The probiotic fermented milk of Lacticaseibacillus paracasei JY062 and Lactobacillus gasseri JM1 alleviates constipation via improving gastrointestinal motility and gut microbiota.

Constipation is directly related to the intestinal microenvironment, in which the promotion of gastrointestinal (GI) motility and improvement of gut microbiota distribution are important for alleviating symptoms. Herein, after the intervention of probiotic fermented milk (FMMIX) containing Lacticaseibacillus paracasei JY062 and Lactobacillus gasseri JM1 for 14 d in Kunming mice with loperamide-induced constipation, the results indicated that FMMIX significantly increased the secretion of serum motilin, gastrin and 5-hydroxytryptamine, as well as decreased the secretion of peptide YY, vasoactive intestinal peptide, and nitric oxide in mice. As determined by immunohistochemical analysis, FMMIX promoted an augmentation in the quantity of Cajal interstitial cells. In addition, the mRNA and protein expression of c-kit and stem cell factor (SCF) were upregulated to facilitate intestinal motility. High-throughput sequencing and gas chromatography techniques revealed that FMMIX led to an increase in the relative abundance of beneficial bacteria (Lactobacillus, Oscillospira, Ruminococcus, Coprococcus, and Akkermansia), reduced the presence of harmful bacteria (Prevotella), and resulted in elevated levels of short-chain fatty acids (SCFA) with a superior improvement compared with unfermented milk. Untargeted metabolomics revealed significant upregulation of functional metabolites such as l-pipecolinic acid, dl-phenylalanine, and naringenin in FMMIX, presumably playing a potential role in constipation relief. Overall, our results showed that FMMIX had the potential to alleviate constipation symptoms in mice by improving the secretion of serum GI regulatory peptides and neurotransmitters, increasing the expression of c-kit and SCF proteins, and modulating the gut microbiota structure and SCFA levels, and may be associated with an increase in these functional metabolites. This suggested that FMMIX could be a promising adjunctive strategy for managing constipation symptoms and could contribute to the development of functional foods aimed at improving gut health.

Modulation of growth performance, antioxidant capacity, non-specific immunity and disease resistance in largemouth bass (Micropterus salmoides) upon compound probiotic cultures inclusion.

An 8-week feeding trial was conducted to evaluate the effects of dietary supplementation of compound probiotic cultures (CPC; Bacillus subtilis, Lactobacillus plantarum and Saccharomyces cerevisiae) on the growth performance, antioxidant capacity, non-specific immunity and disease resistance of juvenile largemouth bass. Triplicate groups of largemouth bass (average weight 42.05 ± 0.02 g), with a destiny of 30 individuals per tank, were fed diets supplemented with different concentration of compound probiotic cultures (CPC) (0%, CPC (0.0); 0.5%, CPC (0.5); 1.0%, CPC (1.0); 2.0%, CPC (2.0)). After the feeding trial, tissue samples of largemouth bass were collected and the challenge test with Aeromonas hydrophila was performed. Results indicated that the CPC supplementation produced no significant difference on the growth performance, feed utilization and body composition of largemouth bass, while significantly increased the cumulative survival rate in the Aeromonas hydrophila challenge test. Meanwhile, the inclusion of CPC elevated the hepatic antioxidant capacity, and the highest activity of antioxidant enzymes, including T-AOC, CAT, GPx and T-SOD, was observed in the CPC (2.0) group. Meanwhile, the transcription of Nrf2/keap1 and antioxidant related genes, including CAT, GPx, GST, SOD1 and SOD2, was significantly elevated with the inclusion of CPC. In addition, the inclusion of CPC improved the non-specific immunity of largemouth bass. The activity of serum lysozyme was significantly elevated in the CPC (2.0) group, while the transcription of RelA and pro-inflammatory factors, including TNF-α and IL-1ß, was inhibited with the inclusion of CPC. Meanwhile, related genes potentially linked to RelA, including TLR2 and p38 MAPK, were detected that their relative expression was significantly inhibited with the inclusion of CPC. The current findings indicated that the inclusion of 2% CPC improved the antioxidant capacity, non-specific immunity and disease resistance of juvenile largemouth bass, and suggested that 2% CPC as a functional additive could be applied to the diet of juvenile largemouth bass in aquaculture practice.

Effects of compound probiotics on growth performance, rumen fermentation, blood parameters, and health status of neonatal Holstein calves.

This study aimed to investigate the effects of compound probiotics (consisting of 108 cfu/g of Lactobacillus plantarum, 108 cfu/g of Pediococcus acidilactici, 108 cfu/g of Pediococcus pentosaceus, 107 cfu/g of and Bacillus subtilis) on growth performance, rumen fermentation, bacteria community, blood parameters, and health status of Holstein calves at the first 3 mo of age. Forty-eight newborn calves were randomly divided into the following 3 groups: control group (milk replacer with no compound probiotics), low compound probiotics group (milk replacer + 0.12 g of compound probiotics per head per day), and high compound probiotics group (HP; milk replacer + 1.2 g of compound probiotics per head per day). Starter pellets of the low compound probiotics and HP groups were coated with 0.05% compound probiotics. Milk replacer was provided from 2 to 63 d of age (6 L at 2-10 d, 8 L at 11-42 d, 6 L at 43-49 d, 4 L at 50-56 d, and 2 L at 57-63 d), and starter pellets were provided ad libitum from 7 to 90 d of age. Body weight and body size (d 1, 30, 60, and 90), blood (d 40 and 80), and rumen fluid (d 90) were analyzed using the one-way ANOVA procedure; fecal score was recorded daily and analyzed as repeated measures using the mixed model procedure. Results showed that diet supplemented with compound probiotics had no effects on the body weight, average daily gain, dry matter intake, and feed efficiency. At 90 d of age, diet supplemented with compound probiotics decreased the withers height. Immunity activities increased in the HP group, supported by the increased concentrations of serum total protein and immunoglobulins at 40 d of age, and by the increased activity of superoxide dismutase at 80 d of age. Diet supplemented with compound probiotics altered rumen fermentation, indicated by the decreased rumen acetic acid and propionic acid, and the increased butyric acid concentrations. Diet supplemented with compound probiotics improved the health status of calves, indicated by the decreased fecal score at 3 wk of age and the decreased medicine treatments. In summary, although diet supplemented with HP decreased the withers height, this level of probiotics is recommended to improve rumen development and health status of newborn Holstein calves.

