of CD44 and alleviate overactive immune responses for hyperactive immunocompetent pneumonia. Surprisingly, OASCLR could break the state of the macrophage's poor phagocytic ability by upregulating the expression of the extracellular matrix assembly, immune activation and fibroblast activation in immunocompromised pneumonia. The macrophage's phagocytic ability was increased from 2.61% to 12.3%. Our work provides a potential strategy for hyperactive immunocompetent primary and immunocompromised secondary bacterial pneumonia. [ABSTRACT FROM AUTHOR]

Introduction and aims We aim to investigate the lysate of Lactobacillus rhamnosus GG (LGG), a bacterium previously established as a gut probiotic, and to examine its postulated ability to enhance and restore human skin barrier function. Compromised skin barriers are implicated in various skin conditions including atopic dermatitis. Furthermore, skin barrier function decreases with age leading to dryness and sensitivity. Previous research has shown promising effects of LGG lysates on keratinocytes, including increasing transepidermal electrical resistance, increasing tight junction protein expression, cell migration promotion and mitigation of Staphylococcus aureus toxicity. This study aims to take a closer look at the molecular mechanisms behind the effect of LGG lysates on the skin, including terminal differentiation, lamin dispersal and nuclear degradation in keratinocytes. Methods Normal human epithelial keratinocytes and rat epithelial keratinocytes were grown and treated with the LGG lysate. Western blot analysis was used to determine the relative concentrations of tight junction proteins and other differentiation markers. A phosphoproteomic study was performed to identify key pathways activated by the LGG lysate. A Staphylococcus epidermidis lysate control was used to filter out any nonspecific bacterial effects. Immunostaining of nuclear proteins was used to visualize the effects of LGG lysate on lamin dispersal and nuclear degradation. short hairpin RNA filaggrin knockdown normal human keratinocytes were developed to create a 'barrier impaired' model to assess ability of the lysate to 'rescue' the skin barrier. Results The LGG lysate was shown to increase tight junction protein expression in normal human keratinocytes and increase keratinocyte terminal differentiation markers. We present Results that show decreased tight junction protein expression in filaggrin knockdown cells. Conclusions These Results show the potential for LGG lysate to enhance and restore skin barrier function and begin to suggest potential mechanisms for this effect. [ABSTRACT FROM AUTHOR]

Maifan stone is a kind of mineral medicine in Chinese medicine, which has good adsorption, dissolution, mineralization and biological activity. It has an excellent therapeutic effect on livestock, poultry and aquatic animals suffering from intestinal diseases. This study explored the effect of Maifan stone on the growth ability of Lacticaseibacillus rhamnosus GG (L. rhamnosus GG) and the effect of Maifan stone‐L. rhamnosus GG‐fermented product on the intestinal inflammation and gut microbiota. We find that Maifan stone can adsorb L. rhamnosus GG to form a carrier bacteria. Maifan stone has the characteristics of acid tolerance and bile salt tolerance and can also improve the activity of L. rhamnosus GG in artificial gastrointestinal juice. The fermented product can reduce the degree of diarrhoea and colon pathology in rats to a certain extent and significantly improve intestinal inflammatory factors and gut microbiota. This study improves the application effect of L. rhamnosus GG in the prevention and treatment of diarrhoea animals and provides a scientific basis for the rational development of Maifan stone resources. [ABSTRACT FROM AUTHOR]

Ionizing radiation is widely applied in food production as preservation technology and for correction of the gut microbiome of cancer patients, rescuers, astronauts etc. Lactic acid bacteria (LAB) can be used for the same reason. The main goal of this study was to investigate the effect of irradiation on some activities of Lactobacillus rhamnosus MDC 9661 and its effect on the survival of irradiated rats. The results indicate that both ultraviolet (during 45 min) and X‐ray irradiations (with 2 Sv) decreased the CFU and the antibacterial activity of the strain. Higher than 700 Sv dose of X‐ray irradiation resulted in the total inhibition of antibacterial activity with the total reduction of colony forming units less than 10 cells ml−1, while irradiated with 1000 Sv dose L. rhamnosus MDC 9661 did not lose its proteolytic activity. It was also shown that L. rhamnosus MDC 9661 was not immunogenic in the organism of the rats and cannot lead to the development of autoimmune responses. L. rhamnosus MDC 9661 demonstrated the necessary properties for probiotics and can be effectively used for the correction of the gut microbiome of all target groups. The co‐aggregation of the cells is one of the mechanisms for resistance of LAB to irradiation. [ABSTRACT FROM AUTHOR]

Background Although pathogenic gut microbiota causes gut leakage, increases translocation of uremic toxins into circulation and accelerates CKD progression, the local strain of Lactobacillus rhamnosus L34 might attenuate gut leakage. We explored the effects of L34 on kidney fibrosis and levels of gut-derived uremic toxins (GDUTs) in 5/6 nephrectomy (5/6Nx) mice. Methods At 6 weeks post-5/6Nx in mice, either L34 (1 × 106 CFU) or phosphate buffer solution (as 5/6Nx control) was fed daily for 14 weeks. In vitro , the effects of L34-conditioned media with or without indoxyl sulfate (a representative GDUT) on inflammation and cell integrity (transepithelial electrical resistance; TEER) were assessed in Caco-2 (enterocytes). In parallel, the effects on proinflammatory cytokines and collagen expression were assessed in HK2 proximal tubular cells. Results At 20 weeks post-5/6Nx, L34-treated mice showed significantly fewer renal injuries, as evaluated by (i) kidney fibrosis area (P < 0.01) with lower serum creatinine and proteinuria, (ii) GDUT including trimethylamine- N -oxide (TMAO) (P = 0.02) and indoxyl sulfate (P < 0.01) and (iii) endotoxin (P = 0.03) and serum TNF-α (P = 0.01) than 5/6Nx controls. Fecal microbiome analysis revealed an increased proportion of Bacteroidetes in 5/6Nx controls. After incubation with indoxyl sulfate, Caco-2 enterocytes had higher interleukin-8 and nuclear factor κB expression and lower TEER values, and HK2 cells demonstrated higher gene expression of TNF-α, IL-6 and collagen (types III and IV). These indoxyl sulfate–activated parameters were attenuated with L34-conditioned media, indicating the protective role of L34 in enterocyte integrity and renal fibrogenesis. Conclusion L34 attenuated uremia-induced systemic inflammation by reducing GDUTs and gut leakage that provided renoprotective effects in CKD. [ABSTRACT FROM AUTHOR]

This proof-of-principle study aims to find commensal oral bacteria that can produce extracellular polymeric substances (EPS), which have similar lubrication properties to saliva and could serve as saliva substitutes. Saliva and plaque samples were collected from 21 generally healthy individuals. Primary screening was done by conventional culturing and Gram-staining; all species selected for further analysis were identified by MALDI-TOF and deposited in DSMZ. Lactobacillus gasseri (DSM32453 and DSM32455), Lactobacillus rhamnosus (DSM32452), Lactobacillus paracasei (DSM32454), and Streptococcus sanguinis (DSM32456) produced 413.6, 415.7, 431.1, 426.8, and 877.6 µg/ml of EPS, respectively. At the same time calcium dissolution could not be detected for both L. gasseri strains, minimal dissolution for the other three: S. sanguinis 0.3 mm, and 3.7 mm for L. rhamnosus and L. paracasei. There were no differences found between the EPS samples and the saliva for the effect of shear rate on the viscosity and for the effect of sliding speed on lubrication properties. In conclusion, five commensal bacterial strains have been isolated, all able to produce EPS and lead to no or to low calcium dissolution. EPS produced exhibits rheological and tribological properties comparable to human saliva. A total of four out of five selected strains are probiotic and, therefore, may exhibit additional beneficial influence within the oral cavity. [ABSTRACT FROM AUTHOR]

Aim: Dietary intervention using probiotic bacteria has emerged as a promising preventive strategy in addressing foodborne infections or gastrointestinal disorders. This study investigated the immunomodulatory effects of Lactobacillus fermentum (MTCC‐5898) on Escherichia coli‐induced inflammatory responses in intestinal epithelial cells. Methods and Results: The immune response of intestinal cells (Caco‐2) in the presence of probiotic Lact. fermentum was determined during exclusion, competition and displacement of E. coli as the inflammatory agent. To achieve this objective, transcriptional modulation of key immune‐related genes (cytokines, pattern recognition receptors and NF‐κB), release of cytokines and nuclear translocation of the NF‐κB subunit p‐65 were studied. Expression of pro‐inflammatory cytokines IL‐8, TNF‐α, IFN‐ϒ and IL‐23 was high in E. coli‐exposed intestinal cells. However, overexpression of these E. coli‐induced pro‐inflammatory cytokines was prevented by Lact. fermentum during exclusion and competition assays. It also modulated the transcriptional expression of regulatory cytokines (IL‐10 and TGF‐β), pattern recognition receptors (TLR‐2 and TLR‐4) and genes associated with master inflammatory regulators (NF‐κB and SIGIRR) to reduce E. coli‐induced inflammation. The protective effect of Lact. fermentum was further confirmed by suppression of nuclear translocation of cytoplasmic NF‐κB subunit (p‐65). Conclusion: Lactobacillus fermentum alleviated E. coli‐induced inflammatory responses by modulating the NF‐κB signalling besides pro‐inflammatory and regulatory cytokines expression. Significance and Impact of the Study: Lactobacillus fermentum holds significant promise as a potent nutraceutical that prevents and manages inflammatory gut‐associated dysfunctions. [ABSTRACT FROM AUTHOR]

