| The diversity of Giant Panda intestinal microflora is closely related to its health and it is unclear whether captive breeding influences gut flora composition.Captive breeding at the Chengdu Research Base of Giant Panda Breeding and the China Giant Panda Protection Research Centre produces the highest numbers of pandas in the world,but their panda management programmes are not standardised.We compared the intestinal microflora of pandas from the two research base to evaluate the effects of age,kinship and feeding management microbial community structre and diversity by using 16 S r RNA technology.We choose the ideal sample for the metagenomic analysis and analyzed the intestinal flora characteristics in four age groups of captive giant panda,to evaluate the effects of age and kinship.The main results were as following:1.Characteristics of captive-bred giant pandas intestinal flora vary among age groups,with familial relatedness and between different baseCharacteristics of captive giant panda intestinal flora between two breeding centres shown that giant panda’s intestinal flora is mainly composed of common bacteria,cellulose degrading bacteria,pathogens and beneficial bacteria,but the composition of captive panda’s intestinal flora between two bases is different.The abundance of cellulose degrading bacteria and beneficial bacteria were higher in pandas at Chengdu base,whereas in pandas at Wolong base,abundance of pathogens was higher.Relative abundance of microflora varied with age of pandas at both bases,we found that species clustering was higher at Chengdu base,reflecting the greater familial relatedness of the captive-bred pandas.We concluded that captive-bred giant pandas at Chengdu base may have a low probability of bacterial intestinal disease and a better likelihood of survival success on release to the wild,but the close familial relationships in the breeding programme should be avoided to improve genetic diversity.The result of sample detection in Chengdu base is ideal,which conforms to the requirements of the follow-up experiment.2.Captive giant panda intestinal microflora varies with age and kinshipUsing metagenomic analyses of fresh feces from infant,juvenile,adult and mature adult pandas,we found that the relative abundances of pathogenic(Streptococcus gallolyticus)and cellulose degrading bacteria(Clostridium sp.Marseille-P2414)increased with age,whereas those of probiotics(Lactobacillus murinus,L.johnsonii,Pediococcus acidilactici,and Bifidobacterium breve),carbohydrate and amino acid metabolism genes decreased.Cellulose degrading bacteria species varied among the age groups,and the relative abundance of immune function genes was lowest in juvenile pandas.We found that sibling and parent-child relationships affected microflora and gene clustering.We concluded that the composition of intestinal microflora and functional genes were distinct and different for each age group and causal factors may include diet;commonly occurring intestinal bacteria produce cellulose degradation enzymes when stimulated by diet,and Enterobacteriaceae play a regulatory role in the degradation of fiber;the relative abundances of carbohydrate and amino acid genes decreased with age,suggesting that the digestibility of bamboo by giant pandas decreases with age;relative abundance of immune function genes were lowest in juvenile pandas and could possibly increase the susceptibility of the age group to disease;and,the sibling and parent-child relationships influence the diversity of intestinal microflora in captive pandas.We recommend the introduction of genetic diversity into giant panda breeding programs.3.Identification and drug resistance detection of dominant bacteria in the feces of captive pandasUsing the traditional detection method to identify and detect the dominant bacteria of giant panda feces from two bases,58 strains of Shigella were isolated from from Chengdu samples,and 28 strains from Wolong samples.The results showed that Shigella was the dominant strain in the captive panda intestinal flora and these strains which isolated from two bases have high resistance to Augmentin and Cefazolin but were very sensitive to Amikacin.We concluded that the isolation rate of Shigella from two bases was different,the isolation rate of Chengdu base was higher than that of Wolong base,which accords with the abundance difference of captive panda intestinal flora in two bases;the drug resistance of the giant pandas in Wolong base is serious;the drug resistance is closely related to the drug introduction,and the sensitive drugs should be replaced regularly. |