Spraying compound probiotics improves growth performance and immunity and modulates gut microbiota and blood metabolites of suckling piglets.

One factor that shapes the establishment of early neonatal intestinal microbiota is environmental microbial exposure, and probiotic application has been shown to promote health and growth of piglets. Thus, this study hypothesized that environmental probiotic application in early days of life would be beneficial to newborn piglets. This study aimed to investigate the effect of spraying a compound probiotic fermented liquid (CPFL) into the living environment of piglets on their early growth performance and immunity. This work included 68 piglets, which were randomized into probiotic and control groups. Blood and fecal samples were collected at 0, 3, 7, 14, and 21 days of age. Spraying CPFL significantly reshaped the microbiota composition of the delivery room environment, increased piglets' daily weight gain and weaning weight (P<0.001), and modulated piglets' serum cytokine levels (increases in IgA, IgG, and IL-10; decrease in IFN-γ; P<0.05 in each case) in piglets. Additionally, spraying CPFL during early days of life modified piglets' gut microbiota structure and diversity, increased the abundance of some potentially beneficial bacteria (such as Bacteroides uniformis, Butyricimonas virosa, Parabacteroides distasonis, and Phascolarctobacterium succinatutens) and decreased the abundance of Escherichia coli (P<0.05). Interestingly, CPFL application also significantly enhanced the gut microbial bioactive potential and levels of several serum metabolites involved in the metabolism of vitamins (B2, B3, B6, and E), medium/long-chain fatty acids (caproic, tetradecanoic, and peptadecanoic acids), and dicarboxylic acids (azelaic and sebacic acids). Our study demonstrated that spraying CPFL significantly could improve piglets' growth performance and immunity, and the beneficial effects are associated with changes in the gut microbiota and host metabolism. Our study has provided novel data for future development of probiotic-based health-promoting strategies and expanded our knowledge of probiotic application in animal husbandry.

Compound Probiotic Ameliorates Acute Alcoholic Liver Disease in Mice by Modulating Gut Microbiota and Maintaining Intestinal Barrier.

Alcoholic liver disease (ALD) is a worldwide health threaten lack of effective treatment. Gut dysbiosis and concomitant augmented intestinal permeability are strongly implicated in the pathogenesis and progression of ALD. Research on the protective effect of probiotics on ALD is limited, and more effective intestinal microecological regulators and the related mechanisms still need to be further explored. In the present study, the protective effects and mechanisms of a compound probiotic against acute alcohol-induced liver injury in vivo were explod. It was showed that the compound probiotic ameliorated liver injury in acute ALD mice and stabilized the levels of ALT, AST, and TG in serum. The compound probiotic reversed acute alcohol-induced gut dysbiosis and maintained the intestinal barrier integrity by upregulating the production of mucus and the expression of tight junction (TJ) proteins and thus reduced LPS level in liver. Meanwhile, the compound probiotic reduced inflammation level by inhibiting TLR4/NF-κB signaling pathway and suppressed oxidative stress level in liver. Furthermore, the compound probiotic alleviated liver lipid accumulation by regulating fatty acid metabolism-associated genes and AMPK-PPARα signaling pathway. Noteworthy, fecal microbiota transplantation (FMT) realized comparable protective effect with that of compound probiotic. In conclusion, present study demonstrates the beneficial effects and underlying mechanism of the compound probiotic against acute alcohol-induced liver injury. It provides clues for development of novel strategy for treatment of ALD.

Probiotics alleviate autoimmune hepatitis in mice through modulation of gut microbiota and intestinal permeability.

Autoimmune hepatitis (AIH) is an immune-mediated type of chronic liver inflammation accompanied by intestinal flora imbalance. Probiotics have been reported to ameliorate imbalances in the intestinal flora. This study aimed to investigate the effects of compound probiotic in the AIH mouse model. AIH mice were gavaged with compound probiotic and injected intraperitoneally with dexamethasone (dex) for 42 days. The results showed that these treatments suppressed hepatic inflammatory cell infiltration, serum transaminase, and Th1 and Th17 cells. However, Treg cells were increased only in the probiotics group, which indicates an immunomodulatory role of the compound probiotic. The compound probiotic maintained intestinal barrier integrity, blocked lipopolysaccharide (LPS) translocation, and inhibited the activation of the TLR4/NF-κB pathway and the production of inflammatory factors in the liver and ileum. Moreover, the compound probiotic treatment increased the abundance of beneficial bacteria and reduced the abundance of potentially harmful bacteria in gut. Compound probiotic may improve ileal barrier function while increasing the diversity of the intestinal flora, blocking the translocation of gut-derived LPS to the liver and therefore preventing activation of the TLR4/NF-κB pathway. The resulting inhibition of pro-inflammatory factor production facilitates AIH remission.