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Genetic Characteristics and Virulence Factors of Pandemic Vibrio parahaemolyticus Isolated in South Korea and Japan (한국과 일본에서 유행하는 장염비브리오의 병원성 인자와 유전자의 특성)

  • Hong, Seuk-Won;Moon, Ji-Young;Lee, Bok-Kwon;Kim, Yung-Bu
    • Journal of Life Science
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    • v.17 no.3 s.83
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    • pp.386-395
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    • 2007
  • A total of 27 strains of Vibrio parahaemolyticus (18 strains isolated from Korea and 9 strains from Japan) were serotyped and examined for biochemical characteristics, antimicrobial susceptibility patterns, cytotoxicity assay, thermostable direct hemolysin (TDH) production and molecular epidemiology. Using polymerase chain reaction (PCR) method and DNA probe hybridization method, the strains were tested for toxR, tdh, trh and ORF 8 genes. The V. parahaemolyticus isolated from patients were belonged to 8 different serotypes : O3:K6, O1:K38, O3:K57, O4:K9, O4:Kl2, O4:K68, O5:Kl5 and O6:K46. Urease-positive strain possessed the trh gene, and conversely, urease-negative strains lacked the gene, indicating that urease production by V. parahemolyticus strains strongly correlates with the possession of the trh gene. Most strains showed multiple resistant to more than three antibiotics and the antibiogram could be classified into 6 group (I to VI). All of the O3:K6 strains isolated in South Korea and Japan producted TDH at high levels. The TDH titers ranged between 256 and 2.048, and the average titer was 1009. To distinguish the new and increasingly common V. parahaemolyticus strains from clinical isolates, ORF 8 is a useful genetic marker. After Southern hybridization, the HindIII restriction fragment patterns of the tdh gene were grouped one type, respectively. One type showed two bands one of which was 4.3kb and the other was 11.5kb in size. Variation between the O3:K6 serotype are minor when compared to the differences seen with the non O3:K6 strains. The migration patterns of Not I -digested of the total DNA of the O3:K6 strains were similar, and only slight variations were observed between the serotypes. By contrast, the O3:K6 strains and non O3:K6 had markedly different profiles. In conclusion, Random amplified polymorphic DNA (RAPD) profile using appropriate primers was an effective epidemiological marker.

Isolation and Characteristics of Yersinia spp. from Mineral Spring Waters (먹는 샘물에서 분리한 Yersinia spp.의 분리 및 특성)

  • 이택수;박부길;오덕환
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.5
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    • pp.796-801
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    • 2001
  • A total of 277 mineral spring water samples in Kangwon province from 1999 to 2000 were analyzed for the presence of Yersinia spp. by the conventional Food and Drug Administration protocol, and presumptive strains were identified by morphological, cultural and biochemical tests according to Bergey’s manual. Also, the biotypes, serotypes, and susceptibility to 12 antibiotics were tested. Among the total 277 mineral spring water samples, 40 samples (14.4%) were found to be contaminated with Yersinia species. Among the 40 strains of Yersinia spp. isolates, 33 strains (82.5%) for Yersinia enterocolitica, 4 strains (10%) for Yersinia frederiksenii, 2 strains (5%) for Yersinia intermedia, and 1 strain (2.5%) for Yersinia sakazaki were identified, respectively. Of 40 Yersinia spp. isolates, Yersinia enterocolitica (82.5%) was the most predominant species in the mineral spring water samples compared to other Yersinia species. Compared to direct culture method after KOH treatment and KOH treatment method after cold enrichment for better isolation ratio of according to comparision of Yersinia species, the detection ration (18.5%) of KOH treatment method after cold enrichment was about 3 times better than that (6.1%) of direct culture method after KOH treatment. According to serotypes of Y. enterocolitica isolates, O : 5 (12.9%) was the most predominant and followed by O : 3 (9.7%), O : 8 (6.5%), and O : 9 (3.2%), and others. For biotypes of Y. enterocolitica isolates, 1A (71.0%) was the most predominantly abundant and followed by 3A (12.9%), 3B (9.7%), 1B (3.2%) and 5 (3.2%). Also, an antibiotic susceptibility test showed that Yersinia spp. isolates were very susceptible to the antibiotics tested, but they were very strongly resistant to ampicillin, cephalothin and carbenicillin.

