Lactis BB-12 Research Articles

Pathogenic effects of Desulfovibrio in the gut on fatty liver in diet-induced obese mice and children with obesity.

Although we know the key role of gut dysbiosis in nonalcoholic fatty liver disease (NAFLD), it remains unclear what microbe(s) are responsible. This study aims to identify the microbes that cause NAFLD. C57BL/6JNarl male mice fed a high-fat diet (HFD) were orally administered Lactobacillus reuteri (L. reuteri) or Lactobacillus rhamnosus GG plus Bifidobacterium animalis subsp. lactis BB12 (LGG plus BB12). Their fecal microbiomes identified by 16S rRNA sequencing were correlated with the severity of fatty liver. We then used a human cohort to confirm the role of the microbe(s). The HFD-fed mice were administrated with the identified bacterium, Desulfovibrio. The histopathological changes in the liver and ileum were analyzed. Lactobacillus and Bifidobacterium improved hepatic steatosis and fibrosis in HFD-fed mice, which was related to the decreased abundance of Desulfovibrio in feces. Further human study confirmed the amount of D. piger in the fecal microbiota of obese children with NAFLD was increased. We then administered D. piger and found aggravated hepatic steatosis and fibrosis in HFD-fed mice. Hepatic expression of CD36 was significantly increased in HFD-fed mice gavaged with D. piger. In HepG2 cells, overexpression of CD36 increased lipid droplets, whereas knockdown of CD36 decreased lipid droplets. HFD-fed mice gavaged with D. piger had a decrease in the villus length, crypt depth, and zonula occludens-1 density in the ileum tissue. Our findings provide novel insights into the role of Desulfovibrio dysregulation in NAFLD. Modulation of Desulfovibrio may be a potential target for the treatment of NAFLD.

A fibre and phenolic-rich flour from Isabel grape by-products with stimulatory effects on distinct probiotics and beneficial impacts on human colonic microbiota invitro.

This study evaluated the effects of a fibre and phenolic-rich flour (IGF) prepared from Isabel grape by-products on the growth and metabolism of different probiotics and distinct bacterial populations part of the human intestinal microbiota during an invitro colonic fermentation. IGF was submitted to simulated gastrointestinal digestion before use in the experiments. IGF favoured the growth of the probiotics Lactobacillus acidophilus La-05, L. casei L-26 and Bifidobacterium lactis Bb-12, with viable counts of >7 log CFU per ml, as well as caused decreases in pH values and increases in organic acid production in the growth medium during 48 h of cultivation. IGF increased the population of beneficial micro-organisms forming the human intestinal microbiota, particularly Lactobacillus spp., decreased the pH values, and increased the lactic acid and short-chain fatty acid (acetic, butyric and propionic acids) production during 24 h of invitro colonic fermentation. These results indicate the potential prebiotic effects of IGF, which should represent a novel sustainable added-value ingredient with functional properties and gut-health benefits.

Effects of prebiotics on acid and bile resistance of Bifidobacterium lactis and Lactobacillus acidophilus probiotic bacteria

The effects of prebiotics on acid and bile resistance of probiotics were evaluated. Two commercial strains, Bifidobacterium lactis BB-12 and Lactobacillus acidophilus LA-5 had higher acid and bile resistance characteristics than the ATCC strains. B. lactis BB-12 was the most resistant to acid and bile and showed higher survival capacity. The counts of B. lactis BB-12 viable cells in pH 2 medium decreased by 2.7 log unit while the viability of other strains decreased by 4.5-7 log units. The counts of B. lactis BB-12 viable cells in the bile medium after 6 and 24 hours of incubation were determined as 6.1 and 2.8 log CFU/ml, respectively, while no viable bacteria could be determined for the other strains after 24 hours of incubation. The acid and bile resistance of probiotic bacteria remarkably varied with the type of prebiotics. Acid and bile resistance properties of L. acidophilus LA-5 and L. acidophilus ATCC 4356 were found higher when INU was present in the growth medium. Acid and bile resistance properties of B. bifidum ATCC 15969 was higher when the medium contained GOS whereas acid and bile resistance of B. lactis BB-12 was found to be higher when medium contained XOS.

