• Korean Journal of Environmental Biology
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• v.17 no.3
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• pp.321-330
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• 1999

Nematodes were isolated using silkwom trap through the investigation of 100 soil samples from various biotopes in Korea. The 30 nematode strains from soil and dead insects by the pathogenicity aganinst silkworms (Bombyx mori mori) and insect pests of Calliphora vomitoria, Pseufazetia separata, Palomena angulosa, and Melolontha incana. Mortailty of the silkworm larvae and pupae were as high as 100% by nematode infection, those of insect of pests were varied from 20 to 100%. The 30 strains of entemopathogenic nematodes were classified into five groups of Rhabditidae, Diplogatroidae, Heterorhabitidae, Steinernematidae, and Tylenchida by morphological criteria. The genetic relationships among the 30 nematode strains were analyzed by various RAPD bands with twenty primers. The 30 nematode strains were classified into six major subgroups on the basis of the genetic similarity coefficient of 0.853. The grouping by RAPD was agree with those of morphological taxa in discrimination of the higher group, however, was not completely agree in the subgroup. The family Steinernematidae belong to Rhabditida was clarified as closer to the Tylenchida, rather than the other Rhabditida of Heterorhabitidae, Rhabditidae, and Diplogatroidae in genetic distance valule. From the result of the morphological classification and RAPD of the genomic DNA showed that genetic relationship analysis furnish infurmation on phylogenetic classification and relationships of entomopathogenic nematodes. The application of genetic similarity will overcome the limitation of taxonomy and classification of morphologically simple nematode. Several primers were confirmed those utility of identification for individual nematode strains, the methods of molecular genetics secured the simplicity, rapidity and accuracy on the selection of entomopathogenic nematodes.

• Korean journal of applied entomology
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• v.49 no.1
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• pp.61-67
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• 2010

Effect of inoculation level on pathogenicity, development, and propagation of entomopathogenic nematode, Steinernema arenarium was investigated using the last instar of great wax moth, Galleria mellonella. Pathogenicity of S. arenarium was higher with increasing inoculation level representing 82% at the rate of 5 infective juveniles (IJs) while >98% at the rate of >10 IJs. The number of IJs penetrated into the host was 2.7, 5.0, 7.4, and 12.2 at the rate of 5, 10, 20, and 40 IJs, respectively while 24.3 at the rate of 80 IJs and 40.2 at the rate of 160 IJs. Inoculation level did not affect female adult size (4,616 to 6,444 ${\mu}m$) while affected male adult size (1,600 to 1,934 ${\mu}m$). The rate of stunted female adults was 70.2% at the inoculation level of 80 IJs and 63.7% at the inoculation level of 160 IJs. The number of progenies was 20,431, 26,696, 47,943, 50,516, 58,701, and 74,235 at the rate of 5, 10, 20, 40, 80, and 160 IJs, respectively. The body lengths of IJs were different depending on inoculation level ranging from 636 to 1,496 ${\mu}m$.

• Journal of agriculture & life science
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• v.45 no.6
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• pp.115-124
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• 2011

This study was conducted to find a useful alternative herbivore system with which to study the effects of ultraviolet exposure on the pathogenicity and survival of Korean isolated entomopathogenic nematodes (Heterorhabditis sp. 202 strain, Heterorhabditis sp. Gyeongsan strain, Steinernema sp. 223 strain, S. carpocapsae Pocheon strain, S. glaseri Dongrae strain and S. longicaudum Nonsan strain). Pathogenicity of entomopathogenic nematodes against silkworm, Bombyx mori, differed depending on the nematode species and strain and instar of silkworm challenged. Steinernematidae were relatively more pathogenic to the 5th instar silkworms, whereas Heterorhabditidae were more pathogenic to the 1st instars. UV-C exposure was harmful to entomopathogenic nematodes. All nematodes were killed within 60 minutes of exposure of UV-C, and after 10 min of exposure they caused only 6.7% corrected mortality of silkworms. Silkworms were not infected when they were fed S. carpocapsae Pocheon strain treated mulberry leaves which were exposed to UV intensity of about $2.3mW/cm^2$ for 4 hours in the field. Pathogenicity of S. carpocapsae was not significantly reduced after 1 hour of exposure to $4.0mW/cm^2$ UV intensity on the mulberry leaves against silkworms.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.906-909
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• 2001

In order to investigate fatty acid contents and effects of cell growth on the production of an extracellular protease and toxicity of exotoxin, several symbiotic bacteria with highly effective toxins were isolated from seven species of entomopathogenic nematodes belong in Steinernematidae(Steinernema glaseri XR-DR, S. glaseri XR-NC, S. glaseri XR-MK, S. carpocapsae XR-PC, S. maticola XR-MO, S. Longicaudum XR-LC) and Heterorhabditidae sp.(Heterorhabditis bacteriophora XR-HY). In the cell growth and exotoxin toxicity, XR-PC and XR-MK were superior to other species when cultured in vitro. The protease activity of XR-DR was remarkable compared to other species. In the case of XR-HY, the protease activity increased in parallel with cell growth. Interestingly the fatty acid contents of XR-PC and XR-HY were significantly different from those of other species 12:0, 14:0, 13:0 iso, 16:1 cis 5 and 17:0 cyclo.

• 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.