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Dietary effect of Bacillus subtilis MD-02 on Innate Immune Response and Disease Resistance in Olive Flounder, Paralichthys olivaceus (넙치(Paralichthys olivaceus)의 비특이적 면역반응 및 병 저항성에 대한 Bacillus subtilis MD-02의 효과)

  • Kim, Dong-Hwi;Heo, Moon-Soo
    • Microbiology and Biotechnology Letters
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    • v.47 no.1
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    • pp.132-138
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    • 2019
  • Among several marine-derived microorganisms isolated from the coast of Jeju Island that had antimicrobial activity against fish disease pathogens, Bacillus subtilis MD-02 was tested for its dietary effect on the innate immune response and disease resistance of olive flounder. Strain MD-02 was fed to the olive flounder at a concentration of $1.2{\times}10^4$, $1.2{\times}10^6$, or $1.2{\times}10^8CFU/100g$, respectively. Consequently, the hematocrit was higher in these three groups than that in the control group at 4 weeks, and the aspartate aminotransferase and alanine aminotransferase levels were decreased in the $1.2{\times}10^8$ and $1.2{\times}10^4CFU/100$ groups compared with the control group levels. The amylase activity and total protein were significantly increased in the $1.2{\times}10^4CFU/100g$ group at 3 weeks. The innate immune response, determined from the lysozyme and macrophage activities, was higher in the $1.2{\times}10^8CFU/100g$ group than in the control group. In addition, treatment of the olive flounders with Streptococcus parauberis at $1.2{\times}10^6CFU/ml$ confirmed the mortality rate, which was 100% in the control group and 40-60% in the groups fed B. subtilis MD-02, indicating that the fish had resistance to fish disease pathogens. Therefore, it was confirmed that when fed MD-02, olive flounder builds an innate immune response and acquires resistance to fish disease pathogens, indicating that B. subtilis MD-02 can be developed as a beneficial feed additive.

Evaluation of the Anti-oxidant Activity of Pueraria Extract Fermented by Lactobacillus rhamnosus BHN-LAB 76 (Lactobacillus rhamnosus BHN-LAB 76에 의한 Pueraria 발효 추출물의 항산화 활성 평가)

  • Kim, Byung-Hyuk;Jang, Jong-Ok;Lee, Jun-Hyeong;Park, Ye-Eun;Kim, Jung-Gyu;Yoon, Yeo-Cho;Jeong, Su Jin;Kwon, Gi-Seok;Lee, Jung-Bok
    • Journal of Life Science
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    • v.29 no.5
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    • pp.545-554
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    • 2019
  • The phytochemical compounds of Pueraria, a medicinally important leguminous plant, include various isoflavones that have weak estrogenic activity and a potential role in preventing chronic disease, cancer, osteoporosis, and postmenopausal syndrome. However, the major isoflavones are derivatives of puerarin and occur mainly as unabsorbable and biologically inactive glycosides. The bioavailability of the glucosides can be increased by hydrolysis of the sugar moiety using ${\beta}$-glucosidase. In this study, we investigated the antioxidant effects of a Pueraria extract after fermentation by Lactobacillus rhamnosus BHN-LAB 76. The L. rhamnosus BHN-LAB 76 strain was inoculated into Pueraria powder and fermented at $37^{\circ}C$ for 72 hr. The total polyphenol content of the Pueraria extract increased by about 134% and the total flavonoid content increased around 110% after fermentation with L. rhamnosus BHN-LAB 76 when compared to a non-fermented Pueraria extract. Superoxide dismutase-like activities, DPPH radical scavenging, and ABTS radical scavenging increased by approximately 213%, 190%, and 107%, respectively, in the fermented Pueraria extract compared to the non-fermented Pueraria extract. Fermentation of Pueraria extracts with L. rhamnosus BHN-LAB 76 is therefore possible and can effectively increase the antioxidant effects. These results can be applied to the development of improved foods and cosmetic materials.

