• Journal of Microbiology and Biotechnology
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• v.18 no.7
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• pp.1235-1244
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• 2008

Indole-3-acetic acid (IAA) is produced commonly by plants and many bacteria, however, little is known about the genetic basis involving the key enzymes of IAA biosynthetic pathways from Bacillus spp. IAA intermediates from the Gram-positive spore-forming bacterium Paenibacillus polymyxa E681 were investigated, which showed the existence of only an indole-3-pyruvic acid (IPA) pathway for IAA biosynthesis from the bacterium. Four open reading frames (ORFs) encoding indole-3-pyruvate decarboxylase-like proteins and putative indole-3-pyruvate decarboxylase (IPDC), a key enzyme in the IPA synthetic pathway, were found on the genome sequence database of P. polymyxa and cloned in Escherichia coli DH5$\alpha$. One of the ORFs, PP2_01257, was assigned as probable indole-3-pyruvate decarboxylase. The ORF consisted of 1,743 nucleotides encoding 581 amino acids with a deduced molecular mass of 63,380 Da. Alignment studies of the deduced amino acid sequence of the ORF with known IPDC sequences revealed conservation of several amino acids in PP2_01257, essential for substrate and cofactor binding. Recombinant protein, gene product of the ORF PP2_01257 from P. polymyxa E681, was expressed in E. coli BL21 (DE3) as a glutathione S-transferase (GST)-fusion protein and purified to homogeneity using affinity chromatography. The molecular mass of the purified enzyme showed about 63 kDa, corresponding closely to the expected molecular mass of IPDC. The indole-3-pyruvate decarboxylase activity of the recombinant protein, detected by HPLC, using IPA substrate in the enzyme reaction confirmed the identity and functionality of the enzyme IPDC from the E681 strain.

• Journal of Life Science
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• v.23 no.2
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• pp.299-305
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• 2013

Paenibacillus is a gram-positive, spore-forming aerobes that was previously classified as a Bacillus species. Paenibacillus sp. CK214 was highly motile on LB agar plates and showed typical colonial morphology of Paenibacillus. However, its motility was defective in the absence of glucose. Electron microscopic observation revealed that the cells of CK214 cultured on LB agar plates were peritrichously flagellated but not flagellated in the presence of glucose. Flagellar filaments were purified by centrifugation after shearing off from the CK214 cells with vigorous pipetting. The purified protein was composed of a single flagellin with an apparent molecular size of 29 kDa. Recognition of the protein by anti-Edwardsiella tarda flagellin protein antibody demonstrates that the protein is a flagellin protein. A decreased level of flagellin protein was detected in CK214 cells grown under glucose-supplemented media.

• Korean journal of applied entomology
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• v.52 no.2
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• pp.115-123
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• 2013

A microbial insecticide "Bt-Plus" has been developed to enhance an insecticidal efficacy of an entomopathogenic bacterium, Bacillus thuringiensis (Bt). However, its wettable powder formulation is not preferred by farmers and industry producers due to relatively high cost. This study aimed to develop a soluble concentrate formulation of Bt-Plus. To this end, an optimal mixture ratio of two bacterial culture broths was determined to be 5:4 (v/v) of Bt and Xenorhabdus nematophila (Xn) along with 10% ethanol preservative. In addition, Bt broth was concentrated by 10 times to apply the mixture at 1,000 times fold dilution. The resulting liquid formulation was sprayed on cabbage crop field infested by late instar larvae of the diamondback moth, Plutella xylostella. The field assay showed about 77% control efficacy at 7 days after treatment, which was comparable to those of current commercial biopesticides targeting P. xylostella. For storage test in both low and room temperatures, the liquid formation showed a relatively stable control efficacy at least for a month. To develop a quality control technique to exhibit a stable control efficacy of Bt-Plus, Bt spore density ($5{\times}10^{11}$ spores/mL) and eight active component concentrations of Xn bacterial metabolites in the formulation products have been proposed in this study.

