Gastrointestinal Bacterial Community Research Articles

Identification of Gastrointestinal Microbiota in Hawaiian Green Turtles (Chelonia mydas).

Green turtles (Chelonia mydas) have a hindgut fermentation digestive tract, which uses cellulolytic microbes to break down plant matter in the cecum and proximal colon. Previous studies on bacterial communities of green turtles have not identified in situ hindgut microbiota, and never before in Hawaiian green turtles, which comprise an isolated metapopulation. Fresh samples using sterile swabs were taken from five locations along the gastrointestinal tracts of eight green turtles that had required euthanization. Bacteria were cultured, aerobically and anaerobically, on nutrient agar and four differential and selective media. Samples at three sections along the gastrointestinal tracts of two green turtles were analyzed using 16S metagenomics on an Ion Torrent Personal Genome Machine. More than half of the 4 532 104 sequences belonged to the phylum Firmicutes, followed by Bacteroidetes and Proteobacteria, which are characteristic of herbivore gut microbiota. Some microbiota variation existed between turtles and among gastrointestinal sections. The 16S sequence analysis provided a better representation of the total gastrointestinal bacterial community, much of which cannot be cultured using traditional microbial techniques. These metagenomic analyses serve as a foundation for a better understanding of the microbiome of green turtles in the Hawaiian Islands and elsewhere.

Altered Stool Microbiota of Infants with Cystic Fibrosis Shows a Reduction in Genera Associated with Immune Programming from Birth

Previous work from our group indicated an association between the gastrointestinal microbiota of infants with cystic fibrosis (CF) and airway disease in this population. Here we report that stool microbiota of infants with CF demonstrates an altered but largely unchanging within-individual bacterial diversity (alpha diversity) over the first year of life, in contrast to the infants without CF (control cohort), which showed the expected increase in alpha diversity over the first year. The beta diversity, or between-sample diversity, of these two cohorts was significantly different over the first year of life and was statistically significantly associated with airway exacerbations, confirming our earlier findings. Compared with control infants, infants with CF had reduced levels of Bacteroides, a bacterial genus associated with immune modulation, as early as 6 weeks of life, and this significant reduction of Bacteroides spp. in the cohort with CF persisted over the entire first year of life. Only two other genera were significantly different across the first year of life: Roseburia was significantly reduced and Veillonella was significantly increased. Other genera showed differences between the two cohorts but only at selected time points. In vitro studies demonstrated that exposure of the apical face of polarized intestinal cell lines to Bacteroides species supernatants significantly reduced production of interleukin 8 (IL-8), suggesting a mechanism whereby changes in the intestinal microbiota could impact inflammation in CF. This work further establishes an association between gastrointestinal microbiota, inflammation, and airway disease in infants with CF and presents a potential opportunity for therapeutic interventions beginning in early life.IMPORTANCE There is growing evidence for a link between gastrointestinal bacterial communities and airway disease progression in CF. We demonstrate that infants with CF ≤1 year of age show a distinct stool microbiota versus that of control infants of a comparable age. We detected associations between the gut microbiome and airway exacerbation events in the cohort of infants with CF, and in vitro studies provided one possible mechanism for this observation. These data clarify that current therapeutics do not establish in infants with CF a gastrointestinal microbiota like that in healthy infants, and we suggest that interventions that direct the gastrointestinal microbiota closer to a healthy state may provide systemic benefits to these patients during a critical window of immune programming that might have implications for lifelong health.

