云南地区四种病原体在小兽类宿主中的感染特征分析-analysis of infection characteristics of four pathogens in small mammals in yunnan.docxVIP

5 5 mammalian reservoir hosts in Yunnan province Abstract Objectives The study aimed to investigate the infection status, the host constitute and genetic evolutionary characteristics of Bartonella, Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Spotted fever group rickettsiae in the small mammal hosts fromYunnan Province in order to provide the basis for the prevention and control of the four natural foci of disease pathogens. Methods Samples were selected from 32 sampling points in 12 counties of 7 cities (prefecture) in the western area of Yunnan Province with elevational ranges from 900 meters to 4300 meters at north latitude 20 °-30 °. Folder night method and trapping method were used to collect small mammal hosts.Studies were carried out to identify the type, gender, record latitude, altitude, vegetation type, habitat, body length and types of the rodents, and the spleens were collected . According to kit operating steps, total-DNA from spleen samples was extracted. Specific primers of Bartonella, Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Spotted fever group rickettsiae using to amplify the groEL gene , 5s-23s rRNA gene, 16s rDNA gene and 17-kDa gene were used in Nested-PCR. The PCR products were sequenced by sequencing company. All statistical analyses were performed with the SPSS 17.0 and SAS 9.1.3 statistical software package. χ2-test, Fishers exact test and Poisson regression models were PAGE PAGE 6 used.Phylogenetic trees were constructured based on the sequences of groEL gene , 5s-23s rRNA gene, 16s rDNA gene and 17-kDa gene, using the neighbor-joining method with Kimuras two-parameter distance method by MEGA 5.1. Bootstrap analysis was carried out with 1,000 resamplings. Results A total number of 2310 small mammal hosts were collected from Yunnan province during 2005-2006 and 2009-2011 with 11 types of Species included. Among all the samples, 412 samples were detected as positive for Bartonella with the p