Table of Content

30 October 2014, Volume 32 Issue 5

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Molecular Cloning and Localization of Leishmania donovani Expression Site Associated Genes-like Protein

LIU Peng，ZHANG Ren-gang，ZHANG Jie，JING Bao-qian

2014, 32(5):  1-327-333. 

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Objective  To clone the novel gene that specifically expressed in the amastigotes of Leishmania donovani， and observe subcellular localization of the gene encoding protein.  Methods  mRNA from promastigotes and amastigotes of L. donovani were prepared. The novel expressed sequence tag of amastigotes was selected by suppression subtractive hybridization. The expression of the novel gene in different stages of L. donovani was detected by Northern hybridization and semi-quantitative RT-PCR. The subcellular localization of the novel gene encoding protein was observed.  Results  The subtractive library of the specifically expressed sequence tag of amastigotes was constructed， and a novel gene designated as expression site associated genes-like protein（ESAGLP） gene was cloned. The full length of ESAGLP cDNA was 2 258 bp. The open-reading frame encoded a polypeptide of 620 amino acid residues. ESAGLP gene expressed only in amastigotes， the encoding protein was localized in the mitochondria.  Conclusion  The ESAGLP gene is identified as a novel gene which specifically expressed in Leishmania donovani amastigotes， and its encoding protein is localized in the mitochondria.

Retrospective Analysis of Echinococcosis Surgerical Cases in Xinjiang From 2005 to 2013

Yisilayin OSMAN，HOU Yan-yan，ZHAO Jiang-shan，Yalikun MAMTIMIN

2014, 32(5):  2-334-338. 

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Objective  To analyze the situation of echinococcosis surgerical cases in Xinjiang Uygur Autonomous Region from 2005 to 2013.  Methods  The surgery cases of echinococcosis in Xinjiang from 2005 to 2013 were collected， and analyzed with SPSS 17.0 and Epi Info 3.5.3 software.  Results  A total of 8 639 hydatid disease cases were reported during 2005-2013 from 94 counties （cities and districts） of 14 prefectures （municipalities） in the Region. The average number of annual operation cases were 960 cases（8 639/9 years）， and the annual incidence was 4.40/100 000 （960/21.81 million）. 82.8% （7 152/8 639） of report cases came from northern Xinjiang area with an incidence of 7.59/100 000， and 17.2% （1 487/8 639） distributed in southern Xinjiang area with an incidence of 1.58/100 000. There were 373 cases （4.4%， 373/8 639） reported in 2005， and increased to 1 434 cases （16.5% 1 434/8 639） in 2013（P＜0.05）. The cases mainly distributed in Yili Kazak Autonomous Prefecture （2 028 cases）， Tarbagatai Prefecture （1 218 cases）， Changji Hui Autonomous Prefecture （1 179 cases）， and Urumqi City （1 128 cases） of the northern Xinjiang area. There were 4 557 male （52.8%， 4 557/8 639） and 4 082 famale patients （47.3%， 4 082/8 639）（P＜0.05）. The age distribution showed a single-peak curve， and more patients concentrated in the age group of 31-40 years， accounting for 26.2% （2 265/8 639）. Among 8 639 cases， farmers accounted for 47.9% （4 134/8 639）.  Conclusion  The number of surgical cases of echinococcosis increases every year in Xinjiang since 2005， and the distribution of the disease shows a trend from the agricultural and pastoral areas to the towns.

Construction and Immunogenicity Analysis of the Attenuated Recombinant Salmonella typhimurium Strains Expressing Echinococcus granulosus Eg95 Antigen

WANG Zhi-sheng，WU Jing，LIN Yuan，LI Hong-bing，LIU Qiang，WANG Zhi-hui，LANG Duo-yong，YANG Wen*

2014, 32(5):  3-339-343. 

