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Isolation of Bacillus subtilis GS-2 Producing γ-PGA from Ghungkukjang Bean Paste and Identification of γ-PGA (청국장으로부터 분리한 Poly(γ-glutamic acid)를 생산하는 균주 Bacillus subtilis GS-2의 분리 및 γ-PGA의 확인)

  • Bang, Byung-Ho;Jeong, Eun-Ja;Rhee, Moon-Soo;Kim, Yong-Min;Yi, Dong-Heui
    • Journal of Applied Biological Chemistry
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    • v.54 no.1
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    • pp.1-6
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    • 2011
  • ${\gamma}$-PGA(poly-${\gamma}$-glutamic acid) is an unusual anionic polypeptide that is made of D- and L-glutamic acid units connected by amide linkages between ${\alpha}$-amino and ${\gamma}$-carboxylic acid groups. ${\gamma}$-PGA has been isolated from many kinds of organisms. Many Bacillus strains produce ${\gamma}$-PGA as a capsular material of an extracellular viscous material. It is safe for eating as a viscosity element of fermented soybean products such as Chungkookjang and Natto. It is biodegradable, edible and nontoxic toward humans and the environment and its molecular weight varies from ten thousand to several hundred thousand depending on the kinds of strains used. Therefore, potential applications of ${\gamma}$-PGA and its derivatives have been of interest in the past few years in a broad range of industrial fields such as food, cosmetics, medicine, water-treatment, etc. In this study, a bacterium, Bacillus subtilis GS-2 isolated from the Korean traditional seasoning food, Chungkookjang could produce a large amount of ${\gamma}$-PGA with high productivity and had a simple nutrient requirement. Based on carbon utilization pattern and partial 16S rRNA sequence analysis, the GS-2 strain was identified as B. subtilis. The determination of purified ${\gamma}$-PGA was confirmed with thin layer chromatography (TLC), high performance liquid chromatography (HPLC), fourier transform infrared (FT-IR) spectra, and $^1H$-nuclear magnetic resonance ($^1H$-NMR) spectroscopy.

Effect of Ammonium Thiosulfate on Biological Activity in a Paddy Soil (Ammonium Thisoulfate 처리가 담수(湛水) 답토양(畓土壤)의 생물활성(生物活性)에 미치는 영향)

  • Lim, Sun-Uk;Seo, Young-Ho
    • Korean Journal of Soil Science and Fertilizer
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    • v.27 no.1
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    • pp.40-47
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    • 1994
  • The objective of this research was to examine the effect of ammonium thiosulfate(ATS) on urease activity and on biological and chemical properties of flooded paddy soil especially having high organic matter content by comparing with the effect of sodium thiosulfate(STS). The results obtained are summarized as follows. 1. The hydrolysis of urea was inhibited at 3 and 5 days after treatment of thiosulfate(ATS and STS) +glucose and thiosulfate only, respectively. The inhibitory effect of ATS on urea hydrolysis was slightly lower than that of STS in glusoce-added soils, but when the glucose was not added, the effects of ATS and STS were not different significantly. 2. The soil pH and Eh was lowered by 0.3~0.5 units and 30~120 mV, respectively, when incubated flooded soil with ATS and glucose at $25^{\circ}C$. 3. Soil respiration rate in/flooded soil was increased by 10~70% with the treatment of ATS during the 20 day experimental period. 4. The contents of acetic and butyric acid in thiosulfate treatment soil was below $10{\mu}g/g$, which was lower than that($220.3{\mu}g/g$) of critical growth inhibition of rice.

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Development of Benthic Macroinvertebrates Index (BMI) for Biological Assessment on Stream Environment (하천환경의 생물학적 평가를 위한 저서동물지수(BMI)의 개발)