Optimization and characterization of prebiotic concentration of edible films containing Bifidobacterium animalis subsp. lactis BB-12® and its application to block type processed cheese

The optimum inulin (IN) and fructooligosaccharide (FOS) concentration in sodium caseinate-based edible films containing Bifidobacterium animalis subsp. lactis BB-12® was studied and their use for the coating of sliced processed cheese was investigated. Physicochemical, optical, and mechanical properties of films were significantly affected by IN and FOS concentrations. Furthermore, the prebiotics generally increased the drying stability of the probiotic. The best film formulations were determined as 0% IN + 0% FOS, 1% IN + 0% FOS, and 0% IN + 1% FOS, respectively. Coating applications were effective in reducing weight loss and preserving the softness and colour properties of cheese. Probiotic viability was not affected by the IN and FOS during the drying of the coating on the cheese slice. It was determined that the counts of probiotics in the coated cheeses decreased over time and the concentration of viable probiotics in coated cheeses was insufficient after 30 days.

A Four-Probiotic Regime to Reduce Surgical Site Infections in Multi-Trauma Patients.

Investigations that focused on the protective role of probiotics against Surgical Site Infections (SSI) in multiple-trauma (MT) patients are generally few, probably due to the complexity of the concept of trauma. We aimed to assess the efficacy of a four-probiotic regime to reduce the incidence of SSI in MT patients, with a brain injury included. MT patients, being intubated and expected to require mechanical ventilation for >10 days, were randomly allocated into placebo (n = 50) or probiotic treatment (n = 53) comprising Lactobacillus acidophilus LA-5 (1.75 × 109 cfu), Lactiplantibacillus plantarum UBLP-40 (0.5 × 109 cfu), Bifidobacterium animalis subsp. lactis BB-12 (1.75 × 109 cfu), and Saccharomyces boulardii Unique-28 (1.5 × 109 cfu) in sachets. All patients received two sachets of placebo or probiotics twice/day for 15 days and were followed-up for 30 days. The operations were classified as neurosurgical, thoracostomies, laparotomies, orthopedics, and others; then, the SSI and the isolated pathogen were registered. A total of 23 (46.0%) and 13 (24.5%) infectious insults in 89 (50 placebo patients) and 88 (53 probiotics-treated) operations (p = 0.022) were recorded, the majority of them relating to osteosynthesis—17 and 8, respectively. The most commonly identified pathogens were Staphylococcus aureus and Acinetobacter baumannii. Our results support published evidence that the prophylactic administration of probiotics in MT patients exerts a positive effect on the incidence of SSI.

A Novel Combination of Pre-, Pro-, and Postbiotics Demonstrates Protective Effects on Intestinal Permeability in an In Vitro Gut Barrier Model

ObjectivesTo test the effects of a novel product combining probiotics (Lactobacillus rhamnosus GG and Bifidobacterium animalis subsp. lactis BB-12), a prebiotic (bacteriophage blend), and a postbiotic (tributyrin), and its individual components, on intestinal permeability in an in vitro gut barrier model.MethodsShort term colonic incubation was performed for the individual components, the full product, and a blank control under conditions representative of the proximal large intestine, using the fecal microbiota of a single healthy human donor. Colonic batch suspensions were collected after 48 hours of incubation and filter sterilized. Caco-2 cell monolayers were placed on top of PMA-differentiated THP1-BlueTM cells, creating an in vitro model of intestinal barrier disruption. Colonic fermentation samples were diluted in complete medium and given apically to Caco-2/THP1-BlueTM co-cultures for 24 hours. Intestinal permeability was assessed with transepithelial electrical resistance (TEER). TEER was measured in Caco-2 monolayers at baseline and 24 hours after pre-treatment of Caco-2/THP-1 BlueTM co-cultures. Each final TEER value was normalized to its corresponding initial value.ResultsApplication of colonic suspensions from the combination of pre-, pro-, and postbiotics significantly increased TEER values compared to control (p < 0.05). Probiotics and tributyrin were also found to increase TEER (p < 0.05) but no significant effect was observed for prebiotic phages (p > 0.05).ConclusionsColonic batch fermentations of a novel combination of pre-, pro-, and postbiotics showed improved gut barrier function in vitro, indicated by increased TEER values. Results suggest that probiotic strains and tributyrin likely modulate these effects, consistent with previous research linking the probiotic LGG and short chain fatty acid butyrate to gut barrier protection.Funding SourcesThe study was conducted by an independent lab, ProDigest, and funded and sponsored by Ritual (Natals, Inc.). Compound Solutions, Inc. also financially supported the study.

