• Title/Summary/Keyword: bovine milk sample

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Antimicrobial Susceptibility of Staphylococci sp. Isolated from Bovine Milk (우유에서 분리된 포도상구균속 세균의 항생제 감수성)

  • Kim, Ji-Hoon;Ko, Mun-Joo;Kim, Ka-Hee;Lee, Seung-Hoon;Choi, Sung-Sook
    • Korean Journal of Microbiology
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    • v.46 no.4
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    • pp.341-345
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    • 2010
  • The prevalence and antimicrobial susceptibilities of Staphylococcal isolates from bovine milk samples were assessed. From January 2009 to October 2009, a total 287 bovine milk samples were randomly collected from 15 stock raising farms located in northern area of Kyunggi province and cultured for the presence of Staphylococci spp. A total 79 staphylococcal isolates were recovered from the milk samples. The predominant isolates were S. aureus (43.03%) and S. chromogenes (24.05%). Antimicrobial resistance patterns of 79 Staphylococcal isolates against ampicillin, chloramphenicol, ciprofloxacin, erythromycin, gentamicin, oxacillin, teicoplanin, tetracyclin, and vancomycin were tested. Staphylococcal isolates revealed the highest resistance to ampicillin (56.96%) and oxacillin (39.23%). Of 31 oxacillin resistance strains, 8 strains carry mecA gene which is responsible for methicillin resistance.

Identification of Bovine Pregnancy-Specific Whey Proteins using Two-Dimensional Gel Electrophoresis

  • Han, Rong-Xun;Choi, Su-Min;Kim, Myung-Youn;Quan, Yan Shi;Kim, Baek-Chul;Diao, Yun Fei;Koqani, Reza;Park, Chang-Sik;Jin, Dong-Il
    • Reproductive and Developmental Biology
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    • v.32 no.4
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    • pp.255-261
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    • 2008
  • The early diagnosis of bovine pregnancy is an essential component of successful reproductive planning on farms, because lack of bovine pregnancy over the long term results in reproductive failure and low milk yield-the latter of which is a special concern on dairy farms. This study was designed to identify early pregnancy-specific whey proteins in bovine, by comparing milk samples collected from cattle during pregnancy (Days 30 and 50) and from non-pregnant cattle. In this study, differentially expressed proteins in five pregnant and five non-pregnant Holstein dairy cattle were investigated and compared, using proteomics analysis. The first dimension was applied to a pH $3.0{\sim}10.0$ strip, by loading a 2-mg milk protein sample. After the second-dimension separation was performed, the gels were stained with colloidal Coomassie brilliant blue. The stained gels were scanned and the images were analyzed, to detect variations in protein spots between non-pregnant and pregnant cattle milk protein spots, using ImageMaster, this was followed by analysis with MALDI TOF-MS. Analysis of the 2-DE gel image resulted in a total of approximately $500{\sim}600$ protein spots, of which 12 spots were differentially expressed, six spots were up-regulated, and four spots were down-regulated; two spots were identified as pregnancy-specific proteins. These proteins were identified as lactoferrin, NA-DH dehydrogenase subunit 2, albumin, serum albumin precursor and transferrin. Our results via 2-D PAGE analysis revealed composite profiles of several milk proteins related to early bovine pregnancy, implying the possible use of these milk proteins in the early detection of bovine pregnancy.

