말라리아는 Anopheles 모기들에 의해 전파되며, 대한민국에서는 코로나 팬데믹 기간 동안(2020~2022) 약 200~300명대의 말라리아 환자가 발생하였으나, 지난 2023년에는 그 수가 폭증해 약 800명의 환자가 발생하였다. 현재까지 모기를 방제하기 위한 가장 효율적인 수단은 살충제를 사용한 방제이나, 지속적으로 이러한 화합물에 노출된 모기 개체군은 살충제에 저항성을 갖게 되는 것으로 알려져 있다. 이에 본 연구에서는 대한민국의 주요 말라리아 발생 지역인 비무장지대 부근 및 이외 지역인 용산, 평택, 오산에서 채집을 진행하였으며, 채집된 An. sinensis에 대해서 살충제 저항성과 관련되어 있다고 알려진 유전자인 acetylcholinesterase-1(ace-1)와 voltage-gated sodium channel(vgsc) 영역에 대한 저항성 돌연변이 보유 여부를 각각 확인하였다. 실험 결과 채집된 모든 지역에서 G119S(ace-1), L1014F,C(vgsc) 돌연변이가 발견되었으며, 그 빈도는 계절과 장소에 따라 차이를 나타내었다. 본 실험 결과는 향후 말라리아 감염 억제를 위한 매개체 연구에 활용될 수 있을 것으로 기대된다.
The distribution of mosquitoes collected in Cheongju area in 2016 and 2017 were compared. The collection sites wereestablished from residential area, migratory bird sanctuary, and cattle feedlot using BL trap and BG trap. Culex pipienswere the most collected in residential area, and Aedes vexans were the most collected in migratory bird sanctuary andcattle feedlot in both years. BG trap was more collected than BL trap in residential area and migratory bird sanctuary.However, cattle feedlot was more collected in BL trap. A variety of mosquito species were collected from BG trap thanBL trap. However, the number of Anopheles sinensis collected in cattle feedlot showed significant differences betweenBL trap and BG trap. Based on the results of this study, it is considered that effective monitoring of mosquitoes mightbe done by using appropriate trap type according to the region
Anopheles sinensis is an important vector for Plasmodium vivax and thus has been targeted with pyrethroids in Republic of Korea. Using the direct contact mortality bioassay, two field populations of An. sinensis from Ganghwa-gun and Goyang-si were characterized for their resistance to pyrethroids with RR ratio values (Resistant ratio to imidacloprid) of 125.6 to 203.8 folds and 80.0 to 120 folds, respectively. Genomic DNA was used for Allele-specific PCR (AS-PCR) genotyping of the sodium channel genes to detect L1014S mutations. The homozygous susceptible Leu/Leu genotype in Ganghwa-gun and Goyang-si was 5.0% and 11.3% and the resistance genotypes were 95.0% and 88.7%, respectively. The homozygous Phe/Phe resistance genotype was the most prevalent as 35.4% in Ganghwa-gun and 44.0% in Goyang-si. Hence, this study suggests that malaria vector control programs should be prepared for the management of pyrethroid insecticide resistance.
Anopheles sinensis is an important vector for Plasmodium vivax and thus has been targeted with organophosphates in Republic of Korea. Using the direct contact mortality bioassay, two field populations of An. sinensis from Paju-si and Yeoncheon-gun were characterized for their resistance to organophosphats with RR ratio values (Resistant ratio to imidacloprid) of 446.9 to 756.3 folds and 502.1 to 861.7 folds, respectively. Genomic DNA was used for Allele-specific PCR (AS-PCR) genotyping of the acethylcholine esterase to detect Gly119Ser mutations. The homozygous susceptible Ser/Ser genotype in Paju-si and Yeoncheon-gun was 20.0% and 20.9% and the resistance genotypes were 80.0% and 79.1%, respectively. The homozygous Gly/Ser resistance genotype was the most prevalent as 60.7% in Paju-si and 41.9% in Yeoncheon-gun. Hence, this study suggests that malaria vector control programs should be prepared for the management of organophosphate insecticide resistance.
