• Asian-Australasian Journal of Animal Sciences
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• v.30 no.2
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• pp.171-180
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• 2017

Objective: This study investigated the association of enzyme-producing microbes and their enzymes with starch and hemicellulose degradation during fermentation of total mixed ration (TMR) silage. Methods: The TMRs were prepared with soybean curd residue, alfalfa hay (ATMR) or Leymus chinensis hay (LTMR), corn meal, soybean meal, vitamin-mineral supplements, and salt at a ratio of 25:40:30:4:0.5:0.5 on a dry matter basis. Laboratory-scale bag silos were randomly opened after 1, 3, 7, 14, 28, and 56 days of ensiling and subjected to analyses of fermentation quality, carbohydrates loss, microbial amylase and hemicellulase activities, succession of dominant amylolytic or hemicellulolytic microbes, and their microbial and enzymatic properties. Results: Both ATMR and LTMR silages were well preserved, with low pH and high lactic acid concentrations. In addition to the substantial loss of water soluble carbohydrates, loss of starch and hemicellulose was also observed in both TMR silages with prolonged ensiling. The microbial amylase activity remained detectable throughout the ensiling in both TMR silages, whereas the microbial hemicellulase activity progressively decreased until it was inactive at day 14 post-ensiling in both TMR silages. During the early stage of fermentation, the main amylase-producing microbes were Bacillus amyloliquefaciens (B. amyloliquefaciens), B. cereus, B. licheniformis, and B. subtilis in ATMR silage and B. flexus, B. licheniformis, and Paenibacillus xylanexedens (P. xylanexedens) in LTMR silage, whereas Enterococcus faecium was closely associated with starch hydrolysis at the later stage of fermentation in both TMR silages. B. amyloliquefaciens, B. licheniformis, and B. subtilis and B. licheniformis, B. pumilus, and P. xylanexedens were the main source of microbial hemicellulase during the early stage of fermentation in ATMR and LTMR silages, respectively. Conclusion: The microbial amylase contributes to starch hydrolysis during the ensiling process in both TMR silages, whereas the microbial hemicellulase participates in the hemicellulose degradation only at the early stage of ensiling.

• Research in Plant Disease
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• v.9 no.4
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• pp.229-236
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• 2003

The 1080 epiphytic bacteria obtained from 370 samples of pome and stone fruits including apple, pear, peach, grape, apricot and Chinese quince were screened for antagonistic activity against postharvest pathogens, Penicillium expansum, Alternaria alternata and Botrytis cinerea. Among tested antagonistic bacteria, eight bacterial isolates inhibited mycelial growth of the postharvest pathogens and were identified as Bacillus amyloliquefaciens (three strains), B. megaterium, B. subtilis var. gladioli, B. licheniformis, B. pumilus and Serratia marcescens based on biochemical characteristics and utility of carbon and nitrogen compounds (Biolog system). Eight carbohydrates were evaluated for their effect on mycelial growth and germination of the postharvest pathogen, P. expansum to select nutrients for enhancing bio-control efficacy. The growth of four selected antagonists, B. amyloliquefaciens P43-2, B. amyloliquefaciens A71-2, B. licheniformis P94-1, and S. marcescens P76-9 were also tested. As a result, 1% glucose (w/v) strongly stimulated growth of the antagonists, suppressed mycelial growth of the postharvest pathogen, and had a little comparatively stimulatory effect on germination of the the postharvest pathogen. It was confirmed that the addition of 1% glucose (w/v) greatly enhanced biocontrol effect of B. amyloliquefaciens P43-2, B. licheniformis P94-1, and S. marcescens P76-9. Application of B. amyloliquefaciens P43-2, B. licheniformis P94-1, and S. marcescens P76-9 with the addition of 1% glucose (w/v) increased the control efficacy up to 48%, 46%, 14% compared with those of the antagonists without glucose, respectively. When the antagonists were applied to control postharvest disease caused by P. expansum in apple wounds, the population of B. amyloliquefaciens P43-2 and B. licheniformis P94-1 increased until 4 days after inoculation (DAI) of the antagonists and then decreased from 10 DAI. Meanwhile the population of S. marcescens P76-9 decreased at early stage (4 DAI), but increased from 7 DAI, and finally maintained constantly until 10 DAI in apple wounds.

• Applied Biological Chemistry
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• v.7
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• pp.53-58
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• 1966

A study was carried out to investigate the microorganisms and their floral changes during the tobacco fermentation. The results were sumarized as follows. I. The molds in the "tobacco leaves" were isolated and identified as follows; Aspergllus flavus Link, A. restrictus G. Smith, A. nidulans (Eidam) Wint, A. awamori, Oidium sp. Edmundmasonia sp. Spicaria sp. II. The bacteria in the tobacco were isolated and identified as follows; Bacillus subtilis, B. subtilis var aterriums. B. licheniformis, B. cereus, B. Pumilus, B. megaterium, Flavobacterium harrisonii, Aerobacter aerogenes. III. The counts of the microorganisms on leaves taken from bulks of the fermenting leaf tobacco revealed the presence of relatively small number on the initial stage of the fermentation. During the tobacco fermentation the number of molds increase gradually to the maximum until the 14 th. day of the fermenation, followed by showing, the plateau, and the bacteria population revealed the maximum on the 7 th. day, then declined slowly.

