Genetic variability among Hymenolepis nana isolates from different geographical regions in China revealed by sequence analysis of three mitochondrial genes.

Hymenolepis nana is a common tapeworm that parasitizes in the small intestine of rodent animals and humans. The present study examined the sequence diversity of three mitochondrial (mt) genes namely NADH dehydrogenase subunits 5 (nad5), small subunit ribosomal RNA (rrnS), and ATPase subunit 6 (atp6) of H. nana from mice in different geographical regions of China. A part of the nad5 (pnad5), complete rrnS and atp6 genes were amplified separately from individual H. nana isolates using polymerase chain reaction (PCR) and then sequenced. The sequences of pnad5, rrnS, and atp6 were 710 bp, 704-711 bp, and 516 bp in length, respectively. The A + T contents of the sequences were 70.1-73.5% (pnad5), 70.1-71.7% (rrnS), and 76.6-77.9% (atp6). Sequence variation within H. nana was 0-1.4% for atp6, 0-1.7% for rrnS, and 0-0.7% for pnad5. The inter-specific sequence differences between H. nana and Hymenolepis diminuta were significantly higher, which was 31.6-31.7% (pnad5), 16.1-17.6% (rrnS), and 26.5-27.1% (atp6). Phylogenetic analysis based on the combined three sequences using the maximum parsimony (MP) method supported that H. nana is a species complex or "cryptic" species. These findings demonstrated clearly the usefulness of the three mtDNA sequences for population genetics and systematic studies of H. nana of human and animal health significance.

The complete mitochondrial genomes of five Eimeria species infecting domestic rabbits.

Rabbit coccidiosis caused by members of the genus Eimeria can cause enormous economic impact worldwide, but the genetics, epidemiology and biology of these parasites remain poorly understood. In the present study, we sequenced and annotated the complete mitochondrial (mt) genomes of five Eimeria species that commonly infect the domestic rabbits. The complete mt genomes of Eimeria intestinalis, Eimeria flavescens, Eimeria media, Eimeria vejdovskyi and Eimeria irresidua were 6261bp, 6258bp, 6168bp, 6254bp, 6259bp in length, respectively. All of the mt genomes consist of 3 genes for proteins (cytb, cox1, and cox3), 14 gene fragments for the large subunit (LSU) rRNA and 11 gene fragments for the small subunit (SSU) rRNA, but no transfer RNA (tRNA) genes. The gene order of the mt genomes is similar to that of Plasmodium, but distinct from Haemosporida and Theileria. Phylogenetic analyses based on full nucleotide sequences using Bayesian analysis revealed that the monophyly of the Eimeria of rabbits was strongly statistically supported with a Bayesian posterior probabilities. These data provide novel mtDNA markers for studying the population genetics and molecular epidemiology of the Eimeria species, and should have implications for the molecular diagnosis, prevention and control of coccidiosis in rabbits.

The complete mitochondrial genome sequence of Eimeria magna (Apicomplexa: Coccidia).

In the present study, we determined the complete mitochondrial DNA (mtDNA) sequence of Eimeria magna from rabbits for the first time, and compared its gene contents and genome organizations with that of seven Eimeria spp. from domestic chickens. The size of the complete mt genome sequence of E. magna is 6249 bp, which consists of 3 protein-coding genes (cytb, cox1 and cox3), 12 gene fragments for the large subunit (LSU) rRNA, and 7 gene fragments for the small subunit (SSU) rRNA, without transfer RNA genes, in accordance with that of Eimeria spp. from chickens. The putative direction of translation for three genes (cytb, cox1 and cox3) was the same as those of Eimeria species from domestic chickens. The content of A + T is 65.16% for E. magna mt genome (29.73% A, 35.43% T, 17.09 G and 17.75% C). The E. magna mt genome sequence provides novel mtDNA markers for studying the molecular epidemiology and population genetics of Eimeria spp. and has implications for the molecular diagnosis and control of rabbit coccidiosis.

Sequence variability in four mitochondrial genes among rabbit pinworm (Passalurus ambiguus) isolates from different localities in China.

Passalurus ambiguus is a common pinworm which parasitizes in the caecum and colon of rabbits. This study examined genetic variability among P. ambiguus isolated from naturally infected rabbits in four different provinces in China. The partial mitochondrial (mt) cytochrome c oxidase subunit 1 (pcox1), cytochrome b (pcytb) and NADH dehydrogenase subunits 1 and 5 (pnad1 and pnad5) were amplified separately from individual nematodes by PCR and sequenced. The results showed that pcox1, pcytb, pnad1 and pnad5 were 714, 663, 645 and 546 bp in length, respectively. The intra-specific sequence variations within P. ambiguus were 0-1.1% for pcox1, 0-1.2% for pcytb, 0-0.6% for pnad1 and 0-1.3% for pnad5, whereas inter-specific sequence differences with other members of the Oxyuridae were 16.2-17.3% for pcox1, 27.8-30.4% for pcytb, 20.2-24.0% for pnad1 and 27.1-30.3% for pnad5. Phylogenetic analyses using Bayesian inference (BI), maximum likelihood (ML) and maximum parsimony (MP) methods, based on the combined sequences of the four partial mtDNA sequences, revealed that all the P. ambiguus samples form monophyletic groups. This study demonstrated the existence of low-level intra-specific variation in cox1, cytb, nad1 and nad5 genes among P. ambiguus isolates from different geographic regions in China, and these four mtDNA sequences can be used as genetic markers for the population genetic studies of P. ambiguus, as well as the differentiation of P. ambiguus from other oxyurid nematodes.