Animal Gut Health Research Articles

Isolation of swine-derived Lactobacillus plantarum and its synergistic antimicrobial and health-promoting properties with ZnO nanoparticles.

The purpose of this study was to isolate Lactobacillus from gastrointestinal tract of healthy postweaning piglets and investigate its synergistic antimicrobial and probiotic effects with ZnO nanoparticles (nZnO). Of the 128 isolates, Lactobacillus plantarum BLPL03 was selected based on its excellent acid and bile salt tolerance properties. Lactobacillus plantarum BLPL03 was sensitive to β-lactams, macrolides, amphenicols and cephalosporins, whereas it displayed the steady resistance to aminoglycosides, tetracyclines, quinolones and peptide antibiotics. In vitro analysis of antibacterial activities showed that L. plantarum BLPL03 inhibited the four common food-borne pathogenic bacteria including Escherichia coli O157:H7 CMCC 44828, Salmonella Typhimurium ATCC 13311, Staphylococcus aureus CMCC 26003 and Listeria monocytogenes CMCC 54007 in synergy with nZnO. Furthermore, the quantitative polymerase chain reaction test demonstrated that the combined administration of L. plantarum BLPL03 fermentation liquor (LFL) and nZnO synergistically elevated the faecal number of Bifidobacterium by 73·19-fold, and reduced the two potential enteropathogenic bacteria Enterobacteriaceae and Clostridium perfringens in mice challenged with Salm. Typhimurium. Finally, dietary supplementation with low dose of nZnO (20mgkg-1 ) when combined with LFL administration enhanced final body weight, fur appearance and average daily gain, and decreased feed conversion ratio and diarrhoea incidence in weaned piglets. The faecal Bifidobacterium and Lactobacillus of piglets were dramatically enhanced by 81·96- and 3·15-fold, respectively, after administration of a mixture of nZnO and LFL. Meanwhile, combination of nZnO with LFL resulted in low levels of Bacteroides, Enterococcus, and Enterobacteriaceae. A combination of nZnO and LFL exhibits potential health-benefit properties for the control of gut microbial composition by their synergistic antimicrobial and probiotic effects. This study may provide a potential nutritional strategy to improve performance and gut health of animals with gut microbiota disorders caused by pathogen infections and weanling, and so on.

Role of acidifiers in livestock nutrition and health: A review.

Ever since the European ban on use of in-feed antibiotics in food animals, the search for alternate antibiotic-free growth promoter is undertaken worldwide. There are few alternatives such as probiotics, pre-biotics, phytochemicals, enzymes and organic acids. Among these alternatives, the organic acids or simply acidifiers play an important role in gut health in animals. The acidifiers could be used to favourably manipulate the intestinal microbial populations and improve the immune response, hence perform an activity similar to antibiotics in food animals in countering pathogenic bacteria. Acidifiers also improve the digestibility of nutrients and increase the absorption of minerals. The incorporation of organic acids also leads to thinning of the intestinal lining which facilitates better absorption of nutrients and its efficient utilization. However, their effect will not be similar among all types of organic acids as their mechanism of activity is based on its pKa value. Moreover, there are claims about the neutralization of acids by the secretion of bicarbonates in the initial part of intestine, reactivity with metallic items in feed mills and reduced palatability due its bitter taste demands non-reactive and targeted delivery for better performance. Currently, coated salts of acidifiers are available commercially for use in food animals especially pigs and poultry. The present review highlights the role of different acidifiers in livestock nutrition with their potent applications in improving nutrient digestibility, mineral utilization, meat quality, enhancing immunity, antimicrobial effects in countering pathogenic bacteria, boosting performance and production, and thus safeguarding health of livestock animals and poultry.

Comprehensive Understanding of the Bacterial Populations and Metabolites Profile of Fermented Feed by 16S rRNA Gene Sequencing and Liquid Chromatography-Mass Spectrometry.