Characterization of the gastrointestinal bacterial microbiome of farmed juvenile and adult white Cachama (Piaractus brachypomus)

The bacterial communities associated to the gastrointestinal tract in fish, are involved in physiology, development, and immune responses against pathogenic bacteria. However, for Piaractus brachypomus (White Cachama) a tropical fresh water fish from the Amazonas and Orinoco basins and the second most produced fish in Colombia, these communities are still largely unknown. The study of bacterial communities and how they are affected have increased in fish with economic value. In this study, reared individuals of P. brachypomus were chosen to investigate the structure of intestinal microbiota by culture-dependent methods, Denaturing Gradient Gel Electrophoresis (DGGE) and 16S rRNA gene survey, taking into consideration the anterior and posterior gastrointestinal tract of juvenile and adult fish. Results showed that intestinal microbiota of P. brachypomus was dominated by Fusobacteria, Spirochaetes, Firmicutes and Proteobacteria across life stages and gastrointestinal sites. We observed statistically significant differences in beta-diversity between life stages. Some genera significantly higher in juvenile individuals e.g. Brevinema, had a very low abundance in adult ones, suggesting that developmental stage may select some bacterial groups, and Cetobacterium, a B12 vitamin producing bacteria, showed significant dominance across gut sites and life stages, suggesting an important role in P. brachypomus digestion. Culture dependent techniques allowed the isolation of bacteria of interest in aquaculture, previously reported fish probiotic and pathogenic bacteria were obtained. The combination of different approaches allowed the first assessment of the gastrointestinal associated bacterial communities of P. brachypomus, showing that developmental has an influence on its composition and structure.

Changes in Cold and Hot Syndrome and Gastrointestinal Bacterial Community Structure in Mice by Intervention with Food of Different Nature.

To reveal the effect of foods with different natures on cold or hot syndrome and gastrointestinal bacterial community structure in mice. Forty-five 6-week-old male ICR Kunming mice of clean grade were divided into 5 groups, 9 per group, including the control (CK), hot nature herbs (HM), Hong Qu glutinous rice wine (RW), tea rice wine (TW), and cold nature herbs (CM) groups. Distilled water or corresponding herbs were administered to mice (0.01 mL/g body weight) in the 5 groups by gastric infusion respectively, once daily for 28 d. Appearance, behavior, and serum biochemical indicators, including 5-hydroxytryptamine (5-HT), thyroid stimulating hormone (TSH), noradrenaline (NE), cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), the hot nature index, as well as the gastrointestinal bacterial community structure were analyzed in all groups after treatment. After supplementation for 28 d, CM and TW mice showed different degrees of cold syndrome, and HM and RW mice showed different degrees of hot syndrome. Compared with the HM and RW mice, the TSH, NE, cAMP levels and hot nature indices in the CM and TW mice were significantly decreased and 5-HT and cGMP levels were significantly increased (P<0.05). There was no obvious change in appearance or behavior in CK mice. Results of clustering analysis showed that the gastrointestinal bacterial community structures were highly similar in TW and CM mice as well as in RW and HM mice, and that they were from the same branch, respectively, when the distance was 0.02. The key microbes associated with cold syndrome were Lachnospiraceae uncultured, Lactococcus, etc., and the key microbes associated with hot syndrome were S24-7 norank, Ruminococcaceae uncultured, etc. CONCLUSION: The interventions with different nature foods could change cold or hot syndrome in mice, leading to changes in gastrointestinal bacterial community structure.

Strong Multivariate Relations Exist Among Milk, Oral, and Fecal Microbiomes in Mother-Infant Dyads During the First Six Months Postpartum

