Table of Content

28 February 2019, Volume 37 Issue 1

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ORIGINAL ARTICLES

Survey of common human parasite infections in Fujian Province in 2015

Bao-jian CHEN, Han-guo XIE, Rong-yan ZHANG, Chen-xin LIN, Yan-rong LI, Xian-liang XIE, Dian-wei JIAN, Shan-ying ZHANG

2019, 37(1):  1-6.  doi:10.12140/j.issn.1000-7423.2019.01.001

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Objective To investigate current endemic status of common human parasite infections in Fujian Province in order to provide scientific basis for formulating better prevention and control measures. Methods Based on the Third National Survey Plan for Common Human Parasites, the survey sites were selected by using stratified cluster random sampling method in 2015. Not less than 250 permanent residents over 3 years old were enrolled in each site. The fecal samples were collected from each participant and the helminthic eggs were examined under microscope using Kato-Katz thick smear method in duplicate. The cysts and trophozoites of intestinal protozoa were examined using direct saline smear and iodine staining. Results Total 41 survey sites were selected for soil-transmitted helminth (STH) infections, 75 sites selected for intestinal protozoa infections from 15 counties. For survey of Clonorchis sinensis infection, total 12 sites were selected from 7 cites (urban) and 10 sites selected from 5 towns (suburban). A total of 16 799 residents from 138 survey sites from 27 counties (9 prefectures) were examined, and the overall prevalence of all parasite infection was 3.69% (618/16 799). For STH infections, totally 10 652 residents were examined with overall prevalence of 3.67% (391/10 652) identified. Three STH have been identified with individual prevalence for Ascaris lumbricoides 0.06% (7/10 652), hookworms 3.36% (358/10 652) and Trichuris trichiura 0.27% (29/10 652). For intestinal protozoan infections, a total of 10 652 residents were examined and the overall prevalence was 2.08% (222/10 652), with individual prevalence for Blastocystis hominis 0.79% (85/10 652), Endolimax nana 0.58% (62/10 652), Entamoeba coli 0.41% (44/10 652), E. hartmanni 0.18% (19/10 652), E. polecki 0.09% (10/10 652), and Giardia lamblia 0.02% (2/10 652). For C. sinensis infections, total 6 147 residents were examined with over prevalence of 0.09% (6/6 147), 2 of them were identified in urban areas (0.05%, 2/3 418) and 4 identified in suburban areas (0.15%, 4/2 729）. There is no statistical difference for the prevalence of C. sinensis infection identified in urban and suburban regions (P > 0.05). For STH infection, there was no significant difference in the prevalence between male (3.31%) and females (4.01%) (P > 0.05). People with 65-years old or above had the highest prevalence of 7.87%, and people with age less than 15 years old had the lowest prevalence (0.38%), with significant difference for the distribution of STH infections in different groups of age (P < 0.01). Famers had the highest prevalence of 4.63% compared to people with other occupations (P < 0.01). People with low education (primary school or less) had the highest prevalence of STH infection (4.85%) and people with high education (university or above) had the lowest prevalence (1.29%), with significant difference compared with people with different education levels (P < 0.01). Among the 15 counties investigated, the prevalence of STH was highest in Ninghua (16.59%) and lowest in Zhouning (0). Among the 9 prefectures investigated, the highest prevalence of STH was observed in Sanming (8.87%) and lowest in Ningde (0.19%). Compared with different ecological zones, people living in Fujian/Zhejiang mountain zone had higher prevalence of STH (4.12%, 226/5 480) than people living in Yunnan/Guizhou/Guangdong/South Fujian ecological zone (3.19%, 165/5 172), with statistical difference (P < 0.05). Compared to the prevalence of STH identified at the first survey (95.65%) and the second survey (89.85%), the prevalence of STH identified at current survey has dropped 95.65% and 89.85%, respectively. Conclusion The total prevalence of human common parasite infections is greatly reduced in Fujian Province identified in this survey, and has fallen down to its lowest level in history. Among STH, hookworm infection still needs to be better controlled.

Common human parasite infections and people’s awareness survey in Tianjin in 2015

Yi-fang LIU, Jie LV, Cheng SU, Lin LI

2019, 37(1):  7-11.  doi:10.12140/j.issn.1000-7423.2019.01.002

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Objective To investigate the infection status of common parasite infections and people’s awareness of parasitic diseases and health behavior in Tianjin. Methods An epidemiological survey was carried out in Tianjin according to the requirements of the National Survey on the Status of important human parasitic diseases. The pilot survey districts were randomly selected based on the different ecologic zones and economic status. Selected districts were staratified according to the topographic, economic level and other factors. The pilot sites were selected from each layer by random number method and no less than 250 residents with age over 0 year-old was included in each site. Fecal samples were collected and the helminth eggs (soil-transmitted nematodes, tape worms and Clonorchis sinensis) were examined with the Kato-Katz method and the intestinal protozoa were examined by direct smear method. The anal tape test was used to examine Enterobius vermicularis eggs in children with 3-6 years old. The infection status among residents was analyzed based on their different gender, age and occupation with statistical analysis using Chi-square test. Some residents from 26 pilot survey sites were randomly selected to conduct a questionnaire on parasite-related knowledge to understand their awareness of parasites and infection risk. Results A total of 15 674 people from 61 pilot sites in 12 counties were investigated. The infection rate for all intestinal parasites was 0.14% （22/15 674） with only 8 infected with Ascaris lumbricoides (1 cases of moderate infection and 7 cases of mild infection) and 14 infected with Enterobius vermicularis (all of them were mild infection). The infection rate of E. vermicularis in children with age 3-6 was 0.54% (4/744). There was no other intestinal parasite identified. The infection rate for A. lumbricoides in Yanshan-Taihang Mountain Ecological Zone was 0.17% (8/4 705) and no parasite infection was detected in Beijing-Tianjin-Tangshan Ecological Zone. All infections happened in Wuqing, Binhai, Jinghai, Baodi and Ji Districts (all are agriculture districts) with parasite infection rates of 0.13% (1/784), 0.07% (1/1 501), 0.29% (3/1 040), 1.04% (8/766) and 0.19% (9/4 705), respectively. The difference was statistically significant among counties (P < 0.05). The parasite infection rates in males and females were 0.09% (7/7 566) and 0.19% (15/8 108). The infection happened in farmers (0.18%, 14/7 620) was higher than those with other occupations. The infection rate in age of 50-59 (0.23%, 6/2 658) was higher than other age groups. Surprisingly, people with high school education or above had higher infection rate (0.21%, 7/3 330) than other education groups, however, the infection rate showed no significant difference among genders, age groups, levels of education and occupations (P > 0.05). Totally 1 561 questionnaires were collected from 26 pilot survey sites. The awareness rate for A. lumbricoides (62.78%, 981/1 561) was higher than that for Clonorchis sinensis (27.16%, 424/1 561)(P < 0.05). The awareness for parasite infections and healthy behavior in people living in urban cites are better than those who live in rural areas. Conclusion The infection rate of common parasite infections in Tianjin is low. Most people are aware of soil-transmitted nematodes and proper healthy behaviors, less aware of clonorchiasis.

