Fecal Bacterial Populations Research Articles

Impact of Lactobacillus- and Bifidobacterium-Based Direct-Fed Microbials on the Performance, Intestinal Morphology, and Fecal Bacterial Populations of Nursery Pigs.

Weaning is a critical stage in the swine production cycle, as young pigs need to adjust to sudden and dramatic changes in their diet and environment. Among the various organ systems affected, the gastrointestinal tract is one of the more severely impacted during this transition. Traditionally, challenges at weaning have been managed by prophylactic use of antibiotics, which not only provides protection against diarrhea and other gut dysfunction but also has growth-promoting effects. With banning or major restrictions on the use of antibiotics for this purpose, various alternative products have been developed as potential replacements, including direct-fed microbials (DFMs) such as probiotics and postbiotics. As their efficiency needs to be improved, a continued effort to gain a deeper understanding of their mechanism of action is necessary. In this context, this report presents a study on the impact of a Lactobacillus-based probiotic (LPr) and a Bifidobacterium-based postbiotic (BPo) when added to the diet during the nursery phase. For animal performance, an effect was observed in the early stages (Day 0 to Day 10), as pigs fed diets supplemented with either DFMs were found to have higher average daily feed intake (ADFI) compared to pigs fed the control diet (p < 0.05). Histological analysis of intestinal morphology on D10 revealed that the ileum of supplemented pigs had a higher villus height/crypt depth ratio (p < 0.05) compared to controls, indicating a benefit of the DFMs for gut health. In an effort to further explore potential mechanisms of action, the effects of the DFMs on gut microbial composition were investigated using fecal microbial communities as a non-invasive representative approach. At the bacterial family level, Lactobacillaceae were found in higher abundance in pigs fed either LPr (D10; p < 0.05) or BPo (D47; p < 0.05). At the Operational Taxonomic Unit (OTU) level, which can be used as a proxy to assess species composition, Ssd-00950 and Ssd-01187 were found in higher abundance in DFM-supplemented pigs on D47 (p < 0.05). Using nucleotide sequence identity, these OTUs were predicted to be putative strains of Congobacterium massiliense and Absicoccus porci, respectively. In contrast, OTU Ssd-00039, which was predicted to be a strain of Streptococcus alactolyticus, was in lower abundance in BPo-supplemented pigs on D47 (p < 0.05). Together, these results indicate that the DFMs tested in this study can impact various aspects of gut function.

Temporal dynamics of the fecal microbiome in female pigs from early life through estrus, parturition, and weaning of the first litter of piglets.

Age-associated changes in the gastrointestinal microbiome of young pigs have been robustly described; however, the temporal dynamics of the fecal microbiome of the female pig from early life to first parity are not well understood. Our objective was to describe microbiome and antimicrobial resistance dynamics of the fecal microbiome of breeding sows from early life through estrus, parturition and weaning of the first litter of piglets (i.e., from 3 to 53weeks of age). Our analysis revealed that fecal bacterial populations in developing gilts undergo changes consistent with major maturation milestones. As the pigs progressed towards first estrus, the fecal bacteriome shifted from Rikenellaceae RC9 gut group- and UCG-002-dominated enterotypes to Treponema- and Clostridium sensu stricto 1-dominated enterotypes. After first estrus, the fecal bacteriome stabilized, with minimal changes in enterotype transition and associated microbial diversity from estrus to parturition and subsequent weaning of first litter piglets. Unlike bacterial communities, fecal fungal communities exhibited low diversity with high inter- and intra-pig variability and an increased relative abundance of certain taxa at parturition, including Candida spp. Counts of resistant fecal bacteria also fluctuated over time, and were highest in early life and subsequently abated as the pigs progressed to adulthood. This study provides insights into how the fecal microbial community and antimicrobial resistance in female pigs change from three weeks of age throughout their first breeding lifetime. The fecal bacteriome enterotypes and diversity are found to be age-driven and established by the time of first estrus, with minimal changes observed during subsequent physiological stages, such as parturition and lactation, when compared to the earlier age-related shifts. The use of pigs as a model for humans is well-established, however, further studies are needed to understand how our results compare to the human microbiome dynamics. Our findings suggest that the fecal microbiome exhibited consistent changes across individual pigs and became more diverse with age, which is a beneficial characteristic for an animal model system.

Application of Compositional Data Analysis to Study the Relationship between Bacterial Diversity in Human Faeces and Sex, Age, and Weight.