This work compared response surface methodology (RSM) and genetic algorithm (GA) analysis to optimize the lactic acid content by Lactobacillus rhamnosus PTCC 1637 and Lactobacillus acidophilus PTCC 1643 in a medium based on date syrup. Three parameters including concentrations of sucrose [10 and 20% (w/w)] and yeast extract [1, 2 and 3% (w/w)] along with different amplitudes of ultrasound (30 kHz, 25 and 50%) were investigated in terms of their impacts on both viable cell counts and lactic acid production. Regarding RSM and GA, optimized samples were selected by achieving high lactic acid concentration. The results indicated that an increase in the amounts of sucrose and yeast extract led to increasing the cell growth and lactic acid production. Application of ultrasound at 25% amplitude significantly (P  < 0.05) stimulated the fermentation process. However, increasing the amplitude to 50% significantly (P  < 0.05) decreased the lactic acid production compared with the control samples. The best treatment was observed at 20% sucrose, 3% yeast extract and 25% ultrasound amplitude. The present results indicate that the best productivity of lactic acid can be achieved at appropriate fermentation conditions, including a suitable amplitude of ultrasound and supplementation of date syrup. [ABSTRACT FROM AUTHOR]

Probiotics are considered as a potential nonantibiotic feed additive that may improve intestinal health of pigs. However, there is limited information on potential mechanism of probiotic Lactobacillus rhamnosus. The study was carried out to evaluate effects of probiotic Lactobacillus rhamnosus on intestinal morphology, blood profiles, immune responses, and ileal gene expression of weaned pigs. In a randomized complete block design [block: initial body weight (BW)], a total of 100 weaned pigs (7.99 ± 0.85 kg of average initial BW; 28 days of age) were assigned to two dietary treatments (5 pigs/pen; 10 replicates/treatment). Dietary treatments were fed for 4 weeks: 1) a commercial basal nursery diet (CON), and 2) CON + 0.02% probiotic Lactobacillus rhamnosus IDCC 3201 (PRO). Probiotic Lactobacillus rhamnosus was obtained from a commercial company (ILDONG Bioscience Co., Ltd., Pyeongtaek, Republic of Korea). Blood was collected from randomly selected one pig per pen on day 1, 7, 14, and 28 for blood profiles and immune responses. On the final day of study, pigs were euthanized to collect each part of small intestine for measuring intestinal morphology and ileal gene expression. Measurements were 1) villus height, width, and area, crypt depth, villus height to crypt depth ratio (VH:CD), and number of goblet cells by the hematoxylin and eosin staining method, 2) number of white blood cells (WBC), red blood cells (RBC), and hematocrit (HCT) by an automated hematology analyzer calibrated for porcine blood, 3) immunoglobulin G (IgG), IgA, tumor necrosis factor-α (TNF-α), transforming growth factor-β1 (TGF-β1), interleukin-10 (IL-10), and C-reactive protein in serum by the ELISA kits, and 4) ileal gene expression of tight junction-related proteins and inflammatory cytokines by qRT-PCR. Data were analyzed using PROC GLM of SAS. Experimental unit was the pen. Statistical model for intestinal morphology, blood profiles, and immune responses included dietary treatment as a main effect and initial BW as a covariate. The t-test was used for ileal gene expression. The PRO increased number of goblet cells in duodenum (P < 0.05) and VH:CD in ileum (P < 0.10) compared with CON. Pigs fed PRO had decreased HCT on day 7 (P < 0.10), WBC on day 28 (P < 0.05), and serum IgA concentrations on day 28 (P < 0.05) than those fed CON, but RBC on day 7 and 14 were greater (P < 0.05). The PRO upregulated (P < 0.05) claudin-3, occludin, tight junction protein-1 genes in ileum compared with CON, but TNF-α, interferon-α, IL-1α, IL-1β, IL-6, and IL-8 genes were downregulated (P < 0.05). In conclusion, addition of probiotic Lactobacillus rhamnosus in the nursery diet improved gut health via enhancing intestinal morphology and upregulating ileal tight junction-related protein genes, and modified blood parameters and ileal inflammatory cytokine genes. [ABSTRACT FROM AUTHOR]

Dietary probiotics were used in the animal feeds because of the beneficial effect of improving gut health and immune responses as one of the potential alternatives to antibiotics. This study was conducted to evaluate the effect of dietary supplementation of Lactobacillus rhamnosus (ILDONG Bioscience Co. Ltd., Pyeongtaek, Republic of Korea) on blood profiles, small intestinal morphology, immune responses, and gene expression of tight junction proteins and inflammatory cytokines in ileum of weaned pigs. A total of 40 weaned pigs [6.21 ± 0.55 kg of initial body weight (BW); 28-day old] were allotted to 2 dietary treatments (4 pigs/pen; 5 replicates/treatment) in a randomized complete block design (block: initial BW) and fed for 4 weeks. Dietary treatments were 1) a commercial basal nursery diet (CON), and 2) CON + 0.02% Lactobacillus rhamnosus IDCC 3201 (PRO). The blood samples were collected from randomly selected 1 pig per pen on day 1, 7, 14, and 28 for blood profiles and immune responses. On the last day of experiment, the pigs were euthanized to collect duodenal, jejunal, and ileal segments for intestinal morphology and gene expression analysis. Intestinal morphology was measured villus height, width, area, crypt depth, villus height to crypt depth ratio, and number of goblet cells by hematoxylin and eosin staining method. The blood profiles were measured by using an automated hematology analyzer. The serum samples were used to measure concentrations of cytokines and cortisol by using porcine immunoassay ELISA kits. The gene expression of tight junction proteins and cytokines were measured by qRT-PCR. Data were analyzed by PROC GLM procedure of SAS. Experimental unit was a pen. Statistical model for blood profiles, immune responses, and intestinal morphology included dietary treatment as a main effect and initial BW as a covariate. The t-test was used for ileal gene expression. Pigs fed PRO had lower (P < 0.05) platelet level, crypt depth in duodenum and ileum, serum tumor necrosis factor-α (TNF-α), and interleukin-6 than those fed CON. The pigs fed PRO downregulated (P < 0.05) TNF-α and interleukin-8 genes compared with those fed CON, but upregulated (P < 0.05) claudin-1 and claudin-2 genes. In conclusion, probiotic Lactobacillus rhamnosus supplementation in nursery diet improved gut health through modulating blood profiles and immune responses, enhancing intestinal morphology, and regulating tight junction protein genes expression in ileum of weaned pigs. Keywords: Lactobacillus rhamnosus, weaned pigsdownregulated (P < 0.05) TNF-a and interleukin-8 genes compared with those fed CON, but upregulated (P < 0.05) claudin-1 and claudin-2 genes. In conclusion, probiotic Lactobacillus rhamnosus supplementation in nursery diet improved gut health through modulating blood profiles and immune responses, enhancing intestinal morphology, and regulating tight junction protein genes expression in ileum of weaned pigs. [ABSTRACT FROM AUTHOR]

