Emergence of resistant two-spotted spider mite (TSSM) can induce the over usage of standard amount of acaricides and result in various side effects. Rapid resistance monitoring is essential step for the efficient management of resistant populations by enabling the selection of appropriate acaricides. Here, we evaluated the 19 acaricides to determine its suitability for residual contact vial bioassay (RCV) by using PyriF strain as a reference. Twelve acaricides (Amitraz Abamectin, Bifenthrin, Bifenazate, Chlorfenapyr, Cyenopyrafen, Cyflumetofen, Endosulfan, Fenothiocarb, Monocrotophos, Omethoate and Tebufenpyrad) revealed the dose-dependent mortality within 8 h, whereas other remaining acaricides (Dicofol, Etoxazole, Fenbutatin oxide, Fenpyroxymate, Flufenoxuron, Spiromesifen and Pyridaben) did not. This finding suggests that the application of RCV method is limited depending on the mode of action and physicochemical properties of each acaricide. Resistance levels to 12 acaricides were determined for four field populations of TSSM by using RCV diagnostic kit. All TSSM populations showed the highest sensitivity to cyflumetofen, indicating that it would be most effective in controling field populations. RCV diagnostic kit would enable to provide crucial information for choosing the most appropriate acaricides in the field.

Emergence of resistant two-spotted spider mite (TSSM) can induce the over usage of standard amount of acaricides and result in various side effects. Rapid resistance monitoring is essential step for the efficient management of resistant populations by enabling the selection of appropriate acaricides. Here, we evaluated the 10 acaricides to determine its suitability on residual contact vial bioassay (RCV) by using PyriF strain as standard. Five acaricides (Abamectin, monocrotophos, tebufenpyrad, bifenazate and cyflumetofen) revealed the dose-dependent mortality within 8 h, whereas other remaining acaricides (Fenbutatin oxide, fenpyroxymate, flufenoxuron, spiromesifen and etoxazole) did not. This finding suggests that the application of RCV method is limited depending on the mode of action and physicochemical properties of each acaricide. Resistance levels to five acaricides (Abamectin, monocrotophos, tebufenpyrad, bifenazate and cyflumetofen) were determined for four field populations of TSSM by using RCV diagnostic kit. All TSSM populations showed the highest sensitivity to cyflumetofen, indicating that it would be most effective in controling field populations. RCV diagnostic kit would enable to provide crucial information for choosing the most appropriate acaricides in the field.

고분자물질의 사용으로 인한 건축 재료의 다변화에 따라 화재 시 발생되는 연소가스 또한 다양해지고 있다. 그중 HBr은 NES 713과 BS 6853, FTP Code Part 2에서 연소 가스의 독성 평가 시 측정되는 독성가스이다. 특히 산업안전물질보건자료(MSDS)에서는 HBr가스를 흡입할 경우 화상을 일으키며 호흡부전, 두통 등을 유발시킬 수 있고 50ppm의 HBr가스에 노출된 사람은 생명과 건강에 즉시 영향을 받을 수 있는 매우 유독한 연소가스이다. 본 논문에서는 HBr 표준가스에 노출된 동물의 운동성 측정실험인 가스유해성 시험 및 HBr가스에 노출된 마우스의 생물학적 검사 결과를 비교 분석하여 HBr의 연소 독성에 관한 연구를 수행하였다.

Termites are a major problem for home and business owners around the world. Termites gnaw through wood and burrow under the foundations of buildings causing a great deal of damage to the structure of the building. Phosphine (PH3) is very effective fumigant and is widely used to control pests. PH3 rapidly penetrate through treated material, so it replaced many conventional fumigants for treatment of durable materials. In this study, we have investigated the effectiveness of PH3 fumigation on wood pests, Reticulitermes speratus kyushuensis. We evaluated two bioassay methods; wooden cube (10×10×10 cm) and insect breeding dish in comparison with effectiveness of PH3 penetrations into the timber block. Fumigation to adults of R. speratus was carried in a desiccator system for 24 h at 5 and 15℃. As a result, LC99 of PH3 to R. speratus in wooden cubes and insect breeding dish at 5℃ was 0.183 and 0.177 mg L-1, respectively. LC99 of PH3 in wooden cubes and insect breeding dish at 15℃ was 0.077 and 0.078 mg L-1, respectively. There were no significant differences between the two bioassay methods. Thess results indicate that PH3 described herein merit further study as potential fumigants for termite control.

Methyl bromide (MB) is a widely used fumigant in most of the countries for quarantine purpose. However, MB has been phasing out and under control in many countries because it is listed as an ozone-depleting substance under the Montreal Protocol. In this study, we have investigated the effectiveness of phosphine fumigation on wood pests for developing an MB alternatives. We evaluated two bioassay methods; wooden cube (10×10×10 cm) and normal fumigation procedures in comparison with effectiveness of phosphine (PH3) penetrations into the timber block. Fumigation to adults of Reticulitermes speratus was carried in a desiccator system for 24hr at 5 and 15 ℃. As a result, LC99 of PH3 to R. speratus in wooden cubes and insect breeding dish at 5℃ was 0.183 and 0.177 mg L-1, respectively. LC99 of PH3 in wooden cubes and insect breeding dish at 15℃ was 0.077 and 0.078 mg L-1, respectively.