• Journal of Microbiology and Biotechnology
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• v.15 no.6
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• pp.1286-1298
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• 2005

The genus Paenibacillus is a new group of bacilli separated from the genus Bacillus, and most of species have been isolated from soil. In the present study, we collected 450 spore-forming bacilli from the roots of winter crops, such as barley, wheat, onion, green onion, and Chinese cabbage, which were cultivated in the southern part of Korea. Among these 450 isolates, 104 Paenibacillus-like isolates were selected, based on their colony shape, odor, color, and endospore morphology, and 41 isolates were then finally identified as Paenibacillus spp. by 16S rDNA sequencing. Among the 41 Paenibacillus isolates, 23 were classified as P. polymyxa, a type species of the genus Paenibacillus, based on comparison of the 16S rDNA sequences with those of 32 type strains of the genus Paenibacillus from the GenBank database. Thirty-five isolates among the 41 Paenibacillus isolates exhibited antagonistic activity towards plant fungal and bacterial pathogens, whereas 24 isolates had a significant growth-enhancing effect on cucumber seedlings, when applied to the seeds. An assessment of the root-colonization capacity under gnotobiotic conditions revealed that all 41 isolates were able to colonize cucumber roots without any significant difference. Twenty-one of the Paenibacillus isolates were shown to contain the nifH gene, which is an indicator of $N_{2}$ fixation. However, the other 20 isolates, including the reference strain E681, did not incorporate the nifH gene. To investigate the diversity of the isolates, a BOX-PCR was performed, and the resulting electrophoresis patterns allowed the 41 Paenibacillus isolates to be divided into three groups (Groups A, B, and C). One group included Paenibacillus strains isolated mainly from barley or wheat, whereas the other two groups contained strains isolated from diverse plant samples. Accordingly, the present results showed that the Paenibacillus isolates collected from the rhizosphere of winter crops were diverse in their biological and genetic characteristics, and they are good candidates for further application studies.

• Korean Journal of Soil Science and Fertilizer
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• v.29 no.3
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• pp.303-311
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• 1996

The microflora changes of 10 water-controled treatments combined with livestock manures(pig, chicken) and bulking agents(sawdust, paper sludge) were investigated. The B/F values of the P-1 and C-1(65%, $H_2O$) treatments were 3571 and 5400 respectively, but those of the P-4 and C-4(50%, $H_2O$) treatments showed very low values, 667 and 334, respectively. The B/F values tended to increase with higher water content of the treatments. In the composting processes, the successions of microflora, adapting the compost environments, took place via fluctuating temperature. In the high temperature period, the numbers of mesophilic bacteria and fungi decreased, but that of the spore forming bacteria increased. However, the number of mesophilic bacteria inereased during the cold period. The B/F values of compost ranged 25-300, which indicates a decrease in the quantity of bacteria. The time required for the temperature of compost to reach $60^{\circ}C$ showed different patterns. There was no pathogenic microorganism in the treatments which reached a high temperature in a short period of time, but, in the treatments which reached a high temperature over a Long period of time, the pathgenic microorganism was not still alive.

• Journal of Animal Environmental Science
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• v.8 no.3
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• pp.183-190
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• 2002

A study was conducted to determine the effect of dietary probiotics or antibiotics on growth and pathological status in growing-finishing pigs. Ninety male pigs weaned at 24 days of age were divided into three groups of 30 pigs each on the basis of body weight and litter. Three groups of ten pigs(one pen) each were assigned to one of the following diets; a control diet or diets containing 0.1% probiotics or 0.1% antibiotics (1:1 mixture of kitasamycin and sulfamethazine). Average daily gain (ADG), feed efficiency(G/F) and the pathological status were monitored. ADG, feed efficiency and carcass quality were not different (P>0.05) among the three treatments. But pork quality in pigs fed probiotics tended to be improved, compared to other treatments. The pigs fed probiotics had lower pathological lesion in intestinal monitoring than that of other treatments pigs. The chemical composition of slurry(BOD, COD, SS, T-N, T-P and ammonia) in the probiotics treatments tended to be decreased, compared to other treatments. Results of this study suggest that dietary probiotics improve pigs' housing environment, and decrease the contents of polluting materials in slurry.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.12
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• pp.1657-1667
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• 2010