This work uses Compositional Data Analysis (CoDA) to examine the typical human faecal bacterial diversity in 39 healthy volunteers from the Andalusian region (Spain). Stool samples were subjected to high-throughput sequencing of the V3 and V4 regions of the 16S ribosomal RNA gene using Illumina MiSeq. The numbers of sequences per sample and their genus-level assignment were carried out using the Phyloseq R package. The alpha diversity indices of the faecal bacterial population were not influenced by the volunteer's sex (male or female), age (19-46 years), and weight (48.6-99.0 kg). To study the relationship between these variables and the faecal bacterial population, the ALDEx2 and coda4microbiome CoDA packages were used. Applying ALDEx2, a trend suggesting a connection between sex and the genera Senegalimassilia and Negatibacillus (slightly more abundant in females) and Desulfovibrio (more abundant in males) was found. Moreover, age was tentatively associated with Streptococcus, Tizzerella, and Ruminococaceae_UCG-003, while weight was linked to Senegalimassilia. The exploratory tool of the coda4microbiome package revealed numerous bacterial log-ratios strongly related to sex and, to a lesser extent, age and weight. Moreover, the cross-sectional analysis identified bacterial signature balances able to assign sex to samples regardless of controlling for volunteers' age or weight. Desulfovibrio, Faecalitalea, and Romboutsia were relevant in the numerator, while Coprococcus, Streptococcus, and Negatibacillus were prominent in the denominator; the greater presence of these could characterise the female sex. Predictions for age included Caproiciproducens, Coprobacter, and Ruminoclostridium in the numerator and Odoribacter, Ezakiella, and Tyzzerella in the denominator. The predictions depend on the relationship between both groups, but the abundance of the first group and scarcity of the second could be related to older individuals. However, the association of the faecal bacterial population with weight did not yield a satisfactory model, indicating scarce influence. These results demonstrate the usefulness of the CoDA methodology for studying metagenomics data and, specifically, human microbiota.

Bile Acids: Major Regulator of the Gut Microbiome.

Bile acids are synthesized from cholesterol and play an important role in regulating intestinal microflora. The different degrees of hydrophobicity and acidity of individual bile acids may affect their antimicrobial properties. We examined the antimicrobial effects of different bile acids on various microorganisms in vitro and confirmed whether these remain consistent in vivo. Using human bile acids, including ursodeoxycholic acid, cholic acid, chenodeoxycholic acid, deoxycholic acid, and lithocholic acid, a disc diffusion test was performed, and a rodent model was created to determine the antimicrobial effects of each bile acid. The fecal bacterial population was analyzed using a real-time polymerase chain reaction. Each bile acid showed different microbial inhibitory properties. The inhibitory activity of bile acids against microbiota which normally resides in the gastrointestinal tract and biliary system, was low; however, normal flora of other organs was significantly inhibited. Changes in microbial counts after bile acid administration in a rodent model differed in the colon and cecum. The in vivo and in vitro results show that the antimicrobial effects of bile acids against intestinal microbiota were similar. In conclusion, bile acids could be a novel treatment strategy to regulate gut microbiota.

The Effects of Sodium Butyrate, Coated Sodium Butyrate, and Butyric Acid Glycerides on Nutrient Digestibility, Gastrointestinal Function, and Fecal Microbiota in Turkeys.

Simple SummaryThe development of antimicrobial resistance is one the most serious health threats. Therefore, alternatives to antibiotic growth promoters that could be used in animal production are sorely needed. Organic acids, including butyric acid, are among the most promising compounds. Butyrate exhibits antimicrobial activity; it decreases the pH of intestinal digesta and decreases the amounts of pathogenic microbes. In this study, turkeys were fed diets with various forms of butyric acid. The growth performance of turkeys and duodenal villus height increased, whereas the fecal populations of Escherichia coli and Clostridium perfringens decreased when butyric acid glycerides or coated sodium butyrate were added to turkey diets. An increase in protein digestibility and a decrease in the gizzard pH were noted in birds fed diets with butyric acid glycerides. The addition of butyric acid in different forms to diets increased the butyric acid concentration in the cecal digesta of turkeys. The study results suggest that protected forms of butyric acid improve growth performance and protein digestibility and decrease the fecal populations of pathogenic bacteria in turkeys. Therefore, protected forms of butyric acid can be valuable feed additives in turkey nutrition.This study aimed to determine the efficacy of sodium butyrate (SB), coated sodium butyrate (CSB), and butyric acid glycerides (BAG) in turkey nutrition based on an analysis of nutrient digestibility, gastrointestinal function, and fecal bacterial populations. A total of 400 1-day-old female BIG 6 turkeys were divided into 4 groups, with 5 replicates per group and 20 birds per replicate, to determine the effects exerted by various forms of butyric acid (SB, CSB, and BAG). The addition of CSB and BAG to turkey diets improved the feed conversion ratio (FCR, p < 0.05), increased the values of the European Efficiency Index (EEI, p < 0.01) and duodenal villus height (p < 0.05), and decreased the fecal populations of Escherichia coli and Clostridium perfringens (p < 0.05). Dietary supplementation with BAG increased protein digestibility (p < 0.05). The analyzed forms of butyrate added to turkey diets increased the butyric acid concentration in the cecal digesta (p < 0.01). The results of this study indicate that protected forms of butyric acid can be valuable feed additives in turkey nutrition.

