Genome-wide mining of gpx gene family provides new insights into cadmium stress responses in common carp (Cyprinus carpio).

Glutathione peroxidase (Gpx) is an important member of antioxidant enzymes, which can play a vital role in metabolizing reactive oxygen species (ROS) and in maintaining cell homeostasis. In order to study the evolutionary dynamics of gpx gene family in allotetraploid fish species, we identified a total of 14 gpx genes in common carp Cyprinus carpio, while 9 gpx genes were discovered in the diploid progenitor-like species Poropuntius huangchuchieni. Comparative genomic analysis and phylogenetic analysis revealed that the common carp gpx genes had significant expansion and were divided into five distinct subclades. Exon-intron distribution patterns and conserved motif analysis revealed highly conserved evolutionary patterns. Transcript profiles suggested that different gpx genes had specific patterns of regulation during early embryonic development. In adult tissues, gpx genes had a relatively broad expression distribution, most of which were highly expressed in the gills, intestines, and gonads. RT-qPCR studies showed that most gpx genes were downregulated during the initial cd2+ treatment stage. Dietary supplementation of Bacillus coagulans at different concentrations (Group 2 of 1.0 × 107 cfu/g, Group 3 of 1.0 × 108 cfu/g, and Group 4 of 1.0 × 109 cfu/g) induced different regulatory responses of gpx subclades. This result suggested that the appropriate concentration of B. coagulans can improve gpx gene expression when exposed to heavy metal cadmium treatment, which may play a vital role in the resistance to oxidative stress and immune responses. This study has expanded our understanding of the functional evolution of the gpx gene family in common carp.

The metabolism and morphology mutation response of probiotic Bacillus coagulans for lead stress.

Lead is among the most common toxic heavy metals and its contamination is of great public concern. Bacillus coagulans is the probiotic which can be considered as the lead absorption sorbent to apply in the lead contaminant water directly or indirectly. A better understanding of the lead resistance and tolerance mechanisms of B. coagulans would help further its development and utilization. Wild-type Bacillus coagulans strain R11 isolated from a lead mine, was acclimated to lead-containing culture media over 85 passages, producing two lead-adapted strains, and the two strains shown higher lead intracellular accumulation ability (38.56-fold and 19.36-fold) and reducing ability (6.94-fold and 7.44-fold) than that of wild type. Whole genome sequencing, genome resequencing, and comparative transcriptomics identified lead resistance and tolerance process significantly involved in these genes which regulated glutathione and sulfur metabolism, flagellar formation and metal ion transport pathways in the lead-adapted strains, elucidating the relationships among the mechanisms regulating lead deposition, deoxidation, and motility and the evolved tolerance to lead. In addition, the B. coagulans mutants tended to form flagellar and chemotaxis systems to avoid lead ions rather than export it, suggesting a new resistance strategy. Based on the present results, the optimum lead concentration in environment should be considered when employed B. coagulans as the lead sorbent, due to the bacteria growth ability decreased in high lead concentration and physiology morphology changed could reduce the lead removal effectiveness. The identified deoxidization and compound secretion genes and pathways in B. coagulans R11 also are potential genetic engineering candidates for synthesizing glutathione, cysteine, methionine, and selenocompounds.

Enriching novel dark chocolate with Bacillus coagulans as a way to provide beneficial nutrients.

The aim of the present study was to develop a novel type of probiotic chocolate with the additive Bacillus coagulans bacteria and determine the concentration of polyphenols and their bioaccessibility. The manufactured chocolate possessed significantly higher concentrations of polyphenols than the control sample. The sensory profiles of the tested probiotic chocolate were similar to those of the control sample. In future, the probiotic chocolate could be regarded as a functional food product by chocolate producers.

Protective Effects of Benzoic Acid, Bacillus Coagulans, and Oregano Oil on Intestinal Injury Caused by Enterotoxigenic Escherichia coli in Weaned Piglets.