The comprehensive bacterial populations and metabolites profile in fermented feed is unclear, which may have significant effects on the stability of fermented feed quality and animal gut health. In this study, 16S rRNA gene sequencing and liquid chromatography–mass spectrometry were used to explore the bacterial populations and metabolites profile in the fermented feed incubated with probiotics (MF) or without probiotics (SF). The probiotics were a combination of Lactobacillus salivarius, Bacillus subtilis, and Saccharomyces cerevisiae. The pH and lactic acid levels were higher in MF than in SF (P < 0.05), while the total volatile fatty acid content was lower (P < 0.05). Interestingly, after fermentation, the most abundant bacterial genus in MF was Enterococcus, rather than the added probiotics Lactobacillus or Bacillus. Weissella and a few potential pathogens (Enterobacter, Escherichia-Shigella, and Pantoea) were dominant in SF (P < 0.05). Metabolomics analysis identified 32 different metabolites in the two types of fermented feed. These metabolites enriched in MF, such as maleic acid, phenylacetic acid, ethyl linoleate, dihomo-gamma-linolenic acid, and L-theanine had potential antimicrobial activities. Conclusively, the addition of probiotics enriched a few potentially beneficial microbes and small molecular compounds with antimicrobial activities, and inhibited the potential pathogens in fermented feed.

Succinate Modulates Intestinal Barrier Function and Inflammation Response in Pigs.

Succinate is a metabolic intermediate of the tricarboxylic acid (TCA) cycle in all aerobic organisms, and is also a vital microbial metabolite in the gut. Although succinate is known to regulate intestinal metabolism and immune function, its role in the protection of the intestinal epithelial barrier function and inflammation is poorly characterized. In this study, we evaluated the effects of succinate on intestinal epithelial barrier function and inflammation in pigs. Twenty-four growing pigs were distributed into three groups (n = 8) and received either a basal diet (control group) or the same diet supplemented with 0.1% succinate or 1% succinate. The diet supplemented with 1% succinate led to alterations in the intestinal morphology. We confirmed in vitro that 5 mM succinate treatment modulated intestinal epithelial permeability by increased transepithelial electrical resistance (TEER) in intestinal porcine epithelial cell (IPEC)-J2 cells. Furthermore, succinate treatment increased the abundance of tight junction proteins claudin-1, zona occluden (ZO)-1, and ZO-2 in the jejunum in vivo and in vitro. In addition, dietary succinate supplementation promoted the expression of inflammatory cytokines interleukin (IL)-25, IL-10, IL-8, and IL-18 in the jejunum. Taken together, these data identify a novel role of succinate in the modulation of intestinal epithelial barrier function, which may be a nutritional target to improve gut health in animals.

Early Nutrition Programming (in ovo and Post-hatch Feeding) as a Strategy to Modulate Gut Health of Poultry.

Healthy gastrointestinal tract (GIT) is crucial for optimum performance, better feed efficiency, and overall health of poultry. In the past, antibiotic growth promoters (AGP) were commonly used to modulate the gut health of animals. However, considering the public health concern, the use of AGP in animal feeding is banned or regulated in several jurisdictions around the world. This necessitates the need for alternative nutritional strategies to produce healthy poultry. For that, several alternatives to AGP have been attempted with some success. However, effective modulation of the gut health parameters depends on the methods and timing of the compound being available to host animals. Routinely, the alternatives to AGP and other nutrients are provided in feed or water to poultry. However, the GIT of the newly hatched poultry is functionally immature, despite going through significant morphological, cellular, and molecular changes toward the end of incubation. Thus, early growth and development of GIT are of critical importance to enhance nutrients utilization and optimize the growth of poultry. Early nutrition programming using both in ovo and post-hatch feeding has been used as a means to modulate the early growth and development of GIT and found to be an effective strategy but with inconsistent results. This review summarizes the information on in ovo and post-hatch-feeding of different nutrients and feeds additives and their effects on gut development, histomorphology, microbiology, and immunology. Furthermore, this review will provide insight on the future of early nutrition programming as a strategy to enhance gut health, thereby improving overall health and production so that the poultry industry can benefit from this technique.

Dietary Lysozyme Alters Sow’s Gut Microbiota, Serum Immunity and Milk Metabolite Profile

