Preliminary Study On The Function Of Carrying And Spreading Microorganisms By Wild Birds

Objective: In order to explore the function of carrying and spreading microorganisms through the body surface of wild birds,this study combines high-throughput technology with pure culture methods to detect microbial groups from the surface of wild birds,comparing and analyzing the differences of the carrying model on microorganisms among different ecological wild birds.The object of this study is to understand the spatial distribution pattern of microbial diversity from the perspective of carrying and spreading microorganisms by wild birds.Method: The wild birds were captured by the light trap and fog net methods,and 391 samples were collected aseptically.Among them,12 samples of 5 species of wild birds' claws were used for high-throughput sequencing to detect microorganisms;379 samples of 35 species of wild bird faeces and cotton swabs from beak,claw,wings,tails samples were used for pure culture to detection of Nematode-Trapping Fungi(NTF).High-throughput sequencing technology based on Amplicon sequence variants(ASV)and the Illumina Nova Seq platform was used to amplify the variable regions V3 and V4 of 16 S r RNA of bacteria,and the variable regions ITS1-1F of ITS of fungi.The monosporium separation method was used to isolate and culture NTF from the sample,and species were identified by combining morphology and molecular technology.Qiime2 software was used to carry out quality control,pruning,noise removal and splicing on high-throughput sequencing data,and Excel software was used to analyze pure culture results.Results:(1)The high-throughput sequencing of the microorganisms carried by the wild bird claws did not detected fungi but only detected bacteria.The bacteria were annotated to 27 phylum,62 classes,105 orders,163 families,255 genus and 133 species.The top five detected phylum were as follows Firmicutes,Proteobacteria,Bacteroidetes,Actinobacteria,Fusobacteria,and the content of that detected accounts for 96.68%-97.60%.The top ten detected genus were Campylobacter,Ralstonia,Streptococcus,Actinomyces,Prevotella,Rothia,Veillonella,Ruminococcus,Prevotella and Unspecified,and the content of that detected accounts for 35.43%-53.29%.The bacterial community composition and diversity on the claws of different wild birds are obviously different.Alpha diversity and Beta diversity analysis were carried out on bacterial communities of different bird claw samples,and the results show that the diversity and richness of bacterial community on water birds are the lowest,and the similarity of bacterial community among forest birds is higher than that between forest birds and water birds.In addition,at the level of genus and species,the total proportion of pathogenic bacteria carried by wild birds is quite high,count for 81.25% and75.00% respectively;and the composition of pathogenic bacteria carried by forest birds and water birds are different,with that the pathogenic groups carried by forest birds are richer than water birds.(2)A total of 9 NTF strains were isolated based on pure cultured samples,the detected parts all were claw and the detection rate was 8.7%,including Arthrobotrys oligospora,Arthrobotrys conoides,Arthrobotrys thaumasia and Arthrobotrys multiformis.From the prospective of ecological group,the NTF detection rates of forest birds and water birds respective were 6.9% and 10.9%,the NTF detection rate of water birds were much higher.From the residence status of birds,the NTF detection rates of migrating process birds,resident birds and arrive destination birds respective were 3.6%,12.5% and 15.0%.The detection rate of NTF on resident birds and arrive destination birds is much higher than that of migrating process birds,and the detection rate of NFT on resident birds and arrive destination birds are not much different.Conclusion:(1)The body surface of wild birds can carry dominant bacterial groups and widely distributed NTF groups,which can be used to explain the formation of widely distributed NTF spatial distribution pattern;(2)The ecological habits of wild birds affect the diversity of microbial species carried by them: the diversity of pathogenic bacteria carried by forest birds is higher than that of waterbirds.The NTF detection rate carried by water birds is higher than that carried by forest birds.The detection rates of NTF resident birds and arrived destination birds are much higher than that of migrating process birds;(3)Pathogenic bacteria carried by forest birds have not been considered seriously,and the monitoring of surface microorganisms of forest birds needs to be included in the daily monitoring scope of Epidemiology.