Occurrence and genotypes of Cryptosporidium spp., Giardia duodenalis, and Blastocystis sp. in household, shelter, breeding, and pet market dogs in Guangzhou, southern China

Abstract Cryptosporidium spp., Giardia duodenalis, and Blastocystis sp. are common intestinal protozoans that infect humans and animals worldwide. A survey that assessed the prevalence, molecular characteristics, and zoonotic potential of these pathogens was conducted on a variety of dogs in Guangzhou, southern China. A total of 651 canine stool samples from household (n = 199), shelter (n = 149), breeding (n = 237), and pet market dogs (n = 66) were collected from eight districts in Guangzhou. Cryptosporidium spp., Giardia duodenalis, and Blastocystis sp. were detected by PCR amplification of the SSU rRNA gene. Giardia duodenalis-positive specimens were further assigned into assemblages using the glutamate dehydrogenase gene. Cryptosporidium spp., G. duodenalis, and Blastocystis sp. were found in 21 (3.2%), 20 (3.1%), and 35 (5.4%) samples, respectively. The overall prevalence of shelter dogs (40.28%, 60/149) was significantly higher than that of household (3.0%, 6/199), breeding (2.1%, 5/237), and pet market dogs (7.5%, 5/66) (χ2 = 154.72, df = 3, P < 0.001). Deworming in the past 12 months had a strong protective effect on the risk of contracting parasite infections (P < 0.001). No significant differences were detected between age or sex groups (P > 0.05). Dog-specific C. canis (n = 19) and zoonotic C. parvum (n = 2) were the only two Cryptosporidium species. Sequence analysis revealed the presence of three G. duodenalis assemblages: dog-specific assemblages D (n = 14) and C (n = 5), and cat-specific F (n = 1). Zoonotic Blastocystis ST3 (n = 28) was the dominant subtype, followed by ST1 (n = 6) and ST10 (n = 1). To our knowledge, this is the first large-scale investigation on the occurrence and molecular characteristics of Blastocystis sp. in dogs in China. Our results indicated that the dogs seemed to play a negligible role as reservoirs for Cryptosporidium spp. and G. duodenalis transmission to humans, but they are potential novel suitable hosts of Blastocystis sp. A strict sentinel surveillance system of dogs should be established to minimise the zoonotic risk of spreading blastocystosis among humans and dogs.

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Detection and genetic characterization of Giardia duodenalis in pigs from large-scale farms in Xinjiang, China

Parasite ◽

10.1051/parasite/2019056 ◽

2019 ◽

Vol 26 ◽

pp. 53

Author(s):

Bo Jing ◽

Ying Zhang ◽

Chunyan Xu ◽

Dongfang Li ◽

Jinming Xing ◽

...

Keyword(s):

Large Scale ◽

Age Groups ◽

Giardia Duodenalis ◽

Rrna Gene ◽

Ssu Rrna ◽

Ssu Rrna Gene ◽

Zoonotic Risk ◽

Significant Difference ◽

Weaning Piglets ◽

Pcr Targeting

To study the presence of Giardia duodenalis in Xinjiang, northwest China, we collected 801 fecal specimens from seven large-scale pig farms and screened them using PCR targeting the SSU rRNA gene. Twenty-one (2.6%) of the specimens from five farms were G. duodenalis-positive, with a significant difference in prevalence among different farms (0–8.7%) (p < 0.01). Giardia duodenalis prevalence was highest in fattening pigs (5.4%, 7/129), followed by sows (3.2%, 7/222), post-weaning piglets (1.8%, 5/281), and pre-weaning piglets (1.2%, 2/169), but there was no significant difference in prevalence among the age groups (p > 0.05). Sequence analysis of the SSU rRNA gene revealed that the 21 G. duodenalis strains belonged to three assemblages: A (n = 2), B (n = 16), and E (n = 3). Assemblage B was the predominant assemblage and was widely distributed in all G. duodenalis-positive farms and age groups. All G. duodenalis-positive specimens were further assayed at the β-giardin (bg), glutamate dehydrogenase (gdh), and triosephosphate isomerase (tpi) genes, and two tpi, four gdh, and two bg sequences were identified. These data indicate that pigs may be a zoonotic risk and can potentially spread G. duodenalis infection from animals to humans.