Lower systemic inflammation is associated with gut firmicutes dominance and reduced liver injury in a novel ambulatory model of parenteral nutrition

Background Total Parenteral Nutrition (TPN) provides lifesaving nutritional support to patients unable to maintain regular enteral nutrition (EN). Unfortunately, cholestasis is a significant side effect affecting 20–40% of paediatric patients. While the aetiology of TPN-associated injury remains ill-defined, an altered enterohepatic circulation in the absence of gut luminal nutrient content during TPN results in major gut microbial clonal shifts, resulting in metabolic endotoxemia and systemic inflammation driving liver injury and cholestasis. Hypothesis To interrogate the role of gut microbiota, using our novel ambulatory TPN piglet model, we hypothesized that clonal reduction of bacteria in Firmicutes phylum (predominant in EN) and an increase in pathogenic Gram-negative bacteria during TPN correlates with an increase in serum lipopolysaccharide and systemic inflammatory cytokines, driving liver injury. Methods Upon institutional approval, 16 animals were allocated to receive either TPN (n = 7) or EN only (n = 9). The TPN group was subdivided into a low systemic inflammation (TPN-LSI) and high systemic inflammation (TPN-HSI) based on the level of serum lipopolysaccharide. Culture-independent identification of faecal bacterial populations was determined by 16S rRNA. Results Piglets on TPN, in the TPN-HSI group, noted a loss of enterocyte protective Firmicutes bacteria and clonal proliferation of potent inflammatory and lipopolysaccharide containing pathogens: Fusobacterium, Bacteroidetes and Campylobacter compared to EN animals. Within the TPN group, the proportion of Firmicutes phylum correlated with lower portal lipopolysaccharide levels (r = −0.89). The TPN-LSI had a significantly lower level of serum bile acids compared to the TPN-HSI group (7.3 vs. 60.4 mg/dL; p = .018), increased day 14 weight (5.67 vs. 5.07 kg; p = .017) as well as a 13.7-fold decrease in serum conjugated bilirubin. Conclusion We demonstrate a novel relationship between the gut microbiota and systemic inflammation in a TPN animal model. Pertinently, the degree of gut dysbiosis correlated with the severity of systemic inflammation. This study underscores the role of gut microbiota in driving liver injury mechanisms during TPN and supports a paradigm change in therapeutic targeting of the gut microbiota to mitigate TPN-related injury. KEY MESSAGES This study identified a differential link between gut microbiota and inflammation—the higher the dysbiosis, the worse the systemic inflammatory markers. Higher levels of Firmicutes species correlated with reduced inflammation.

Distinct Cecal and Fecal Microbiome Responses to Stress Are Accompanied by Sex- and Diet-Dependent Changes in Behavior and Gut Serotonin.

Although diet- and stress-induced perturbations in the microbiome (biotic and abiotic factors) associate with changes in host behavior via the microbiota-gut-brain axis, few mechanisms have been identified. The identification of causative pathways by which the microbiome influences host behavior therefore would benefit from the application of evidence-based conceptual frameworks. One such causal framework is microbial endocrinology which is the study of neuroendocrine axes as avenues of bi-directional neurochemical-based host-microbe crosstalk. As such, we investigated the relationship between diet- and stress-induced alterations in behavior, regional gut serotonergic response, and concomitant changes in the cecal and fecal bacterial populations of male and female mice. Our results demonstrate that sex is a dominant factor in determining compositional changes in the gut microbiome in response to stress and diet modifications. Intestinal serotonergic responses to stress were observed in both sexes but dietary modifications uniquely affected region-specific changes in males and females. Likewise, behavioral alterations diverged between male and female mice. Together, these results demonstrate distinct sex-dependent relationships between cecal and fecal bacterial taxa and behavioral- and serotonergic-responses to stress and diet. The present study demonstrates the importance of including both male and female sexes in the examination of the microbiota-gut-brain axis. As different microbial taxa were identified to associate with the behavioral and gut serotonergic responses of male and female mice, certain bacterial species may hold sex-dependent functional relevance for the host. Future investigations seeking to develop microbiome-based strategies to afford host stress resilience should include sex-based differences in the microbiome.

