Investigation And Analysis Of Erhlichia And Anaplasma Pathogen From Sheep And Wild Rodents In Parts Of Xinjiang

Anaplasmosis and ehrlichiosis are new zoonosis showed globally distributed,which pathogens belong to The Anaplasma Division,Rickettsia Head, and they were both Intracellular parasites spread by tick,rodents, ruminant and human can become the host.. Because their clinical symptoms were very similar to virus infection, there were often misdiagnosed and didn’t arouse enough attention. With the warming of the global climate, the ticks activated increasingly frequent which caused these diseases widely spread and increased incidence. In2008,《Human granulocytic anaplasmosis Control and Prevention Technical Guide (Trial)》 notice was promulgated by Ministry of Health of China, supervised subordinate departments for the prevention and control work. Xinjiang is located in the Asia-Europe continental central, Northwest China, borders with Kazakhstan, Russia and other countries. With diversified geographical environment and climate Xinjiang is an important pasture of our country. At present, there were few reports about Anaplasma infection in Xinjiang. Sheep are the main livestocks in Xinjiang region and mainly to grazing, the flow of the flock is bound to aggravated the spread of pathogens. The diversity of climatope in Xinjiang provided a natural habitat to the rodents which also aggravated the spread of pathogens, constituted a great threat to pastoral people’s public health and safety. So the research of rodents and sheep carried Anaplasma pathogens conditions and nosetiology is very necessary.Isolation of Anaplasma pathogen is a difficulty in world,although this pathogen was isolated successfully in some laboratory abroad, because isolation of pathogen was difficult,so there was little isolates of our country.To investigation the rodents of some districts in Xinjiang infected Erhlichia and Anaplasma,445rodents was captured in Bole Shihezi Urumqi and Yili, include356gerbils,58sewer rats and31field mice. Extract DNA from spleen of these rodents, and use nest PCR to amplify fragment of Erhlichia and Anaplasma16SrRNA gene, the positive PCR products was sequenced, the sequences was compared with the corresponding sequences deposited in GenBank. Among these samples, only gerbils found positive samples, these samples was identified as Erhlichia chaffeenisis and Anaplasma phagocytophilum after analysed to the sequence. Among the356gerbil samples,50showed Erhlichia positive, in which the rate is14.04%;17showed Anaplasma positive, in which the rate is4.94%,13showed both positive, in which the rate is3.79%. Results indicated that infection of Erhlichia and Anaplasma was exited in Xinjiang, most host was gerbil which had liven in desert and semi-desert. Some districts in Xinjiang are the origin place of these tow pathogens,and the two pathogens can exist in the same host.192sheep blood samples were collected from a sheep farm in Manasi and109sheep blood samples were collected from a shambles in Shihezi,.Total DNA was extracted from blood sanples which collectived from sheep and nest PCR targeted16SrRNA gene,The sequences of16SrRNA gene fragments from positive sample were compared with corresponding Anaplasma sequences deposited in GenBank.23anaplasma positive and77ehrlichia positive were found in192samples which collected from Manasi,the rate was11.97%and40.10%represently, after sequence analysed, these two fragments has only2basic different from rodent fragments which obtained in this study.37anaplasma positive was found in109sheep samples which collected from Shihezi by PCR used sheep anaplasma specific primers, the rate was34%, the similarities of the sequences amplified to the the corresponding sequences in GenBank were96%-99%.It is concluded that the infection of Anaplasma and exist in sheeps in Xinjiang province., and Further confirmed Xinjiang is the origin place of tow pathogens.Collected sheep blood which showed both Erhlichia and Anaplasma positive detected by PCR inoculate experiment mice which showed both Erhlichia and Anaplasma negative though intraperitoneal injection and HL60cells which growed best. Collected wild rodents’s spleen, suspended by PBS, inoculate suspension to experiment mice which showed both Erhlichia and Anaplasma negative though intraperitoneal injection and HL60cells which growed best. Collected positive sheep blood to make blood smear and observed under microscope. Suspected inclusions morula-shaped can be seen. Positive experiment mice which inoculated were exist detected by PCR. HL60cells which inoculated showed only Anaplasma positve detected by PCR.This is the first amplified Anaplasma and Ehrlichia gene fragments from gerbils in Xinjiang, and is the first time to investigated epidemiology of Anaplasma and Ehrlichia exist in in sheep in Shihezi and Manasi, Xinjiang province, this study attempt to isolated and cultured two pathogens, used positive sheep blood as specimen infected HL60cells, laid the foundation for further research.