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Comparative population genetic structure of two Ixodidae ticks (Ixodes ovatus and Haemaphysalis flava) in Niigata Prefecture, Japan

10.1101/862904 ◽

2019 ◽

Author(s):

Maria Angenica Fulo Regilme ◽

Megumi Sato ◽

Tsutomu Tamura ◽

Reiko Arai ◽

Marcello Otake Sato ◽

...

Keyword(s):

Genetic Structure ◽

Large Scale ◽

Southern China ◽

Isolation By Distance ◽

Mantel Test ◽

Rrna Gene ◽

Data Set ◽

Genetic Groups ◽

Niigata Prefecture ◽

Haemaphysalis Flava

AbstractIxodid tick species such as Ixodes ovatus and Haemaphysalis flava are important vector of tick-borne diseases in Japan. In this study, we used genetic structure at two mitochondrial loci (cox1, 16S rRNA gene) to infer gene flow patterns of I. ovatus and H. flava from Niigata Prefecture, Japan. Samples were collected in 29 (I. ovatus) and 17 (H. flava) sampling locations across Niigata Prefecture (12,584.18 km2). For I. ovatus, pairwise FST and analysis of molecular variance (AMOVA) analyses of cox1 sequences indicated significant among-population differentiation. This was in contrast to H. flava, for which there were few cases of low significant pairwise differentiation. A Mantel test revealed isolation by distance and there was positive spatial autocorrelation of haplotypes in I. ovatus cox1 and 16S sequences, but non-significant results were observed in H. flava in both markers. We found three genetic groups (China 1, China 2 and Japan) in the cox1 I. ovatus tree. Newly sampled I. ovatus grouped together with a published I. ovatus sequence from northern Japan and were distinct from two other I. ovatus groups that were reported from southern China. The three genetic groups in our data set suggest the potential for cryptic species among the groups. While many factors can potentially account for the observed differences in genetic structure between the two species, including population persistence and large-scale patterns of range expansion, the differences in the mobility of hosts of tick immature stages (small mammals in I. ovatus; birds in H. flava) is possibly driving the observed patterns.

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Molecular identification and subtyping of Blastocystis sp. in laboratory rats in China

Parasite ◽

10.1051/parasite/2020035 ◽

2020 ◽

Vol 27 ◽

pp. 35

Author(s):

Junqiang Li ◽

Yueyue Yuan ◽

Yuxi Jiang ◽

Wen Wang ◽

Liqin Chao ◽

...

Keyword(s):

Pcr Amplification ◽

Small Subunit ◽

Laboratory Animals ◽

Rrna Gene ◽

Sprague Dawley ◽

Laboratory Rearing ◽

Spontaneously Hypertensive ◽

Laboratory Rats ◽

Oral Transmission ◽

Blastocystis Sp

Blastocystis sp. is a ubiquitous protist that has been frequently reported in humans and animals worldwide. A total of 355 fecal samples of experimental rats were collected from four laboratory rearing facilities in China, and Blastocystis sp. was detected by PCR amplification of the partial small subunit ribosomal (SSU) rRNA gene. Twenty-nine (8.2%, 29/355) samples were positive for Blastocystis sp., with the highest infection rate (20.7%, 24/116) in rats of the Zhengzhou1, followed by that in the Zhengzhou2 (5.0%, 2/40), Shenyang (3.0%, 3/100) and Wuhan (0) rearing facilities. Among the three rat strains, Sprague–Dawley (SD) rats had higher infection rates (11.3%, 17/151) compared to Wistar rats (8.7%, 9/104) and spontaneously hypertensive (SH) rats (3.0%, 3/100). Two Blastocystis sp. subtypes (ST4 and ST7) were identified. ST4 was the predominant subtype detected in 26 samples (89.7%). A phylogenetic analysis demonstrated that the sequences of ST4 and ST7 obtained in this study were clustered with their reference subtypes. To our knowledge, this is the first report of Blastocystis sp. in experimental rats in China. Pathogen infections in laboratory animals need to be monitored due to fecal-oral transmission.

