Molecular Biological Identification And Detection Of Partial Pathogens Of Melophagus Ovinus

Melophagus ovinus is a kind of ectoparasite with sheep as the main parasitic host,it can cause host anemia,loss of weight and wool and sheep skin quality and yield when it's parasitic on sheep,and cause direct economic loss.Moreover,M.ovinus can also spread or store many kinds of pathogens,such as Bartonella,Anaplasma ovis and Rickettsia,which pose a threat to animal husbandry and human health and cause indirect economic losses.M.ovinus were collected from Kuqa in Aksu,Aketao in Kizilsu Kirghiz Autonomous Prefecture,Qinghe in Aletai and Wuwei in Gansu Province.88(13 pupaes),5,5 and 10 add up to 108 samples with complete morphology were selected from four locations,through the observation of morphological characteristics,it is preliminarily identified as M.ovinus.Then the 12 S rDNA?16S rDNA and 18 S rDNA genes were amplified.And the polymerase chain reaction(PCR)was used to amplify the genes of Bartonella gltA?Arsenophnus 16 S rDNA?Anaplasma spp.msp4?Anaplasma ovis GroEL?Rickettsia ompA? Rickettsia 17 KDa and et al,and sequencing of positive products.On-line Blast analysis of all the sequences obtained from the experiment,the corresponding gene sequences were downloaded from the GenBank database.The homology and phylogenetic analysis were carried out by MEGA5.0 software to determine the kinds of these pathogens.The results show that,the similarity between the 12 S rDNA sequence of the M.ovinus and the 12 S rDNA sequence of the M.ovinus uploaded in Yunnan in 2016 is 100%;The similarity of 16 S rDNA sequence of M.ovinus with Denmark 2007,Czech Republic 2017 and Xinjiang 2017 upload 16 S rDNA sequence is 99%;The similarity between the 18 S rDNA sequence of the M.ovinus and the four sequences uploaded in Xinjiang in 2016 is 99%;The results of pathogens detection are as follows: The positive rate of Bartonella gltA gene was 95%(103/108),the homology with the known Bartonella melophagus gltA sequences ranged from 99.3% to 100%;The positive rate of Arsenophnus 16 S rDNA gene was 6.4%(5/78,all of the 5 DNA samples were derived from the pupae of M.ovinu),the homology with Arsenophnus 16 S rDNA detected in M.ovinu was 100%;The positive rate of Anaplasma spp.msp4 gene was 39%(42/108,among them,5 DNA samples were derived from the pupae of M.ovinu),the homology with the known A.ovis msp4 sequences ranged from 99.3% to 99.8%;The positive rate of A.ovis GroEL gene in was50%(39/78),the homology with the known A.ovis GroEL sequences ranged from 99.1% to 100%;The positive rate of Rickettsia ompA and 17 KDa genes were 9%(10/108),the homology with the known Rickettsia raoultii sequences were 99.4% ~ 100% and 99.7% ~ 100%,respectively;The positive rate of Wolbachia 16 S rDNA gene was 94%(102/108),the homology was 99.6% ~ 100% between the amplified products and those detected in many hosts,and further analysis show that it is Wolbachia supergroup F.Kuqa in Aksu,Aketao in Kizilsu Kirghiz Autonomous Prefecture,Qinghe in Aletai and Wuwei in Gansu Province were present M.ovinus,which mainly parasitic to sheep.It carries a variety of pathogenic microorganisms,suggested that animal husbandry should prevent M.ovinus and its possible harm to reduce the direct or indirect economic losses.