Development of Genetic Selection Marker via Examination of Genome in Bacillus velezensis K10 (Bacillus velezensis K10 유전체 분석을 통한 균주 선발 마커 개발)

  • Sam Woong Kim;Young Jin Kim;Tae Wook Lee;Won-Jae Chi;Woo Young Bang;Tae Wan Kim;Kyu Ho Bang;Sang Wan Gal
    • Journal of Life Science
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    • v.33 no.11
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    • pp.897-904
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    • 2023
  • This study was done to develope genetic markers with the unique characteristics of genes according to the genomic information of Bacillus velezensis K10. B. velezensis K10 maintained a total of 4,159,835 bps, which was found to encode 5,136 open reading frames (orfs). B. velezensis K10 was found to have much more gene migration due to external factors overall compared to standard strain B. velezensis JS25R. In order to discover genetic selection markers, orfs on the genome to be easily induced to gene mutation were surveyed such as recombinase, integrase, transposase, and phage-related genes. As a result of the investigation, 9 candidate markers were isolated with high possibility as genetic selection markers. Although a part in the various origin's areas showed specificities in comparison with homology, the selected markers were all existed in phage-related areas because they were relatively lower homologies in phage-related genes. PCR analysis was done on B. licheniformis K12, B. velezensis K10, B. subtilis, and B. cereus to establish them as inter-species candidate selection markers. As a result, it was confirmed that B. velezensis K10-specific PCR products were formed in a total of 6 primer sets such as BV3 and BV5 to 9. On the other hand, analysis at the subspecies level observed the formation of B. velezensis K10-specific PCR products in 4 primer sets such as BV3, 5, 8, and 9. Among them, since BV5 and BV8 were detected by very specific results, we suggest that BV5 and 8 can be used as B. velezensis K10 gene selection markers at the species and sub-species level.

Study on the Genetic Variations of the Economic Traits by Backcrossing in Commercial Chickens (실용계군에 있어서 누진퇴교배에 의한 주요경제형질의 유전적 변이에 관한 연구)

  • 이종극;오봉국
    • Korean Journal of Poultry Science
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    • v.16 no.2
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    • pp.61-71
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    • 1989
  • The purposes of this study were to investigate the genetic variations by backcrossing in commercial chickens. Backcrossing was carried out successively back to parent stock (P.S). Heritabilities and genetic correlation coefficients were estimated to verify the genetic variations. The data obtained from a breeding programme with commercial chickens (I strain) were collected from 1955 to 1987 at Poultry Breeding Farm, Seoul National University. Data came from a total of 1230 female offspring. The results obtained are summarized as follows: 1. The general performance ($Mean\pmStandard deviation$) of each trait was $663.94\pm87.11$g for 8 weeks body weight, $1579.1\pm155.43$g for 20 weeks body weight, $2124.1\pm215.3$g for 40 weeks body weight, $2269.1\pm242.94$g for 60 weeks body weight, $168.43\pm12.94$ day for a9e at sexual maturity (SM), $214.52\pm29.82$ eggs , for total egg number to 60 weeks of age (TEN), $61.45\pm3.48$ g for average weight (AEW), $13180.7\pm1823.22$ g for total egg mass to 60 weeks of age(TEM). All traits, except 10 weeks body weight and AEW, were significant for the degrees of backcross (p<0.01). 2. The pooled estimates of heritabilities derived from the sire, dam and combined variance components were 0.47~0.52 for age at sexual maturity (SM), 0.07~0.37 for total egg number (TEN), 0.40~0.54 for average egg weight (AEW), 0.18~0.27 for total egg mass (TEM). High heritability estimates were found for SM and AEW. TEN and TEM were estimated to be a lowly heritable traits. Heritability estimates from dam components were higher than those from sire components. These differences might be due to non-additive genetic effect and maternal effect. 3. The estimates of heritabilities and standard errors derived from combined variance components for different degrees of backcross were $0.47\pm0.11$ (BCO), $0.42\pm0.16$ (BC1), $0.51\pm0.29$ (BC2) for TEN, $0.59\pm0.20$ (BCO), $0.43\pm0.17$ (BC1), $0.35\pm0.18$ (BC2) for AEW, $0.28\pm0.12$(BC0), $0.20\pm0.11$(BC1), $0.18\pm0.14$ (BC2) for TEM. Heritability estimates for AEW and TEM were decreased by backcrossing while those for SM and TEN remained constant. Since backcrossing contributes to increased homozygosity, the genetic variation of the traits (AEW and TEM) decreased . 4. The pooled estimates of genetic correlation coefficients were -0.55 between SM and TEN, 0.20 between SM and AEW, -0.29 between TEN and AEW, 0.82 between TEM and TEN, 0.31 between TEM and AEW, -0.42 between TEM and SM. The genetic correlation between TEM and TEN was higher than that between TEM and AEW, and it was suggested that egg mass was strongly affected by egg number. Also, age at sexual maturity(SM) contributes to egg mass(TEM). 5. When backcrossing was carried out successively, the genetic correlation between TEM and TEN increased (BC0:0.79, BC1:0.82, BC2:0.91) but those between TEM and SM decreased (BC0:-0.54, BC1:-0.36, BC2:-0.09) with successive backcrosses.