Seaweed Supplementation Failed to Affect Fecal Microbiota and Metabolome as Well as Fecal IgA and Apparent Nutrient Digestibility in Adult Dogs

Simple SummarySeaweeds represent a source of bioactive compounds that have recently drawn the attention of the scientific community for their possible application as health-promoting ingredients. In particular, their indigestible polysaccharides have exhibited promising prebiotic properties both in humans and in farm animals. The present study investigated for the first time in healthy adult dogs the effects of dietary supplementations with intact seaweeds (brown algae Ascophyllum nodosum, Undaria pinnatifida, Saccharina japonica and red alga Palmaria palmata) on some fecal bacterial populations and metabolites, fecal IgA and apparent total tract digestibility of nutrients. The different algal supplementations (fed to ten healthy adult dogs at a dietary dose of 15 g/kg for 28 days) did not have any significant effect on the selected fecal microbial parameters, intestinal immunity and nutrient digestibility. Further studies investigating higher dietary inclusions of intact seaweeds or their polysaccharide extracts are needed to gain a better understanding of the potential of these interesting marine resources in companion animal nutrition.The present study investigated in dogs the dietary effects of intact seaweeds on some fecal bacterial populations and metabolites, fecal IgA and apparent total tract digestibility (ATTD). Ten healthy adult dogs were enrolled in a 5 × 5 replicated Latin square design to evaluate five dietary treatments: control diet (CD); CD + Ascophyllum nodosum; CD + Undaria pinnatifida; CD + Saccharina japonica; CD + Palmaria palmata (n replicates per treatment = 10). Seaweeds were added to food at a daily dose of 15 g/kg. The CD contained silica as a digestion marker. Each feeding period lasted 28 d, with a 7 d wash-out in between. Feces were collected at days 21 and 28 of each period for chemical and microbiological analyses. Fecal samples were collected during the last five days of each period for ATTD assessment. Dogs showed good health conditions throughout the study. The fecal chemical parameters, fecal IgA and nutrient ATTD were not influenced by algal supplementation. Similarly, microbiological analyses did not reveal any effect by seaweed ingestion. In conclusion, algal supplementation at a dose of 15 g/kg of diet failed to exert noticeable effects on the canine fecal parameters evaluated in the present study.

Wheat Germ Supplementation Improves Glucose Homeostasis Markers of Overweight Adults Independent of Gut Microbial Modulation

Abstract Objectives This study investigated the effects of wheat germ (WG) supplementation on inflammation, metabolic, and gut health markers in overweight adults. Methods Forty overweight (BMI = 25.0–30 kg/m2) adults between 18–45 y old were recruited to this single-blinded randomized placebo-controlled study. Qualified participants were asked to maintain their normal diet and physical activity and consume energy balls containing 30 g of either WG or cornmeal (control) daily for 4 wks. Anthropometric and metabolic parameters, as well as dietary (3-d food record), medical history, physical activity (Yale Physical Activity Survey), stool measures (Bristol Stool Chart and the Cleveland Clinic Constipation Scoring System), gut integrity markers, and fecal bacterial population were assessed at baseline and at the end of the supplementation period. Results Thirty-nine participants completed the 4-week study (n = 20 and 19 for the WG and control group, respectively). There were no differences in the lipid profile, but glycated hemoglobin (P = 0.04), insulin (P = 0.03), and homeostatic model assessment of insulin resistance (HOMA-IR; P = 0.04) were significantly decreased in the WG but not the control group. Similarly, the adipokine resistin was also significantly reduced (P = 0.03) by WG but not the control. There were no changes in stool characteristics between the two groups before and after supplementation. Similarly, there were no changes in gut bacterial population due to WG supplementation but the phyla Bacteroidetes (P = 0.03) and Proteobacteria (P = 0.048) and the genus Bacteroides (P = 0.03) were significantly decreased in the control group. No significant changes were observed in plasma inflammatory markers, fecal short-chain fatty acid (SCFA) concentrations, and markers of gut integrity in both groups. Conclusions Four weeks of WG supplementation improved markers of glucose homeostasis, which can partly be attributed to the reduction of the pro-inflammatory adipokine resistin and not due to changes within the gut (i.e., bacterial population, gut integrity markers, and SCFA production). Our findings indicate that WG may be a safe, effective, and economical approach to improve glucose homeostasis. Funding Sources USDA Award #2019–67,018-29,260 and the Jim and Lynn Williams Professorship.

Dietary Fructooligosaccharides Reduce Mercury Levels in the Brain of Mice Exposed to Methylmercury.