  • Kong, Dongsoo;Son, Se-Hwan;Hwang, Soon-Jin;Won, Doo Hee;Kim, Myoung Chul;Park, Jung Ho;Jeon, Te Su;Lee, Jong Eun;Kim, Jong Hyun;Kim, Jong Sun;Park, Jaeheung;Kwak, Inn Sil;Ham, Sun Ah;Jun, Yung-Chul;Park, Young-Seuk;Lee, Jae-Kwan;Lee, Su-Woong;Park, Chang-Hee;Moon, Jeong-Suk;Kim, Jin-Young;Park, Hae Kyung;Park, Sun Jin;Kwon, Yongju;Kim, Piljae;Kim, Ah Reum
    • Journal of Korean Society on Water Environment
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    • v.34 no.2
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    • pp.183-201
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    • 2018
  • The tolerance of Korean benthic macroinvertebrates to organic pollution has been analyzed since the early 1990s. However, considering the fact that there have been related studies carried out in some European countries since the early 20th century, the history of the research in Korea is very short and there is still much knowledge to supplement. We revised the saprobic valency, the saprobic value and the indicator weight value of 190 benthic macroinvertebrates taxa through the data of water quality and individual abundance collected from 7,086 sampling units in Korea from 2008 to 2014. The individual abundance of Uracanthella (Ephemeroptera) as a representative, one of the most common and abundant taxa in Korea, showed a typical lognormal distribution to 5-day biochemical oxygen demand (BOD5) concentration, and a normal distribution to the class interval of BOD5 concentration according to saprobic series. The value combining the mean individual abundance and the relative frequency of occurrence was a more efficient indicator value than that of each property alone. Benthic Macroinertebrates Index (BMI) was newly proposed as a modification of the saprobic index of Zelinka and Marvan (1961). BMI showed extremely significant correlation (determination coefficient $r^2$ > 0.6, n = 569 sites) with the concentration of BOD5, and the coefficient was a little higher than those of the previous indices. Until now, there has been very little research on the assessment of biological integrity of benthic macroinvertebrates community in Korea. While continuing researches into improve the reliability of BMI, it is necessary to develop multimetric indices for evaluating the integrity, including the composition of species and functional guilds, and the richness and diversity of the community.

Rhizospheric fungi of Panax notoginseng: diversity and antagonism to host phytopathogens

  • Miao, Cui-Ping;Mi, Qi-Li;Qiao, Xin-Guo;Zheng, You-Kun;Chen, You-Wei;Xu, Li-Hua;Guan, Hui-Lin;Zhao, Li-Xing
    • Journal of Ginseng Research
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    • v.40 no.2
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    • pp.127-134
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    • 2016
  • Background: Rhizospheric fungi play an essential role in the plantesoil ecosystem, affecting plant growth and health. In this study, we evaluated the fungal diversity in the rhizosphere soil of 2-yr-old healthy Panax notoginseng cultivated in Wenshan, China. Methods: Culture-independent Illumina MiSeq and culture-dependent techniques, combining molecular and morphological characteristics, were used to analyze the rhizospheric fungal diversity. A diffusion test was used to challenge the phytopathogens of P. notoginseng. Results: A total of 16,130 paired-end reads of the nuclear ribosomal internal transcribed spacer 2 were generated and clustered into 860 operational taxonomic units at 97% sequence similarity. All the operational taxonomic units were assigned to five phyla and 79 genera. Zygomycota (46.2%) and Ascomycota (37.8%) were the dominant taxa; Mortierella and unclassified Mortierellales accounted for a large proportion (44.9%) at genus level. The relative abundance of Fusarium and Phoma sequenceswas high, accounting for 12.9% and 5.5%, respectively. In total,113 fungal isolates were isolated from rhizosphere soil. They were assigned to five classes, eight orders (except for an Incertae sedis), 26 genera, and 43 species based on morphological characteristics and phylogenetic analysis of the internal transcribed spacer. Fusarium was the most isolated genus with six species (24 isolates, 21.2%). The abundance of Phoma was also relatively high (8.0%). Thirteen isolates displayed antimicrobial activity against at least one test fungus. Conclusion: Our results suggest that diverse fungi including potential pathogenic ones exist in the rhizosphere soil of 2-yr-old P. notoginseng and that antagonistic isolates may be useful for biological control of pathogens.

Application of Toxicity Identification Evaluation Procedures for Toxic Effluents from the Aluminum Rolling Industry (알루미늄 가공 공장 배출 방류수의 독성 원인물질 탐색)

  • Ra, Jin-Sung;Lee, Jiho;Kim, Ki-Tae
    • Journal of Environmental Health Sciences
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    • v.41 no.5
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    • pp.305-313
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    • 2015
  • Objectives: The objective of this study is to identify toxicants causing acute toxicity in effluents from the aluminum rolling industry that violate the discharge limits in Korea. Methods: Whole effluent toxicity tests (WET) were conducted on effluent discharged from the aluminum rolling industry following the US EPA WET test methods. We collected effluent samples three times and evaluated acute toxicity by using Daphnia magna. We employed toxicity identification evaluation (TIE) procedures to identify toxicants causing toxicity in the effluent. Results: No specific chemical groups were identified in the seven different manipulations applied to the of wastewater effluent samples showing 1.3 toxic units (TU) according to the TIE phase I procedures. Water quality parameters for water hardness, electric conductivity and heavy metals (Mn) were 4,322 mg/l as $CaCO_3$, 11.39 mS/cm, and $5,551{\mu}g/l$, respectively. Considering water hardness and reference toxicity, high concentrations of Mn can be disqualified from the causative toxicants. Consequently, high ionic concentrations of $Na^+$(1,648 mg/l), $Ca^{2+}$(1,048 mg/l), $Mg^{2+}$(1,428 mg/l) and $SO_4{^{2-}}$(7,472 mg/l) were identified to be causative toxicants. Water hardness and electric conductivity exceed the $EC_{50}$ value obtained by biological toxicity tests using Daphnia magna. Conclusion: According to TIE procedures, high salt concentration is determined to be a major toxicant in the effluent of agro-industrial wastewater treatment plants receiving wastewater from the aluminum rolling industry.

