Development of a multiplex PCR assay for the identification of eight species members of the Thai Hyrcanus Group (Diptera: Culicidae)

Hempolchom, Chayanit; Otsuka, Yasushi; Baimai, Visut; Thongsahuan, Sorawat; Saeung, Atiporn; Taai, Kritsana; Srisuka, Wichai; Somboon, Pradya; Choochote, Wej

doi:10.1007/s13355-013-0207-1

Cited by 17 publications

(19 citation statements)

References 32 publications

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“…The DNA of An. minimus groups and the Hyrcanus Group were extracted from legs or wings of each mosquito for further species confirmation [ 33 , 34 ].…”

Section: Methodsmentioning

confidence: 99%

Receptivity to malaria in the China–Myanmar border in Yingjiang County, Yunnan Province, China

Chen

Zhang

Zhou

et al. 2017

Malar J

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BackgroundThe re-establishment of malaria has become an important public health issue in and out of China, and receptivity to this disease is key to its re-emergence. Yingjiang is one of the few counties with locally acquired malaria cases in the China–Myanmar border in China. This study aimed to understand receptivity to malaria in Yingjiang County, China, from June to October 2016.MethodsLight-traps were employed to capture the mosquitoes in 17 villages in eight towns which were categorized into four elevation levels: level 1, 0–599 m; level 2, 600–1199 m; level 3, 1200–1799 m; and level 4, > 1800 m. Species richness, diversity, dominance and evenness were used to picture the community structure. Similarity in species composition was compared between different elevation levels. Data of seasonal abundance of mosquitoes, human biting rate, density of light-trap-captured adult mosquitoes and larvae, parous rate, and height distribution (density) of Anopheles minimus and Anopheles sinensis were collected in two towns (Na Bang and Ping Yuan) each month from June to October, 2016.ResultsOver the study period, 10,053 Anopheles mosquitoes were collected from the eight towns, and 15 Anopheles species were identified, the most-common of which were An. sinensis (75.4%), Anopheles kunmingensis (15.6%), and An. minimus (3.5%). Anopheles minimus was the major malaria vector in low-elevation areas (< 600 m, i.e., Na Bang town), and An. sinensis in medium-elevation areas (600–1200 m, i.e., Ping Yuan town). In Na Bang, the peak human-biting rate of An. minimus at the inner and outer sites of the village occurred in June and August 2016, with 5/bait/night and 15/bait/night, respectively. In Ping Yuan, the peak human-biting rate of An. sinensis was in August, with 9/bait/night at the inner site and 21/bait/night at the outer site. The two towns exhibited seasonal abundance with high density of the two adult vectors: The peak density of An. minimus was in June and that of An. sinensis was in August. Meanwhile, the peak larval density of An. minimus was in July, but that of An. sinensis decreased during the investigation season; the slightly acidic water suited the growth of these vectors. The parous rates of An. sinensis and An. minimus were 90.46 and 93.33%, respectively.ConclusionsThe Anopheles community was spread across different elevation levels. Its structure was complex and stable during the entire epidemic season in low-elevation areas at the border. The high human-biting rates, adult and larval densities, and parous rates of the two Anopheles vectors reveal an exceedingly high receptivity to malaria in the China–Myanmar border in Yingjiang County.

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“…The DNA of An. minimus groups and the Hyrcanus Group were extracted from legs or wings of each mosquito for further species confirmation [ 33 , 34 ].…”

Section: Methodsmentioning

confidence: 99%

Receptivity to malaria in the China–Myanmar border in Yingjiang County, Yunnan Province, China

Chen

Zhang

Zhou

et al. 2017

Malar J

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“…Then, specimens were preserved in microtubes at − 20 °C and brought back to the laboratory. Anopheles mosquitoes belonging to the Hyrcanus Group were sent to the UMR-HSM Laboratory at the Institut de Recherche pour le Développement, Montpellier, France, for molecular species identification using allele-specific multiplex PCR assays [23].…”

Section: Mosquito Collections and Species Identificationmentioning

confidence: 99%

“…argyropus and An. sinensis) [23,24]. Seven of these species are widely distributed throughout the country, while An.…”

Section: Introductionmentioning

confidence: 99%

“…barbirostris group, which also belongs to the Myzorhynchus Series. Mosquitoes of the two groups share some similar morphological characteristics such as a humeral crossvein composed of a remigium patch with dark scales and narrow apical pale bands on the midtarsi, comparable wing patterns, narrow apical fringe spot on the wings, narrow tarsal bands, and highly variable hindtarsal banding patterns [23]. These characteristics, as well as loss or damage of mosquito samples during field activities, are the leading causes of misidentification that may skew data on the current distribution of Anopheles vectors across the country and ultimately reduce the benefits of vector control approaches.…”

