Isolation methods and diversity of culturable fecal actinobacteria associated with Panthera tigris tigris in Yunnan Safari Park

Abstract:

Abstract:［Objective］We studied the isolation methods and diversity of culturable fecal actinobacteria associated with Panthera tigris tigris by using culture-dependent approaches． ［Methods］Fresh fecal samples of healthy Panthera tigris tigris were collected from Yunnan Safari Park． Pretreatment of the samples，isolation media and inhibitors were tested for actinobacteria isolation． 16S rRNA genes of actinobacteria were sequenced and subjected to phylogenetic analysis．［Results］The abundance of culturable actinobacteria was 1.10×108 cfu/g colony forming units (CFU) per gram of feces (wet weight)．We obtained 110 purified cultural actinobacterium strains．The analysis based on 16S rRNA gene sequences showed that these strains were distributed in 10 different families and 12 genera of actinobacteria at least，and most of them were non-filamentous，such as Arthrobacter，Dietzia，Kocuria，Corynebacterium and Microbacterium．Streptomyces was the mainly classical filamentous actinobacteria，and up to 64% of total．［Conclusion］ Drying and heating up the fecal samples can greatly increase the rate of the actinobacteria．Many kinds of inhibitors and chemical defined media are suitable for isolation of fecal actinobacteria． The culturable actinobacteria are abundant in Panthera tigris tigris feces.Our study found an effective method to isolate animals' fecal actinobacteria and it's useful for studying and exploiting animals' fecal actinobacteria．