老挝占巴塞省按蚊种类及其疟原虫子孢子感染情况调查

中国寄生虫学与寄生虫病杂志 ›› 2018, Vol. 36 ›› Issue (5): 478-482.

老挝占巴塞省按蚊种类及其疟原虫子孢子感染情况调查

王微莉¹, 杨锐², 罗春海², 郭晓芳², 姜进勇², 周红宁², SomphadSORCHAMPA³, PhoxayVANNAPASEUTH⁴, 张苍林^2,^*()

1 昆明医科大学公共卫生学院,昆明 650500
2 云南省虫媒传染病防控研究重点实验室,云南省疟疾研究中心,金宁一院士工作站,云南省寄生虫病防治所,普洱 665099
3 老挝琅南塔省勐新县医院,老挝勐新县 01000
4 占巴塞省疟疾和寄生虫病中心,老挝巴色县 01000

收稿日期:2018-04-23 出版日期:2018-10-30 发布日期:2018-11-13
通讯作者: 张苍林
基金资助:
中国-大湄公河次区域部分边境地区疟疾/登革热跨境联防联控试点合作项目,澜湄专项基金项目（No. 2020399）

Investigation on species and sporozoite rate of Anopheles spp. in Champasak Province of Laos

Wei-li WANG¹, Rui YANG², Chun-hai LUO², Xiao-fang GUO², Jin-yong JIANG², Hong-ning ZHOU², SORCHAMPA Somphad³, VANNAPASEUTH Phoxay⁴, Cang-lin ZHANG^2,^*()

1 School of Public Health, Kunming Medical University, Kunming 650500, China
2 Yunnan Provincial Key Laboratory of Vector-borne Diseases Control and Research, Yunnan Provincial Center of Malaria Research, JIN Ning-yi Academic Station, Yunnan Institute of Parasitic Diseases, Pu’er 665099, China;
3 Sing District Hospital of Louang Namtha Province, Mengxin County 01000, Laos
4 Malaria and Parasitology Station of Champasak Province, Pakse County 01000, Laos

Received:2018-04-23 Online:2018-10-30 Published:2018-11-13
Contact: Cang-lin ZHANG
Supported by:
Supported by the Pilot Project of Cooperation on Joint Prevention and Control of Malaria/Dengue in Great Mekong Sub-region and China, Lancang?Mekong River Cooperation Fund (No. 2020399)

摘要/Abstract

摘要：

目的对老挝占巴塞省按蚊种类及其疟原虫子孢子感染情况进行调查,为当地疟疾防治措施的制定与评估提供参考。方法 2017年7月在老挝占巴塞省巴通坡县采用诱蚊灯通宵诱蚊法和通宵人诱法进行捕蚊,对捕获的成蚊进行形态学鉴定,取部分雌性按蚊提取基因组DNA,巢式PCR检测恶性疟原虫和间日疟原虫的18S rRNA基因,计算按蚊疟原虫子孢子阳性率。结果共捕获蚊虫34 687只,分属库蚊、按蚊和伊蚊等8个属的29个种。库蚊属为当地优势属,占捕获蚊虫总数的92.6%（32 110/34 687）;其次是按蚊属,占5.6%（1 947/34 687）。迷糊按蚊是按蚊属的优势种,占该属的65.5%（1 275/1 947）;其后是可赫按蚊,占12.1%（235/1 947）;菲律宾按蚊,占11.9%（232/1 947）;中华按蚊,占5.1%（100/1 947）。巢式PCR共检测按蚊336只（可赫按蚊234只、中华按蚊100只和大劣按蚊2只）,8只按蚊间日疟原虫子孢子阳性,阳性率为2.4%（8/336）。其中,可赫按蚊子孢子阳性6只,阳性率为2.6%（6/234）;中华按蚊子孢子阳性2只,阳性率为2.0%（2/100）;均未检测到恶性疟原虫子孢子。巢式PCR扩增片段长120 bp,其序列与间日疟原虫序列（GenBank登录号为：KT991325、KT991317、MG708221、MF540772、LT635613、KU569498、AF145335、KT991314和KX007932）的同源性为99%~100%。结论老挝占巴塞省捕获的可赫按蚊和中华按蚊存在间日疟原虫子孢子自然感染,且阳性率均较高。

关键词: 疟疾, 按蚊, 子孢子, 巢式PCR, 老挝, 占巴塞省

Abstract:

