Methyl bromide (MB) fumigation is still allowed to use in quarantine treatment on imported timbers and logs, even though IPPC(International Plant Protection Convention) was strongly recommended to reduce their use in case of existing alternatives. A newly developing MB alternative fumigant, ethanedinitrile (EDN), showed the potential in terms of better efficacy to various timber pests and penetration into the timber than MB. Herein, for suggestion of quarantine guideline in Korea, we had verified the efficacies under different temperature conditions(5, 5-15, >15℃) and monitored TLV(Threshold limited value) of EDN post-fumigation for worker safety. The 6 scheduled doses of EDN depending on types of timber pest and temperature conditions had evaluated with PVC-Tarp tent(>100m3) with 40-50% loading factors of imported logs. To control normal wood related pests such as Japanese termite (Reticulitermes speratus) and bark beetle(Cryphalus fulvus), the scheduled doses 30, 40 and 40 g/m3 for 24hr fumigations showed >99% efficacy at 5, 5-15 and >15℃, respectively. The scheduled doses 100, 120 and 150 g/m3 for 24hr fumigations was also successful to gain the proved efficacies at 5, 5-15, >15℃, respectively, for the quarantine guideline to notorious wood related pests such as Japanese pine sawyer (Monochamus alternatus) and pine wilt nematode (Bursaphelenchus xylophilus). The recommended ventilation times with atmospheric conditions at ports was > 1 and > 2 hr under the fully uncovered and partially uncovered of fumigation tent conditions, respectively.

The German cockroach, Blatella germanica, is one of the most important household pest that can cause disease on human. Conventional contact insecticides show less efficacy because it is hard to be contacted or digested by cockroach that lives inside small crevice. Ethyl formate(EF), which is a potential fumigant to control various insect pests such as stored grain insects and quarantine pests, can be penetrated wherever air exist and shows toxicity on respiratory system of insect. In this trials, we've demonstrated practical and commercial scale fumigations of EF on German cockroach. Efficacy of EF was described in terms of concentration × time (CT) product. At practical fumigation, EF gave 100% control of German cockroach nymph and adult when the CT product was 43.2g h m-3. When 600m3 scale warehouse fumigation for 15 hours with 5g m-3 of EF, nymph and adult stage of German cockroach were completely controlled. The ventilation time to reduce EF concentration under TLV(Threshold Limit Value, EF=100ppm) was less than 1 hour. These results show that EF can be applied at warehouse or mill house to control German cockroach effectively.

With the increasing trend of global trades and protection of agro-ecosystem in importing and exporting countries, quarantine and pre-shipment(QPS) fumigation for perishable commodities are critical in terms of quality maintenance of fumigated commodities as well as eradicating quarantine pest. There are limited use of methyl bromide(MB) fumigation on fruits and vegetables due to not only worker safety issue at low temperature condition but phytotoxic damage to valuable commodities. We are here presented that commercial development of methyl bromide (MB) alternatives such as ethyl formate and phosphine gas formulated with carbon dioxide, that are increasing consumer and worker safety and avoiding phytotoxic damage in routine MB fumigation.

Methyl Bromide (MB) has been regulated by montreal protocol due to ozone depletion since 1992, and there is a need to develop MB alternative fumigation methods. To find out the efficacies and tolerance in developmental stages of citrus mealybugs (Planococcus citri) which frequently infested in imported pineapples, phosphine(PH3) gas was tested in different exposure times (2~24 hr) and concentrations (0.1∼2g/m3) in small desiccators at 8℃. Based on Lethal 99% concentration × time products (LCTP 99%) of PH3 gas to target developmental insect, 2g/m3 PH3 was applied for 24 hr in 300 m3 fumigation facility, which was designed in well sealed and evacuation system (5 air exchange/min). We monitored PH3 gas in fumigation facility to confirm CTP and checked PH3 gas in atmospheres in air ventiliation process after fumigations to make quarantine guideline and protocol for worker safety as well as efficacy to target pest.

Methyl bromide(MB) is a fumigant for quarantine and pre-shipment(QPS) that can eradicate target pests within short fumigation periods. However, MB, an ozone-depleting substance, is scheduled to be phased out in the second decade of this century. In contrast to MB alternatives for soil fumigation, there is no ideal alternative fumigant to MB for QPS of perishable commodities. It is important that all target insect pests are killed within a short fumigation time. The citrus mealybug(Planococcuscitri), a problematicquarantine pest from Korea, was exposed to ethyl formate and phosphine for different times and to different concentrations of EF(0.0∼33.4 mg/L) and PH3(0.25∼1.0 mg/L) in small desiccators. The eggs showed more tolerance than the nymphs and adults. To control the eggs, they were treated with a mixture of EF and PH3 for 2 and 4-hr. The egg stage showed 100% mortality with EF:PH3(25.1:0.25, 16.7:0.5, and 16.7:1.0 mg/L) at 4-hr. Treatment with a mixture of EF and PH3 for 4-hr at 8°C can control all stages of the citrus mealybug.