Synergistic Effect of a Novel Combination of Pre-, Pro-, and Postbiotics on Microbial Fermentation and Metabolite Production In Vitro

Abstract Objectives To test the effect of probiotic strains (Lactobacillus rhamnosus GG and Bifidobacterium animalis subsp. lactis BB-12; PS), a prebiotic (bacteriophage blend; BB), a postbiotic (tributyrin; TB), or a combination of all three (full mix; FM) on microbial fermentation and metabolite production in vitro. Methods Short term colonic incubation was performed under conditions representative of the proximal large intestine, using the fecal microbiota of a single healthy human donor. At the start of the experiment, treatments and a blank control (BC), were added to the reactors. Each reactor also received a carbohydrate-supplemented nutritional medium and freshly prepared fecal inoculum. Reactors were closed with a rubber septum, made anaerobic through flushing with nitrogen, and incubated for 48 hours. Each condition was run in triplicate. Overall fermentative activity (pH and gas production), changes in microbial metabolite production (short chain fatty acid, lactate, and ammonium analysis) were assessed. Results In the first 6 hours, microbial fermentation occurred in all treatments, indicated by pH shifts and production of gas and lactate. FM, PS, and BB stimulated production of lactate compared to BC (p &amp;lt; 0.05). After 24 hours, FM, PS and TB stimulated production of butyrate compared to BC (p &amp;lt; 0.05). FM and PS yielded more lactate than either BB or TB (p &amp;lt; 0.05). After 48 hours, FM, PS, BB, and TB each stimulated production of butyrate compared to BC (p &amp;lt; 0.05). Additionally, FM yielded significantly more butyrate than PS, TB, or BB (p &amp;lt; 0.05). Conclusions All treatments resulted in similar pH profiles and were representative of ideal conditions for bacterial growth and metabolism. The increased production of lactate in FM and PS was attributed to the fermentative activities of probiotics LGG and BB-12. Each treatment stimulated production of butyrate compared to BC but the largest increase in butyrate production was seen in FM, indicating a synergistic relationship between pre-, pro-, and postbiotics. Funding Sources The study was conducted by an independent lab, ProDigest, and funded and sponsored by Ritual (Natals, Inc.). Compound Solutions, Inc. also financially supported the study.

A Novel Combination of Pre-, Pro-, and Postbiotics Modulates Bacterial Diversity and Composition In Vitro

ObjectivesTo test the effect of probiotic strains (Lactobacillus rhamnosus GG and Bifidobacterium animalis subsp. lactis BB-12), a prebiotic (bacteriophage blend), and a postbiotic (tributyrin) on the human intestinal microbiota in vitro.MethodsShort term colonic incubation was performed under conditions representative of the proximal large intestine, using the fecal microbiota of a single healthy human donor. At the start of the experiment, treatments and a blank control were added to the reactors. Each reactor also received a carbohydrate-supplemented nutritional medium and freshly prepared fecal inoculum. Reactors were closed with a rubber septum, made anaerobic through flushing with nitrogen, and incubated for 48 hours. Each condition was run in triplicate. Changes in microbial community composition were determined via 16S-targeted Illumina sequencing and flow cytometry. Enriched bacterial genera were identified using LEfSe (LDA > 2, p < 0.05).ResultsA significant increase in alpha-diversity was observed by 48h compared to the control. LEfSe revealed that community shifts were attributed to a wide range of bacterial genera including Gemmiger, Lactobacillus, Bacteroides, Eubacterium, Oscillibacter, and Holdemanella at 24h and of genera such as Lactobacillus, Bacteroides, Muribaculum, and Holdemanella at 48h. LGG and BB-12 were not detected in the donor microbiota originally but abundance of both was higher after incubation.ConclusionsThe results from this study suggest that a combination of pre-, pro-, and postbiotics modulates gut microbiota by increasing microbial diversity and the relative abundance of a number of bacterial genera. An increase in the abundance of LGG and BB-12 indicates growth of these strains in the presence of a human gut microbiota. Future human clinical studies are needed to further investigate the observed effects.Funding SourcesThe study was conducted by an independent lab, ProDigest, and funded and sponsored by Ritual (Natals, Inc.). Compound Solutions, Inc. also financially supported the study.