National surveillance of soil-transmitted helminth infections in 2017

Hui-hui ZHU, Ji-lei HUANG, Ting-jun ZHU, Lei DUAN, Chang-hai ZHOU, Men-bao QIAN, Ying-dan CHEN

2019, 37(1):  12-17.  doi:10.12140/j.issn.1000-7423.2019.01.003

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Objective To determine the nationwide epidemic status of soil-transmitted helminth (STH) infections and the prevalent risk factors in China, so as to provide scientific basis for evaluating the existing control efforts and formulating control strategies. Methods The surveillance for STH was carried out in 291 surveillance sites or counties in 30 provinces/municipalities/autonomous regions in China in 2017. Each surveillance site includes 5 villages evenly distributed geographically, and 200 permanent residents with age of over 3 years old were randomly enrolled from each village. The stool samples were collected from each enrolled villager and the eggs of STH were examined under microscope in duplicate using modified Kato-katz thick smear method. The prevalence for STH infections were then calculated. Chi-square test was used to compare the infection rates among different groups. In addition, the contamination of Ascaris eggs and hookworm larvae was also monitored in soil samples collected from 5 households in each village. Results Total 297 078 people were enrolled in the surveillance from the total 291 surveillance sites and the overall prevalence of STH infections was 1.78% (5 293/297 078)nationwide. The top three provinces with highest prevalence of STH infections include Yunnan (11.83%, 1 345/11 372), Hainan (10.91%, 220/2 017)and Chongqing (9.68%, 355/4 091). The prevalence of STH in female villagers (1.94%, 2 945/151 771)was significantly higher than male (1.62%, 2 348/145 307)(P < 0.01). Villagers with age of 60 years old and above had the highest infection rate of 2.78%(1 986/71 314), followed by 7-14 age group(1.71%)and 45-59 age group(1.69%), with statistical difference among the different age groups(P < 0.01). The overall prevalence of hookworm, Ascaris lumbricoides, and Trichuris trichiura within the investigated villagers were 1.00%, 0.50% and 0.41%, respectively. However, most of them were light infection, accounting for 93.85%(2 791/2 794)for hookworm, 82.89%(1 221/1 473)for A. lumbricoides and 90.20%（1 104/1 224）for T. trichiura. The moderate infection account for 3.33%(99/2 794) for hookworm, 16.50%（243/1 473）for A. lumbricoides and 9.6%(117/1 224) for T. trichiura. The heavy infections only accounted for 2.82%(84/2 794)for hookworm, 0.61%(9/1 473)for A. lumbricoides and 0.25%(3/1 224)for T. trichiura. The contamination rates of Ascaris eggs and hookworm larvae in collected soil were 5.59% (114/2 040) and 3.18% (64/2 040), respectively. Conclusion Based on the nationwide surveillance of STH from 291 surveillance sites in 2017 the overall prevalence of STH in China was low, however, the STH prevalence in some provinces with low economic development still remained high and the endemic risks still existed. It is needed to continue the surveillance and strengthen the control efforts especially in those areas with low economic growth.

Leishmania infantum infection stimulates neutrophils to generate extracellular traps in mouse

Fu-rong WEI, Yue-tao YANG, Jun-yun WANG, Yan-juan WANG, Jia-ming PAN, Jian-ping CAO

2019, 37(1):  18-22.  doi:10.12140/j.issn.1000-7423.2019.01.004

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Objective To analyze the effect of Leishmania infantum infection on the differentiation of neutrophil extracellular traps (NETs) of mouse neutrophils under induction of MPA and the possible functions associated with the defense mechanism or pathogenesis of leishmaniasis. Methods Each of 30 BABL/c mice was intraperitoneally infected with 1 × 10⁸ L. infantum promastigotes in total volume of 0.5 ml. The peripheral blood were collected from the ether-anesthetized mice infected with L. infantum for 12 weeks or from normal control mice and the blood cells were stimulated with 100 nmol/L PMA for 3 h to generate NETs. The Red blood cells were lysed and the neutrophils were co-labeled with CD11b and Ly6G and NETs-generating neutrophils were labeled with H3Cit (citrullinated histone 3) and MPO (myeloperoxidase). The neutrophils and NET-generating neutrophils (H3Cit⁺MPO⁺) were quantitatively analyzed by flow cytometry. For fluorescent staining, the neutrophils were isolated from peripheral blood and the NET-generating neutrophils were induced with 100 nmol/L PMA for 3 h. The DNA, H3Cit and MPO in the neutrophils were stained with specific antibodies labeled with blue, red and green fluorescence, respectively and visualized under laser scanning confocal microscope. Results L. infantum infection significantly stimulated the differentiation of H3Cit⁺MPO⁺ NET neutrophils in the blood of mice after being stimulated with PMA (1.880 + 0.251)% compared to that in the normal control mice (0.623 + 0.087)% with statistical significance (P < 0.01). Laser confocal microscopic imaging analysis showed that the L. infantum infection stimulated more neutrophils of mouse to differentiate to NETs characterized by the filamentous DNA and H3Cit⁺MPO⁺ particles attached to the DNA skeleton structure, compared to neutrophils from normal mouse.

