Probiotic interventions promote metabolic health in high fat-fed hamsters in association with gut microbiota and endocannabinoidome alterations.

Probiotics represent a promising tool to improve metabolic health, including lipid profiles and cholesterol levels. Modulation of the gut microbiome and the endocannabinoidome - two interrelated systems involved in several metabolic processes influenced by probiotics - has been proposed as a potential mechanism of action. This study establishes the impact of probiotics on metabolic health, gut microbiota composition and endocannabinoidome mediators in an animal model of hypercholesterolaemia. Syrian hamsters were fed either a low-fat low-cholesterol or high-fat high-cholesterol (HFHC) diet to induce hypercholesterolaemia and gavaged for 6 weeks with either Lactobacillus acidophilus CL1285, Lactiplantibacillus plantarum CHOL-200 or a combination of the two. Globally, probiotic interventions ameliorated, at least partially, lipid metabolism in HFHC-fed hamsters. The interventions, especially those including L. acidophilus, modified the gut microbiota composition of the small intestine and caecum in ways suggesting reversal of HFHC-induced dysbiosis. Several associations were observed between changes in gut microbiota composition and endocannabinoidome mediators following probiotic interventions and both systems were also associated with improved metabolic health parameters. For instance, potential connexions between the Eubacteriaceae and Deferribacteraceae families, levels of 2­palmitoylglycerol, 2­oleoylglycerol, 2­linoleoylglycerol or 2­eicosapentaenoylglycerol and improved lipid profiles were found. Altogether, our results suggest a potential crosstalk between gut microbiota and the endocannabinoidome in driving metabolic benefits associated with probiotics, especially those including L. acidophilus, in an animal model of hypercholesterolaemia.

Primary prevention of Clostridium difficile infections with a specific probiotic combining Lactobacillus acidophilus, L. casei, and L. rhamnosus strains: assessing the evidence.

Clostridium difficile infection (CDI) has become the leading healthcare-associated infection and cause of outbreaks around the world. Although various innovative treatments have been developed, preventive strategies using multi-faceted infection control programmes have not been successful in reducing CDI rates. The major risk factor for CDI is the disruption of the normally protective gastrointestinal microbiota, typically by antibiotic use. Supplementation with specific probiotics has been effective in preventing various negative outcomes, including antibiotic-associated diarrhoea and CDI. However, a consensus of which probiotic strains might prevent CDI has not been reached and meta-analyses report high degrees of heterogeneity when studies of different probiotic products are pooled together. We searched the literature for probiotics with sufficient evidence to assess clinical efficacy for the prevention of CDI and focused on one specific probiotic formulation comprised of three lactobacilli strains (Lactobacillus acidophilus CL1285, Lactobacillus casei LBC80R, Lactobacillus rhamnosus CLR2, Bio-K+) for its ability to prevent CDI in healthcare settings. A literature search on this probiotic formulation was conducted using electronic databases (PubMed, Google Scholar), abstracts from infectious disease and infection control meetings, and communications from the probiotic company. Supporting evidence was found for its mechanisms of action against CDI and that it has an excellent safety and tolerability profile. Evidence from randomized controlled trials and facility-level interventions that administer Bio-K+ show reduced incidence rates of CDI. This probiotic formulation may have a role in primary prevention of healthcare-associated CDI when administered to patients who receive antibiotics.

Probiotics and prevention of Clostridium difficile infection.

The role of probiotics as adjunctive measures in the prevention of Clostridium difficile infection (CDI) has been controversial. However, a growing body of evidence has suggested that they have a role in primary prevention of CDI. Elements of this controversy are reviewed and the proposed mechanisms of action, the value and cost effectiveness of probiotics are addressed with a focus on three agents, Saccharomyces boulardii, Lactobacillus rhamnosus GG and the combination of Lactobacillus acidophilus CL1285, Lactobacillus casei LBC80R, Lactobacillus rhamnosus CLR2 (Bio-K+).

Impact of elective hospital admissions on glycaemic control in adolescents with poorly controlled type 1 diabetes.