The swine industry has to find ways to make diets more efficient in terms of growth performance and swine intestinal health. Thus, this study was carried out to verify the effect of dietary supplementation of the probiotic, Lactobacillus rhamnosus, on growth performance, frequency of diarrhea, nutrient digestibility, and blood biochemical parameters of weaned pigs. A total of 40 weaned pigs [6.21 ± 0.55 kg of initial body weight (BW)] were assigned to 2 dietary treatments (4 pigs/pen; 5 replicates/treatment) in a randomized complete block design (block: initial BW). This experiment was performed for 4 weeks. Dietary treatments were a commercial basal nursery diet (CON) and CON supplemented with 0.02% Lactobacillus rhamnosus IDCC 3201 (PRO; ILDONG Bioscience Co., Ltd., Pyeongtaek, Republic of Korea). Average daily gain (ADG), average daily feed intake, and gain to feed ratio (G:F) were measured on day 1, 7, 14, and 28. The frequency of diarrhea was measured for the first 2 weeks after weaning. Diarrhea score of each pen was visually estimated from 1 to 5 (1 = normal feces, 2 = moist feces, 3 = mild diarrhea, 4 = severe diarrhea, and 5 = watery diarrhea). For the last week of experimental period, pigs were fed respective dietary treatments containing 0.2% chromium oxide. Fecal samples from randomly selected one pig per pen were collected daily by rectal palpation for the last 3 days. The samples from the diet and feces of each treatment were used to measure the apparent total tract digestibility of dry matter (DM), energy (EN), and crude protein (CP). Serum samples were collected for blood biochemical parameters. Growth performance, nutrient digestibility, and blood biochemical parameters were analyzed by the General Linear Model of SAS. The frequency of diarrhea was analyzed by the Chi-square test. The experimental unit was a pen. Statistical model for growth performance, nutrient digestibility, and biochemical parameters included dietary treatment as a main effect and initial BW as a covariate. Pigs fed PRO had greater (P < 0.05) ADG and G:F than those fed CON from day 1 to 7, and The PRO had greater (P < 0.05) ADG than CON from day 15 to 28. The PRO increased ADG during overall the experimental period (P < 0.10), nutrient digestibility of DM, EN, and CP (P < 0.05) compared with CON. However, no differences were found on frequency of diarrhea and blood biochemical parameters between dietary treatments. In conclusion, the dietary supplementation of Lactobacillus rhamnosus improved growth performance and nutrient digestibility. [ABSTRACT FROM AUTHOR]

Probiotics are microorganisms which have positive effects on human and animal gut health, and generally recognized as safe ingredients. This study was conducted to investigate the effects of probiotic Lactobacillus rhamnosus (LR) on growth performance, diarrhea frequency, and blood biochemical parameters of weaned pigs. A total of 100 weaned pigs [(Landrace × Yorkshire) × Duroc; 7.99 ± 0.85 kg of initial body weight (BW); 28 days old] were used in a 28-day trial. All pigs were randomly assigned to 2 dietary treatments in a randomized complete block design (block: initial BW) and there were 10 replication pens in each treatment with 5 pigs/pen. Dietary treatments were as follows: 1) a commercial weaner diet based on cornsoybean meal (CON), and 2) CON + 0.02% probiotic LR IDCC 3201 (PRO). All pigs were housed in an environmental controlled room and had free access to feed and water throughout the experimental period. Probiotic LR was obtained from commercial company (ILDONG Bioscience Co., Ltd., Pyeongtaek, Republic of Korea). Pigs BW was individually measured on day 1, 7, 14, and 28, and feed consumption was recorded on a pen basis on the same day to calculate average daily gain (ADG), average daily feed intake (ADFI), and gain to feed ratio (G:F). Diarrhea frequency was recorded by visual observation for 2 weeks after weaning and calculated as a percentage. Blood samples for biochemical parameters were collected from randomly selected one pig per pen on day 28 and analyzed by specific kit and clinical auto analyzer. Analysis items were total protein, aspartate transaminase, alanine transaminase, blood urea nitrogen, calcium (Ca), inorganic phosphorus, magnesium, total cholesterol, triglyceride, glucose, albumin, and creatinine. Data were analyzed using PROC GLM of SAS. Experimental unit was a pen. Statistical model for every measurement included dietary treatment as a main effect and initial BW as a covariate. The Chi-square test was used for diarrhea frequency. During day 1 to 7, 8 to 14, 1 to 14 and overall experiment date, ADG of pigs that fed PRO was greater (P < 0.05) than those fed CON. Pigs fed PRO had increased (P < 0.05) G:F during day 1 to 7 and 1 to 14 compared with those fed CON. Pigs fed PRO tended to have less (P < 0.10) diarrhea frequency during the first week of experiment than those fed CON. On blood biochemical parameters, pigs that fed PRO had greater (P < 0.05) concentrations of Ca (mg/dL) on day 28 than those fed CON. In conclusion, dietary supplementation of probiotic LR in weaner diet improved growth performance, reduced diarrhea frequency, and modulated blood biochemical parameters. [ABSTRACT FROM AUTHOR]

Aims: The catabolism products of the fermentation process of selected hydroxycinnamic acids initiated by different species of Lactobacillus strains were identified. Methods and Results: Three dietary supplements (Sanprobi IBS®, BioGaia ProTectis Baby® and Dicoflor 60®) were used to isolate the Lactobacillus strains. The overnight bacterial cultures (18 h) were diluted and grown in a microaerophilic atmosphere at 37°C. Then, each phenolic acid was added to bacterial cultures and incubated for 24 h at 37°C. Samples were collected at specific intervals for a further 24 h of incubation. LC‐MS/MS was used for the identification of metabolism products of selected phenolic acids. Conclusions: The phenolic acids were resistant to the Lactobacillus rhamnosus GG. Lactobacillus plantarum 299v caused degradation of caffeic and ferulic acids. The former was degraded either to dihydrocaffeic acid or to 4‐vinylcatechol and 4‐ethylcatechol. Ferulic acid was degraded only to dihydroferulic acid. Lactobacillus reuteriDSM 17938 caused only the degradation of chlorogenic acid (5‐caffeoylquinic acid, referred to IUPAC nomenclature) to caffeic acid. Significance and Impact of the Study: Using of Lactobacilli as food additive should be taken into account that phenolic acids metabolism rate depends on not only the specific bacterial strain but also the structural properties of the acid. [ABSTRACT FROM AUTHOR]

The present study aimed to address the capability of the probiotic bacterium Lactobacillus rhamnosus IMC 501® to survive in seawater and the ability of Artemia metanauplii to incorporate it, as well as to analyse the potential effect of the probiotic as a control agent for potentially pathogenic Vibrionaceae bacteria in Artemia. The results demonstrate the ability of L. rhamnosus IMC 501® to survive in seawater for up to 30 h. They also advocate their capability to be efficiently incorporated into Artemia metanauplii at concentrations of 104 CFU per Artemia after 30 min of suspension in probiotic solution, thereby promoting a 1‐log reduction in Vibrionaceae levels after 3 h. These low levels of Vibrio bacteria were maintained for about 30 min after transfer into clear seawater, a sufficient time for Artemia to be ingested by aquatic organisms. These results contribute to broaden the knowledge on the suitability of probiotics as sustainable alternatives for the prevention/reduction of diseases in aquaculture facilities. [ABSTRACT FROM AUTHOR]

Artisanal products support the conservation of the indigenous biodiversity of food microbiomes, although they do not always comply to quality and hygienic requirements for the dairy industry. This study describes the development of an autochthonous starter culture to produce Matsoni, a traditional Georgian fermented milk. To this end, strains of lactic acid bacteria isolated from artisanal Matsoni samples were used to design a starter formulation reproducing the dominant microbial diversity, also preserving quality characteristics and ensuring the safety of the product. As a result, strains that represent the acidifying portion of the starter (Lactobacillus delbrueckii subsp. lactis , L. delbrueckii subsp. bulgaricus and Streptococcus thermophilus) were combined in different ratios and strain combinations, together with cultures of Lactobacillus rhamnosus that were chosen for their potential beneficial traits. The strain association acting better in milk cultures at laboratory scale was selected as starter culture for the production of Matsoni in pilot-scale industrial trials. [ABSTRACT FROM AUTHOR]

The CoMiniGut in vitro model mimicking the small intestine of piglets was used to evaluate four probiotic strains for their potential as a preventive measure against development of diarrhea in weaned pigs. In the in vitro system, piglet digesta was inoculated with pathogenic enterotoxigenic Escherichia coli F4 (ETEC F4), and the short-chain fatty acid profile and the gut microbiota composition were assessed. A total of four probiotic strains were evaluated: Enterococcus faecium (CHCC 10669) , Lactobacillus rhamnosus (CHCC 11994) , Bifidobacterium breve (CHCC 15268) and Faecalibacterium prausnitzii (CHCC 28556). The significant differences observed in metabolite concetration and bacterial enumeration were attributed to variation in inoculating material or pathogen challenge rather than probiotic treatment. Probiotic administration influenced the microbiota composition to a small extend. Learnings from the present study indicate that the experimental setup, including incubation time and choice of inoculating material, should be chosen with care. [ABSTRACT FROM AUTHOR]

