The effect of fumigation on the phosphine-susceptible and -resistant strains in the T. castaneum was evaluated using phosphine, ethyl formate, and combination with phosphine and ethyl formate (phosphine+ethyl formate). The Lethal Concentration Time (LCT)50 analysis of susceptible strains, late larva showed that phosphine (0.13 mg·h / L), ethyl formate (80.91 mg·h / L), and phosphine + ethyl formate (19.36 mg·h / L). The LCT50 of adult was 0.05 mg·h / L, 68.58 mg·h / L and 17.84 mg·h / L when treated with phosphine, ethyl formate, and phosphine + ethyl formate. The LCT50 of resistant strains, late larva was found to 33.32 mg·h / L of phosphine, 113.46 mg·h / L of ethyl formate and 129.85 mg·h / L of phosphine + ethyl formate, and the LCT50 of adult was 55.71 mg·h / L of phosphine, 85.39 mg·h / L, phosphine + ethyl formate 85.83 mg·h / L. The treatment of three fumigants (phosphine, ehtyl formate, and phosphine+ethyl formate) showed the possibility of controlling against T. castaneum of phosphine-susceptible and –resistant strains.

The fumigation activity of phosphine (PH3) to T. castaneum as a storage grain pest was evaluated. The lethal concentration time (LCT) value of each developmental stage (egg, early larva, late larva, pupa and adult) of T. castaneum was analyzed in 12 L desiccator. At the T. castaneum larva stages, exposure for 4 h showed low LCT value, especially in early stage larvae (LCT99 = 0.32 mg·h/L) which is very high susceptibility to PH3. However, T. castaneum eggs were observed very high tolerance to PH3 at LCT99 77.47 mg·h/L. Therefore, the fumigant activity of PH3 against T. castaneum can be found to be significantly different depending on developmental stage.

많은 산림해충들은 목재검역에서 문제가 되고 있어 포스핀(PH3)을 이용하여 광릉긴나무좀, 왕녹나무좀, 노랑애나무좀의 성충에 대한 살충 활성을 조사하였다. 3종의 나무좀류에 대하여 99% 살충활성을 보이는 LCT값은 각각 3.192 (광릉긴나무좀), 0.994 (왕녹나무좀), 0.501 mg ‧ h/L (노랑애나무좀) 순으로 나타났다. 포스핀을 처리한 3종의 나무좀류는 시간이 지남에 따라 살충활성도 증가하였다. 특히 광릉긴나무좀에서 는 0.4 mg/L 이상의 농도에서 약제처리 7일 후에 100%의 사충률을 보였다. 이러한 결과로 볼 때 포스핀이 3종의 목재해충 성충에 대해 메칠브 로마이드의 대체훈증제로 이용될 수 있을 것으로 판단된다.

Stored grain pests can cause reduction of grain quantity, quality, commercial value and germination rate. Susceptibility of three fumigants, methyl bromide, ethyl formate and phosphine, were assessed on Tribolium castaneum, which is an important stored grain pest. On susceptible insects, LCT50 of phosphine was 0.654mg h/L for egg, 0.127mg h/L for late larvae, 0.105mg h/L for pupae and 0.048mg h/L for adult stage, respectively. LCT50 of methyl bromide was 33.193mg h/L for egg, 14.585mg h/L for late larvae, 8.616mg h/L for pupae and 11.967mg h/L for adult stage, respectively. LCT50 of ethyl formate were 25.165mg h/L for egg, 80.912mg h/L for late larvae, 176.326mg h/L for pupae and 68.578mg h/L for adult stage, respectively. On resistant insects, LCT50 of phosphine were 82.325mg h/L for egg, 33.315mg h/L for late larvae, 73.546mg h/L for pupae and 55.707mg h/L for adult stage, respectively. LCT50 of methyl bromide were 19.250mg h/L for egg, 43.413mg h/L for late larvae, 76.842mg h/L for pupae and 19.387mg h/L for adult stage, respectively. LCT50 of ethyl formate were 87.552mg h/L for egg, 113.457mg h/L for late larvae, 200.122mg h/L for pupae and 85.394mg h/L for adult stage, respectively.