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Objective  To study the feasibility of using attenuated Salmonella typhimurium as carrier for oral immunization of Eg95 antigen of Echinococcus granulosus.  Methods  The recombinant plasmid pYA3341-Eg95 was constructed by inserting the Eg95 gene into expression vector pYA3341， and identified by the methods of PCR and enzyme digestion. The recombinant plasmid pYA3341-Eg95 was electro-transformed into attenuated S. typhimurium strains X3730 and X4550 one by one to construct the recombinant strain St-Eg95． The expression of recombinant Eg95 protein in the recombinant strains St-Eg95 was analyzed by Western blotting. The strains of St-Eg95 were passaged 10 times in vitro and the recombinant plasmids were extracted at one generation interval. The genetic stability of recombinant plasmids was identified by PCR. BALB/c mice were randomly divided into six groups （five mice per group） and inoculated orally with St-Eg95， 100 μl/mouse， at dosage of 1×10⁹， 1×10¹⁰， 1×10¹¹， and 1×10¹² cfu/ml， wild-type S. typhimurium strain（1×10⁷ cfu/ml）， and PBS， respectively. The survival rate was monitored daily for 30 days. Another 15 mice were divided into three groups and inoculated orally with St-Eg95（5×10¹⁰ cfu/ml）， X4550（pYA3341）（5×10¹⁰ cfu/ml）， and PBS， respectively， for 2 times， 0.5 ml/mouse/time， at biweekly intervals. On weeks 0， 2， 4， and 6 after the second immunization， sera were collected and tested for the presence of Eg95 antibody titers using commercially Eg antibody detection ELISA kit. The splenic lymphocyte proliferation was detected by MTT assay at 6 weeks after the second immunization.  Results  The constructed recombinant plasmid pYA3341-Eg95 was identified by enzyme digestion and PCR identification. The Eg95 protein（Mr 18 000） was expressed in the recombinant strains St-Eg95. After the recombinant strains St-Eg95 were passaged 10 times， the Eg95 gene（about 486 bp） was still amplified from St-Eg95. Safety results showed that mice inoculated orally with the St-Eg95 or PBS were all survival on the 30th day after immunization. However， all mice taking wild virulent S. typhimurium strain died within 4 days. The Eg95-specific antibodies examined by indirect ELISA were significantly higher in mice immunized with St-Eg95 than that of mice immunized with X4550（pYA3341） or PBS at 2 weeks after the second immunization（P<0.05）. The average Eg95-specific antibody titers reached up to the highest value of 1 ∶ 1 700 in mice immunized with St-Eg95 at 4 weeks after the second immunization. The lymphocyte proliferation test showed that the stimulation index value was significantly higher（P<0.05） in mice immunized with the St-Eg95（reached up to 1.94±0.15） than that in mice immunized with X4550 （pYA3341） or PBS at 6 weeks after the second immunization.  Conclusion  The recombinant oral attenuated S. typhimurium St-Eg95 was successfully constructed， and has a good safety and immunogenicity profile in mouse.

Genetic Polymorphism of the Gene Encoding the Apical Membrane Antigen-1 of Plasmodium falciparum

ZHOU Yin-fa，ZHANG Shan-ying*，LIN Yao-ying，YANG Fa-zhu，XIE Han-guo，XIAO Fang-zhen

2014, 32(5):  4-344-347. 

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Objective  To study the genetic diversity of apical membrane antigen-1 gene from Plasmodium falciparum (PfAMA-1).  Methods  Filter paper blood samples were collected from 23 imported P. falciparum malaria patients who returned to Fujian Province from 2006 to 2012. Nested PCR were used to amplify the PfAMA-1 gene. The amplified fragments were sequenced, and analyzed by bioinformatic software.  Results  All 23 samples were amplified a 505 bp band.  Thirty-two nucleotides were found to be variable， resulting in 18 haplotypes. Eight of these 18 halotypes were being reported here for the first time. The parasites collected from Africa showed the higher level of variability［haplotypes diversity （Hd）= 0.0985， nucleotide diversity （π）=0.0258］ as compared to the isolates from Asia （Hd=0.909， π=0.0221）. The average difference of dN-dS for all 23 PfAMA-1 sequences was 0.031±0.006. Sequence-based neutrality tests were not significant in Africa and Asia （P>0.05）. The minimum number of recombination events （Rm） was 10， and the linkage disequilibrium index（R2） evidently declined with the increase of nucleotide distance. A molecular phylogenetic tree constructed using the neighbor-joining method showed that the 23 isolates were assigned to three clades（G1， G2 and G3）. Most samples from Africa formed G1， and G3 contained most of Asian isolates. Conclusion  Plasmodium falciparum isolates from Africa show a higher genetic diversity than the isolates from Asia for PfAMA-1 gene.

