• 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.

• The Korean Journal of Soil Zoology
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• v.5 no.2
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• pp.97-100
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• 2000

We report here an improved collecting method of the infective juveniles multiplied in the host insect with the entomopathogenic nematodes, Steinernema carpocapsae Weiser. The specific characteristics of this method involved the opening of the host insect hemocoel after the population of their infective juveniles reached at the maximum (6 days at 25$\^{C}$ after nematode treatment to nonimmunized host insects) to facilitate the escape of the multiplied nematodes. It also used 'Baermann funnel'method to select the infective juveniles effectively. This improved 'Baermann funnel'method was compared with a traditional collecting method, which was characterized with a combination of untreated host insects and 'White trap'collecting method, in both yield and pathogenicity of the collected infective juveniles to the fifth instar larvae of beet armyworm, Spodoptera exigua (H bner). More than 95% of the nematode populations collected by the two methods represented the morphological infective juveniles. To prove the nematodes to be infective juveniles functionally, pathogenicity and infective activity were compared in the nematodes collected by the two methods. They were not different in both pathogenicities and infective activities which were measured by the numbers of nematodes penetrated into the hemocoel of the insect hosts after exposure for the specific times to the same dote of infective juveniles. Significant difference between two collecting methods was found in the total yields of the infective juveniles per host insect About 50,000 infective juveniles per infected fifth instar larva of S. exigua after 6 day incubation at 25$\^{C}$ were collected only for 2 days by the improved 'Baermann funnel'method, while about 20,000 infective juveniles per host were collected for 10 days by the classical 'White trap'method.

• Korean journal of applied entomology
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• v.41 no.3
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• pp.217-223
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• 2002

The chestnut brown chafer, Adoretus tenuimaculatus Waterhouse, is serious insect pests in golf courses. Adults feed on the leaves of latifoliate trees but larvae feed on roots of turfgrases such as bentgrass, Agrostis spp. Damage of A. tenuimaculatus larvae was observed at the Jinju golf club which showed damage symptom on perennial ryegrass, Lolium perenne in tees and fairways in July, 2000. Damaged turf by A. tenuimaculatus larvae became yellowish and wilted. Symptom of laval damage of A. tenuimaculatus was similar to summer depression in warm season turfgrasses but not recovered by irrigation when Korean isolates of entomopathogenic nematodes were evaluated for the control of A. tenuimaculatus larvae in laboratory and field as a possible biological control agent. The nematodes used were Heterorhabditis bacteriophora Jeju strain, Hererorhabditis sp. Gyeongsan strain, Steinernema carpocapsae Pocheon strain, S.glaseri Dongrea strain, and S.longicaudum Nonsan strain. In the laboratory test H.bacreriophora Jeju strain and Heterorhabditis sp. Gyeongsan strain were highly effective for 3rd instars with 95% mortality. In the field test reduction rates of A.tenuimaculatus larvae were higher by ranging from 28 to 57% by H. bacteriophora Jeju strain, Heterorhabditis sp. Gyeongsan strain, and S.carpocapsae Pocheon strain compared to 7% by natural cause.

• KSBB Journal
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• v.15 no.1
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• pp.100-105
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• 2000

Optimal medium composition, culture conditions, characteristics of phase variation and activity of insecticidal toxin by Xenorhabdus nematophilus isolated and identified from Korean entomopathogenic nematode Steinernema carpocapsae were examined. Optimal medium composition of this strain was 50-70 g/L yeast extract, 3 g/L $K_{2}HPO_{4}$, 1g/L $NH_{4}H_{2}PO_{4}$, 2g/L ${MgSO}_4$$\cdot$${7H}_{2}O$, 10g/L NaCl and, these, yeast extract was found as a limiting nutrient for cell growth. When Monod equation was applied, maxmum specific growth rate and Monod constant were estimated as 0.13 $hr^{-1}$ and 20g/L, respectively. The pH of culture medium increased up to 8.5-9.5 regardless of initial pH 6-7 as the cells continued to grow. The specific growth rate in a 7 L fermentor was 0.18 $hr^{-1}$, which was enhancement 1.4 fold compared to a flask culture. In case of phase variation, phase I fraction was maintained above 90% at the stationary phase for both flask and fermentor cultures. According to oral toxicity test of Gallena mellonella by Xenorhabdus nematophilus, the addition of cell pellets into feed inhibited normal growth of insect larvae and killed completely then after 20 days cultivation. When culture supernatant of this strain was injected into hemolymph of insect larva, the toxicity was strongest at 24hr cultivation in the early exponential phase and gradually decreased as the culture time proceeded.