Changes of Bovine Colostral Immunoglobulin G on Processing Conditions (가공처리조건이 초유 Immunoglobulin G의 변화에 미치는 영향)

  • 이수원;양동훈;황보식;이승환
    • Food Science of Animal Resources
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    • v.21 no.3
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    • pp.265-271
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    • 2001
  • We investigated changes of immunoglobulin G (IgG) concentrations by heating and drying condition. Also it is performed to group for commercial product by promoting of IgG preservation and reducing of protein denaturation. The result was that content of IgG in colostrum was higher than normal milk. Especially, IgG content of colostrum within 12 hrs after parturition was over 44.67mg/ml and it is 60 times of normal milk. IgG contents was reduced rapidly according as passage of the time. IgG content of the sample heating at 30min at 65$^{\circ}C$ was still a little higher that heating for 10sec at 72$^{\circ}C$. IgG denaturation of heat treatment at 100$^{\circ}C$ for 10sec was lower than at 85$^{\circ}C$ for 30min. We investigated the changes of IgG concentrations of kinds of market milk different with heating processing. This result showed that IgG denaturation ratio by ultra high temperature pasteurization (UHT) was higher than long time low temperature pasteurization (LTLT). On the other hands, IgG content by spray drying was 14.5mg/g and freezing drying was 10.8mg/g. It showed that denaturation of protein content by freezing drying was more than spray drying.

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Enzyme-Linked Immuno-Sorbent Assay for Bovine Caseins (우 Casein의 면역효소분석법)

  • 염행철
    • Korean Journal of Animal Reproduction
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    • v.16 no.2
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    • pp.87-102
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    • 1992
  • A rapid, sensitive, and specific enzyme-linked immuno-sorbent assay (ELISA) for bovine casein was developed. Biotinylated casein and peroxidase-conjugated avidin were used in the assay with antibody separated from yolks of immunized hens. Caseins were biotinylated with sulfo-N-hydroxy succinimido biotin and peroxi-dase-conjugated avidin bound the biotinylated casein which became bound to immobilized anti-body on a microplate. The antibodies were specific for bovine $\alpha$- and $\beta$-caseins, and their cross-reactivities with whey proteins, IgG, and serum albumin from bovine were not detectable by ELISA and Western blot. Various sensitivities ranging from 2ng/ml to 20${\mu}\textrm{g}$/ml of casein were achieved, and were controlled by adding vanous concentrations of the biotinylated casein. Parallelism was observed between standard and sample curves. The coefficients of variation of intra-assays and inter-assays from the most sensitive assay were 5.5 and 5.7%, respectively, at the 50% displacement. Casein contents of peripaturient milk samples showed that casein secretion rapidly increased 3d prepartum.

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Distribution of Multidrug Efflux Pump Genes in Enterococci spp. Isolated from Bovine Milk Samples and Their Antibiotic Resistance Patterns (원유 시료에서 분리한 장알균속 세균의 다중약물 유출 펌프(Multidrug Efflux Pump) 유전자의 분포도와 항생제 내성 패턴)

  • Kang, SoWon;Lee, SangJin;Choi, SungSook
    • Korean Journal of Microbiology
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    • v.49 no.2
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    • pp.126-130
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    • 2013
  • The major aim of this study was to investigate the distribution of genes that encode multidrug efflux pumps in Enterococci spp. isolates from bovine milk samples and antibiotic resistance patterns of these strains. Of the 245 isolates, 44.1% showed ampicillin resistance, 79.2% showed erythromycin resistance, 76.3% showed tetracycline resistance and 36.3% showed chloramphenicol resistance. In case of vancomycin and ciprofloxacin, all of the isolates were susceptible to these antibiotics. Of the 245 enterococcal isolates, 82.1% have MFS type eme(A) gene, 72.7% have ABC type efr(A) gene, 77.1% have ABC type efr(B) gene, and 71.8% have ABC type lsa gene. In case of Enterococcus faecalis, the original strain for these genes, 92.5% have eme(A), 87.4% have efr(A), 88.4% have efr(B), and 88.4% have lsa. Interestingly, in case of different species of Enterococci, eme(A) was also detected in four strains of E. faecium, seven strains of E. avium, four strains of E. durans and two strains of E. raffinosus. efr(A) was also detected in two strains of E. faecium and two strains of E. durans and efr(B) was also detected in four strains of E. faecium, five strains of E. avium and four strains of E. durans. This means the possibility of co-transfer of resistance genes between Enterococci species in natural environment. These results are the first report describing the presence of same multidrug efflux pumps in different species of Enterococci in Korea.

