Comparative Analysis Of Uterine Bacterial Compostion In Healthy And Endometritis Buffaloes

Endometritis is one of the most common reproductive disorders in buffalo and results from uterine infection with bacteria following parturition, causing high economic losses due to decline in reproduction ability, and prolonged intercalving intervals. In order to provide the foundation for Endometritis prevention and treatment, we used culture-independent method to describe and compare the uterine bacterial composition in healthy and endometritis postpartum buffaloes. The results of my research were as follows:1. There were several advantages in using uterine douche to sample uterine fluid, such as being simple to operate, simple device and high qualitity sample.2. The DNA yield by QIAamp DNA stool mini kit+Bead beating method was significantly higher than the two others. Besides, the integrality, purity and readingness for subsequent analyisis of DNA extracted by QIAamp DNA stool mini kit+Bead beating method were good.3. Two16S rDNA clone libraries were constructed. Sequences were assigned to5major groups (Fusobacteria, Bacteroidetes, proteobacteria, Firmicutes and Actinobacteria). Most of the clone sequences form the endometritic status libraries (38.89%) were affiliated with the phylum Fusobacteria and all of these sequences had high identity (≧99%) with Fusobacterium necrophorum subsp. Several clone sequences were affiliated with the phylum Bacteroidetes (46.67%), Sequences phylogenetically related to Bacteroides heparinolyticus(55.95%) and Porphyromonas levii(42.86%). Sequences related to Firmicutes(7.72%) and Proteobacteria(6.67%) were found in minor proportions, but bacterium in these two phylum are relatively diverse. Sequences affiliated with Actinobacteria(0.56%) were detected. In the clone libraries from the healthy status groups, the sequences were affiliated with Fusobacteria (26.56%) and all of these sequences had high identity (≧99%) with Fusobacterium necrophorum subsp. Additionally,32.81%of the sequences were affiliated with the phylum Firmicutes, with approximately61.9%of the sequences showing high similarity(≧97%) with Clostridium cadaveris. Further,35.94%of the sequences were affiliated with the phylum Bacteroidetes, with approximately56.52%of the sequences showing high similarity (99%) with Bacteroides heparinolyticus. In addition to these co-dominant phyla, sequences from Proteobacteria (4.69%) appear less frequently. The main pathogens of endometritis was Fusobacterium necrophorum subsp、Bacteroides heparinolyticus and Porphyromonas levii. Clostridium cadaveris Porphyromonas somerae and Bacteroides pyogenes may be good for uterine health.