Genetic diversity in Echinococcus shiquicus from the plateau pika (Ochotona curzoniae) in Darlag County, Qinghai, China.
The metacestode of Echinococcus shiquicus has been recorded previously in the lung and liver of its intermediate host, the plateau pika (Ochotona curzoniae), but there is limited information regarding other organ sites. There is also limited evidence of intra-specific genetic variation within E. shiquicus. A PCR-amplified mitochondrial (mt) nad1 gene fragment (approximately 1400bp in size), with unique EcoRI and SspI restriction sites, was used to distinguish cysts or cyst-like lesions of E. shiquicus from E. multilocularis. Then, the complete mt nad1 and cox1 genes for the E. shiquicus isolates were amplified and sequenced. Phylogenetic tree and haplotype network analyses for the isolates were then generated based on a concatenated dataset of the nad1 and cox1 genes using the neighbour-joining (NJ) method and TCS1.21 software. Nineteen of eighty trapped pikas were found to harbor cysts (71 in total) when dissected at the survey site. Seventeen animals had cysts (fertile) present only in the lungs, one animal had fertile cysts in the lungs and spleen, and one individual had an infertile kidney cyst. Restriction endonuclease analysis of a fragment of the nad1 gene indicated all the cysts were due to E. shiquicus. Genetic diversity analysis revealed that the nad1 and cox1 genes varied by 0.1-1.2% and 0.1-1.0%, respectively. Haplotype network analysis of the concatenated nad1 and cox1 sequences of the isolates showed they were classified into at least 6 haplotypes, and different haplotype percentages ranged from 4.2% to 29.6%. Although, high haplotype diversity was evident in the study area, the complete nad1 and cox1 gene sequences obtained indicated that all samples represented isolates of E. shiquicus. The study has also provided a new PCR-restriction endonuclease-based method to rapidly distinguish E. shiquicus from E. multilocularis which provides a useful tool for epidemiological investigations where the two species overlap.
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Key Laboratory of Veterinary Public Health of Agriculture Ministry, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, PR China; Institute of Biomechanics and Medical Engineering, School of Aerospace, Tsinghua University, Beijing 100084, PR China.
Yan-Lei
YL
Fan
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Key Laboratory of Veterinary Public Health of Agriculture Ministry, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, PR China.
Zhong-Zi
ZZ
Lou
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Key Laboratory of Veterinary Public Health of Agriculture Ministry, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, PR China.
Li
L
Li
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Key Laboratory of Veterinary Public Health of Agriculture Ministry, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, PR China.
Hong-Bin
HB
Yan
Gansu Provincial Center for Animal Disease Control and Prevention, Lanzhou 730046, Gansu Province, PR China.
Quan-Yuan
QY
Liu
Gansu Provincial Center for Animal Disease Control and Prevention, Lanzhou 730046, Gansu Province, PR China.
Fang
F
Zhan
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Key Laboratory of Veterinary Public Health of Agriculture Ministry, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, PR China.
Jian-Qiu
JQ
Li
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Key Laboratory of Veterinary Public Health of Agriculture Ministry, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, PR China.
Cong-Nuan
CN
Liu
Laboratory of Plateau Veterinary Parasitology, Veterinary Research Institute, Qinghai Academy of Animal Science and Veterinary Medicine, Xining 810016, Qinghai Province, PR China.
Jin-Zhong
JZ
Cai
Laboratory of Plateau Veterinary Parasitology, Veterinary Research Institute, Qinghai Academy of Animal Science and Veterinary Medicine, Xining 810016, Qinghai Province, PR China.
Meng-Tong
MT
Lei
Gansu Provincial Center for Animal Disease Control and Prevention, Lanzhou 730046, Gansu Province, PR China.
Wan-Gui
WG
Shi
Molecular Parasitology Laboratory, Infectious Diseases Division, Queensland Institute of Medical Research, Brisbane, QLD 4006, Australia. Electronic address: yangyurong@hotmail.com.
Yu-Rong
YR
Yang
Molecular Parasitology Laboratory, Infectious Diseases Division, Queensland Institute of Medical Research, Brisbane, QLD 4006, Australia. Electronic address: don.mcmanus@qimrberghofer.edu.au.
Donald P
DP
McManus
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Key Laboratory of Veterinary Public Health of Agriculture Ministry, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, PR China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease, Yangzhou 225009, Jiangsu Province, PR China. Electronic address: jiawanzhong@caas.cn.
Wan-Zhong
WZ
Jia
D000818:Animals; D002681:China; D003560:Cysts / Q000469:parasitology / Q000473:pathology; D004443:Echinococcosis / Q000469:parasitology / Q000473:pathology; D004446:Echinococcus / Q000235:genetics*; D014644:Genetic Variation / Q000235:genetics*; D006239:Haplotypes / Q000235:genetics; D007793:Lagomorpha / Q000469:parasitology*; D008168:Lung / Q000469:parasitology / Q000473:pathology; D010802:Phylogeny
E. multilocularis; E. shiquicus; Genetic diversity; Haplotype analysis; Plateau pika; Restriction endonuclease analysis