Epidemiology Investigation Of Intestinal Parasites Of Sheep And Goat In Anhui Province And The Surrounding Areas

To determine the intestinal parasite infections of sheep and goat in Anhui province and surrounding regions,eight hundred and thirty-two sheep and seven hundred and eighty-one goat fresh fecal samples were collected from seven large-scale sheep farms and 10 large-scale goats farms in Anhui province and surrounding regions as Henan province,Jiangsu and Shandong province and examined the presence of oocysts or eggs using the centrifugation method,the Lugol'iodine-solution staining method,the Sheather's sugar flitation and modified acid-fast method.The results showed that the total infection rate were 47.60%and 62.87%in sheep and goat respectively.There were Eimeria spp.,Strongylus spp,Trichuris,Nematodirus,Moniezia,Giardia,Amoeba and B.honminis were found in sheep.Among them,the Coccidium and Amoeba were dominant in sheep with the infection rate 22.60%and 17.55%,respectively.The Eimeria spp.,Strongylus spp.,Trichuris,Nematodirus,Moniezia,Giardia,Amoeba,Paramphistomum,Trichuris and Cryptosporidium were found in goat.Among them,the Coccidium and Strongylus spp were dominant in goat with the infection rate 32.39%and 37.39%,respectively.Most of which were mixed infections and infection intensity was also larger.The most common infection in sheep the mixture of coccidia and Amoeba with the mixed infection rate 33.63%.The The mixed infection rate(91.67%)was highest in Tai'an,Shandong province.The most common infection in goat the mixture of coccidia and Strongylus spp with the mixed infection rate 51.72%.The The mixed infection rate(70.82%)was highest in Luoning,Henan province.The Coccidian and Strongylus spp were found with the largest infection intensity,with the OPG(EPG)reaching to 1020 and 3640,respectively.To determine the prevalence and molecular characterization of Cryptoridium isolates from sheep and goats in AnHui Anhui province and surrounding regions,DNA was extracted from the all fresh fecal samples from sheep and goat.all specimens DNA were detected using a nested-PCR targeting SSU rRNA gene of Cryptosporidium.On the basis of analysis of SSU rRNA,the gp60 gene from all the SSU rRNA-positive sample were amplificated and sequenced to indentify the gene subtype of Cryptosporidiu.The results showed the infection rate of Cryptoridium was 5.77%in sheep,and the infection rate in wujiang,Jiangsu province and Bengbu,Anhui province were 49.33%and 3.19%,respectively.There was no Cryptoridium infection in other sample regions.The infection rate of Cryptoridium was 8.71%in goat,and the infection rate was 46.00%,10.09%,and 7.64%in Maanshan,Anqing,and Fuyang,Anhui paovince.There was no Cryptoridium infectionin goat in other sample regions.The infection rate of Cryptoridium was 8.71%in goat,and two Cryptosporidium species including C.xiaoi(27)and C.ubiquitum(21)were identified in sheep,and C.parvum(68)was only identified in goat.The gp60 gene was successfully sequenced in C.ubiquitum(19)and C.parvum(54)isolate,with XIIa for C.ubiquitum and IIdA19G1 for C.parvum.To determine the prevalence and genotype characterization of E.bieneusi isolates from sheep and goats in Anhui province and surrounding regions,All specimens DNA from sheep and goats were detected using a nested-PCR targeting ITS gene of E.bieneusi,and were genotyped by bioinformatics methods.The results showed the infection rate of E.bieneusi was 3.37%(28/832)in sheep,and E.bieneusi was positive in Bengbu(2.0%),Maanshan(4.0%),Wujiang(21.33%)and Taian(1.67%).The infection rate of E.bieneusi was 4.09%in goat,and E.bieneusi was positive in Chuzhou(19.09%),Bengbu(11.11%),and Xuzhou(9.86%).DNA sequence analysis of E.bieneusi-positive specimens showed bioinformatics analysis showed genotype COS1,OEB1,BEB6,AHS1,AHS2 and SZS1 were found in sheep,genotype CHG1,CHG3,AHG2 and BEB6 were found in goat.Among them,AHG1,AHS1,AHS2 and SZS1 were new genotype in this discovery.All genotypes obtained in this study belong to Group 2.To determine the prevalence and molecular characterization of G.lamblia isolates from sheep and goats in Anhui province and surrounding regions,all specimens DNA from sheep and goats were firstly detected using a nested-PCR targeting TPI gene of G.lamblia,then the TPI-positive samples were analyzed by a hemi-nested-PCR targeting the GDH gene of G.Lamblia.The TPI and GDH PCR products were sequenced and analyzed using bioinformaticts methods to identify the G.lamblia assemblages.The results showed the positive rates of G.lamblia infection in the 832 sheep fecal samples and 781 goat fecal samples were 0.36%(3/832)and 0.0%(0/781),respectively.The two GDH genes,identified as assemblages E,were successfully amplicated and sequenced in three TPI-positive isolate.The TPI gene sequences were identified as assemblages E1(n = 2)and E6(n ?1).To determine the prevalence and molecular characterization of B.hominis isolates from sheep and goats in Anhui province and surrounding regions,all specimens DNA from sheep and goats were detected by PCR based on the "barcode region" of the small subunit ribosomal RNA(SSU rRNA)gene of B.hominis.The sequenced obtained were submitted to B.hominis 18s database to determine its genotype and 18s allele.The results showed the positive rates of B.hominis infection in the 832 sheep fecal samples were 6.00%(50/832).Among then,the positive rates of B.hominis infection in the Wujiang,Jiangsu province,Taian,Shandong province and Bengbu,Anhui province were 24.00%(18/75),16.67%(10/60)and 6.38(22/345),respectively.Other regions was not found infection.The positive rates of B.hominis infection in the 781 goat fecal samples were 0.26%(2/781),and only found B.hominis-positive sample in Maanshan,Anhui province with 2.00(2/100).Sequence analysis showed that the sheep carried ST4(2),ST5(8),ST10(25),ST14(10),new 1(3),new 2(1)and new 3(1),and the goat carried ST1(2).The 18s allele analysis showed that the sheep genotype ST4,ST14,ST 10 and goat genotype ST1 only posses one allele as allele 2,allele 42,allele 19 and allele 43,respectively.The sheep genotype ST5 have two alleles as allele115 and allele119.To determine the prevalence and molecular characterization of P.hominis isolates from sheep and goats in Anhui province and surrounding regions,all specimens DNA from sheep and goats were detected and genotyped by a nest-PCR technique targeting ITS gene of P.hominis.The results showed that P.hominis was negative in sheep.The positive rates of P.hominis infection in goat was 0.26%(2/781),and two P.hominis-positive samples were located in Luoyang,Henan province(1.83%).Sequence analysis of ITS gene showed that positive isolate belong to genetype CC1.