The susceptibility of three fumigants, methyl bromide, ethyl formate and phosphine, and concurrent treatment of ethyl formate and phosphine were tested on Lasioderma serricorne. Susceptibility assessment were performed by treating 5 to 6 initial dosage on every growth stages of L. serricorne. The LCT50 of methyl bromide was 13.896mg h/L for egg, 36.038mg h/L for late larvae, 25.172mg h/L for pupae and 21.758mg h/ L for adult, respectively. The LCT50 of phosphine was 0.317mg h/L for egg, 0.649mg h/L for late larvae, 3.748mg h/L for pupae and 0.703mg h/L for adult, respectively. In case of ethyl formate, the LCT50 was 43.657mg h/L for egg, 137.606mg h/L for late larvae, 72.676mg h/L for pupae and 52.951mg h/L for adult, respectively. Concurrent treatment of ethyl formate and phosphine was performed by treating 5 to 6 initial dosage of ethyl formate with 0.5 mg/L phosphine for 4 hours on every growth stages of L. serricorne. The LCT50 of ethyl formate concurrent treatment was 13.746mg h/L for egg, 8.156mg h/L for late larvae, 27.087mg h/L for pupae and 11.353mg h/L for adult, respectively, and these results indicate that concurrent treatment can control pest with lower dosage and shorter period. Sorption rates and ventilation periods of each fumigants were also calculated for efficacy and safety

Spotted wing drosophila, Drosophila suzukii, is an important pest of grape and strawberry because it lays egg inside fruit by damaging fruit surface. Recently, D. suzukii has been widely spreaded and causes several problems, so many countries include Australia designated D. suzukii as an important quarantine pest. Because of this, Korean farmers have trouble to export strawberry. In this study, we tested ethyl formate and phosphine to control D. suzukii, and also tested concurrent treatment of ethyl formate and phosphine to reduce phytotoxicity and enhance efficacy. When treated 35g/m3 of ethyl formate for 4 hours, mortality of egg, larvae, pupae and adult stages of D. suzukii was 22.2%, 21.1%, 19.2% and 28.3%, respectively. When treated 1g/m3 of phosphine for 24 hours, all stages of D. suzukii was completely controlled, but caused phytotoxic effect on strawberry. When treated with 35g/m3 of ethyl formate and 1g/m3 of phosphine concurrently for 4 hours, efficacy has been increased with less phytotoxicity than separate treatment. This result indicates that concurrent treatment enhanced efficacy with less phytotoxicity.

우드펠렛은 가정용, 상업용, 산업용 등으로 사용되며 특히 화력발전소에서 석탄 대용으로 수입량이 급증하여 연간 약 2,400만톤 가량 수입되고 있는 상황이다. 우드펠렛 검역훈증제로 기존에는 메틸브로마이드(이하 MB)가 최근까지 사용되어져 왔으나, MB의 높은 흡착률과 우드펠렛의 과도한 수용비로 인해 최종가스농도 기준에 미치지 못하여 재투약하는 사례가 빈번히 발생해 문제가 되어져왔다. 이를 해결하기 위해 농림축산검역본부에서는 올해 초 MB와 포스핀을 병행으로 처리하는 방식의 소독처리기준을 신설하였다. 새로운 소독처리기준이 설정됨에 따라 배기시 안전한 배기시간 수준을 설정하기 위해 병행처리 후 수착과 탈착되는 가스농도를 조사하였다. 실내 훈증상에 우드펠렛을 채우고 약제 처리한 결과 각 훈증제의 TLV(Threshold Limit Value)-TWA(Time Weighted Average) 기준이하로 농도가 감소되는 배기시간은 25℃일 때 MB는 120시간, 포스핀은 6시간으로 측정되었고 15℃일 때 MB는 186시간, 포스핀은 19시간으로 측정되었다.

