海南省玛瑙螺物种鉴定及其感染广州管圆线虫的调查  

Species identification of Achatina and investigation of its infection with Angiostrongylus cantonensis in Hainan province

作　　者：高素真 舒畅 吕鑫 焦惠生 周云飞 吕志跃 GAO Suzhen;SHU Chang;LÜXin;JIAO Huisheng;ZHOU Yunfei;LÜZhiyue(School of Public Health,Hainan Medical University,Haikou,Hainan 571199,China;School of Basic Medicine and Life Science,Hainan Medical University,Haikou,Hainan 571199,China;Department of Parasitology,Zhongshan School of Medicine,Sun Yat-sen University,Guangzhou,Guangdong 510080,China)

机构地区：[1]海南医学院公共卫生学院,海南海口571199 [2]海南医学院基础医学与生命科学学院,海南海口571199 [3]中山大学中山医学院寄生虫学教研室,广东广州510080

出　　处：《热带医学杂志》2024年第4期480-485,489,I0002,共8页Journal of Tropical Medicine

基　　金：海南省重大科技计划项目(ZDKJ202003,ZDKJ2021035);海南省重点研发计划项目(ZDYF2020120);海南省院士创新平台项目(YSPTZX202133);国家自然科学基金(82072303,81572023,81371836);国家寄生虫资源库(NPRC-2019-194-30);广东省科技计划项目(2019B030316025);广东省自然科学基金(2019A1515011541);国家重点研发项目(2021YFC2300800,2021YFC23008001,2021YFC23008002,2021YFC2300003)。

摘　　要：目的结合野外调查和实验室分析,了解海南现存玛瑙螺物种多样性及其携带广州管圆线虫的情况。方法采用分层随机抽样的调查方法,于2022年12月-2023年10月在海南省18个市县53个采样点采集玛瑙螺样本1060份,根据形态特征对样本进行初步鉴定,并取形态特征有差异的部分样本进行DNA提取和COI基因扩增,通过GenBank数据库内序列比对确定物种,并使用Mega 11软件构建进化树。同时采用肺检法和消化法了解其感染广州管圆线虫的情况并分析影响因素。结果在采集的1060份玛瑙螺样本中,通过形态学鉴定和分子鉴定确定海南存在两种玛瑙螺,其中褐云玛瑙螺占70.57%(748/1060),虎纹玛瑙螺占29.43%(312/1060)。形态学区分主要在胼胝部颜色上;分子鉴定显示,与褐云玛瑙螺处于同一支的样本,COI基因序列一致性在99.84%~100.00%;采集的未定种样本与虎纹玛瑙螺在同一进化分支上,COI基因一致性为90.75%~95.97%。二者与亲缘关系最近的物种间平均遗传距离分别为0.000~0.003和0.043~0.102。海南省玛瑙螺整体广州管圆线虫感染率为5.85%。褐云玛瑙螺和虎纹玛瑙螺的广州管圆线虫感染率分别为5.08%(38/748)和7.69%(24/312),差异无统计学意义(χ^(2)=2.728,P>0.05);二者感染度中位数分别为107.70条/g和37.32条/g,差异有统计学意义(Z=2.067,P<0.05)。海南省内各市县玛瑙螺总体感染率最高为澄迈20.63%(13/63)、乐东16.67%(12/72)和海口11.11%(4/36);褐云玛瑙螺感染率最高为澄迈25.00%(9/36)、乐东16.36%(9/55)和陵水16.00%(4/25);虎纹玛瑙螺感染率最高为乐东、海口、澄迈和万宁,分别为17.65%(3/17)、16.67%(3/18)和14.81%(4/27)。市县间感染率在玛瑙螺全部样本和褐云玛瑙螺样本内差异均有统计学意义(χ^(2)=68.33、37.16,P均<0.05),在虎纹玛瑙螺样本内差异无统计学意义(χ^(2)=3.578,P>0.05)。结论存在未定种玛瑙螺,高度疑似虎纹玛瑙螺,为岛内入侵物种玛瑙Objective To investigate the species diversity of Achatina and the infections of Angiostrongylus cantonensis in Hainan province by field investigation and laboratory analysis.Methods A total of 1060 samples were collected from 53 sampling sites in 18 cities and counties in Hainan province from 2022-2023,and the samples were preliminarily identified according to their morphological characteristics.DNA extraction and COI gene amplification were performed in some samples with different morphological characteristics,and the species was identified by sequence alignment in GenBank database.The phylogenetic tree was constructed using Mega 11 software.At the same time,lung examination and digestion methods were used to investigate the infection of A.cantonensis,and the influencing factors were analyzed.Results Among the 1060 samples,two species of Achatina were identified in Hainan province by morphological and molecular identification,with Achatina fulica accounting for 70.57%(748/1060)and A.panthera accounting for 29.43%(312/1060).Morphological differentiation was mainly based on the color of callosal region.The sequence identity of COI gene was 99.84%-100.00%in the samples belonging to the same clade as A.fulica.The COI gene identity was 90.75%-95.97%between the unidentified species samples and A.panthera.The average genetic distances between the two species and the closest related species were 0-0.003 and 0.043-0.102,respectively.The overall A.cantonensis infection rate was 5.85%in Achatina sp.in Hainan province.The prevalence of A.cantonensis infection was 5.08%(38/748)in A.fulica and 7.69%(24/312)in A.panthera,and the difference was not statistically significant(χ^(2)=2.728,P>0.05).The median infection intensity of the two groups were 107.70 bars/g and 37.32 bars/g,respectively,and the difference was statistically significant(Z=2.067,P<0.05).The highest infection rates of all snails were found in Chengmai(20.63%,13/63),Ledong(16.67%,12/72)and Haikou(11.11%,4/36).The highest infection rates of A.fulica were found i

关 键 词：玛瑙螺 COI基因 广州管圆线虫 

分 类 号：Q958[生物学—动物学] R383.1[医药卫生—医学寄生虫学]