Summary: Alterations in the composition and reduced diversity of the infant microbiome are associated with allergic disease in children. Further, an altered microbiota is linked to immune dysregulation, including skewing of different T helper (Th) subsets, which is also seen in atopic individuals. The aim of this study was, therefore, to investigate the associations between gut lactobacilli and Th‐related plasma factors in allergy development during childhood. A total of 194 children with known allergy status at 1 year of age were followed to 10 years of age. We used real‐time polymerase chain reaction (PCR) to investigate the presence of three lactobacilli species (Lactobacillus casei, L. paracasei, L. rhamnosus) in infant fecal samples (collected between 1 week and 2 months of age) from a subgroup of children. Plasma chemokines and cytokines were quantified at 6 months and at 1, 2, 5 and 10 years of age with Luminex or enzyme‐linked immunosorbent assay (ELISA). Fractional exhaled nitrogen oxide (FeNO) was measured and spirometry performed at 10 years of age. The data were analysed by non‐parametric testing and a logistic regression model adjusted for parental allergy. An absence of these lactobacilli and higher levels of the chemokines BCA‐1/CXCL13, CCL17/TARC, MIP‐3α/CCL20 and MDC/CCL22 in plasma at 6 months of age preceded allergy development. The presence of lactobacilli associated with lower levels of atopy‐related chemokines during infancy, together with higher levels of interferon (IFN)‐γ and lower FeNO during later childhood. The results indicate that the presence of certain lactobacilli species in the infant gut may influence allergy‐related parameters in the peripheral immune system, and thereby contribute to allergy protection. [ABSTRACT FROM AUTHOR]

Summary: This review is part of a series of annual updates that summarize the evidence base for atopic eczema (AE). The aim is to provide a succinct guide for clinicians on the key findings from 14 systematic reviews on the prevention and topical treatment of AE published or indexed in 2018. Various supplements, including long‐chain polyunsaturated fatty acids, vitamin D and the probiotic Lactobacillus rhamnosus GG, given prenatally and postnatally, have not been shown to prevent AE in infants, although mixed strains of probiotics may decrease the risk of AE if given to the mother during pregnancy and to the infant for the first 6 months of life. In the postnatal period, there is no evidence that hydrolysed formula, compared with cow's milk formula (CMF), reduces the risk of AE in partially breastfed infants. However, weak evidence suggests that a specific partially hydrolysed whey formula decreases the risk of AE compared with CMF. No specific skin practices can be recommended to reduce the eczema risk in healthy term babies. There is weak evidence of a low risk of reversible hypothalamic–pituitary–adrenal axis suppression following 2–4 weeks of treatment with low‐potency topical steroids, and conflicting evidence as to whether bleach bathing affects skin flora or AE severity. A single study demonstrated that the topical Janus kinase inhibitor tofacitinib at 2% significantly reduces the Eczema Area and Severity Index compared with vehicle. Topical naltrexone cream 1% improves pruritus (measured using a visual analogue scale) by 30% more than placebo. There is weak evidence that topical alternative therapies, including antioxidants, micronutrients and some herbal medicines, may improve AE. Click here for the corresponding questions to this CME article. [ABSTRACT FROM AUTHOR]

Background Strain Lactobacillus rhamnosus GG is one of the best-studied and most widely used probiotic strains, with various health benefits. Because probiotic health benefits and safety are strain specific, the availability of a reliable assay for specific identification of Lactobacillus rhamnosus GG is vital to ensure probiotic efficacy. Objective To design and validate a probe-based real-time PCR assay for specific identification of strain Lactobacillus rhamnosus GG. Method Rapid Annotation using Subsystem Technology (RAST) was used to find a unique sequence region in the genome of Lactobacillus rhamnosus GG. A probe-based assay was designed and evaluated for specificity, sensitivity, efficiency, repeatability, and reproducibility. Results RAST identified a unique gene coding for a hypothetical protein in the genome of Lactobacillus rhamnosus GG. The assay successfully amplified all 22 target samples and did not amplify any of the 28 non-target strains, achieving 100% true positive and 0% false positive results. The Limit of Detection (LOD) was determined to be 0.001 ng. Reaction efficiency values, from three dilution series, were 96.4%, 93.3%, and 96.8% with R square values of 0.9974, 0.9981, and 0.9998, respectively. Relative standard deviation (RSD, %) of repeatability was below 1% and RSD of reproducibility was below 4%. Conclusions This Lactobacillus rhamnosus GG specific assay proved to be specific, sensitive, efficient, and reproducible. Since the assay was evaluated on two real-time PCR platforms, including a portable one, the assay can be used for onsite testing throughout the supply chain. Highlights The availability of validated and reliable assays for strain-specific identification plays a vital role in achieving compliance in probiotic products. [ABSTRACT FROM AUTHOR]

The objective was to carry out cytotoxicity assays in Vero cells and cytokines analyses in Balb/c mice as safety assessments to evaluate the probiotic mixture (M) Saccharomyces cerevisiae RC016 (Sc) and Lactobacillus rhamnosus RC007 (Lr) for use as feed additive. Vero cells (104 cells per well) were exposed to Sc (2·08 × 107, 2·08 × 106; 2·08 × 105 cells per ml), Lr (8·33 × 107; 8·33 × 106; 8·33 × 105 cells per ml) and their M (1 : 1). Sc concentrations did not affect the Vero cells viability; in contrast, they were lower when exposed to Lr (P ˂ 0·0001). Vero cells showed increasing viability with M decreasing concentrations (91% viability with M2). Control BALB/c mice received only phosphate buffer saline and the others received the M. The IL‐10, IL‐6 and TNFα concentrations from intestinal fluid were analysed and no significant differences were observed among treatments. The same occurred with the ratio between IL‐10/TNF‐α. Beneficial effects of probiotics are associated with the regulation of the excessive inflammatory response; it is desirable they can modulate the cytokines production only under pathological conditions. Here, M administration to healthy mice did not induce negative side effects and expands the knowledge about beneficial effects of using probiotic microorganisms in mixture for feed additives development. [ABSTRACT FROM AUTHOR]

Aim: This study investigated the in vitro mechanism of action of a commercial citrus EO, Brazilian orange terpenes (BOT), on an enterotoxigenic Escherichia coli (ETEC) isolated from pig gut and on Lactobacillus rhamnosus. Methods and Results: Firstly, bacteria were exposed sequentially to BOT every 3 h (three times) at sub‐minimal inhibitory concentrations and results showed that sequential exposure to BOT provoked a higher reduction of bacteria viability than a single exposure and the reduction of ETEC viability was higher compared to that of L. rhamnosus. Then, evaluation of the BOT effects on the cell membrane permeability and integrity, indicated that BOT increased the membrane permeability and caused disruptive effects on the integrity of bacterial cells as reflected by an increase of the relative electric conductivity and the release of essential cell constituents. Interestingly, BOT effects were more pronounced on the ETEC than on L. rhamnosus. This was ratified by scanning electron microscopy, which showed more noticeable morphological damages and disturbances on ETEC cells than on the L. rhamnosus cells. Limonene was detected as the major compound in BOT by polar/nonpolar GC‐MS (78·65%/79·38%). Conclusions: Results revealed that the probable mechanism of the selective antibacterial action of the citrus EO, BOT, can be described as altering more remarkable the permeability and integrity of the cytoplasmic membrane as well as the external structure of ETEC cells than L. rhamnosus cells. Significance and Impact of the Study: This study provides information about the mechanism of antibacterial action displayed by a citrus EO, a by‐product of the citrus processing industry, as a natural alternative to antibiotics used in pig production sector to combat pathogens such as ETECs. [ABSTRACT FROM AUTHOR]

Aim: To effectively make high‐quality silage in hot and humid area. Methods and Results: The natural lactic acid bacteria (LAB) strains CZ149, XH358, XH753 and XH761 isolated from corn and Napier grass were screened by the potential of low pH growth and high lactic acid production, and their effect on silage quality, aerobic stability and aflatoxin B1 production of whole‐crop corn was also studied in Sichuan, China. Four selected strains were Gram‐positive and catalase‐negative, rod‐shaped strains that are able to grow at pH 3·5 and at 45°C. Strains CZ149, XH358, XH753 and XH761 were identified as Lactobacillus plantarum, L. salivarius, L. rhamnosus and L. paracasei, respectively. After 60 days of fermentation, all LAB strains showed no significant relationship with the quality of corn silage, whereas the lowest aflatoxin B1 and lactic‐to‐acetic ratio were detected in strain XH753‐treated silage. Strain CZ149‐treated silage showed worse aerobic stability and higher aflatoxin B1 concentration than control, whereas strain XH753‐treated silage had better aerobic stability and lowest aflatoxin B1 concentration after aerobic exposure in hot and humid condition for 5 days. Conclusions: The three L. plantarum strains used in this study are not suitable as inoculants for local whole‐crop corn silage, whereas L. rhamnosus 753 could prolong the aerobic stability and inhibit the accumulation of aflatoxin B1 at hot and humid condition. Significance and Impact of the Study: This study provides new information of LAB inoculants for corn silage in hot and humid areas. Especially, a screened strain of Lactobacillus rhamnosus 753 can be used as a candidate strains to make high‐quality silage in tropical and subtropical areas. [ABSTRACT FROM AUTHOR]

