Assessment of the qualitative variation in bacterial microflora among compartments of the intestinal tract of dogs by use of a molecular fingerprinting technique.
ABSTRACT To evaluate the qualitative variation in bacterial microflora among compartments of the intestinal tract of dogs by use of a molecular fingerprinting technique.
14 dogs (similarly housed and fed identical diets).
Samples of intestinal contents were collected from the duodenum, jejunum, ileum, colon, and rectum of each dog. Bacterial DNA was extracted from the samples, and the variable V6 to V8 region of 16S ribosomal DNA (gene coding for 16S ribosomal RNA) was amplified by use of universal bacterial primers; polymerase chain reaction amplicons were separated via denaturing gradient gel electrophoresis (DGGE). Similarity indices of DGGE banding patterns were used to assess variation in the bacterial microflora among different compartments of the intestine within and among dogs. Bacterial diversity was assessed by calculating the Simpson diversity index, the Shannon-Weaver diversity index, and evenness.
DGGE profiles indicated marked differences in bacterial composition of intestinal compartments among dogs (range of similarity, 25.6% to 36.6%) and considerable variation among compartments within individual dogs (range of similarity, 36.7% to 579%). Similarities between neighboring intestinal compartments were significantly greater than those between non-neighboring compartments. Diversity indices for the colon and rectum were significantly higher than those of the duodenum, jejunum, and ileum.
Results indicated that the different intestinal compartments of individual dogs appear to host different bacterial populations, and these compartmental populations vary among dogs. In dogs, fecal sample analysis may not yield accurate information regarding the composition of bacterial populations in compartments of the gastrointestinal tract.
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ABSTRACT: El tracto gastrointestinal (GI) de animales contiene diferentes tipos de microorganismos conocido como la microbiota GI. Por mucho tiempo, la microbiota GI ha generado interés porque los microorganismos GI están involucrados en múltiples procesos fisiológicos en el hospedero, así perpetuando salud o enfermedad. Estudios recientes han demostrado que la microbiota GI de gatos y perros es tan compleja como en humanos y otros animales, revelado con el uso de tecnologías de secuencia modernas y otras técnicas moleculares. La microbiota GI incluye miembros de todos los tres dominios principales de vida (Archaea, Bacterias y Eucariotas), pero las bacterias son el grupo de microorganismos más abundante y metabólicamente activo. El estómago de gatos y perros esta principalmente poblado de Helicobacter spp., el cual en perros puede representar tanto como el 98% de toda la microbiota bacteriana en el estómago. El intestino delgado contiene una microbiota más diversa, conteniendo representantes de al menos cinco diferentes filos bacterianos (principalmente Firmicutes y Bacteroidetes). El intestino grueso contiene el grupo de bacterias más abundante (~1011 células bacterianas por gramo de contenido intestinal), diverso (al menos diez diferentes filos han sido detectados) y metabólicamente relevante en el tracto GI. La mayoría de las bacterias en el intestino grueso son anaerobios estrictos, los cuales dependen de la fermentación de sustancias no digeridas para subsistir. Aunque estudios recientes han dilucidado las complejidades de la microbiota GI en gatos y perros, más investigación todavía es necesaria para encontrar maneras de manipular exitosamente los microorganismos GI para prevenir y/o tratar enfermedades GI.Archivos de Medicina Veterinaria 12/2012; 45(2):111-124. · 0.35 Impact Factor
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ABSTRACT: BACKGROUND: The composition of the microbiota of the equine intestinal tract is complex. Determining whether the microbial composition of fecal samples is representative of proximal compartments of the digestive tract could greatly simplify future studies. The objectives of this study were to compare the microbial populations of the duodenum, ileum, cecum, colon and rectum (feces) within and between healthy horses, and to determine whether rectal (fecal) samples are representative of proximal segments of the gastrointestinal tract. Intestinal samples were collected from ten euthanized horses. 16S rRNA gene PCR-based TRFLP was used to investigate microbiota richness in various segments of the gastrointestinal tract, and dice similarity indices were calculated to compare the samples. RESULTS: Within horses large variations of microbial populations along the gastrointestinal tract were seen. The microbiota in rectal samples was only partially representative of other intestinal compartments. The highest similarity was obtained when feces were compared to the cecum. Large compartmental variations were also seen when microbial populations were compared between six horses with similar dietary and housing management. CONCLUSION: Rectal samples were not entirely representative of intestinal compartments in the small or large intestine. This should be taken into account when designing studies using fecal sampling to assess other intestinal compartments. Similarity between horses with similar dietary and husbandry management was also limited, suggesting that parts of the intestinal microbiota were unique to each animal in this study.BMC Research Notes 03/2013; 6(1):91.
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ABSTRACT: Disturbance of the beneficial gut microbial community is a potential collateral effect of antibiotics, which have many uses in animal agriculture (disease treatment or prevention and feed efficiency improvement). Understanding antibiotic effects on bacterial communities at different intestinal locations is essential to realize the full benefits and consequences of in-feed antibiotics. In this study, we defined the lumenal and mucosal bacterial communities from the small intestine (ileum) and large intestine (cecum and colon) plus feces, and characterized the effects of in-feed antibiotics (chlortetracycline, sulfamethazine and penicillin (ASP250)) on these communities. 16S rRNA gene sequence and metagenomic analyses of bacterial membership and functions revealed dramatic differences between small and large intestinal locations, including enrichment of Firmicutes and phage-encoding genes in the ileum. The large intestinal microbiota encoded numerous genes to degrade plant cell wall components, and these genes were lacking in the ileum. The mucosa-associated ileal microbiota harbored greater bacterial diversity than the lumen but similar membership to the mucosa of the large intestine, suggesting that most gut microbes can associate with the mucosa and might serve as an inoculum for the lumen. The collateral effects on the microbiota of antibiotic-fed animals caused divergence from that of control animals, with notable changes being increases in Escherichia coli populations in the ileum, Lachnobacterium spp. in all gut locations, and resistance genes to antibiotics not administered. Characterizing the differential metabolic capacities and response to perturbation at distinct intestinal locations will inform strategies to improve gut health and food safety.The ISME Journal 02/2014; · 8.95 Impact Factor