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Characteristics of Histamine Forming Bacteria from Tuna Fish Waste in Korea (국내 참치 부산물 내 히스타민 생성 주요 세균의 특성 구명)

  • Bang, Min-Woo;Chung, Chang-Dae;Kim, Seon-Ho;Chang, Moon-Baek;Lee, Sung-Sil;Lee, Sang-Suk
    • Journal of Life Science
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    • v.19 no.2
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    • pp.277-283
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    • 2009
  • Biogenic amines are generally formed through the decarboxylation of specific free amino acids by exogenous decarboxylases released by microbial species associated with the fish products and fermented feeds. This study was conducted to investigate the properties of e tuna waste regarding the control of degradation of biogenic amines (histamine, tyramine, tryptamine, putrescine, and cadaverine) that might be related with the anti-nutritional factor of the tuna waste that is used for manufacturing domestic fish meal. The values of pH and the salt content were 6.51, 3.35% in tuna waste and 5.58 and 5.83% in tuna fish meal, respectively. The strains and dominant bacteria tested in the tuna waste sample were 9.20, 9.29, 5.67, 7.82 and 7.58 log CFU/g of total bacteria, aerobic plate count (APC), total coliform (TC), Lactobacillus spp. and Bacillus spp., respectively. The main histamine forming-bacteria (HFB) in tuna waste were detected by silica gel thin-layer chromatography (TLC) and 7 histamine-forming bacterial species were isolated among microbes grown in selective medium. The histamine concentration was determined by detection of fluorescence of ο-phthaldialdehyde (OPA) derivatives using HPLC and the date were used to reconfirm the identities of the amine-producing bacteria. The 15 histamine- forming bacteria strains grown in trypicase soy broth (TSB) supplemented with 1% L-histidine (TSBH) were identified as Lactococcus(L.) lactis subsp. lactis, Klebsiella pneummonlae, L. garvieae 36, Vibrio olivaceus, Hafnia alvei and L. garvieae which were main dominant amine - producing strains, and Morganella morganii identified by 16S ribosomal RNA (rRNA) sequencing with PCR amplification. A Phylogenetic tree generated from the 16S rRNA sequencing data showed different phyletic lines that could be readily classified as biogenic amine forming gram-positive and negative bacteria.

Systems for Production of Calves from Hanwoo(Korean Native Cattle) IVM/IVF/IVC Blastocyst I. Hanwoo IVM/IVF /IVC Blastocyst Cryopreserved by Vitrification (체외생산된 한우 배반포기배로부터 송아지 생산을 위한 체계 I. 체외생산된 한우 배반포기배의 초자화 동결보존)

  • Park, S. P.;Kim, E. Y.;Kim, D. I.;Park, N. H.;Y. S. Won;S. H. Yoon;K. S. Chung;J. H. Lim
    • Korean Journal of Animal Reproduction
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    • v.22 no.4
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    • pp.349-357
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    • 1998
  • This study was to investigate whether the viability of Hanwoo IVM/IVF/IVC blastocysts was maintained after vitrification and thawing. In vitro produced Hanwoo blastocysts were vitrified by two-step method: equilibrated in EG20 for 3 min, and then exposed in EFS40 [40% ethylene glycol (EG), 18% ficoll and 10.26% sucrose in mDPBS containing 10% FBS ]and vitrified in L$N_2$for 30 - 45 sec. After thawing, in vitro survival was assessed as the re-expanded and hatched rates at 24 hand 48 h, respectively. The results obtained in these experiments were summarized as follows: From the 12 replicates, 52.5% of Hanwoo blastocysts were produced in vitro at day 7 after IVF. When the effects of freezing solution to the embryo survival were examined, there is no significant toxicity in exposure (100.0, 73.8%) compared to that af control group (100.0, 87.0%). However, when embryos were vitrified, high survival (86.2, 55.4%) was obtained although it was significantly lower than those of exposure and control group (p<0.05). When the in vitro survival of vitrified embryos according to developmental stage and culture day were examined, it showed that more advanced embryo stage exhibited a significantly higher survival rate irrespective of culture day (p<0.05). Also, even in the same development stage, the in vitro survival of day 7 embryos (re-expanded: 75.0~87.5%, hatched: 21.4~66.7%) was higher than those of day 8 embryos(re-expanded: 58.6~78.3%, hatched: 10.3~52.2%). Therefore, these results suggested that in vitro produced Hanwoo blastocysts can be successfully cryopreserved by simple two-step vitrification method using EFS40 freezing solution, particularly at the expanded and early hatching blastocyst stage regardless of embryo culture duration (day 7 or day 8 after IVF).