AIM: Different treatment strategies have been used to manage adolescents with poorly controlled type 1 diabetes. We investigated whether a brief elective hospital admission improves haemoglobin A(1c) (HbA(1c)) over 12 months. METHODS: We studied a retrospective cohort of adolescents with poorly controlled type 1 diabetes attending a tertiary care pediatric diabetes clinic in Montreal, Canada, between January 2005 and December 2010. Hospitalized adolescents (admitted group) were matched with controls (non-admitted group) for age and baseline HbA(1c). HbA(1c) values at baseline, 6 and 12 months were obtained from the clinic database. RESULTS: Thirty patients aged 11 to 17 years with a first elective admission for poor metabolic control were paired with 30 non-admitted patients. At baseline, HbA(1c) was 12.2±1.6% in admitted and 12.0±1.2% in non-admitted patients. There were no clinically important differences in potential confounders between groups. There was no improvement in the primary outcome as assessed by the change in HbA(1c) at 12 months in the admitted group (-1.3±2.3%) compared with the non-admitted group (-2.1±1.7%) (P=0.078). No improvement in intermediary measures of glycaemic control was observed (HbA(1c) at 6 months or change at 6 months). After 12 months, HbA(1c) values were higher in the admitted group (10.9±1.9%) versus the non-admitted group (9.9±1.4%) (P=0.016). CONCLUSION: Elective hospital admission for adolescents with poorly controlled type 1 diabetes does not seem to be an effective strategy to improve HbA(1c) over 12 months.

Intermediate chains of exopolysaccharides from Lactobacillus rhamnosus RW-9595M increase IL-10 production by macrophages.

AIMS: To evaluate the immunosuppressive properties of the exopolysaccharide (EPS) from high-EPS producer Lactobacillus rhamnosus RW-9595M on inflammatory cytokines produced by macrophages. METHODS AND RESULTS: The conditioned media (CM) were produced by macrophages treated with parental Lact. rhamnosus ATCC 9595 and its isogenic variant, the high-EPS producer Lact. rhamnosus RW-9595M, and the levels of TNF-alpha, IL-6, IL-10 and IL-12 were evaluated. Results revealed that CM from parental Lact. rhamnosus induced higher levels of TNF-alpha, IL-6 and IL-12 but inhibited IL-10 production, whereas its mucous variant induced low or no TNF-alpha and IL-6. Addition of purified EPS to macrophages treated with parental Lact. rhamnosus decreased the inflammatory cytokines and inhibited the metabolic activity of lymphocytes. The intermediate polysaccharide chains (16-30 units) produced by time-controlled hydrolysis of EPS increased the IL-10 produced by macrophages. CONCLUSIONS: Polysaccharide chains of EPS induced immunosuppression by the production of macrophagic anti-inflammatory IL-10. SIGNIFICANCE AND IMPACT OF THE STUDY: These results indicate that the EPS from Lact. rhamnosus RW-9595M may be useful as a new immunosuppressive product in dairy food.

Effect of selected antimicrobial compounds on the radiosensitization of Salmonella Typhi in ground beef.

AIMS: In this study, we extended our previous work to determine the efficiency of antimicrobial compounds in increase of relative radiosensitivity of Salmonella Typhi in medium fat ground beef (23% fat) by testing 41 different essential oils (EOs), oleoresins and food sauces. METHODS AND RESULTS: Ground beef samples inoculated with Salmonella Typhi (10(6) CFU g(-1)) were treated with each antimicrobial compound at a concentration of 0.5% (w/w). Then, the samples (25 g each) were packaged under air and irradiated in a (60)Co irradiator at doses from 0 to 1.75 kGy. Radiosensitivity was evaluated by calculating relative radiation sensitivity, defined as the ratio of radiation D(10) value in the absence/presence of antimicrobial compound. CONCLUSIONS: Depending on the compound tested, the addition of antimicrobial compound decreased the D(10) value of Salmonella Typhi, resulting in an increase of the radiation sensitivity up to more than four times. Among these antimicrobial compounds, Chinese cinnamon EO, clove EO and trans-cinnamaldehyde were most effective to increase the radiosensitivity of Salmonella Typhi in ground beef. SIGNIFICANCE AND IMPACT OF THE STUDY: These observations demonstrate that some active compounds can function as radiosensitizers of Salmonella Typhi.