Section: Introductionmentioning

confidence: 99%

“…The ITS2 region is relatively short in length (generally less than 1 kb) and highly conserved, which are suitable criteria for the design of universal primers allowing its fast amplification and sequencing [32]. Based on ITS2 polymorphisms, Hempolchom et al [23] have designed specific primer sequences, which were successfully used in a single-round multiplex PCR for the identification of Anopheles species within the Hyrcanus Group.…”

Section: Introductionmentioning

confidence: 99%

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Species diversity and insecticide resistance within the Anopheles hyrcanus group in Ubon Ratchathani Province, Thailand

et al. 2020

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Background Members of the Anopheles hyrcanus group have been incriminated as important malaria vectors. This study aims to identify the species and explore the insecticide susceptibility profile within the Anopheles hyrcanus group in Ubon Ratchathani Province, northeastern Thailand where increasing numbers of malaria cases were reported in 2014. Methods Between 2013 and 2015, five rounds of mosquito collections were conducted using human landing and cattle bait techniques during both the rainy and dry seasons. Anopheles mosquitoes were morphologically identified and their insecticide susceptibility status was investigated. Synergist bioassays were carried out with An. hyrcanus (s.l.) due to their resistance to all insecticides. An ITS2-PCR assay was conducted to identify to species the Hyrcanus group specimens. Results Out of 10,361 Anopheles females collected, representing 18 taxa in 2 subgenera, 71.8% were morphologically identified as belonging to the Hyrcanus Group (subgenus Anopheles), followed by An. barbirostris group (7.9%), An. nivipes (6.5%), An. philippinensis (5.9%) and the other 14 Anopheles species. Specimens of the Hyrcanus Group were more prevalent during the rainy season and were found to be highly zoophilic. Anopheles hyrcanus (s.l.) was active throughout the night, with an early peak of activity between 18:00 h and 21:00 h. ITS2-PCR assay conducted on 603 DNA samples from specimens within the Hyrcanus Group showed the presence of five sisters species. Anopheles peditaeniatus was the most abundant species (90.5%, n = 546), followed by An. nitidus (4.5%, n = 27), An. nigerrimus (4.3%, n = 26), An. argyropus (0.5%, n = 3), and An. sinensis (0.2%, n = 1). All An. hyrcanus (s.l.) specimens that were found resistant to insecticides (deltamethrin 0.05%, permethrin 0.75% and DDT 4% and synergist tests) belonged to An. peditaeniatus. The degree of resistance in An. peditaeniatus to each of these three insecticides was approximately 50%. Addition of PBO (Piperonyl butoxide), but not DEF (S.S.S-tributyl phosphotritioate), seemed to restore susceptibility, indicating a potential role of oxidases as a detoxifying enzyme resistance mechanism. Conclusions A better understanding of mosquito diversity related to host preference, biting activity and insecticide resistance status will facilitate the implementation of locally adapted vector control strategies.

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Scanning electron microscopy of Anopheles hyrcanus group (Diptera: Culicidae) eggs in Thailand and an ultrastructural key for species identification

et al. 2013

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The eggs of Anopheles argyropus, Anopheles crawfordi, Anopheles nigerrimus, Anopheles nitidus, Anopheles paraliae, Anopheles peditaeniatus, Anopheles pursati, and Anopheles sinensis are described with the aid of scanning electron micrographs. Comparisons of the egg structure among the eight species showed that the eggs differed with respect to the following characteristics: the deck-complete (An. argyropus, An. nigerrimus, An. paraliae, An. peditaeniatus, and An. sinensis); variable (complete, split and incomplete decks found together within an egg batch/An. crawfordi); and division into an area at each end (An. nitidus and An. pursati). The ratios of the entire length per maximal deck width within the area covered by floats were 3.33-6.86 (An. sinensis), 8.78-18.20 (An. peditaeniatus), 13.67-22 (An. nigerrimus), 26.33-44.25 (An. paraliae), and 26.99-75.94 (An. argyropus). The numbers of float ribs were 21-27 (An. peditaeniatus) and 28-34 (An. nigerrimus), and the total numbers of anterior and posterior tubercles were 6-8 (An. paraliae) and 9-11 (An. argyropus). Exochorionic sculpturing was of reticulum type (An. argyropus, An. crawfordi, An. nigerrimus, An. nitidus, An. paraliae, An. peditaeniatus, and An. sinensis) and pure tubercle type (An. pursati). Attempts are proposed to construct a robust key for species identification based on the morphometrics and ultrastructures of eggs under scanning electron microscopy.

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Development of a multiplex PCR assay for the identification of eight species members of the Thai Hyrcanus Group (Diptera: Culicidae)

Cited by 17 publications

References 32 publications

Receptivity to malaria in the China–Myanmar border in Yingjiang County, Yunnan Province, China

Receptivity to malaria in the China–Myanmar border in Yingjiang County, Yunnan Province, China

Species diversity and insecticide resistance within the Anopheles hyrcanus group in Ubon Ratchathani Province, Thailand

Scanning electron microscopy of Anopheles hyrcanus group (Diptera: Culicidae) eggs in Thailand and an ultrastructural key for species identification

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