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Systems for Production of Calves from Hanwoo(Korean Cattle) IVM/IVF/IVC Blastocyst II. Simple, Efficient and Successful Vitrification of Hanwoo Blastocyst (체외생산된 한우 배반포기배로부터 송아지 생산을 위한 체계 II. 한우 배반포기배를 간편하면서 효율적이고 성공적으로 유리화 동결하는 방법)

  • Kim, E.Y.;Kim, D.I.;Park, N.H.;Weon, Y.S.;Nam, H.K.;Lee, K.S.;Park, S.Y.;Yoon, S.H.;Park, S.P.;Chung, K.S.;Lim, J.H.
    • Korean Journal of Animal Reproduction
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    • v.23 no.4
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    • pp.281-291
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    • 1999
  • The objective of this study was to optimize the freezing/thawing method of in vitro produced Hanwoo blastocysts. Day 7 blastocysts after IVF were vitrified using EFS40 (40% ethylene glycol, 18% ficoll, 0.3 M sucrose and 10% FBS added m-DPBS) as a freezing solution and electron microscope (EM) grid (V-G) or straw (V-S) as an embryo container. In both method, freezing/thawing were treated by 2-step, treatment time was required in V-G method and V-S method, for 2 min / 3 min and 3.5 min / 10 min, respectively. Embryo survival was assessed as re-expanded and hatched rates at 24 h and 48 h after warming, respectively. The results obtained in these experiments were summarized as follows: when the effect of exposure in vitrification solution and chilling injury from freezing procedure on in vitro produced expanded blastocysts were examined, at 24 h after warming, embryo survival in exposure group (100.0%) was not different compared to that in control group (100.0%), although those results were significantly different with two vitrified groups (V-G: 87.8, V-S: 77.8%) (P<0.001). However, at 48 h after warming, hatched rates of V-G group (67.8%) were significantly higher than those of V-S group (53.3%) (P<0.05). In addition, this hatched rate in V-G group was not different with that in exposure group (73.3%). When the effects of embryo developmental stage (early, expanded and early hatching blastocysts) and embryo container (EM grid and straw) to the in vitro survival of vitrified-warmed day 7 Hanwoo blastocysts were simultaneously examined, fast developed embryos were indicated the better resistance to freezing than delayed developed one, irrespective of embryo containers (early; 57.1 & 24.4%, expanded; 84.7 & 60.6%, early hatching; 91.7 & 80.0%) (P<0.001). Especially, in expanded and early hatching blastocysts, embryo survival of V-G group (67.8, 95.0%) was significantly higher than those of V-S group (53.0, 65.0%) at 48 h post warming, respectively (P<0.05, P<0.001). Therefore, this study indicates that Hanwoo blastocysts can be cryopreserved more simple, efficient and successful by vitrification method using EM grid.

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Sequence and Genetic Variation of Mitochondrial DNA D-loop Region in Korean Cattle (한우 Mitochondrial DNA D-loop 영역의 염기서열 및 유전변이)