More about Taxonomic Sufficiency: A Case Study using Polychaete Communities in a Subtropical Bay Moderately Affected by Urban Sewage

  • Muniz Pablo;Pires-Vanin Ana M. S.
    • Ocean Science Journal
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    • v.40 no.3
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    • pp.127-143
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    • 2005
  • The taxonomic sufficiency approach has been proposed as a surrogate for the typical analysis of species-abundance data, especially in conditions involving prominent pollution gradients. Here, we evaluate the use of taxonomic sufficiency with infralittoral macrobenthic data derived from samples taken in a moderate polluted subtropical environment in southeastern Brazil, analysing five taxonomic levels and including two functional levels throughout polychaete feeding guilds and trophic groups. The data were collected seasonally at nine stations and studied for two abundance data series (0.5 and 1.0 mm sieve mesh-size). The results showed a similar ordination pattern between the two sieve mesh-size, but with the 0.5 mm sieve data a different pattern was observed during austral summer. A slight loss of information was detected using genus, family, polychaete species and their feeding guilds as taxonomic/functional units. These results together with those of the cost! benefit ratio, suggested that the family level seemed to be sufficient to detect the impact caused by moderate pollution in this shallow-water, subtropical environment. In additional, through the use of feeding guilds, similar patterns are obtained. Correlation analysis showed that chlorophyll a, total organic matter, zinc, and chromium sediment content were the variables that best explained the biological pattern observed and not always the best correlation coefficient occurring at the species level. The feeding guild approach seems to be useful and generates interpretable results similar to those obtained with the species level of the whole macroinfauna. The results showed an important cost reduction in the sample processing, suggesting that it is possible to adopt a coarser taxonomic level monitoring program even in species-rich communities.

Isolation and identification of canine adenovirus type 2 from a naturally infected dog in Korea

  • Yang, Dong-Kun;Kim, Ha-Hyun;Yoon, Soon-Seek;Lee, Hyunkyoung;Cho, In-Soo
    • Korean Journal of Veterinary Research
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    • v.58 no.4
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    • pp.177-182
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    • 2018
  • Canine adenovirus type 2 (CAV-2) infection results in significant respiratory illness in dogs. Isolating and culturing CAV-2 allows for investigations into its pathogenesis and the development of vaccines and diagnostic assays. In this study, we successfully isolated a virus from a naturally infected dog in Gyeonggi-do, Korea. The virus was propagated in Madin-Darby canine kidney (MDCK) and Vero cells and showed a specific cytopathic morphology that appeared similar to a bunch of grapes. The virus was first confirmed as CAV-2 based on these cytopathic effects, an immunofluorescence assay, hemagglutination assay, and electron microscopy. The viral titer of the isolate designated APQA1601 reached $10^{6.5}$ 50% tissue culture infections dose per mL in MDCK cells and exhibited no hemagglutination units with erythrocytes from guinea pig. The virus was also confirmed by polymerase chain reaction and next-generation sequencing. The APQA1601 strain had the highest similarity (~99.9%) with the Toronto A26/61 strain, which was isolated in Canada in 1976 when the nucleotide sequences of the full genome of the APQA1601 strain were compared with those of other CAV strains. Isolating CAV-2 will help elucidate the biological properties of CAV-2 circulating in Korean dogs.

Isolation and identification of mammalian orthoreovirus type 3 from a Korean roe deer (Capreolus pygargus)

  • Yang, Dong-Kun;An, Sungjun;Park, Yeseul;Yoo, Jae Young;Park, Yu-Ri;Park, Jungwon;Kim, Jong-Taek;Ahn, Sangjin;Hyun, Bang-Hun
    • Korean Journal of Veterinary Research
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    • v.61 no.2
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    • pp.13.1-13.8
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    • 2021
  • Mammalian reovirus (MRV) causes respiratory and intestinal disease in mammals. Although MRV isolates have been reported to circulate in several animals, there are no reports on Korean MRV isolates from wildlife. We investigated the biological and molecular characteristics of Korean MRV isolates based on the nucleotide sequence of the segment 1 gene. In total, 144 swabs from wild animals were prepared for virus isolation. Based on virus isolation with specific cytopathic effects, indirect fluorescence assays, electron microscopy, and reverse transcription-polymerase chain reaction, only one isolate was confirmed to be MRV from a Korean roe deer (Capreolus pygargus). The isolate exhibited a hemagglutination activity level of 16 units with pig erythrocytes and had a maximum viral titer of 105.7 50% tissue culture infectious dose (TCID50)/mL in Vero cells at 5 days after inoculation. The nucleotide and amino-acid sequences of the partial segment S1 of the MReo2045 isolate were determined and compared with those of other MRV strains. The MReo2045 isolate had nucleotide sequences similar to MRV-3 and was most similar (96.1%) to the T3/Bat/Germany/342/08 strain, which was isolated in Germany in 2008. The MReo2045 isolate will be useful as an antigen for sero-epidemiological studies and developing diagnostic tools.