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Microbiological and Enzymological Studies on the Flavor Components of Sea Food Pickles (젓갈등속(等屬)의 정미성분(呈味成分)에 관(關)한 미생물학적(微生物學的) 및 효소학적(酵素學的) 연구(硏究))

  • Lee, Ke-Ho
    • Applied Biological Chemistry
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    • v.11
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    • pp.1-27
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    • 1969
  • More than thirty kinds of sea food pickles have been eaten in Korea. Out of these salted yellow tail pickle, salted clam pickle, salted oyster pickle, and salted cuttlefish pickle were employed for the analysis of their components, identification of main fermenting microbes, and determination of enzyme characteristics concerned. Also studied was the effect of enzymic action of microbes, which are concerned with the fermenting of pickles, on the production of flavorous 5'-mononucleotides and amino acids. The results are summarized as follows: 1. Microflora observed in the pickles are: (a) Total count of viable cells after 1-2 months of pickling was found to be $10^7$ and that after 6 months decreased to $10^4$. (b) Microbial occurence in the early stage of pickling was observed to be 10-20% Micrococcus spp., 10-20% Brevibacterium spp., 0-30% Sarcina spp., 20-30% Leuconostoc spp., ca 30% Bacillus spp., 0-10% Pseudomonas spp., 0-10% Flavobacterium spp., and 0-20% yeast. (c) Following the early stage of pickling, mainly halophilic bacteria such as Bacillus subtilis, Leuconostoc mesenteroides, Pediococcus halophilus and Sarcina litoralis, were found to exhibit an effect on the fermentation of pickle and their enzyme activities were in direct concern in fermentation of pickles. (d) Among the bacteria participating in the fermentation, Sarcina litoralis 8-14 and 8-16 strains were in need of high nutritional requirement and the former was grown only in the presence of purine, pyrimidine and cystine and the latter purine, pyrimidine and glutamic acid. 2. Enzyme characteristics studied in relation to the raw materials and the concerned microbes isolated are as follows: (a) A small amount of protease was found in the raw materials and 30-60% decrease in protease activity was demonstrated at 7% salt concentration. (b) Protease activity of halophilic bacteria, Bacillus subtilis 7-6, 11-1, 3-6 and 9-4 strains, in the complete media decreased by 10-30% at the 7% salt concentration and that of Sarcina litoralis 8-14 and 8-16 strains decreased by 10-20%. (c) Proteins in the raw materials were found to be hydrolyzed to yield free amino acids by protease in the fermenting microbes. (d) No accumulation of flavorous 5'-mononucleotides was demonstrated because RNA-depolymerase in the raw materials and the pickles tended to decompose RNA into nucleoside and phosphoric acid. (e) The enzyme produced in Bacillus subtilis 3-6 strain isolated from the salted clam pickles, was ascertained to be 5'-phosphodiesterase because of its ability to decompose RNA and thus accumulating 5'-mononucleotide. (f) It was demonstrated that the activity of phosphodiesterase in Bacillus subtilis 3-6 strain was enhanced by some components in the corn steep liquor and salted clam pickle. The enzyme activity was found to decrease by 10-30% and 40-60% at the salt concentration of 10% and 20%, respectively. 3. Quantitative data for free amino acids in the pickles are as follows: (a) Amounts of acidic amino acids such as glutamic and aspartic acids in salted clam pickle, were observed to be 2-10 times other pickles and it is considered that the abundance in these amino acids may contribute significantly to the specific flavor of this food. (b) Large amounts of basic amino acids such as arginine and histidine were found to occur in salted yellow tail pickle. (c) It is much interesting that in the salted cuttlefish pickle the contents of sulfur-containing amino acids were exceedingly high compared with those of others: cystine was found to be 17-130 times and methionine, 7-19 times. (d) In the salted oyster pickle a high content of some essential amino acids such as lysine, threonine, isoleucine and leucine, was demonstrated and a specific flavor of the pickle was ascribed to the sweet amino acids. Contents of alanine and glycine in the salted oyster pickle were 4 and 3-14 times as much as those of the others respectively. 4. Analytical data for 5'-mononucleotides in the pickles are as follows: (a) 5'-Adenylic acid and 3'-adenylic acid were found in large amounts in the salted yellow tail pickle and 5'-inosinic acid in lesser amount. (b) 5'-Adenylic acid, especially 3'-adenylic acid predominated in amount in the salted oyster pickle over that in the other pickles. (c) The salted cuttlefish pickle was found to contain only 5'-adenylic acid and 3'-adenylic acid. It has become evident from the above fact that clam and the invertebrate lack of adenylic deaminase and contain high content of adenylic acid. Thus, they were demonstrated to be the AMP-type. (d) 5'-Inosinic acid was contained in the salted yellow tail pickle in a significant concentration, and it might be considered to be IMP-type. 5. Comparative data for flavor with regard to the flavorous amino acids and the contents of 5'-mononucleotides are: (a) A specific flavor of salted yellow tail pickle was ascribed to the abundance in glutamic acid and aspartic acid, and to the existence of a small amount of flavorous 5'-inosinic acid. The combined effect of these components was belived to exhibit a synergistic action in producing a specific fiavor to the pickle. (b) A specific flavor of salted clam pickle has been demonstrated to be attributable to the richness in glutamic acid and aspartic acid rather than to that of 5'-mononucleotides.