  • Chung, E.R.;Kim, W.T.;Kim, Y.S.;Lee, J.K.;Han, S.K.
    • Journal of Animal Science and Technology
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    • v.44 no.2
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    • pp.181-190
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    • 2002
  • This study was performed to determine sequences of the mt DNA D-loop region, including $tRNA^{Pro}$ and $tRNA^{Pre}$ and to analysis sequence variation polymorphism in Korean cattle. The resulting sequencies were compared with previously published sequences for other cattle breeds(GenBank J01394). The PCR was used to amplify an 1142bp between nucleotides 15061 and 404 within the D-loop region of mt DNA using specific primers. Korean cattle showed 24 polymorphic sites by nucleotide substitutions and insertions of single base pairs. About 50% of polymorphic sites were found in positions 16042 to 16122 with the most variable region. Among these polymorphic sites, variations at 16055, 16230 and 16260 bp were detected as new sequence variants in Korean cattle. These specific polymorphic sites have not been reported in the Japanese black cattle and European cattle. Therefore, mt DNA variants in the D-loop region may be used as genetic markers for specifying Korean cattle. The frequencies of positions 169, 16302, 16093, 16042, 16119 with a high level of sequence polymorphism were 0.81, 0.56, 0.56, 0.50 and 0.43, respectively. In comparison of genetic distances, Korean cattle showed the more closely to European cattle as Bos taurus than Bos indicus such as African and India breeds. In conclusion, these mt DNA sequence polymorphisms in the D-loop region for Korean cattle may be useful for the analysis of cytoplasmic genetic variation and associations with economic important traits and genetic analysis of maternal lineage.

Development of Microsatellite Markers using BAC clone Sequencing on Porcine Chromosome 6q28 - 6q32 (돼지 6번 염색체(6q28 - 6q32)의 BAC clone 염기서열 분석에 의한 Microsatellite Markers 개발)

  • Chang, K.W.;Lee, K.T.;Park, E.W.;Choi, B.H.;Kim, T.H.;Cheong, I.C.;Oh, S.J.
    • Journal of Animal Science and Technology
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    • v.46 no.3
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    • pp.301-306
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    • 2004
  • This study was conducted to develop new markers at the region that was related to QTL affecting intramuscular fat and backfat thickness on chromosome 6q28 - 6q32 in pigs. Dozens of repeated sequences were founded using shotgun sequencing of several BAC clones corresponding to that region, of which five new microstellite markers that identified polymorphism were discovered. The mean number of alleles at each locus observed 2.13(KP0290F2), 4.63(KP0248Cll), 7.38(KP1231C91), 2.75(KPI23IC92) and 6.2S(KP1231C93) in 8 breeds(Landrace, Korean native pig, Duroc, Yorkshire, Berkshire, Wuzhishan pig, Xiang pig, Min pig). The average estimated heterozygosity values at each locus varied from 0.2100(KP0290F2) to 0.8304(KPI23IC91) in all populations. In other hand, the average allele of all loci WlL'I within range of 0.4517(Berkshire) and 0.6957 (Yorkshire). Of these markers, KP0248C11, KP1231C91 and KP1231C93 were identified to have optimal number of alleles, high heterozygosity values and low standard deviation values. Especially, KPI23IC91 and KPI231C93 might be considered as a useful marker for genetic mapping and diversity study.

Nutritional and Microbiol Quality Assessment of Chungmukimbab (충무김밥의 영양적 및 미생물적 품질 평가)

  • Choi Jong-Duck;Hwang Young-Suk;Jeoung In-Geon;Lee Da-Jeong;Park Phil-Sook;Park Mi-Yeon
    • Journal of Food Hygiene and Safety
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    • v.21 no.3
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    • pp.189-195
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    • 2006
  • This research was designed to evaluate the nutritional an microbiol quality assessment of Chungmukimbab purchased from market in Tongyeoung area. Contents of calories, calcium, iron, thiamin and riboflavin in ordinary kimbab and Chungmukimbab were lower than the recommended levels of Korean adult men. So, We suggested that a fruit, beverage and ffod stuff were supplemented to maintain nutritional balance. Total aerobic bacteria and coliform group of just prepared ordinary kimbab and Chungmukimbab samples from market were not significantly different, showing approximately $5.50{\pm}0.38 log_{10} CFU/g,\;2.10{\pm}0.47log_{10}MPN/100g$ in ordinary kimbab, $5.61{\pm}0.42log_{10}CFU/g,\;1.75{\pm}0.34 log_{10} MPN/100g$ in Chungmukimbab, respectively. Total aerobic bacteria of law ingredients of chungmukimbab sample were 3 to $4 log_{10}CFU/g$ in kimbab, seasoning squid and radish roots kimchi, 4 to $5 log_{10}CFU/g$ in boiled fish paste. The coliform groups were 1 to $2 log_{10}$ MPN/100 g in kimbab, seasoning squid and radish roots kimchi, 2 to $3 log_{10}$ MPN/100g in boiled fish paste. Detection rate of E. coli and Staphylococcus aureus counts were 10.0, 12.5% in Chungmu-kimbab, 15.0, 10.0% in seasoning squid, 0, 10.0% in radish roots kimchi respectively, not detected in boiled fish paste samples. During storage at $15^{\circ}C$ for 24 hours, total aerobic counts and coliform groups in ordinary kimbab and Chungmukimbab were increased by the 1.94, $0.97log_{10}CFU/g$, 0.60, 0.72 log10 MPN/100g respectively. Total aerobic counts of Chungmukimbab ingredients increased $0.83{\sim}l.33 log_{10}CFU/g$ at different time