Methylmercury (MeHg) exposure during pregnancy is a concern because of its potential health risks to fetuses. Intestinal microbiota has important roles in the decomposition and fecal excretion of MeHg. We investigated the effect of nondigestible saccharides on the accumulation and excretion of Hg after MeHg exposure. Female BALB/cByJ mice were fed a basal diet or the same diet supplemented with 5% fructooligosaccharides (FOS) or 2.5% glucomannan. Six weeks after feeding, mice were administered MeHg chloride (4 mg Hg/kg, per os (p.o.)), and urine and feces were collected for 28 d. FOS-fed mice had lower total Hg levels in all tissues (including the brain) compared with that of controls. The glucomannan diet had no effect on tissue Hg levels. No differences in tissue concentrations of inorganic Hg among groups were found. Fecal Hg excretion was markedly higher in FOS-fed mice than that in controls, but urinary Hg excretion was similar. FOS-fed mice had a higher proportion of inorganic Hg in feces than that of controls, with a significant increase in fecal Hg excretion. Analysis of fecal bacterial population showed the relative abundance of Bacteroides in FOS-fed mice to be higher than that in controls. The results suggest that FOS enhanced fecal Hg excretion and decreased tissue Hg levels after MeHg administration, possibly by accelerating MeHg demethylation by intestinal bacteria (the candidate genus Bacteroides). This demethylation also reduces MeHg absorption in the large intestine. In conclusion, daily FOS intake may decrease tissue Hg levels in animals and humans exposed to MeHg.

Evaluation the effect of subchronic feeding of transgenic cotton line (CKC1) on the faecal microbiota of albino rabbits

Recent studies have demonstrated a strong relationship between the intestinal microbiota and the host health. As such, consumers are increasingly becoming more concerned about the potential effect of certain foods/feeds, particularly of transgenic origin on the gut microbiota. Although the European Food Safety Authority has recommended in their guidelines, to study the effect of transgenic food/feed on host-microbiota, yet, few studies have focused on the evaluation of such effects mainly due to culturing difficulties. Therefore, this study was intended to evaluate the potential adverse effects of transgenic diet consumption on some specific gut microflora (Lactobacillus group, Bifidobacterium genus, Escherichia coli subgroup and Enterococcus genus) of rabbits. A total of forty-eight rabbits were randomly assigned into four groups and fed a diet containing a variable proportion of transgenic cottonseeds at 0, 20, 30 and 40% inclusion level, respectively. Changes in the specific or total faecal bacterial population were monitored at five different experimental stages (i.e. 0, 45, 90, 135 and 180days) using both the traditional plate count method (TM) and quantitative real-time PCR (qPCR). No significant differences (p>.05) were observed concerning numbers of specific bacteria or total bacteria between the control and experimental groups, though qPCR showed numerically higher values in terms of 16S rRNA gene copies as compared to the values obtained from TM. However, such numerical differences were biologically insignificant (p>.05). Similarly, no significant variations were noticed in the calculated B/E (log10 copies of Bifidobacterium per g faces/log10 copies of E. coli genome per g faeces) ratios in all the groups. All the ratios were in the range of 1.24 to 1.30 throughout the experiment, indicating a good balance of intestinal microflora and greater resistance to intestinal disorders. It is therefore concluded that feeding transgenic cottonseeds could not adversely affect the gut microflora of rabbits during a long-term study.

A Novel Corn-Expressed Phytase Improves Daily Weight Gain, Protein Efficiency Ratio and Nutrients Digestibility and Alters Fecal Microbiota in Pigs Fed with Very Low Protein Diets.

Simple SummaryFeeding pigs with very low crude protein (CP) diets with reduced calcium (Ca) and phosphorous (P) may be an effective strategy to decrease the nutrient excretion; however, this practice can negatively impact the growth performance of animals. Thus, there is an urgent need to improve the utilization of nitrogen, Ca and P in pigs. This study aimed to assess the effect of a novel corn-expressed phytase on growth performance, nutrients digestibility, gut microbial population, bone mineral density and content and blood metabolites in nursery pigs fed with low-CP, -Ca, and -P diets. Supplementation of very low protein diets with a corn-expressed phytase decreased the negative effects of these diets on average daily gain and protein efficiency ratio of pigs, increased the digestibility of Ca and P regardless of the levels of these minerals in the diet, improved bone characteristics and altered the fecal bacterial composition. This study suggests that corn-expressed phytase can be potentially useful for improving the growth performance of weaned pigs fed with low protein diets and should be considered for sustainable swine production. The objective of this study was to assess the effect of a novel corn-expressed phytase (CEP) on growth, nutrients digestibility, bone characteristics and fecal microbiota of pigs fed with very low-protein, -calcium (Ca) and -phosphorous (P) diets. Forty-eight barrows were subjected to 6 groups for 4 weeks: positive control-adequate protein (PC), negative control-reduced protein (NC), NC + low-dose CEP, i.e., 2000 FTU/kg (LD), NC + high-dose CEP, i.e., 4000 FTU/kg (HD), LD with 0.12% unit reduced Ca and 0.15% unit reduced available P (LDR), and HD with 0.12% unit reduced Ca and 0.15% unit reduced available P (HDR). Compared to NC, LD and HDR had a higher average daily gain (ADG) and gain:protein ratio (G:P), HD and HDR had greater apparent fecal digestibility of Ca and P and bone mineral density and LDR and HDR had lower serum osteocalcin. The feces of LD was enriched in Lachnospiraceae, while the HD had a higher abundance of Succinvibrio and LDR had a higher abundance of Bifidobacterium and Actinobacteria. In conclusion, supplementation of protein-restricted diets with a CEP decreased their negative effects on ADG and G:P ratio, increased the digestibility of Ca and P regardless of the levels of these minerals in the diet, improved bone characteristics and produced differential effects on fecal bacterial population.

