• Journal of Microbiology and Biotechnology
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• v.17 no.3
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• pp.420-427
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• 2007

We investigated the temperature effects on the virulence, development, reproduction, and otility of two Korean isolates of entomopathogenic nematodes, Steinernema glaseri Dongrae strain and S. longicaudum Nonsan strain. In addition, we studied the growth and virulence of their respective symbiotic bacterium, Xenorhabdus poinarii for S. glaseri and Xenorhabdus sp. for S. longicaudum, in an insect host at different temperatures. Insects infected with the nematode-bacterium complex or the symbiotic bacterium was placed at $13^{\circ}C,\;18^{\circ}C,\;24^{\circ}C,\;30^{\circ}C,\;or\;35^{\circ}C$ in the dark and the various parameters were monitored. Both nematode species caused mortality at all temperatures tested, with higher mortalities occurring at temperatures between $24^{\circ}C\;and\;30^{\circ}C$. However, S. longicaudum was better adapted to cold temperatures and caused higher mortality at $18^{\circ}C$ than S. glaseri. Both nematode species developed to adult at all temperatures, but no progeny production occurred at $13^{\circ}C\;or\;35^{\circ}C$. For S. glaseri, nematode progeny production was best at inocula levels above 20 infective juveniles/host at $24^{\circ}C\;and\;30^{\circ}C$, but for S. longicaudum, progeny production was generally better at $24^{\circ}C$. Steinernema glaseri showed the greatest motility at $30^{\circ}C$, whereas S. longicaudum showed good motility at $24^{\circ}C\;and\;30^{\circ}C$. Both bacterial species grew at all tested temperatures, but Xenorhabdus sp. was more virulent at low temperatures $(13^{\circ}C\;and\;18^{\circ}C)$ than X. poinarii.

• Journal of Microbiology and Biotechnology
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• v.19 no.3
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• pp.314-322
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• 2009

Four different bacterial colonies were isolated from an old stock of an entomopathogenic nematode, Steinernema monticolum. They all showed entomopathogenicity to final instar larvae of beet armyworm, Spodoptera exigua, by hemocoelic injection. However, they varied in colony form, susceptibility to antibiotics, and postmortem change of the infected host insects. Biolog microbial identification and 16S rDNA sequence analyses indicate that these are four different species classified into different bacterial genera. Owing to high entomopathogenicity and a cadaver color of infected insect host, Serratia sp. was selected as a main symbiotic bacterial species and analyzed for its pathogenicity. Although no virulence of Serratia sp. was detected at oral administration, the bacteria gave significant synergistic pathogenicity to fifth instar S. exigua when it was treated along with a spore-forming entomopathogenic bacterium, Bacillus thuringiensis. The synergistic effect was explained by an immunosuppressive effect of Serratia sp. by its high cytotoxic effect on hemocytes of S. exigua, because Serratia sp. caused septicemia of S. exigua when the bacterial cells were injected into S. exigua hemocoel. The cytotoxic factor(s) was present in the culture medium because the sterilized culture broth possessed high potency in the cytotoxicity, which was specific to granular cells and plasmatocytes, two main immune-associated hemocytes in insects.

• 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.

• Korean journal of applied entomology
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• v.51 no.1
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• pp.39-45
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• 2012

The entomopathogenic bacterium, $Xenorhabdus$ sp., was isolated from an entomopathogenic nematode, $Steinernema$ $monticolum$. When these bacteria were injected into the hemocoel of the diamondback moth, $Plutella$ $xylostella$, they caused significant mortality. However, the bacterium was not pathogenic when it was administered orally. This study showed that $Xenorhabdus$ sp. significantly enhanced oral pathogenicity of $Bacillus$ $thuringiensis$ (Bt) against the last instar larvae of $P.$ $xylostella$. Different ratios of culture broth of $Xenorhabdus$ sp. and Bt showed significantly different pathogenicities against $P.$ $xylostella$. In field tests, the optimal bacterial mixture significantly enhanced control efficacy against $P.$ $xylostella$ compared to Bt treatment alone. These results demonstrated that $Xenorhabdus$ sp. culture broth can be developed as a potent biopesticide by enhancing the insecticidal efficacy of Bt.

• Journal of Sericultural and Entomological Science
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• v.50 no.2
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• pp.140-144
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• 2012

This study was aimed for identification of a useful genetic resources from the entomopathogenic bacteria infected-midgut of the silkworm, Bombyx mori L. We analyzed the appropriately midgut-immunizing condition of $4^{th}$ instar larvae by a feeding infection using several entomopathogenic bacteria. Xenorhabdus nematophila was selected as a suitable bacteria for midgut immunization of Jam 123, B. mori. We constructed a subtraction cDNA library from the mRNA of the immunized midgut, respectively. A total of 1,000 clones were randomly selected from the subtracted cDNA library, and then performed a differential display hybridization analysis with forward and reverse probes. In conclusion, nine clones were identified as differential expressed genes, which presumed that these genes were involved in gut immunity of silkworm. The total number of clones analyzed in this work is not enough to have a brief overview of a understanding on the midgut immunity factors of silkworm. Therefore, further defined studies on these molecules biological roles will give us well-fined information about the innate immune mechanism of silkworm.

