Diversity Of Symbiotic Actinobacteria From Healthy Ruminant Faeces

In order to improve the efficiency of the screening of natural products,international scientists use functional genome research outcomes to improve or designnew screening system, selective isolation of actinomycetes from natural ecologicalsystem with new technology is also launched, including isolation of rareactinomycetes in particular attention. There aren’t much study on Animal symbioticActinomyces at present, while using traditional and molecular ecology technique tostudy diversity and function of them, we can not only find new Actinomyces speciesproviding resources for the screening of natural products, but also get scientificunderstanding of antibiotic effects on animal growth mechanism which lay solidfoundation for the development of new probiotics products.In the study, we collected the feces samples of cow and sheep from Henanprovince.The actinomycetes communities were investigated by culture and cultureindependent molecular approaches．Results from our studies were summarized asfollows:1. The culture dependence methods were used to isolate actinomycetes from thesesamples. The phylogenetic analysis indicated that the sequences belonged toStreptomyces、Micromonospora、Amycolatopsis and Cellulosimicrobium.2. Two16S rDNA gene libraries were constructed and analyzed by amplified ribosomal DNA restriction analysis technique(ARDRA)．The clones of dominantARDRA patterns were selected to sequence．The phylogenetic analysis indicated thatthe sequences from dorninant ARDRA patterns of cow mostly belonged toMycobacterium、 Rhodoglobus and Nocardioides, while the sequences fromdorninant ARDRA patterns of sheep mostly belonged to Aeromicrobium、Streptomyces and Nocardioides.3. All isolated strains were tested for antimicrobial activity using agar diffusionmethod.The result showed that there are39antimicrobial actinobacteria strains of79actinobacteria from cattle faeces sample,11showed activity against Escherichiacoli,which was11.3%of the total isolates.5showed activity against Bacillus subtilis,which was5.16%of the total isolates.6showed activity against Candida Albicans,which was6.19%of the total isolates.6showed activity against Rhodotorula, whichwas6.19%of the total isolates.7showed activity against Saccharomyces cerevisiae,which was7.22%of the total isolates.However, only4strains showed activity againstMycobacterium,which was4.12%of the total isolates.While65antimicrobial actinobacteria strains of139actinobacteria from sheepfaeces sample.26showed activity against Escherichia coli,which was18.7%of thetotal isolates.5showed activity against Bacillus subtilis, which was3.6%of the totalisolates.6showed activity against Candida Albicans, which was4.32%of the totalisolates.10showed activity against Rhodotorula, which was7.19%of the totalisolates.13showed activity against Saccharomyces cerevisiae, which was9.35%ofthe total isolates.However, only5strains showed activity againstMycobacterium,which was3.6%of the total isolates.Accrding to the diameter of antibacterial circle, bacteriostatic ability of culturedependent actinomycetes from sheep faeces sample is stronger against Escherichiacoli、Candida Albicans、Saccharomyces cerevisiae、Mycobacterium, Bacteriostaticability of culture dependent actinomycetes from cattle faeces sample is strongeragainst Bacillus subtilis, while Bacteriostatic ability of culture dependentactinomycetes from both samples is almost the same against Rhodotorula.4. We selected actinobacteria S161, S154, C244and C272which have strong antibacterial ability for metabolomics studies, preliminary metabonomics study resultscan be divided into two types, one is C272strain metabolic pattern whose majormetabolic molecular weight is between700and800; the other is C244strainmetabolic pattern whose major metabolic molecular weight is1157. The conclusion isthat the metabolic fractions of nonvolatile compound components are not rich, buthigh molecular weight is their common characteristics.