The use of antibiotics as growth promoters in feed has been fully or partially banned in several countries. The objective of this study was to investigate the effects of benzoic acid (A), bacillus coagulans (B) and oregano oil (O) combined supplementation on growth performance and intestinal barrier in piglets challenged with enterotoxigenic Escherichia coli (ETEC). Thirty piglets were randomly assigned to 6 treatments: (1) nonchallenged control (CON); (2) ETEC-challenged control (ETEC); (3) antibiotics + ETEC (AT); (4) A + B + ETEC (AB); (5) A + O + ETEC (AO); (6) A + B + O + ETEC (ABO). On day 22, piglets were orally challenged with ETEC or saline. The trial lasted 26 days. Dietary AO and ABO inhibited the reduction of growth performance and the elevation of diarrhoea incidence in piglets induced by ETEC (P<0.05). AB, AO, and ABO prevented the elevation of serum TNF-α and LPS concentrations in piglets induced by ETEC (P<0.05). ABO alleviated the elevation of TNF-α and IL-1ß concentrations and the reduction of sIgA level in jejunal mucosa induced by ETEC (P<0.05). Furthermore, ABO upregulated mRNA expressions of Claudin-1 and Mucin2 (P<0.05), downregulated mRNA abundances of TLR4 and NOD2 signaling pathways related genes in jejunal mucosa (P<0.05), and improved the microbiota in jejunal and cecal digesta (P<0.05) compared with ETEC group. These results indicated that benzoic acid, bacillus coagulans, and oregano oil combined supplementation could improve growth performance and alleviate diarrhoea of piglets challenged with ETEC via improving intestinal mucosal barrier integrity, which was possibly associated with the improvement of intestinal microbiota and immune status. The combination of 3000 g/t benzoic acid + 400 g/t bacillus coagulans + 400 g/t oregano oil showed better effects than other treatments in improving growth performance and intestinal health of piglets, which could be used as a viable substitute for antibiotic.

Antiproliferative Effects of Bacillus coagulans Unique IS2 in Colon Cancer Cells.

In the present study, the in vitro anticancer (antiproliferative) effects of Bacillus coagulans Unique IS2 were evaluated on human colon cancer (COLO 205), cervical cancer (HeLa), and chronic myeloid leukemia (K562) cell lines with a human embryonic kidney cell line (HEK 293T) as noncancerous control cells. The Cytotoxicity assay (MTT) clearly demonstrated a 22%, 31.7%, and 19.5% decrease in cell proliferation of COLO 205, HeLa, and K562 cells, respectively, when compared to the noncancerous HEK 293T cells. Normal phase-contrast microscopic images clearly suggested that the mechanism of cell death is by apoptosis. To further confirm the induction of apoptosis by Unique IS2, the sub-G0-G1 peak of the cell cycle was quantified using a flow cytometer and the data indicated 40% of the apoptotic cells in Unique IS2-treated COLO cells when compared with their untreated control cells. The Western blot analysis showed an increase in pro-apoptotic protein BAX, decrease in antiapoptotic protein, Bcl2, decrease in mitochondrial membrane potential, increase in cytochrome c release, increase in Caspase 3 activity, and cleavage of poly(ADP-ribose) polymerase. The present study suggests that the heat-killed culture supernatant of B. coagulans can be more effective in inducing apoptosis of colon cancer cells and that can be considered for adjuvant therapy in the treatment of colon carcinoma.

Protective effects of synbiotic diets of Bacillus coagulans, Lactobacillus plantarum and inulin against acute cadmium toxicity in rats.

BACKGROUND: Cadmium is a heavy metal that causes oxidative stress and has toxic effects in humans. The aim of this study was to investigate the influence of two probiotics along with a prebiotic in preventing the toxic effects of cadmium in rats. METHODS: Twenty-four male Wistar rats were randomly divided into four groups namely control, cadmium only, cadmium along with Lactobacillus plantarum (1 × 109 CFU/day) and inulin (5% of feedstuff) and cadmium along with Bacillus coagulans (1 × 109 spore/day) and inulin (5% of feedstuff). Cadmium treated groups received 200 µg/rat/day CdCl2 administered by gavage. During the 42-day experimental period, they were weighed weekly. For evaluation of changes in oxidative stress, the levels of some biochemicals and enzymes of serum including SOD, GPX, MDA, AST, ALT, total bilirubin, BUN and creatinine, and also SOD level of livers were measured at day 21 and 42 of treatment. The cadmium content of kidney and liver was determined by using atomic absorption mass spectrophotometry. Data were analyzed using analysis of variance (ANOVA) followed by Duncan's post hoc test. RESULTS: Treatment of cadmium induced rats with synbiotic diets significantly improved the liver enzymes and biochemical parameters that decreased AST, ALT, total bilirubin, BUN and metal accumulation in the liver and kidney and increased body weight, serum and liver SOD values in comparison with the cadmium-treated group. No significant differences were observed with MDA and GPX values between all groups (p > 0.05). CONCLUSIONS: This study showed that synbiotic diets containing probiotics (L. plantarum and B. coagulans) in combination with the prebiotic (inulin) can reduce the level of cadmium in the liver and kidney, preventing their damage and recover antioxidant enzymes in acute cadmium poisoning in rat.