Survival and Release Behavior of a Novel Pre-, Pro-, and Postbiotic Capsule in an In Vitro SHIME Model

ObjectivesTo test the survival and release of probiotics (Lactobacillus rhamnosus GG and Bifidobacterium animalis subsp. lactis BB-12), a prebiotic (bacteriophage blend), and a postbiotic (tributyrin) in a capsule-within-capsule design during passage through the gastrointestinal tract under fed and fasted conditions.MethodsUsing an adapted Simulator of the Human Intestinal Microbial Ecosystem (SHIME®) model, physiological conditions of the stomach and small intestine, including pH profiles, temperature, incubation time, and addition of bile liquid and digestive enzymes, were simulated within the same reactor over time. The capsule contents included an inner capsule, containing probiotics and prebiotic phages, and an outer capsule, containing the postbiotic, with different delivery and release characteristics. Probiotic survival was assessed via plate counts and PMA-qPCR, release of phages was assessed via phage titer assay, and tributyrin release was assessed via gas chromatography. All experiments were performed in triplicate to account for biological variability.ResultsUnder fasted conditions, the majority of the probiotics bypassed the stomach and small intestine in a viable form, of which more than 11 billion CFU were culturable. There were no significant differences between fasted and fed states. Similarly, the majority of the prebiotic phages survived transit through the stomach and small intestine. Tributyrin was mostly released in the small intestine.ConclusionsThe capsule design ensured probiotics and prebiotic phages survived the harsh conditions of the upper gastrointestinal tract for delivery to the colon. The release of tributyrin within the small intestine supports its breakdown by pancreatic enzymes to produce short chain fatty acid butyrate. The findings of this study suggest a novel, colon-targeted probiotic delivery system explained by specific capsule and content characteristics.Funding SourcesThe study was conducted by an independent lab, ProDigest, and funded and sponsored by Ritual (Natals, Inc.). Compound Solutions, Inc. also financially supported the study.

“Nourish to Flourish”: complementary feeding for a healthy infant gut microbiome—a non-randomised pilot feasibility study

BackgroundThe introduction of complementary foods and changes in milk feeding result in modifications to gastrointestinal function. The interplay between indigestible carbohydrates, host physiology, and microbiome, and immune system development are areas of intense research relevant to early and later-life health.MethodsThis 6-month prospective non-randomised feasibility study was conducted in Auckland, New Zealand (NZ), in January 2018. Forty parents/caregivers and their infants were enrolled, with 30 infants allocated to receive a prebiotic NZ kūmara (flesh and skin; a type of sweet potato) prepared as a freeze-dried powder, and ten infants allocated to receive a commercially available probiotic control known to show relevant immune benefits (109 CFU Bifidobacterium lactis BB-12®). The primary outcome was the study feasibility measures which are reported here.ResultsRecruitment, participant retention, and data collection met feasibility targets. Some limitations to biological sample collection were encountered, with difficulties in obtaining sufficient plasma sample volumes for the proposed immune parameter analyses. Acceptability of the kūmara powder was met with no reported adverse events.ConclusionThis study indicates that recruiting infants before introducing complementary foods is feasible, with acceptable adherence to the food-based intervention. These results will inform the protocol of a full-scale randomised controlled trial (RCT) with adjustments to the collection of biological samples to examine the effect of a prebiotic food on the prevalence of respiratory tract infections during infancy.Trial registrationAustralia New Zealand Clinical Trials Registry ACTRN12618000157279. Prospectively registered on 02/01/2018.