Aim: Studies on the anticancer effects of synbiotic fermented soymilk are rare. The aim of the present study was to evaluate the effect of synbiotic fermented soymilk supplemented with xylooligosaccharides and inoculated with Weissella cibaria FB069 (FSMXW) in the proliferation of colon cancer cell and compare it with the effect of soymilk inoculated with Lactobacillus rhamnosus GG (LGG). Methods and Results: Both FB069 and LGG were able to grow in soy‐based products and rapidly reduce their pH as a result of fermentation. The addition of XOS significantly enhanced the acidification rate, viscosity and total cell concentration in fermented soymilk inoculated with W. cibaria FB069. However, the same effect was not observed following inoculation with LGG. Moreover, the synbiotic FSMXW showed higher dextran, folate, GABA and aglycone content. FSMXW inhibited the proliferation of Caco‐2 and HCT116 cell lines, by reducing the transcription of MD2, TLR4, MyD88, and NF‐κb. Conclusions: The synbiotic soymilk containing XOS and W. cibaria FB069 increase nutrient and functional compounds through fermentation process. Significance and Impact of the Study: Our finding suggests that W. cibaria and XOS can be potentially employed in developing functional foods and health‐related products. [ABSTRACT FROM AUTHOR]

Probiotics can stabilize gut flora, regulate intestinal immunity and protect the host from enteric diseases; however, their roles in oral health have received little attention compared to their roles in gut health. Nowadays, the prevalence of sugar‐sweetened foods and abuse of antibiotics contribute towards dysbiosis of oral microbiota and drug resistance development in oral pathogens, resulting in various intractable oral diseases. We screened the antibacterial activities of viable and heat‐killed probiotic strains against the oral pathogens Streptococcus mutans, Porphyromonas gingivalis, Fusobacterium nucleatum and Aggregatibacter actinomycetemcomitans. The probiotic strains Lactobacillus salivarius subsp. salicinius AP‐32, L. rhamnosus CT‐53, L. paracasei ET‐66 and Bifidobacterium animalis subsp. lactis CP‐9 displayed strong antipathogenic activities, whereas heat‐killed AP‐32, CT‐53 and ET‐66 displayed high levels of pathogen inhibition. The antibacterial activities of these probiotics were not associated with their H2O2 production; L. acidophilus TYCA02 produced high levels of H2O2 but merely exhibited moderate antibacterial activities. Oral tablets containing probiotics showed positive inhibitory effects against oral pathogens, particularly those containing viable probiotics. Our results indicate that probiotics prevent the growth of oral pathogens and improve oral health, providing insights into the antipathogenic efficacy of different probiotic species and their potential role in functional foods that improve oral health. Significance and Impact of the Study: Our study provides insights into the antipathogenic efficacy of different probiotic species and their potential roles in developing functional foods to improve oral health. We showed that the probiotic strains Lactobacillus salivarius subsp. salicinius AP‐32, L. rhamnosus CT‐53, L. paracasei ET‐66 and Bifidobacterium animalis subsp. lactis CP‐9 have great potential for use in the development of functional foods to improve oral health. Since active probiotics may provide strong and long‐term protection, the development of functional food products should favour the use of viable bacteria. [ABSTRACT FROM AUTHOR]

This study assessed whether antibiotics could alter gut microbiota to affect host growth and the possibility of alleviation by lactobacilli. We divided four-week-old BABL/c mice into control (Ctrl), antibiotic exposure (Abx), Lactobacillus plantarum PC-170 (PC), and Lactobacillus rhamnosus GG (LGG) group and the Abx, LGG, and PC group received an one-week antibiotic/antibiotic + probiotic treatment. The fecal microbiota and the expression of splenic cytokines were determined. Following the ceftriaxone treatment, the body weight gain of Abx was delayed compared with others. The ceftriaxone treatment significantly decreased the alpha-diversity of the fecal microbiota and altered the fecal microbiota but LGG and PC can partly alleviate the effect. At the end of the study, the microbial community of LGG and PC group were more similar to Ctrl compared with Abx group. The results indicated that ceftriaxone could significantly alter intestinal microbiota. Lactobacilli might alleviate the side effects of antibiotics by stabilizing the intestinal microbiota. Lactobacilli alleviate dysbiosis of the gut microbiota induced by antibiotic. [ABSTRACT FROM AUTHOR]

Aims: Probiotics have the ability to enhance the immune system, produce anti‐inflammatory action and promote wound healing process. The first aim of the study was to isolate pathogenic micro‐organisms from sites of chronic ulcerative lesion. The second aim was to evaluate probiotic efficacy of SYNBIO® (1:1 combination of Lactobacillus rhamnosus IMC 501® and Lactobacillus paracasei IMC 502®) in counteracting wound infections. Methods and Results: Several bacterial pathogens were isolated from chronic ulcerative lesions and identified by morphological, biochemical and molecular techniques. SYNBIO® probiotic formulation was investigated for its antimicrobial activity, minimum inhibitory concentration, co‐aggregation and adherence capacity against the isolated pathogens. Moreover, SYNBIO was also tested in combination with some medical devices, using an in vitro model, in order to simulate a real ulcerative wound infection. Probiotic formulation demonstrated an inhibitory action against all the tested pathogens and their mixture (MIX), with an increased ability of co‐aggregation during time. In addition, the adhesion percentage of probiotic micro‐organisms to human keratinocyte (HaCaT cells) and human fibroblasts (NHF), calculated by an in vitro model, was 19% and 17% respectively, highlighting the possibility to create a protective environment preventing pathogens' biofilm formation in order to contrast infections. Conclusions: SYNBIO® probiotics showed a very good antimicrobial capacity and adhesion percentage to HaCaT cells and fibroblasts, giving the opportunity to be successfully used as complement to conventional therapies in the treatment of chronic ulcerative lesions. Significance and Impact of the Study: A new therapeutic approach with probiotics (supplemented in topical applications, excluding side effects) able to eliminate pathogenic micro‐organisms and improve healing of chronic ulcerative lesions. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine whether Lactobacillus rhamnosus GG (LGG) components (surface layer protein, SLP; genomic DNA, gDNA; unmethylated cytosine‐phosphate‐guanine‐containing oligodeoxynucleotide, CpG‐ODN), alone or in combination, could affect immunomodulation, and evaluate the signalling mechanism in mouse macrophage RAW264.7 cells challenged with lipopolysaccharide (LPS). LGG components were used to treat cells before LPS stimulation. Cytokine and Toll‐like receptor (TLR) expression were assessed using real‐time quantitative PCR (RT‐qPCR). Mitogen‐activated protein kinase (MAPK), extracellular regulated protein kinase (ERK) and nuclear factor‐kappa B (NF‐κB) signalling pathways were evaluated using immunoblots and immunofluorescence. SLP or SLP + gDNA pre‐treatment significantly reduced the LPS‐induced mRNA expression of tumour necrosis factor alpha (TNF‐α). Pre‐treatment with LGG single components (SLP, gDNA or CpG) or their combinations (SLP + gDNA or SLP + CpG) significantly decreased the LPS‐induced interleukin‐6 (IL‐6) mRNA level (P < 0·05). Pre‐treatment with SLP or gDNA, alone or in combination, significantly suppressed LPS‐induced TLR2 and TLR4 mRNA levels (P < 0·05). SLP pre‐treatment also significantly decreased the LPS‐induced expression of TLR9 (P < 0·05). Pre‐treatment with LGG single components or combinations significantly suppressed the LPS‐induced phosphorylation levels of ERK (P > 0·05). In conclusion, pre‐incubation with LGG components, singly or in combination, generally inhibited the activation of TLR, MAPK and NF‐κB signalling pathways in LPS‐stimulated cells, leading to attenuated inflammatory cytokine TNF‐α and IL‐6 production. These results indicate that nonviable probiotic LGG components exert an anti‐inflammation effect on epithelial cells. Significance and Impact of the Study: Lactobacillus rhamnosus GG (LGG) is widely used as probiotics. However, its main components are not well known for affecting immunomodulation. This study investigated the effects of pre‐treatments with different components such as surface layer protein, genomic DNA and unmethylated cytosine‐phosphate‐guanine‐containing oligodeoxynucleotides, alone or in combination on immunomodulation, and evaluated the signalling mechanism in mouse macrophage RAW264.7 cells challenged with lipopolysaccharide. Pre‐incubation with components alone or in combination generally inhibited the activation of Toll‐like receptor, mitogen‐activated protein kinases, extracellular regulated protein kinases and nuclear factor‐kappa B signalling pathways in lipopolysaccharide‐stimulated cells, which generally leads to attenuated inflammatory cytokine interleukin‐6 and tumour necrosis factor alpha production. These results indicate that nonviable probiotic LGG components exert an anti‐inflammation effect on epithelial cells. [ABSTRACT FROM AUTHOR]