Physico-chemical Characteristics of Yogurt by Lactobacillus spp. from Pickles (젓갈에서 분리한 Lactobacillus spp.로 제조한 요구르트의 이화학적 특성)

  • Rhee, Young-Hwan;Na, Han-Ju;Lee, Yong-Kyu;Shin, Seung-Yee;Kim, Jong-Hyun
    • Applied Biological Chemistry
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    • v.40 no.1
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    • pp.12-17
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    • 1997
  • Three strains of lactic acid bacteria were isolated from fish and shrimp pickles. Two strains were identified as Lactobacillus casei, and one strain as L. Pentosus, respectively. All three strains were used as starters in producing yogurts. The physico-chemical characteristics of yogurts were examined. The original pH, titratable acidity, visicosity and viable cell counts of the yogurts were $4.03{\sim}4.26,\;1.049{\sim}l.217%,\;1,772{\sim}2,232\;cps\;and\;1.4{\times}10^9{\sim}1.6{\times}10^9\;cfu/ml$, respectively. In evaluating buffer capacity, $12.50{\sim}14.06\;ml$ 1.0N HCl was consumed to titrate 100 ml of yogurt to pH value 2 units below the original pH value and $9.46{\sim}13.06\;ml$ of 1.0N NaOH was consumed to pH value 4 units above the original pH value. The ${\beta}-galactosidase$ activity reached maximum at 48 hrs, and reduced gradually during fermantation. After 2 hr incubation of yogurts at $37^{\circ}C$ under different pH conditions, ${\beta}-galactosidase$ activities of three strains were reduced to 50% at pH 3.5, but there were no remaining activities neither at pH 2.5 nor at pH 1.5. Under the same pH conditions the number of viable cells decreased to $1.9{\times}10^6{\sim}1.8{\times}10^8\;cfu/ml$ at pH 2.5 and $1.0{\times}10^3{\sim}2.4{\times}10^5$ at pH 1.5, respectively. However, no significant difference was found at pH 3.5.

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Soybean meal substitution by dehulled lupine (Lupinus angustifolius) with enzymes in broiler diets

  • Mera-Zuniga, Fredy;Pro-Martinez, Arturo;Zamora-Natera, Juan F;Sosa-Montes, Eliseo;Guerrero-Rodriguez, Juan D;Mendoza-Pedroza, Sergio I;Cuca-Garcia, Juan M;Lopez-Romero, Rosa M;Chan-Diaz, David;Becerril-Perez, Carlos M;Vargas-Galicia, Artemio J;Bautista-Ortega, Jaime
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.4
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    • pp.564-573
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    • 2019
  • Objective: Evaluate the effects of i) dehulling of lupine seed on chemical composition and apparent metabolizable energy (AME) and ii) soybean meal substitution by dehulled lupine seed in broiler diets with enzymes on productive performance, size of digestive organs and welfare-related variables. Methods: Experiment 1, chemical composition and AME were determined in whole and dehulled lupine seed. Experiment 2, two hundred eighty-eight one-day-old male Ross 308 broilers were used. The experimental diets were maize-soybean meal (MS), MS with enzymes (MSE) and maize-dehulled lupine seed with enzymes (MLE). Diets were assigned to the experimental units under a completely randomized design (eight replicates per diet). The body weight (BW) gain, feed intake, feed conversion, digestive organ weights, gait score, latency to lie down and valgus/varus angulation were evaluated. Results: The dehulling process increased protein (25.0% to 31.1%), AME (5.9 to 8.8 MJ/kg) and amino acid contents. The BW gain of broilers fed the MLE diet was similar (p>0.05) to that of those fed the MS diet, but lower than that of those fed the MSE diet. Feed intake of broilers fed the MLE diet was higher (p<0.05) than that of those fed the MS diet and similar (p>0.05) to those fed the MSE diet. Feed conversion of broilers fed the MLE diet was 8.0% and 8.7% higher (p<0.05) than that of those fed the MS and MSE diets, respectively. Broilers fed the MLE diet had the highest (p<0.05) relative proventriculus and gizzard weights, but had poor welfare-related variables. Conclusion: It is possible to substitute soybean meal by dehulled lupine seed with enzymes in broiler diets, obtaining similar BW gains in broilers fed the MLE and MS diets; however, a higher feed intake is required. Additionally, the MLE diet reduced welfare-related variables.