Production of compost with biopesticide property from toxic weed Lantana: Quantification of alkaloids in compost and bacterial pathogen suppression

Toxic weed Lantana camara foliage was composted with cow dung in 2:1 and 1:1 ratio (v/v) and changes in physicochemical characteristics, and faecal coliform bacterial population (Escherichia coli and Salmonella) was estimated for 35 d. Results showed a significant increase in total N (1.48-1.69-folds), Paval (6.87–9.19-folds), and total K (1.08−1.23-folds) content, while a decrease in C/N ratio (1.87–2.13-folds) and total organic carbon (1.12−1.46-folds) after composting process. Germination index (GI) results (> 70 %) suggested the non-toxic property of Lantana compost against tested rapeseed mustard seeds. E. coli and Salmonella population reduced to the safe limit after 35 d composting. Compost extract (sterilized and non-sterilized) (from 2:1 setup) showed about 0.88 – 1.08-, 0.88 – 0.96-, 0.83 – 0.94-, and 0.79–1.08-folds higher inhibition in Xanthomonas citrus, Xanthomonas campestris, Erwinia carotovora, and Pseudomonas aerogenosa, respectively, indicating strong pathogen-inhibiting substances in Lantanacompost. GC–MS analysis of compost extract indicated the presence of isomers of several compounds of biocidal property – hexadecane (9-hexyl and 9-octyl); 2-tridecyl ester; eicosane; tetradecane, heptacosane (1-chloro- and 9-hexyl); heptadecane, octadecane, 3-ethyl-5-(2-ethylbutyl)-, heptacosane, tetradecane, 2,6,10-trimethyl-, etc.). Result revealed that Lantana compost could be used as biomanure with biopesticide properties for sustainable organic farming system.

Prevention of bone loss by using Lactobacillus-fermented milk products in a rat model of glucocorticoid-induced secondary osteoporosis

Secondary osteoporosis is a common side effect of glucocorticoid treatments. The anti-osteoporotic role of two fermented milk products (FMPs), produced with Lactobacillus plantarum A41 and Lactobacillus fermentum SRK414, in alleviating secondary osteoporotic symptoms induced by dexamethasone (Dex) injection in rats was investigated. The two FMPs could prevent Dex-induced bone loss in rats. The two FMPs normalised altered gene expression levels of bone metabolism-related markers (osteoprotegerin, osteocalcin, bone sialoprotein, receptor activator of nuclear factor kappa-B ligand, and tartrate-resistant acid phosphatase). Furthermore, they regulated gene expression levels of bone morphogenetic protein 2-related and cell apoptosis markers in the tibia and calcium metabolism-related markers in the ileum. Additionally, these treatments modulated faecal bacterial populations by increasing Lactobacillus colonies. FMPs fermented by L. plantarum A41 and L. fermentum SRK414 were capable of preventing glucocorticoid-induced secondary osteoporosis via regulation of intestinal microbiota and gene expression levels of bone cell apoptosis related markers.

Microbiome Signatures Associated with Rodent Model of Traumatic Brain Injury vs. Psychological Stress

Traumatic brain injury (TBI) and post‐traumatic stress disorder (PTSD) are two signature illnesses of modern warfare. Discrimination between psychological stress and TBI using a knowledge‐driven unbiased panel of biomarker signatures will be essential for designing precise care management. Recent data suggest a role of alterations of the gut micobiome architecture susceptible to both brain injury and psychological disorders. The present study was conducted to identify factors discriminating between psychological stress and TBI in the fecal microbiome of rats and further to study the effect of diets enriched with varied polyunsaturated fat compositions.A closed‐head TBI model consisting of blast overpressure (BOP) wave exposure coupled with a weight drop concussion (Marmarou method) was used on a group of adult male rats (N=12 each exposed and sham). In parallel, an independent group of rats (N=12 each exposed and sham) was subjected to an underwater trauma (UWT) stressor model that consisted of 30s of swimming and habituation, followed by 30s of forced whole body immersion. Prior to the stress types (BOP or UWT), animals were maintained for six weeks and continued thereafter on two different diets (N=4 per diet group): Standard house chow and custom chow. Unlike the house chow, the custom diets contained no long chain ω‐3 polyunsaturated fatty acids to offset their high Linoleic acid content. The fecal bacterial populations were characterized by identification of 16S ribosomal RNA.Principal coordinate analysis showed clear separation between taxonomic phylogenetic profiles linked to the stress types from their corresponding shams. Within each stress type, the rodents fed on house chow showed different fecal microbial population from the customized diet. Firmicutes, Proteobacteria, and Cyanobacteria were those phyla of interest that showed significant population variance caused by the stress and diet variants. We integrated this microbial profile with rodent blood and brain genomic profile assayed in parallel. The result suggested a significant contribution of gut‐microbiome in energy networks related to the stress response, and diet emerged as a strong candidate in regulating resilience. This outcome could lead in identifying novel customized treatment of TBI and psychological stress.Support or Funding InformationThe project was funded by DoD‐MOMRPThe differential impact of two neurological stress models on different component of gut‐brain axisFigure 1