ABSTRACTBackgroundNeonatal gastrointestinal (GI) bacterial community structure may be related to bacterial communities of the mother, including those of her milk. However, very little is known about the diversity in and relationships among complex bacterial communities in mother-infant dyads.ObjectiveOur primary objective was to assess whether microbiomes of milk are associated with those of oral and fecal samples of healthy lactating women and their infants.MethodsSamples were collected 9 times from day 2 to 6 mo postpartum from 21 healthy lactating women and their infants. Milk was collected via complete breast expression, oral samples via swabs, and fecal samples from tissue (mothers) and diapers (infants). Microbiomes were characterized using high-throughput sequencing of the 16S ribosomal RNA (rRNA) gene. Alpha and beta diversity indices were used to compare microbiomes across time and sample types. Membership and composition of microbiomes were analyzed using nonmetric multidimensional scaling and canonical correlation analysis (CCA). The contribution of various bacterial communities of the mother-infant dyad to both milk and infant fecal bacterial communities were estimated using SourceTracker2.ResultsBacterial community structures were relatively unique to each sample type. The most abundant genus in milk and maternal and infant oral samples was Streptococcus (47.1% ± 2.3%, 53.9% ± 1.3%, and 69.1% ± 1.8%, respectively), whereas Bacteroides were predominant in maternal and infant fecal microbiomes (22.9% ± 1.3% and 21.4% ± 2.4%, respectively). The milk microbiome was more similar to the infant oral microbiome than the infant fecal microbiome. However, CCA suggested strong associations between the complex microbial communities of milk and those of all other sample types collected.ConclusionsThese findings suggest complex microbial interactions between breastfeeding mothers and their infants and support the hypothesis that variation in the milk microbiome may influence the infant GI microbiome.

Bacterial community structure and function distinguish gut sites in captive red-shanked doucs (Pygathrix nemaeus).

The mammalian order primates contains wide species diversity. Members of the subfamily Colobinae are unique amongst extant primates in that their gastrointestinal systems more closely resemble those of ruminants than other members of the primate order. In the growing literature surrounding nonhuman primate microbiomes, analysis of microbial communities has been limited to the hindgut, since few studies have captured data on other gut sites, including the foregut of colobine primates. In this study, we used the red-shanked douc (Pygathrix nemaeus) as a model for colobine primates to study the relationship between gastrointestinal bacterial community structure and gut site within and between subjects. We analyzed fecal and pregastric stomach content samples, representative of the hindgut and foregut respectively, using 16S recombinant DNA (rDNA) sequencing and identified microbiota using closed-reference operational taxonomic unit (OTU) picking against the GreenGenes database. Our results show divergent bacterial communities clearly distinguish the foregut and hindgut microbiomes. We found higher bacterial biodiversity and a higher Firmicutes:Bacteroides ratio in the hindgut as opposed to the foregut. These gut sites showed strong associations with bacterial function. Specifically, energy metabolism was upregulated in the hindgut, whereas detoxification was increased in the foregut. Our results suggest a red-shanked douc's foregut microbiome is no more concordant with its own hindgut than it is with any other red-shanked douc's hindgut microbiome, thus reinforcing the notion that the bacterial communities of the foregut and hindgut are distinctly unique. OPEN PRACTICES: This article has been awarded Open Materials and Open Data badges. All materials and data are publicly accessible via the IRIS Repository at https://www.iris-database.org/iris/app/home/detail?id=york:934328. Learn more about the Open Practices badges from the Center for Open Science: https://osf.io/tvyxz/wiki.

Bacterial community and metabolome shifts in the cecum and colon of captive sika deer (Cervus nippon) from birth to post weaning.

Ruminant productivity is associated with the gastrointestinal tract bacterial community, which matures progressively with animal growth. However, knowledge of the bacteriome and metabolome dynamics in cecum and colon during the early lives of neonatal ruminants is limited. Thus, we examined the bacteriome and metabolomes of the cecum and colon in neonatal sika deer at days 1, 42 and 70. The bacterial diversity and richness in the cecum and colon increased with age, and the bacterial community significantly changed across three time points. For cecum and colon, the proportions of Bacteroides spp., Escherichia-Shigella, Clostridium sensu stricto 1, Lachnoclostridium spp. and Lactobacillus spp. were predominated at day 1 and decreased with age, while the proportions of Ruminococcaceae UCG 005, Ruminococcaceae UCG 010, Rikenellaceae RC9 and Prevotellaceae UCG 003 were predominated at days 42 and 70 and increased with age. The concentrations of creatine and serine were significantly decreased, whereas the concentrations of total short-chain volatile fatty acids, pelargonic acid and leucine were increased in both the cecum and colon during development. These data document the development of the bacterial community and the metabolites in the cecum and colon of sika deer, and suggest a possible importance of ecology niche on gut development.