Anopheles sinensis is an important vector for Plasmodium vivax and thus has been targeted with pyrethroids in Republic of Korea. Using the direct contact mortality bioassay, two field populations of An. sinensis from Paju-si and Yeoncheon-gun were characterized for their resistance to pyrethroids with RR ratio values (Resistant ratio to imidacloprid) of 96.8 to 167.2 folds and 34.2 to 98.4 folds, respectively. Genomic DNA was used for Allele-specific PCR (AS-PCR) genotyping of the sodium channel genes to detect L1014S mutations. The homozygous susceptible Leu/Leu genotype in Paju-si and Yeoncheon-gun was 3.3% and 37.1% and the resistance genotypes were 96.7% and 62.9%, respectively. The homozygous Phe/Phe resistance genotype was the most prevalent as 45.0% in Paju-si and 31.5% in Yeoncheon-gun. Hence, this study suggests that malaria vector control programs should be prepared for the management of pyrethroid insecticide resistance.
Control of the vector mosquitoes is critical for implementation of patient’s decrease. Vector mosquito control has been mainly carried out using pesticides in Republic of Korea (ROK), which has developed the resistance against selected insecticides. Voltage-gated sodium channel (VGSC) and Acethylcholine esterase (AchE) are the target of several classes of insecticides. Genetic mutations of VGSC and AchE have been documented to confer resistances to pyrethroids and organophosphates in mosquitoes, respectively. In our studies, the insecticidal resistable level of An. sinensis from four malaria risk areas near Demilitarized Zone (DMZ) was measured. The partial sequence of An. sinensis voltage gated sodium channel (VGSC) IIS6 domain for Kdr gene and ace1 gene for AchE was sequenced. An. sinensis from two areas demonstrated significantly higher resistance to pyrethroids and mutation rates of Kdr gene were comparatively higher. In all tested areas An. sinensis showed high insecticidal resistance to organophosphates and > 70% of mutation rates of ace1 gene ware higher. Understanding the mechanisms of insecticide resistance among vector mosquitoes populations is very important to efficient and effective vector control.
The insecticidal activities of materialsderived from Dryopteris crassirhizoma Nakai against third instar larvae of three species mosquitoes (Culex pipiens, Aedes albopictus and Anopheles sinnensis) were evaluated using a direct contact mortality bioassay.
The methanol extracts of D. crassirhizoma showed 100%, 87.8% and 100% larvicidal activity at 1,000 ppm against Cx. pipiens, Ae. albopictus and An. sinensis, respectively. Hexane fraction showed 100% larvicidal activity three species mosquitoes at 500 ppm after 24 hrs. Purification of the biologically active constituents from the hexane extraction with larvicidal activity was done using silica gel column chromatography. H1 and H3 fractions gave 100% mortality to Cx. pipiens, Ae. albopictus and An. sinensis at 100 ppm. H1 fraction separated with methanol to give a H111 through centrifugation. Fraction of the biologically active constituents from the H3 fraction with larvicidal activity was done using methanol. H31 fraction was determined 100% mortality to Cx. Pipiens, Ae. albopictus and An. sinensis at 50 ppm, respectively. Two fractions were analyzed C14H22O (H111, MW206.0) and C11H14O4 (H31, MW210.08) by GC and GC-MS. D. crassirhizoma derived compounds described herein could be useful for managing filed populations as larvicide of Cx. pipiens, Ae. albopictus and An. sinensis.