Rapid detection of Schistosoma japonicum specific gene fragment by recombinase aided isothermal amplification combined with fluorescent probe

Song ZHAO, Yan-hong LIU, Ting LI, Wei LI, Jian-feng ZHANG, Li-chuan GUO, Qing-jie YING, Hai-tao YANG, Kun YANG

2019, 37(1):  23-27.  doi:10.12140/j.issn.1000-7423.2019.01.005

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Objective To establish a novel and rapid method to detect Schistosoma japonicum specific gene fragment using a real-time recombinase aided isothermal amplification (RAA) combined with a fluorescent probe. Methods S. japonicum G28（SjG28） was selected as the target gene to be detected, and the primers and fluorescent probes were designed accordingly. A real-time fluorescent RAA using RAA combined with fluorescent probe was established to detect SjG28. The sensitivity and specificity of the real-time fluorescent RAA was determined and optimized by using gradient dilutions of SjG28 recombinant plasmid as template and the genomic DNAs from other helminths of S. mansoni, Echinococcus granulosus, Ancylostoma duodenale, Ascaris lumbricoides and Clonorchis sinensis as controls. Then this method was applied to detect SjG28 in genomic DNA extracted from snails and feces of rabbits infected with S. japonicum. Results As low as 10 copies of SjG28 plasmid DNA could be detected by the developed real-time fluorescent RAA within 10 min. This method was able to detect SjG28 in the fecal samples from rabbit infected with different dose of cercarias (20, 200 and 500) within 15 min and snail pool (50) mixed with 1, 2, 3, 4, 5 and 10 cercaria-positive snails within 10 min. There was no product amplified from genomic DNAs extracted from other helminths: S. mansoni, E. granulosus, A. duodenale, A. lumbricoides and C. sinensis. Conclusion A novel and rapid florescent real-time fluorescent RAA method was established to detect S. japonicum with high sensitivity and specificity.

Infection status and molecular identification of Clonorchis sinensis in human population of Tengxian County, Guangxi

Meng XU, Wen HUANG, Shen OU, Jian-hai YIN, Sheng-kui CAO, Li-yu MENG, Jian-ping CAO, Yu-juan SHEN

2019, 37(1):  28-32.  doi:10.12140/j.issn.1000-7423.2019.01.006

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Objective To understand the infection status and identify the isolates of Clonorchis sinensis in human population in Tengxian County, Guangxi Zhuang Autonomous Region, in order to provide base information for the control of clonorchiasis. Methods Two villages from Tianping Town and Jinji Town were selected for the survey by stratified cluster random sampling method. Demographic and relative information were collected through a questionnaire survey. Fecal samples were collected from villagers with age over 3 year-old and eggs of C. sinensis in fecal samples were microscopically examined using a modified Kato-Katz technique in triplicate. The information for the distribution of C. sinensis infection in residents with different gender, age and education was analyzed. Egg-positive fecal samples were selected for DNA extraction. The fragment of internal transcribed spacer 2 (ITS2) of C. sinensis was amplified using PCR for DNA sequencing. The obtained sequences were compared with those deposited in GenBank. Results A total of 400 individuals in the two villages were enrolled for investigation, of which 208 were infected with C. sinensis. The prevalence of C. sinensis was 52.0% (208/400), majority of them were mild infection (69.2%, 144/208). The prevalence in Tianping Town (66.5%, 133/200) was higher than that in Jinji Town(37.5%, 75/200) with statistical significance (χ² = 33.69, P < 0.01). The prevalence in males (58.7%, 131/223) was significantly higher than that in females (43.5%, 77/177) (χ² = 9.18, P < 0.01). The highest prevalence appeared in the 30~39 years group (63.8%, 60/94). Individuals who received senior high school education and above had the highest prevalence (68.0%, 17/25). The prevalence was significantly different among the age groups (χ² = 37.1, P < 0.01), as it was for different levels of education (χ² = 23.02, P < 0.01). The sequence of 156 bp ITS2 fragment amplified from the DNA isolated from the fecal eggs was identical to the isolate of C. sinensis from Vietnam Da Nang (accession number: MF319655). Conclusion The overall prevalence of C. sinensis in Tengxian County was 52.0%, a heavily endemic area for clonorchiasis. The infection was preferable in males with middle age and higher education. The sequence of C. sinensis ITS2 isolated from this County was identical to that of a Vietnam Da Nang isolate.

SHORT COMMUNICATIONS

Endemic status of malaria in Nanyang, Henan, during 2010-2017

Ting-zhu WANG, Wen-tao ZHANG, Hong-pu ZHANG

2019, 37(1):  32-35.  doi:10.12140/j.issn.1000-7423.2019.01.020

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The malaria endemic characteristics in Nanyang, Henan, during 2010-2017 was analyzed in order to formulate some strategies for consolidating malaria elimination in the region. Through the disease surveillance information reporting system of the Chinese Center for Disease Control and Prevention, the epidemiological data of malaria cases in Nanyang, Henan, during 2010-2017 were collected and analyzed for the characteristics of epidemic situation, regional distribution, gender, age and occupational distribution, source of infection, time to get diagnosed and treated and where the case reported from using Excel 2003. The epidemiological results showed that total 375 malaria cases were reported in Nanyang, Henan, from 2010 to 2017, of which 181 cases were clinically diagnosed (48.3%) and 194 cases diagnosed in the laboratory (51.7%). 306 cases were caused by the infection of Plasmodium vivax (81.6%), 61 cases by P. falciparum infection (16.3%), 7 cases by P. Ovale (1.9%), 1 case by P. malariae (0.2%). There were 299 local infections (79.7%) and 76 imported cases (20.3%). P. vivax was the dominant species that caused malaria in Nanyang area during 2010-2011, with local infection and clinically diagnosed cases only. During 2012-2017, there was no local infection anymore and all cases were imported from abroad or other endemic areas and diagnosed in laboratories only. Malaria cases were identified in 13 counties or cities in Nanyang area, with the largest number of cases identified in Tanghe County (61 cases) and the lowest number of cases in Xixia County (5 cases). In the infected patients, males accounted for 69.1% (259/375) and females 30.9% (116/375). The infected cases were identified in people with age from 1 to 86 years old. The number of infected cases in the 40-49 and 50-59 age groups was 24.3% (91/375) and 18.9% (71/375), respectively. The number of infected cases in the 0-9 group was the lowest, accounting for 2.1% (8/375). Malaria cases were mainly seen in local farmers during 2010-2011, then shifted to migrant workers during 2012-2017. In the seasonal distribution, the peak number of local malaria cases in Nanyang during 2010-2011 was observed during May-October, accounting for 80.3% (240/299). However, there was no significant seasonal distribution of 76 imported malaria cases during 2010-2017. Sources of imported malaria cases include Africa (90.7%), Southeast Asia (4.0%), South Asia (4.0%), and Yunnan-Myanmar border (1.3%). The top five countries from which the most imported malaria cases came were Nigeria (21 cases), Angola (11 cases), Cameroon (9 cases), Equatorial Guinea (5 cases) and Uganda (5 cases), accounting for 67.1% of total imported cases (51/76). Since the launch of the Malaria Elimination Action in September 2010, the average time from the onset of malaria to diagnosis has been shortened to 4 days, and the time from diagnosis to the case reporting has been shortened to 1 day. During 2010-2011, most of the malaria cases were diagnosed and reported by Township Clinic (68.4%, 216/316). From 2012 to 2017, the malaria cases were mainly diagnosed and reported by the medical institutions at and above the municipal level (86.4%, 51/59).

