Imported 313 cases of Dengue fever and 16 cases of Zika virus disease were reported in the ROK during 2016. Aedesalbopictus has been reported as a major vector mosquito for Dengue and Zika virus. Various class of pesticides havebeen used locally different pesticide class. To investigate the pyrethroid and organophosphate pesticide resistance, genomicDNA was used for Allele-specific PCR (AS-PCR) genotyping of the acethylcholine esterase to detect Gly119Ser mutationsand kdr gene to detect Phe1534Cys mutations. Detoxification enzyme activities were assessed using Ae. albopictus fromYeosu and Jeju. Activities of four detoxification enzymes eg., glutathione S-transferase (GST), Non specific esterases (α-naphthylacetate and β-naphthyl acetate), and cytochrome C oxidase were determined for each Ae. albopictus strain. In our studies,Ae. albopictus showed only susceptible sequences for AchE and kdr. Activity of cytochrome P-450 and non specific esterasewas higher in a field population than a laboratory strain, except for GST. This study might be helpful to understandthe insecticidal susceptibility and resistant for effective vector mosquito control program.

Aedes albopictus is an important vector for yellow fever, dengue fever, chikungunya fever and Zika virus. This mosquitohas been exposed to organophosphates and pyrethroids in Republic of Korea for a long time. Using the direct contactmortality bioassay, susceptibility strain and two field populations of Ae. albopictus from Busan, and Damyang. Insecticidalproperty were indicated to RR ratio values (Resistant ratio to strain of susceptibility) of LC50 (Midian Lethal concentration)to organophosphats and pyrethroids. Genomic DNA was used for Allele-specific PCR (AS-PCR) genotyping of the acethylcholineesterase to detect Gly119Ser mutations and kdr gene to detect Phe1534Cys mutations. Detoxification enzyme activitiesof Ae. albopictus from Busan and Damyang were assessed using microplate enzyme activity assays. Activities of fourdetoxification enzymes eg., glutathione S-transferase(GST), Non specific esterases (α-naphthyl acetate and β-naphthyl acetate),and cytochrome C oxidase were determined for each Ae. albopictus strain. This study might suggest that Ae. albopictuscontrol programs should be prepared for the management of organophosphate and pyrethroid insecticide resistance.

The activities of three class, five acaricides (Permethrin, Deltamethrin, Fenitrothion, Chlorpyrifos, Imidacloprid) againstHaemaphysalis longicornis nymph, a major vector for Severe Fever with Thrombo cytopenia Syndrome (SFTS) and Lymedisease. The ticks distribute at vast open fields and various kind of habitats. Five acaricides were applied to ticks collectedform four areas(Gapyeong, Paju, Chuncheon, Wonju) to confirm the effect of acaricide. This areas was the area whereSFTS disease occurred and a large nember of ticks were collected. As a result, All areas is the most acaricidal effectto Deltamethrin. And two areas(Gapyeong, Paju) has the lowest acaricidal effect to Imidacloprid and two area(Chuncheon,Wonju) has the lowest acaricidal effect to Permethrin. Futher study will need to compare the acaricidal efficacy of theremaining seven province.

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.

The resistance levels to carbamate (CB) and organophosphate (OP) insecticides were determined by topical application in 14 field strains of Nilaparvata lugens. The resistance levels to CB and OP were 1.3~47.5-fold and 1.4~14.4-fold higher than a susceptible strain, respectively. A quantitative sequencing (QS) protocol was established to determine the allele frequencies of four acetylcholinesterase point mutations putatively associated with CB and OP resistance. The allele frequencies of the four mutations (G119A, F/Y330S, F331H and I332L) in field strains ranged from ca. 0.0~51.7%, 0.0~88.9%, 2.5~47.7%, 5.1~56.0% and 6.7~57.3%, respectively. The F331H and I332L were tightly linked each other, suggesting these mutations may occur simultaneously. In the correlation analysis, G119A was not well correlated with actual resistance levels (r2= < 0.232), whereas F331H and I332L showed a better correlation with the resistance levels of benzofuranyl methylcarbamates (r2= 0.595). This finding indicates that F331H and I332L mutation frequencies may be used as molecular markers for detecting carbamate resistance in N. lugens. A QS protocol detecting the F331H and I332L mutation frequencies would be employed as a supportive tool for rapid monitoring of CB insecticide resistance levels in N. lugens