Change of the Vα24 NKT Cells in Peripheral Blood of the Patients with Advanced Schistosomiasis and its Relation to the Degree of Hepatic Fibrosis

SUN Ting1，2，LI Gang1，2，CHEN Mao-jian1，2，NIE Hao1，2，LIAO Guo-xiang3，GONG Quan1，2 *

2014, 32(5):  5-348-351. 

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Objective  To investigate the change of Vα24 NKT cells number in peripheral blood and its correlation with the degree of hepatic fibrosis in patients with advanced schistosomiasis.  Methods  Thirty-two advanced schistosomiasis patients and 23 healthy persons were included in the study. The percentage of peripheral blood Vα24 NKT cells was determined by flow cytometry. The relevant indicators of liver function were detected by enzyme cycling method. Type-B ultrasound was used to examine the degree of hepatic fibrosis.  Results  Flow cytometry showed that the percentage of Vα24 NKT cells in advanced schistosomiasis patients［（0.23±0.09）%］ was significantly lower than that of healthy persons ［（1.44±0.62）%］（P<0.01）. Liver function test showed that the alanine aminotransferase（ALT） ［（44.78±33.42） U/L］， γ-glutamyl transpeptidase（γ-GT） ［（68.75±57.95） U/L］ and total bilirubin （Tbil）［（20.16±11.20） μmol/L］ in the patients were significantly higher than those of healthy persons［（18.77±14.19） U/L， （20.20±13.82） U/L， and （11.65±5.09） μmol/L］， respectively （P<0.01， P<0.01， P<0.05）. The percentage of Vα24 NKT cells was positively correlated with γ-GT （r=0.365， P<0.05）， but not significantly correlated with ALT， aspartate transaminase， direct bilirubin， alkaline phosphatase， albumin， albumin-globulin ratio （P＞0.05）. The percentage of Vα24 NKT cells in patients with gradesⅠ（5 cases）， Ⅱ（11 cases）， and Ⅲ（16 cases） fibrosis was （0.37±0.02）%， （0.28±0.04）%， （0.15±0.03）%， respectively （P<0.01）. The percentage of Vα24 NKT cells showed a significant negative correlation with the degree of liver fibrosis（r=-0.91， P<0.01）.  Conclusion  The percentage of Vα24 NKT cells in peripheral blood decreases with the aggravation of hepatic fibrosis in patients with advanced schistosomiasis.

Interaction between Toxoplasma GRA7 Dense Granule Protein and the Protein from Host Macrophages

XUE Feng1，2，HONG Cai-ling1，HUANG Min-jun1，2，SUN Lan1，2，GAN Shao-bo1，2，GU Jun-chao1，2 *

2014, 32(5):  6-352-356. 