Barley β-glucan for conjugated linoleic acid (CLA) production by Bifidobacterium animalis subsp. Lactis: Fatty acid variation and bacterial viability study

In the present study, production of two isomers (cis-9, trans-11 and trans-10, cis-12) of conjugated linoleic acid (CLA) by Bifidobacterium animalis subsp. Lactis BB12 was investigated in organic and conventional milk in the presence of barley derived β-glucan. Fatty acid profile, acidification profile, cell enumeration and lactose content were determined for fermented conventional and organic milk, before and after 30 h fermentation in 30 °C. Viscometery and sensory evaluations were also performed. The results showed higher counts of Bifidobacterium animalis subsp. Lactis BB12, and viscosity in organic milk. Also, organic fermented milk offered higher CLA as well as polyunsaturated fatty acids values and a lower level of saturated fatty acids in comparison to conventional fermented milk. Cis-9, trans-11 CLA, trans-10, cis-12 CLA and polyunsaturated fatty acids values, bacterial count and viscosity in organic milk containing β-glucan were 28%, 39%, 33%, one log CFU/mL and 59% higher than the samples without β-glucan, respectively. Although these results were obtained for conventional milk, the variation percentage in the organic milk was significantly more than the conventional milk. In addition, the total lactose content in organic and conventional milk containing β-glucan was 19% and 12% lower than the samples without prebiotic.

Changes in the Gut Metabolic Profile of Gestational Diabetes Mellitus Rats Following Probiotic Supplementation.

The roles of gut microbiota and metabolomics in women with gestational diabetes mellitus (GDM) are not well understood. This study investigated the gut metabolomic profiling of GDM rats and GDM rats treated with probiotic supplements. Associations between gut metabolites and microbiota were also studied in GDM rats. Liquid chromatography–mass spectrometry was used to detect gut metabolites in GDM rats and GDM rats treated with probiotic supplements of 0.5 g (low-dose group) or 1 g (high-dose group) for 15 days. Each gram of probiotic supplement contained 5 × 107 colony-forming units (CFU) of Lactobacillus rhamnosus LGG and 1 × 108 CFU of Bifidobacterium animalis subspecies lactis Bb12. The association between gut metabolites and microbiota in GDM rats was investigated using Spearman’s correlation. Finally, 10 rats in the normal pregnant group, eight rats in the GDM model group, eight GDM rats in the low-dose probiotics group, and nine GDM rats in the high-dose probiotics group were further studied. Serum parameters and pancreatic and colon histology were significantly changed in GDM rats, and these were restored using probiotic supplements. In total, 999 gut metabolites were detected in the feces, and GDM rats were distinguished from normal rats. The levels of 44 metabolites were increased in GDM rats, and they were alleviated using probiotic supplements. Changes in metabolites in GDM rats were associated with amino acids and bile acids metabolism signaling pathways. Furthermore, changes in metabolites after probiotic supplementation were associated with porphyrin and chlorophyll metabolism pathways. We found that the Allobaculum genus displayed strong positive correlations, whereas the Bryobacter and Gemmatimonas genera displayed strong negative correlations with metabolisms of amino acids and bile acids in GDM rats. The Lactobacillus and Bifidobacterium genera were positively correlated with gut metabolites. Overall, our results showed that metabolism signaling pathways of amino acids and bile acids are associated with the development of GDM. Probiotic supplements alleviate the pathology of GDM through the metabolism pathways of amino acids, bile acids, porphyrin, and chlorophyll.

Anti-Obesity Effects of Synbiotic Fermented Milk Supplement with Aloe Vera Gel

The aim of this study was to investigate the anti-obesity effects of Synbiotic fermented milk supplemented with aloe vera gel (AVG) in male Rattus norvegicus rats obese induced by a high fat diet. 28 rats were divided into 4 groups (C-) fed a standard diet SD. (C+) fed a high fat diet (HFD) with oral dosing 1.5 ml/day distilled water. (T1) were fed HFD with an oral dose of 1.5 ml/day yogurt supplemented with 5% AVG. (T2) were fed HFD with an oral dose of 1.5 ml/day Fermented milk with Bif. Lactis BB-12 supplemented with 5% AVG. The experiment lasted 8 weeks, during which the increase in body weight, food intake and food efficiency rate were calculated, and then the relative weight of internal organs, lipid profile and liver enzymes AST and ALT were estimated. The results indicated that there was a significant increase in the final body weight and the relative weight of the liver in the (C+) group compared to the (C-) group. A significant decrease in body weight was also observed in the two groups T1 and T2 compared to the (C+) group, despite being fed the same HFD, while there were no significant differences in food intake. The T2 group showed the highest anti-obesity ability as well as maintaining the lipid profile within the normal limits, which contributed to the reduction of TC, TG, LDL-C, VLDL-C which were 81.5±6.863, 84.83±9.283, 33.48±6.349 and 16.97±1.857 mg/dL respectively, and an increase in the level of HDL-C, which was 31.05 ±3.774, that led to a decrease in atherogenic index which was 1.656 ± 0.383. It also contributed to keeping the liver enzymes AST, ALT within the normal limit, which was 53.5 and 41.5 IU/L compared to the (C +) group that was fed the same HFD diet.