614. Modeling Changes in Gastrointestinal and Respiratory Tract Bacterial Community Diversity Attributable to Common Antibiotic Exposures During Long-Term Acute Care

BackgroundReduced gastrointestinal tract bacterial community diversity has been associated with increased risk for healthcare-associated infections, including Clostridium difficile infection. We sought to develop a model for concomitant change in bacterial community diversity at gastrointestinal and respiratory tract sites, drawing upon a recently completed cohort study of 92 subjects recruited from a long-term acute care hospital (LTACH) for dense longitudinal oral, endotracheal aspirate (ET), and stool specimen collection.MethodsWe evaluated the first 30 subjects enrolled from the LTACH cohort, for whom complete antibiotic administration data and 16S rRNA gene (V1–V2 amplicon) sequencing data were available. Sequencing was performed via the Illumina HiSeq platform; operational taxonomic units (OTUs) were formed and taxonomy assigned (GreenGenes 13.8) via the QIIME 1.9.1 pipeline. Generalized linear mixed effects models were fit using R (3.5.0), Stan (2.1.7), via the “rstan” and “rethinking” packages.ResultsWe evaluated 472 subject-days of study enrollment across the 30 subjects (median 15 days/subject). ET specimens were available for all subject-days; oral and stool for 357 and 177 subject-days, respectively. We modeled daily change in Shannon diversity across oral, ET, and stool specimens, parameterized with daily exposure to cefepime, piperacillin–tazobactam, meropenem, IV vancomycin, and oral vancomycin. All parameters fit with Rhat value lower than 1.1. Absent antibiotic exposure on the previous day, the daily change in Shannon diversity at all sites was near zero. The largest observed effect was oral vancomycin on stool (daily delta Shannon: −0.6, 95% CI: −1.38 to 0.09). All estimated effects for intravenous antibiotics on the stool, and for all antibiotics at other sites were smaller.ConclusionSmall daily changes in bacterial community diversity were attributable to individual antibiotics, but all 95% certainty intervals crossed zero in this pilot study. Further work will focus on modeling specific taxonomic changes attributable to individual antibiotics and antibiotic interactions.Disclosures All authors: No reported disclosures.

Fecal bacterial diversity of wild Sichuan snub-nosed monkeys (Rhinopithecus roxellana).

The gastrointestinal tract of primates harbors a complex microbial community, playing an essential role in the degradation of otherwise indigestible structural carbohydrates. The phylogenetic and functional diversity of the bacterial community in the feces as a surrogate for the gastrointestinal tract of wild Sichuan snub-nosed monkeys (Rhinopithecus roxellana, N = 6) was characterized based on sequence analysis of 16S rRNA genes. A sex comparison was conducted, with a prior hypothesis that the abundances of the bacterial taxa and/or functional categories associated with energy and nutrient metabolism would be higher in adult females (N = 3) due to the higher reproductive costs compared to adult males (N = 3). Ten phyla were identified in all samples, among which Bacteroidetes and Firmicutes were the predominant. Included in the above two phyla, the members of Prevotellaceae (Prevotella in particular) and Ruminococcaceae were highly abundant, which are common bacteria in the gastrointestinal tract of primates and can degrade various structural carbohydrates such as cellulose, hemicellulose, and pectin. This functionality was in line with the high abundances of the metagenomes associated with carbohydrate metabolism. Consistent with our hypothesis, the abundances of the metagenomes associated with energy metabolism, folding/sorting and degradation, glycan biosynthesis and metabolism, and metabolism of amino acids were higher in adult females relative to adult males. Sex differences were also detected in the bacterial community structure, although no sex differences in the proportions of any bacterial taxa were found likely due to the small sample size. These results suggested that gastrointestinal bacterial communities may aid adult females in increasing energy and nutrition utilization efficiencies compared to adult males. Fecal bacterial communities were found to be more similar between individuals in adult females than in adult males. Our study presented the first examination of the fecal bacterial diversity of a little-studied, endangered foregut fermenter.