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Screening of conjugated linoleic acid (CLA) producing Lactobacillus plantarum and production of CLA on soy-powder milk by these stains (공액리놀레산 생성 Lactobacillus plantarum 선발 및 이를 이용한 콩-분말 두유에서 공액리놀레산 생산)

  • Kim, Baolo;Lee, Byong Won;Hwang, Chung Eun;Lee, Yu-Young;Lee, Choonwo;Kim, Byung Joo;Park, Ji-Yong;Sim, Eun-Yeong;Haque, Md. Azizul;Lee, Dong Hoon;Lee, Jin Hwan;Ahn, Min Ju;Lee, Hee Yul;Ko, Jong Min;Kim, Hyun Tae;Cho, Kye Man
    • Korean Journal of Microbiology
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    • v.51 no.3
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    • pp.231-240
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    • 2015
  • In this study, a total of 16 conjugated linoleic acid (CLA) producing lactic acid bacteria (LAB) were isolated from fermented foods. Among those strains, the S48 and P1201 strains were capable of producing higher CLA contents than other LABs. The two strains were classified as Lactobacillus plantarum based on morphological, physiological, chemotaxonomic, and molecular-genetic properties. The survival rates of these strain appeared to be 59.57% and 62.22% under artificial gastric conditions after 4 h at pH 2.5, respectively. These strains produced the cis-9, trans-11, and trans-10, cis-12 CLA isomers from 8% skim milk medium supplemented with the different free LA concentration at $37^{\circ}C$ for 48 h and the production of two CLA isomers constantly increased in the growth until 48 h of incubation. After 48 h of fermentation, the levels of CLA appeared highest in steamed soy-powder milk than fresh and roasted soy-powder milks. In particular, the CLA contents were produced $183.57{\mu}g/ml$ and $198.72{\mu}g/ml$ from steamed soy-powder milk after fermentation (48 h) with S48 and P1201 strains, respectively.