The aim of current study was to determine variations in sow’s gut microbiota, serum immunity, and milk metabolite profile mediated by lysozyme supplementation. Twenty-four pregnant sows were assigned to a control group without supplementation and two treatments with 0.5 kg/t and 1.0 kg/t lysozyme provided in formula feed for 21 days (n = 8 per treatment). Microbiota analysis and metagenomic predictions were based on 16s RNA high-throughput sequencing. Milk metabolome was assessed by untargeted liquid chromatography tandem mass spectrometry. Serum biochemical indicators and immunoglobulins were also determined. Gut microbial diversity of sows receiving 1.0 kg/t lysozyme treatment was significantly reduced after the trial. Spirochaetes, Euryarchaeota, and Actinobacteria significantly increased while Firmicutes showed a remarkable reduction in 1.0 kg/t group compared with control. Lysozyme addition rebuilt sow’s gut microbiota to beneficial composition identified by reduced richness of Escherichia coli and increased abundance of Lactobacillus amylovorus. Accordingly, microbial metabolic functions including pyrimidine metabolism, purine metabolism, and amino acid related enzymes were significantly up-regulated in 1.0 kg/t group. Microbial metabolic phenotypes like the richness of Gram-positive bacteria and oxidative stress tolerance were also significantly reduced by lysozyme treatment. Serum alanine transaminase (ALT) activity and IgA levels were significantly down-regulated in the 1.0 kg/t group compared with control, but IgM levels showed a significantly increase in 1.0 kg/t group. Milk metabolites such as L-glutamine, creatine, and L-arginine showed significantly dose-dependent changes after treatment. Overall, lysozyme supplementation could effectively improve the composition, metabolic functions, and phenotypes of sow’s gut microbiota and it also benefit sows with better serum immunity and milk composition. This research could provide theoretical support for further application of lysozyme in promoting animal gut health and prevent pathogenic infections in livestock production.

Effects of dietary Bacillus amyloliquefaciens supplementation on growth performance, intestinal morphology, inflammatory response, and microbiota of intra-uterine growth retarded weanling piglets

BackgroundThe focus of recent research has been directed toward the probiotic potential of Bacillus amyloliquefaciens (BA) on the gut health of animals. However, little is known about BA’s effects on piglets with intra-uterine growth retardation (IUGR). Therefore, this study investigated the effects of BA supplementation on the growth performance, intestinal morphology, inflammatory response, and microbiota of IUGR piglets.MethodsEighteen litters of newborn piglets were selected at birth, with one normal birth weight (NBW) and two IUGR piglets in each litter (i.e., 18 NBW and 36 IUGR piglets in total). At weaning, the NBW piglet and one of the IUGR piglets were assigned to groups fed a control diet (i.e., the NBW-CON and IUGR-CON groups). The other IUGR piglet was assigned to a group fed the control diet supplemented with 2.0 g BA per kg of diet (i.e., IUGR-BA group). The piglets were thus distributed across three groups for a four-week period.ResultsIUGR reduced the growth performance of the IUGR-CON piglets compared with the NBW-CON piglets. It was also associated with decreased villus sizes, increased apoptosis rates, reduced goblet cell numbers, and an imbalance between pro- and anti-inflammatory cytokines in the small intestine. Supplementation with BA improved the average daily weight gain and the feed efficiency of the IUGR-BA group compared with the IUGR-CON group (P < 0.05). The IUGR-BA group exhibited increases in the ratio of jejunal villus height to crypt depth, in ileal villus height, and in ileal goblet cell density. They also exhibited decreases in the numbers of jejunal and ileal apoptotic cells and ileal proliferative cells (P < 0.05). Supplementation with BA increased interleukin 10 content, but it decreased tumor necrosis factor alpha level in the small intestines of the IUGR-BA piglets (P < 0.05). Furthermore, compared with the IUGR-CON piglets, the IUGR-BA piglets had less Escherichia coli in their jejunal digesta, but more Lactobacillus and Bifidobacterium in their ileal digesta (P < 0.05).ConclusionsDietary supplementation with BA improves morphology, decreases inflammatory response, and regulates microbiota in the small intestines of IUGR piglets, which may contribute to improved growth performance during early life.

Implications of butyrate and its derivatives for gut health and animal production.

Butyrate is produced by microbial fermentation in the large intestine of humans and animals. It serves as not only a primary nutrient that provides energy to colonocytes, but also a cellular mediator regulating multiple functions of gut cells and beyond, including gene expression, cell differentiation, gut tissue development, immune modulation, oxidative stress reduction, and diarrhea control. Although there are a large number of studies in human medicine using butyrate to treat intestinal disease, the importance of butyrate in maintaining gut health has also attracted significant research attention to its application for animal production, particularly as an alternative to in-feed antibiotics. Due to the difficulties of using butyrate in practice (i.e., offensive odor and absorption in the upper gut), different forms of butyrate, such as sodium butyrate and butyrate glycerides, have been developed and examined for their effects on gut health and growth performance across different species. Butyrate and its derivatives generally demonstrate positive effects on animal production, including enhancement of gut development, control of enteric pathogens, reduction of inflammation, improvement of growth performance (including carcass composition), and modulation of gut microbiota. These benefits are more evident in young animals, and variations in the results have been reported. The present article has critically reviewed recent findings in animal research on butyrate and its derivatives in regard to their effects and mechanisms behind and discussed the implications of these findings for improving animal gut health and production. In addition, significant findings of medical research in humans that are relevant to animal production have been cited.