This study investigated the fumigant activity of phosphine (PH3) on 2 kinds of mealybug (Pseudococcus longispinus and P. orchidicola (Hemiptera: Pseudococcidae)) adults and nymphs. All of the two mealybugs adults showed higher LCT99 values than nymphs, and P. longispinus showed higher tolerance than P. orchidicola in a 12 L desiccator. The absorption of phosphine on 13 nursery plants showed 12.2~41.5% difference depending on the plant. All of the mealybugs treated with phosphine 2 mg/L in 0.5 m3 fumigation chamber for 4 h showed 100% fumigant activity, except P. longispinus adult (approximately 90% at bottom part). However, when the exposure time was increased to 24 h, all of them showed 100% mortality. In the treatment of 10 m3 container, the 24 h treatment of phosphine showed 100% mortality to P. longispinus and P. orchidicola adults and nymphs. In all the experiments, no phytotoxicity of phosphine observed on 13 plants until 1 month after treatment.

수출입 묘목류에 있어서 깍지벌레류는 주요한 해충으로 다양한 묘목류에서 검출되고 있다. 깍지벌레류 중 우리나라의 소나무에 피해를 주고 있는 소나무굴깍지벌레는 연 2회 발생하는 해충으로 1세대는 4월, 2세대는 8월에 발생하여 소나무 잎에 피해를 준다. 따라서 본 연구에서는 수출입 묘목류에서 검출되는 깍지벌레류의 소독효과에 대한 정보를 제공하기 위하여 포스핀과 에틸포메이트 그리고 메틸브로마이드를 이용하여 소나무굴깍지벌레를 대상으로 훈증효과를 검정하였다. 포스핀을 소나무굴깍지벌레 알을 대상으로 0.06 mg/L 이상 처리시 난부화억제율은 100%였으며, 에틸포메이트 처리의 경우 성충은 2.6 mg/L, 알은 7.4 mg/L에서 100% 살충활성을 보였다. 그리고 메틸브로마이드를 40 g/㎥으로 2시간 훈증처리시 소나무굴깍지벌레 성충은 100% 치사하였으며, 알의 경우 26.0 mg/L을 처리했을 때 100%부화억제 효과를 보였다.

A combination using phosphine (PH3) and ethyl formate (EF) was performed to compensate for the disadventages (long exposure time and phytotoxicity) of a single substance. P. longisipinus was more susceptible to mixed phosphine and ethyl formate than P. orchidicola in 12 L dessicator. Mortality of mixed treatment was higher than aggregated mortalities that treated individually, so it indicated to have a synergic effect on each other. Ethyl formate was showed higher adsorption rate than phosphine on imported nursery plants and showed differences depending on the plant. P. longispinus and P. orchidicola was showed 100% mortality when phosphine and ethyl formate was treated as 1 g/m3 and 30 g/m3 for 4 hour in 0.5 m3 fumigation chamber and in 10 m3 container. Phytotoxicity was evaluated to mixture of phosphine and ethyl formate at 1 month after treatment.

Phosphine (PH3) resistance in the stored-products insect pests has been reported throughout the world in various insect species, including Rhyzopertha dominica, Tribolium castaneum, and Cryptolestes ferrugineus, leading farmers and fumigators to identify new fumigation tools to control PH3-resistant insect pests in storage facilities. Understanding PH3-resistance mechanisms in insects might contribute to providing clues for the development of new chemicals, including fumigants, to control various PH3-resistant insects. A proteomic study has shown 15 decreased proteins in the PH3-resistant R. dominica (CRD343 strain) in comparison to the PH3-susceptible R. dominica, and among those 15 proteins, dihydrolipoamide dehydrogenase (DLD), a protein involved in the Krebs cycle, was identified (Park et al., 2008). The DLD polymorphisms responsible for genetic resistance have disulfide active sites for PH3 binding and are highly sensitive to arsenic exposure after mutagenesis in insects (R. dominica and T. castaneum) and Caenorhabditis elegans (Schlipalius et al., 2012). Here, two PH3- resistant S. oryzae strains were used to understand the development of PH3 resistance in these insects. Acute toxicity test by PH3 on the two PH3-resistant strains was undertaken followed by ethyl formate inhibition study on cytochrome c oxidase activity. The Lineweaver-Burk plots after inhibition studies showed there were significantly difference in inhibition mode between the resistant strains and the control. The RT-qPCR analysis and the next-generation sequencing of the mitochondrial DNA revealed significant changes in metabolism and energy production. Taken together, the PH3 resistance in S. oryzae was definitely acquired by the overall transformation of biochemical reactions to overcome PH3 toxicity.