Dietary Postbiotics Reduce Cytotoxicity and Inflammation Induced by Crystalline Silica in an In Vitro RAW 264.7 Macrophage Model.

Crystalline silica (cSiO2) particles are naturally existing environmental toxicants. Exposure to cSiO2 could cause local or systemic inflammation and aggregate inflammation-associated diseases. Dietary postbiotics are reported to possess anti-inflammatory activities; however, their effects on cSiO2-triggered inflammation are unknown. Here, we investigate the impact of postbiotics from Lacticaseibacillus rhamnosus (LGG), Limosilactobacillus reuteri (L.reu), and Bifidobacterium animalis subsp. lactis Bb12 (BB12) on cSiO2-induced cytotoxicity and IL-1 cytokines in vitro using macrophages. The postbiotics used in this study were cell-free fractions of a probiotic growth medium collected at different time points. The in vitro model used was the wild-type murine macrophage RAW 264.7 cell line stably transfected with the inflammasome adapter protein, ASC. Our results indicate that all the postbiotics could reduce cSiO2-induced cytotoxicity in the wild-type and ASC macrophages and the effects were OD-dependent. Following priming with a lipopolysaccharide, cSiO2 treatment resulted in robust inflammasome activation in ASC, as reflected by the IL-1β release. These responses were minimal or absent in the wild-type RAW cells. All the postbiotics decreased the release of IL-1β from ASC; however, only LGG and BB12 reduced the IL-1β secretion from wild-type cells. Only the L.reu postbiotics reduced the IL-1α release from ASC. We conclude that the postbiotics from LGG, BB12, and L.reu can protect macrophages against cSiO2-induced cytotoxicity and suppress IL-1β activation.

Production of short chain fatty acids and vitamin B12 during the in-vitro digestion and fermentation of probiotic chocolate

The prebiotic functions of chocolate to improve the function and health benefit of the chocolate when enriched with probiotics has not been fully investigated. This study investigated the role of chocolates enriched with probiotics in the production of SCFAs and vitamin B12 during the in-vitro colonic fermentation. Seven probiotic bacteria were encapsulated using a mixture of cocoa powder (10%), FOS (2%) and Na-alginate (1%) before added to chocolates. The results revealed that encapsulated Lactobacillus plantarum and Bifidobacterium animalis spp. lactis BB12 produced significantly higher amounts (1876.5 ± 105.16 and 1348.51 ± 77.37 mmol, respectively) of acetic acid after 48h of colonic fermentation. Chocolates with encapsulated BB12, La5, Lactobacillus sanfranciscensis and Streptococcus thermophilus yielded higher amounts of propionic, isobutyric, butyric and isovaleric acid. Probiotic-chocolates with 70% cocoa produced significantly more acetic, propionic, and butyric acids than that of 45% cocoa. However, Probiotic-chocolates with 45% cocoa produced greater amounts of vitamin B12 than 70% cocoa. These data demonstrate that probiotic-chocolate could be a functional snack with additional health benefits.