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Objective  To identify the protein from host macrophages which interacted with GRA7 dense granule protein of Toxoplasma gondii， and reveal the relationship between protein interaction and infection process.  Methods  The recombinant GRA7 protein with N-terminal GST tag were used as a bait in in vitro GST Pull-down experiment， the proteins of THP-1 monocytic macrophage cell line were captured and identified by LC-MS/MS proteomics method. The in vivo protein interaction was verified by Co-IP experiment. The overexpression of the target host protein by pcDNA3.1（+） vector in THP-1 macrophage was further used to analyze the relationship between protein interaction and infection process.  Results  The captured THP-1 cell protein was about Mr 29 000， which was identified as human carbonic anhydrase 1（hCA1）. The significant in vivo protein-protein interaction between GRA7 and hCA1 was verified by Co-IP assay. The overexpression of hCA1 gene in THP-1 macrophage induced a higher propagation speed of T. gondii and the formation of the parasitophorous vacuole， but did not influence the number of the parasite.  Conclusion 　There is a significant protein interaction between Toxoplasma GRA7 dense granule protein and hCA1 enzyme from host macrophages， which is positively related with the propagation speed of T. gondii.

Cloning，Expression and Antigenic Analysis of Merozoite Surface Protein MSPDBL2-DBL2 Domain from Plasmodium falciparum

YANG Meng-jia1，WANG Su-rong1，ZHUGE Hong-xiang1 *，CHEN Jun-hu2

2014, 32(5):  7-357-360. 

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Objective  To clone and express the DBL domain of Plasmodium falciparum merozoite surface protein MSPDBL2（DBL2）, and investigate its antigenicity.  Methods  The DBL2 fragment was amplified by PCR and cloned into pET28a vector. The recombinant pET28a-DBL2 plasmid was transformed into E. coli BL21（DE3） and protein expression was induced by IPTG. The expressed product was purified by Ni-NTA affinity chromatography, and analyzed by SDS-PAGE and Western blotting.  Results  DBL2 gene fragment of Plasmodium falciparum merozoite surface protein MSPDBL2（950 bp） was obtained by PCR. The recombinant pET28a-DBL2 plasmid was identified by PCR， double enzyme digestion, and DNA sequencing. The recombinant DBL2 protein was expressed in an inclusion body form with Mr 340 000 after being induced with IPTG. Moreover, the purified recombinant DBL2 protein was recognized by sera from patients with falciparum malaria.  Conclusion  The recombinant pET28a-DBL2 plasmid has been constructed. The purified rDBL2 protein shows adequate antigenicity.

Iditification of Five Imported Cases of Plasmodium ovale wallikeri Infection in Zhejiang Province

ZHANG Ling-ling，RUAN Wei，CHEN Hua-liang，LU Qiao-yi，YAO Li-nong*

2014, 32(5):  8-361-365. 

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Objective  To identify and analyze Plasmodium ovale wallikeri in 5 imported malaria cases， who were detected positive by microscopy and negative by conventional PCR.  Methods  Epidemiological information and blood samples were collected from the five patients. The detection was conducted by microscopy， Rapid Diagnostic Test（RDT） and nested PCR with Plasmodium genus-specific， species-specific and Plasmodium ovale wallikeri-specific primers. The amplified products were sequenced and Blast analysis was performed on line in NCBI.  Results  The five patients returned from Africa， and all had a history of malaria. They were microscopically positive for Plasmodium sp.， and two cases showed Pan positive RDT result. All blood samples were negative for four Plasmodium spp. by conventional nested PCR， but positive by nested PCR with Plasmodium ovale wallikeri-specific primers. Blast analysis showed that the amplified sequences of the five cases had complete homology with P. ovale wallikeri clone RSH10 18S ribosomal RNA gene （Accession No. KF219561.1）.  Conclusion  The five cases which classified as positive by microscopy while negative by conventional PCR have been confirmed as Plasmodium ovale wallikeri infection by nested PCR with P. ovale wallikeri-specific primers.

Seroprevalance of Toxoplasma gondii Infection and Genotyping of the Isolates from Cancer Patients in Anhui，Eastern China

SHEN Qian1，WANG Lin2，FANG Qiang3，SHEN Ji-long1，ZHANG Lin-jie1 *

2014, 32(5):  9-366-370. 