• Korean Journal of Microbiology
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• v.48 no.2
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• pp.134-140
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• 2012

Asian dust storms originating in the arid desert of China and Mongolia usually occur from late winter through spring, and more than one million tons of dust per year is transported to the Korean Peninsula by the prevalent westerly winds. We supposed that these dust particles could include bioaerosols and act as carriers of microorganisms. In order to clarify the dynamics of microorganisms moving with these particles, the concentration and composition of microorganisms associated with settled particles were compared between samples collected during Asian dust events and those under non-dust periods. From February to April 2008, settled dust particles were collected at one location in Ulsan using rainfall meter of 200 mm diameter. During this period, there was one Asian dust event in Ulsan. The bacterial concentrations were higher in samples collected during Asian dust event than those under non-dust period, whereas fungal concentrations were rather similar regardless of the Asian dust event. We analyzed 16S rRNA gene sequences of 45 bacterial isolates obtained from the settled particle samples. These isolates belonged to either genus Bacillus or genus Streptococcus and were tentatively identified as B. amyloliquefaciens, B. aryabhattai, B. atrophaeus, B. licheniformis, B. megaterium, B. methylotrophicus, B. pumilus, B. sonorensis, B. subtlis, B. vallismortis, S. epidermidis, and S. succinus. In cases of fungal isolates, genera such as Mucor, Alternaria, Cladosporium, and Aspergillus were tentatively identified from samples collected at both Asian dust and non-Asian dust periods. It appears that endospore-forming bacteria such as Bacillus sp. rather than fungal spores are more likely to be associated with Asian dust particles.

• Journal of Environmental Science International
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• v.16 no.9
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• pp.1103-1109
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• 2007

The aim of this study was to isolate mesophilic chicken feather-degrading bacteria and to evaluate feather hydrolysate as alternative biofertilizer. Isolate RS7 was isolated from compost and identified as Bacillus pumilus according to API analysis and l6S rDNA sequencing analysis. Chicken feathers were completely degraded after 5 days of cultivation at $30^{\circ}C$. Feather hydrolysate treated by B. pumilius RS7 positively influenced Helianthus sannuus L. (sunflower) growth (e.g. growth rate, number and dry weight of leave, and flowering rate). These results suggest that feather hydrolysate prepared using B. pumilius RS7 could successfully be used as alternative biofertilizer, thereby reducing the environmental impact of feather waste from the poultry industry.

• Journal of Microbiology and Biotechnology
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• v.8 no.1
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• pp.28-33
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• 1998

A gene coding for ${\beta}$-xylosidase from thermophilic xylanolytic Bacillus sp. KK-1 was cloned into Escherichia coli using plasmid pBR322. Recombinant plasmid DNAs were isloated from E. coli clones which were capable of hydrolyzing 4-methylumbelliferyl-${\beta}$-D xylopyranoside. Restriction analysis showed the DNAs to share a common insert DNA. Xylo-oligosaccharides, including xylotriose, xylotetraose, xylopentaose, and xylobiose were hydrolyzed to form xylose as an end product by cell-free extracts of the E. coli clones, confirming that the cloned gene from strain KK-1 is ${\beta}$-xylosidase gene. The ${\beta}$-xylosidase gene of strain KK-1 designated as xylB was completely sequenced. The xylB gene consisted of an open reading frame of 1,602 nucleotides encoding a polypeptide of 533 amino acid residues, and a TGA stop codon. The 3' flanking region contained one stem-loop structure which may be involved in transcriptional termination. The deduced amino acid sequence of the KK-1 ${\beta}$-xylosidase was highly homologous to the ${\beta}$-xylosidases of Bacillus subtilis and Bacillus pumilus, but it showed no similarity to a thermostable ${\beta}$-xylosidase from Bacillus stearothermophilus.

• BMB Reports
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• v.44 no.3
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• pp.193-198
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• 2011

The chitinase-producing strain SC081 was isolated from Korean traditional soy sauce and identified as Bacillus atrophaeus based on a phylogenetic analysis of the 16S rDNA sequence and a phenotypic analysis. A gene encoding chitinase from B. atrophaeus SC081 was cloned in Escherichia coli and was named SCChi-1 (GQ360078). The SCChi-1 nucleotide sequences were composed of 1788 base pairs and 596 amino acids, which were 92.6, 89.6, 89.3, and 78.9% identical to those of Bacillus subtilis (ABG57262), Bacillus pumilus (ABI15082), Bacillus amyloliquefaciens (ABO15008), and Bacillus licheniformis (ACF40833), respectively. A recombinant SCChi-1 containing a hexahistidine tag at the amino-terminus was constructed, overexpressed, and purified in E. coli to characterize SCChi-1. $H_6SCChi$-1 revealed a hydrolytic band on zymograms containing 0.1% glycol chitin and showed the highest lytic activity on colloidal chitin and acidic chitosan. The optimal temperature and pH for chitinolytic activity were $50^{\circ}C$ and pH 8.0, respectively.