Sensory and microbiological evaluation of probiotic yoghurt made with different types of probiotic cultures starter Lactobacillus acidophilus LA-5 ® and Bifidobacterium animalis subsp. lactis BB-12®

The incorporation of probiotic bacteria into yoghurt is believed could increase its health benefits such as improving intestinal health, reducing the risk of type 2 diabetes mellitus and more. The objective of this study was to evaluate the sensory and microbiological characteristics of probiotic yoghurt (PY) made with 2 types of starter cultures namely direct vat set (DVS) cultures and heirloom (HC) cultures. Lactobacillus acidophilus LA5 ® and Bifidobacterium animalis subsp. lactis (BB-12® ) used in this study were obtained from Chr. Hansen. Conventional yoghurt (CY) was made with Lactobacillus delbrueckii subsp. bulgaricus (Lb) and Streptococcus thermophilus (St). As much as 2% of starter cultures were added into 10% skimmed milk suspension with 10% of sucrose. Lb:St ratio in CY was 1:1, whereas Lb:St:LA-5:BB-12 ratio in PY was 1:1:2:2. The mixture was incubated at 38°C for 6-8 hrs. Total lactic acid bacteria and bifidobacteria, as well as pH value, were determined at 0, 1st, 2nd, 3rd, 4th week of storage. The sensory properties of yoghurt were evaluated at the 0th week and the 4th week. The results showed that after 4 weeks of storage at 4°C, the total lactic acid bacteria were slightly decreased ranging from 0.26 log CFU/mL (DVS-PY) to 0.79 log CFU/mL (CY), while total bifidobacteria was slightly decreased from 0.40 to 0.58 log CFU/mL. The pH value of CY was significantly decreased after 4 weeks of storage, while the pH values of DVS-PY and HC-PY were not significantly different. The result from the Triangle test revealed that DVS-PY has similar sensory properties with CY whereas HC-PY has less thickness. In general, the sensory properties of CY and PY were slightly decreased after 4 weeks of storage at 4°C. In conclusion, incorporation of probiotic bacteria in yoghurt with DVS starter culture give better microbiological and sensory properties compared to HC starter culture.

EFFECT OF TIME AND INCUBATION TEMPERATURE ON ABILITY OF PROBIOTICS FOR REMOVAL OF POLYCYCLIC AROMATIC HYDROCARBON IN PHOSPHATE BUFFER SALINE

The ability of Lactobacillus acidophilus LA-5, Bifidobacterium lactis BB-12, Lactobacillus delbrueckii subsp. bulgaricus PTCC 1737 and Streptococcus thermophilus PTCC 1738 in removing Benzo(a)pyrene (BaP), Benz(a)anthracene (BaA), Chrysene (Chr) and Benzo(b)fluoranthene (BbF) from contaminated aqueous solution with respect to strain types, time and temperature of incubation was studied. The results showed that BaA, Chr, BbF and BaP could be significantly removed from the phosphate buffer saline by the tested bacteria and this reduction significantly depends on strain types and incubation temperature and time. The reduction of four PAHs varied in the range from 50.94% to 74.51%, 54.48% to 77.98%, 47.35% to 68.84%, and 61.96% to 81.22% for BaA, Chr, BbF and BaP, respectively. The results illustrated that by increasing time and temperature of incubation, binding ratio increased and the highest binding ratio of four PAHs was achieved by L. acidophilus LA-5 at 37 °C. Furthermore, the FTIR analysis was performed for identifying the different functional groups that might participate in the binding of PAHs to bacterial cell wall. The FTIR revealed that cell wall has an important role in removal of BaA, Chr, BbF and BaP from phosphate buffer saline.

The Effect of Essential Oils on the Survival of Bifidobacterium in In Vitro Conditions and in Fermented Cream

Essential oils derived from plant materials are a mixture of compounds that exhibit antibacterial properties. Due to their distinct aroma, they also serve as a desirable natural additive for various food products, including dairy products. In this study, the essential oils of lemon peels, clove buds, and juniper berries were obtained by steam distillation and characterized using gas chromatography–mass spectrometry to determine their chemical compositions and effects on the viability of seven Bifidobacterium strains. Furthermore, the effect of essential oils on the viability of Bifidobacterium animalis subsp. lactis Bb-12 was investigated in cream samples during fermentation and after storage for 21 days at 6 °C. The fatty acid composition of fat extracted from essential oils containing sour cream samples and the volatile aroma compound profile of the sour cream samples were also determined chromatographically. Among the 120 compounds identified, monoterpene hydrocarbons were dominant in the essential oils of lemon peels (limonene and γ-terpinene) and juniper berries (sabinene and β-myrcene), while eugenol and eugenol acetate were abundant in the essential oil of clove buds. In addition to these compounds, butanoic and acetic acids were found in the tested sour cream samples. In turn, fat extracted from these samples was rich in saturated fatty acids, mainly palmitic acid. Among the tested strains of the genus Bifidobacterium, B. animalis subsp. lactis Bb-12 was the most sensitive to the essential oils of clove and juniper, as indicated by the larger growth inhibition zones. However, both the concentration and type of essential oils used had no effect on the number of cells of this strain present in the cream samples immediately after fermentation and after its 21-day storage, which suggests that the tested essential oils could be a natural additive to dairy products.