• Journal of Microbiology and Biotechnology
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• v.17 no.6
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• pp.968-978
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• 2007

A pathogenic nematode, Butlerius sp., was isolated from Oriental beetle, Blitopertha orientalis. The infective juveniles exhibited dose-as well as time-dependent entomopathogenicity on the larvae of B. orientalis. Two bacterial species, Providencia vermicola (KACC 91278) and Flavobacterium sp. (KACC 91279), were isolated from the infective juveniles and identified. P. vermicola outnumbered Flavobacterium sp. in the nematode host, in which the colony density of P. vermicola was found to be 21 times higher than that of Flavobacterium sp. However, when the two bacterial species were cocultured in culture media without the nematode host, they showed similar growth rates. Both bacteria induced significant entomopathogenicity against Spodoptera exigua larvae infesting economically important vegetable crops, where P. vermicola was more potent than Flavobacterium sp.

• Korean Journal of Organic Agriculture
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• v.30 no.1
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• pp.103-118
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• 2022

Photorhabdus is a bacterial symbiont of entomopathogenic nematodes of the genus Heterorhabditis in the family Heterorhabditidae. Photorhabdus is known to have nematicidal activity in addition to insecticidal activity. P. temperata isolated from Korean indigenous H. megidis Gwangju strain also produced high control efficacy against root-knot nematode Meloidogyne incognita and root-lesion nematode Pratylenchus penetrans. P. temperata has drawn interest as a potential bionematicide for the control of root-knot nematodes thereby. For the registration as an organic agricultural material, the toxicity of P. temperata was assessed by the acute toxicity test in carp (Cyprinus carpio) and acute oral and dermal toxicity tests in Sprague-Dawley rat (Rattus norvegicus) in compliance with the guidelines of the Rural Development Administration (RDA). In the acute toxicity test in fish, neither lethality nor abnormal responses of carp were observed. Body length and weight of carp and changes in DO concentrations and pH values were not significantly different between the treated group and the untreated control. In the acute oral and dermal toxicity tests, clinical signs, abnormal behavior, mortality, and pathological findings were not observed in all the experimental rats. The weight increment of all rats was normal. Acute toxicity results of P. temperata in fish and rats belonged to categories III, IV, and IV of RDA, respectively. Toxicity results of the present study indicated that P. temperata could be a safe and promising bionematicide against root-knot nematodes and root lesion nematode.

• Korean journal of applied entomology
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• v.48 no.3
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• pp.377-383
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• 2009

This study was conducted to investigate the total phenol contents, antioxidative activities and antibacterial activities of twenty species of entomopathogenic fungi. The total phenol content was highest in Aspergillus flavus ($553.0{\pm}52.15{\mu}g/g$) and A. parasiticus ($529.9{\pm}60.10{\mu}g/g$). On the other hand those in other strains were within the range of $26.6{\sim}121.9{\mu}g/g$. The antioxdative activity was shown in the most of strains and the highest DPPH (1, 1-diphenyl-2-picrylhydrazyl) radical scavenging activity was observed in A. flavus ($90.9{\pm}2.90%$) and A. parasiticus ($77.9{\pm}4.13%$). This result indicated that the antioxidative activities were very correlated with the total phenol contents. The antibacterial activitiy was found in the every tested pathogenic bacteria. Especially, the antibacterial activity was strongest against Listeria monocytogenes and Escherchia coli.

• Korean journal of applied entomology
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• v.41 no.4
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• pp.279-284
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• 2002

Flacherie symptom was found in the fifth instar larvae of silkworm, Bombyx mori. The bacterial pathogen was isolated from the hemolymph of the infected silkworm and identified. The isolated bacteria caused a significant flacherie pathogenicity to the fifth instar larvae of B. mori when $5{\times}10^{6}$ cfu (colony-forming unit) of the bacteria was injected into each larva. The infected larvae began to die at 6 days after injection and resulted in complete mortality at 10 days. The bacterium was identified as Staphylococcus gallinarum based on the morphological and physiological characteristics described in Bergey's manual. This identification was further supported by the characters of carbohydrate utility analyzed from a bacterial identification system ($MicroLog^{\circledR}$) and also by the molecular structure of 165-23S rDNA internal transcribed spaces. As an insecticidal action, S. gallinarum caused hemolymph septicemia by its cytotoxic effect on the hemocytes of B. mori.

• The Korean Journal of Mycology
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• v.30 no.1
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• pp.44-49
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• 2002

The study was conducted to isolate and identify insecticidal bacteria for biological control of larvae of mushroom fly, Lycoriella mali, which is one of serious pests to oyster mushrooms during its cultivation period. Among eight bacteria isolated from the soil in the oyster mushroom beds and the dead body of L. mali, two bacteria, Bti-D and Bti-U showed more toxicity with mortality rate than other six-bacteria isolates. The two bacteria showed more toxicity in three instar of the period of development of the mushroom fly than in other instar. Symptoms of the larvae of L. mali infected by the two bacteria developed as follows: at the early infection, the front middle gut changed color to light brown, the middle gut to brown, whole body to black brown, and eventually, the fly died. For the identification of these isolates, cultural and biochemical characteristics by Bergey's manual and Biolog system, cell morphology by TEM, endospore and endotoxin by phase-contrast microscope, and test using 33H antisera were examined. According to the results, these two isolates, Bti-D and Bti-U were identified as Bacillus thuringiensis subsp. israelensis respectively.