ORIGINAL ARTICLES

Endemic status of human Ascaris lumbricoides infection in Jiangxi Province

Zhao-jun LI, Wei-ming LAN, Jun GE, Yue-ming LIU, Fang-yu YI, Xiao-jun ZENG, Wei-sheng JIANG, Chun-qin HANG, Jing XU, Shan LV, Shi-zhu LI

2019, 37(1):  36-40.  doi:10.12140/j.issn.1000-7423.2019.01.007

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Objective To understand and analyze the prevalence and endemic status of Ascaris lumbricoides infection in Jiangxi province, so as to provide scientific basis for the development of control strategy. Methods A survey was performed in Jiangxi Province according to the protocol detailed by the 3rd National Investigation of Human Key Parasitic Infections in 2014. Total 92 survey sites in 32 counties were chosen using stratified cluster sampling method based on the different ecological, economical and geographical status. More than 250 permanent residents with age over 3 years old were involved in each site. The stool samples were collected from each participant and the eggs of A. lumbricoides in the stool samples were microscopically examined by Kato-Katz technique in duplicate. The difference in infection rate was compared between different groups of participants using the chi-square test. Results A total of 23 606 villagers were surveyed and the overall prevalence of A. lumbricoides infection among the investigated population was 0.87% (206/23 606). The infection of A. lumbricoides was found in the 20 of 32 surveyed counties with highest prevalence in Yudu (5.35%, 42/785), which is statistically different compared with prevalence in other surveyed counties (P < 0.01). For the prevalence in different ecological regions, the prevalence in Nanling ecological region(1.65%, 78/4 742) was significantly higher than that in Zhejiang/Fujian ecological region (0.10%, 6/5 887)（P < 0.05). The prevalence of A. lumbricoides infection in male villagers was 0.75% (86/11 538) that is not significantly different compared with the prevalence in female (0.99%, 120/12 068)(P > 0.05). Among 206 infected people, 97 were identified with fertilized eggs (47.09%), 109 were identified with unfertilized eggs (52.91%), and 46 were identified with mixed fertilized and unfertilized eggs. The infection was found in all age groups with the highest infection rate in the 30-39 year group(1.14%, 28/2 465). For the occupation distribution, the infection rate was highest in businessman (1.50%, 4/267) and lowest in unemployed people (0). In terms of education, the infection rate was highest in the people with illiteracy (1.13%, 43/3 818), and lowest in people with high school (0.49%, 8/1 631). There is statistical difference among the people with different age groups, occupations, and education level, respectively (P < 0.01). Conclusion The prevalence of A. lumbricoides has dropped to the lowest level in Jiangxi Province. More unfertilized eggs were identified than fertilized eggs in the infected people. However, the regional distribution of A. lumbricoides infection was regionally discrepant with some areas with high infection, which is still one of the key issues for the control of ascariasis in Jiangxi Province.

Audit and quality assessment on malaria case information reported by National Information Management System for Malaria in China in 2017

Hong TU, Jun FENG, Li ZHANG, Shao-sen ZHANG, Zhi-gui XIA, Shui-sen ZHOU

2019, 37(1):  41-47.  doi:10.12140/j.issn.1000-7423.2019.01.008

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Objective To audit and assess the credibility of malaria case information reported by National Information Management System for Malaria in 2017 and identify the problems and gaps in the malaria case report network, and provide the corresponding suggestions for improving the system accordingly. Methods The malaria cases reported by National Information Management System for Malaria from 24 malaria-endemic provinces and the case information were audited and re-assessed by National Technical Committee for Malaria Elimination. The audit results for the report credibility of malaria cases in different provinces, different seasons and different regions, and their influences were statistically analyzed using descriptive statistics, chi-square tests and trend tests. Results A total of 2 651 malaria cases reported in 24 malaria endemic provinces (municipalities, autonomous region) in 2017 and the case information were audited and re-assessed by the committee experts, only 58.17% (1 542/2 651) of which were qualified through the preliminary assessment. The preliminarily assessed qualified rate for each case of quarter were 58.18% (377/648), 61.73% (450/729), 48.85% (360/737) and 66.11% (355/537), respectively, with no significant linear trend(P > 0.01). The qualified rate of preliminary assessment for cases collected from Eastern, Central and Western endemic regions were 55.96% (856/1 529), 60.35% (312/517) and 61.82% (374/605), respectively, without significant difference among them. The three provinces with the lowest assessment qualification rate were Hubei (36.00%, 36/100), Guizhou (37.50%, 9/24) and Hebei (40.68%, 24/59). The main reasons for the unqualified case report included the lack of provincial-level re-assessment (29.23%), insufficient evidence for case diagnosis (5.17%), incomplete case information (5.28%) and irregular antimalarial medication(2.15%). After being re-assessed through adding and collecting more information, the qualified rate of reported malaria cases increased to 92.49% (2 452/2 651) and the above mentioned unqualified reasons dropped to 3.06% (lack of provincial-level re-assessment), 2.04% (insufficient evidence), 1.36% (incomplete information) and 1.06% (irregular treatment), with statistical significance compared to the preliminary assessment results (P < 0.01). The re-assessment also increased the qualified rate of malaria cases reported in each quarter with 89.81% (582/648) for the 1st quarter, 91.08% (664/729) for the 2nd quarter, 93.08% (686/737) for the 3rd quarter and 96.83% (520/537) for the 4th quarter with linear trend (P < 0.01). However, there was no statistically significant difference in the unqualified rate of re-assessment in different regions (P > 0.01). The re-assessed qualified rate for cases reported in the eastern, central and western regions increased to 93.07% (1 423/1 529), 90.72% (469/517) and 92.56% (560/605) respectively, with no statistically significant difference among the different regions (P > 0.01). However, the rate of irregular antimalarial medication in western region (2.51%, 13/605) was higher than that in the central (1.93%, 10/517) and eastern (0.33%, 5/1 529) regions with statistical significance (P < 0.01), whereas the other unqualified reasons had no statistical difference among different regions (P > 0.01). The three provinces with the lowest qualified rate of malaria case report after being re-assessed were Guizhou (75.00%, 18/24), Henan (82.49%, 76/177) and Gansu (15/18). Conclusion Re-assessment of reported national malaria cases and case information have improved the qualification and accuracy of malaria cases reported by National Information Management System for Malaria. The main reason for the inaccuracy is the irregularities during the case information report.