Probiotics are often used in critically ill patients to prevent antibiotic-associated complications, including Clostridium difficile colitis. However, clinical evidence of their efficacy is lacking. The objective of this study is to assess the impact of prophylactic probiotic administration on bowel function, gut microbial diversity, and nutritional markers in adult burn patients. A retrospective cohort study was done on 108 burn patients aged 18 to 89. Patients were given >1 million colony-forming units per day of Lactobacillus acidophilus and Lactobacillus rhamnosus. Testing for C. difficile was used as a surrogate marker for the presence of diarrhea. Serum C-reactive protein and prealbumin values were measured. Additionally, the gut microbial diversity of eight patients was tracked via 16S quantitative PCR before and throughout the course of a standard probiotic regimen. Patients receiving oral probiotics had more reported diarrhea in the first and second weeks of treatment. In the second week, C-reactive protein levels were increased, while serum prealbumin levels were lower in patients receiving probiotics, suggesting potential malabsorption. Additionally, there was no difference in C. difficile infection, sepsis rates, emesis, or gastric residuals, indicating an absence of therapeutic benefit for probiotic administration in burn patients. Furthermore, it was determined that no discernible benefit to gut microbial diversity was conferred by probiotic therapy. Prophylactic probiotics in burn patients are not associated with improvements in patient outcomes and may in fact be associated with an increased incidence of diarrhea and malabsorption. Additional research is needed before routine use in burn patients. [ABSTRACT FROM AUTHOR]

Probiotics including Lactobacillus and Bifidobacterium spp. have been known to control the balance of gut microbiota and are positively effective to host animals. In this study, 32 bacterial strains were isolated from Norwegian forest cat (castrated, 12 years old) and Persian cat (castrated, 10 years old) which were privately owned and had indoor access. Among them, Lactobacillus rhamnosus CACC612 (CACC612) and Bifidobacterium animalis subsp. Lactis CACC789 (CACC789) were selected as potential probiotics. From the characterization of the two strains, they showed equivalent acid tolerance, similar cell adhesion rate on HT-29 monolayer cell line, and superior bile salt tolerance, compared with Lactobacillus rhamnosus GG (LGG). Subsequently, they exhibited antibacterial activities against a broad spectrum of pathogenic bacteria such as E. coli KCTC 2617, Salmonella Derby NCCP 12238, Salmonella enteritidis NCCP 14546, Salmonella typhimurium NCCP 10328, Clostridium difficile JCM 1296T. From evaluating host effects, the viability of the feline macrophage cell line (Fcwf-4) increased with the treatment of CACC612 or CACC789 (P < 0.05). The induced expression of immune-related genes such as IFN-γ, IL1β, IL2, IL4, and TNF-α by immune stimulation was significantly attenuated by the treatment of CACC612 or CACC789 (P < 0.05). In addition, when 52 clinical factors of sera from 21 healthy cats were analyzed using partial least-squares discriminant analysis (PLS-DA), the animals were markedly clustered before and after CACC612 or CACC789 feeding. In addition, hemoglobin and Mean corpuscular hemoglobin concentration (MCHC) were significantly increased after CACC612 feeding (P < 0.05). Conclusively, these results could contribute to understanding the functional effects of feline-originated probiotics and supporting their industrial applications. [ABSTRACT FROM AUTHOR]

Aims: The aim of this study was to characterize Lactobacillus strains for their biological properties and amensalistic activities against genital and nongenital pathogens. Methods and Results: For the purpose, some special characteristics (H2O2, biofilm and antimicrobial substances production) as well as safety properties of 112 lactobacilli were evaluated. All the strains had good amensalistic characteristics, in particular cell‐free supernatants of 10 strains showed antibacterial activity against bacteria, as well as Candida sp. Moreover, these 10 strains were excellent biofilm producers. Conclusions: These results provide evidence for the possible use as probiotics for vaginal co‐therapy in case of dysbiosis. Significance and Impact of the Study: Recently, the problem of antibiotic resistance is constantly increasing, even though resources and energy are invested in order to increase knowledge on the mechanisms of action. Bacteriocins have a rapid mechanism of action, act at extremely low concentrations, are generally sensitive to proteases and they usually have a narrow killing spectrum; these characteristics reduce the possibility of the bacterium to develop resistance. This study is focused on the feasibility of a high production of antimicrobial substances and their characterization in order to be exploited as a therapeutic alterative or in co‐therapy with antibiotics in case of vaginal dysbiosis. [ABSTRACT FROM AUTHOR]

Pathogenic infection is one of the major causes of death in newborns. Antibiotic based therapies are still the major mode of treatment for infection. Increased usage of antibiotics leads to selective evolution of microorganisms and causes diseases in adulthood. Attempts to develop alternatives to antibiotics did not yield much success. A recent viable trend is to identify novel probiotics that could alleviate problems associated with over usage of antibiotics. We screened three different Lactobacillus species to establish their efficacy in neonates in protecting against Salmonella challenge. The methodologies employed are metagenomics, metabonomics, transcriptional profiling, molecular assays and behavioral studies. Among the three probiotics used, only Lactobacillus rhamnosus GG (LGG) treatment of the neonates resulted in rescuing of 80% of the Salmonella-infected mice. We have shown that LGG (MTCC #1408) can prevent Salmonella mediated infection in neonates. In the current report, results from histopathology, gene expression, neutrophil infiltration, metabolite and metataxonomic profiling, and protein level data suggested that LGG treatment of the neonates enhanced anti-inflammatory cytokine expression and increased the gut barrier function. The current report establishes the potential use of LGG in clinical intervention of infectious diseases. [ABSTRACT FROM AUTHOR]

Summary: A balanced microbiota of the gastrointestinal tract (GIT) is a prerequisite for a healthy host. The GIT microbiota in preterm infants is determined by the method of delivery and nutrition. Probiotics can improve the GIT microbiota balance and suitable animal models are required to verify their harmlessness. Preterm gnotobiotic piglets were colonized with Lactobacillus rhamnosus GG (LGG) to evaluate its safety and possible protective action against infection with an enteric pathogen, Salmonella Typhimurium (ST). Clinical signs (anorexia, somnolence, fever and diarrhea), bacterial interference and translocation, intestinal histopathology, transcriptions of claudin‐1, occludin and interferon (IFN)‐γ, intestinal and systemic protein levels of interleukin (IL)‐8, IL‐12/23 p40 and IFN‐γ were compared among (i) germ‐free, (ii) LGG‐colonized, (iii) ST‐infected and (iv) LGG‐colonized and subsequently ST‐infected piglets for 24 h. Both LGG and ST‐colonized the GIT; LGG translocated in some cases into mesenteric lymph nodes and the spleen but did not cause bacteremia and clinical changes. ST caused clinical signs of gastroenteritis, translocated into mesenteric lymph nodes, the spleen, liver and blood, increased claudin‐1 and IFN‐γ transcriptions, but decreased occludin transcription and increased local and systemic levels of IL‐8 and IL‐12/23 p40. Previous colonization with LGG reduced ST colonization in the jejunum and translocation into the liver, spleen and blood. It partially ameliorated histopathological changes in the intestine, reduced IL‐8 levels in the jejunum and plasma and IL‐12/23 p40 in the jejunum. The preterm gnotobiotic piglet model of the vulnerable preterm immunocompromised infant is useful to verify the safety of probiotics and evaluate their protective effect. We evaluated the effect of the previous colonization of the hysterectomy‐derived preterm gnotobiotic piglets with Lactobacillus rhamnosus GG against subsequent infection with Salmonella Typhimurium. This colonization with LGG partially ameliorated the deleterious effect of infection with S. Typhimurium, as it was evaluated by CFU enumeration, intestinal histology and the local and systemic values of IL‐8 and IL‐12/23 p40. It is the first use of preterm gnotobiotic piglets as an infectious model of vulnerable immunocompromised preterm infants to verify probiotic safety and its bacterial interference with an enteric pathogen. [ABSTRACT FROM AUTHOR]

Aims: (i) To determine the aflatoxin B1 (AFB1) adsorption and desorption dynamics in the presence of Lactobacillus rhamnosus RC007 under simulated transit of AFB1 at each gastrointestinal tract (GIT—saliva, stomach and intestine) stage consecutively and then, separately, (ii) to study the ability of L. rhamnosus RC007 to biotransform AFB1 as a strategy that complements the adsorption process. Methods and Results: The AFB1 adsorption and desorption assay simulating the GIT passage of AFB1 (93·89 ng g−1) in the presence of L. rhamnosus RC007 (108CFU per ml) was conducted. Moreover, lactic acid production was determined. Results demonstrated that predominant environmental conditions in salivary solution induced a low AFB1 adsorption, while the transit through the gastric solution and intestinal solution allowed high percentages of adsorption and did not generate significant AFB1 desorption. Conclusions: The AFB1 adsorption and desorption dynamics in the presence of L. rhamnosus RC007 was favoured by gastric and intestinal environment. Significance and Impact of the Study: The knowledge of the adsorption dynamics of AFB1 with a micro‐organism of interest will allow predicting its behaviour at each stage of the GIT. [ABSTRACT FROM AUTHOR]