The two-spotted spider mite Tetranychus urticae is a worldwide crop pest with a high insecticide resistance and an extensive host range. The aim of the present study was to evaluate the effect of PaeciPora®, which was formulated from the aerial conidia of an entomopathogenic fungus Paecilomyces lilacinus strain HY-4, to control T. urticae in cucumber field. In the field study, conidia of P. lilacinus HY-4 and a chemical acaricide azocyclotin were investigated for their control of the adult females of T. urticae. The strain produced a mortality of 56.0% on day 3 and 63.6% on day 7 post-treatment respectively at 1×107 conidia/mL, and no evidence of a mortality benefit was seen in the control group. Additionally, in the pesticide injury test, no agrochemical damage was found in hot pepper, watermelon, Chinese cabbage, oriental melon or strawberry by spraying PaeciPora® on them. The results indicated the possibility of the use of P. lilacinus HY-4 as a microbiological control agent against T. urticae in the Integrated Pest Management program.

The green peach aphids, Myzus persicae (Sulzer), feed consistently and regularly throughout all stages of nymphs and adult. That is, honey dew production (HDP), which indicates a state of feeding, occurs at regular intervals within a given stage, and moving (MV), which interrupts feeding, tends to occur scarcely. We used these behaviors to test whether the sound stimuli acted as a stress in this species. We conducted the playback experiment in which acoustic stimuli with a combination of five frequency (100, 500, 1000, 5000, and 10000 Hz) and three intensity levels (66, 78, 90 dB SPL) were presented to all stages of aphids. Under all acoustic stimuli, HDP occurred less often, and MV occurred more often and longer. The results of two-way multivariate ANOVA showed that the stimulus frequency was a significant factor for variation in aphid behaviors in the playback experiment. However, there was no intensity effect on aphid’s behavior. Based on the electrical penetration graph experiment, aphid feeding was frequently stopped and lasted only for short durations under acoustic stimuli. The results of both experiments demonstrated that the acoustic stimuli were effective in inducing feeding suppression in this species. Thus, the acoustic stimuli may be a stressor in the green peach aphid. Furthermore, the behavioral bioassay employed in this study may be used to assess the effect of various stressors on aphids.

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.

Two point mutations (V419L and L925I) in the voltage-sensitive sodium channel (VSSC) α-subunit gene have been identified in deltamethrin-resistant bedbugs. To predict resistance allele frequencies of sodium channel mutations (V419L and L925I) in bedbugs at a population level, we developed quantitative sequencing (QS) protocol. The signal ratios between resistant and susceptible nucleotides were generated from sequencing chromatogram and plotted against the corresponding resistance allele frequencies. Linear regression coefficients of the plots were close to 1 (r2 = 0.9928 and 0.9998), suggesting that the signal ratios are reliable correlated with the resistance allele frequencies. To enable on-site monitoring of pyrethroid resistance in bed bugs, residual contact vial (RCV) bioassay method was established and used to determine median lethal concentration (LC50) values to deltamethrin for various bed bug strains. Resistance allele frequencies in these bedbug strains predicted by QS were correlated well with the RCV bioassay results, confirming the roles of two mutations in pyrethroid resistance. Taken together, employment of QS in conjunction with RCV bioassay should greatly facilitate resistance monitoring of bedbugs in the field.

The stable fly, Stomoxys calcitrans L., is an important pest of livestock. Stable flies are considered as mechanical vectors of veterinary disease. Pyrethroids and organophosphates have been widely used for stable fly control. To establish resistance monitoring molecular tool, we isolated the partial cDNA and genomic fragments of voltage-sensitive sodium channel and acetylcholinesterase genes encompassing the well known conserved sites for resistance-associated mutations. To examine the current status of stable fly resistance to pyrethroids and organophosphates mediated by the nerve insensitivity mechanism in Korean population of S. calcitrans, DNA-based genotyping in conjunction with residual contact vial (RCV) bioassay were conducted with 11 representative regional field populations. No resistance-associated mutations were detected in these S. calcitrans populations, suggesting that these populations are likely still susceptible to both pyrethroids and organophosphates. Establishment of RCV bioassay protocol and availalbility of target site sequence information will greatly facilitate resistance monitoring of S. calcitrans in the field.