Hazard Analysis for the Application of Good Agricultural Practices(GAP) on Paprika During Cultivation (파프리카의 농산물우수관리제도(GAP)적용을 위한 재배단계의 위해요소 분석)

  • Nam, Min-Ji;Chung, Do-Yeong;Shim, Won-Bo;Chung, Duck-Hwa
    • Journal of Food Hygiene and Safety
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    • v.26 no.3
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    • pp.273-282
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    • 2011
  • This study established hazards which may cause risk to human at farm during cultivation stage of paprika. Samples of plants (paprika, leaf, stem), cultivation environments (water, soil), personal hygiene (hand, glove, clothes), work utensils (carpet, basket, box) and airborne bacteria were collected from three paprika farms (A, B, C) located in Western Gyeongnam, Korea. The collected samples were assessed for biological (sanitary indications and major foodborne pathogens), chemical (heavy metals, pesticide residues) and physical hazards. In biological hazards, total bacteria and coliform were detected at the levels of 1.9~6.6 and 0.0~4.610g CFU/g, leaf, mL, hand or 100 $cm^2$, while Escherichia coli was not detected in all samples. In major pathogens, only Bacillus cereus were detected at levels of ${\leq}$ 1.5 log CFU/g, mL, hand or 100 $cm^2$, while Staphylococuus aureus, Listeria monocytogenes, E. coli O157 and Salmonella spp. were not detected in all samples. Heavy metal and pesticide residue as chemical hazards were detected at levels below the regulation limit, physical hazard factors, such as insects, pieces of metal and glasses, were also found in paprika farms. Proper management is needed to prevent biological hazards due to cross-contamination while physical and chemical hazards were appropriate GAP criteria.

Pathogenic Staphylococcus epidermidis isolated from cultured fingerling of sea bass, Lateolabrax japonicus, in Korea (남해안 양식산 농어, Lateoabrax japonicus 치어에서 분리한 병원성 Staphylococcus epidermidis에 관한 연구)

  • Cha, Yong-Baeg;Yang, Han-Choon;Choi, Sang-Duk;Cho, Jae-Kwon
    • Journal of fish pathology
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    • v.10 no.1
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    • pp.1-14
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    • 1997
  • Major object of this study was to investigate the causative organism of th e diseased cultured fingerling of sea bass, L japonicus. The experimental results are summarized as follows ; Staphylococcus epidermidis, isolated from the liver, kidney, spleen and brain, was considered to be the causative organism. External symptoms of this disease were congestion and hemorrhages in eyes. Anatomical symptoms were hemorrhage of brain, congestion of liver, and slight swelling of kidney and spleen. Growth of the isolates was good on BHIA, HIA and Staphylococcus No. 110. The growth occurred at a range(optimum) of $10\sim45^{\circ}C(35\sim40^{\circ}C)$, 0~9% (1~3%) of NaCl concentration and pH 4~10(8). DNase and coagulase production of all isolated strains were nagative, but was positive in hemolysis. Urease was positive reaction, and novobiocin resistance was nagative. Acid was produced anaerobically from glucose and maltose. Acid was produced aerobically from glucose, galactose, sucrose, maltose and dextrine. But gas was not produced from any carbohydrates. When the isolated strain was injected intramuscularly on fingerling of sea bass, L japonicus, it had virulence at $1.7{\times}10^{10}$ viable cells/$m\ell$ for all fish examined but no virulence at $1.7{\times}10^4$ viable cells/$m\ell$. Bacitracin, Erythromycin and Nofloxacin were observed as bacteriostatic agents to the strain, but Colistin, Gentamicin and Nalidixic acid were not. There were remarkable congestion of the brain, regressive necrosis of the liver, and showed necrosis of the epithelial cells of renal tubules in kidney tissues.

