| Anaplasmosis is a kind of tick-borne disease,caused by Anaplasma,which parasitizes in host animals’ blood cells.It is widely distributed all over the world,and can infect many kinds of animals.For example,wild mammals,birds,ruminants,rodents,reptiles,domestic animals and human can be infected by Anaplasma phagocytophilum,a kind of pathogeny threatens the development of breeding industry and human health,so it is a kind of zoonosis with great significance of human public health security.However,now adays,the studies of Anaplasma’s hostmostly focus on rodents and some of ruminants.Therefore,the infection of different animals in some areas of China was researched in this study and can provide some scientific basics to the prevention and control of this disease.1.To know the epidemic situation of Anaplasma in blood among different hosts in some areas of China,and provide some scientific basics of the determination of hosts in these areas,285 samples from goat and sheep blood in Henan province(Yiyang and Xinan countries in Luoyang,Ruzhou,Changge and Hebi),Guizhou province(Maiping,longli and Pan countries),Yunnan province(Longling country in Baoshan),240 samples from dog blood in Zhengzhou,Yiyang and Xinan countries in Henan province,and Maiping country in Guizhou province,44 samples from cattle blood in Hebi,Henan province,and 103 samples from wild bird blood and 16 samples from wild mammals in Zhengzhou Zoo were tested to detect three kinds of Anaplasmas by nest-PCR from March 2014 to January 2016 in this study.The result is that three kinds of Anaplasmas were detected from blood of all these animals,and the prevalence differd in different hosts.The prevalence differd in different areas,prevalence of Anaplasmas form Henan province was 77.32% and 91.67% in Guizhou and Yunnan province respectively,which was higher than the total rate in Henan,speculated relating to the different environments.The prevalence was different in various hosts,goat and sheep was 54.74%(156/285)infected,in which A.phagocytophilum contained 23.86%(68/285),A.bovis contained 43.86%(125/285),and A.ovis contained 23.86%(68/285).The infection of three kinds of Anaplasmas was found firstly in dogs,the prevalencein dogs of A.phagocytophilum,A.bovis and A.ovis were 0.42%(1/240),2.08%(5/240)and 5%(12/240)respectively.All kinds of Anaplasmas were firstly detected in wild animals including giraffe,red deer and fox.The prevalence of sheep and goat was higher than other hosts,which may relate to the host specificity of different variants.The prevalence of Anaplasmas in goat was 76.19%(122/147),which was far above sheep’s 24.64%(34/138).The prevalence in grazing sheep and goat was 84.15%(138/164),which was significantly higher than the hand-fed ones’ 14.88%(18/121).The prevalencein dogs raised in sheep farms was higher than pet dogs raised in families.Mixed infection was universal,and the rate was 30.18%(86/285).2.In order to understand the differences between three kinds of Anaplasmas,samples were amplified based on the 16 S r RNA of A.phagocytophilum and A.bovis,the MSP4 gene of A.ovis.103 Anaplasmas positive samples were selected randomly from different areas and animals to be sequenced.Upstream and downstream sequences were aligned and spliced by Clustal X 2.11 and Vector NTI Advance 11.5.1.Then the homology between our sequences and reference sequence was analyzed by Meg Align of DNAStar 8.0.Phylogenetic of three kinds of Anaplasmas was analyzed by Mega6.06,and Maximum Likelihood method was used to analysis nucleotide sequence and to drawa phylogenetic tree.In this study the 16 S r RNA of A.phagocytophilum was detected from different animals’ blood,4 kinds of genotypes(A,B,C,D)were found from three kinds of animals(sheep,dog,and giraffe),in which 3 new genotypes hadn’t been reported yet.The genotype D differed much with other sequences,and the homology between them was 98.1%-98.2%.Genotype D had only 11 basesdifferent with the reference sequence KM186950,which is derived from sheep.New genotypes were detected from both dog blood and sheep blood,but there was only one base different with reference sequence,the homology was 99.8%.Whether AP isolate derived from sheep can affect dogs should be researched later.A.bovis16 S r RNA was found in different kinds of animals for the first time,and 5 kinds of genotypes were found in this study,genotype A was identical to the reference sequence KP314251,others(B,C,D,E)were newly found.Genotype E was firstly reported,the homology between it and other genotypes was the lowest,and 11 bases are different between genotype E and reference sequence.2 kinds of genotypes(A,B)were detected from sheep,and 3 were found in dogs,10 different kinds of A.ovis MSP4 were found in all hosts,in which only genotype A was identical to the reference sequence KJ782397,others all had some difference with the reported sequences,but with high homology.Genotype B,D,E,F,G,H of A.ovis MSP4 was firstly found in dogs,in which genotype B and G had the homology of 99.8% with FJ460443,which is derived from human,indicated that A.ovis MSP4 variant from dogs have the possibility to infect human.Genotype A,B,C,E were detected in sheep blood and genotype I and J were firstly found in foxes and red deer raised in zoo.This study fully indicated that the sequence of three kinds of Anaplasmas had genetic diversity.In this study,4 different genotypes of 16 S r RNA in A.phagocytophilum,5 different genotypes of 16 S r RNA in A.bovis and 10 different genotypes of MSP4 in A.ovis were found,and most of them hadn’t been reported before.In conclusion,this study provides some scientific basics and has some significance to the study related to zoonosis and the promotion of breeding industry. |
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