Mucosa-associated microbiota that is intestinal plays a critical biological role due to its contiguity with the animal host.


Effect of High Grain Diet on Mucosa Microbiota and Colonic Mucosal injuries in sheep.


Background Information
Mucosa-associated microbiota that is intestinal plays a critical biological role due to its contiguity with the animal host. Scientific knowledge of its genetic composition is limited. The microbial communities are linked mainly with the duodenum, rumen, and colon of sheep (Collado and Sanzy 299). The gastrointestinal tracts of sheep can be investigated using cultural and hybridization techniques (Petri et al.). Significant levels of Lactobacillus will be identified in the duodenum and rumen to substantiate the presence of mucosa-associated microbiota. The agricultural modern ruminant productions manly feed the animals on high grain (HG) diets to increase milk output and register high efficiency (Wang et al. 1). Epithelium and microbiota of the gastrointestinal (GI) are the primary challenges posed by the feeding procedures. Increased microbial fermentation in the hindgut causes physical and immunological barriers. It is therefore essential to use advanced DNA and laboratory technologies to acquire more scientific knowledge concerning the genetic alignment of microbiota that is mucosa-related and how it relates to induced mucosal injuries.

Role of DNA in the study
The drawing out of mucosa’s DNA from the colon plays a critical role in gaining genetic insights about the biological compositions of all the bacteria involved in the fermentation process in the hindgut. The genetic data are also crucial in understanding the evolution of the microorganisms during different feeding and veterinary medication cultures. It is possible to know how starch presence in the colon plays a role in the fermentation and development of the bacteria. This paper aims to use Polymerase Chain Reaction (PCR) study the active modifications in microbiota in the colon of sheep by use of the genetic samples that are linked to the mucosa.
PCR Summary
The test-based investigation was permitted and executed in Nanjing Agricultural University with the help of The State Science and Technology Commission. It involved a total of six male sheep with a mean weight of 25.6 kg. They were isolated in four groups, and three of the classifications were supplied with feeds containing large amounts of grains for seven, fourteen, and twenty-eight days respectively. One group consumed pure hay diet during that period. Sections of digesta from the alimentary canal were taken to investigate the levels of acidity of fats. Microbiological classes were categorized with the use of mucosa samples from the colon. A microbial analysis, DNA extraction, and investigation of cytokines were aimed at finding out the effects of HG feeding on the animals.
The microbial DNA extraction of the mucosa was taken from the colon (Wang et al. 3). The retrieval was executed after craping the colonic mucosal samples with the help of sterile slides at a storage of -80 degrees Celsius. A QIA DNA animal waste kit was operated in administering the process that involved 0.26 grams of mucosa from the animals’ digestive system. Zirconium beads and Inhibit buffers were mixed with the mucosal sample and heated at 95 degrees Celsius to escalate the production of DNA. The DNA was stored at -80 degree Celsius for further examination. The percentage of starch from the digestive substance from the animal’s system were analyzed.
The PCR amplification utilized the V3 to V4 region of the extracted bacteria. 338F and 806R were the gene amplification the 16S rRNA. In the initial stage of the process, the extraction was denatured at a high temperature. Sequencing banks were constructed using the Amplicons. Linearization, isothermal amplification, and hybridization were then administered. The amplification was specific to the V3-V4 regions of the bacteria because correlation tests could still be carried out even after denaturing. A correlation examination was applied to investigate the relationship between mucosal-associated microbiota and colonic inflammatory cytokine.
The data results from the laboratory tests for the animal sample that were supplied with hay and HG for different weeks were taken and recorded (Wang et al. 4). VFA concentrations, starch contents, Lactate and Valerate, and pH concentrations were retrieved among other details as shown in the figure below.

The data was used to carry out a principal coordinate analysis that showed an existence of communities of bacteria in the mucosa extracted from the animals feeding on hay. It is clear from the statistical analysis that HG diet full of starch affects the abundance of colonic mucosa. The results also substantiated an unhealthy relationship between mRNA of the samples and the colonic Hydrogen potential. Feeding HG undoubtedly reduces the functionality of microbiome in the sheep’s digestive track. Different dynamic shifts according to the data analysis indicated the relationship between mucosal-associated microbiota and the consumption of HG diet by the sheep.
Throughout the process of adapting to the HG diets, the bacterial community linked to colonic mucosa shifted dynamically (Wang et al. 8). The data indicated that starch consumption from HGs raised the levels of some bacteria in the colon. The research also confirmed previous studies that reported a decisive role of gut bacteria in marinating the health of the host (Tajima et al. 273-284). A slough on the surface of the epithelium layer causes mucosal injuries to the animal. The severity of the quagmire is escalated by changes in fermentation processes in the colon and increase in the bacterial percentage. In conclusion, the starch that flows into the hindgut due to HG feeds alters microbial fermentation resulting in increased pH and higher levels of VFA in the digesta of the colon. The functions of the colonic mucosal bacteria are modified by increased colonic fermentation that is escalated by HG feeds. It is essential to use more extended periods for such research in the next steps of investigating mucosa-associated microbiota because adaptation to HG takes time.

Works Cited
Collado, MC and Y Sanzy. “Quantification of mucosa-adhered microbiota of lambs and calves by the use of culture methods and fluorescent in situ hybridization coupled with flow cytometry techniques.” Vetrinary Microiology (2006): 299-306. Document.
Petri, R, et al. “Characterization of the core rumen microbiome in cattle during transition from forage to concentrate as well as during and after an acidotic challenge.” PLOS ONE (2013). Document.
Tajima, K, et al. “Rumen bacterial community transition during adaptation to high-grain diet.” Anaerobe (273).
Yue Wang, Lei Xu, Junhua Liu, Weiyun Zhu and Shengyong Mao. “A High Grain Diet Dynamically Shifted the Composition of Mucosa-Associated Microbiota and Induced Mucosal Injuries in the Colon of Sheep.” Dynamics of Colonic Microbiome (2017): 1-13. Document.