Analysing food-derived interactions between tourists and sika deer (Cervus nippon) at Miyajima Island in Hiroshima, Japan: implications for the physical health of deer in an anthropogenic environment

ABSTRACTSika deer (Cervus nippon) on Miyajima Island in Hiroshima, Japan, are accustomed to a human environment and close tourist-deer interactions occur. In 2008, city officials banned deer feeding in response to an increasing number of human-deer conflicts. Nevertheless, this regulation remains ineffective. In this study, food-derived interactions between tourists and deer were analyzed, and a faecal analysis was conducted to examine the effects of potentially beneficial bacteria on the digestive system of deer. Over 64 hours of observation, a total of 397 tourist-deer interactions were recorded. Most interactions involved tourists’ food purchases from street stalls (49.6%). The initiator of each interaction was recorded for 267 events (67.3%), and it was found that deer initiated nearly twice as many interactions as tourists (tourists: 93 events; deer: 174 events). However, feeding occurred in only 11.5% of deer-initiated interactions, while feeding occurred in 50.5% of tourist-initiated interactions. The analysis of gastrointestinal bacterial community compositions showed that deer in the tourism district possessed a lower portion of the order Lactobacellales than deer in the non-tourism district. This was presumably due to different food sources, indicating that the human-influenced environment, of which feeding is one element, could affect the physical health of the deer.

Comparison of the gut microbiota composition between wild and captive sika deer (Cervus nippon hortulorum) from feces by high-throughput sequencing

The gut microbiota is characterized as a complex ecosystem that has effects on health and diseases of host with the interactions of many other factors together. Sika deer is the national level for the protection of wild animals in China. The available sequencing data of gut microbiota from feces of wild sika deer, especially for Cervus nippon hortulorum in Northeast China, are limited. Here, we characterized the gastrointestinal bacterial communities of wild (7 samples) and captive (12 samples) sika deer from feces, and compared their gut microbiota by analyzing the V3–V4 region of 16S rRNA gene using high-throughput sequencing technology on the Illumina Hiseq platform. Firmicutes (77.624%), Bacteroidetes (18.288%) and Tenericutes (1.342%) were the most predominant phyla in wild sika deer. While in captive sika deer, Firmicutes (50.710%) was the dominant phylum, followed by Bacteroidetes (31.996%) and Proteobacteria (4.806%). A total of 9 major phyla, 22 families and 30 genera among gastrointestinal bacterial communities showed significant differences between wild and captive sika deer. The specific function and mechanism of Tenericutes in wild sika deer need further study. Our results indicated that captive sika deer in farm had higher fecal bacterial diversity than the wild. Abundance and quantity of diet source for sika deer played crucial role in shaping the composition and structure of gut microbiota.

Antibiotic treatment disrupts bacterial communities in the colon and rectum of simian immunodeficiency virus-infected macaques.

Antibiotic therapies are known to disrupt gastrointestinal (GI) bacterial communities. HIV and pathogenic simian immunodeficiency virus (SIV) infections have also been associated with disrupted GI bacterial communities. We administered a combination antibiotic therapy to six SIV-infected rhesus macaques and collected colon biopsies, stool samples and rectal swabs before and after antibiotics, and evaluated the bacterial communities at each sample site using high-throughput 16S rRNA gene sequencing. The colon mucosa and stool samples displayed different bacterial communities, while the rectal swabs showed a mixture of the mucosal and stool-associated bacteria. Antibiotics disrupted the native bacterial communities at each sample site. The colon mucosa showed depleted abundances of the dominant Helicobacteraceae, while we found depleted abundances of the dominant Ruminococcaceae sp. in the stool. The rectal swabs showed similar trends as the colon mucosa, but were more variable. After the antibiotic treatment, there were increased abundances of similar taxa of facultative anaerobic bacteria, including Lactobacillaceae and Enterobacteriaceae at each sample site.

