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Molecular detection and genetic diversity of Dientamoeba fragilis and enterocytozoon bieneusi in fecal samples submitted for routine parasitological examination

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Asian Pacific Journal of Tropical Medicine 2021年第2期14卷 73-77页

作者： ThaináValente Bertozzo érica Boarato David Ana Paula Oliveira-Arbex Semíramis Guimarães Department of Parasitology Institute of BioscienceSão Paulo State University(UNESP)BotucatuSPBrazil Tropical Diseases Postgraduate Program Medical SchoolSão Paulo State University(UNESP)BotucatuSPBrazil Sagrado Coração University(USC) Department of Health SciencesBauruSPBrazil Integrated Faculties of Bauru(FIB) BauruSPBrazil

Objective:To detect Dientamoeba(D.)fragilis and enterocytozoon(E.)bieneusi and to assess their genetic characteristics in stool samples submitted for routine examination in a clinical laboratory in Southeastern ***:In this survey,348 stool samples from female and male individuals with age ranging from 0 to≥55 years were analyzed by PCR amplifying and sequencing based on the small subunit ribosomal RNA(SSU r RNA)gene of *** and the internal transcribed spacer of ***:*** and *** isolates were observed in 2.29%(8/348)and 4.59%(16/348)of the samples,*** parasites were detected in stool samples from individuals of both genders,including young children under nine until adults over 55 years *** statistically significant differences were *** *** isolates were classified as genotype 1 and *** isolates included genotypes D(n=15)and A(n=1).Conclusions:The findings provide relevant findings on occurrence and genetic diversity of *** and ***,pointing to the need for the diagnosis of these parasites in routine examinations in clinical *** addition to sensitive diagnostic methods,it is mandatory that these parasites be considered relevant for physicians and laboratory staff.

关键词： Dientamoeba fragilis enterocytozoon bieneusi Clinical laboratory PCR Genotyping

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First report of enterocytozoon bieneusi from giant pandas(Ailuropoda melanoleuca) and red pandas(Ailurus fulgens) in China

First report of Enterocytozoon bieneusi from giant pandas(Ai...

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中国畜牧兽医学会兽医寄生虫学分会第十三次学术研讨会

作者： Tian Geru Du Shuaizhi Song Junke Lin Qing Zhao Guanghui Yu Sanke College of Veterinary Medicine Northwest A&F University

Giant pandas(Ailuropoda melanoleuca) and red pandas(Ailurus fulgens),two precious and endangered animals for wildlife conservation(Yonzon and Hunter,1991;Peng et al.,2001),are respectively classified as Category Ⅰ and Ⅱ protected species in *** infections have been identified as the most serious threat for survival of wild pandas in China currently(He et al.,2012).enterocytozoon bieneusi is an emerging and opportunistic enteric pathogen triggering diarrhea and enteric disease in humans and *** has been detected in immune-competent individuals,resulting in self-limiting diarrhea(Nkinin et al.,2007).More importantly,it is considered as an opportunistic pathogen which can cause life-threatening diarrhea in immune-compromised individuals especially in AIDS patients,raising public health concerns(Li et al.,2015).Although extensive research on this pathogen,the prevalence and genotypes of *** infection in precious wild animals of giant and red pandas have not been *** the present study,82 faecal specimens were collected from 46 giant pandas and 36 red pandas from September 2013 to June 2014 in Rare Wildlife Rescue Breeding Research Center and Xi’an Qinling Wildlife Park in Shaanxi province,northwestern *** extracting DNA,90bp fragment of the partial ribosomal internal transcribed spacer(ITS) region of *** was amplified and *** overall infection rate of 10.98%(9/82)was observed in pandas,with 8.70%(4/46) for giant pandas,and 13.89%(5/36) for red *** ITS genotypes were identified:the novel genotype I-like(n=4) and genotype EbpC(n=5).The novel genotype I-like had five nucleotide polymorphisms(transversions:T/A;C/G;T/G;C/G;transition:A/G) with the genotype I(KJ668738),with the identity of 99%.Nine positive specimens were further characterized by Multilocus sequence typing(MLST) analyses using three microsatellites(MSI,MS3 and MS7) and one minisatellite(MS4).The result showed that nine,six,six and nine positive pr

关键词： Ailuropoda melanoleuca Ailurus fulgens enterocytozoon bieneusi Prevalence Genotype

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Molecular characterization of Cryptosporidium spp.,Giardia duodenalis,and enterocytozoon bieneusi in captive wildlife at Zhengzhou Zoo,China

Molecular characterization of Cryptosporidium spp.,Giardia d...

