• The Korean Journal of Soil Zoology
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• v.1 no.2
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• pp.81-88
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• 1996

Entomopathogenic nematodes, Steinernema carpocapsae, was able to invade and kill the several lepidopteran pests including the beet armyworm, Spodeptera exigua Hubner, which was the most effective target host. The beet armyworms treated with the effective nematode concentrations were died within 48 hrs. The lethal effect of the nematode was varied among the developmental stages of the host. The fifth instar larvae of the beet armyworm was more vulnerable to the nematode than the third instar larvae. Pupae was, however, refractory to the nematode. All three bioessays (topical application, filter paper test, and soil treatment) showed the positive correlation between the number of the treated nematodes and the mortality of the host. Topical application was the most effective and fast-acting method so that it gave the lethal effect 2 days earlier than did filter paper test at the same number of the treated nematodes. Soil treatment required higher number of the nematodes to get the effective lethality than did filter paper test. The fifth instar larvae of the beet armyworm expressed the specific hemolymph proteins of 5 to 10 kDa in response to nematode infection.

• The Korean Journal of Ecology
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• v.22 no.5
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• pp.255-263
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• 1999

Entomopathogenic nematodes were isolated through the investigation of soil samples from various biotopes in south Korea, the efficiency of isolation for highly pathogenic nematodes to silkworms (Bombyx mori) was as high as 28 %. Twenty-eight strains of nematodes, selected among 100 samples by silkworms were confirmed the pathogenicity, multiplicity, and tolerance against various condition of preservation. Pathogenicity of the nematode isolates to agricultural and environmental pests such as Calliphora vomitoria, Pseufaletia separata, Palomena angulosa, and Melolontha incana were high. Mortality was varied from 20 to 100% by the pest insects and nematode strains. The high detectablity of entomopathogenic nematodes resulted from the methods of collection for soil samples within 10 cm depth after eliminating dried soil surface and the use of silkworm trap. High population of entomopathogenic nematodes represented the strong activity and broad action radius in the environment. Most of the nematode isolates were successfully cultured on the silkworm host as well as on artificial media, and proved their potential for the use of biological control agent.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.2
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• pp.141-145
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• 2000

Entomopathogenic nematodes are being used for insect control. We purified a toxin secreted by the insect-pathogenic bacterium, Xenorhadbus nematophilus, which lives in the gut of entomopathogenic nematodes. Culture broth of X. nematophilus was separated by centrifugation and concentrated by ultration. The concentrated culture broth was applied to a DEAE Sephadex A-50 column, and proteins were eluted stepwise with increasing concentrations of KCI. Fractions column. The molecty weight of purified toxin was39 kDa on SDS-PAGE, and Fourier tranformed infrared (FTIR) spectroscopy indicated that this toxin could be a new protein exhiting the charactristics of C=O stretching peak near 1650cm-1.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.5
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• pp.379-382
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• 2004

Entomopathogenic nematodes are used for insect control. Herein, an extracellular protease was partially purified from a culture supernatant of Xenorhabdus nematophilus, a symbiotic bacterium of an entomopathogenic nematode, Steinernema glaseri: using precipitation with $80\%$ v/v isopropyl alcohol followed by gel permeation chromatography with a packed Sephacryl S-300 HR media. The partially purified protease exhibited maximal activity at pH 7 in the presence of 1 mM $CaCl_2$. The protease was identified as a metallo-protease based on the inhibition of its activity by the metal chelating agent, EDTA.

• Journal of Sericultural and Entomological Science
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• v.40 no.2
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• pp.117-125
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• 1998

Entomopathogenic nematodes, Heterorhabditidae and Steinernematidae, were isolated from the soil of mulberry field, and the high infectivity and invesiveness were confirmed in the silkworm, Bombyx mori. The cause of non-microbial and acute flacherie was found as an disease by infection with soil-born nematodes through the mulberry leaves contaminated with soil and rainwater. The causal nematodes were isolated by silkworm trap from all of the 5 soil samples collected on the 5 mulberry fields, and identified as 3 strains of Heterorhabditis sp. and 2 of Steinernema sp. Rainwater itself, however, wasn't engaged in the silkworm disease, mulberry leaves with rainwater was rather profitable for cocoon production when the leaf quality was too hard to feed silkworm. Feeding of wet mulberry leaves with rain might not so harm to silkworm when the condition of rearing room to be kept at suitable temperature and ventilated well. Nematode infection of silkworm could be occurred by harvesting and feeding of contaminated mulberry leaves on the weather condition of rainy and wind. For the prevention of nematode infection, silkworms should be fed the leaves harvested from the higher portion of the mulberry tree in rainy days. For an oppositional application of this susceptibility of silkworms to nematode, might be useful on the collection and amplification of nematode agents for biotic control of pest insects.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.443-444
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• 2000

An in vitro liquid culture media for the cultivation of entomopathogenic nematode Steinernema carpocapsae was developed. Supplementation of whole milk powder with basal liquid culture media showed a remarkable increase in productivity compared to that without whole milk powder and the maximum nematode concentration reached about $1.5{\times}10^5/mL$ within 20 days. Five to twenty gram per liter of liver extract addition revealed highest pathogenicity against 3rd instar of Galleria mellonella which was above about 90% mortality after 48 hr.

