Objective To identify the common species of sandflies in Haikou city based on morphological characteristics, and determine the species specific molecular identity. Methods Sandflies were collected overnight by light-trap monthly in the pigsty, chicken or duck coops or human dwellings around village and urban-rural fringe areas in Haikou city during August 2015 to July 2016, as well as in Sansha city in August, 2017. After counting, partial sandflies were randomly selected and classified by morphological characteristics. Then the genomic DNA was extracted from identified specimens, the mtDNA Cytb gene fragment was sequenced and analyzed to determine the species-specific molecular identity. The sandflies collected from Sansha city were identified by molecular marker, and the taxonomic status was discussed. Results A total of 377 sandflies samples were collected in Haikou city, which belonged to 4 species in 2 genera, as Sergentomyia bailyi Sinton, S.iyengari Sinton, S. squamipleuris Newstead, and Phleboto musstantoni Newstead. The sandflies adults were active throughout the year with the peak of density in June and November. The intraspecific genetic distance of mtDNA Cytb gene sequences among the 4 species was from 0 to 0.046. The range of the interspecific distance among the 9 species (including 5 species mtDNA Cytb gene sequences downloaded from NCBI database) was from 0.112 (P. wui with P. chinensis) to 0.219 (P. stantoni and P. caucasicus). All sandflies specimen collected from Sansha city were S. bailyi by molecular identity. The most genetic distance was 0.049 between Haikou and Sansha population. Conclusion Sandflies were active throughout the year in Haikou city, including two peaks of density. The identity of mtDNA Cytb gene sequences of 4 common sandflies in Hainan province was identified. There was obvious genetic divergence in S. bailyi populations.
[1] Depaquit J. Molecular systematics applied to Phlebotomine sandflies:review and perspectives[J]. Infect Genet Evol, 2014,28:744-756.
[2] Service MW. Medical entomology for students[M]. 3rd ed. Liverpool:Cambridge University Press,2004:90-98.
[3] 熊光华,金长发,管立人. 中国的白蛉[M]. 北京:科学出版社,2016:15-25.
[4] 杨曼尼,马雅军. 我国白蛉的分类现状[J]. 国际医学寄生虫病杂志,2008,35(1):46-49.
[5] Zhang LM, Leng YJ. Eighty-year research of Phlebotomine sandflies (Diptera:Psychodidae) in China (1915-1995). Ⅱ. Phlebotomine vectors of Leishmaniasis in China[J]. Parasite, 1997,4(4):299-306.
[6] 管立人. 中国白蛉(双翅目:毛蛉科)调查研究工作的展望[J]. 中国寄生虫学与寄生虫病杂志,2013,31(4):310-314.
[7] 冷延家. 在海南岛继续发现的蛉种与新种白蛉:方亮白蛉Phlebotomus fanglianensis sp. nov.的记述[J]. 昆虫学报, 1964,13(1):118-128.
[8] 杨明磊,李逸明,陈辉莹,等. 我国西沙群岛吸血昆虫的种群动态和鼠形动物病原感染检测[J]. 第二军医大学学报, 2016,37(3):355-359.
[9] Kumar NP, Srinivasan R, Jambulingam P. DNA barcoding for identification of sand flies (Diptera:Psychodidae) in India[J]. Mol Ecol Resour,2012,12(3):414-420.
[10] de Souza Pinto I, das Chagas BD, Rodrigues AAF, et al. DNA barcoding of neotropical sand flies (Diptera, Psychodidae, Phlebotominae):species identification and discovery within Brazil[J]. PLoS One,2015,10(10):e0140636.
[11] Polseela R, Jaturas N, Thanwisai A, et al. Towards monitoring the sandflies (Diptera:Psychodidae) of Thailand:DNA barcoding the sandflies of Wihan Cave, Uttaradit[J]. Mitochondrial DNA A DNA Mapp Seq Anal,2016,27(5):3795-3801.
[12] Kocher A, Gantier JC, Gaborit P, et al. Vector soup:high-throughput identification of Neotropical phlebotomine sand flies using metabarcoding[J]. Mol Ecol Resour,2017,17(2):172-182.
[13] Dokianakis E, Tsirigotakis N, Christodoulou V, et al. DNA sequencing confirms PCR-RFLP identification of wild caught Larroussius sand flies from crete and cyprus[J]. Acta Trop, 2016,164:314-320.