The mycotoxins deoxynivalenol and nivalenol show in vivo synergism on jejunum enterocytes apoptosis

The mycotoxins deoxynivalenol (DON) and nivalenol (NIV), worldwide cereal contaminants, raise concerns for human and animal gut health, following exposure through contaminated food and feed. The aim of this work was to analyze the effects of DON and NIV, alone or associated, on the intestinal pig mucosa. Jejunal loops were used for testing DON and NIV individually and in combination (1:1) after a single exposure, for 24 h. For repeated exposure, piglets received a natural contaminated feed, with DON or with DON + NIV for 28 days. Histological investigations, proliferation and apoptosis assessments were conducted. Both experiments were concordant for the total-cell proliferation decreased at the villus tips after DON or DON + NIV at 10 μM acutely, or repeatedly, by 30–35% and 20–25%, respectively. In loops model, apoptotic enterocytes at villus tips increased dose-dependently after DON, NIV alone or DON + NIV in combination. The combination in loops at 10 μM showed higher effects on proliferation and apoptosis than DON alone, and synergism was shown for villus apoptotic enterocyte. These results are to be considered for NIV consumer risk assessment. Our results demonstrate the in vivo disruption of the intestinal balance proliferation/apoptosis explaining, at least partly, the disruption of intestinal barrier by these mycotoxins.

Regulation of the intestinal immune system by commensal microbiota in avian and ruminant species.

Abstract Maintenance of intestinal immune system homeostasis requires the host to distinguish commensal bacteria, which are members of the gut microbiota, from pathogenic micro-organisms and respond accordingly. Although the mechanisms that maintain this homeostasis are not completely understood, evidence, particularly from studies of rodents and humans, has enabled the unravelling of the finely tuned balance that exists between the host and its microbiota. Commensal bacteria, which are in close contact with cells of the intestinal immune system, have been shown to play an important role in regulation of innate and adaptive immune mechanisms. The anatomy of the gut-associated lymphoid tissue in avian and ruminant species differs from that of monogastric animal species. However, many of the mechanisms that are used to identify and respond to commensal bacteria are highly conserved among species and are, therefore, thought to function similarly. In recent years, interest in the area of gut microbiota interactions with the host immune system has significantly increased. This is partly because of the implications of these interactions for the development of novel strategies for improving gut health in animals.

Effects of lactoferrin supplementation on ileal and total tract nutrient digestibility, gastrointestinal microbial populations, and immune characteristics of ileal cannulated, healthy, adult dogs

Orally supplemented lactoferrin derived from bovine milk is purported to have beneficial effects on gut health of animals. Bovine lactoferrin (0, 60, or 120 mg/d) was fed to ileal cannulated, adult dogs in a replicated 3 × 3 Latin square design with 14 d periods. Control dogs tended (p = 0.06) to have higher fecal DM concentrations compared with dogs supplemented with 120 mg/d lactoferrin (34.5 vs. 32.9%). Fecal scores ranged from 3.0 – 3.3, suggesting that feces of all dogs was near the desired consistency, with dogs supplemented with 120 mg/d lactoferrin tending (p = 0.08) to have higher fecal scores. Ileal azoreductase activity tended (p < 0.10) to be higher in dogs supplemented with 60 or 120 mg/d lactoferrin (609 vs. 592 nmol/h per g ileal DM, respectively) as compared with unsupplemented dogs (272 nmol/h per g ileal DM). The following bacterial groups were measured: bifidobacteria, Campylobacter spp., Clostridium spp., eubacteria, Escherichia coli, Lactobacillus spp., Staphylococcus spp., and Streptococcus spp. Fecal streptococci concentrations were lower (p < 0.05) for dogs receiving 60 mg/d lactoferrin (8.60 log10 cfu/g fecal DM) as compared with unsupplemented dogs (9.19 log10 cfu/g fecal DM) or dogs receiving 120 mg lactoferrin/d (9.43 log10 cfu/g fecal DM). Dogs supplemented with 120 mg/d lactoferrin tended (p = 0.08) to have higher fecal indole concentrations as compared to unsupplemented dogs (1.80 vs. 1.46 μmol/g fecal DM). Because most bacterial groups measured were unaffected, it appears that lactoferrin did not exhibit prebiotic activity, and based on the data collected, lactoferrin also did not appear to have major effects on indices of health in the dog.