The resistance levels against carbamates (CB) and organophosphates (OP) were determined through bioassay and quantitative sequencing (QS) methods in 16 field populations of Nilaparvata lugens. The resistance levels to CB and OP by bioassay were 1.3~47.5-fold and 1.4~14.4-fold higher than a susceptible strain, respectively. The QS protocol was established to determine the allele frequencies of eight point mutations on acetylcholinesterase putatively associated CB and OP resistance. The allele frequencies of four mutations in local populations (G119A, F/S330Y, F331H and I332L) ranged from ca. 0.0~51.7%, 1.0~44.3%, 8.5~57.3% and 7.12~56.6%, respectively. The average prediction limits were –9.6±5.1~7.7±2.9%. The F330Y, F331H and I332L were tightly linked each other, suggesting these mutations may occur simultaneously. In the correlation analysis, G119A was not well correlated with both insecticides (r2= less 0.25), whereas F/S330Y, F331H and I332L showed better correlation with the resistance levels of carbamate (r2=0.590) than organophosphate (r2=0.235). This finding indicates that F/S330Y, F331H and I332L mutation frequencies are suitable for detecting carbamate resistance in N. lugens. QS will be applicable for the rapid monitoring of resistance levels to CB insecticides in N. lugens.

A quantitative sequencing (QS) protocol was established for predicting the frequencies of the A298S and G324A mutations in the diamondback moth (Plutella xylostella) type-1 acetylcholinesterase locus, putatively involved with organophosphate and carbamate insecticideresistance. The nucleotide signal ratio at each mutation site was generated from sequencing chromatograms and plotted against the corresponding resistance allele frequency. Frequency prediction equations were generatedfrom the plots by linear regression, and the signal ratios were shown to highly correlated with resistance allele frequencies (r2>0.987). QS analysis of 15 representative regional field populations of DBM in Korea revealed that the allele frequencies of both A298S and G324A were over 70% in most field populations. As judged by inhibition assay, all populations showed resistance to paraoxon, DDVP, carbaryl, and carbofuran. In addition, different DBM strains exhibited differential sensitivities to both OPs and CBs depending on the structure of inhibitor, implying that the resistance of DBM against OPs and CBs is saturated and widespread in Korea.

Organophosphate pesticides have their inhibitory action on the chorinesterase. It is likely that application of organophosphate pesticides invoke retardation in activity as well as death of arthropods. This study is aimed to find changes in foraging activity of ants due to application of fenitrothion, one of most widely used organophosphate pesticides. Foraging activity (FA) of ants was observed by using bait cards at pesticide sprayed pine stand and unsprayed stand before and after aerial application of fenitrothion in 2003 and 2004. FA of Paratrechina flavipes which was too abundant to be statistically compared, significantly decreased from 2 hours to 10 days after application of the pesticide but recovered at 16 day in 2003 and partially recovered at 18 day and completely at 31 day in 2004. FA of other ant species also significantly decreased during the retardation period of FA of P. flavipes. Ratio of ants feeding on baits significantly decreased during the retardation period of FA. However, decrease of interspecific inhibition on feeding action, which was expected for negligence on other ants caused by the chorinesterase inhibition was not observed.

Organophosphate pesticides were extracted with 70% acetone and then transferred to dichloromethane. Extracts were applied to open-column chromatography with florisil The final extract was analyzed by gas chromatography with nitrogen-phosphorus detector(GC/NPD). Recoveries of the 18 organophosphate pesticides were ranged from 88.7% to 100.0% for the narrow-bore capillary GC(Ultra-2). The minimum detectable level of this analytical method was 0.019 - 0.035 ㎎/㎏. Sample throughput(extraction, open-column chromatography, and GC analysis) was decreased considerably (8h per sample).