The respiration rate of PH3 susceptible strain was significantly higher than the resistant strain. The results showed no significant effect of oxygen level on the respiration rate of both strains. Phosphine reduced the respiration rate of both strains when it was applied in average concentrations. However, the rate of respiration of the resistant beetles increased significantly under a high level of phosphine. The increase of respiration rate was associated with the higher emission of VOCs which prove the acceleration of metabolic processes to face the phosphine action for survival. Flat grain beetle Cryptolestes pusillus and rusty grain beetle C. ferrugineus are similar insect species, but only C. ferrugineus is capable to develop a high phosphine resistance. A direct immersion solid phase microextraction gas chromatography-mass spectrometry (DI-SPME-GCMS) technology has identified the different fatty acids from PH3 resistant and susceptible strain of Tribolium castaneum.

알락하늘소(Anoplophora chinensis)와 유리알락하늘소(A. glabripennis)는 전 세계적으로 검역상 중요한 하늘소과 해충이다. 우리나라에서 이들은 자연 기주목과 분재에서 피해를 주고 있다. 본 연구에서는 분재에서 phosphine의 이들 두 하늘소에 대한 방제효과를 조사하였다. Phosphine은 솔수염하늘소 유충과 알에 대해 높은 훈증효과를 보였다. 2 g/m3 농도의 phosphine을 24시간 훈증처리 시 유리알락하늘소와 알락하늘소 난은 부화하지 못하였고, 알락하늘소 유충은 동일농도에 168시간 훈증처리 시 100% 치사되었다. 분재 생육기(2월)나 휴면기(1월) 모두에서 phosphine 훈증제는 2 g/m3 농도에서 두 알락하늘소를 100% 치사시켰다. 그러나 2월에 각 분재목[단풍나무(Acer palmatum)와 소사나무(Carpinus turczaninowii), 아그배나무(Malus sieboldii)]을 대상으로 훈증처리 시, 세 수종 모두에서 잎과 꽃잎의 크기가 작아지고, 꽃의 개화가 지연되는 약해가 발생되었다.

The current protocols of methyl bromide (MeBr) fumigation on imported nursery plants, which are seriously infested with quarantine mealy bugs and scales, could be reevaluated because there were no limited uses of loading ratio, packing types etc. Another emerging issues is that the workplace health issues on MeBr fumigation are related with poor ventilation etc as well as well-known phytotoxic damages on them. In previous report, we have evaluated the efficacies of ethyl formate (EF) and phosphine (PH3) on different types of mealybugs. Herein, the phytotoxic damages of EF and PH3 on Alocasia, Sansevieria, Cactaceae and Croton genera were evaluated in terms of chlorophyll contents, color changes etc. This study showed that EF and PH3 fumigation on imported plants could be applicable to replacement of MeBr with further confirmation trials.

Wood pellets are used by Biomass-solid refuse fuel (Bio-SRF) has been imported from many East-Asia countries especially,Vietnam in Korea. Wood pellets are made through grind and high-pressure process shows different physical characterizationlike high methyl bromide (MB) absorption unlike other wood materials. Furthermore, detected pests in wood pellets havesimilarity with detected pests in stored grain. In this study, we analyzed susceptibility on pupa and adult stages of redflour beetle, Tribolium castaneum in wood pellets against mixture of MB and phosphine (PH3). Mixture treatment ofMB and PH3 was more effective than single MB treatment compare with control. Pupal stage showed more tolerant thanadult stage on each fumigant and 100% mortality was determined at 27 mg/L of MB only at 20°C. However, PH3 increasedMB toxicity on pupae that 100% mortality was observed at 100:1 ratio of MB (5 mg/L) and PH3 (0.05 mg/L) at 20°C.Based on this study, mixture treatment of different type of fumigant will be applied to control of quarantine pests.