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中国畜牧兽医学会兽医寄生虫学分会第十三次学术研讨会

作者： Junqiang Li Meng Qi Yankai Chang Rongjun Wang Tongyi Li Haiju Dong Longxian Zhang College of Animal Science and Veterinary Medicine Henan Agricultural University International Joint Research Laboratory for Zoonotic Diseases of Henan Zhengzhou Zoo

Background Cryptosporidium spp.,Giardia duodenalis,and enterocytozoon bieneusi are common gastrointestinal protists in humans and *** information is available about these parasites in captive wildlife in central *** Two hundred three fecal specimens from 80 wildlife species were collected and their genomic DNA *** were characterized with a DNA sequence analysis of the small subunit rRNA(SSU rRNA),the actin,and the 70-kDa heat shock protein(hsp70) *** duodenalis was identified by analyzing the sequences of the triosephosphate isomerase(tpi),glutamate dehydrogenase(gdh),and p-giardin(bg) *** bieneusi was genotyped with a sequence analysis of the internal transcribed spacer(ITS) region of the SSU rRNA *** Cryptosporidium felis,***,and avian genotype Ⅲ were identified in three animals(1.5%),the manul,red-crowned crane,and *** duodenalis was found in five animals(2.5%):assemblage B in a white-cheeked gibbon and beaver,and assemblage F in a Chinese leopard and Siberian *** genotypes of ***(seven previously reported genotypes and six new genotypes) were detected in 32animals(15.8%) A phylogenetic analysis of *** showed that five genotypes(three known and two new) clustered in group 1;three known genotypes clustered in group 2;one known genotype clustered in group 4;and the remaining four genotypes clustered in a new *** Zoonotic Cryptosporidium spp.,***,and *** are maintained in wildlife and transmitted between *** disease outbreaks of these infectious agents possibly originate in wildlife reservoirs.

关键词： Cryptosporidium Giardia duodenalis enterocytozoon bieneusi PCR Captive wildlife

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Occurrence of Cryptosporidium spp.,Giardia intestinalis and enterocytozoon bieneusi in captive non-human primates in Qinling Mountains

Occurrence of Cryptosporidium spp.,Giardia intestinalis and ...

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中国畜牧兽医学会兽医寄生虫学分会第十三次学术研讨会

作者： Yu Sanke Song Junke Lin Qing Zhao Guanghui College of Veterinary Medicine Northwest A&F University

Cryptosporidiosis,giardiasis and microsporidiosis,three emerging infectious diseases caused by Cryptosporidium,Giardia and microsporidia in humans and animals,have been reported both in developed countries and in those under development in last decades(Xiao L and Fayer R,2008;Thellier M and Breton J,2008).Clinical signs of these diseases are normally chronic diarrhoea in immune-competent hosts,but these infections may lead to significant morbidity and mortality in immune-compromised or-suppressed hosts,such as AIDS patients(Ryan U and Caccio SM,2013).Data from several studies suggested that non-human primates(NHPs)might be sources of cryptosporidiosis,giardiasis and microsporidiosis for human infections,since they have close relationship with human and can harbor zoonotic species/genotypes/subtypes(Nizeyi JB et al.,2002).In the present study,197 fresh faecal samples were collected from eight NHP species in Qinling Mountains,northwestern *** extracting DNA,the gene loci of triose phosphate isomerase(tpi),beta giardin(bg)and glutamate dehydrogenase(gdh) loci were used to determine *** *** 18 S rRNA gene and 60 kDa glycoprotein(gp60) was employed for the detection of Cryptosporidium species/genotypes.A 390 bp fragment of the internal transcribed spacer(ITS) gene was amplified by nested PCR to genotype *** *** results showed that 35(17.8%) samples were positive for tested parasites,including Cryptosporidium spp.(3.0%) and ***(2.0%) and ***(12.7%).*** assemblage E was detected in three *** and one Saimiri ***-variations were observed at the triose phosphate isomerase(tpi),beta giardin(bg) and glutamate dehydrogenase(gdh) loci,with three,one and two new subtypes found in respective *** *** detected in six faecal samples of Macaca mulatto,and identified as ***(n=1) and ***(n=5).*** were found in Cercopithecus neglectus(25.0%),Papio ham

