This study was conducted to define the taxonomic status of Spermophilus in the plague area of Dingbian County in Shaanxi Province, China, through the two-factor variance analysis of morphological characteristics, DNA barcoding, and chromosome karyotype analysis. The Spermophilus samples collected from Dingbian and Zhengxiang Baiqi Counties exhibited significant differences in their morphological measurements. All Spermophilus samples form two distinct branches in neighbor-joining (NJ) tree. One branch included the Spermophilus samples collected from Inner Mongolia, and the other branch included samples collected from the plague foci of Shaanxi Province and the Ningxia Region. The Spermophilus samples collected from Dingbian County had a chromosome number of 2n = 38 in 84.40% of all their cells. The Spermophilus species collected from the plague area of Dingbian County was categorized as Spermophilus alashanicus (S.alashamicus). The findings reported in this study are epidemiologically significant for monitoring plague in this region of west-central China.
Animals ; China ; Cytochromes b ; analysis ; DNA Barcoding, Taxonomic ; Electron Transport Complex IV ; analysis ; Karyotype ; Plague ; microbiology ; Sciuridae ; anatomy & histology ; classification ; genetics

We report here a human case of Taenia asiatica infection which was confirmed by genetic analyses in Dali, China. A patient was found to have symptoms of taeniasis with discharge of tapeworm proglottids. By sequencing of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene, we observed nucleotide sequence identity of 99% with T. asiatica and 96% with T. saginata. Using the cytochrome b (cytb) gene, 99% identity with T. asiatica and 96% identity with T. saginata were found. Our findings suggest that taeniasis of people in Dali, China may be mainly caused by T. asiatica.
Adult ; Animals ; China ; Cytochromes b/genetics ; Electron Transport Complex IV/genetics ; Humans ; Male ; Phylogeny ; Sequence Homology, Nucleic Acid ; Taenia/classification/genetics/isolation & purification/*physiology ; Taeniasis/*parasitology

The aim of the present study is to determine the characteristics of genotype and phenotype of Echinococcus granulosus derived from wild sheep and to compare them with the strains of E. granulosus sensu stricto (sheep-dog) and E. granulosus camel strain (camel-dog) in Iran. In Khojir National Park, near Tehran, Iran, a fertile hydatid cyst was recently found in the liver of a dead wild sheep (Ovis orientalis). The number of protoscolices (n=6,000) proved enough for an experimental infection in a dog. The characteristics of large and small hooks of metacestode were statistically determined as the sensu stricto strain but not the camel strain (P=0.5). To determine E. granulosus genotype, 20 adult worms of this type were collected from the infected dog. The second internal transcribed spacer (ITS2) of the nuclear ribosomal DNA (rDNA) and cytochrome c oxidase 1 subunit (COX1) of the mitochondrial DNA were amplified from individual adult worm by PCR. Subsequently, the PCR product was sequenced by Sanger method. The lengths of ITS2 and COX1 sequences were 378 and 857 bp, respectively, for all the sequenced samples. The amplified DNA sequences from both ribosomal and mitochondrial genes were highly similar (99% and 98%, respectively) to that of the ovine strain in the GenBank database. The results of the present study indicate that the morpho-molecular features and characteristics of E. granulosus in the Iranian wild sheep are the same as those of the sheep-dog E. granulosus sensu stricto strain.
Animals ; DNA, Helminth/genetics ; DNA, Ribosomal Spacer/genetics ; Dogs ; Echinococcosis/parasitology/*veterinary ; Echinococcus granulosus/anatomy & histology/classification/genetics/*physiology ; Electron Transport Complex IV/genetics ; *Genotype ; Iran ; *Phenotype ; Phylogeny ; Polymorphism, Restriction Fragment Length ; Sheep ; Sheep Diseases/*parasitology ; Species Specificity