국내 정부양곡 보관창고에서는 알루미늄포스파이드 정제를 사용하여 쌀 해충 방제를 실시하고 있다. 이 약제는훈증시간이 4~7일로 장시간 소요되며, 제형 특성상 분말(잔사물)제거가 필요하고, 훈증완료 후에도 3~5%의 미반응물질이 남아있는 것으로 알려져 있다. 따라서 신속하고 안전한 새로운 훈증기술개발이 필요한 실정이며, Methylbromide(MB) 대체훈증제인 Ethyl formate(EF)와 Phosphine(PH3)의 병행처리를 통해 국내 주요 저곡해충인 가루좀벌레(장두)에 대한 약효를 조사하였다.가루좀벌레 성충을 대상으로 데시케이터 실험을 통해 도출된 100% 방제 가능 조건인 EF+PH3(16.6 + 1g/m3),24시간을 금속훈증상(1m3)에서 처리한 결과 가루좀벌레에 대해 100% 방제효과를 확인하였다. 본 연구는 양곡보관창고 훈증처리시 알루미늄포스파이드 대체제로서 EF+PH3의 사용 가능성을 제시하였다.

Fumigation activity of Frankliniella occidentalis, western flower thrips, and phytotoxicity were evaluated with phosphine(PH3) and ethyl formate (EF) in the asparagus. Fumigation activity of PH3 and EF showed in the order of egg (64.7 mg·h/Land 132.9 mg·h/L), nymph (5.5 mg·h/L and 14.8 mg·h/L), and adult (3.8 mg·h/L and 14.7 mg·h/L) respectively at 5˚C in12 L desiccator. The adsorption of PH3 was 24% at 5˚C. However, the adsorption of EF increased sharply to 98% at 5˚C.PH3 showed 100% control for all development stage of F. occidentalis with 4g/m3 in 0.65m3 at 2˚C for 48h exposure. Thephytotoxicity was not found. The results revealed that the EF is highly absorbable in asparagus, so it is not suitable as afumigant, but PH3 is considered to be suitable as an alternative MB fumigant for control of the F. occidentalis in asparagus.

The resistance of maize weevil(Sitophilus zeamais), an important agricultural pest of stored grain, on aluminium phosphide fumigant(PH3) has been reported recently in Korea. Many researchers reported that ethyl formate(EF), a potential methyl bromide alternative fumigant, showed good efficacy on several grain pests. In this trial, based on CTP (concentration × time products), we evaluated efficacy of EF on phosphine resistant and susceptible maize weevil. Each of insect strains were collected in grain storehouses and examined the PH3 resistance according to FAO method (Busvine, 1980). L(Ct)90 of PH3 resistant and susceptible strain was 48.3 and 0.3 g h m-3, respectively. When treated with EF, L(Ct)90 of PH3 resistant and susceptible strain was 90.0 and 110.9 g h m-3, respectively. In the case of EF, there was no difference between resistant and sucsecptible strain compare with PH3. These results indicates that EF is a potential alternative for aluminium phosphide resistance management in grain storehouses.

Worldwide, increasing numbers insecticide resistant insect is one of the main problem in agriculture not only in the field but also in the storage. The rusty grain beetle, Cryptolestes ferrugineus is one of the cosmopolitan insect that infests a wide range of stored cereals and related commodities. Until quite recently, phosphine (PH3) has been effective in controlling this species in worldwide including Korea. However, strongly resistant populations of RGB have been detected in Australia that could threaten market access of infested commodities. Resistant populations detected in Australia showed extremely high levels of resistance to phosphine, up to 1300 folds higher than that of susceptible strain. So here we tried to identify their phosphine resistance mechanism based on transcriptome analysis using RNaseq in adult stage. Over 10Gb were sequenced in each strains and some of specific P450 were over expressed in resistance strain.

The spotted wing drosophila (SWD), Drosophila suzukii, is a severe pest of berry fruits like strawberry, blueberry, sweet cherry and grape in terms of quarantine. For the disinfestation of SWD, methyl bromide (MeBr) is the only option in quarantine practice. However, MeBr's current use will be phased out due to chronic inhalation toxicity to fumigators and related worker as well as ozone depletion properties. To replace MeBr, we evaluated the efficacy of ethyl formate (EF) and phosphine gas (PH3) to all different developmental stages of SWD at practical fumigation condition (>25 ℃). To achieve complete control of all stages of SWD, PH3 required > 6 hr exposure when applied at > 1.0 g m-3. In case of 4hr EF fumigation, the LCt99 (LCt99, 99% lethal concentration × time) of EF to egg, larvae and pupae of SWD was 198.09, 77.42 and 115.55 g·h m-3, respectively.