关键词： Cryptosporidium spp. Giardia intestinalis enterocytozoon bieneusi Captive non-human primates Qinling Mountains

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Genotyping of enterocytozoon bieneusi in sheep and cattle in northeast China and zoonotic importance

Genotyping of Enterocytozoon bieneusi in sheep and cattle in...

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中国畜牧兽医学会兽医寄生虫学分会第十三次学术研讨会

作者：姜艳雪 Wei Tao Qiang Wan Qiao Li Yuqi Yang Yongchao Lin Siwen Zhang 李巍 College of Veterinary Medicine Northeast Agricultural University

This study investigated fecal specimens from 489 sheep and 537 cattle in multiple cities in northeast China for the prevalence and genetic characteristics of enterocytozoon bieneusi by PCR and sequencing of the ribosomal internal transcribed ***-eight sheep specimens(13.9%) and 32 cattle specimens(6.0%) were positive for *** polymorphisms enabled the identification of 9known genotypes(BEB4,BEB6,CM7,CS-4,EbpC,G,I,J,and OEB1) and 11 new genotypes(NESH1to NESH6 and NECA1 to NECA5).The genotypes formed two genetic clusters in a phylogenetic analysis,with CS-4,EbpC,G,NESH1 to NESH3,and NECA1 to NECA5 distributed in zoonotic group 1 and BEB4,BEB6,CM7,I,J,OEB1,and NESH4 to NESH6 distributed in potentially host-adapted group *** 70%of cases of *** infections in sheep were contributed by human-pathogenic genotypes BEB6,CS-4,and EbpC,and over 80%of those in cattle were by genotypes BEB4,CS-4,EbpC,Ⅰ,and *** co-occurrence of genotypes BEB4,CS-4,EbpC,I,and J in domestic ruminants and children in northeast China and the identification of BEB6 and EbpC in humans and water in central China imply the possibility of zoonotic *** study also summarizes *** genotypes obtained from ruminants worldwide and displays their host ranges,geographical distributions,and phylogenetic *** data suggest a host range expansion in some group 2 genotypes(notably BEB4,BEB6,I,and J) that were previously considered to be adapted to *** should be concerned about the increasing zoonotic importance of group 2 genotypes with low host specificity.

关键词： enterocytozoon bieneusi Sheep Cattle Genotyping Zoonosis Host-Range Expansion

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Genotypes of enterocytozoon bieneusi in dogs and cats in eastern China

Genotypes of Enterocytozoon bieneusi in dogs and cats in eas...

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第十一届全国寄生虫学青年工作者学术研讨会

作者： Wen-chao LI Jie QIN Kai WANG You-fang GU College of Animal Science Anhui Science and Technology University Chuzhou City Vocation College

Objective:enterocytozoon bieneusi is a common opportunistic pathogen found in both humans and *** animals,which often live in close proximity to humans and act as a zoonotic reservoir,may play an important role in transmitting this parasite to *** present study was conducted to evaluate the prevalence,genotypic diversity and zoonotic potential of *** in dogs and cats in eastern ***:Fecal specimens from 315 dogs and 143 cats from veterinary hospitals in eastern China were examined by internal transcribed spacer(ITS)-based ***:*** was detected in 8.6% of canine and in 1.4% of feline *** total,seven genotypes of *** were identified,including four known genotypes(Pt Eb IX,Ebp C,Type IV and D) and three novel genotypes,named CHD1,CHD2 and *** dominant genotype in dogs was Pt Eb IX(59.3%;n=16/27).Phylogenetic analysis revealed that five(CHD1,Ebp C,CHD2,D and Type IV) of the seven genotypes in this study were in the so-called zoonotic group 1,whereas genotypes Pt Eb IX belonged to the dog-specific group and genotypes CHD3 were placed in group ***:Finding zoonotic genotypes of *** in dogs and cats implies that companion animals infected with *** may pose a threat to human health.