Varroa destructor is an ecto-parasite mite and worldwide pest of the honey bee Apis mellifera L. The pyrethroid tau-fluvalinate (Apistan), an acaricide that is tolerated by honey bees, has been used for varroa mite control since the mid 1980s. Even though various resistances to tau-fluvalinate in varroa mites have been reported from Europe, Israel, and USA, the nature of tau-fluvalinate resistance in varroa mites in Korea has never been investigated. To investigate and understand tau-fluvalinate resistance in varroa mites in Korea, we conducted bioassay in several apiaries located different regions in Korea. To understand the molecular mechanisms underlying the difference of tau-fluvalinate resistances in varroa mites, partial genomic DNA fragments of a voltage-sensitive sodium channel gene from varroa mites were cloned and sequenced, since tau-fluvalinate is known to act on the sodium channels directly. Two novel mutations in sodium channels of varroa mites were present in eight apiaries. Two mutations might be a geographical polymorphism of sodium channel of varroa mites in Korea.

To establish a rapid diagnosis method for the monitoring of acaricide resistance in the two-spotted spider mite, Tetranychus urticae, we evaluated the performance of residual contact vial (RCV) method as a routine bioassay for T. urticae by using two widely used acaricides, abamectin and tebupenpyrad. Appropriate concentrations of test acaricides were dissolved in acetone and evenly coated (100 μl) onto the inside wall of a 4-ml glass vial using a rolling machine. The average survival times in untreated control vial was longer than 12 hrs in the absence of food or water regardless of cap being closed or open. Webbing behavior of mites inside the vial, which may interfere with maximum chemical contact, began to be observed from 8 hrs post treatment. The minimum concentrations causing 100% mortality within 8 hrs posttreatment in a susceptible strain of T. urticae were determined to be 30 and 60 ppm in abamectin and tebupenpyrad, respectively. Dose-response curve was significantly affected by temperature in both acaricides, in which the knockdown rate increased greatly as temperature increased. The endpoint mortality at 6-8 hrs posttreatment, however, was not significantly affected by temperature. Nymphal stage of mites showed more rapid intoxication response than adults but endpoint mortality at 6-8 hrs posttreatment was not substantially different between developmental stages. When compared with the results from conventional spray method, RCV method showed moderate to high correlation coefficients (r=0.51~0.98), suggesting that it is a reliable in determining susceptibility of T. urticae. The vial-coated pesticides were stable at least one year when stored at -20°C as determined by LC-MS/MS analysis. Moreover, there was no significant difference in the bioassay results in repeated experiments with three different persons, indicative of high reproducibility of RCV. The RCV diagnostic kit, when used by farmers on site, should provide crucial and essential information for the selection of most suitable acaricides for different field populations of T. urticae.

Bioassay of mosquito, Culex pipiens molestus, larva was investigated by several environment-friendly insecticides. These insecticides were Novaluron as chitin synthesis inhibitor, Metho-xyfenozide as ecdysone agonist, Pyriproxyfen as juvenile hormone mimic, and Spiromesifen as lipid biosynthesis inhibitor. The 50% lethal concentration (LC₅₀) of these insecticides were 0.00039, 0.07193, 0.65006 and 0.04839 ppm, respectively. Novaluron has lower concentration than any other insecticide. To determine the treatment time against larval stages, insecticides were applied to different larval stages of C. pipiens molestus. Mortality ratios of mosquito larva treated with Novaluron were 100.0, 84.5, 71.0 and 48.5% on 2, 4, 7 and 10 days after hatching from eggs, respectively. Otherwise, with the other insecticides, mortality ratios were under 80% with 2 days old larva. When exposure periods were tested to 3 or 4 days old larva against 4 insecticides, at least 3 hours were needed to 100% control effect against Novaluron, and over 12 hours with other insecticides.

Three sex pheromone components (cis-8-dodecenyl acetate (Z12Ac), trans-8-dodecenyl acetate (E12Ac), cis-8-dodecenol (Z12OH)) of the Oriental fruit moth, Grapholita molesta, were chemically synthesized. Especially to increase the composition of cis-stereoisomer, a triple bond intermediate was hydrogenated at -20℃ with catalytic Pd/BaSO₄. The resulting product consisted of the acetates with a stereoisomer ratio in 92:8 (Z:E). The biological activity of the synthesized pheromone compounds was analyzed both in male responses and orientation disruption. The indoor pheromone effect was determined by male flight behavior showing wing movement in response to lure. Different mixtures of the synthetic pheromone components were prepared by mixing acetate and alcohol components in 100:0, 99:1, and 90:10 (g/g) and tested with a comparison of a standard commercial pheromone lure. The highest pheromone effect was observed in only acetate mixture (100:0) and the effect was reduced with the addition of the alcohol component. This indoor pheromone effect could be observed in field monitoring trial, in which 100:0 mixture showed the highest trap catches. Orientation disruption assay was conducted indoor by using a cage, in which the center had a commercial lure on sticky plate and the four candidates were placed at 6 ㎝ away from the central lure on each of four directions. Test males were released to the arena during overnight (12 h) and then the caught males on the sticky plate were counted. The synthesized pheromone as well as the commercial pheromone showed 100% orientation disruption. However, the orientation disruption effect was reduced with decrease in the number of the surrounding disrupting pheromone baits. These results clearly suggest that the synthesized sex pheromone of G. molesta is biologically active and can be used for field mating disruption.