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Objective  To investigate the seroprevalence of Toxoplasma gondii infection and identify the genotypes of T. gondii isolates from cancer patients in Anhui Province.  Methods  Three hundred and fifty-six blood samples were collected from outpatients and hospitalized cancer patients in Hefei， Anhui Province. IgG and gM antibodies specific to T gondii were determined by ELISA. The ELISA positive samples were subjected to detection of Toxoplasma DNA with PCR targeting a 529-bp repeat element of T. gondii. Genotyping of T. gondii isolates was performed using multilocus PCR-RFLP and 10 genetic markers， including 9 nuclear loci， sag1， sag2， sag3， btub， gra6， L358， pk1， c22-8， c29-2， and apico.  Results  Among 356 cancer patients， 21（5.9%） cases were found to be IgG-positive and 8（2.3%） were IgM-positive， and five of them were found to have both IgG and IgM antibodies. The total seroprevalence of Toxoplasma infection was 6.8%. Six PCR-positive samples were genotyped at 10 loci and two of them obtained all genetic markers and identified as the genotype of Chinese 1（ToxoDB#9）.  Conclusion  In this study， latent and active toxoplasmosis exist in the patients with malignant tumors， and two isolates are genotyped as the type of Chinese 1（ToxoDB#9） in Anhui， China.

Polymorphism Analysis of Plasmodium falciparum Histidine-rich Protein Ⅱ and Ⅲ

YANG Ying-chao，ZHANG Jin，WANG Guo-zhu，BO Shu-ying，ZHANG Ying，XIN Xiao-fang*

2014, 32(5):  10-373-376. 

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Objective  To analyze the polymorphism of Plasmodium falciparum histidine-rich protein（PfHRP） Ⅱ and Ⅲ.  Methods  Genomic DNA was isolated from blood samples of 20 patients infected with Plasmodium falciparum in Yunnan Province. Blood samples were tested by microscopy and RDTs. The Pfhrp2 and Pfhrp3 gene fragments were amplified by PCR and sequenced. The sequencing results were analyzed and compared using the bioinformatics software.  Results  20 patients infected with Plasmodium falciparum tested by microscopy and RDTs. PCR showed that the Pfhrp2 gene was with 389~986 bp, and Pfhrp3 gene with 329~640 bp. All PfHRPⅡ sequences started with type 1 repeat（AHHAHHVAD） and ended with the type 12 repeat （AHHAAAHHEAATH）. The number of type 7 （AHHAAD）， type 2（AHHAHHAAD） and type 6（AHHATD） within PfHRPⅡ was more than the other types of repeats， as well as type 16（AHHAAN） and type 17 （AHHDG） for PfHRPⅢ. Type 11 repeat （AHN） was not found from the PfHRPⅡ and PfHRPⅢ sequences.  Conclusion  There is an extensive diversity in Pfhrp2 and Pfhrp3 fragments in the individuals infected with P. falciparum in Yunnan. Some types of repeats are shared by PfHRPⅡ and PfHRPⅢ.

Progress on Parasiticidal Activity of Antimicrobial Peptides

LIU Ze-hua，ZHAO Jun-long*

2014, 32(5):  11-377-379,384. 

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Antimicrobial peptides are a kind of gene encoded， ribosome synthesized， small molecular polypeptides that have high efficiency， wide antibacterial spectrum， and low immunogenicity. Many studies have indicated that antimicrobial peptides can inhibit the growth of parasites or even kill them. This paper reviews the research progress on parasiticidal activity of the antimicrobial peptides in recent years， and presents the problems in the research.

Artemisinin Resistance in Plasmodium falciparum：Global Status and Basic Research

ZHAO Shao-min，WANG Man-yuan*

2014, 32(5):  12-380-384. 

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Artemisinin-resistant Plasmodium falciparum has been identified by WHO in the Greater Mekong subregion. While there is no report on artemisinin resistance in Africa and South America by now， related surveillance measures have been taken place. The genes related artemisinin-resistance has been identified and the molecular markers will be used for large-scale surveillance efforts to contain artemisinin resistance. The emergence and spread of artemisinin resistance worldwide is a present danger and needs more attention. This article reviews the progress of artemisinin-resistance malaria parasites and artemisinin-based combination therapies.