• The Korean Journal of Soil Zoology
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• v.1 no.1
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• pp.28-36
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• 1996

A survey for entomopathogenic nematodes was conducted throughout the nine provinvrd and within three city limits during the summer of 1990 and 1991. Six of the nine provinces and one of the three cities were positive for entomopathogenic nematodes. Out of the total 499 soil samples, 23(4.6%) were positive for entomopathogenic nematodes with 19(3.8%) containing Steinernema and 4(0.8%) containing Heterorhabditis. Heterorhabditis bacteriopora and three distinct groups of Steinernema species were idintified. One groups was indentified as S. carpocapsae, another S. glaseri and the other S. monticola based on cross breeding studies. Positive sample sites in each habitat includes 15 of the 415(3.6%) from forests including regrowth areas with shrubs, 1 of the 27(3.7%) from turfgrass including golf courses and parks, 3 of the 24(12.5%) from agricultural fields, 2 of the 16(12.5%) along riparian areas, and of the 17(11.8%) near the seashore. We advocate that more surveys be conducted for entomopathogenic nematodes before commercial sources of nematodes are widely applied which may obscure the naturally-occurring nematodes. A key to Steinernema is provided for the identification.

• KSBB Journal
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• v.17 no.3
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• pp.276-282
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• 2002

Symbiotic bacteria with highly effective insecticidal activities were isolated and compared with their physiological characteristics from seven species of entomopathogenic nematodes belong to Steinernamatidae and Heterorhabditidae sp., and three of them were identified as Xenorhabdus nematophilus. Culture characteristics, insecticidal activities, pretense activities and fatty acid contents of various symbiotic bactierial isolates were also examined. In the case of cell growth and insecticidal activity, XR-PC and XR-MK were superior to other species when cultured in vitro. The insecticidal activity were highest at the early exponential growth phase, and gradually decreased with time. The protease activity of XR-DR was remarkable compared to other species. In the case of HE-HY, however the pretense activity increased in parallel with cell growth. Interestingly, the fatty acid patterns of Xenorhabdus nematophilus isolated from different emtomopathogenic nematode, showed remarkable differences in their contents of 12:0, 14:0, 16:1 cia 5 and 17:0 cyclo and hydroxy and branch factty acids were varied from 2% to 15% among total fatty acid contents.

• 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.

• Korean journal of applied entomology
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• v.30 no.1
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• pp.50-53
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• 1991

The entomopathogenic nematodes, Steinernema carpocapsae and Heterorhabditis bacteriophoTa, were laboratory tested for the control of rice stem borer Chilo suppressalis. When 8 cm long of two rice stems infected by rice stem borers were treated with S. carpocapsae by spray application to the concentrations of 250, 500, 1,000, or 2,000 nematodes/ml, 68.6 $\pm$ 10.0-94.9 $\pm$ 2.6 % control was obtained. When 8 cm long of rice stems infected by rice stem borers were treated with H. bacteriophoTa by spray application to the concentrations of 100, 200, 400, or 800 nematodes/ ml, 91.4 $\pm$ 0.7-100 % control was obtained. On the other hand, when 8 cm long of three rice stems were treated with H. bacteriophora by dipping application to the concentrations of 100 or 200 nematodes/ml, 46.2 $\pm$ 4.7-63.1 $\pm$ 4.7 % control was obtained. Because the moist habitat of rice stems were favorable to nematode survival and searching abilities, entomopathogenic nematode, were confirmed to be a potential biological control agents against rice stem borers.

• Korean journal of applied entomology
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• v.41 no.4
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• pp.269-277
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• 2002

Ecological studies on entomopathogenic nematodes are required to increase control efficacy against target insect pests and to obtain basic information for mass production. Thus, effect of temperature and nematode concentration on infectivity and reproduction of Steinernema carpocapsae Pocheon and that of exposure time and soil depth on infectivity were examined using Galleria mellonella larvae. Infectivity and reproduction were examined at five temperatures, 13, 18, 24, 30 and 35$^{\circ}C$ with seven concentrations, 0, 5, 10, 20, 40, 80 and 160 infective juveniles (IJs)/larva. Temperature and nematode concentration influenced infectivity and reproduction of S. carpocapsae Pocheon. Although G. mellonella larvae were killed by S. carpocapsae Pocheon at all given temperatures and nematode concentrations, mortality was higher at 24$^{\circ}C$ than other temperatures. Lethal time of G. mellonella by S. carpocapsae Pocheon was shorter with increasing temperature and nematode concentrations. S. carpocapsae Pocheon was not established in G. mellonella at 13 and $35^{\circ}C$. Time for the first emergence from G. mellonella cadaver was longer $18^{\circ}C$ (about 20 days) than 24 and $30^{\circ}C$ (about 5 days). The highest number of progenies was obtained at $24^{\circ}C$ with 80IJs/1arva, i.e., $18.8$\times$10^4$IJs were produced from a larva. In the exposure time assay, G. mellonella death was recorded in 10 minutes when 300 IJs were inoculated per larva. When S. carpocapsae Pocheon was inoculated at the rate of $10^{9}$ IJs/ha to G. mellonella at the depth of 0, 2, 5 and 10 cm of sand columns, 100% mortality and similar sex ratio were observed but number of established IJs in cadaver was decreased with deepening the soil depth. The results indicated that optimum temperature for infectivity and reproduction of S. carpocapsae Pocheon was $24^{\circ}C$ In addition, S. carpocapsae Pocheon was effective to target insects within 5 cm from the soil surface.