An In Vitro Approach to Study Effects of Prebiotics and Probiotics on the Faecal Microbiota and Selected Immune Parameters Relevant to the Elderly.

The aging process leads to alterations of gut microbiota and modifications to the immune response, such changes may be associated with increased disease risk. Prebiotics and probiotics can modulate microbiome changes induced by aging; however, their effects have not been directly compared. The aim of this study was to use anaerobic batch culture fermenters to assess the impact of various fermentable carbohydrates and microorganisms on the gut microbiota and selected immune markers. Elderly volunteers were used as donors for these experiments to enable relevance to an aging population. The impact of fermentation supernatants on immune markers relevant to the elderly were assessed in vitro. Levels of IL-1ß, IL-6, IL-8, IL-10 and TNF-α in peripheral blood mononuclear cell culture supernatants were measured using flow cytometry. Trans-galactooligosaccharides (B-GOS) and inulin both stimulated bifidobacteria compared to other treatments (p<0.05). Fermentation supernatants taken from faecal batch cultures supplemented with B-GOS, inulin, B. bifidum, L. acidophilus and Ba. coagulans inhibited LPS induced TNF-α (p<0.05). IL-10 production, induced by LPS, was enhanced by fermentation supernatants from faecal batch cultures supplemented with B-GOS, inulin, B. bifidum, L. acidophilus, Ba. coagulans and Bac. thetaiotaomicron (p<0.05). To conclude, prebiotics and probiotics could lead to potentially beneficial effects to host health by targeting specific bacterial groups, increasing saccharolytic fermentation and decreasing inflammation associated with aging. Compared to probiotics, prebiotics led to greater microbiota modulation at the genus level within the fermenters.

The Consumption of Synbiotic Bread Containing Lactobacillus sporogenes and Inulin Affects Nitric Oxide and Malondialdehyde in Patients with Type 2 Diabetes Mellitus: Randomized, Double-Blind, Placebo-Controlled Trial.

OBJECTIVES: To our knowledge, no reports are available indicating the effects of synbiotic bread consumption on nitric oxide (NO), biomarkers of oxidative stress, and liver enzymes among patients with type 2 diabetes mellitus (T2DM). This study was performed to determine the effects of the daily consumption of synbiotic bread on NO, biomarkers of oxidative stress, and liver enzymes in patients with T2DM. METHODS: This randomized, double-blind, placebo-controlled trial was performed among 81 patients with diabetes, aged 35-70 years old. After a 2-week run-in period, patients were randomly divided into 3 groups: group A (n = 27) received synbiotic bread containing viable and the heat-resistant probiotic Lactobacillus sporogenes (1 × 108 CFU) and 0.07 g inulin per 1 g, group B (n = 27) received probiotic bread containing Lactobacillus sporogenes (1 × 108 CFU), and group C (n = 27) received control bread for 8 weeks. Patients were asked to consume the synbiotic, probiotic, or control breads 3 times a day in 40 g packages for a total of 120 g/day. Fasting blood samples were taken at baseline and after an 8-week intervention for quantificationof related markers. RESULTS: After 8 weeks, the consumption of synbiotic bread compared to the probiotic and control breads resulted in a significant rise in plasma NO (40.6 ± 34.4 vs 18.5 ± 36.2 and -0.8 ± 24.5 µmol/L, respectively, p < 0.001) and a significant reduction in malondialdehyde (MDA) levels (-0.7 ± 0.7 vs 0.6 ± 1.7 and 0.5 ± 1.5 µmol/L, respectively, p = 0.001). We did not find any significant effect of the synbiotic bread consumption on plasma total antioxidant capacity (TAC), plasma glutathione (GSH), catalase, serum liver enzymes, calcium, iron, magnesium levels, and blood pressure compared to the probiotic and control breads. CONCLUSION: In conclusion, consumption of the synbiotic bread for 8 weeks among patients with T2DM had beneficial effects on plasma NO and MDA levels; however, it did not affect plasma TAC, GSH, catalase levels, serum liver enzymes, calcium, iron, magnesium levels, and blood pressure.