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A study on the noodle quality made from pea starch-wheat composite flour (완두 전분을 첨가한 국수의 품질특성)

  • 김은주;윤재영;김희섭
    • Korean journal of food and cookery science
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    • v.18 no.6
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    • pp.692-697
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    • 2002
  • The purpose of this study is to investigate the characteristics of the doughs and noodles cooked with the pea starch-wheat composite flour which was substituted with 20% and 30% of pea starch for the flour by the mechanical and sensory evaluation. Wheat dough had the most cohesive property among various composite non.(p<0.05) There was no significant differences in weight gain after cooking among various noodles. The more the pea starch was subsituted, the lighter the color was shown by increasing L value. It was also noted that the b value was decreased significantly. While pea starch noodle were more transparent in appearance and less smooth in the texture, corn starch-wheat composite flour noodle was sorter in the texture significantly. There was no significant difference on the hardness between wheat and pea stach composite flour noodles. There were also no significant differences in stickiness, chewiness and overall acceptability among various noodles. Considering mechanical and sensory results, the composite flour with 20% substitution of pea starch for flour was more suitable for the production of the noodle than those of 30% substitution of pea starch.

Soil Fertility Evaluation with Adoption of Soil Map Database for Tobacco Fields (토양도 자료를 활용한 연초 경작지의 비옥도 평가)

  • Hong, Soon-Dal;Park, Hyo-Taek
    • Korean Journal of Soil Science and Fertilizer
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    • v.32 no.2
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    • pp.95-108
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    • 1999
  • Field experiments were conducted in the 101 tobacco fields(51 fields in 1985 and 50 fields in 1986) of chief tobacco producing counties of Chungbuk province(Jincheon, Eumseong, Goesan, and Joongweon counties), Chungnam province(Cheonweon county), and Kyongbuk province (Cheongdo, Seongju, and Andong counties) for two years from 1985 to 1986 in order to evaluate soil fertility using chemical properties and soil map database. Pot experiments also on the same soils were conducted and the results were compared to those of field experiments. The yield of tobacco in the plots of no fertilization was considered as a basic factor representing the soil fertility and was evaluated by nineteen independent variables, that was 9 chemical properties and 10 soil map databases. These independent variables were classified into two groups, 11 quantitative indexes and 9 qualitative indexes, and were analyzed by multiple linear regression(MLR) of SAS by REG and GLM models. The yield of tobacco in the plot of no fertilization showed high variations, e.g. the difference between minimum and maximum yields was about 5.0-5.5 times in the pot experiment and 8.2-14.9 times in the field experiment. The indexes indicating close link between yield of tobacco and soil chemical indexes, was selected but it was not well matched by the years or between pot and field experiments. Also, the standardized partial regression coefficients of quantitative indexes for the yield of field were less than 1.0, suggesting that it is difficult to develop an available single index for the evaluation of soil fertility. Evaluation for the soil fertility of field by MLR was better than that of single regression and it was gradually improved by adding chemical properties, quantitative indexes, and qualitative indexes of soil map. For example, the coefficient of determination ($R^2$) of MLR for the yield of 1985 was increased to 0.422 with chemical indexes, 0.503 by addition of quantitative indexes, and 0.633 by the additional adding of qualitative indexes of soil map, compared to 0.244 of single index, $NO_3-N$ content of soil. Consequently, it is assumed that this approach by MLR with quantitative and qualitative indexes including chemical properties and soil map databases was available as an evaluation model of soil fertility for tobacco field.

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