Evolution of the digestive enzymes and bacterial changes of the gastrointestinal tract of the Artemia urmiana during growth period

As the digestive enzymes and gastro-intestinal (GI) bacterial community contribute to the health and nutrition of the organism, this study aims to evaluate the ontogeny of bacterial population and digestive enzymes activities in the GI tract of Artemia urmiana from nauplii to the adult stage. Artemia cysts were hatched under standard conditions and stocked at a density of 20 nauplii ml-1 for 15 days. Samplings for growth, bacterial and enzymatic analysis were collected on days 1, 5, 10 and 15 of the experiment. The results indicated that the GI tract was sterile at the time of hatching. Artemia GI tract was active after hatching by protease (0.282 ± 0.001 U mg-1 protein min-1), lipase (0.182 ± 0.001 U mg-1 protein min-1) and amylase (0.295 ± 0.001 U mg-1 protein min-1) secretion and increased during the experiment. In addition, a significant relation (p < 0.05) was observed between Artemia total length and the activity of digestive enzymes (lipase (r2 = 0.98) amylase (r2 = 0.97) and protease (r2 = 0.98). A significant relation (p < 0.05) was observed between total aerobic bacteria and digestive enzyme secretion (lipase, r2 = 0.83; amylase, r2 = 0.66 and protease, r2 = 0.84) too. Such a relation was observed between total Bacillus spp. count and digestive enzyme activities (lipase r2 = 0.82; amylase r2 = 0.79; and protease; r2 = 0.74). These results suggest that in addition to the chemical composition of food, total length, GI bacteria enzyme secretion and the interaction of these factors contribute in digestive enzymes ontogeny during the growth period.

Influence of dairy by-product waste milk on the microbiomes of different gastrointestinal tract components in pre-weaned dairy calves

The community structure of colonised bacteria in the gastrointestinal tracts (GITs) of pre-weaned calves is affected by extrinsic factors, such as the genetics and diet of the calves; however, the dietary impact is not fully understood and warrants further research. Our study revealed that a total of 6, 5, 2 and 10 bacterial genera showed biologically significant differences in the GITs of pre-weaned calves fed four waste-milk diets: acidified waste milk, pasteurised waste milk, untreated bulk milk, and untreated waste milk, respectively. Specifically, generic biomarkers were observed in the rumen (e.g., Bifidobacterium, Parabacteroides, Fibrobacter, Clostridium, etc.), caecum (e.g., Faecalibacterium, Oxalobacter, Odoribacter, etc.) and colon (e.g., Megamonas, Comamonas, Stenotrophomonas, etc.) but not in the faeces. In addition, the predicted metabolic pathways showed that the expression of genes related to metabolic diseases was increased in the calves fed untreated waste milk, which indicated that untreated waste milk is not a suitable liquid diet for pre-weaned calves. This is the first study to demonstrate how different types of waste milk fed to pre-weaned calves affect the community structure of colonised bacteria, and the results may provide insights for the intentional adjustment of diets and gastrointestinal bacterial communities.

Faecal microbiota transplantation protects against radiation‐induced toxicity

Severe radiation exposure may cause acute radiation syndrome, a possibly fatal condition requiring effective therapy. Gut microbiota can be manipulated to fight against many diseases. We explored whether intestinal microbe transplantation could alleviate radiation‐induced toxicity. High‐throughput sequencing showed that gastrointestinal bacterial community composition differed between male and female mice and was associated with susceptibility to radiation toxicity. Faecal microbiota transplantation (FMT) increased the survival rate of irradiated animals, elevated peripheral white blood cell counts and improved gastrointestinal tract function and intestinal epithelial integrity in irradiated male and female mice. FMT preserved the intestinal bacterial composition and retained mRNA and long non‐coding RNA expression profiles of host small intestines in a sex‐specific fashion. Despite promoting angiogenesis, sex‐matched FMT did not accelerate the proliferation of cancer cells in vivo. FMT might serve as a therapeutic to mitigate radiation‐induced toxicity and improve the prognosis of tumour patients after radiotherapy.

