RESUMO
The present study examined the variations in three mitochondrial (mt) DNA sequences, namely cytochrome b (cytb), cytochrome c oxidase subunit 3 (cox3) and NADH dehydrogenase subunit 5 (nad5), among Baylisascaris schroederi isolates from the Qinling subspecies of the giant panda in Shaanxi province, northwestern China. No differences in length were detected in the three mt fragments from different isolates. The intra-specific sequence variations within all B. schroederi samples were 0-2.6% for pcytb, 0-1.8% for pcox3 and 0-2.1% for pnad5, while the inter-specific sequence differences among members of the genus Baylisascaris were 8.2-15.2%, 6.2-15.9% and 8.4-16.0% for pcytb, pcox3, pnad5, respectively. A phylogenetic analysis of the combined sequences of pcytb, pcox3 and pnad 5 showed that all B. schroederi samples in the present study were located in two large clusters, with one cluster containing samples from giant pandas in Sichuan province. These findings provide basic information for further study of molecular epidemiology and control of B. schroederi infection in the Qinling subspecies of the giant panda and throughout China.
Assuntos
Ascaridoidea/genética , Genes Mitocondriais , Especiação Genética , Variação Genética , Ursidae/parasitologia , Animais , China , Citocromos b/genética , DNA de Helmintos/genética , DNA Mitocondrial/genética , Genoma Helmíntico/genética , Dados de Sequência Molecular , Análise de Sequência de DNA , Especificidade da EspécieRESUMO
The present study examined the prevalence and genotypes of Cryptosporidium andersoni in cattle in Shaanxi province, China. A total of 2071 fecal samples (847 from Qinchuan cattle and 1224 from dairy cattle) were examined for the presence of Cryptosporidium oocysts, and 70 samples (3.4%) were C. andersoni-positive and those positive samples were identified by PCR amplification of the small subunit ribosomal RNA (SSU rRNA) and the Cryptosporidium oocyst wall protein (COWP) genes. C. andersoni was the only species found in the examined cattle in this province. Fifty-seven C. andersoni isolates were characterized into 5 MLST subtypes using multilocus sequence typing analysis, including a new subtype in the native beef breed Qinchuan cattle. All of these C. andersoni isolates presented a clonal genetic structure. These findings provide new insights into the genetic structure of C. andersoni isolates in Shaanxi province and basic data of Cryptosporidium prevalence status, which in turn have implications for controlling cryptosporidiosis in this province.
Assuntos
Doenças dos Bovinos/epidemiologia , Criptosporidiose/epidemiologia , Criptosporidiose/veterinária , Cryptosporidium/genética , Proteínas de Protozoários/genética , RNA Ribossômico 18S/genética , Animais , Bovinos , Doenças dos Bovinos/parasitologia , China/epidemiologia , Criptosporidiose/genética , Cryptosporidium/classificação , Cryptosporidium/isolamento & purificação , DNA de Protozoário , Fezes/parasitologia , Genótipo , Tipagem de Sequências Multilocus , Oocistos/fisiologia , Filogenia , Prevalência , Proteínas de Protozoários/classificação , RNA Ribossômico 18S/classificaçãoRESUMO
The genetic variations in three mitochondrial DNA (mtDNA) regions, namely portion of cytochrome c oxidase subunit 1 (pcox1), NADH dehydrogenase subunit 1 (pnad1) and cytochrome b (pcytb), were examined for Dicrocoelium dendriticum samples isolated from different origins in Shaanxi Province, northwestern China. The intra-specific sequence differences within D. dendriticum samples were 0-0.52% for pcox1, 0-0.73% for pnad1 and 0-0.58% for pcytb. Phylogenetic analyses based on combined sequences of three mtDNA showed that all D. dendriticum samples were clustered together in same clade of Paragonimus westermani. But the phylogenetic trees could not reveal geographically genetic relationships of D. dendriticum isolates in this province. These findings will provide basic information for further study of molecular epidemiology and control of D. dendriticum infection in this province as well as in China.
Assuntos
DNA Mitocondrial/genética , Variação Genética , Platelmintos/genética , Animais , Sequência de Bases , China , Primers do DNA , Filogenia , Platelmintos/classificaçãoRESUMO
The nuclear ribosomal DNA (rDNA) region spanning 5.8S rDNA and the second internal transcribed spacer (ITS-2) of Baylisascaris schroederi isolated from the Qinling subspecies of giant panda in Shaanxi Province, China were amplified and sequenced. Sequence variations in the two rDNA regions within B. schroederi and among species in the family Ascarididae were examined. The lengths of B. schroederi 5.8S and ITS-2 rDNA sequences were 156 bp and 327 bp, respectively, and no nucleotide variation was found in these two rDNA regions among the 20 B. schroederi samples examined, and these ITS-2 sequences were identical to that of B. schroederi isolated from giant panda in Sichuan province, China. The inter-species differences in 5.8S and ITS-2 rDNA sequences among members of the family Ascarididae were 0-1.3% and 0-17.7%, respectively. Phylogenetic relationships among species in the Ascarididae were re-constructed by Bayesian inference (Bayes), maximum parsimony (MP), and maximum likelihood (ML) analyses, based on combined sequences of 5.8S and ITS-2 rDNA. All B. schroederi samples clustered together and sistered to B. transfuga with high posterior probabilities/bootstrap values, which further confirmed that nematodes isolated from the Qinling subspecies of giant panda in Shaanxi Province, China represent B. schroederi. Because of the large number of ambiguously aligned sequence positions (difficulty of inferring homology by positions), ITS-2 sequence alone is likely unsuitable for phylogenetic analyses at the family level, but the combined 5.8S and ITS-2 rDNA sequences provide alternative genetic markers for the identification of B. schroederi and for phylogenetic analysis of parasites in the family Ascarididae.