• Korean journal of applied entomology
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• v.35 no.1
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• pp.80-84
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• 1996

Infectivity of entomopathogenic nematodes and fungus was evaluated against fly larvae in the laboratory and outhouses. Mortalities of Muscina stabulans larvae were 96.7f 2.8% in Steinernema glaseri Dongrae strain, 90.0+0.0% in S. carpocapsae All strain, 86.7f 2.7% in Heterorhabditis bacteriophora Hamyang strain, and 70.0+9.4% in S. carpocapsae Pocheon strain on the filter paper. When 260, 000 nem\ulcornertodes were sprayed into the outhouses, H. bacterwphora Hamyang strain killed 100%, S. glaseri Dongrae strain killed 76.9+3.9%, and S. carpocapsae Pocheon strain killed 58.5+6.1% of maggots. When 130, 000 nematodes and 7.0X lo9 cfu of entomopathogenic fungus, Beauveria brongniartii were sprayed alone or combined into outhouses, mortalities of maggots were 73.6+0.1% in B. brongniartii alone, 77.8+3.9% in S. carpocapsae Pocheon strain plus B. brongniartii, and 77.7f 5.1% in H. bacteriophora Hamyang strain plus B. brongniartii. Entomopathogenic nematodes and fungus were potential biological control agents in this study.

• KSBB Journal
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• v.14 no.3
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• pp.303-308
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• 1999

An in vitro culture method for entomopathogenic nematode Steinernema glaseri was developed. A symbiotic bacterium was isolated from Steinernema glaseri and identified as Xenorhabdus nematophilus. Phase variation that differed in some biochemical characteristics of symbiotic bacterium was observed. Entomopathogenic nematodes carried only phase I bacterium in their guts. Phase I bacterium could be converted into phase II form in in vitro culture medium consisting of 5% yeast extract, 0.5% NaCl, 0.05% $K_2HPO_4$, $0.02% MgSO_4$.$7H_2O$. The optimum temperature for bacterial growth was $28^{\circ}C$. The pH of the culture medium increased up to 9.0-9.5 during the exponential growth period of the culture, regardless of initial pH 6-7. Various culture media such as chicken offal, dog food, bovine liver, peanut, and so on were tested for in vitro culture of the nematodes. The best medium for Steinernema glaseri production was obtained from concentrated homogenate of bovine liver and the nematode growth was highest at 80% bovine liver. In the co-culture of entomopathogenic nematode with its symbiont, the growth rate of nematodes was 2 times faster than that without its symbiont and the nematode concentration reached about $5.5\times10^4$/mL within 15 days.

• Weed & Turfgrass Science
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• v.4 no.1
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• pp.58-64
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• 2015

The black cutworm, Agrotis ipsilon (Hufnagel) is a major insect pest of economic crops including turfgrasses on golf courses. The entomopathogenic nematode, Steinernema carpocapsae GSN1 strain (ScG), a Korean isolate, is an effective biological control agent for soil dwelling and greenhouse insect pests in Korea. In addition, ScG is commercially produced in Korea. We conducted laboratory, greenhouse, and field trials to evaluate efficacy of ScG against black cutworms in turfgrasses. A rate of 63 infective juveniles (Ijs) per larva killed >90% of $3^{rd}$ instars feeding in cups of artificial diet within 3 days. In greenhouse trials against cutworms feeding in pots of turfgrass, efficacy of ScG was higher against $4^{th}$ instars than against $2^{nd}$ instars (90.0 vs 81.2% mortality, respectively, at $2,000Ijs\;pot^{-1}$) in perennial ryegrass, and higher against $3^{rd}$ instars in creeping bentgrass, Agrostis palustris than in zoysiagrass, Zoysia japonica (96.7 vs 52.5% mortality at $100,000Ijs\;m^{-2}$) in pot. The corrected mortality of $4^{th}$ instar was 79.9% at the rate of $100,000Ijs\;m^{-2}$ in the creeping bentgrass in the field. So ScG could be used as biological control agent against black cutworm in turfgrass of golf courses.