Effect of gamma radiation on heat shock protein expression of four foodborne pathogens.

AIMS: The effects of gamma radiation on three heat shock proteins (Hsps) (GroEL, DnaK and GroES) synthesis in two Gram-negative (Escherichia coli and Salmonella serotype Typhimurium) and two Gram-positive (Staphylococcus aureus and Listeria monocytogenes) bacteria were investigated. METHODS AND RESULTS: The bacterial strains were treated with three radiation doses to induce cell damage, to obtain a viable but nonculturable state, and to cause cell death. Western blot analysis and quantification of Hsps in bacteria were performed immediately after irradiation treatment. In the four foodborne pathogens, GroEL was strongly induced by gamma rays in a dose-dependent manner, confirming the involvement of this protein in the cellular response to the stress generated by ionizing radiation. In addition, it was found that E. coli exposed to gamma radiation showed a significantly induction of DnaK and GroES proteins when compared with nonirradiated bacteria, whereas a GroES slight induction and a DnaK inhibition were observed in Salm. Typhimurium. CONCLUSIONS: The gamma rays influence the synthesis of Hsps in foodborne pathogen in a way that critically depends on the radiation dose. SIGNIFICANCE AND IMPACT OF THE STUDY: The study of stress response to several radiation doses was undertaken to elucidate how bacteria can survive in harsh conditions and cope with gamma radiation used to control foodborne pathogens and to characterize their adaptative response to this treatment.

Effect of selected plant essential oils or their constituents and modified atmosphere packaging on the radiosensitivity of Escherichia coli O157:H7 and Salmonella typhi in ground beef.

Twenty-six different essential oils were tested for their efficiency to increase the relative radiosensitivity of Escherichia coli and Salmonella Typhi in medium-fat ground beef (23% fat). Ground beef was inoculated with E. coli O157:H7 or Salmonella (10(6) CFU/g), and each essential oil or one of their main constituents was added separately at a concentration of 0.5% (wt/wt). Meat samples (10 g) were packed under air or under modified atmosphere and irradiated at doses from 0 to 1 kGy for the determination of the D10-value of E. coli O157:H7, and from 0 to 1.75 kGy for the determination of the D10-value of Salmonella Typhi. Depending on the compound tested, the relative radiation sensitivity increased from 1 to 3.57 for E. coli O157:H7 and from 1 to 3.26 for Salmonella Typhi. Addition of essential oils or their constituents before irradiation also reduced the irradiation dose needed to eliminate both pathogens. In the presence of Chinese cinnamon or Spanish oregano essential oils, the minimum doses required to eliminate the bacteria were reduced from 1.2 to 0.35 and from 1.4 to 0.5 for E. coli O157:H7 and Salmonella Typhi, respectively. Cinnamon, oregano, and mustard essential oils were the most effective radiosensitizers.

Serum dioxin levels in residents of Calcasieu and Lafayette parishes, Louisiana with comparison to the US population.

The Agency for Toxic Substances and Disease Registry (ATSDR) used a cross-sectional study to compare the serum dioxin toxic equivalent (TEQ) levels of a population-based representative sample of Calcasieu Parish residents aged 15 years and older to a similar group of residents of Lafayette Parish with less industrial facilities. Serum dioxins consisted of polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, and dioxin-like polychlorinated biphenyls. Overall, the mean and distribution of serum dioxin TEQ level in residents of both parishes were similar by age groups (15-29 years, 30-44 years, 45-59 years, and 60 year and older). When the Calcasieu Parish area was further divided based on distance to three industrial areas, the mean dioxin TEQ levels were similar. Serum dioxin TEQ levels in residents of both parishes increased with age. Calcasieu Parish residents who reported having eaten locally caught fish, smoked cigarettes, worked in an occupation with potential exposure, or used pesticides had dioxin levels similar to Lafayette Parish residents who reported these activities. African Americans had higher dioxin levels than Caucasians in Lafayette Parish and both races in Calcasieu Parish. The congener profiles were similar in residents of both parishes. When the combined Calcasieu and Lafayette Parish data were compared by age group to the National Health and Nutrition Examination Survey (NHANES) 2001-2002 data, the geometric means for the dioxin levels in the combined Parish data set were significantly lower than the NHANES data in all age groups (all P-values <0.0001), except the oldest age group where the significance level is marginal (P=0.067). The various percentiles of the youngest age group of the combined parish data were also significantly lower than those in NHANES. Since the combined parish dioxin levels were below a representative sampling of the US population, there is no increase in serum dioxin concentrations in both the parishes.