An Overview on the Physiological and Ecological Adaptation Mechanisms of the Overwinter Ticks

YU Zhi-jun，YANG Xiao-long，CHEN Jie，LIU Jing-ze*

2014, 32(5):  13-385-387,392. 

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The current paper introduces the recent research and development on the cryobiology of ticks， based on their overwinter behavior strategy and biochemical and physiological adaptation mechanisms， and provides detail information on the cold hardiness， biochemical and physiological mechanisms， the relationship between cold hardiness and diapause， which will give theoretical clues for subsequent research on the molecular regulation of cold hardiness of ticks.

Research Progress on Mitochondrial Genome Structure in the Phylum Apicomplexa

LI Xue-mei1，LI Xiao-bing2，3，HUANG Wei1 *

2014, 32(5):  14-388-392. 

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Mitochondria are ubiquitous organelles in all eukaryotic cells which are essential for a series of cellular processes and signal transduction. The phylum Apicomplexa includes series of unicellular eukaryotes and some of them are clinically or economically important parasites. Recent studies have demonstrated that apicomplexan parasites′ mitochondrial genomes exhibit remarkably diverse structures and they are ideal biological models to comprehend the evolution of mitochondrial genomes. This paper summarizes the mitochondrial genome structure of some representative apicomplexan, highlights their structure characteristics along with evolution process， and briefly describes their nuclear mitochondrial DNA and nuclear plastid DNA.

Design and Implementation of Command System for Emergency Events of Parasitic Diseases

WANG Qiang1，LI Shi-zhu1，FU Qing1，SHI Wei-jie2，SONG Wei2，JI Lin2，PENG Zhi-xiong3，LIN Qing-shuai3，HU Jiang-long3，ZHANG Li1，QIAN Ying-jun1，RUAN Yao1，LU Yan-xin1，XIAO Ning1，XU Xue-nian1，ZHOU Xiao-nong1*

2014, 32(5):  15-393-397. 

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Based on the requirement analysis and functional design of the command system for parasitic disease outbreaks， the system was constructed by workflow technique， function modules and technical architecture. The command system was a multi-platform system， could achieve multiple functions， such as monitoring and early warning of parasitic diseases， emergency video communication， emergency dispatcher， and emergency management. The system can meet the needs in emergency events of parasitic diseases， and increase preparedness level.

Development and Application of Information System for Epidemiological Investigation on Parasitic Diseases

HU Fei，LI Yi-feng，YUAN Min，LI Jian-ying，LIU Yue-min，LI Zhao-jun，LIN Dan-dan*

2014, 32(5):  16-398-401. 

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An information system for epidemiological investigation on parasitic diseases was developed. The foreground of this system was realized by using C# programming language， and the ACCESS database was used to implement the data management. The system was improved by continuous field application， and has been updated from version 1.0 to 3.0. The average treatment time， logic error rate， rate of sample loss， and post-processing error rate have now been reduced by 88.0%， 100%， 98.1%， and 100%， respectively （P＜0.01）. This system can reduce the human error and change the field investigation into effective and high-quality pattern.

Malaria Situation in Henan Province in 2013

YANG Cheng-yun，LU De-ling，ZHOU Rui-min，LIU Ying，ZHANG Hong-wei*，ZHAO Yu-ling

2014, 32(5):  17-370-372. 

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In 2013， 197 cases were reported in Henan Province and all of them were imported cases. Among them， 154（78.2%）were Plasmodium falciparum cases， 18（9.1%）were P. vivax cases， 2（1.0%）were P. malariae cases， and 23（11.7%） were P. ovale cases. The ratio of males to females was 97.5 ∶ 1. The average age was （37.8±9.9） years old. 183（92.9%） patients were returned from Africa. Most of the cases were peasants， export labours and workers. Cases were reported every month with 28 cases in May. The median interval from symptom appearing to diagnosis was 4 d， only 17 patients（8.6%）were diagnosed within 24 h. 61 cases（31.0%） were reported by provincial medical institutions. 60 cases（30.5%） were reported by county CDCs. A total of 193 cases recovered with chemotherapy and 4 cases died.