Direct-fed microbials (DFM) are dietary supplements that inhibit gastrointestinal infection and provide optimally regulated microbial environments in the digestive tract. As the use of antibiotics in ruminant feeds has been banned, DFM have been emphasized as antimicrobial replacements. Microorganisms that are used in DFM for ruminants may be classified as lactic acid producing bacteria (LAB), lactic acid utilizing bacteria (LUB), or other microorganisms including species of Lactobacillus, Bifidobacterium, Enterococcus, Streptococcus, Bacillus and Propionibacterium, strains of Megasphaera elsdenii and Prevotella bryantii and yeast products containing Saccharomyces and Aspergillus. LAB may have beneficial effects in the intestinal tract and rumen. Both LAB and LUB potentially moderate rumen conditions and improve feed efficiency. Yeast DFM may reduce harmful oxygen, prevent excess lactate production, increase feed digestibility, and improve fermentation in the rumen. DFM may also compete with and inhibit the growth of pathogens, stimulate immune function, and modulate microbial balance in the gastrointestinal tract. LAB may regulate the incidence of diarrhea, and improve weight gain and feed efficiency. LUB improved weight gain in calves. DFM has been reported to improve dry matter intake, milk yield, fat corrected milk yield and milk fat content in mature animals. However, contradictory reports about the effects of DFM, dosages, feeding times and frequencies, strains of DFM, and effects on different animal conditions are available. Cultivation and preparation of ready-to-use strict anaerobes as DFM may be cost-prohibitive, and dosing methods, such as drenching, that are required for anaerobic DFM are unlikely to be acceptable as general on-farm practice. Aero-tolerant rumen microorganisms are limited to only few species, although the potential isolation and utilization of aero-tolerant ruminal strains as DFM has been reported. Spore forming bacteria are characterized by convenience of preparation and effectiveness of DFM delivery to target organs and therefore have been proposed as DFM strains. Recent studies have supported the positive effects of DFM on ruminant performance.

• Food Science of Animal Resources
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• v.28 no.1
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• pp.76-81
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• 2008

The objective of this study is to evaluate the microbial safety of raw pork used to produce Korean pork jerky. The raw pork samples harbored large populations of microorganisms. In particular, mesophilic bacteria were found to be most numerous $(3.9{\times}10^2-3.9{\times}10^5cfu/g)$ in the samples. Spore-forming bacteria and coliforms were not detected below detection limit. Yeast and molds were detected at $3.8{\times}10^1-5.1{\times}10^2cfu/g$ in the raw pork. Ten samples of raw pork were analyzed for the presence of pathogenic bacteria. Bacillus cereus was isolated from samples B and J and Staphylococcus aureus was isolated from sample B. The B. cereus isolates from raw pork samples were identified with 99.8% agreement and S. aureus isolate was identified with 97.8% agreement according to the API CHB 50 kit.

• 한국균학회소식:학술대회논문집
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• 2016.05a
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• pp.25-25
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• 2016