• Korean journal of applied entomology
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• v.42 no.2
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• pp.145-152
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• 2003

Field application of the entomopathogenic nematode, Steinernema carpncapsae, is limited by its susceptibility to UV irradiation and desiccation especially at leaf spray control. This study was conducted to develop the control technique using alginate biocapsulation of the nematodes against the beet armyworm, Spodoprera exigua and the tobacco cutworm, Sp. litura that are normally infesting hosts above ground level. The alginate capsules including infective juveniles gave significant feeding toxicities to the larvae of the two lepidopteran species. The lethality followed a typical sigmoid dose-mortality pattern with increase of the nematode densities embedded in the capsules. Moisture content in the capsule was critical to the survival of the infective juveniles. More than 80% nematodes could survive above 10% moisture content remained in the capsule. Remaining moisture content within the capsule was dependent on relative humidity, ambient temperature, and capsule size, but not on citric acid reaction time during capsule formation. More than 80% of infective juveniles in the alginate capsules could survive in distilled water at 15$^{\circ}C$ for 60 days. When these nematode capsules containing welsh onion extract as another phagostimulant were applied on the 3rd instar larvae of Sp. exigua infesting peanut plants, they resulted in about 90% control efficacy. These results indicate that the alginate capsulation can be used for leaf-spray agent of the entomopathogenic nematodes as well as for improved storage purpose.

• Korean journal of applied entomology
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• v.40 no.3
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• pp.259-264
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• 2001

Toxicological studies of two potential biological control agents, the entomopathogenic nematode (Steinernema carpocapsae) and the symbiotic bacteria (Xenorhabdus nematophilus) were conducted against two beneficial insects and one mammal species. Two microbial agents varied in their toxicities between two insect species: an ant, Pristomyrmex pungens, and silkworm, Bombyx mori. In oral toxicity test, the symbiotic bacteria resulted in significant lethal [half lethal concentration of $1.4$\times$10^3$colony-forming units (cfu)/ml] on the ants, while they gave little lethal effect (half lethal concentration of more than $10^{8}$ cfu/ml) on the silkworms. The nematodes, however, gave significant lethal effect [half lethal concentration of 4 infected juveniles (IJs)/ml] on the silkworms, while they did little lethal effect (half lethal concentration of 150,000 IJs/ml) on the ants in topical assays. Both the nematodes and the bacteria did not give lethal effect to the albino rats, Rattus norvegicus, when they were fed orally into the rats. Also, any of these microbial agents were not detected in the internal organs of the treated rats.

• Korean journal of applied entomology
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• v.27 no.4
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• pp.185-189
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• 1988

Laboratory trials were conducted for control of onion maggot and tobacco cutworm with ento¬mogenous nematodes. The onion maggot, Delia antiqua, was exposed to Steinernema feltiae at concentration of 0,30,60, 120 or 240 nematodes per larva, and to Heterorhabditis heliot-hidis at concentration of 0, 10, 20, 40 or 80 nematodes per larva. Mortalities of the maggot ranged from 80 to 100% in S. feltiae and from 63.3 to 100% in H. heliothidis. The tobacco cutworm, SpodoPtera litura was exposed to S. feltiae at concentration of 0,50,100,200 or 400 nematodes per larva and to H. heliothidis at concentration of 0,20,40,80 or 160 nemat¬odes per larva with or without kale in petri dish. The 3rd instar larvae of the tobacco cutworm was more susceptible to both nematode species than the 4th or 5th instar at low concentration. Mortalities of the 3rd instar were 100% in S. feltiae and 67.7-100% in H. heliothidis while those of 4th and 5th instar ranged 76.7-100% and 43.3-100% in S. feltiae, and 36.7-90% and 3.3-90% in H. heliothidis, respectively. Mortalities of the tobacco cutworm larvae decreased when the nematodes were sprayed on the kale leaves in petri dish except 3rd instar.