An Enzyme-Linked Immunosorbent Assay for $Aflatoxin\;M_1$ in Cow's Milk without a Cleanup Procedure (희석에 의한 우유 중 $Aflatoxin\;M_1$의 효소면역측정법)

  • Shon, Dong-Hwa;Lim, Sun-Hee;Lee, Yin-Won
    • Korean Journal of Food Science and Technology
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    • v.28 no.6
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    • pp.1184-1187
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    • 1996
  • A simple and rapid detection system for $aflatoxin\;M_1\;(AFM_1)$ in cow's milk by an enzyme-linked immunosorbent assay (ELISA) was developed. Specific antibodies against $AFM_1$, conjugated to bovine serum albumin $(AFM_1-BSA)$ were raised in rabbits and purified. The cross-reactivities of the antibodies against aflatoxin analogs were less than 29.9%. When a competitive direct ELISA (cdELISA) for $AFM_1$, established by use of the antibodies was applied to the spike test of $AFM_1$ onto uncontaminated cow's milk, the assay recovery was unstable unless cow's milk was diluted to 40% (2:3) with phosphate buffered saline (PBS). In that condition of sample dilution, the mean ELISA recovery of $AFM_1$, from the cow's milk was 113% (coefficient of variation (CV) of each recovery percentage, 8.2%) in the range of $0.3{\sim}3.0\;ppb$. These results showed that the ELISA system could be a convenient tool to monitor the contamination of AFM1 more than 0.5 ppb in cow's milk (FDA allowance limit) easily.

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Quantitative Analysis of Milk-Derived microRNAs and Microbiota during the Manufacturing and Ripening of Soft Cheese

  • Oh, Sangnam;Park, Mi-Ri;Ryu, Sangdon;Maburutse, Brighton E.;Kim, Ji-Uk;Kim, Younghoon
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1566-1575
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    • 2017
  • MicroRNAs (miRNAs) are abundant in bovine milk and milk derived from other livestock, and they have functional roles in infants and in the secretion process of mammary glands. However, few studies have evaluated miRNAs in dairy processes, such as during cheese making and ripening. Thus, we investigated the characteristics of milk-derived miRNAs during the manufacturing and ripening of Camembert cheese as well as the microbiota present using the quantitative reverse transcription polymer chain reaction (RT-qPCR) and 16S rRNA pyrosequencing, respectively. Pyrosequencing showed that the cheese microbiota changed dramatically during cheese processing, including during the pasteurization, starter culture, and ripening stages. Our results indicated that the RNA contents per $200mg/200{\mu}l$ of the sample increased significantly during cheese-making and ripening. The inner cheese fractions had higher RNA contents than the surfaces after 12 and 22 days of ripening in a time-dependent manner (21.9 and 13.2 times higher in the inner and surface fractions than raw milk, respectively). We performed a comparative analysis of the miRNAs in each fraction by RT-qPCR. Large amounts of miRNAs (miR-93, miR-106a, miR-130, miR-155, miR-181a, and miR-223) correlated with immune responses and mammary glands were present in aged cheese, with the exception of miR-223, which was not present on the surface. Considerable amounts of miRNAs were also detected in whey, which is usually disposed of during the cheese-making process. Unexpectedly, there were no significant correlations between immune-related miRNAs and the microbial populations during cheese processing. Taken together, these results show that various functional miRNAs are present in cheese during its manufacture and that they are dramatically increased in amount in ripened Camembert cheese, with differences according to depth.