Influence of sodium chlorate, ferulic acid, and essential oils on Escherichia coli and porcine fecal microbiota.

The influence of sodium chlorate (SC), ferulic acid (FA), and essential oils (EO) was examined on the survivability of two porcine diarrhetic enterotoxigenic Escherichia coli (ETEC) strains (F18 and K88) and populations of porcine fecal bacteria. Fecal bacterial populations were examined by denaturing gradient gel electrophoresis (DGGE) and identification by 16S gene sequencing. The treatments were control (no additives), 10 mM SC, 2.5 mg FA /mL, a 1.5% vol/vol solution of an EO mixture as well as mixtures of EO + SC, EO + FA, and FA + SC at each of the aforementioned concentrations. EO were a commercial blend of oregano oil and cinnamon oil with water and citric acid. Freshly collected porcine feces in half-strength Mueller Hinton broth was inoculated with E. coli F18 (Trial 1) or E. coli K88 (Trial 2). The fecal-E. coli suspensions were transferred to crimp top tubes preloaded with the treatment compounds. Quantitative enumeration was at 0, 6, and 24 h. All treatments reduced (P < 0.05) the counts of E. coli F18 at 6 and 24 h. With the exception of similarity coefficient (%SC), all the other treatments reduced (P < 0.05) the K88 counts at 24 h. The most effective treatments to reduce the F18 and K88 CFU numbers were those containing EO. Results of DGGE revealed that Dice percentage similarity coefficients (%SC) of bacterial profiles among treatment groups varied from 81.3% to 100%SC. The results of gene sequencing showed that, except for SC at 24 h, all the other treatments reduced the counts of the family Enterobacteriaceae, while Lactobacillaceae and Ruminococcaceae increased and Clostridiaceae decreased in all treatments. In conclusion, all treatments were effective in reducing the ETEC, but EO mixture was the most effective. The porcine microbial communities may be influenced by the studied treatments.

176 Characterizing the mycobiome in piglets during the weaning transition

Abstract The importance of the microbiota in the gastrointestinal (GI) tract of animals is recognized as a critical player in host health. Recently, the significance of the mycobiome has been recognized, but culture-independent studies are limited, especially in swine. Weaning is a time of stress, dietary changes, and a predisposition to infections, making it a time of interest to industry. In this study, we sought to assess and characterize the mycobiome and microbiome in the feces and GI tract of swine from birth through the critical weaning transition (days 1–35 post-birth). In addition, we investigated environmental factors that may alter microorganisms present in piglets. Fecal bacterial populations increased in diversity over the experimental timeline and demonstrated a transition from an Enterobacteriaceae-dominated population, to a Prevotellaceae and Ruminococcaceae-dominated population by days 24–35 post-birth. These later populations are capable of fiber degradation and short chain fatty acid production. In fecal fungal populations, richness and diversity peaked at weaning and declined post-weaning. There was also a dynamic shift in the mycobiome to a Saccharomycetaceae-dominated population that remained stable into adulthood. Fungal organisms contributing to this colonization were not found in environmental samples including water, colostrum, and feed. Despite fungal populations present in the feces of sows, these maternal fungi were not similar to the piglet mycobiome and thus did not indicate a maternally-derived effect. Furthermore, the microbiomes of the GI tract showed decreased richness and diversity in the upper GI compared to the lower GI, and a high degree of individual variation and litter effect throughout the organs. This study provides insights into the early colonization and establishment of fungi during the weaning transition. Future studies will investigate the effect of the mycobiome on piglet growth and health during the weaning transition, including their role in fast- versus slow-growing piglets.

