ABSTRACT
Narrow-headed softshell turtles constitute a group of critically endangered freshwater turtles that belong to the family Trionychidae. Here, we determine the complete mitogenome of the Burmese narrow-headed softshell turtle Chitra vandijki. The length of the mitochondrial genome was 16,614 bp, composed of 13 protein-coding genes, 22 tRNA genes, two rRNA genes, and twelve noncoding regions. The phylogenetic analysis strongly indicated that C. vandijki is closely related to C. indica. The mitochondrial genome will contribute to the genetic research and conservation of C. vandijki in the future.
ABSTRACT
The northern snakehead (Channa argus) and blotched snakehead (Channa maculata) and their reciprocal hybrids have played important roles in the Chinese freshwater aquaculture industry, with an annual production in China exceeding 400 thousand tons. While these are popular aquaculture breeds in China, it is not easy to identify northern snakehead, blotched snakehead, and their hybrids. Thus, a method should be developed to identify these varieties. To distinguish between the reciprocal hybrids (C. argus â × C. maculata â and C. maculata â × C. argus â), the mitochondrial genome sequences of northern snakehead and blotched snakehead and their reciprocal hybrids were compared. Following the alignment and analysis of mtDNA sequences of northern snakehead, blotched snakehead and their hybrids, two pairs of specific primers were designed based on identified differences ranging from 12S rRNA to 16S rRNA gene. The BY1 primers amplified the same bands in the blotched snakehead and the hybrid (C. maculata â × C. argus â), while producing no products in northern snakehead and the hybrid (C. argus â × C. maculata â). Amplification with WY1 yielded the opposite results. Then, 30 individuals per fish were randomized to verify the primers, and the results showed that the primers were specific for breeds, as intended. The specific primers can not only simply distinguish between two kinds of hybrids, but also rapidly identify the two parents. This study provides a method of molecular marker identification to identify reciprocal hybrids.
Subject(s)
DNA, Mitochondrial/chemistry , Fishes/genetics , Genome, Mitochondrial , Hybridization, Genetic , Animals , Base Sequence , Fishes/classification , Genetic Markers , Molecular Sequence Data , Sequence Alignment , Species SpecificityABSTRACT
The complete mitochondrial DNA of Channa argus, Channa maculata, C. maculate â × C. argus â and C. argus â × C. maculata â were sequenced to characterize and compare their mitochondrial genomes. The lengths were 16,558, 16,559, 16,558 and 16,559 bp respectively. Start codon of 13 protein-coding genes was ATG, except that COI was GTG. The control region of the mitogenome were 907, 908, 907 and 908 bp in C. argus, C. maculata and their reciprocal hybrids (C. argus â × C. maculata â and C. maculate â × C. argus â), respectively.
Subject(s)
Chimera/genetics , DNA, Mitochondrial , Fishes/genetics , Genome, Mitochondrial , Genomics , Animals , Base Composition , Crosses, Genetic , Female , Genes, Mitochondrial , Genomics/methods , Male , Open Reading Frames , Sequence Analysis, DNAABSTRACT
The planned digital textbook Human Parasitology, the first of its kind, published by People's Medical Publishing House is an important medical education effort that features the inte-gration of digital resources. This paper introduces the digital textbook construction goal, namely using paper-based textbook for five-year undergraduates as blueprint to reasonably display the digital re-sources based on the features of the discipline. The advantages of this digital approach over traditional paper-based textbook in demonstrating parasitic morphology, lifecycle, pathogenesis, diagnosis, epi-demiology and control are discussed. Valuable experiences in planning and constructing this digital textbook are to be shared.
ABSTRACT
To direct at the weak points in traditional teaching,teaching methods have been reformed in human parasitology.This article discusses practice in teaching reform of human parasitology.
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This paper discussed the characteristics,contents and implementation of quality education of medical students during basic education.
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Objective To prepare a monoclonal antibody(McAb) against Trichinella spiralis (T.s) adult and study its protective immunity. Methods BALB/c mice were immunized with soluble antigens of 3 day adult of T.s. Immnuized spleen cells of BALB/c mice were fused with myeloma cells sp2/0. The hybridoma culture supernatant was isotyped by the Ouchterlony double diffusion technique and the specificity of the McAb was determined by immunoblotting. The protective immunity of the McAb was detected after challenging the mice by the intravenous way. Results A McAb against T.s adult was obtained. The titer of culture fluid and ascetic fluid was 1∶6 400 and 1∶204 800, respectively. This McAb was identified as IgM. Western blotting results showed this McAb can be used to identify 40 000 protein of adult worm, muscle larvae and newborn larvae of T.s. No cross reactions with antigens of Ascaris suum Goeze, Taenia solium Linnaeus, Echinococcus granulosus and Schistosoma were oberserved. The number of muscle larvae was decreased by 55.63% when giving McAb to mice intravenously. Conclusions A species specific McAb against T.s adult was established and its protective immunity was identified. This McAb can be used as a powerful tool in screening Trichinella vaccine candidates.