To investigate the effect of diosgenin (Dgn) on chondrocytes and its relation to JAK2/STAT3 signaling pathway in mice with osteoarthritis (OA).Fifteen male C57BL/6 mice were randomly divided into three groups:control group, OA group and OA+Dgn group. After 4 weeks of treatment, the histopathological changes of cartilage tissue were observed by toluidine blue staining under light microscopy and the ultrastructure of chondrocytes was observed under electron microscopy. The primarily cultured chondrocytes of OA mice were randomly divided into 4 groups:(1) OA group, (2) Dgn group, (3) Dgn+AG490 group, (4) AG490 group. The expression of p-JAK2, p-STAT3, Bax, succinate dehydrogenase (SDH) and cytochrome c oxidase (COX) were detected by Western blotting, and superoxide dismutase (SOD) was detected using colorimetric method.The morphological observation showed that the chondrocytes of OA group presented considerable pathological changes, while the chondrocytes in OA+Dgn group maintained intact membrane. Electron microscopy observation found obvious injury in cartilage tissues of OA group, while that in OA+Dgn group remained smooth. Compared with OA group, the expressions of p-JAK2 and p-STAT3 in chondrocytes of Dgn group were increased (all<0.05), and the expressions of Bax protein, SDH, COX and SOD were decreased (all<0.05). While compared with Dgn group, the expressions of p-JAK2, p-STAT3, SDH, COX and SOD in chondrocytes of Dgn+AG490 group were decreased (all<0.05), and the expression of Bax protein was increased (<0.05).Diosgenin can inhibit apoptosis and increase mitochondrial oxidative stress capacity of chondrocytes in mice with osteoarthritis, which is closely related to the activation of JAK2/STAT3 signaling pathway.
Animals ; Apoptosis ; drug effects ; Cartilage ; drug effects ; pathology ; Chondrocytes ; chemistry ; drug effects ; pathology ; Diosgenin ; pharmacology ; Electron Transport Complex IV ; metabolism ; Janus Kinase 2 ; drug effects ; Male ; Mice ; Mice, Inbred C57BL ; Mitochondria ; drug effects ; genetics ; Osteoarthritis ; genetics ; physiopathology ; Oxidative Stress ; drug effects ; STAT3 Transcription Factor ; drug effects ; Signal Transduction ; Succinate Dehydrogenase ; metabolism ; Superoxide Dismutase ; metabolism ; Tyrphostins ; pharmacology ; bcl-2-Associated X Protein ; metabolism

INTRODUCTIONAlthough there have been several phylogenetic studies on Plasmodium knowlesi (P. knowlesi), only cytochrome c oxidase subunit 1 (COX1) gene analysis has shown some geographical differentiation between the isolates of different countries.

METHODSPhylogenetic analysis of locally acquired P. knowlesi infections, based on circumsporozoite, small subunit ribosomal ribonucleic acid (SSU rRNA), merozoite surface protein 1 and COX1 gene targets, was performed. The results were compared with the published sequences of regional isolates from Malaysia and Thailand.

RESULTSPhylogenetic analysis of the circumsporozoite, SSU rRNA and merozoite surface protein 1 gene sequences for regional P. knowlesi isolates showed no obvious differentiation that could be attributed to their geographical origin. However, COX1 gene analysis showed that it was possible to differentiate between Singapore-acquired P. knowlesi infections and P. knowlesi infections from Peninsular Malaysia and Sarawak, Borneo, Malaysia.

CONCLUSIONThe ability to differentiate between locally acquired P. knowlesi infections and imported P. knowlesi infections has important utility for the monitoring of P. knowlesi malaria control programmes in Singapore.

Electron Transport Complex IV ; genetics ; Genetic Markers ; Geography ; Humans ; Malaria ; Malaysia ; Phylogeny ; Plasmodium knowlesi ; genetics ; Polymerase Chain Reaction ; Singapore ; Thailand