Production Performance, Egg Quality, and Fecal Bacterial Population of Laying Ducks Fed Ration Supplemented by Bamboo Vinegar

With the ban on the use of antibiotics in poultry feed, it is required to produce alternative substances to maintain poultry health and performance. The present study was carried out to investigate the effects of dietary supplementation at various levels of bamboo vinegar (BV) on performance, egg quality, and fecal bacterial population of laying ducks. One hundred and five 50-week-old crossbred laying ducks (Khaki Campbell × Native) were randomly allocated into five treatment groups, each with seven replicates of three ducks. The experimental ducks were fed a basal diet (antibiotic-free diet) supplemented with BV at the levels of 0%, 0.2%, 0.4%, 0.6%, or 0.8% of diet until 62 weeks of age. For the overall period (from 50 to 62 weeks of age), the dietary supplementation of BV at the levels of 0.4% and 0.6% of diet increased egg production and egg mass, and improved feed efficiency of laying ducks (p 0.05). The population of fecal Escherichia coli was significantly decreased in the experimental ducks fed diet supplemented with BV at the levels of 0.4%, 0.6%, and 0.8% compared to the control group (p<0.01), while the population of Lactobacillus spp. remained unchanged. The present results indicate that a level of 0.4% BV supplementation in laying duck diet is sufficient for reducing fecal E. coli and improves productive performance of egg laying ducks.

Anti-Atherosclerotic Properties of Wild Rice in Low-Density Lipoprotein Receptor Knockout Mice: The Gut Microbiome, Cytokines, and Metabolomics Study.

Background and aim: We previously reported the anti-atherogenic properties of wild rice in low-density lipoprotein receptor knockout (LDL-r-KO) mice. The present study aimed to discover the mechanism of action for such effects. Materials: Fecal and plasma samples from the wild rice treated and control mice were used. Fecal bacterial population was estimated while using 16S rDNA technology. The plasma samples were used to estimate the levels of 35 inflammatory markers and metabolomics, while using Meso Scale multiplex assay and liquid chromatography-mass spectrometry (LC-MS/MS) techniques. Results: Many bacteria, particularly Anaeroplasma sp., Acetatifactor sp., and Prophyromonadaceae sp., were found in higher quantities in the feces of wild rice fed mice as compared to the controls. Cytokine profiles were significantly different between the plasma of treated and control mice. Among them, an increase in the level of IL-10 and erythropoietin (EPO) could explain the anti-atherogenic properties of wild rice. Among many metabolites tested in plasma of these animals, surprisingly, we found an approximately 60% increase in the levels of glucose in the wild rice fed mice as compared to that in the control mice. Conclusion: Additional studies warrant further investigation of the interplay among gut microbiome, inflammatory status, and macronutrient metabolism.

Fecal characteristics and gut bacterial population of weaned pigs fed amino acid-supplemented diets varying in crude protein and fermentable carbohydrate contents

A 28-day study was conducted to evaluate the effects of dietary crude protein (CP; high- and low-CP diets) and dried citrus pulp (DCP; 0 and 7.5%) contents on fecal consistency score and intestinal bacterial enumeration. A total of 108 early-weaned piglets (21 days of age; 5.82 ± 0.16 kg initial body weight) were assigned to 1 of 4 dietary treatments in a 2 × 2 factorial arrangement, with 9 replicate pens per treatment and 3 pigs per pen. The high-CP diets consisted of feeding 20 and 21% CP contents throughout phase 1 (1 to 14 day) and phase 2 (15 to 28 day), respectively. For the low-CP diets, CP contents were reduced by 4 percentage units as compared with the high-CP diets in both phases. Crystalline amino acids (AA) were supplemented to maintain an ideal AA pattern. Feces were scored for consistency during the first 7 days postweaning. Digesta samples from the ileum, cecum, and colon were collected on days 7 and 28 postweaning to determine total coliform and lactobacilli counts. There were no CP x DCP interactions for fecal consistency scores. Although fecal consistency scores were unaffected by the dietary CP contents, adding 7.5% DCP to the diet reduced (P ≤ 0.03; days 1 and 3) and tended to reduce (P ≤ 0.09; days 2 and 5) the incidence of soft feces. On day 7 postweaning, reducing dietary CP inclusion resulted in decreased (P < 0.05) ileal and colonic coliform population in pigs fed diets containing DCP. Additionally, adding 7.5% DCP to the diet decreased (P < 0.05) cecal coliform counts only for pigs fed the low-CP diet on day 7. On day 28 postweaning, there were no treatment effects on coliform population in the ileum, cecum, and colon. Ileal, cecal, and colonic counts of lactobacilli were similar among treatments on days 7 and 28 postweaning. An effect of sampling day was found for ileal coliform (P < 0.01) and cecal lactobacilli (P = 0.04) counts, with greater bacterial populations observed on day 7 when compared with day 28 postweaning. In summary, adding 7.5% DCP to low-CP AA-supplemented diets decreased coliform bacteria proliferation in piglet hindgut immediately after weaning.