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Objective To observe the inhibitory effect of the antibodies against midgut-protein-ingredient of Anopheles stephensi on the oocysts of Plasmodium yoelii.Methods Female An.stephensi mosquitoes raised in laboratory were dissected and the midguts were collected.Eight BALB/c mice were immunized using midgut-protein(100 ?g/mouse,4 times with an interval of 7～10day).Ten days after the last immunization,blood was taken from mice armpit artery and serum separated.The immune active antigen of the midgut protein was analyzed by Western blotting.Protein with Mr 38 000～50 000 was separated by sephadex filtering and used to immunize 12 BALB/c mice(100 ?g/mouse,4 times with interval of 7～10 days).PBS control group was established.Seven days after the last immunization,serum antibody was detected by ELISA.When the antibody titer in immunized mice reached ≥1:2 560,mice in both groups were infected by P.yoelii(about 2?107 plasmodium-infected RBC) by abdominal injection.The mosquitoes were fed on the infected mice when the number of female gametes was higher than 2 per 10 microscopical fields 3 days later.After 9 days,the mosquitoes were dissected and the amount of oocysts in midgut was counted.Results Eight protein bands were shown in midgut-protein of An.stephensi by Western blotting and the band of Mr 38 000～50 000-midgut-protein appeared clearer.The infection rate of oocysts in the experiment and control groups were 28.70%(62/216) and 51.09%(47/92) respectively(P
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Objective To obtain the recombinant protein of an antigen gene Ts88 of Trichinella spiralis and identify the characteristics of the recombinant protein. Methods Ts88 cDNA obtained by immunoscreening the cDNA library of adult T. spiralis was subcloned into the pET 28c(+) expression vector and expressed in E.coli . Mice were immunized with the fusion protein incorporated into Freund’s adjuvant and the immune sera were collected. The titers of the Ts88 immune sera and the antigenicity of the recombinant protein were detected by ELISA and Western blotting. Immuno fluorescence test was performed in order to confirm the distribution of Ts88 protein in the worm. Results The fragment of Ts88 gene was expressed successfully in E.coli and a highly purified fusion protein was obtained. Immunization with the recombinant protein in mice produced high titers of antibodies, which recognized some components of native antigens of soluble proteins from adult worm of T. spiralis. Western blotting analysis showed that Ts88 recombinant antigen was recognized by all the positive sera, such as the sera from infected or immunized rabbits, from infected swine and from patients of trichinosis. Immuno fluorescence test confirmed that Ts88 protein mainly distributed in the cuticle surface of the worm. Conclusion The Ts88 antigen gene from T. spiralis was successfully expressed. The recombinant protein presented antigenicity.
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Objective To obtain an antigenic gene of adult Trichinella spiralis. Methods cDNA library of the adult Trichinella spiralis was screened using the sera of immunized and infected rabbits. The gene sequence was analyzed by DNAstar software and GenBank database. Results Nine positive clones were identified by immunoscreening. The clone Ts87 was sequenced and a cDNA with 1 172 bp full length was obtained using 5′ RACE technique, encoding 347 amino acids. Some possible antigen epitopes were predicted. Conclusion A novel antigenic gene of Trichinella spiralis was obtained.
ABSTRACT
Plasmodium yoelii yoelii-Anopheles stephensi system was chosen as the experimental model in studying the effects of pyrimethamine on oocyst formation of the plasmodium species. The drug was given by allowing mosquitoes to feed on infected and pyrimethamine treated mice or by feeding them directly with pyrimcthamine-sugar water. The infective rate and the number of oocysts formed after drug administration were reduced, the oocysts formed being smaller and their daily growth rate slower than that of the controls. Electron microscopic and Feulgen staining studies showed that the cytoplasm of the affected oocysts contained many vacuoles, pigment aggregations and black aggre-gates (Fig. 4). No nucleus appeared in the affected oocysts, which were presumably deteriorated and became "black spores". The amount of DNA in drug-affected oocysts was scanty. No sporozoites were found in the salivary glands of these mosquitoes. It was suggested that pyrimethamine interfered with DNA synthesis of oocysts.
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Objective To determine the genotypes and distribution of MSP2 of Plasmodium falciparum isolates in Yunnan and Hainan provinces, China.Methods The central polymorphic region of MSP2 allele was amplified by the nested PCR for genotyping of P.falciparum. Results The higher degree of polymorphism of MSP2 of P.falciparum was observed in Yunnan and Hainan. Distribution and allele frequencies differed in both provinces, indicating considerable geographical heterogeneity of parasite populations. The mixed infection of different allele type and multiplicity of infection was more frequent in Hainan than in Yunnan.Conclusion There were obvious differences in the distribution and frequencies of MSP2 alleles between Yunnan and Hainan endemic areas. MSP2 is suitable to be used as a marker gene for the genotyping of P.falciparum infection.