• The Journal of Engineering Geology
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• v.23 no.1
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• pp.37-46
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• 2013

Leakage of leachate from animal carcass disposal is a significant issue because disease can easily spread to humans and other livestock. In this study, we analyzed the physicochemical properties of leachate and tested for the presence of aerobic bacteria in leachate using molecular biology methods, for 16 animal carcass disposals in the first stage (after burial for 5 months). Leachate physicochemical analysis revealed higher total coliforms, TOC, $NH^{4+}$, and $NO^{3-}$ concentrations compared with previously published data. In most leachate samples, the concentrations of $NH^{4+}$ and $NO^{3-}$ exceeded the Korean guideline values for drinking water. In 16S rRNA sequence analysis of the distribution of leachate under aerobic conditions, Bacillus pumilus, Lysinibacillus sphaericus, and B. sphaericus were observed with high frequency, whereas no food-poisoning-related bacteria such as B. cereus or Salmonella were detected. The present findings improve our knowledge of the transport of leachate from animal carcass disposal sites through geologic media, and are useful in risk analysis and for subsequent studies.

• Journal of Microbiology and Biotechnology
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• v.22 no.4
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• pp.437-447
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• 2012

Natural and beneficial associations between plants and bacteria have demonstrated potential commercial application for several agricultural crops. The sunflower has acquired increasing importance in Brazilian agribusiness owing to its agronomic characteristics such as the tolerance to edaphoclimatic variations, resistance to pests and diseases, and adaptation to the implements commonly used for maize and soybean, as well as the versatility of the products and by-products obtained from its cultivation. A study of the cultivable bacteria associated with two sunflower cultivars, using classical microbiological methods, successfully obtained isolates from different plant tissues (roots, stems, florets, and rhizosphere). Out of 57 plant-growth-promoting isolates obtained, 45 were identified at the genus level and phylogenetically positioned based on 16S rRNA gene sequencing: 42 Bacillus (B. subtilis, B. cereus, B. thuringiensis, B. pumilus, B. megaterium, and Bacillus sp.) and 3 Methylobacterium komagatae. Random amplified polymorphic DNA (RAPD) analysis showed a broad diversity among the Bacillus isolates, which clustered into 2 groups with 75% similarity and 13 subgroups with 85% similarity, suggesting that the genetic distance correlated with the source of isolation. The isolates were also analyzed for certain growth-promoting activities. Auxin synthesis was widely distributed among the isolates, with values ranging from 93.34 to 1653.37 ${\mu}M$ auxin per ${\mu}g$ of protein. The phosphate solubilization index ranged from 1.25 to 3.89, and siderophore index varied from 1.15 to 5.25. From a total of 57 isolates, 3 showed an ability to biologically fix atmospheric nitrogen, and 7 showed antagonism against the pathogen Sclerotinia sclerotiorum. The results of biochemical characterization allowed identification of potential candidates for the development of biofertilizers targeted to the sunflower crop.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.7
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• pp.1035-1040
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• 2016

This study investigated the prevalence of foodborne pathogens and the bacterial community of traditional Doenjang collected from a longevity area in Korea as well as the antagonistic effect of traditional Doenjang isolates against Bacillus cereus to estimate the microbiological safety of traditional Doenjang. Aerobic bacteria showed $10^6{\sim}10^9CFU/g$, whereas coliform bacteria was not detected. Foodborne pathogens were not detected except B. cereus, which was detected in seven samples out of 10 Doenjang samples. A total of 327 isolates were identified from traditional Doenjang. The isolates consisted of Bacillus subtilis 155 (47.4%), Bacillus licheniformis 68 (20.8%), Bacillus amyloliquefaciens 46 (14.1%), and Bacillus pumilus 18 (5.5%). Antagonistic effect against B. cereus was detected in 20 (6.1%) of 327 isolates, which consisted of B. subtilis (12 strains), B. amyloliquefaciens (5 strains), and B. licheniformis (3 strains). The inhibitory zone for the antagonistic effect was 9.0~12.0 mm in diameter. Although a small amount of traditional Doenjang was tested in this study, these results indicated that the potential risk of B. cereus in traditional Doenjang is lower than generally presumed. It is necessary to monitor the antagonistic effect of traditional Doenjang isolates against B. cereus.