Anti-Obesity Effects of Fermented Soybean Oils in 3T3-L1 Pre-Adipocytes and High Fat Diet-Fed C57BL/6J Mice (발효콩 유지의 3T3-L1 지방전구세포와 고지방식이를 급여한 C57BL/6J 생쥐에 대한 항비만 효과)

  • Kim, Seon-Woong;Kim, Nam-Seok;Oh, Mi-Jin;Kim, Ha-Rim;Kim, Min-Sun;Lee, Da-Young;Yoon, Suk-Hoo;Jung, Mun-Yhung;Kim, Hun-Jung;Lee, Chang-Hyun;Oh, Chan-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.3
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    • pp.279-288
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    • 2017
  • This study investigated the manufacturing of fermented soybean oil using a fermenting strain commonly processed for soybeans [Bacillus amyloliquefaciens (BA), Bacillus subtilis (BS), Lactobacillus acidophilus (LBA), and B. subtilis+L. acidophilus (BLO)] and evaluated its anti-obesity activities. Cytotoxicity of four kinds of fermented soybean oils was not observed in 3T3-L1 preadipocytes at 10 and $50{\mu}g/mL$. Triglyceride content was reduced by 20.6% in the BLO group at a treatment concentration of $50{\mu}g/mL$. The simultaneous treatment of fermented soybean oil and differentiation induction medium decreased $PPAR{\gamma}$ and $C/EBP{\alpha}$ gene expression at a concentration of $50{\mu}g/mL$ and blocked adipocyte differentiation by increasing adiponectin gene expression. The inhibitory effect of adipocyte differentiation was greatest in the BLO group. C57BL/6J mice were examined for 4 weeks after being separated into seven groups [normal diet group (N), high fat diet group (C), group fed high fat diet combined with regular soybean oil (SO), group fed non-fermented soybean oil (NF), and groups fed high fat diet combined with 5% fermented soybean oil (BA, BS, LBA, and BLO)] to identify the effects of soybean oil on body weight, serum lipid, adiponectin, insulin, and leptin levels in mice with high fat diet-induced obesity. The body weight and serum lipid level of the C group increased drastically compared to those of the N group. In contrast, the group fed a diet combined with fermented soybean oil showed decreases in weight, serum total cholesterol, LDL-cholesterol, and triglyceride levels compared to those of the C group. Moreover, soybean oil was found to be effective in the BLO group. In conclusion, fermented soybean oil has positive effects in prohibiting adipocyte differentiation increased by high fat diet and improving serum lipid composition. Therefore, fermented soybean oil can be used as a functional food material with anti-obesity activity.

Symbiotic effectiveness and intrinsic antibiotic resistance of Rhizobium meliloti populated in Korean pasture soils (국내(國內) 초지토양(草地土壤)에 분포(分布)한 Rhizobium meliloti의 질소고정력(窒素固定力)과 항균제반응(抗菌劑反應) 특성(特性))

  • Kang, Ui-Gum;Ha, Ho-Sung;Jung, Yeun-Tae
    • Applied Biological Chemistry
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    • v.35 no.3
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    • pp.179-185
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    • 1992
  • Rhizobium meliloti populated in five Korean pasture soils were characterized by symbiotic effectiveness and intrinsic antibiotic resistance using whole-soil inoculum and 11 antibiotics, respectively. Most probable number (MPN) of naturalized rhizobia counted with alfalfa Vernal[Medicago sativa (L.)] as a host ranged $1.7{\times}10^2\;cells/g$. soil(Chunghyo, Kyeongiu)-$1.0{\times}10^5\;cells/g$. soil(Gampo, Kyeongiu) and ended to be positively associated with soil pH. On the whole, the effectiveness of population as compared to TAL mix inoculum (TAL 380+TAL 1372+TAL 1373) was very low. Nevertheless, there were two highly effective strains, YCK 539 and YCK 542, which were not inferior to TAL 1372, from Ogpo, Dalseong among the total of 30 of 6 isolates per each soil. As long as mean $N_2$ fixing ability of each soil isolate, the isolates from Hyeongog, Kyeonju were outstanding and the rest were in order of Ogpo, Dalseong>Chunghyo, Kyeongju>Hwaweon, Dalseong>Gampo, Kyeongiu. Isolates as a whole were resistant to erythromycin(67崙), nalidixic acid(77%), and streptomycin sulfate(8051), which had the concentration of $100\;{\mu}g/ml$, $160\;{\mu}g/ml$, and $10\;{\mu}g/ml$, respectively and divided into 14 patterns of resistance. Association between resistances in each soil was not clear. And there was no relationship of resistance pattern to effectiveness. The best effective strain YCKa 542 exclusively fell into No. X pattern having resistance to erythromycin, nalidixic acid, and neomycin sulfate.

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