Purification and identification of the pediocin produced by Pediococcus acidilactici MM33, a new human intestinal strain.

AIMS: The aim of this study was to purify and identify the bacteriocin produced by Pediococcus acidilactici MM33, a strain previously isolated from human gut. METHODS AND RESULTS: Purification of the bacteriocin was performed by cationic exchange chromatography followed by a reverse phase step. Biochemical and mass spectrometry analysis showed homology with pediocin PA-1. To verify if P. acidilactici MM33 carried the pediocin PA-1 gene, total DNA was used to amplify the pediocin gene. The PCR product obtained was then sequenced and the nucleotide sequence revealed to be identical to that of pediocin PA-1. Treatment of P. acidilactici MM33 with novobiocin resulted in a plasmid-cured strain without bacteriocin-producing capacity. Antimicrobial assay and molecular analysis demonstrated that this strain was ped(-) suggesting that the ped cluster is plasmid encoded. Antimicrobial assay revealed that pediocin was bactericidal against Listeria monocytogenes, showing a minimal inhibitory concentration (MIC) of 200 AU ml(-1). CONCLUSIONS: A two-step purification procedure was elaborated in this study. The bacteriocin secreted by the human strain P. acidilactici MM33 is carried on a plasmid and the amino acid sequence is identical to pediocin PA-1. SIGNIFICANCE AND IMPACT OF THE STUDY: Pediococcus acidilactici MM33 is the first human pediocin-producing strain reported and could be used as probiotic to prevent enteric pathogen colonization.

In vitro growth control of selected pathogens by Lactobacillus acidophilus- and Lactobacillus casei-fermented milk.

AIMS: Food-borne pathogen inhibition was tested in the presence of a mixture of Lactobacillus acidophilus and Lactobacillus casei during fermentation under controlled pH conditions. METHODS AND RESULTS: The growth of Escherichia coli O157:H7, Salmonella serotype Typhimurium, Staphylococcus aureus, Listeria innocua, Enterococcus faecium and Enterococcus faecalis was evaluated for 48 h at 37 degrees C. In the presence of the lactic acid bacteria (LAB), an increase of the generation time was observed for all the gram-positive bacteria evaluated. Staphylococcus aureus was the most sensitive strain showing an increase of the generation time by 210%. However, for all the gram-negative bacteria evaluated, no inhibition occurred after 8 h of fermentation. The soluble portion of Lact. acidophilus- and Lact. casei-fermented milk was recuperated and tested for its antimicrobial activity. Listeria innocua and Staph. aureus were the most sensitive to the presence of fermented milk supernatant showing an inhibition of 85.9% and 84.7%, respectively. This soluble fraction was neutralized to eliminate the antimicrobial effect of the organic acids produced; the most sensitive strains were L. innocua and E. coli O157:H7 showing an inhibition of 65.9% and 61.9%, respectively. Finally, the soluble fraction was neutralized and irradiated at 45 kGy using a (60)Co source to eliminate the possible antimicrobial effect of both organic acids and bacteriocin-like substances. Enterococcus faecalis, E. coli O157:H7 and Staph. aureus were the most affected bacteria by this fraction, showing 39.1, 32 and 31.2% inhibition, respectively. CONCLUSIONS: The results obtained in this study suggest the implication of both organic acids and bacteriocin-like inhibitory substances in the antimicrobial activity observed in the soluble fraction of the probiotic preparation. SIGNIFICANCE AND IMPACT OF THE STUDY: This study revealed the antimicrobial mechanisms of action of Lact. acidophilus- and Lact. casei-fermented milk used to prevent antibiotic-associated diarrhoea.