Epidemiological analysis on malaria cases reported in Shandong Province during 2015-2017

Yan XU, Yong-bin WANG, Chang-lei ZHAO, Xiu-qin BU, Long-jiang WANG, Xiang-li KONG

2019, 37(1):  48-54.  doi:10.12140/j.issn.1000-7423.2019.01.009

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Objective To understand epidemiological characteristics of malaria cases reported in Shandong Province, so as to provide scientific evidence for better prevention and control of malaria. Methods The data of malaria reported in Shandong Province during 2015-2017 were collected from the Information Management System for Infectious Diseases Report and Information Management System for Parasitic Diseases Control and Prevention, and the clinical manifestations of malaria cases were collected and statistically analyzed by SPSS 18.0. Results Among 676 malaria cases reported during 2015-2017, 99.85% (675/676) were imported and most of them were imported from African countries (96.75%, 654/676), mainly from Equatorial Guinea（143, 21.87%）and Angola（85, 13.00%）. All cases were laboratory confirmed with 76.04% of Plasmodium falciparum infection and 9.76% of P. vivax infection. About 1/4 patients (28.85%, 195/676) went to visit doctor on the first day with symptoms, while 38.17% (258/676) of patients went to visit doctor three or more days after they had symptoms. About 1/3 of patients were diagnosed as malaria at the first doctor visit (37.43%, 253/676) and 1/3 misdiagnosed as other diseases(31.66%, 214/676), and another 1/3 were diagnosed as malaria 3 days after the first visit (32.84%, 222/676). The accuracy for diagnoses of P. falciparum, P. vivax, P. ovale and P. malariae infections were 97.49%(504/517), 67.86% (57/84), 90.91% (50/55) and 91.67% (11/12), respectively, after being re-confirmed by provincial experts. Among 65 severe malaria patients only 17 went to see doctor on the first day with clinical symptoms(26.15%) and 18 patients were diagnosed as malaria at the first visit (27.69%). 17 cities reported malaria cases during the period (2015-2017) and 74.26% (502/676) of the total malaria cases were reported in 6 cities (Jining, Tai’an, Weihai, Yantai, Qingdao and Jinan). The imported cases were reported in all 17 cities and most of them (59.32%, 401/676) distributed in 5 cities (Tai’an, Yantai, Weihai, Jining and Qingdao). More young and middle-aged men were infected. The minimum age was 16 years old, the maximum age was 65, and the median age was 41 years old. They were mainly peasants（422,62.43%）. Conclusions Total 676 malaria cases were reported in Shandong Province during 2015-2017 and 96.75% of them were imported from Africa. The rates for going to see doctor on the first day with clinical symptoms and the accurate diagnosis with malaria on the first visit were low.

Epidemiological analysis of malaria cases reported in Shanghai from 2010 to 2017

Min ZHU, Li CAI, Chen-gang ZHANG, Yao-guang ZHANG, Zhen-yu WANG, Li JIANG, Xiao-jiang MA, Qian ZHU, Yan-yan HE

2019, 37(1):  55-60.  doi:10.12140/j.issn.1000-7423.2019.01.010

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Objective To understand the epidemiological characteristics of malaria cases reported in Shanghai from 2010 to 2017, so as to provide evidence-based reference for adjusting strategies and measures in the process of malaria elimination in Shanghai. Methods The epidemiological data of malaria cases reported in Shanghai from 2010 to 2017 were collected. Classification, diagnosis time, infection source and epidemiological characteristics of the cases, were statistically analyzed by using Microsoft Excel 2010. Results A total of 359 malaria cases were reported in Shanghai from 2010 to 2017 with 3 deaths. Among them 10 patients were clinically diagnosed (2.8%, 10/359) and the rest were laboratory diagnosed (97.2%, 349/359). 241 were infected with P. falciparum (67.1%, 241/359), 60 with P. vivax (16.7%, 60/359), 26 with P. ovale (7.2%, 26/359), 13 with P. malariae (3.6%, 13/359) and 9 infected with unknown species (2.5%, 9/359). All cases were imported and there was no indigenous case reported in Shanghai since 2010. The median diagnosis time was 4.4 days after the clinical symptoms occurred, 47.8% (137/359) cases were diagnosed within 3 days. There was no significantly seasonal difference in the case report. Young and middle-aged men were preferably infected. The case distribution included 105 Shanghai native residents (29.2%, 105/359), 207 from other provinces (57.7%, 207/359) and 47 foreigners (13.1%, 47/359). The current living address of cases distributed in 92 counties in 19 provinces of China, and 261 cases (72.70%, 261/359) were Shanghai residents distributed in 16 districts of Shanghai. 338 cases (94.2%, 338/359) were imported from overseas, and 19 cases (5.3%, 19/359) were imported from other provinces of China. Since 2012 all reported cases were imported from overseas except for one case of P. malariae transmitted by blood transfusion and one case of P. malariae without definite source of infection. Conclusion No indigenous malaria case has been reported in Shanghai since 2010. However, there are still imported malaria cases mainly from overseas so that we need to pay attention to malaria surveillance in Shanghai.

Application of high-throughput DNA microarray for rapid detection of Plasmodium spp. at port

Ji-hong CAO, Zi-long ZHANG, Shen-wei LI, Mei LI, Xiao-hang ZHANG, Zhen-gan TIAN

2019, 37(1):  61-65.  doi:10.12140/j.issn.1000-7423.2019.01.011

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Objective To validate the application of a sensitive and specific high-throughput DNA microarray chip method for rapid detection of four species of Plasmodium simultaneously. Methods The blood samples with infections of different species of Plasmodium (P. vivax, P. falciparum, P. ovale, P. malariae) identified by microscopy and fluorescent PCR were collected from quarantined passengers at airport, and used for validating the sensitivity and specificity of the high-throughput DNA microarray chip to detect Plasmodium spp.. DNAs were extracted from the blood samples and quantified by the quantitative real-time PCR (RT-PCR). The sensitivity of the DNA microarray was determined by the series dilutions of extracted DNA samples. The specificity of the DNA microarray was determined by cross reacting with the specific Plasmodium sample mixed with the same amount of DNA extracted from P. knowlesi, Cryptosporidium parvum and Balantidium coli. The microarray chip was also used for detecting 50 blood samples collected from patients with suspected malaria from the border area of Yunnan Province, compared with results of microscopy. Results The outcome of the DNA microarray detection of plasmodium spp. completely matched the results detected by microscopy and PCR. The whole procedure only took 2 hours. The chip was able to detect as low as 78 copies/μl of P. vivax, 87 copies/μl of P. malariae, 135 copies/μl of P. falciparum and 302 copies/μl of P. ovale. There was no cross-reaction with P. knowlesi, C. parvum and B. coli. The microarray chip was used to detect 50 blood samples from suspected malaria patients from Yunnan border, 18 of them was identified as infection with P. vivax, 12 for P. falciparum, and 1 for P. malariae. The chip results were totally consistent with the microscopic results. Conclusion The high-throughput DNA microarray is a rapid, sensitive and specific method to detect different species of Plasmodium infections with no cross-reaction to other species of protozoans.