Aims To evaluate the effects of spray drying of Lactobacillus rhamnosus 64 on its capacity to modulate the gut immune response and on the attenuation of TNBS-induced colitis in mice. Methods and Results Lactobacillus rhamnosus 64 was spray dried in cheese whey-starch solution and administered to mice for 3, 6 or 10 consecutive days. Peritoneal macrophage phagocytic activity, secretory IgA levels in the small intestinal fluid and TNF α, IFN γ, IL-10, IL-6 and IL-2 levels in homogenates of the small and large intestine were determined. The effects of spray drying were also evaluated in an acute model of Trinitrobenzenesulfonic acid ( TNBS)-induced colitis. A shift in the regulation of immune parameters, particularly the cytokine profile, was observed for mice treated with the spray-dried culture, compared to the profile observed in animals that received the strain as fresh culture (FC). The spray-dried culture of L. rhamnosus 64 showed anti-inflammatory properties in murine model of TNBS-induced colitis. Conclusions The spray-drying process of L. rhamnosus 64 in whey-starch modified its immunomodulating capacity in healthy animals and conferred enhanced protection in an in vivo model of inflammation. Significance and Impact of the Study Probiotic capacity can be affected by spray drying in relation to the properties observed for the strain as an overnight FC. This fact should be taken into account when producing the culture for its application in the industry. [ABSTRACT FROM AUTHOR]

Aim To develop a novel validated method for the isolation of Bifidobacterium animalis ssp. lactis BB-12 ( BB-12) from faecal specimens and apply it to studies of BB-12 and Lactobacillus rhamnosus GG ( LGG) recovered from the healthy human gastrointestinal ( GI) tract. Methods and Results A novel method for isolating and enumerating BB-12 was developed based on its morphologic features of growth on tetracycline-containing agar. The method identified BB-12 correctly from spiked stool close to 100% of the time as validated by PCR confirmation of identity, and resulted in 97-104% recovery of BB-12. The method was then applied in a study of the recovery of BB-12 and LGG from the GI tract of healthy humans consuming ProNutrients® Probiotic powder sachet containing BB-12 and LGG. Viable BB-12 and LGG were recovered from stool after 21 days of probiotic ingestion compared to baseline. In contrast, no organisms were recovered 21 days after baseline in the nonsupplemented control group. Conclusions We demonstrated recovery of viable BB-12, using a validated novel method specific for the isolation of BB-12, and LGG from the GI tract of healthy humans who consumed the probiotic supplement. Significance and Impact of the Study This method will enable more detailed and specific studies of BB-12 in probiotic supplements, including when in combination with LGG. [ABSTRACT FROM AUTHOR]

Food-related carbohydrates and proteins are often used as thermoprotectants for probiotic lactobacilli during industrial production and processing. However, the effect of inorganic salts is rarely reported. Magnesium is the second-most abundant cation in bacteria, and commonly found in various foods. Mg2+ homeostasis is important in Salmonella and has been reported to play a critical role in their thermotolerance. However, the role of Mg2+ in thermotolerance of other bacteria, in particular probiotic bacteria, still remains a hypothesis. In this study, the effect of Mg2+ on thermotolerance of probiotic lactobacilli was investigated in three well-documented probiotic strains, Lactobacillus rhamnosus GG, Lactobacillus casei Zhang and Lactobacillus plantarum P-8, in comparison with Zn2+ and Na+. Concentrations of Mg2+ between 10 and 50 mmol l−1 were found to increase the bacterial survival upon heat challenge. Remarkably, Mg2+ addition at 20 mmol l−1 led to a 100-fold higher survival of L. rhamnosus GG upon heat challenge. This preliminary study also showed that Mg2+ shortened the heat-induced extended lag time of bacteria, which indicated the improvement in bacterial recovery from thermal injury. Significance and Impact of the Study In order to improve the productivity and stability of live probiotics, extensive investigations have been carried out to improve thermotolerance of probiotics. However, most of these studies focused on the effects of carbohydrates, proteins or amino acids. The roles of inorganic salts in various food materials, which have rarely been reported, should be considered when incorporating probiotics into these foods. In this study, Mg2+ was found to play a significant role in the thermotolerance of probiotic lactobacilli. A novel strategy may be available in the near future by employing magnesium salts as protective agents of probiotics during manufacturing process. [ABSTRACT FROM AUTHOR]

Anchorless cell surface proteins (CSPs) were extracted with 1 M lithium chloride solution from Lactobacillus rhamnosus FSMM22. Loss of the anchorless CSPs resulted in a 2-fold decrease in FSMM22 cells bound to a constitutive extracellular matrix glycoprotein, laminin, in vitro. DNA-binding protein HU, glyceraldehyde-3-phosphate dehydrogenase, lactate dehydrogenase and 30S ribosomal protein S19 (RpsS) were identified by mass spectrometry in the extract as laminin-binding adhesins. Among the four proteins, RpsS was immunohistochemically confirmed to exist on the cell surface. Our findings strongly suggest that anchorless CSPs can enhance bacterial adhesion to the host. Laminin-binding cell surface proteins in Lactobacillus rhamnosus FSMM22. [ABSTRACT FROM AUTHOR]

Aims To investigate the in vivo effects of Lactobacillus rhamnosus GG ( LGG) on intestinal polyp development and the interaction between this single-organism probiotic and the gut microbiota therein. Methods and Results The ApcMin/+ mouse model was used to study the potential preventive effect of LGG on intestinal polyposis, while shotgun metagenomic sequencing was employed to characterize both taxonomic and functional changes within the gut microbial community. We found that the progression of intestinal polyps in the control group altered the community functional profile remarkably despite small variation in the taxonomic diversity. In comparison, the consumption of LGG helped maintain the overall functional potential and taxonomic profile in the resident microbes, thereby leading to a 25% decrease of total polyp counts. Furthermore, we found that LGG enriched those microbes or microbial activities related to short-chain fatty acid production (e.g. Roseburia and Coprococcus), as well as suppressed the ones that can lead to inflammation (e.g. Bilophila wadsworthia). Conclusions Our study using shotgun metagenomics highlights how single probiotic LGG may exert its beneficial effects and decrease polyp formation in mice by maintaining gut microbial functionality. Significance and Impact of the Study This probiotic intervention targeting microbiota may be used in conjugation with other dietary supplements or drugs as part of prevention strategies for early-stage colon cancer, after further clinical validations in human. [ABSTRACT FROM AUTHOR]

Aims This study evaluated the efficacy of a repeated oral treatment with two active pharmaceutical ingredients (Lcr Lenio® and Lcr Restituo®) derivated from the probiotic bacterial strain Lactobacillus rhamnosus Lcr35® in two animal models mimicking different features of irritable bowel syndrome ( IBS). IBS is characterized by visceral pain associated with alteration of bowel transit. IBS patients present visceral hypersensitivity with peripheral and central origins. Methods and Results The injection of 2,4,6-trinitrobenzenesulfonic acid ( TNBS) into the proximal colon as well as an acute partial restraint stress ( PRS) produces colonic hypersensitivity measured in conscious rats by a decrease in pain threshold in response to distal colonic distension. Visceral hypersensitivity was produced by injection of TNBS 7 days before colonic distension or by acute PRS on testing day. Treatments were performed once a day during eight consecutive days. Conclusions This study indicates that an 8-day probiotic treatment (Lcr Lenio and Lcr Restituo) produces an antihypersensitivity activity in both TNBS and PRS visceral pain models. As this probiotic strain attenuates peripherally and centrally induced visceral hypersensitivity in rats, it may be active in treatment of IBS symptoms. An immunomodulatory effect of the probiotics was highlighted in the TNBS model on the IL-23 secretion, suggesting a mechanism of action involving a regulation of the local IL-23/Th17 immune activation. Significance and Impact of the Study Two formulas of Lcr35® probiotic strain show very encouraging results for the treatment of IBS patients. Further studies are needed to better understand the role and mechanisms of probiotics on the pathogenesis of IBS. [ABSTRACT FROM AUTHOR]

Aim The aim of this study was to evaluate the potential anti- Candida effects of Lactobacillus rhamnosus ATCC 9595 on Candida albicans ATCC 18804 using in vitro and in vivo models. Methods and Results The in vitro analysis evaluated the effects of L. rhamnosus on C. albicans's biofilm formation by CFU count and metabolic activity, filamentation capacity, and adhesion ( ALS3 and HWP1) and transcriptional regulatory gene ( BCR1 and CPH1) expression. The in vitro results showed that both the L. rhamnosus cells and supernatant reduced C. albicans biofilm formation, filamentation and gene expression. In the in vivo study, the treatment with L. rhamnosus supernatant increased 80% the survival of Galleria mellonella larvae infected with C. albicans. Furthermore, the supernatant of L. rhamnosus recruited haemocytes into the haemolymph (2·1-fold increase). Conclusions Lactobacillus rhamnosus reduced the biofilm formation and filamentation of C. albicans in vitro by negatively regulating all studied C. albicans genes. Lactobacillus rhamnosus protected G. mellonella against experimental candidiasis in vivo. Significance and Impact of the Study This study is the first study to report the anti- Candida properties of L. rhamnosus ATCC 9595. The supernatant of this strain has immunomodulatory effects on the G. mellonella model and protects the larvae against pathogens. [ABSTRACT FROM AUTHOR]

