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Xiao J.-Y.,Jiamusi University | Gao J.-F.,Heilongjiang Institute of Veterinary Science | Cai L.-S.,Jiamusi University | Dai Y.,Jiamusi University | And 4 more authors.
Mitochondrial DNA | Year: 2013

The present study examined genetic variability among Clonorchis sinensis isolates from four different geographical localities (Guangzhou, Nanning, Jiamusi and Daqing) and host species (cats, dogs, human and rabbits) in Mainland China by sequence analyses of two mitochondrial DNA (mtDNA) genes, namely NADH dehydrogenase subunits 2, 5 (nad2 and nad5) and ribosomal internal transcribed spacer 1 (ITS1). A portion of the ITS1, nad2 (pnad2) and nad5 (pnad5) was amplified by polymerase chain reaction separately from adult C. sinensis individuals and the amplicons were subjected to sequencing from both directions. The length of the sequences of ITS1, pnad2 and pnad5 was 643, 666 and 771 bp, respectively. The intraspecific sequence variations within C. sinensis were 0-1.7% for ITS1, 0-1.4% for pnad2 and 0-0.9% for pnad5. The interspecific sequence variations within other zoonotic trematodes, which were published previously, were 4.5-84.9% for ITS1, 21.9-43.6% for pnad2 and 19.2-48.9% for pnad5. The A+T contents of the sequences were 45.26-45.88% (ITS1), 62.91-63.51% (pnad2) and 58.24-58.63% (pnad5). Phylogenetic analyses using ribosomal and mitochondrial sequence data set, with three different computational algorithms (Bayesian inference, maximum parsimony and maximum likelihood), all revealed distinct groups with high statistical support. These findings demonstrated the existence of low-level intraspecific variations in ribosomal DNA (rDNA) and mtDNA sequences among C. sinensis isolates from four different regions and hosts in China and elucidated that mtDNA sequences and rDNA sequences provided reliable genetic markers for phylogenetic studies of zoonotic trematodes. © 2013 Informa UK, Ltd. Source


Liu G.-H.,Chinese Academy of Agricultural Sciences | Xu M.-J.,Chinese Academy of Agricultural Sciences | Xu M.-J.,South China Agricultural University | Chang Q.-C.,Heilongjiang Bayi Agricultural University | And 4 more authors.
Parasites and Vectors | Year: 2016

Background: Schistosoma turkestanicum is a parasite of considerable veterinary importance as an agent of animal schistosomiasis in many countries, including China. The S. turkestanicum cercariae can also infect humans, causing cercarial dermatitis in many countries and regions of the world. In spite of its significance as a pathogen of animals and humans, there is little transcriptomic and genomic data in the public databases. Methods: Herein, we performed the transcriptome Illumina RNA sequencing (RNA-seq) of adult males and females of S. turkestanicum and de novo transcriptome assembly. Results: Approximately 81.1 (female) and 80.5 (male) million high-quality clean reads were obtained and then 29,526 (female) and 41,346 (male) unigenes were assembled. A total of 34,624 unigenes were produced from S. turkestanicum females and males, with an average length of 878 nucleotides (nt) and N50 of 1480 nt. Of these unigenes, 25,158 (72.7 %) were annotated by blast searches against the NCBI non-redundant protein database. Among these, 21,995 (63.5 %), 22,189 (64.1 %) and 13,754 (39.7 %) of the unigenes had significant similarity in the NCBI non-redundant protein (NR), non-redundant nucleotide (NT) and Swiss-Prot databases, respectively. In addition, 3150 unigenes were identified to be expressed specifically in females and 1014 unigenes were identified to be expressed specifically in males. Interestingly, several pathways associated with gonadal development and sex maintenance were found, including the Wnt signaling pathway (103; 2 %) and progesterone-mediated oocyte maturation (77; 1.5 %). Conclusions: The present study characterized and compared the transcriptomes of adult female and male blood fluke, S. turkestanicum. These results will not only serve as valuable resources for future functional genomics studies to understand the molecular aspects of S. turkestanicum, but also will provide essential information for ongoing whole genome sequencing efforts on this pathogenic blood fluke. © 2016 Liu et al. Source


Na L.,Chinese Academy of Agricultural Sciences | Na L.,Heilongjiang Bayi Agricultural University | Gao J.-F.,Heilongjiang Bayi Agricultural University | Gao J.-F.,Heilongjiang Institute of Veterinary Science | And 8 more authors.
Infection, Genetics and Evolution | Year: 2016