Analysis of molecular epidemiology of babesiosis in patients having fever and thrombocytopenia in Xinyang City, Henan Province

Su-hua LI, Yu-ling ZHAO, Li-jun GAO, Ya-lan ZHANG, Rui-min ZHOU, Dan QIAN, Cheng-yun YANG, Ying LIU, De-ling LU, Hong-wei ZHANG, Bian-li XU

2019, 37(1):  66-69.  doi:10.12140/j.issn.1000-7423.2019.01.012

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Objective To study the molecular epidemiology of babesiosis in cases of fever with thrombocytopenia in Xinyang City and provide basis for the control of babesiosis. Methods Blood samples and epidemiological information were collected from 600 patients who had fever and thrombocytopenia in Xinyang City in 2012. The DNA was extracted from the blood samples and Babesia 18S rRNA was amplified by nested-PCR with two sets of primers Bab5/Bab8 + Bab6/Bab7 and RIB-19/RIB-20 + BAB-rumF/BAB-rumR. The PCR products were sequenced and aligned with Babesia mocroti sequences in genome database. The PCR positive cases were further analyzed for their epidemiological characteristics, history of tick bite and bunyavirus infection. Results Totally 600 blood samples were collected from patients who had fever and thrombocytopenia, 59 (9.8%) of them were PCR positive with a fragment of 430 bp amplified by both sets of primers. The amplified 18S rRNA sequences shared more than 97% identity to LC314655.1 of B. mocroti. Among the 59 positive cases, 51 (86.4%) were from Xinyang City, 16 from Shangcheng County (27.1%) and 12 from Guangshan County (20.3%). The fever onset mostly happened in summer including 14 in May (23.7%), 23 in June (39.0%) and 10 in July (17.0%). Most of cases were middle-aged or elderly and 48 cases (81.4%) were over 40 years old. Among the 59 positive cases 14 (23.7%) had history to be bitten by ticks and 31 (52.5%) were positive for bunyavirus. Conclusion The morbidity of babesiosis is high in the cases with fever and thrombocytopenia in Xinyang City, Henan Province. People with middle-age and elderly are the most vulnerable population to be infected by Babesia during summer.

Cloning and expression of 45 kDa Taenia asiatica estrogen-regulated protein and development of monoclonal antibodies

Li-qun WANG, Pan-hong LIANG, Shao-hua ZHANG, Jun-ling HOU, Li-jie WANG, Li MAO, Xue-nong LUO

2019, 37(1):  70-74.  doi:10.12140/j.issn.1000-7423.2019.01.013

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Objective To clone and express 45 kDa Taenia asiatica estrogen-regulated protein (TaEP45) as a diagnostic antigen for detecting infection of T. asiatica. Methods The open reading frame of TaEP45 was amplified from the total cDNA of T. asiatica using specific primers designed from TaEP45 sequence deposited in GenBank, then subcloned into the prokaryotic expression vector pET30a(+). The recombinant plasmid DNA was transformed into Escherichia coli BL21（DE3） cells for expression. The recombinant TaEP45 with His-tag was expressed in E. coli under induction of 1 mmol/L isopropyl β-D-thiogalactoside(IPTG) and purified with nickel column. The purified recombinant TaEP45 was tested for its recognition by serum from pig infected with T. asiatica by Western blotting, and then used to immunize BALB/c mice to make monoclonal antibody (McAb). Results RT-PCR amplified a gene product of 1 362 bp which encodes 453 amino acids of TaEP45. The recombinant TaEP45 was highly expressed in E. coli BL21 cells with relative molecular mass of 60 000. The purified recombinant TaEP45 was strongly recognized by serum from pig infected with T. asiatica. After being immunized with recombinant TaEP45 formulated with Freud’s adjuvant, the splenocytes of mice were used to make hybridoma cell lines. Total 5 hybridoma cell lines were obtained that stably produced McAbs against TaEP45 with titers higher than 1 : 2 430. The subtype antibodies were IgG2b or IgG1. All McAbs were able to recognize T. asiatica worm lysates and recombinant TaEP45 as well. Conclusion The TaEP45 was successfully cloned from T. asiatica cDNA and the recombinant TaEP45 protein was expressed in E. coli. Five hybridoma cell lines that stably secreted McAbs against TaEP45 are obtained with high titer of antibodies, which are able to recognize T. asiatica worm lysates.

Species identification of Pomacea snails in southwest Yunan Province based on COⅠgene polymorphism

Cang-lin ZHANG, Xue-ying BAO, Jia PENG, Jin-rong ZI, Zhen RAN, Na LU, Ya-ming YANG

2019, 37(1):  75-81.  doi:10.12140/j.issn.1000-7423.2019.01.014

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Objective To identify Pomacea spp. based on the polymorphism of mitochondria DNA cytochrome c oxidase subunit I gene (COⅠ) amplified from Pomacea collected from southwest Yunnan Province. Methods The samples of Pomacea snails were collected from Jinghong City and Mengla County of Xishuangbanna, Simao District and Ximeng County of Pu’er, Gengma County of Lincang, Ruili City of Dehong, Yunnan Province, from November 18, 2016 to May 8, 2017, morphological observation was performed. DNA was extracted and COⅠgene was amplified and sequenced. The phylogenetic tree and genetic distances among the snail samples compared with known species of Pomacea were constructed using the neighbour-joining method with MEGA software (version 6.06). Results A total of 200 Pomacea snails were collected. It is difficult to distinguish species by morphological identification. 129 COⅠsequences were amplified, 123 sequences belonged to Pomacea canaliculata, 6 sequences belonged to P. insularum. The final length of amplified sequence was 621 bp, there were 76 nucleotide variation sites and 76 simplified information sites, without single variation, insertion and deletion sites, and without base conversion and transmutation. The average contents of A, G, T and C were 23.1%, 20.5%, 39.8% and 16.7%, respectively. The DNA sequence for 207 amino acids with 203 conserved (98.1%). The results showed that COⅠ gene sequence fell into 4 unique haplotypes based on the single mutations. HAP-1 haplotype was the predominant one (71.3%, 92/129), followed by HAP-3(17.8%, 23/129), HAP-2 (6.2%, 8/129） and HAP-4 (4.7%, 6/129). The sequence data showed that HAP-1, HAP-2, HAP-3 shared closer genetic distance to P. canaliculata ranged from 0.000 to 0.052 (with 0 or 31 nucleotide differences), further genetic distance to Pila conica with distances ranged from 0.214 to 0.239 (with 115-126 nucleotide differences). The genetic distances between HAP-4 and P. canaliculate, P. insularum, Pila conica were 0.100, 0.060 and 0.223, and with nucleotide differences of 57, 35 and 119, respectively. The genetic distances among the four HAPs ranged from 0.021-0.100 with nucleotide differences of 13-57. Among them, HAP-1 is closer to HAP-2 (0.021), HAP-4 is further away from HAP-1, HAP-2 and HAP-3 (0.100). The genetic distance between HAP-3 and HAP-1/HAP-2 was 0.052. HAP-1, HAP-2 and HAP-3 shared 98%-100% sequence identity to Pomacea canaliculate (GenBank accession AB433769, KT313034、EU523129), and HAP-4 shared 99% sequence identity to P. insularum (EF514942). Conclusion The species of Pomacea snails in southwest Yunnan Province are P. canaliculata and P. insularum based on COⅠgene polymorphism with certain genetic diversity amongst the acquired haplotypes of P. canaliculata identified in this study.