Partial characterization of bacteriocins produced by human Lactococcus lactis and Pediococccus acidilactici isolates.

AIMS: The aim of this study was to isolate bacteriocin-producing lactic acid bacteria (LAB) from human intestine. METHODS AND RESULTS: A total of 111 LAB were isolated from human adult stool and screened for their bacteriocin production. Neutralized cell-free supernatants from Lactococcus lactis subsp. lactis MM19 and Pediococcus acidilactici MM33 showed antimicrobial activity. The antimicrobials in the supernatant from a culture of L. lactis inhibited Enterococcus faecium, various species of Lactobacillus and Staphylococcus aureus; while those in the supernatant from a culture of P. acidilactici inhibited Enterococcus spp., some lactobacilli and various serotypes of Listeria monocytogenes. The antimicrobial metabolites were heat-stable and were active over a pH range of 2-10. The antimicrobial activities of the supernatants of both bacteria were inhibited by many proteases but not by catalase. The plate overlay assay allowed an approximation of size between 3.5 and 6 kDa for both antimicrobial substances. CONCLUSIONS: As the antagonistic factor(s) produced by L. lactis MM19 and P. acidilactici MM33 were sensitive to proteolytic enzymes, it could be hypothesized that bacteriocins were involved in the inhibitory activities. Inhibition spectrum and biochemical analysis showed that these bacteria produced two distinct bacteriocins. SIGNIFICANCE AND IMPACT OF THE STUDY: We are the first to isolate bacteriocin-producing strains of Pediococcus and Lactococcus from human intestine. These strains might be useful for control of enteric pathogens.

Combined effects of antimicrobial coating, modified atmosphere packaging, and gamma irradiation on Listeria innocua present in ready-to-use carrots (Daucus carota).

The objective of this study was to evaluate the effect of an edible antimicrobial coating combined with modified atmosphere (MA) packaging (60% O2, 30% CO2, and 10% N2) and gamma irradiation on peeled minicarrots inoculated with Listeria innocua. Carrots were inoculated with L. innocua (10(3) CFU/g) and then coated with an antimicrobial coating based on calcium caseinate containing trans-cinnamaldehyde. The same formulation without trans-cinnamaldehyde was used as an inactive coating. Coated and uncoated carrots were packed under the MA or under air, irradiated at 0.25 or 0.5 kGy, and stored at 4 +/- 1 degrees C for 21 days. Samples were evaluated periodically for enumeration of L. innocua. Unirradiated carrots stored under air had the highest concentrations of L. innocua after 21 days of storage: 2.23 CFU/g in the uncoated samples and 2.26 CFU/ g in samples coated with the inactive coating. These results suggest that the inactive coating did not have any antimicrobial effect against L. innocua. However, the addition of the antimicrobial coating resulted in a 1.29-log reduction in the concentration of L. innocua in carrots packed under air after 21 days of storage and a 1.08-log reduction in carrots packed under MA after 7 days of storage. After 7 days of storage, no L. innocua was detected in samples treated at 0.5 kGy under air or in samples treated at 0.25 kGy under MA. A complete inhibition of L. innocua was also observed during all storage periods in uncoated and coated samples treated at 0.5 kGy under MA. These results indicate that the combination of irradiation and MA conditions play an important role in the radiosensitization of L. innocua.

Influence of antimicrobial compounds and modified atmosphere packaging on radiation sensitivity of Listeria monocytogenes present in ready-to-use carrots (Daucus carota).