Human diphyllobothriasis is a widespread fish-borne zoonosis caused by the infection with broad tapeworms belonging to the genus Diphyllobothrium. In mainland China, so far 20 human cases of Diphyllobothrium infections have been reported, and the etiologic species were identified as D. latum and D. nihonkaiense based on morphological characteristics or molecular analysis. In the present study, proglottids of diphyllobothriid tapeworms from 3 human cases that occurred in Heilongjiang Province, China were identified as D. nihonkaiense by sequencing mitochondrial cytochrome c oxidase subunit I (cox1) and NADH dehydrogenase subunit 5 (nad5) genes. Two different cox1 gene sequences were obtained. One sequence showed 100% homology with those from humans in Japan. The remaining cox1 gene sequence and 2 different nad5 gene sequences obtained were not described previously, and might reflect endemic genetic characterizations. D. nihonkaiense might also be a major causative species of human diphyllobothriasis in China. Meanwhile, the finding of the first pediatric case of D. nihonkaiense infection in China suggests that infants infected with D. nihonkaiense should not be ignored.
Adult ; Animals ; China ; Diphyllobothriasis/*parasitology ; Diphyllobothrium/classification/enzymology/*genetics/*isolation & purification ; Electron Transport Complex IV/genetics ; Female ; Helminth Proteins/genetics ; Humans ; Infant

Tapeworms of the genus Spirometra are pseudophyllidean cestodes endemic in Korea. At present, it is unclear which Spirometra species are responsible for causing human infections, and little information is available on the epidemiological profiles of Spirometra species infecting humans in Korea. Between 1979 and 2009, a total of 50 spargana from human patients and 2 adult specimens obtained from experimentally infected carnivorous animals were analyzed according to genetic and taxonomic criteria and classified as Spirometra erinaceieuropaei or Spirometra decipiens depending on the morphology. Morphologically, S. erinaceieuropaei and S. decipiens are different in that the spirally coiled uterus in S. erinaceieuropaei has 5-7 complete coils, while in S. decipiens it has only 4.5 coils. In addition, there is a 9.3% (146/1,566) sequence different between S. erinaceieuropaei and S. decipiens in the cox1 gene. Partial cox1 sequences (390 bp) from 35 Korean isolates showed 99.4% (388/390) similarity with the reference sequence of S. erinaceieuropaei from Korea (G1724; GenBank KJ599680) and an additional 15 Korean isolates revealed 99.2% (387/390) similarity with the reference sequences of S. decipiens from Korea (G1657; GenBank KJ599679). Based on morphologic and molecular databases, the estimated population ratio of S. erinaceieuropaei to S. decipiens was 35: 15. Our results indicate that both S. erinaceieuropaei and S. decipiens found in Korea infect humans, with S. erinaceieuropaei being 2 times more prevalent than S. decipiens. This study is the first to report human sparganosis caused by S. decipiens in humans in Korea.
Adult ; Aged ; Animals ; Cat Diseases/parasitology ; Cats ; Dog Diseases/parasitology ; Dogs ; Electron Transport Complex IV/genetics ; Female ; Helminth Proteins/genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Phylogeny ; Republic of Korea/epidemiology ; Sparganosis/diagnosis/*parasitology ; Spirometra/anatomy & histology/classification/*genetics/*isolation & purification ; Young Adult

OBJECTIVETo study the population genetic variation, genetic diversity and phylogenesis of Anopheles sinensis in China.

METHODSAnopheles sinensis samples collected from Shandong, Anhui, Jiangsu, Guizhou, and Yunnan Provinces and Guangxi Zhuang Autonomous Region with different geographical conditions between 2010 and 2012 were analyzed by mitochondrial DNA cytochrome oxidase subunit I (mtDNA-COI) gene amplification and sequencing. Bioedit 7.0 and DnaSP 5.0 software was used to compare the gene sequences and analyze the population genetic structure, respectively. Arlequin 3.1 was used to calculate the genetic distance and parameters of population differentiation. The relationship between the geographic and genetic distances was analyzed using IBD Web Service. PHYLIP 3.6 was used to construct the phylogenetic tree.