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Molecular characterization and distribution of Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi from yaks in Tibet, China

BMC Veterinary Research ◽

10.1186/s12917-019-2172-6 ◽

2019 ◽

Vol 15 (1) ◽

Cited By ~ 3

Author(s):

Yayun Wu ◽

Yankai Chang ◽

Xiangqian Zhang ◽

Yuancai Chen ◽

Dongfang Li ◽

...

Keyword(s):

Sequence Analysis ◽

Triosephosphate Isomerase ◽

Giardia Duodenalis ◽

Molecular Data ◽

Enterocytozoon Bieneusi ◽

Rrna Gene ◽

Ssu Rrna ◽

Intensive Farming ◽

Zoonotic Risk ◽

Intestinal Pathogens

Abstract Background With worldwide distribution and importance for veterinary medicine, Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi have been found in a wide variety of vertebrate hosts. At present, few available molecular data can be used to understand the features of genetic diversity of these pathogens in areas without or less intensive farming. Dominated by grazing, Tibet is a separate geographic unit in China and yaks are in frequent contact with local herdsmen and necessary for their daily life. Therefore, to investigate the distribution of these pathogens in yaks of Tibet, 577 fecal specimens were screened using nested PCR for the presence and genotypes of the three intestinal pathogens. Results The overall prevalence of Cryptosporidium spp., G. duodenalis, and E. bieneusi were 1.4% (8/577), 1.7% (10/577), and 5.0% (29/577), respectively. Cryptosporidium andersoni (n = 7) and Cryptosporidium bovis (n = 1) were detected by sequence analysis of the SSU rRNA gene. Genotyping at the SSU rRNA and triosephosphate isomerase genes suggested that all G. duodenalis positive specimens belonged to assemblage E. Sequence analysis of the internal transcribed spacer gene identified six known E. bieneusi genotypes: BEB4 (n = 11), I (n = 6), D (n = 5), J (n = 2), CHC8 (n = 1), and BEB6 (n = 1). One subtype (A5,A4,A2,A1) for C. andersoni and three multilocus genotypes for E. bieneusi were identified by multilocus sequence typing. Conclusions We report for the first time the status of three enteric pathogens infection simultaneously for grazing yaks in Tibet. Yaks in our study are likely to impose a low zoonotic risk for humans. The molecular epidemiology data add to our knowledge of the characteristics of distribution and transmission for these pathogens in Tibet and their zoonotic potential and public health significance.

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Investigation and molecular identification of Eimeria sp. sampled from captive forest musk deer

PeerJ ◽

10.7717/peerj.11751 ◽

2021 ◽

Vol 9 ◽

pp. e11751

Author(s):

Ziwei Ren ◽

Dong Yu ◽

Wei Zhao ◽

Yan Luo ◽

Jianguo Cheng ◽

...

Keyword(s):