An assessment was made of the toxicity of 12 insecticides, three essential oils and Bacillus thuringiensis var. israelensis (Bti) alone or in combination with the oil major constituents, (E)-anethole (AN), (E)-cinnamaldehyde (CA) and eugenol (EU), to third instars from bamboo forest collected Aedes albopictus and rice paddy field collected Anopheles sinensis resistant to various groups of insecticides. The toxicity of the test insecticides, essential oils and binary mixtures of Bti and the oil constituents (1:1 ratio) was evaluated using a direct-contact mortality bioassay. Binary mixtures of B.t.i. and CA, AN or EU were significantly more toxic against Ae. albopictus larvae (LC50, 0.0084, 0.0134 and 0.0237 mg/l) and An. sinensis larvae (0.0159, 0.0388 and 0.0541 mg/l) than either B.t.i. (1.7884 and 2.1681 mg/l) or CA (11.46 and 19.43 mg/l), AN (16.66 and 25.11 mg/l) or EU (24.60 and 32.14 mg/l) alone. Based on the co-toxicity coefficient (CC) and synergistic factor (SF), the three binary mixtures operated in a synergy pattern (CC, 140.7–368.3 and SF, 76–213 for Ae. albopictus CC, 75.1–245.3 and SF, 40–136 for An. sinensis).The binary mixtures of Bti and essential oil constituents described, particularly (E)-cinnamaldehyde, merit further study as potential mosquito larvicides for the control of malaria vector mosquito populations in light of global efforts to reduce the level of highly toxic synthetic insecticides in the aquatic environment.
Field-collected populations of mayflies, Ephemera orientalis were tested for susceptibility to 10 different insecticides using a direct-contact mortality bioassay. Ephemera orientalis subimagoes were susceptible to the insecticides chlorpyrifos, fenitrothion and chlorfenapyr with LD50 values of 69.7, 78.8 and 81.9μg/♀, and adults had LD50 values of 71.9, 78.8 and 85.4μg/♀, respectively. Susceptibility ratios(SRs) of subimagoes and adults of E. orientalis to the 10 insecticides were 1.0 to1.2 folds. The mayflies showed higher susceptibility to organophosphates than to pyrethroids. The SRs of Anopheles sinensis to E. orientalis were 514 to 1,438 fold higher for organophosphates (LD50 values of 0.05 to 0.23μg/♀) and 62 to 1,155 fold higher forpyrethroids (LD50 values of 0.13 to 2.41μg/♀). The SRs of Culex pipiens to E. orientalis were 606 to 3,595 fold higher for organophosphates with LD50 values of 0.02-0.17μg/♀ and 81 to 1,365 fold higher for pyrethroids with LD50 values of 0.11- 1.83μg/♀. These results indicate that the use of ineffective insecticides will result in unsatisfactory control against field populations of the subimagoes and adults of E. orientalis. This work was supported by a research grant from the National Vector Control and Surveillance work performed by the Korean National Institute of Health.
Bacillus thuringiensis는 포자형성기 동안에 위생해충이나 농업해충에 독성을 보이는 내독소 단백질을 생성한다. 내독소 단백질의 모기 유충 방제효과를 높이기 위해, 광합성에 관여하는 psbA promoter로 모기 살충성 cry11Aa유전자를 발현하는 pSyn4D벡터를 제작하고, 모기 유충이 먹이로 이용하는 Synechocystis PCC6803에 형질 전환시켰다. 형질 전환체들은 kanamycin이 포함된 배지에서 선발되었으며, 정상적인 생물검정을 통해 형질 전환체 Tr2C를 선발하였다. cry11Aa 유전자는 형질전환체의 genomic DNA에 안정적으로 결합되어 있는 것을 PCR을 이용하여 확인하였다. 형질전환체 Tr2C는 약 72-kDa크기의 Cry11Aa 단백질을 발현하였으며, 작은빨간집모기(Culex tritaeniorhynchus) 3령 유충과 중국얼룩날개모기(Anopheles sinensis) 3령 유충에 75%가 넘는 살충력을 보였다. 모기 유충에 대한 형질전환체의 반수치사시간(LT)은 작은빨간집모기 유충과 중국얼룩날개모기 유충에 대해 각각 2.1일과 0.7일이었다. 이상의 결과들은 형질전환체 Tr2C가 모기 유충방제에 유용하게 이용될 수 있음을 보여준다.