• The Korean Journal of Soil Zoology
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• v.8 no.1_2
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• pp.17-22
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• 2003

An endemic entomopathogenic nematode, Heterorhabditis megidis, was evaluated by its control efficacies against housefly, Musca domestica, and flower beetle, Gametis jucunda. In Petri-dish assay, the pathogenicity of H. megidis showed 456.4 infective juveniles/larva (IJs/larva) in median lethality (LC$_{50}$) against the second instar larvae of M. domestica and 238.9 IJs/larva against the second instar larvae of G. jucunda. This was contrasted with those of the other well-known entomopathogenic nematode, Steinernema carpocapsae, which showed 115.9 IJs/larva against M. domestica and 388.6 IJs/larva against G. jucunda. In field experiment, H. megidis were applied per square meter of pork farm with 1,000,000 IJs of H. megidis or apple orchard with 370,000 IJs, which were infested with M. domestica or G. jucunda, respectively. H. megidis showed 56.9% and 57.3% of control efficacies against M. domestica and G. jucunda, respectively. These results suggest a promising control technique in the field using H. megidis against M. domestica and G. jucunda.a.

• Korean journal of applied entomology
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• v.39 no.3
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• pp.149-152
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• 2000

Cryopreservation of infective juveniles of entomopathogenic nematode, Steinernema carpocapsae Weiser, was conducted at $-190^{\circ}C$ liquid nitrogen and its, efficacy was analysed on nematode survival and pathogenicity with glycerol pretreatments and storage periods. Infective juveniles were pre-treated before being frozen by incubating the nematodes in 22% glycerol for each of 6, 12, and 24 h, followed by 70% methanol at $0^{\circ}C$ for 10 minutes. Just after glycerol and methanol incubations, subsamp1es of the nematodes were resuspended in 0.85% saline and maintained during 24h for viability determination. Different glycerol incubation periods significantly affected the nematode susceptibility to methanol infiltration. Six hour incubation in glycerol resulted in much less nematode survival than did 12 h or 24 h incubation. About 70% of the infective juveniles frozen at $-190^{\circ}C$ for 5 months, preincubat-ed in glycerol at least for 12h, were able to survive after being resuspended in 30°C saline. They did not also show any change in their pathogenicity during cryopreservation. These results suggest an improved technique for long-term storage of the entomopathogenic nematodes.

• Korean journal of applied entomology
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• v.41 no.3
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• pp.225-231
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• 2002

The entomopathogenic nematodes, Steinernema carpocapsae All strain (ScA), S.glaseri NC strain (SgN) and H. bacteriophora NC 1 strain (HbN), were evaluated for the effects on egg mass formation by the northern root-knot nematode, Meloidogyne hapla in pot experiment using tomato. In the first experiment, 2.5$\times$10$^{5}$ infective juveniles (Ijs) of entomopathogenic nematodes were inoculated to 100 g of the soil infected with ca. 450 Ijs of M. hapla/100 ㎤ in 150 $mell$ container. The number of egg mass was significantly decreased to 9.4-36.5 in ScA, to 5.7-24.7 in SgN and to 11.2-16.0 in HbN treatments compared with 62.5 in M.hapla alone. In the second experiment, ScA and S.carpocapsae Pocheon strain (ScP) and SgN and S.glaseri Dongrae strain (SgD) were treated to 350 g of the soil infected with 100, 200 M.hapla larvae/100 ㎤ in 450 $mell$ container The entomopathogenic nematodes were inoculated at the rate of 2,020 Ijs and 1.6$\times$105 Ijs in 350 g soil. The number of egg mass of M.hapla were significantly decreased in the entomopathogenic nematode treatments compared with M.hapla alone although no differences were observed among Steinernema species, strains, or infection concentrations. Treatments of entomopathogenic nematodes 3 days before M.hapla inoculation were more effective on reduction of egg mass formation than those of 3 days after M.hapla treatments. Growth of tomato was not affected by entomopathogenic nematode treatments.

• KSBB Journal
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• v.14 no.2
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• pp.198-204
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• 1999

Asymbiotic bacterium with highly effective toxins was isolated from entomopathogenic nematode Steinernema glaseri which has been widely used against various soil-inhabiting pests. The symbiont of S. glaseri was identified as Xenorhabdus nematophilus sp. by using several biochemical and physiological tests. When this strain was released into the hemolymph of insect larva, it produced highly toxic substances and killed the larva within 2 days. Two colony forms that differed n some biochemical characteristics were observed when cultures in vitro. Phase l colonies were mucid and difficult to be dispersed in liquid. Phase II was not mucoid and was easily dispersed in liquid. It did not adsorb neutral red or bromothymol blue. Rod-shaped cell size was highly variable between two phases, ranging 2-10 ${\mu}{\textrm}{m}$. It was also found that only infective-stage nematodes can carry only primary-phase Xenorhabdus in their intestine.