Molecular Identification ◽

18S Rrna ◽

Pcr Amplification ◽

Plasmid Vector ◽

18S Rrna Gene ◽

Ovis Aries ◽

Molecular Characteristics ◽

Rrna Gene ◽

Musk Deer ◽

Fecal Samples

Forest musk deer (Moschus berezovskii) is an endangered, protected species in China. Intestinal coccidiosis is a significant problem for captive forest musk deer. However, there are few reports on the prevalence and molecular characteristics of Eimeria sp. in forest musk deer. We sought to investigate the prevalence of Eimeria sp. in forest musk deer in the Sichuan and Shaanxi provinces in China. We also investigated the molecular characteristics of Eimeria sp. by analyzing the 18S rRNA gene. We collected a total of 328 fecal samples from forest musk deer on seven farms throughout the Sichuan and Shaanxi provinces. We extracted this parasite’s DNA and used this as a template for nested PCR amplification. The 18S rRNA gene fragment was associated with the plasmid vector, and these products were introduced into Escherichia coli (DH5α). The cultured bacterial solution was used as a PCR reaction template for identification purposes. We collected 328 fecal samples from forest musk deer in Lixian (n = 54), Maoxian (n = 52), Ma’erkang (n = 49), Dujiangyan (n = 55), Hanyuan (n = 41), Luding (n = 36) and Weinan (n = 41). One hundred ninety-eight (60.37%) fecal samples tested positive for Eimeria sp. . In our analysis of the 18S rRNA gene we found 34 types of Eimeria sp. with a similarity of 90.5–100%. We constructed a phylogenetic tree based on the parasite’s 18S rRNA gene sequence. Our findings indicated that the Eimeria sp. that parasitized the intestinal tract of forest musk deer was closely related to Eimeria alabamensis from Bos taurus and Eimeria ahsata from Ovis aries. To the best of our knowledge, ours was the first investigation and molecular identification of Eimeria sp. sampled from captive forest musk deer in China. Our results provide epidemiological data for the monitoring and prevention of Eimeria sp. in captive forest musk deer.

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Methylobacterium salsuginis sp. nov., isolated from seawater

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY ◽

10.1099/ijs.0.64877-0 ◽

2007 ◽

Vol 57 (8) ◽

pp. 1699-1703 ◽

Cited By ~ 22

Author(s):

Xun Wang ◽

Foday Sahr ◽

Ting Xue ◽

Baolin Sun

Keyword(s):

Type Strain ◽

Novel Species ◽

Southern China ◽

Carbon Sources ◽

Pcr Amplification ◽

Rrna Gene ◽

16S Rrna Gene Sequences ◽

Phylogenetic Groups ◽

Gene Coding ◽

Methylobacterium Oryzae

Three pink-pigmented, facultatively methylotrophic strains, designated MP1, MP2 and MRT, were isolated from seawater from southern China and characterized. Analysis of their complete 16S rRNA gene sequences revealed that they constituted three separate phylogenetic groups, showing the highest levels of similarity with respect to some members of the genus Methylobacterium. PCR amplification also showed the gene coding for the α-subunit of methanol dehydrogenase (mxaF) to be present in all strains, indicating a methylotrophic metabolism. All three strains utilized d-fructose, ethanol and nutrient agar as carbon sources, but did not utilize sucrose, citrate, acetate or formaldehyde. On the basis of the phenotypic, phylogenetic and genotypic analyses, strain MRT represents a novel species, for which the name Methylobacterium salsuginis sp. nov. is proposed, with MRT (=CGMCC 1.6474T =NCCB 100140T) as the type strain. Strains MP1 and MP2 respectively represent novel strains of the species Methylobacterium oryzae and Methylobacterium lusitanum.

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Prevalence and Subtype Distribution of Blastocystis sp. in Diarrheic Pigs in Southern China

Pathogens ◽

10.3390/pathogens10091189 ◽

2021 ◽

Vol 10 (9) ◽

pp. 1189

Author(s):

Pei Wang ◽

Sen Li ◽

Yang Zou ◽

Zhao-Wei Hong ◽

Ping Wang ◽

...

Keyword(s):