关键词： enterocytozoon bieneusi genotype dogs cats eastern China

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Genotyping of enterocytozoon bieneusi in swine and children in northeast China and assessment of the potential for zoonotic transmission

Genotyping of Enterocytozoon bieneusi in swine and children ...

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中国畜牧兽医学会兽医寄生虫学分会第十三次学术研讨会

作者：万强 Wei Tao Yanxue Jiang Yongchao Lin Siwen Zhang Yuqi Yang Qiao Li 李巍 College of Veterinary Medicine Northeast Agricultural University

This study analyzed 893 fecal specimens of diarrheic and nondiarrheic pigs from 17 cities,northeast China for the prevalence and genetic characteristics of enterocytozoon *** parasite was detected in 422 of 893(47.3%) pigs by nested PCR of the ribosomal internal transcribed spacer(ITS),giving a prevalence of 35.1%in diarrheic animals(61/174) and 50.2%in nondiarrheic animals(361/719).The difference in prevalence between preweaned(50.4%,141/280) and growing pigs(37.5%,141/376) and between weaned(59.1%,140/237) and growing pigs is significant(p < 0.05).Genotypic typing and phylogenetic analysis facilitated identification of 15 known genotypes CC-1,CHN7,D,EbpA,EbpB,EbpC,EbpD,G,H,Henan-Ⅰ,Henan-Ⅲ,Henan-Ⅳ,O,PigEBITS3,and PigEBITS5 and 10 novel genotypes CS-1 to *** those,CS-4,D,EbpA,EbpC,EbpD,H,Henan-Ⅰ,Henan-Ⅲ,Henan-Ⅳ,O,and PigEBITS5 are human-pathogenic genotypes and CS-8,D,EbpA,EbpB,EbpC,EbpD,G,Henan-Ⅰ,and O are the contributors to water *** study also investigated fecal specimens from children of various age groups and clinic presentations for the *** prevalence(7.5%,19/255) and genotypes(CS-4,EbpC,Henan-Ⅳ,and NEC1 to NEC5).The most dominant genotype EbpC found in pigs from Changchun,Songyuan,Daqing,and Qiqihar contributed significantly to human infections in Daqing and *** CS-4 and EbpC from Harbin and Henan-Ⅳ from Qiqihar,Suihua,and Changchun determined in pigs herein represented the main causative agents of human microsporidiosis in *** study provided robust evidence that pigs could be an outstanding source of human microsporidiosis and water contamination in China.

关键词： enterocytozoon bieneusi Swine Children Genotyping Zoonosis

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Zoonotic enterocytozoon bieneusi genotypes in Pere David's deer(Elaphurus davidianus) in Henan,China

Zoonotic Enterocytozoon bieneusi genotypes in Pere David's d...

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中国畜牧兽医学会兽医寄生虫学分会第十三次学术研讨会

作者：张振杰 HUANG Jian-ying Md Robiul Karim ZHAO Jin-feng DONG Hai-ju AI Wei-chang LI Fu-huang ZHANG Long-xian 王荣军 College of Animal Science and Veterinary Medicine Henan Agricultural University The Wild Animals Rescue Center of Henan Province College of Veterinary Medicine China Agricultural University

enterocytozoon bieneusi is a zoonotic pathogen of the phylum Microspore that infects humans as well as a variety of animal species *** molecular epidemiologic studies have characterized this parasite in various hosts,isolates from many susceptible hosts have not yet been *** experiment was conducted to study enterocytozoon bieneusi genotypes in Pere David’s deer(Elaphurus davidianus) in Henan,*** this study,*** was isolated from 47 Pere David’s deer(Elaphurus davidianus) in Henan,China and characterized via PCR analysis of the internal transcribed spacer(ITS) *** was detected in 16 out of 47(34.0%) fecal specimens *** analysis of the ITS revealed six known genoty pes:Type Ⅳ(4),EbpC(4),EbpA(4),BEB6(2),COS-Ⅰ(1),and COS-Ⅱ(1).Of these,TypeⅣ,EbpC,and EbpA are known to cause human microsporidiosis worldwide,whereas the remaining genotypes are generally specific to *** present study indicated that Pere David’s deer are naturally infected with ***,predominantly with zoonotic genotypes,and may pose a risk for human transmission.