[14] Gajapathy K, Tharmasegaram T, Eswaramohan T, et al. DNA barcoding of Sri Lankan phlebotomine sand flies using cytochrome c oxidase subunitⅠ reveals the presence of cryptic species[J]. Acta Trop,2016,161:1-7.
[15] Esseghir S, Ready PD, Ben-Ismail R. Speciation of Phlebotomus sandflies of the subgenus Larroussius coincided with the late Miocene-Pliocene aridification of the Mediterranean subregion[J]. Biol J Linn Soc,2000,70(2):189-219.
[16] 冷廷家. 贝氏白蛉及其亚种分类的研究[J]. 暨南大学学报:自然科学版,1980,1(2):23-30.
[17] 高景鹏,李凯利,陈辉莹,等. 菊酯类杀虫剂对海口市白纹伊蚊的效果测定[J]. 中国热带医学,2017,17(9):867-870.
[18] Zhang L, Ma YJ, Xu JN. Genetic differentiation between sandfly populations of Phlebotomus chinensis and Phlebotomus sichuanensis (Diptera:Psychodidae) in China inferred by microsatellites[J]. Parasit Vectors,2013,6(1):115.
[19] Zhang L, Ma YJ. Identification of Phlebotomus chinensis (Diptera:Psychodidae) inferred by morphological characters and molecular markers[J]. Entomotaxonomia,2012,34(1):71-80.
[20] Parvizi P, Naddaf SR, Alaeenovin E. Molecular typing and phylogenetic analysis of some species belonging to Phlebotomus (Larroussius) and Phlebotomus (Adlerius) subgenera (Diptera:psychodidae) from two locations in Iran[J]. Iran J Arthropod Borne Dis,2010,4(2):1-10.
[21] Krüger A, Strüven L, Post RJ, et al. The sandflies (Diptera:Psychodidae, Phlebotominae) in military camps in northern Afghanistan (2007-2009), as identified by morphology and DNA "barcoding"[J]. Ann Trop Med Parasitol,2011,105(2):163-176.
[22] Berdjane-Brouk Z, Koné AK, Djimdé AA, et al. First detection of Leishmania major DNA in Sergentomyia (Spelaeomyia) darlingi from cutaneous leishmaniasis foci in Mali[J]. PLoS One,2012,7(1):e28266.
[23] Gajapathy K, Peiris LB, Goodacre SL, et al. Molecular identification of potential leishmaniasis vector species within the Phlebotomus (Euphlebotomus) argentipes species complex in Sri Lanka[J]. Parasit Vectors,2013,6(1):302.
[24] Kasap OE,Vot'pka J,Alten B. The distribution of the Phlebotomus major complex (Diptera:Psychodidae) in Turkey[J]. Acta Trop,2013,127(3):204-211.
[25] Bounamous A, Lehrter V, Hadj-Henni L, et al. Limits of a rapid identification of common Mediterranean sandflies using polymerase chain reaction-restriction fragment length polymorphism[J]. Mem Inst Oswaldo Cruz,2014,109(4):466-472.
[26] Depaquit J,Randrianambinintsoa FJ,Jaouadi K,et al. Molecular and morphological systematics of the sandfly Sergentomyia (Sintonius) clydei Sinton, 1928 and questions about its record in the Seychelles[J]. Infect Genet Evol,2014,21:41-53.
[27] Ebrahimi S,Bordbar A, Parvizi P. Genetic dynamics in the sand fly (Diptera:Psychodidae) nuclear and mitochondrial genotypes:evidence for vector adaptation at the border of Iran with Iraq[J]. Parasit Vectors,2016,9(1):319.
[28] 张丽,马雅军. 基于线粒体DNA的我国中华白蛉群体遗传分化研究[J]. 中国热带医学,2016,16(10):947-952.
[29] Yogeswari S,Srinivasan R. A note on variations in morphological features of the Phlebotomine sand fly Sergentomyia bailyi (Diptera:Psychodidae) in a population from Pondicherry UT, India[J]. J Med Entomol,2016,53(3):712-716.
[30] Tharmatha T, Gajapathy K, Ramasamy R, et al. Morphological and molecular identification of cryptic species in the Sergentomyia bailyi (Sinton,1931) complex in Sri Lanka[J]. Bull Entomol Res,2017,107(1):58-65.