Public Health ◽

Southern China ◽

Small Subunit ◽

Rrna Gene ◽

Small Subunit Rrna ◽

Pcr Assay ◽

Potential Threat ◽

Blastocystis Sp ◽

Suckling Piglets ◽

Fattening Pigs

Blastocystis sp. is a common pathogen that infects the intestines of humans and animals, causing a threat to public health. However, little information on the prevalence and subtypes of Blastocystis sp. in diarrheic pigs in China is available. Herein, 1254 fecal samples were collected from diarrheic pigs in 37 intensive pig farms in Hunan, Jiangxi, and Fujian provinces in southern China, and the prevalence and subtypes of Blastocystis sp. were investigated. Blastocystis sp. was detected by PCR assay, which amplified the small subunit rRNA (SSU rRNA) gene. Overall prevalence of Blastocystis sp. was 31.4% (394/1254), including 21.5% (66/307), 33.1% (99/299), 58.9% (56/95), and 31.3% (173/553) in suckling piglets, weaned piglets, fattening pigs, and sows, respectively. Moreover, age and region factors were significantly related to prevalence of Blastocystis sp. (p < 0.05). Four Blastocystis sp. subtypes were identified, including ST1, ST3, ST5, and ST14. The preponderant subtype was ST5 (76.9%, 303/394). To our knowledge, ST14 was firstly found in pigs in China. The human-pathogenic subtypes (ST1, ST3, ST5, and ST14) that were observed in this study indicate a potential threat to public health. These findings provided a new sight for studying the genetic structure of Blastocystis sp.

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First identification and molecular subtyping of Blastocystis sp. in zoo animals in southwestern China

Parasites & Vectors ◽

10.1186/s13071-020-04515-2 ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Lei Deng ◽

Jingxin Yao ◽

Shanyu Chen ◽

Tingmei He ◽

Yijun Chai ◽

...

Keyword(s):

Pcr Amplification ◽

Human Infection ◽

Southwestern China ◽

Rrna Gene ◽

Genetic Characteristics ◽

Zoo Animals ◽

Blastocystis Sp ◽

Animal Hosts ◽

Red Giant ◽

Subtype Distribution

Abstract Background Blastocystis sp. is an anaerobic protozoan that parasitizes many animal hosts and the human gastrointestinal tract, and its pathogenicity is controversial. Captive wildlife may be potential reservoirs for human infection with Blastocystis sp. The present study was performed to investigate the prevalence and subtype distribution of Blastocystis sp. in zoo animals in Sichuan Province, southwestern China. Methods A total of 420 fresh fecal samples were collected from 54 captive wildlife species in four zoos in southwestern China between June 2017 and September 2019. The prevalence and subtype (ST) genetic characteristics of Blastocystis sp. were determined by PCR amplification of the barcode region of the SSU rRNA gene and phylogenetic analysis. Results Overall, 15.7% (66/420) of the animal samples and 20.7% (14/54) of the species tested were shown to be infected with Blastocystis sp. The highest prevalence of Blastocystis sp. was found in Panzhihua Zoo (24.3%), which was significantly higher than that in Chengdu Zoo (6.9%), and Xichang Zoo (2.9%) (P < 0.05). There are also significant differences in the prevalence of Blastocystis sp. among different species (P < 0.05), and the highest of Blastocystis sp. prevalence was observed in white-cheeked gibbon, black great squirrel, and red giant flying squirrel (100%). Subtype analysis of Blastocystis sp. revealed nine subtypes, including six zoonotic STs (ST1-5, and ST8) and three animal-specific STs (ST10, ST14, and ST17), with ST17 as the predominant subtype (26/66) in Blastocystis sp.-positive isolates. Conclusions To our knowledge, this is the first report on the prevalence and subtype distribution of Blastocystis sp. among captive wildlife in zoos in southwestern China. This study highlights that these animals may serve as reservoirs for human Blastocystis sp. infections.

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Parasites in stray dogs from Italy: prevalence, risk factors and management concerns

Acta Parasitologica ◽

10.1515/ap-2018-0003 ◽

2018 ◽

Vol 63 (1) ◽

pp. 27-32 ◽

Cited By ~ 9

Author(s):

Claudio De Liberato ◽

Federica Berrilli ◽

Livio Odorizi ◽

Roberto Scarcella ◽

Marco Barni ◽

...