关键词： enterocytozoon bieneusi genotyping ITS Pere David’s deer.

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Prevalence and genetic characteristics of Cryptosporidium,enterocytozoon bieneusi and Giardia duodenalis in cats and dogs in Heilongjiang province,China

Prevalence and genetic characteristics of Cryptosporidium,En...

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中国畜牧兽医学会兽医寄生虫学分会第十三次学术研讨会

作者：陶威 Yanxue Jiang Qiang Wan Siwen Zhang Yuqi Yang Qiao Li 李巍 College of Veterinary Medicine Northeast Agricultural University

This study investigated 319 fecal specimens of cats(n=52) and dogs(n=267) from Heilongjiang province,China for the prevalence and genetic characteristics of Cryptosporidium,enterocytozoon bieneusi,and Giardia *** and DNA sequence analysis of the small subunit rRNA gene identified *** and *** in one cat each(3.8%) and *** and *** in 6 dogs(2.2%).Polymorphisms in the ribosomal internal transcribed spacer and phylogenetic analysis characterized zoonotic *** genotypes D,EbpC,NED1,and NED2 and host-adapted ones NED3,NED4,and PtEb IX in 18 dogs(6.7%) and human-pathogenic genotypes D and IV in 3 cats(5.8%).Genotyping based on the hypermutation of G duodenalis triosephosphate isomerase gene(TPI) facilitated identification of assemblage F in a cat(1.9%)and assemblages C and E in 12 dogs(4.5%).Subtypes of G duodenalis isolates were determined by measuring the diversity of both TPI nucleotide and amino ***,***,***,*** genotypes D,EbpC,and IV,and *** assemblage C identified herein have been documented in human infections in ***,***,***,and *** genotypes D,EbpC,and IV carried by cats or dogs also existed in wastewater in *** finding suggested pet animals could be reservoirs for human cryptosporidiosis,microsporidiosis,and giardiasis and potential sources of water contamination in China.

关键词： Cryptosporidium enterocytozoon bieneusi Giardia duodenalis Genotyping Subtyping

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Genetic characteristics of enterocytozoon bieneusi in fur animals and assessment of the potential for zoonotic transmission

Genetic characteristics of Enterocytozoon bieneusi in fur an...

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中国畜牧兽医学会兽医寄生虫学分会第一届青年科学家学术论坛

作者： Zhu Weining Yang Yuqi Tao Wei Jiang Yanxue Wan Qiang Li Qiao Li Wei Heilongjiang Key Laboratory for Zoonosis Northeast Agricultural University

various ages by sequence analysis of the ITS *** CHN-DC1 and mixed CHN-DC1/WildBoar3 were detected in one adult raccoon dog *** further explored the genotypic and phylogenetic characteristics of four mini-and microsatellites and performed a population genetic analysis of 39 *** isolates of potentially zoonotic ITS genotype D from farmed foxes and raccoon dogs in *** polymorphisms facilitated determination of six,two,four,and five genotypes at markers MSI,MS3,MS4,and MS7,*** of phylogeny revealed different levels of diversity within and among the genetic *** genotypic and phylogenetic divergences between *** isolates of ITS genotype D from fur animals and humans were observed at individual *** linkage disequilibrium and very limited recombination in subsequent population genetic analysis supported a clonal structure for *** population from for animals(FID).Phylogenetic analysis,genetic network,and measures of F and gene flow demonstrated population differentiation of FID from two known human *** populations HID(with a clonal structure) and HIA(with an epidemic structure).The data indicated an ideal resolving power of MLST compared to the previously widely used ITS genotyping and confirmed the clonal nature and population differentiation of *** in various hosts.

关键词： enterocytozoon bieneusi genotyping zoonotic transmission potential multilocus sequence typing population genetic structure host specificity

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