Metabolic Effects of Berries with Structurally Diverse Anthocyanins.

Overconsumption of energy dense foods and sedentary lifestyle are considered as major causes of obesity-associated insulin resistance and abnormal glucose metabolism. Results from both cohort studies and randomized trials suggested that anthocyanins from berries may lower metabolic risks, however these reports are equivocal. The present study was designed to examine effects of six berries with structurally diverse anthocyanin profiles (normalized to 400 µg/g total anthocyanin content) on development of metabolic risk factors in the C57BL/6 mouse model of polygenic obesity. Diets supplemented with blackberry (mono-glycosylated cyanidins), black raspberry (acylated mono-glycosylated cyanidins), blackcurrant (mono- and di-glycosylated cyanidins and delphinidins), maqui berry (di-glycosylated delphinidins), Concord grape (acylated mono-glycosylated delphinidins and petunidins), and blueberry (mono-glycosylated delphinidins, malvidins, and petunidins) showed a prominent discrepancy between biological activities of delphinidin/malvidin-versus cyanidin-type anthocyanins that could be explained by differences in their structure and metabolism in the gut. Consumption of berries also resulted in a strong shift in the gastrointestinal bacterial communities towards obligate anaerobes that correlated with decrease in the gastrointestinal luminal oxygen and oxidative stress. Further work is needed to understand mechanisms that lead to nearly anoxic conditions in the gut lumens, including the relative contributions of host, diet and/or microbial oxidative activity, and their implication to human health.

Characterization of the microbial communities along the gastrointestinal tract of sheep by 454 pyrosequencing analysis.

ObjectiveThe gastrointestinal tract of sheep contain complex microbial communities that influence numerous aspects of the sheep’s health and development. The objective of this study was to analyze the composition and diversity of the microbiota in the gastrointestinal tract sections (rumen, reticulum, omasum, abomasum, duodenum, jejunum, ileum, cecum, colon, and rectum) of sheep.MethodsThis analysis was performed by 454 pyrosequencing using the V3-V6 region of the 16S rRNA genes. Samples were collected from five healthy, small tailed Han sheep aged 10 months, obtained at market. The bacterial composition of sheep gastrointestinal microbiota was investigated at the phylum, class, order, family, genus, and species levels.ResultsThe dominant bacterial phyla in the entire gastrointestinal sections were Firmicutes, Bacteroidetes, and Proteobacteria. In the stomach, the three most dominant genera in the sheep were Prevotella, unclassified Lachnospiraceae, and Butyrivibrio. In the small intestine, the three most dominant genera in the sheep were Escherichia, unclassified Lachnospiraceae, and Ruminococcus. In the large intestine, the three most dominant genera in the sheep were Ruminococcus, unclassified Ruminococcaceae, and Prevotella. R. flavefaciens, B. fibrisolvens, and S. ruminantium were three most dominant species in the sheep gastrointestinal tract. Principal Coordinates Analysis showed that the microbial communities from each gastrointestinal section could be separated into three groups according to similarity of community composition: stomach (rumen, reticulum, omasum, and abomasum), small intestine (duodenum, jejunum, and ileum), and large intestine (cecum, colon, and rectum).ConclusionThis is the first study to characterize the entire gastrointestinal microbiota in sheep by use of 16S rRNA gene amplicon pyrosequencing, expanding our knowledge of the gastrointestinal bacterial community of sheep.

SIV Infection-Mediated Changes in Gastrointestinal Bacterial Microbiome and Virome Are Associated with Immunodeficiency and Prevented by Vaccination.