Radiosensitization of Listeria monocytogenes was determined in the presence of trans-cinnamaldehyde, Spanish oregano, winter savory, and Chinese cinnamon on peeled minicarrots packed under air or under a modified atmosphere (60% O2, 30% CO2, and 10% N2). Samples were inoculated with L. monocytogenes HPB 2812 serovar 1/2a (106 CFU/g) and were coated separately with each active compound (0.5%, wt/wt) before being packaged under air or the modified atmosphere and irradiated at doses from 0.07 to 2.4 kGy. Results indicated that the bacterium was more resistant to irradiation under air in the absence of active compound. The dose required to reduce L. monocytogenes population by 1 log CFU (D10) was 0.36 kGy for samples packed under air and 0.17 kGy for those packed under the modified atmosphere. The active compounds evaluated in this study had an effect on the radiation sensitivity of L. monocytogenes on carrots. The most efficient compound was trans-cinnamaldehyde, where a mean 3.8-fold increase in relative radiation sensitivity was observed for both atmospheres compared with the control. The addition of winter savory and Chinese cinnamon produced a similar increase in relative radiation sensitivity but only when samples where packed under modified atmosphere conditions.

Communicating results to community residents: lessons from recent ATSDR health investigations.

As a public health agency within the US Department of Health and Human Services, the Agency for Toxic Substances and Disease Registry (ATSDR) is responsible for implementing the health-related provisions of the Superfund Act. Much of its work is carried out to address health concerns in communities near sources of environmental contamination, usually in consultation with other local, state, and federal agencies. Over the last decade, ATSDR has considered, supported or conducted health investigations in a variety of different communities across the country. Communication with community residents has been an integral part of the process in all of these activities. The approach to communicating results needs to begin early by developing relationships and clarifying expectations, and it needs to remain flexible. Through examples taken from specific situations, we illustrate many of the lessons we have gained from trying to apply the principles of good community involvement to the design and conduct of health investigations and to the communication of study results.

Antimicrobial potential of immobilized Lactococcus lactis subsp. lactis ATCC 11454 against selected bacteria.

Immobilization of living cells of lactic acid bacteria could be an alternative or complementary method of immobilizing organic acids and bacteriocins and inhibit undesirable bacteria in foods. This study evaluated the inhibition potential of immobilized Lactococcus lactis subsp. lactis ATCC 11454 on selected bacteria by a modified method of the agar spot test. L. lactis was immobilized in calcium alginate (1 to 2%)-whey protein concentrate (0 and 1%) beads. The antimicrobial potential of immobilized L. lactis was evaluated in microbiological media against pathogenic bacteria (Escherichia coli, Salmonella, and Staphylococcus aureus) or Pseudomonas putida, a natural meat contaminant, and against seven gram-positive bacteria used as indicator strains. Results obtained in this study indicated that immobilized L. lactis inhibited the growth of S. aureus, Enterococcus faecalis, Enterococcus faecium, Lactobacillus curvatus, Lactobacillus sakei, Kocuria varians, and Pediococcus acidilactici. Only 4 h of incubation at 35 degrees C resulted in a clear inhibition zone around the beads that increased with time. With the addition of 10 mM of a chelating agent (EDTA) to the media, results showed growth inhibition of E. coli; however, P. putida and Salmonella Typhi were unaffected by this treatment. These results indicate that immobilized lactic acid bacteria strains can be successfully used to produce nisin and inhibit bacterial growth in semisolid synthetic media.

The isotopic cardiac pacer: a ten-year experience.

In the past, the battery has been the main cause of pacemaker failure. The isotopic-powered cardiac pacemaker theoretically should provide greater reliability and longevity than pacemakers powered by chemical batteries. We studied 79 isotopic pacemakers in 75 patients for up to 10 years. Four types of Plutonium 238 powered pacemakers were studied. They included 5 VOO, 51 VVI, 14 VVIP, and 9 VVIM pacemakers. A more diverse group of 93 VVI, VVIP and VVIM pacemakers, powered by chemical batteries, were studied as controls. During the course of study the mercury zinc battery has become obsolete. The increased longevity of the lithium battery, at first very promising, has been compromised by the demands firstly of smaller size, and later of more complex circuits. The longevity and reliability of the isotopic pacemakers has been found to be superior to pacemakers powered by chemical batteries. The isotopic fuel cell, because of its greater longevity, may be needed for the increasingly complex circuits required for dual chamber pacemakers, functioning for the control of bradyarrhythmia alone, or combined with tachyarrhythmia control.