Sequence polymorphisms within Kalicephalus indicus ribosomal and mitochondrial genes

Cheng-yan ZHOU, Jun MA, Wei LIU, Qian-ming XU, Xing-quan ZHU

2019, 37(1):  81-86.  doi:10.12140/j.issn.1000-7423.2019.01.015

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Objective To analyze the ribosomal and mitochondrial genes sequences polymorphisms of Kalicephalus indicus, and discuss their application potential as molecular markers. Methods Two wild Elaphe carinata snakes were captured in June 2016, in Zhangli Town, Changsha County, Hunan Province, and seven K. indicus adult worms were isolated from the snakes. Total DNA from the worms was extracted. The sequences of the first and second internal transcribed spacer regions 1 and 2(ITS-1 and ITS-2), 5.8S, mitochondrial cytochrome c oxidase subunit 1 (cox1), small subunit 12S of ribosomal RNA, ATP synthase subunit 6 (atp6), nicotinamide adenine dinucleotide dehydrogenase subunit 4 (nad4) of K. indicus were amplified using PCR and subjected to DNA sequencing. The sequencing results of each gene were aligned using ClustalX 1.83. A phylogenetic tree based on the mitochondrial DNA sequences cox1, 12S, atp6 and nad4 was constructed using Bayesian method, using Onchocerca volvulus as an outgroup. Intraspecific variation in these sequences among the seven K. indicus samples were analyzed. Results The size of the amplified products of ITS-1, 5.8S and ITS-2 was approximately 900 bp. The size of the amplified products of cox1, 12S, atp6 and nad4 was approximately 400, 600, 500 and 400 bp, respectively. The content of A + T of ITS-1, 5.8S and ITS-2 is 52.7%-53.7%, higher than its content of G + C (46.3%-47.4%). The content of A + T of cox1, 12S, atp6 and nad4 is 67.1%-68.9%, 76.9%-77.4%, 73.0%-75.5%, and 70.3%-73.3%, respectively, higher than their content of G + C (31.1%-32.9%, 22.6%-23.1%, 24.5%-27.0% and 26.7%-29.7%). Intraspecific variation in ITS-1, 5.8S, ITS-2, cox1, 12S, atp6 and nad4 were 0-2.5%, 0, 0-5.0%, 0-2.6%, 0-3.8%, 0.7%-8.7% and 0-11.0%, respectively. The phylogenetic tree clustered the seven K. indicus individuals into the same branch, with Ancylostoma ceylanicum being the closest species. Conclusion The ITS-2 sequence with the largest variation is expected to be an ideal molecular marker for studying the intraspecific variation of K. indicus. The degree of its intraspecific variation in mitochondrial gene sequences is as follows: cox1 < 12S < atp6 < nad4, and nad4 is an ideal mitochondrial molecular marker for studying the genetic diversity of K. indicus.

Prediction of Cryptosporidium spp. infection in rural residents of Binyang County, Guangxi by non-equidistant grey model GM(1,1)

Ning XU, Lin-hua TANG, Yan-yan JIANG, Yu-juan SHEN, Sheng-kui CAO, Hua LIU, Jian-hai YIN, Yi-chao YANG, Zhi-hua JIANG, Wei LI, Xiao-qin GAN, Jia-guang ZHAO, Wei-jie ZHENG, Li WANG, Rong ZHANG, Jian-ping CAO

2019, 37(1):  87-91.  doi:10.12140/j.issn.1000-7423.2019.01.016

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Objective To establish a prediction model for Cryptosporidium spp. infection in rural residents of Binyang County, Guangxi Zhuang Autonomous Region (Guangxi) so as to provide reference for the development of prevention and control measures. Methods In 2014, 2016, 2017 and 2018, the rural residents of Binyang County were selected by stratified cluster random sampling, and the fecal samples were collected from selected participants for detecting Cryptosporidium spp. infection using an immunological test strip method. The non-equidistant grey model GM(1,1) was established to predict the prevalence of Cryptosporidium spp. infection in 2019 and 2020 in this region based on the prevalence data collected from the survey in 2014, 2016, 2017 and 2018. The model effect was tested using residual test, correlation test and the posterior deviation test. Results The prevalence of Cryptosporidium spp. infection in the selected rural residents of Binyang County was 2.9% (57/2 000) in 2014, 1.5% (15/1 030) in 2016, 1.1% (11/1 027) in 2017 and 0.6% (6/1 004) in 2018. The prediction model for prevalence of Cryptosporidium spp. infection in this region was established accordingly as: ^（0）（k_{i + 1}） = -（1 - e^{0.289 1Δki + 1}）e^{-0.289 1Δki+ 1}. Based on this model the fitted prevalence of Cryptosporidium spp. infection in this region in 2014, 2016, 2017 and 2018 were 2.9%, 1.5%, 1.0% and 0.7%, respectively, which are closed correlated with the actual prevalence values (r = 0.667 7) with absolute errors of 0.000 0, -0.001 6, 0.136 8 and -0.121 4 and the relative errors of 0.000 0, 0.001 1, 0.124 4 and 0.202 3, respectively, with the posterior variance ratio C = 0.106 8 and the posterior probability P = 1.00. Based on this model, the prevalence of Cryptosporidium spp. infection in Binyang County are predicted as 0.5% in 2019 and 0.4% in 2020. Conclusion A non-equidistant grey model GM(1,1) is established to predict the prevalence of Cryptosporidium spp. infection. The model has high prediction accuracy and good fitting effect. The predicted results show that the infection rate of Cryptosporidium spp. in rural residents of Binyang County is decreasing.