Objective To investigate the species of Anopheles and rate of Plasmodium sporozoites in anopheline mosquitoes in Champasak Province of Laos, so as to provide essential data for malaria control. Methods Mosquitoes were captured by overnight trapping with light traps and by outdoor human landing catches in Batong County of Champasak Province in July 2017. After morphological identification, genomic DNA was extracted from some female adult mosquitoes, and 18S rRNA-based nested-PCR was performed to detect soprozoites of Plasmodium falciparum and P. vivax. The sporozoite positive rate of Anepheles was calculated. Results A total of 34 687 mosquitoes were captured, belonging to 29 species of 8 genera, dominated by the genus Culex (92.6%, 32 110/34 687), followed by genus Anopheles (5.6%, 1 947/34 687). The genus Anopheles was dominated by An. vagus (65.5%, 1 275/1 947), followed by An. kochi (12.1%, 235/1 947), An. philippinensis (11.9%, 232/1 947), An. sinensis (5.1%, 100/1 947). A total of 336 anopheline female mosquitoes (comprising 234 An. kochi, 100 An. sinensis and 2 An. dirus) were examined by nested-PCR, of which 8 were detected to be positive for P. vivax, including 6 An. kochi (positive rate 2.6%, 6/234) and 2 An. sinensis (2.0%, 2/100). No soprozoites of P. falciparum were found. Nested-PCR amplification showed a band of ~120 bp, sequence of which had 99%-100% homology to those of P. vivax (GenBank Accession No: KT991325, KT991317, MG708221, MF540772, LT635613, KU569498, AF145335, KT991314 and KX007932). Conclusion Both An. kochi and An. sinensis mosquitoes carry P. vivax sporozoites with a positive rate of 2.6% and 2.0% respectively.

Key words: Malaria, Anopheles, Sporozoite, Nested-PCR, Laos, Champasak Province

中图分类号:

R384.11

王微莉, 杨锐, 罗春海, 郭晓芳, 姜进勇, 周红宁, SomphadSORCHAMPA, PhoxayVANNAPASEUTH, 张苍林. 老挝占巴塞省按蚊种类及其疟原虫子孢子感染情况调查[J]. 中国寄生虫学与寄生虫病杂志, 2018, 36(5): 478-482.

Wei-li WANG, Rui YANG, Chun-hai LUO, Xiao-fang GUO, Jin-yong JIANG, Hong-ning ZHOU, SORCHAMPA Somphad, VANNAPASEUTH Phoxay, Cang-lin ZHANG. Investigation on species and sporozoite rate of Anopheles spp. in Champasak Province of Laos[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2018, 36(5): 478-482.