Prevalence and Molecular Characterization of Giardia lamblia Isolates from Goats in Anhui Province

GU You-fang，WANG Li-ke，LI Yang，LI Lei，CHU Xing-hai，XIN Di-wei，MA Cheng-xiang，XU Wen-han，WU Sheng-bing，WANG He-yin，LI Wen-chao*

2014, 32(5):  18-401-403. 

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Five hundred and six fresh fecal samples were collected from Lu′an， Fuyang， Suzhou， Chizhou, Wuhu， Chuzhou and Bozhou in Anhui Province， and detected firstly by direct smear microscopy. The microscopy-positive samples were amplified by nested PCR targeting the triosephosphate isomerase（TPI） and glutamate dehydrogenase（GDH） genes. The positive PCR products were sequenced in both directions. The sequences were analyzed by ClustalX 1.81 for sequence alignment and the neighbor-joining trees were constructed by Mega 5.05. Thirty-two out of 506 fecal specimens were diagnosed as Giardia-positive by microscopy with an infection rate of 6.3%. 23 and 16 of the samples were typed as assemblage E by the TPI （530 bp） and GDH （450 bp） genes， respectively. These findings indicated that there was a different distribution of subtypes of assemblage E in different areas. The zoonosis genotypes such as assemblage A or B was not found in the present study.

Seroepidemiological Survey of Toxoplasma gondii in Dogs in Urumqi

WANG Ping1，LU Gui-li1 *，FAN Yu-juan2，WANG Jie1，CHEN Fa-xi2，CHENG Jin1，XIA Jun1，SHAYILAN Kayizha1

2014, 32(5):  19-404-405. 

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From 2011 to 2012 in Urumqi， blood samples of 308 household dogs and of 110 stray dogs were collected from three pet hospitals and a stray dog shelter， respectively. Serum anti-Toxoplasma gondii IgG was detected by ELISA. The results showed that the overall seropositive rate was 31.8% （133/418）. The rate in household dogs and stray dogs was 29.9% （92/308） and 37.3% （41/110）， respectively （P>0.05）. Among 308 household dogs， the positive rate in males and females was 27.0% （41/152） and 32.7% （51/156）， respectively （P>0.05）. The seropositive rate in dogs ＜1 years old， 1-2 years old， and more than 2 years old was 27.1% （32/118）， 30.2% （29/96）， and 33.3% （31/94）， respectively （P>0.05）. The results revealed a high seroprevalence of T. gondii infection in dogs in Urumqi.

The Development of Clonorchis sinensis in Mice

LIU Ji-xin，SUN Yan-hong*，GUO Jia，YAO Shu-juan，SUN Yan，ZHANG Hao，ZHANG Chun-jing

2014, 32(5):  20-406，封三. 

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Freshwater fish were caught from Nenjiang River in Qiqihaer City， and examined for metacercariae of Clonorchis sinensis by the artificial digestion（pepsin-HCl） method. The metacercariae（35-40） were given orally into stomach to each Kunming mouse of infection group （50 mice）. The mice in control group were given the same amount of normal saline. The mice were sacrificed on the 5th， 10th， 15th， 20th， 25th， and 30th day after infection. Worms were collected， fixed and stained with carmine acetate， and observed under microscope. The egg-laying capacity of C. sinensis was observed in mice. 96%（48/50） mice were infected with metacercariae of C. sinensis. The recovery rate of adult worms on the 5th， 10th， 15th， 20th， 25th， and 30th day post-infection was 42.1%， 52.6%， 63.2%， 62.2%， 63.3%， and 63.2%， respectively. The first appearance of eggs in utero and feces was on the 15th and 20th day after infection， respectively. The branch of testis in worms was observed after 20 days of infection.