Aims The study investigated whether the interaction with Lactobacillus rhamnosus (ATCC7469) interfere with the expression of virulence factors by Candida albicans (ATCC18804). Methods and Results These micro-organisms were grown in biofilms for 24, 48 and 72 h, Candida was isolated and the expression of the major virulence factors were investigated. The production of phospholipase, protease and haemolysin were observed in appropriate media; observation of germ tubes formation in serum; biofilm formation, after growth in microtitre plates and reading in spectrophotometer. Candida was also tested for antifungal sensitivity to amphotericin B, fluconazole and ketoconazole. The results were compared with the cells of Candida grown in the absence of lactobacilli (control group). Candida cells, which interacted with Lact. rhamnosus (test group), showed significantly lower proteinase and haemolysin activity, when compared with control group. The germ tube formation and biofilm formation capacity also decreased in tested groups, which demonstrated alterations in susceptibility to antifungal drugs. Conclusions The results suggest that Lact. rhamnosus is able to influence the expression of virulence factors by C. albicans and can alter its antifungal sensitivity profile. Significance and Impact of the Study These results suggest reduction in the pathogenicity of Candida and improvement in candidiasis therapy and control. [ABSTRACT FROM AUTHOR]

Aims Piliated Lactobacillus rhamnosus ( pLR) strains have attracted much attention owing to their excellent mucus adhering capacity and immunomodulatory effects. Here, we aimed to develop a rapid, sensitive method for isolating pLR strains in complex ecosystems using immunomagnetic separation ( IMS) with colony immunoblotting ( CIB). Methods and Results Magnetic nanobeads (diameter: 180 nm) conjugated with anti- pLR SpaA pilin antibodies (anti-SpaA) were prepared and used to preconcentrate pLR strains in samples, followed by confirmation with anti-SpaA-based CIB analysis. Under optimized experimental conditions, IMS- CIB selectively recovered pLR strains from 107 CFU ml−1 of faecal microbiota samples spiked with 2·9 × 101 to 2·4 × 106 CFU ml−1 of pLR strains. No positive colonies were detected in samples without addition of pLR strains. The detection limit of IMS- CIB was 29 CFU pLR ml−1 of faecal microbiota, which is much lower than that of CIB without IMS preconcentration (2·0 × 104 CFU ml−1). Conclusions IMS- CIB allowed selective preconcentration of pLR strains in highly heterogeneous bacterial suspensions and direct detection of pLR colonies, which remained readily available for subsequent isolation. Significance and Impact of the Study Our findings established an effective method for selective enrichment and detection of pLR strains. [ABSTRACT FROM AUTHOR]

Lactobacillus rhamnosus is a diverse Gram-positive species with strains isolated from different ecological niches. Here, we report the genome sequence analysis of 40 diverse strains of L. rhamnosus and their genomic comparison, with a focus on the variable genome. Genomic comparison of 40 L. rhamnosus strains discriminated the conserved genes (core genome) and regions of plasticity involving frequent rearrangements and horizontal transfer (variome). The L. rhamnosus core genome encompasses 2,164 genes, out of 4,711 genes in total (the pan-genome). The accessory genome is dominated by genes encoding carbohydrate transport and metabolism, extracellular polysaccharides (EPS) biosynthesis, bacteriocin production, pili production, the cas system, and the associated clustered regularly interspaced short palindromic repeat (CRISPR) loci, andmore than 100 transporter functions andmobile genetic elements like phages, plasmid genes, and transposons. A clade distribution based on amino acid differences between core (shared) proteins matched with the clade distribution obtained from the presence-absence of variable genes. The phylogenetic and variome tree overlap indicated that frequent events of gene acquisition and loss dominated the evolutionary segregation of the strains within this species, which is paralleled by evolutionary diversification of core gene functions. The CRISPR-Cas system could have contributed to this evolutionary segregation. Lactobacillus rhamnosus strains contain the genetic andmetabolic machinery with strain-specific gene functions required to adapt to a large range of environments.Aremarkable congruency of the evolutionary relatedness of the strains' core and variome functions, possibly favoring interspecies genetic exchanges, underlines the importance of gene-acquisition and loss within the L. rhamnosus strain diversification. [ABSTRACT FROM AUTHOR]

The human gastrointestinal epithelium makes up the largest barrier separating the body from the external environment. Whereas invasive pathogens cause epithelial barrier disruption, probiotic micro-organisms modulate tight junction regulation and improve epithelial barrier function. In addition, probiotic strains may be able to reduce epithelial barrier disruption caused by pathogenic species. The aim of this study was to explore non- Saccharomyces yeast modulation of epithelial cell barrier function in vitro. Benchmarking against established probiotic strains, we evaluated the ability of four nonpathogenic yeast species to modulate transepithelial electrical resistance ( TER) across a monolayer of differentiated human colonocytes (Caco-2 cells). Further, we assessed yeast modulation of a Salmonella Typhimurium-induced epithelial cell barrier function insult. Our findings demonstrate distinct patterns of non- Saccharomyces yeast modulation of epithelial cell barrier function. While the established probiotic yeast Saccharomyces boulardii increased TER across a Caco-2 monolayer by 30%, Kluyveromyces marxianus exhibited significantly stronger properties of TER enhancement (50% TER increase). In addition, our data demonstrate significant yeast-mediated modulation of Salmonella-induced epithelial cell barrier disruption and identify K. marxianus and Metschnikowia gruessii as two non- Saccharomyces yeasts capable of protecting human epithelial cells from pathogen invasion. Significance and Impact of the Study This study demonstrates distinct patterns of non- Saccharomyces yeast modulation of epithelial cell barrier function in vitro. Further, our data demonstrate significant yeast-mediated modulation of Salmonella Typhimurium-induced epithelial cell barrier disruption and identify Kluyveromyces marxianus and Metschnikowia gruessii as two non- Saccharomyces yeasts capable of protecting human epithelial cells from pathogen invasion. This study is the first to demonstrate significant non- Saccharomyces yeast-mediated epithelial cell barrier protection from Salmonella invasion, thus encouraging future efforts aimed at confirming the observed effects in vivo and driving further strain development towards novel yeast probiotics. [ABSTRACT FROM AUTHOR]

A specific probiotic formulation composed of Lactobacillus acidophilus CL1285, Lactobacillus casei LBC80R, and Lactobacillus rhamnosus CLR2 (Bio-K+) has been marketed in North America since 1996. The strains and the commercial products have been evaluated for safety, identity, gastrointestinal survival, and stability throughout shelf life. The capacity of both the fermented beverages and the capsules to reduce incidences of antibiotic-associated diarrhea and Clostridium difficile infection (CDI) has been demonstrated in human clinical trials. Individual strains and the finished products have shown antimicrobial activity against C. difficile and toxin A/B neutralization capacity in vitro. The use of this specific probiotic formulation as part of a bundle of preventive measures to control CDI in healthcare settings is discussed. [ABSTRACT FROM AUTHOR]

In August 2003, the 284-bed community hospital Pierre-Le Gardeur (PLGH) in Quebec experienced a major outbreak associated with the Clostridium difficile NAP1/027/BI strain. Augmented standard preventive measures (SPMs) were not able to control this outbreak. It was decided in February 2004 to give to every adult inpatient on antibiotics, without any exclusion, a probiotic (Bio-K+: Lactobacillus acidophilus CL1285, Lactobacillus casei LBC80R, and Lactobacillus rhamnosus CLR2) within 12 hours of the antibiotic prescription. Augmented SPMs were continued. The use of the probiotic in addition to SPMs was associated with a marked reduction of C. difficile infection (CDI). During the 10 years of observation, 44 835 inpatients received Bio- K+, and the CDI rate at PLGH declined from 18.0 cases per 10 000 patient-days and remained at low mean levels of 2.3 cases per 10 000 patient-days. Additionally, 10-year data collected by the Ministry of Health in Quebec comparing the CDI rate between Quebec hospitals showed that CDI rates at PLGH were consistently and continuously lower compared with those at similar hospitals. Blood cultures were monitored at PLGH for Lactobacillus bacteremia through the 10 years' experience, and no Lactobacillus bacteremias were detected. Despite the limitation of an observational study, we concluded that the probiotic Bio-K+ was safe and effective in decreasing our primary CDI rate. [ABSTRACT FROM AUTHOR]