In order to industrialization of the entomopathogenic nematode (EPN) isolated in Korea, it is necessary to switch to infective juveniles. Since the conversion to infective juveniles in the end of solid culture is important against the outside environment, it has a great influence on insecticidal efficacy and shelf life. We investigated whether the concentration of CO2 and NH3 at the end of the solid culture was artificially increased to affect the infective juvenile induction rate. Optimal exposure time for conversion to infective juveniles was 5 minutes for CO2 and 3 minutes for NH3. Optimal treatment time was 9 days after EPN inoculation for both CO2 and NH3. And optimum concentration was 80~90% for both CO2 and NH3 for EPN infective juveniles stage conversion.

In order to develop solid mass cultivation technology that maximizes insecticide of entomopathogenic nematode(EPN) isolated from Korea, we have studied the optimization of solid culture medium and production conditions. The optimized conditions from the harvest yields and the insecticidal activity against the Galleria mellonella larvae were yeast extract 2.0%, soybean flour 20%, whole milk powder 3%, olive oil 5% and egg yolk 5%. It was also found that the optimum condition of the carrier(polyurethane) content was about 8-12%, the culture time of symbiotic bacteria was about 48 hours, and the inoculation concentration of nematode was 4,000-5,000 per g medium. And the optimal harvesting point in solid culture conditions was confirmed after 12 days of inoculation.

Insensitive acetylcholinesterase (AChE) was determined to be basically involved in a EPN-resistant (ER) strain and ER contaminated susceptible (CS) strain of diamondback moth (DBM, Plutella xylostella L.), as estimated by the AChE inhibition assay using DDVP as a inhibitor in nondenaturing electrophoresis gel. ER strain exhibited very high insensitivity revel, high resistance ratio and two point mutations (G227A, A201S) in ace1. CS strain showed intermediate insensitivity level, low resistance ratio and a point mutation (G227A). This finding suggests that the A201S mutation along with reported G227A mutation (Baek et al, 2005) can be main player to develop the organophosphate resistance. Additionally, surveyed seven local population DBMs saturated G227A mutation and A201S mutation mixed in some population. Also A441G mutation was detected in some population, but there was no significant correlation. These results suggest that A201S mutation could be one of the good candidate for molecular diagnosis marker of resistance monitoring.

Insensitive acetylcholinesterase (AChE) was determined to be basically involved in a EPN-resistant (ER) strain of diamondback moth (DBM, Plutella xylostella L.), as estimated by the AChE inhibition assay using DDVP and thiodicarb as inhibitors in nondenaturing electrophoresis gel. AChEs were clearly separated into four different bands (a major band and three minor bands) in susceptible strain (CS) and all bands inhibited by used inhibitors almost same level, however, only two bands (a major band and a minor band) showed in that of ER strain and major band showed higher insensitivity. Moreover, ER strain showed cross resistance against used inhibitors and DDVP highly inhibited esterase in both strains. About 2kb of ace1,2 cloned and point mutations were detected in ER strain.

In order to evaluate the degradation organophosphorus pesticide, EPN, in water environment, the effects of water temp.(10℃, 30℃), pH(3-11) and sunlight on its degradation were investigated during 10 days. The degradation rate of EPN(200 rpm) was faster at higher water temp. and higher pH, i.e., its degradation rate at pH 3, 5, 7, 9, 11 was 57, 63, 66, 69, 75%(10℃), and 70, 74, 79, 91, 97%(30℃) after 10 days, respectively. The effect of water temp. on its degradation was little in acidic condition, but was rather great in alkaline condition, with time. EPN was degraded fast at the alkaline condition by photolysis. At the condition of pH 11, EPN was degraded fast at the early stage in the first 2 days, but after that the degradation rate was weakened.

To study the residue of organophosphorus pesticides in the ocean environment on the Cheju island, EPN[O-ethyl-O-4-nitrophenyl phenylphosphonothioate] and monocrotophos [Dimethyl-1-methyl-2-methy] carbamoylvinylphosphate] of organophosphorus pesticides are surveyed on coastal environment of Cheju city and Wimi in May and October 1996. The qualified limit detection of EPN and monocrotophos are 0.005ng/mL and 0.006ng/mL., in the water by GC-FPD, respectively. The qualified limit detection of EPN and monocrotophos are 0.010ng/g and 0.012ng/g in the solid such as sediment and sea organism by GC-FPD, respectively. EPN and monocrotophos are not detected in seawater and sediment. Moreover EPN and monocrotophos are neither detected in seaweed cava (Ecklonia cava), Agar (Gelidium amansii), turban sell(Batillus cornutus) and sea urchin(Anthocidaris Crassispina). EPN, monocrotophos used in the farm on the Cheju island are not residued in the coastal environment in the Cheju island.