AIDS caused by simian immunodeficiency virus (SIV) infection is associated with gastrointestinal disease, systemic immune activation, and, in cross-sectional studies, changes in the enteric virome. Here we performed a longitudinal study of a vaccine cohort to define the natural history of changes in the fecal metagenome in SIV-infected monkeys. Matched rhesus macaques were either uninfected or intrarectally challenged with SIV, with a subset receiving the Ad26 vaccine, an adenovirus vector expressing the viral Env/Gag/Pol antigens. Progression of SIV infection to AIDS was associated with increased detection of potentially pathogenic viruses and bacterial enteropathogens. Specifically, adenoviruses were associated with an increased incidence of gastrointestinal disease and AIDS-related mortality. Viral and bacterial enteropathogens were largely absent from animals protected by the vaccine. These data suggest that the SIV-associated gastrointestinal disease is associated with the presence of both viral and bacterial enteropathogens and that protection against SIV infection by vaccination prevents enteropathogen emergence.

Spatial and Temporal Changes in the Broiler Chicken Cecal and Fecal Microbiomes and Correlations of Bacterial Taxa with Cytokine Gene Expression.

To better understand the ecology of the poultry gastrointestinal (GI) microbiome and its interactions with the host, we compared GI bacterial communities by sample type (fecal or cecal), time (1, 3, and 6 weeks posthatch), and experimental pen (1, 2, 3, or 4), and measured cecal mRNA transcription of the cytokines IL18, IL1β, and IL6, IL10, and TGF-β4. The microbiome was characterized by sequencing of 16S rRNA gene amplicons, and cytokine gene expression was measured by a panel of quantitative-PCR assays targeting mRNAs. Significant differences were observed in the microbiome by GI location (fecal versus cecal) and bird age as determined by permutational MANOVA and UniFrac phylogenetic hypothesis tests. At 1-week posthatch, bacterial genera significantly over-represented in fecal versus cecal samples included Gallibacterium and Lactobacillus, while the genus Bacteroides was significantly more abundant in the cecum. By 6-week posthatch, Clostridium and Caloramator (also a Clostridiales) sequence types had increased significantly in the cecum and Lactobacillus remained over-represented in fecal samples. In the ceca, the relative abundance of sequences classified as Clostridium increased by ca. 10-fold each sampling period from 0.1% at 1 week to 1% at 3 week and 18% at 6 week. Increasing community complexity through time were observed in increased taxonomic richness and diversity. IL18 and IL1β significantly (p < 0.05, pairwise t-tests) increased to maximum mean expression levels 1.5 fold greater at week 3 than 1, while IL6 significantly decreased to 0.8- and 0.5-fold expression at 3- and 6-week posthatch, respectively relative to week 1. Transcription of pro-inflammatory cytokines was generally negatively correlated with the relative abundance of various members of the phylum Firmicutes and positively correlated with Proteobacteria. Correlations of the microbiome with specific cytokine mRNA transcription highlight the importance of the GI microbiome for bird health and productivity and may be a successful high-throughput strategy to identify bacterial taxa with specific immune-modulatory properties.

Differences in major bacterial populations in the intestines of mature broilers after feeding virginiamycin or bacitracin methylene disalicylate

The purpose of this study was to compare the effects of feeding virginiamycin or bacitracin methylene disalicylate (BMD), two in-feed antibiotics typically used by commercial poultry producers in the United States, on the chicken gastrointestinal microbiota. 454 pyrosequencing of the V6-V8 region of the 16S rRNA gene and quantitative PCR were employed to examine the bacterial microbiota and Clostridium perfringens, respectively, in the jejunum and caecum of market-age broiler chickens over four replicate grow-outs. Our results suggest that virginiamycin has a more pronounced impact on broiler gastrointestinal tract bacterial communities, relative to BMD, manifested primarily through significant enrichments in the genus Faecalibacterium in the caecum and a distinct population of Lactobacillus, OTU_02, in both the jejunum and caecum. No evidence for a difference among the diets in Cl.perfringens levels in the jejunum or caecum was observed. This work represents the highest resolution comparison to date of the jejunum and caecum microbiota in broilers fed either virginiamycin or BMD, and provides evidence for specific bacterial OTUs potentially involved in the health and performance benefits typically attributed to these in-feed antibiotics.