图/表 1

参考文献 37

[1]	WHO: World Malaria Report 2017[R]. Report 2017[R]. Geneva: World Health Organization, 2017.
[2]	Kounnavong S, Gopinath D, Hongvanthong B, et al. Malaria elimination in Lao PDR: the challenges associated with population mobility[J]. Infect Dis Poverty, 2017, 6: 81.
[3]	WHO: World Malaria Report 2016[R]. Report 2016[R]. Geneva: World Health Organization, 2016.
[4]	朱韩武, 曹俊, 周华云, 等. 环介导等温扩增技术检测蚊体内间日疟原虫子孢子的研究[J]. 中国血吸虫病防治杂志, 2010, 22(2): 158-163.
[5]	Souris M, Marcombe S, Laforet J, et al. Modeling spatial variation in risk of presence and insecticide resistance for malaria vectors in Laos[J]. PLoS One, 2017, 12: e0177274.
[6]	Jorgensen P, Nambanya S, Gopinath D, et al. High heterogeneity in Plasmodium falciparum risk illustrates the need for detailed mapping to guide resource allocation: a new malaria risk map of the Lao People’s Democratic Republic[J]. Malar J, 2010, 9: 59.
[7]	Mayxay M, Khanthavong M, Chanthongthip O, et al. Efficacy of artemether-lumefantrine, the nationally-recommended artemisinin combination for the treatment of uncomplicated falciparum malaria, in Southern Laos[J]. Malar J, 2012, 11: 184.
[8]	陆宝麟. 中国动物志, 昆虫纲, 双翅目, 蚊科(上卷)[M]. 北京: 科学出版社, 1997.
[9]	陆宝麟. 中国动物志, 昆虫纲, 双翅目, 蚊科(下卷)[M]. 北京: 科学出版社, 1997.
[10]	董学书, 周红宁, 龚正达. 云南蚊类志(上卷和下卷) [M]. 昆明: 云南科技出版社, 2010.
[11]	张苍林, 杨亚明, 叶润, 等. 中缅边境恶性疟原虫裂殖子表面蛋白1, 2基因多态性研究[J]. 国际医学寄生虫病杂志, 2015, 42(3): 121-127.
[12]	Perandin F, Manca N, Calderaro A, et al. Development of a real-time PCR assay for detection of Plasmodium falciparum, Plasmodium vivax, and Plasmodium ovale for routine clinical diagnosis[J]. J Clin Microbiol, 2004, 42: 1214-1219.
[13]	Snounou G, Viriyakosol S, Zhu XP, et al. High sensitivity of detection of human malaria parasites by the use of nested polymerase chain reaction[J]. Mol Biochem Parasitol, 1993, 61: 315-320.
[14]	Alam MS, Chakma S, Khan WA, et al. Diversity of anopheline species and their Plasmodium infection status in rural Bandarban, Bangladesh[J]. Parasit Vectors, 2012, 5: 150.
[15]	周红宁, 张再兴, 李春富, 等. 云南湄公河流域上游河谷地区疟疾媒介传疟作用研究[J]. 中国寄生虫病防治杂志, 2005, 18(6): 407-411.
[16]	刘耀宝, 周华云, 汪圣强, 等. 荧光定量PCR用于按蚊体内疟原虫子孢子检测的研究[J]. 中国血吸虫病防治杂志, 2012, 24(4): 445-449.
[17]	徐大为, 王峰. 蚊媒唾腺疟原虫子孢子的ELISA检测方法[J]. 承德医学院学报, 1998(2): 64-65.
[18]	Bass C, Nikou D, Blagborough AM, et al. PCR-based detection of Plasmodium in Anopheles mosquitoes: a comparison of a new high-throughput assay with existing methods[J]. Malar J, 2008, 7: 177.
[19]	Beier JC, Asiago CM, Onyango FK, et al. ELISA absorbance cut-off method affects malaria sporozoite rate determination in wild Afrotropical Anopheles[J]. Med Vet Entomol, 1988, 2:259-264.
[20]	Lardeux F, Tejerina R, Aliaga C, et al. Optimization of a semi-nested multiplex PCR to identify Plasmodium parasites in wild-caught Anopheles in Bolivia, and its application to field epidemiological studies[J]. Trans R Soc Trop Med Hyg, 2008, 102: 485-492.
[21]	Arez AP, Lopes D, Pinto J, et al. Plasmodium sp.: optimal protocols for PCR detection of low parasite numbers from mosquito (Anopheles sp.) samples[J]. Exp Parasitol, 2000, 94: 269-272.
[22]	Foley DH, Harrison G, Murphy JR, et al. Mosquito bisection as a variable in estimates of PCR-derived malaria sporozoite rates[J]. Malar J, 2012, 11: 145.
[23]	张苍林, 郭祥瑞, 杨锐, 等. 2015年中缅边境盈江县疟疾疫点的调查与分析[J]. 中国寄生虫学与寄生虫病杂志, 2016, 34(5): 430-434.
[24]	王剑, 姜进勇, 郭晓芳, 等. 中国-老挝边境地区蚊虫群落结构和地理生态位特征分析[J]. 中国媒介生物学及控制杂志, 2017, 28(3): 209-215.
[25]	Sorchampa S, 郭晓芳, 王剑, 等. 老挝南塔省芒新县蚊虫种类调查[J]. 中国媒介生物学及控制杂志, 2017, 28(1): 66-68.
[26]	王剑, 董学书, 郭晓芳, 等. 老挝北部蚊虫种群组成及孳生习性调查[J]. 中国媒介生物学及控制杂志, 2016, 27(6): 549-554.
[27]	Al-Amin HM, Elahi R, Mohon AN, et al. Role of underappreciated vectors in malaria transmission in an endemic region of Bangladesh-India border[J]. Parasit Vectors, 2015, 8: 195-204.
[28]	St Laurent B, Burton TA, Zubaidah S, et al. Host attraction and biting behaviour of Anopheles mosquitoes in South Halmahera, Indonesia[J]. Malar J, 2017, 16: 310-319.
[29]	Lee HW, Shin EH, Cho SH, et al. Detection of vivax malaria sporozoites naturally infected in anopheline mosquitoes from endemic areas of northern parts of Gyeonggi-do (Province) in Korea[J]. Korean J Parasitol, 2002, 40: 75-81.
[30]	Oh SS, Hur MJ, Joo GS, et al. Malaria vector surveillance in Ganghwa-do, a malaria-endemic area in the Republic of Korea[J]. Korean J Parasitol, 2010, 48: 35-41.
[31]	Foley DH, Klein TA, Lee IY, et al. Mosquito species composition and Plasmodium vivax infection rates on Baengnyeong-do (island), Republic of Korea[J]. Korean J Parasitol, 2011, 49: 313-316.
[32]	Chang KS, Yoo DH, Ju YR, et al. Distribution of malaria vectors and incidence of vivax malaria at Korean army installations near the demilitarized zone, Republic of Korea[J]. Malar J, 2016, 15: 259.
[33]	Vythilingam I, Sidavong B, Chan ST, et al. Epidemiology of malaria in Attapeu Province, Lao PDR in relation to entomological parameters[J]. Trans R Soc Trop Med Hyg, 2005, 99: 833-839.
[34]	Trung HD, Van Bortel W, Sochantha T, et al. Malaria transmission and major malaria vectors in different geographical areas of Southeast Asia[J]. Trop Med Int Health, 2004, 9: 230-237.
[35]	周晓俊, 施文琦, 张仪, 等. 缅甸克钦地区媒介微小按蚊分布和传疟作用[J]. 中国病原生物学杂志, 2010, 5(8): 578-580, 584.
[36]	施文琦, 周晓俊, 张仪, 等. 中缅边境(西段)传疟媒介的初步调查[J]. 中国寄生虫学与寄生虫病杂志, 2011, 29(2): 134-137.
[37]	Pinontoan OR, Supadmanaba IGP, Manuaba IBA, et al. Local diversity and biting pattern of Anopheles species in Southern Minahasa[J]. Interdiscip Perspect Infect Dis, 2017, 2017: 1-6.