Analysis Of Microbial Diversity Of Intestinal Wild Animals In Nanchang District

In recent years,with the increasing range of human activities and the excessive deforestation and illegal hunting of wild animals,the habitat of wild animals has been destroyed and the number of many wild animals has drastically decreased.It is urgent to protect the survival and reproduction of wild animals and improve the living environment of wild animals.Therefore,more and more researchers have begun to pay attention to and research wild animals.However,there are relatively few studies on the intestinal microflora of wild animals.Most studies still use traditional culture methods,and the results are not comprehensive and accurate.High-throughput sequencing tecHNology has been widely used for the study of human and animal gut microbes due to its economy,accuracy,high sensitivity,and high throughput.This tecHNology can more thoroughly and accurately demonstrate the overall environment of the gut microbes.The extraction of high-quality total DNA from gut microbiota is the prerequisite for using high-throughput sequencing tecHNology to study intestinal microbes.The quality of total DNA extracted will affect the accuracy and veracity of follow-up experimental results,but there are huge differences in the quality of the total microbial DNA extracted by different methods.Therefore,it is particularly important to find a suitable method for extracting the total DNA of the gut microbiota.The specific content of this study are as follows:1.Comparative study on DNA extraction methods of feces from wild animalsIn this study,six extraction methods were combined with two extraction strategies,fecal total DNA of herbivorous and carnivorous wild animals was extracted at the same time.The results showed that method 1 directly extracted fecal microbial total DNA from carnivorous wild animals and method 3 plus pre-extracted herbivorous wild animal feces had higher total microbial DNA quality and could better reflect fecal microbial diversity.2.Analysis of intestinal microbial diversity of wild animalsA total of 20 fresh excrement samples from 10 wildlife were collected from the Nanchang Zoo.The samples were divided into herbivorous,meat,and omnivorous groups according to the wildlife's feeding habits.The herbivorous group was divided into rumination and monogastric herbivory groups according to the number of stomachs.A total of 1379320 sequences,with a total of 2,529 OTUs were obtained by using high-throughput sequencing tecHNology to sequence all the samples.The average number of OTU per sample in the meat,omnivorous,and herbivory groups respectively were 109.5±7.5,548.2±18.8,and 918.9±85.7;while average number of OTU in the ruminant group and the monogastric herbivore group respectively was 701±160.8 and 1064.2±33.7.Alpha diversity analysis results showed that this sequencing data better reflects the situation of the sample microorganisms,and can be used for the next analysis.Beta diversity analysis showed that the herbivore animal had the highest microbial diversity in the feces,followed by the omnivorous group,and the lowest in the meat group.In the herbivores,the microbial diversity in feces of monogastric herbivores and ruminants was somewhat different,and the fecal microbial diversity in the ruminant group was higher than the monogastric herbivore group.Statistical analysis was performed on the structure of the feces samples in each group of animals,the results showed that the predominant bacteria in the meat group were Firmicutes,Fusobacteria,Bacteroidetes and Actinobacteria;the predominant bacteria in the herbivorous group were Firmicutes,Bacteroidetes and Proteobacteria;the predominant bacteria in omnivorous group were Firmicutes,Bacteroidetes and Spirochaete.The predominant bacteria in the ruminant were Firmicutes and Bacteroidetes;the predominant phyla of the single stomach herbivores were Firmicutes,Bacteroidetes,Proteobacteria.The samples of each group were analyzed by difference analysis.,the results shown that the abundance of Bacteroidetes,Spirochaet and Verrucomicrobia in the herbivore faeces were higher than those in the carnivorous animals,while the abundance of Actinobacillus and Fusobacterian in the carnivorous faeces were higher than those in the herbivores animals;bacteriums in the phylum level were no significant difference from the faeces of the herbivore and omnivorous;Fusobacteria in the feces of the carnivore was higher than this in the faeces of omnivorous,while Spirochaete in the feces of omnivorous was higher than this in the faeces of carnivore;the Firmicutes and Tenericutes in the fecals of the ruminant were higher than those in the monogastric herbivore.The differences in metabolic function of each group of samples were analyzed.In this research,high-throughput sequencing tecHNology was used to study the microbial diversity of the intestine,and the community structure of the intestinal microflora of wildlife was roundly demonstrated.This provides a basic theoretical basis for further investigation of the mechanism of dietary and digestive physiology affecting intestinal microbial diversity and lays a scientific basis for better protection of wild wildlife.