A study on simultaneous determination of residual sulfonamides in livestock productions by high performance liquid chromatography (HPLC를 이용한 축산식품중 잔류 설폰아미드제의 동시분석법 연구)

  • 황래홍;김영수;윤은선;김기근;이규학
    • Korean Journal of Veterinary Service
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    • v.18 no.3
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    • pp.13-28
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    • 1995
  • This study was carried out to explore the most sensitive and useful method for simultaneous determination of five sulfa drugs(sulfamethazine, sulfamerazine, sulfamonomethoxine, sulfadimethoxine, sulfaquinoxaline) in livestock productions(pork muscle, bovine muscle, chicken muscle, milk ) by HPLC with UV detector and reverse phase column. The results obtained were as follows:1. For mobile phase acetonitrile-0.01M ammonium acetate (23:77) showed more applicable sensitivity and retention times than acetonitrile-1% acetic acid(23:77). Thus acetonitrile-0.01M ammonium acetate(23:77) selected and applied to the modification test, from which it was found pH 6.75 was the most adequate. 2. Optimal wavelength of UV for SMT(sulfamethazine), SMR(sulfamerazine), SMM(sulfamonomethoxine), SD(sulfadimethoxine), and SQ(sulfaquinoxaline) were 266, 266, 265, 269 and 250nm, respectively, and that for simultaneous application it was 263nm. 3. The average recovery rate by extractant(chloroform, dichloromethane, chlorform+dich-loromethane) in the classic method was not significantly different(p>0.05) but that by chloroform higher than the others. Thus chloroform was found to be adequate as extractant in this classic method. 4. The average recovery rate was 86.5% by the MSPD(matrix solid phase disperse) method, which was significantly higher than that by the classic method(p<0.05). Also the recovery rates by method were significantly different(p<0.05) in accordance with sample and type of drug. The MSPD method was much superior to classic method on clean-up.

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Prevalence and risk factors of subclinical bovine mastitis in some dairy farms of Sylhet district of Bangladesh

  • Kahir, Md. Abdul;Islam, Md. Mazharul;Rahman, A.K.M. Anisur;Nahar, A.;Rahman, Md. Siddiqur;Son, Hee-Jong
    • Korean Journal of Veterinary Service
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    • v.31 no.4
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    • pp.497-504
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    • 2008
  • A cross-sectional study was undertaken to report prevalence and to identify risk factors of subclinical mastitis of dairy cattle in Sylhet district of Bangladesh. Among 325 dairy farms of the district 12 farms(3.7%) were selected conveniently for this study. All the dairy cows of the 12 farms were selected for sample collection. Fresh milk samples from each of the selected dairy cows were collected aseptically in separate sterilized test tube as RF, RH, LF and LH quarter of the udder. Rapid modified White Side Test(WST) was used to detect subclinical mastitis(SCM). Results of WST and data derived from filled in questionnaire were entered in Microsoft Excel 2003 and transferred to $STATA^{(R)}$, version 8.0/Intercooled(Stata Corporation, Texas, USA, 2003). The overall prevalence of SCM and its distribution in different categories of variables in cow and their exact binomial 95% confidence intervals were calculated in $STATA^{(R)}$. Simple bivariable associations among independent variables were investigated by $x^2$ test in $STATA^{(R)}$. Multiple logistic regression analysis with backward elimination method was used to identify risk factors of SCM. To identify significant variation in quarter SCM, linear regression analysis was performed after arcsine transformation of the data. The overall prevalence of SCM found in this study is 54%. Dairy cows with teat lesions had significantly increased SCM(OR=12342, P value=0.000, 95% CI=762, 199798) than others without teat lesions. The Holstein Friesian X Jersey X Sahiwal breed has significantly decreased(OR=0.18, p=0.03, 95% CI 0.04, 0.85) SCM than other breeds. The prevalence of SCM found in this study is in agreement with others. The injury in the teat increases the probability of getting infected with microbes and thereby mastitis. If the prevalence of teat lesion can be decreased the probability of subclinical mastitis will also be decreased. The negatively associated Holstein Friesian X Jersey X Sahiwall breed may help in planning mastitis control program if this finding can be validated by a more powerful case-control or cohort study design.