Establishment and evaluation of multiplex PCR for detection of main pathogenic bacteria of endometritis in Tibetan sheep / 生物工程学报

A multiplex PCR method was developed to detect the main pathogens of Qinghai Tibetan sheep endometritis. First, the genomes of five standard bacterial strains were extracted and specific primers were selected; the multiplex PCR method was established by using the genome of the standard strain as a template. The samples were collected by sterile cotton swab from Tibetan sheep uterus, and then placed in LB medium and numbered. After 48 h, the genomes of cultured bacteria were extracted and detected by single PCR method, then the positive samples were recorded. The positive samples detected by single PCR were selected for multiplex PCR detection and recorded again. The coincidence rate between these two methods was calculated to measure the accuracy of multiplex PCR. In order to identify the species of the pathogen, 30 positive samples verified by single and multiplex PCR were randomly selected for bacterial isolation and identification. In the 600 samples, the infected ratio of Streptococcus agalactiae (GBS) was 47.33%, Escherichia coli 34.83%, Staphylococcus aureus 6.5%, Salmonella and Trueperella pyogenes were negatively detected. Among the positive samples detected by multiplex PCR, the positive ratio of GBS was 45.50%, E. coli 33.50%, S. aureus 6.5%. Comparison of two detection results, Multiplex PCR detection coincidence rate is more than 95%. The isolated pathogens were identified as E. coli, GBS and S. aureus, which was consistent with the results of two methods. The multiplex PCR method was successfully established and the main pathogens of endometritis in Qinghai Tibetan sheep were GBS, E. coli and S. aureus.

OBSERVATION OF ADULT YAK PINEAL GLAND BY LIGHT AND ELECTRON MICROSCOPY / 解剖学报

Objective To study the characteristic of histological structure of the pineal gland of the adult yak. Methods HE and Achucarro-Hortega staining,light microscopy and transmission electron microscopy were used. Results The pineal gland was composed of pinealocytes,occasional glial cells,capillaries and neural elements.The pinealocyte appppered clear electronic density and its cytoplast included numerous mitochondria,rough and smooth endoplasmic reticulum,microtubules,microfilament and ribosome.Golgi complex was rare.The typical heterogenous organelle of pinealocyte of yak was spherical synaptic ribbon,which located near the plasma membrane.The glial cells contained abundant of mitochondria,and their processes which appeared bulbous invested the pineal periphery and incompletely separated the pinealocytes.Synapse and junctional complex existed between the pinealocytes and glial cells.Capillaries in pineal gland of the yak were belong to continuous capillaries,and pigment cells were observed around the vascellum in the gastr-distally of the pineal gland. Conclusion Neuroepithelial and epithelioglandular cell conjuction can be observed in the pineal gland of adult yak.Capillaries are belong to continuous capillaries.There are aboundant of organelle except Golgi complex in the pinealocyte.