Metorchis orientalis (Trematoda: Opisthorchiidae) is an important trematode infecting many animals and humans, causing metorchiasis. In the present study, the complete mitochondrial (mt) genome of M. orientalis was sequenced. The complete mt genome of M. orientalis is 13,834. bp circular DNA molecule and contains 12 protein-coding genes, 22 transfer RNA genes, and two ribosomal RNA genes. The gene content and arrangement of M. orientalis is the same as those of Opisthorchiidae trematodes (Opisthorchis viverrini, Opisthorchis felineus and Clonorchis sinensis), but distinct from Schistosoma spp. Phylogenetic analyses using concatenated amino acid sequences of 12 protein-coding genes with three different computational algorithms (Bayesian inference, maximum likelihood and maximum parsimony) revealed that M. orientalis and O. viverrini represent sister taxa. The mt genome provides a novel genetic marker for further studies of the identification, classification and molecular epidemiology of Opisthorchiidae trematodes, and should have implications for the diagnosis, prevention and control of metorchiasis in animals and humans. © 2016 Elsevier B.V. Source


Lou Y.,Heilongjiang Bayi Agricultural University | Zhang Y.,Heilongjiang Bayi Agricultural University | Qiu J.-H.,Heilongjiang Bayi Agricultural University | Gao J.-F.,Heilongjiang Institute of Veterinary Science | And 4 more authors.
Mitochondrial DNA | Year: 2015

This research aimed at exploring sequence variability in four mitochondrial (mt) genes, namely, cytochrome c oxidase subunit 1 (cox1), cytochrome b (cytb) and NADH dehydrogenase subunits 1 and 5 (nad1 and nad5), among pinworm Aspicularis tetraptera isolates from laboratory mice in four different provinces, China. A part of the cox1 (pcox1), cytb (pcytb), nad1 and nad5 genes (pnad1 and pnad5) were amplified separately from individual pinworms by polymerase chain reaction (PCR) and sequenced to determine sequence variations and examine their phylogenetic relationships. Herein, the intra-specific sequence variations within A. tetraptera were 0-0.5% for pcox1, 0-1.4% for pcytb, 0-1.8% for pnad1 and 0-1.7% for pnad5, respectively. In contrast, the inter-specific sequence differences among members of the Oxyuridae were significantly higher, being 13.7-17.0% for pcox1, 24.5-34.7% for pcytb, 26.6-29.6% for pnad1 and 24.4-25.5% for pnad5, respectively. Three methods, namely, Bayesian inference (BI), maximum likelihood (ML) and maximum parsimony (MP), were used for phylogenetic analyses based on the combined sequences of the four mt gene sequences, and the results indicated that all A. tetraptera samples form monophyletic groups, but samples from the same geographical origin did not always cluster together. This study demonstrated the existence of low-level intra-specific variation in four mtDNA sequences among A. tetraptera isolates from laboratory mice in different geographic regions in China, indicating no obvious geographical distinction among A. tetraptera isolates in China. These findings have important implications for studying systematics, molecular epidemiology and population genetics of A. tetraptera. © 2014 Informa UK Ltd. All rights reserved: reproduction in whole or part not permitted. Source


Gao J.-F.,Heilongjiang Bayi Agricultural University | Gao J.-F.,Heilongjiang Institute of Veterinary Science | Zhao Q.,Jilin Agricultural University | Liu G.-H.,Hunan Agricultural University | And 9 more authors.
Gene | Year: 2014

Bunostomum trigonocephalum and Bunostomum phlebotomum are blood-feeding hookworms of sheep and cattle, causing considerable economic losses to the live stock industries. Studying genetic variability within and among hookworm populations is critical to addressing epidemiological and ecological questions. Mitochondrial (mt) DNA is known to provide useful markers for investigations of population genetics of hookworms, but mt genome sequence data are scant. In the present study, the complete mitochondrial DNA (mtDNA) sequences of the sheep and goat hookworm B. trigonocephalum were determined for the first time, and the mt genome of B. phlebotomum from yak in China was also sequenced for comparative analyses of their gene contents and genome organizations. The lengths of mt DNA sequences of B. trigonocephalum sheep isolate, B. trigonocephalum goat isolate and B. phlebotomum China yak isolate were 13,764. bp, 13,771. bp and 13,803. bp in size, respectively. The identity of the mt genomes was 99.7% between B. trigonocephalum sheep isolate and B. trigonocephalum goat isolate. The identity of B. phlebotomum China yak isolate mt genomes was 85.3% with B. trigonocephalum sheep isolate, and 85.2% with B. trigonocephalum goat isolate. All the mt genes of the two hookworms were transcribed in the same direction and gene arrangements were consistent with those of the GA3 type, including 12 protein-coding genes, 2 rRNA genes and 22 tRNA genes, but lacking ATP synthetase subunit 8 gene. The mt genomes of B. trigonocephalum and B. phlebotomum were similar to prefer bases A and T, the contents of A. +. T are 76.5% (sheep isolate), 76.4% (goat isolate) and 76.9% (China yak isolate), respectively. Phylogenetic relationships reconstructed using concatenated amino acid sequences of 12 protein-coding genes with three methods (maximum likelihood, Bayesian inference and neighbor joining) revealed that the B. trigonocephalum and B. phlebotomum represent distinct but closely-related species. These data provide novel and useful genetic markers for studying the systematics, and population genetics of the two ruminant hookworms. © 2014 Elsevier B.V. Source

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