Anti-rotavirus Properties and Mechanisms of Selected Gram-Positive and Gram-Negative Probiotics on Polarized Human Colonic (HT-29) Cells.

Probiotics have been investigated to improve the universal rotavirus (RV) vaccination as well as to ameliorate the RV infection. However, underlying mechanisms how probiotics mediate beneficial effects needs more investigation. Thus, in the present study, we used polarized HT-29 cells to assess the anti-RV properties of Gram-positive, (Lactobacillus acidophilus, Lacticaseibacillus rhamnosus GG, and Bifidobacterium subsp. Lactis Bb12) and Gram negative, (Escherichia coli Nissle 1917) probiotics and study their underlying mechanisms. Our results showed that pre-treatment of HT-29 cells for 4h with probiotics, significantly reduced (p < 0.05) human RV replication and this effect was most pronounced for E. coli Nissle followed by L. acidophilus and L. rhamnosus GG. Strikingly, only pre-treatment with live bacteria or their supernatants demonstrated anti-RV properties. Except Gram negative E. coli Nissle, the Gram-positive probiotics tested did not bind to RV. Ingenuity pathway analysis of tight junction (TJ)- and innate immune-associated genes indicated that E. coli Nissle or E. coli Nissle + RV treatments improved cell-cell adhesion and cell contact, while L. acidophilus or L. acidophilus + RV treatments also activated cell-cell contact but inhibited cell movement functions. RV alone inhibited migration of cells event. Additionally, E. coli Nissle activated pathways such as the innate immune and inflammatory responses via production of TNF, while RV infection activated NK cells and inflammatory responses. In conclusion, E. coli Nissle's ability to bind RV, modulate expression of TJ events, innate immune and inflammatory responses, via specific upstream regulators may explain superior anti-RV properties of E. coli Nissle. Therefore, prophylactic use of E. coli Nissle might help to reduce the RV disease burden in infants in endemic areas.

Bifidobacterium animalis subsp. lactis BB-12 Has Effect Against Obesity by Regulating Gut Microbiota in Two Phases in Human Microbiota-Associated Rats.

Bifidobacterium animalis subsp. lactis BB-12 (BB-12) is an extensively studied probiotics species, which has been reported to improve the human gut microbiota. This study aimed to confirm the effects of BB-12 on high-fat diet (HFD)-induced gut microbiota disorders. The probiotic BB-12 was consumed by human microbiota-associated rats and changes in gut microbiota were compared using next generation sequencing of the fecal samples collected from the normal chow group, the HFD group, and the BB-12-supplemented group. The enterotypes switched from Prevotella dominant to Akkermansia dominant as a result of switching diet from normal chow to HFD. BB-12 conferred protection on the gut microbiota composition of the rats by increasing the abundance of Prevotella and decreasing the abundance of Clostridium, Blautia, and Bacteroides in 0–3 weeks. In addition, Prevotella-dominant enterotype was maintained, which provides improve obesity effects. A decrease in body weight and the Firmicutes/Bacteroidetes ratio were also observed at week 3. While in 4–8 weeks, the enrichment of short-chain fatty acids-producing bacteria such as Eubacterium and Parabacteroides and probiotics such as Bifidobacterium was observed. The results revealed that BB-12 against obesity by regulating gut microbiota in two phases. After a short-term intervention, BB-12 supplementation suppressed the transition from the healthy to obesity state by protecting Prevotella-dominant enterotype, whereas after a long-term intervention, BB-12 ameliorates obesity by enriching beneficial bacteria in the gut.