Keyword(s):

Central Italy ◽

Giardia Duodenalis ◽

Toxocara Canis ◽

Diagnostic Techniques ◽

Stray Dogs ◽

Zoonotic Risk ◽

High Prevalence ◽

Zoonotic Parasite ◽

Shelter Dogs ◽

The Moment

Abstract Parasites are among the most common pathogens recorded in dogs. High prevalence rates are usually reported in stray and shelter dogs, as these animals are less likely to be tested and treated. In this study a survey was carried out on intestinal and lung parasites of stray dogs at the moment of the admission in a shelter in Central Italy. In the period June 2014-June 2015, 262 individual faecal samples were examined. Twelve parasitic taxa were detected. Helminths were more prevalent than protozoa (61.8% and 25.6%, respectively). Ancylostomatidae showed the higher prevalence (40.5%), followed by Giardia duodenalis (21.4%), Toxocara canis (20.6%) and Trichuris vulpis (17.6%). Angiostrongylus vasorum was the most prevalent lungworm (12.6%), Crenosoma vulpis being recorded just in one dog. G. duodenalis had a high prevalence, probably because of the diagnostic techniques adopted, actually the most sensitive available for its detection. Nevertheless, the zoonotic potential of this parasite in dogs appears reduced, as all the isolates were identified as C (24.0%) and D (76.0%) dog specific assemblages. Taeniidae constituted a high potential zoonotic risk, as from the eggs it is impossible to exclude they were Echinococcus granulosus, the most relevant zoonotic parasite in Europe. The present study underline the importance of stray dogs control, aimed at preventing or minimizing parasitic spread and zoonotic transmission.

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Identification of Cryptosporidium species, Enterocytozoon bieneusi genotypes, and Giardia duodenalis assemblages in birds in Henan, China

10.21203/rs.2.19784/v1 ◽

2019 ◽

Author(s):

Haiju Dong ◽

Ru Cheng ◽

Xinmiao Li ◽

Junqiang Li ◽

Yuancai Chen ◽

...

Keyword(s):

Migratory Birds ◽

Giardia Duodenalis ◽

Pcr Amplification ◽

Small Subunit ◽

Enterocytozoon Bieneusi ◽

Rrna Gene ◽

Fecal Samples ◽

Zoonotic Pathogens ◽

Its Gene ◽

Turtle Dove

Abstract BackgroundDomesticated, wild, and migratory birds have been known to transmit diseases such as diarrhea in humans and other animals, but studies specifically on the zoonotic pathogens Cryptosporidium spp., Enterocytozoon bieneusi, and Giardia duodenalis in birds in Henan Province, China are lacking. Hence, this study sought to characterize the prevalence of these pathogens, and to identify the different species of Cryptosporidium and their phylogenetic relationships, the genotypes of E. bieneusi, and the assemblages of G. duodenalis, in birds in the province. MethodsFresh fecal samples were collected from birds in parks and pet shops in Henan, China and were screened for the presence of the pathogens using nest-PCR amplification of the small subunit ribosomal RNA (SSU rRNA) gene and the internal transcribed spacer (ITS) gene. ResultsA total of 1,005 fecal samples were collected from 32 species of birds. 21 fecal samples (2.09%) were found positive for Cryptosporidium spp., 45 (4.48%) for E. bieneusi, and 33 (3.28%) for G. duodenalis. This study identified five Cryptosporidium species: C. baileyi (10 out of 21 fecal samples, 47.62%) in crested myna (Acridotheres cristatellus), Java sparrow (Lonchura oryzivora), Chinese hwamei (Garrulax canorus), common quail (Coturnix coturnix), and Chinese grosbeak (Eophona migratoria); C. galli (5/21, 23.81%) in Chinese blackbird (Turdus mandarinus), zebra finch (Taeniopygia guttata), and white-eyes (Zosterops sp.); C. andersoni (1/21, 4.76%) in a white-eye for the first time; C. meleagridis (4/21, 19.05%) in parrots and crested myna; and C. parvum (1/21, 4.76%) in a pigeon. Two E. bieneusi genotypes: Peru6 and PtEb I were found in pigeons and European turtle dove (Streptopelia turtur). The G. duodenalis assemblage E was detected in parrots, common hill myna, crested myna, Java sparrow, white-eyes, black-throated laughingthrush, and other birds. ConclusionsOur findings indicate that the aforementioned species of birds in Henan, China could be a source of zoonotic pathogens, such as C. meleagridis, C. andersoni, C. parvum, E. bieneusi genotype Peru6, and G. duodenalis assemblage E, that cause diseases in humans.

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