Genetic diversity of Angiostrongylus cantonensis in Nan’ao Island of Guangdong Province

Abstract

Abstract:

Objective To investigate the genetic diversity of Angiostrongylus cantonensis in Nan’ao Island of Guangdong Province, and analyze the phylogenetic relationship among A. cantonensis isolates in different regions of China. Methods Three villages (Gongqian Village, Jinshan Village and Liudu Village) were selected in Nan’ao Island using the stratified random sampling method during 2015-2016. Rodents were captured by the night feeding and trapping method and were dissected to collect adult worms. Pomacea canaliculata and Achatina fulica collected from fields were examined for the stage-Ⅲ larvae, which were then used to infect SD rats to collect adult worms. Genomic DNA was extracted from the adult worms, and PCR was performed to amplify internal transcribed spacer 2 (ITS2) and cytochrome c oxidase subunitⅠ (COⅠ). The PCR products were sequenced and aligned by Clustal X1.83. The genetic diversity was analyzed with DnaSP5.10. The ITS2 sequences of A. cantonensis from different areas in the country, such as Wenzhou in Zhejiang (Accession number HQ540551.1), Shenzhen in Guangdong (Accession number HQ540546.1), Hualian in the Region of Taiwan (Accession number KF591125.1), were obtained from GenBank or published reports. The genetic distances between these strains were determined with Mega6.06. The phylogenetic trees were constructed by the neighbor-joining (NJ) and maximum parsimony (MP) methods to analyze the phylogenetic relationship. Results A total of 110 rodents were captured and 32 were positive for A. cantonensis (29.1%). And 1 190 P. canaliculata and 24 A. fulica were examined, with a larva infection rate of 6.1% (72/1 190) and 83.3% (20/24), respectively. The amplification products of ITS2 and CO I of A. cantonensis had a length of 693 bp and 1 174 bp, with a site mutation rate of 4.7% and 1.0%, respectively. The haplotype diversity index, nucleotide diversity index, and the average nucleotide difference index were 0.927, 0.007 and 4.549 respectively in ITS2, and 0.440, 0.004 and 3.743 in CO I. The genetic distance of A. cantonensis in Nan’ao Island with isolates from other sources was 0.181-0.775. Phylogenetic tree results showed that most isolates of A. cantonensis in Nan’ao Island were clustered into one branch with isolates from Shenzhen, Fuqing, Puer and Nanning, while a small portion of isolates from Nan’ao Island were clustered into an independent branch. Conclusions There exist some degrees of genetic differentiation and genetic diversity within the population of A. cantonensis in Nan’ao Island. They might have multiple types and sources, but more studies are needed to clarify this.

Key words: Angiostrongylus cantonensis, Internal transcribed spacer 2, Cytochrome c oxidase subunit Ⅰ, Genetic diversity, Phylogenetic relationship

CLC Number:

R383.19

Qiu-an HU, Shan LV, Yun-hai GUO, He-xiang LIU, Yi ZHANG. Genetic diversity of Angiostrongylus cantonensis in Nan’ao Island of Guangdong Province[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2017, 35(2): 130-135.

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References 23

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序列名称Sequence name	核苷酸组成/% Nucleotide composition/%				C	V	P	T_S/T_V	Hap	Hd	Pi	K	Tajima’s D
序列名称Sequence name	T	C	A	G	C	V	P	T_S/T_V	Hap	Hd	Pi	K	Tajima’s D
ITS2	32.1	17.4	27.0	23.5	634(95.3%)	31(4.7%)	15(2.3%)	3	21	0.927	0.007	4.549	-1.37（P > 0.05）
COⅠ	45.4	10.9	19.4	24.3	958(98.9%)	10(1.0%)	9(0.9%)	4	3	0.440	0.004	3.743	1.93（P > 0.05）

来源地 Area source	广东南澳 Guangdong Nan’ao	福建福清 Fujian Fuqing	广西南宁 Guangxi Naning	云南普洱 Yunnan Puer	海南琼中 Hainan Qiongzhong	浙江温州 Zhejiang Wenzhou	广东深圳 Guangdong Shenzhen	台湾花莲 Taiwan Hualian
广东南澳 Guangdong Nan’ao		0.022	0.022	0.022	0.022	0.023	0.022	0.075
福建福清 Fujian Fuqing	0.181		0.000	0.000	0.005	0.006	0.000	0.077
广西南宁 Guangxi Naning	0.181	0.000		0.000	0.005	0.006	0.000	0.077
云南普洱 Yunnan Puer	0.181	0.000	0.000		0.005	0.006	0.000	0.077
海南琼中 Hainan Qiongzhong	0.187	0.007	0.007	0.007		0.004	0.005	0.076
浙江温州 Zhejiang Wenzhou	0.192	0.011	0.011	0.011	0.004		0.006	0.077
广东深圳 Guangdong Shenzhen	0.181	0.000	0.000	0.000	0.007	0.011		0.077
台湾花莲 Taiwan Hualian	0.775	0.692	0.692	0.692	0.692	0.701	0.692