REVIEWS

Research progress on snail gene expression associated with parasite infections

Zhi-yuan YUE, Yun-hai GUO, Zhi-qiang QIN, Yi ZHANG

2019, 37(1):  92-96.  doi:10.12140/j.issn.1000-7423.2019.01.017

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Snails belong to mollusk invertebrate animals widely distributed on land, in fresh and sea water. Some types of snail are the important intermediate hosts of some helminths and the vectors transmiting parasite infections to human. It has been noticed that snails express some specific genes that are associated with the infection of parasite larvae. The differential expression of genes before and after infection of some parasites plays an important role in the interaction between snails and parasites, as well as the sail’s immune defense against parasite infection. This paper reviews the research progress on the differential expression of genes in snails associated with parasite infection, in order to provide references for the related research on the control of snail and snail-transmitted parasitic diseases.

Progress toward development of a vaccine against Echinococcus granulosus infection

Yu-huan WEI, Yuan HU, Jian-ping CAO

2019, 37(1):  97-101.  doi:10.12140/j.issn.1000-7423.2019.01.018

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Cystic echinococcosis caused by infection with Echinococcus granulosus is a serious parasitic zoonosis, which is widely spread in the world. It keeps a big threat to people health and economic development in the epidemic area. Dogs are definitive host of E. granulosus, and cattle, sheep or humans are its intermediate hosts. Because of the limited efficacy of drugs and surgical treatment, development of a vaccine against E. granulosus infection is a very important approach as a supplementary measure to prevent and control the endemic and spread of echinococcosis. This review focuses on the recent progress in the vaccine development against E. granulosus to provide a reference for the further development of a high efficicacy vaccine.

Progress in research on the correlation between Toxoplasma gondii infection and schizophrenia

Jing OUYANG, Yao-kai CHEN

2019, 37(1):  102-106.  doi:10.12140/j.issn.1000-7423.2019.01.019

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Toxoplasma gondii infection is one of the most common opportunistic infections in acquired immune deficiency syndrome patients. In addition to the pathogenesis directly caused by the infection, it has been found that T. gondii infection is related to the onset and progression of schizophrenia in recent years. Many studies have shown that Toxoplasma infection may affect the pathogenesis and progression of schizophrenia by damaging the nervous system, affecting neurotransmitters and the immune system. However, the specific mechanism underlying this correlation is still unclear. Understanding the intrinsic relationship between Toxoplasma infection and schizophrenia will help to interpret the pathogenesis and influencing factors of schizophrenia and provide a new direction for the prevention and treatment of schizophrenia. This article aims to review the progress in research on the correlation between T. gondii infection and schizophrenia.

SHORT COMMUNICATIONS

Epidemiological investigation of a child visceral leishmaniasis case in Wenzhou, Zhejiang Province

Qing-xiang NI, Wei RUAN, Ling-ling ZHANG, Hua-liang CHEN, Li-nong YAO

2019, 37(1):  108-110.  doi:10.12140/j.issn.1000-7423.2019.01.021

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A child case of visceral leishmaniasis was diagnosed in Wenzhou, Zhejiang Province and the epidemiological investigation was performed on this case to identify the cause of infection, clinical manifestation, treatment and prognosis in order to provide a reference for better prevention and control of kala-azar. A patient with 8 years old originally from Qingtian County of Lishui Municipality was hospitalized with symptoms of fever and vomiting started on January 17, 2018. The physical examination found the swollen tonsil, but did not find enlargement of lymph node around the body. Lips were pale. The spleen could be touched 2 cm below the rib. Blood test showed decreased number of white blood cells, red blood cells and platelet, increased ferritin. The activity of natural killer(NK) cells was decreased. Bone marrow smear demonstrated phagocytosis of red blood cells by macrophages. Serological test showed epstein-barr (EB) virus antibody positive, so diagnosed as “EB virus infection-related hemophilic syndrome”. After being treated with VP16, CSA and ganciclovir, the patient got improved in symptoms and discharged from hospital on February 5. The patient was hospitalized on February 19 for the second chemotherapy with VP16 and CSA. The amastigotes of Leishmania donovani was identified in bone marrow hematopoietic smear on March 21. A 120 bp fragment of Leishmania kinetoplast DNA was amplified by PCR. The diagnosis of kala-azar was confirmed. After being treated with standard anti-Leishmania antimony gluconate (intravenous injection of pentavalent antimony 600 mg/day for 6 days, paused for 2 days, followed by pentavalent antimony 300 mg/day for additional 12 days), the symptoms of the patient were improved. It has been confirmed that it was a case of imported kala-azar, most likely infected by the bite of infected sandflies in the park near his home in Spain.

Isolation of single-sex cercaria of Schistosoma japonicum

Meng-tian WEI, Yang WANG, Bi-hui ZHANG, Jun SUN

2019, 37(1):  111-114.  doi:10.12140/j.issn.1000-7423.2019.01.022

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To obtain the single-sex cercaria for relevant research of schistosomiasis a single miracidium or multiple miracidia were used to infect one snail (one-to-one or multiple-to-one models). The released cercaria from each infected snail were used to infect a mouse. Immature adult worms or schistosomula were collected from each infected mouse 3 weeks after infection and the sex of the worms was identified under microscope. The genotype variation was determined by PCR with 6 short tandem repeats (STRs) primers (RRPS, M5A, MPA, 2AAA, MF1 and J5N). Among 4 mice infected with "one-to-one"cercaria, single-sex worms were obtained in all mice, one mouse with female and three mice with male worms. However, among 43 mice infected with cercaria from "multiple-to-one"snails, 19 contained female worms (44.2%), 7 contained male worms (16.3%) and 17 contained mixed-sex worms (39.5%). STR analysis with 6 pairs of microsatellite primers, better than electrophoresis analysis of PCR products, showed the nearly identical genotype for worms collected from "one-to-one" groups and different genotypes for worms collected from "multiple-to-one" groups. Minor genetic variation was detected between single-sex worms collected from "one-to-one" groups possibly due to the mutation(s) occurred during self-reproduction and development in snail. Worms from "multiple-to-one" groups showed significant genetic polymorphism in terms of different STR patterns, even for the single-sex worms compared to those collected from "one-to-one" groups. In conclusion, single-sex cercaria can be obtained by infecting a snail with single miracidium ("one-to-one" model) with nearly identical genotype. Even though the single-sex cercaria are possibly obtained by the multiple-to-one infection model, the genotype is mixed and complicated.