Biological control has many advantages as a disease control method, particularly when compared with pesticides. One of the most important benefits is that biological control is an environmental friendly method and does not introduce pollutants into the environment. Another great advantage of this method is its selectivity. Selectivity is the important factor regarding the balance of agricultural ecosystems because a great damage to non target species can lead to the restriction of natural enemies' populations. The objective of this research was to evaluate the effects of several different bacterial isolates on the efficacy of biological control of soil borne diseases. White rot caused by Sclerotium cepivorum was reported to be severe disease of garlic and chive. The antifungal bacteria Burkholderia pyrrocinia CAB08106-4 was tested in field bioassays for its ability to suppress white rot disease. In field tests, B. pyrrocinia CAB08106-4 isolates suppressed white rot in garlic and chive, with the average control efficacies of 69.6% and 58.9%, respectively. In addition, when a culture filtrate of B. pyrrocinia CAB08106-4 was sprayed onto wounded garlic bulbs after inoculation with a Penicillium hirstum spore suspension in a cold storage room ($-2^{\circ}C$), blue mold disease on garlic bulbs was suppressed, with a control efficacy of 79.2%. These results suggested that B. pyrrocinia CAB08106-4 isolates could be used as effective biological control agents against both soil-borne and post-harvest diseases of Liliaceae. Chinese cabbage clubroot caused by Plasmodiophora brassicae was found to be highly virulent in Chinese cabbage, turnips, and cabbage. In this study, the endophytic bacterium Flavobacterium hercynium EPB-C313, which was isolated from Chinese cabbage tissues, was investigated for its antimicrobial activity by inactivating resting spores and its control effects on clubroot disease using bioassays. The bacterial cells, culture solutions, and culture filtrates of F. hercynium EPB-C313 inactivated the resting spores of P. brassicae, with the control efficacies of 90.4%, 36.8%, and 26.0%, respectively. Complex treatments greatly enhanced the control efficacy by 63.7% in a field of 50% diseased plants by incorporating pellets containing organic matter and F. hercynium EPB-C313 in soil, drenching seedlings with a culture solution of F. hercynium EPB-C313, and drenching soil for 10 days after planting. Soft rot caused by Pectobacterium carotovorum subsp. carotovorum was reported to be severe disease to Chinese cabbage in spring seasons. The antifungal bacterium, Bacillus sp. CAB12243-2 suppresses the soft rot disease on Chinese cabbage with 73.0% control efficacy in greenhouse assay. This isolate will increase the utilization of rhizobacteria species as biocontrol agents against soft rot disease of vegetable crops. Sclerotinia rot caused by Sclerotinia sclerotiorum has been reported on lettuce during winter. An antifungal isolate of Pseudomonas corrugata CAB07024-3 was tested in field bioassays for its ability to suppress scleritinia rot. This antagonistic microorganism showed four-year average effects of 63.1% of the control in the same field. Furthermore, P. corrugata CAB07024-3 has a wide antifungal spectrum against plant pathogens, including Sclerotinia sclerotiorum, Sclerotium cepivorum, Botrytis cinerea, Colletotrichum gloeosporioides, Phytophotra capsici, and Pythium myriotylum.

• Korean Journal of Soil Science and Fertilizer
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• v.18 no.2
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• pp.221-226
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• 1985

This investigation was undertaken to clarify the effects of consecutive application of insecticide (Hexachlorocyclohexane: HCH, 10 ppm each year) and fungicide (Tetrachloroisophthalonitrile: TPN, 40 ppm each year) on changes of the composition of soil bacterial flora in the experimental plots treated with each pesticide for two years. For these purposes, the isolating of bacterial cells growing on albumin agar plate was carried out with non-treated, HCH-treated and TPN-treated soil. And these isolated strains were grouping in accordance with the first diagnostic table of Cowan & Steel based on the morphological and physiological characteristics of bacterial cells. The mortality rate of bacteria was 30% in control, 44% in HCH and 51% in TPN plot respectively, in the process of obtaining pure culture. This suggests that the application of HCH or TPN enriched the fastidious bacteria in soil. The proportion of Gram-negative strains to the total isolates was 37% in control, 37% in HCH and 75% in TPN plot respectively. This means that the application of TPN enriched Gram-negative strains in soil. And the application of TPN increased the number of Gram-negative, nonspore-forming strains, and meanwhile decreased the number of spore-forming strains. In the results, the application of HCH or TPN changed considerably the composition of soil bacterial flora. And the influences of HCH and TPH on changes of the composition of soil bacterial flora were not equal each to each.