| Amebiasis,Giardiasis and microsporidiosis are important zoonotic protozoan disease,and their organisms can infect human and many animals.Amebiasis,caused by Entamoeba histolytica,can induce amebic dysentery and liver abscesses.Approximately more than 10%of the population have been reported to be infected with this disease from various countries,resulting in 100000 deaths each year.Now,in developing countries,it is still one of the important reasons causing infection and death.Giardiasis is caused by Giardia spp,of which Giardia.duodenalis can infect human,many farm animal,companion,aquatic animal and wild animal,mainly leading to diarrhea,abdominal carmps,bloating,weitht loss,malabsorption,and even death.Enterocytozoon bieneusi can induce self-limited diarrhea of immunocompetent persons and many animals,and even death-leading diarrhea for AIDS patients.Many zoonotic E.bieneusi genotypes haved been found in wastewater,except in human and animals.The potential possibility exsits in transmission between animals and human,because the intimate contact between human and farm animal,companion animal and wild animal,thus learning the epidemiology of these three organisms has important public health significance.1.The epidemiology of Entamoeba spp.from nonhuman primatesNonhuman primate is the main experimental animal in medicine research field,in order to study the infection,geographic distribution and zoonotic risk of Entamoeba in nonhuman primate from different district and with different living style,2688 nonhuman primates' fecal samples were collected from monkey farm,zoo,park and Natural Scenic Area(free living),in some part of Sichuan,Henan,Yunnan,Guangdong and Guangxi.1059 were positive for Entamoeba by microscopic examination,and the infection rate was 39.40%(1059/2688).The Entamoeba infection exsited in all the districts,of which the infection rate of Guangdong was highest,53.03%,and infection rate of Sichuan is lowest,25.38%.According to the result from different site of collecting samples,the highest rate of infection is 69.60%in monkey farm of Nanyang,and the lowest is from Chengdu Zoo,only 4.26%.2.The study of distridution of Entamoeba species and phylogeny in nonhuman primates531 Entamoeba positive samples were selected from 1059 Entamoeba positive samples(about 50%)by random,and were used for molecular identification of Entamoeba species based on SSU rRNA,using the specific primers of E.histolytica,E.dispar,E.moshkovskii,E.nuttulli,E.coli and E.chattoni in order to understand the Entamoeba species and their distribution infecting nonhuman primates in different district,different ages and different living style.The result showed that the overall amplification efficiency was 87.19%(463/531),and only E.dispar and E.coli were amplified successfully,with 72.69%and 54.05%of amplification efficiency,respectively.The amplification efficiency of samples in Henan and Sichuan were highest,with 92.20%and 92.00%,respectively,and the lowest amplification efficiency was from samples in Guangxi,79.69%,with significant difference(x2=6.67;p<0.01).50E.dispar and 50 E.coli PCR products were collected randomly from 386 E.dispar and 287 E.coli positive PCR products to be sequenced and analyzed.The results showed that the 41 E.dispar sequences were identical to sequence(AB282661.1)of macaque-derived in Japan.The other 9 sequences have 2-3 bp difference with reference sequence(AB282661.1).The 34 E.coli sequences were identical to references sequence,and the other sequences have 3-6bp nucleotide difference.Among the 463 samples for PCR amplification,only samples from Luoyang park,Chengdu park and Kunming Zoo had single infection with E.dispar or E.coli.Thus,this study implies the possibility of infection with pathogenic E.histolytica in nonhuman primates is little,with the little risk of zoonotic transimation.3.The study of molecular epidemiology of Giardia duodenalis in dogs and catsIn order to study the infection and distribution of G.duodenalis genotypes from different sources in Zhengzhou,88 dog-derived and 5 cat-derived G.duodenalis strain were collected from April,2010 to April,2013 in Zhengzhou,and used for genotyping based on Tpi and Bg gene for G.duodenalis.The result showed that among the 93 G.duodenalis positive samples,based on Tpi gene,5 cat-derived and 54 dog-derived G.duodenalis strain were amplified successfully.The genotypes from 54 PCR positive dog samples all belonged to Assemblage C,and 5 PCR positive cat-derived strains were identified as Assemblage F.Based on Bg gene,among the 64 PCR positive dog strain,41 were Assemblage D,and 23 were Assemblage C.Cat derived samples were not amplified successfully based on Bg gene.Seven dog derived samples were infected by Assemblage C and D.4.The study of distribution of Enterocytozoon bieneusi in dogs and catsIn order to get the information of infection and distribution rule of genotypes in dogs and cats from different districts in China,276 fecal samples from dogs and cats were collected from Zhengzhou,Xinxiang,Xuchang,Xian and Chongqing in March,2012 to November,2012.They were identified using molecular method based ITS gene,and 44 targeted bands were found,with 15.94%(44/276)of total infection ratio.The infection ratio was 21.29%(34/158)and 8.47%(10/94),respectively.E.bieneusi infection ratio of stray dogs(13.89%)was lower significantly than that of pet dogs(27.91%).The infection ratio of dogs in Chongqing was 46.67%,higher than that of Zhengzhou(17.02%)and Xian dogs(11.77%).According to the result of sequence analysis,in this study 18 E.bieneusi genotype were found,of which 5 reported genotype(PtEbIX,CM1,Peru8,EbpC and Type ?)and 6 new genotype(CD1-6)from dog and 4 reported genotype(PtEbIX,BEB6,D and I)and 3 new genotype(CC1-3)were identified.Phylogenetic analysis declared that 5 new genotypes(CD1,CD2,CD4,CC2 and CC3),EbpC?D?Type ??Peru8 and CM1 belong to Group1.Group 1 is zoonotic pathogenic group,so these 5 new genotypes had zoonotic characterization,with important public health significance.Conclusion:1.The Entamoeba spp.infection ratio was 39.40%in different district of China,implying that Entamoeba spp.were the common parasites in nonhuman primates;2.Based on molecular identification of SSU rRNA,only E.dispar and E.coli were found,and the sequence of E.dispar and E.coli strains from different district had sequence disparity,indicating that pathogenic E.histolytica in nonhuman primates had little risk of zoonotic transmission,but care should be taken when suspected E.histolytica cysts were found in fecal samples of nonhuman primates by microscopy.3.G.duodenalis positive strains were found in fecal samples of cats and dogs from different sources in Zhengzhou.Based on Tpi and Bg gene identification,only Assemblage C and D were found in dog samples respectively,and Assemblage F was confirmed in cat samples,and meanwhile mixed infection with Assemblage C and D were found in samples of dogs.Perhaps dog and cat have the little impact on zoonotic transmission of G.duodenalis,but there is still the possibility of G.duodenalis transmission when contacting with dog and cat.4.The infection ratio of E.bieneusi of dog and cat in different district of China was 15.94%,and according to the sequence analysis,18 E.bieneusi genotypes were found,of which host specific genotypes are the predominant genotypes,but 10 were zoonotic genotypes,thus indicating dogs and cats have the potential risk of during the course of E.bieneusi zoonotic transmission. |
PDF Full Text Request