RESULTSPCR amplification and sequencing was performed successfully for 6 Anopheles sinensis populations containing 123 female mosquitoes. The length of mtDNA-COI gene fragment was 841 bp with an average A+T content of 71.2% and G+C content of 28.8%. High nucleotide diversity and genetic differentiation were observed among the Anopheles sinensis populations based on mtDNA-COI gene. Analysis of the molecular variance revealed a greater variation between populations than that within populations with isolation by distance between the populations. The Anopheles sinensis populations appeared to have undergone expansion, but the Yunnan population constituted an isolated branch in the phylogenetic tree.

CONCLUSIONmtDNA-COI can serve as the molecular marker to analyze population genetic variation and phylogenesis of Anopheles sinensis. The Yunnan population shows a phylogenetic difference from the other populations analyzed in this study.

Animals ; Anopheles ; genetics ; China ; DNA, Mitochondrial ; genetics ; Electron Transport Complex IV ; genetics ; Female ; Genetic Variation ; Genetics, Population ; Phylogeny

Fascioliasis, a food-borne trematode zoonosis, is a disease primarily in cattle and sheep and occasionally in humans. Water dropwort (Oenanthe javanica), an aquatic perennial herb, is a common second intermediate host of Fasciola, and the fresh stems and leaves are widely used as a seasoning in the Korean diet. However, no information regarding Fasciola species contamination in water dropwort is available. Here, we collected 500 samples of water dropwort in 3 areas in Korea during February and March 2015, and the water dropwort contamination of Fasciola species was monitored by DNA sequencing analysis of the Fasciola hepatica and Fasciola gigantica specific mitochondrial cytochrome c oxidase subunit 1 (cox1) and nuclear ribosomal internal transcribed spacer 2 (ITS-2). Among the 500 samples assessed, the presence of F. hepatica cox1 and 1TS-2 markers were detected in 2 samples, and F. hepatica contamination was confirmed by sequencing analysis. The nucleotide sequences of cox1 PCR products from the 2 F. hepatica-contaminated samples were 96.5% identical to the F. hepatica cox1 sequences in GenBank, whereas F. gigantica cox1 sequences were 46.8% similar with the sequence detected from the cox1 positive samples. However, F. gigantica cox1 and ITS-2 markers were not detected by PCR in the 500 samples of water dropwort. Collectively, in this survey of the water dropwort contamination with Fasciola species, very low prevalence of F. hepatica contamination was detected in the samples.
Animals ; Base Sequence ; Cluster Analysis ; DNA, Helminth/chemistry/genetics ; DNA, Ribosomal Spacer/chemistry/*genetics ; Electron Transport Complex IV/*genetics ; Fasciola hepatica/*genetics/*isolation & purification ; Korea ; Molecular Sequence Data ; Oenanthe/*parasitology ; Phylogeny ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid

Diphyllobothrium latum and Diphyllobothrium nihonkaiense are morphologically similar to each other, and only genetic method can differentiate clearly between the 2 species. A strobila of diphyllobothriid tapeworm discharged from a 7-year-old boy was analyzed to identify the species by mitochondrial cytochrome c oxidase subunit 1 (cox1) gene sequencing. He and his family (total 4 persons) ate slices of 3 kinds of raw fish 16 days before visiting our outpatient clinic. All family members complained of abdominal pain and watery diarrhea. They all expelled tapeworm strobilae in their stools. They were treated with a single oral dose of praziquantel and then complained of no more symptoms. The cox1 gene sequencing of the strobila from the boy revealed 99.9% (687/688 bp) similarity with D. nihonkaiense and only 93.2% (641/688 bp) similarity with D. latum. Thus, we assigned this tapeworm as D. nihonkaiense. This is the first report of D. nihonkaiense infection in a family in Korea, and this report includes the 8th pediatric case in Korea. The current report is meaningful because D. nihonkaiense infection within a family is rare.
Animals ; Anthelmintics/therapeutic use ; Child ; Diphyllobothriasis/*diagnosis/drug therapy/*parasitology ; Diphyllobothrium/*classification/enzymology/genetics/*isolation & purification ; Electron Transport Complex IV/genetics ; *Family Health ; Humans ; Korea ; Male ; Praziquantel/therapeutic use ; Sequence Analysis, DNA ; Treatment Outcome