Aphids are common pests frequently found in imported and exported fruits and vegetables. Methyl bromide(MB), a Quarantine and Pre Shipment(QPS) fumigant, could offer eradication of aphids within short period. However, MB is limited in use because of poor gas evaporation at low temperature(<5℃) and there is phytotoxic effect or damage on quality in post-harvest vegetables and fruits even at >5℃. Two candidates of MB alternative, ethyl formate(EF) and phosphine(PH3), are used and being investig at edonvarious fruits and vegetables fumigation to replace MB. Aphids are known as quarantine pest that are hard to control when conduct short period fumigation with PH3 and low dosage of EF. In this paper, dose response assessment of EF and PH3 are presented for three different aphid pecies : cotton aphid(Aphisgossypii), green peach aphid(Myzuspersicae) and turnip aphid (Lipaphiserysimi). The LCt99% values of EF at room temp. and low temp. (5℃) were 4.42 and 4.45 g·h·m-3 for cotton aphid, 3.23 and 5.58 g·h·m-3 for turnip aphid, 3.23 and 5.58 g·h·m-3 for green peach aphid when 2-hours fumigation. PH3 showed 0% efficacy on all species when 2-hours fumigation.

Methyl bromide (MB), as a fumigant for Quarantine and Pre-Shipment(QPS) could offer eradication of target pests within shorter fumigation period and without phytotoxicity. Therefore, unlike MB alternatives for soil fumigation, there is no ideally MB alternative fumigant for QPS purpose, particularly for perishable commodities. It is critically important that within shorter fumigation time requires killing all target insect pests and without effect of quality and deliver treated fruit and vegetables to the final consumer. The citrus mealybug(Planococcuscitri), problem quarantine pest, was tested with different exposure times(2 and 4-hours) and concentrations(EF(0-200 g/m3) and PH3(10-50 g/m3)) in small desiccators for mixture tests. On sensitivity test with small desiccator, the egg stage showed 100% mortality with 200/25(EF/PH3) g/m3 and the nymph and adult stage showed 100% mortality with 10/10(EF/PH3) g/m3 on 2h. Also, the egg stage showed 100% mortality with 150/25(EF/PH3) g/m3on 4h.

The Japanese termite, Reticulitermes speratus Kolbe, is distributed in Korea widely. Although, termites are not currently a great problem in Korea, it might be increased commercial losses from imported timber. Phosphine (PH3) is very effective fumigant and is widely used to control pests. Oxygen treatment was found to enhance phosphine toxicity and reduce fumigation time against various life stage of insects. In this study, we determined efficacy of oxygenated phosphine fumigation for controlling R. speratus. Fumigation to adults of R. speratus was carried in a desiccator system at 20.9% (normal), 50% and 80% of oxygen concentration for 24 h. Fumigations under higher oxygen levels greatly increased phosphine toxicity to R. speratus. Mortality of termite was increased 15.0% and 16.2% in the 50.0% and 80.0% oxygen concentration with 0.25 mg L-1 phosphine at 5℃, respectively. 100% mortality was determined in 80.0% oxygen concentration with 0.5 mg L-1 phosphine at 5℃. Oxygenated phosphine fumigations have marked potential to improve insecticidal efficacy against R. speratus. These results merit further study as potential fumigant for control of wood destroying pests.

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.

Human GH (hGH) has been available over 40 years for the treatment of children with GH deficiency. Human growth hormone (hGH) is mainly produced in the somatotrophic cells of the pituitary in brain and is the product of the GH-N gene. Among the edible mushrooms, the king oyster mushroom (Pleurotus eryngii ) is one of the most popular mushrooms in Asia, Europe and North America. The increasing popularity of P. eryngii among consumers is due to its flavor, texture and shelf life. We report a modified Agrobacteriummediated method for the efficient transformation of hGH2 in Pleurotus eryngii . The binary vector pCAMBIA1304 was used for the initial transformation and detected by GUS and GFP. Infiltrated samples transformed with pCAMBIA1304 showed a wider GUS response than the co-cultivated in the 50㎍/㎖ hygromycin selection medium. This transformation technique offers new prospects for the production of useful protein in the genetically improved mushroom.