GC-MS/MS using liquid-liquid extraction (LLE) and C18 cartridges was used to identify and quantify levels of chlorpyrifos, chlorpyrifos-methyl, cypermethrin, deltamethrin and permethrin in bulk raw milk. A calibration curve spanning 10 ng/mL to 200 ng/mL was obtained with a satisfactory correlation coefficient of 0.99. The limits of detection (LOD) and limits of quantitation (LOQ) for chlorpyrifos, chlorpyrifos-methyl, cypermethrin, deltamethrin, and permethrin in the matrix ranged from 0.06 to 1.81 ng/mL and 0.19 to 6.04 ng/mL, respectively. The recoveries of 5 pesticides from spiked samples at 37.5-125 ng/mL ranged from 86.1 to 102.1%. The measurement of uncertainty of the GC-MS/MS method for these five pesticides was developed based on the analytical process and quantification. An analysis method that is easier and faster than the method specified in the Korean food standards codes for analyzing these five pesticides in raw material milk was developed. Moreover, the analytical method for chlorpyrifos, chlorpyrifos-methyl, cypermethrin, deltamethrin, and permethrin in bulk raw milk by GC-MS/MS was established.
The goal of this research was to develop a portable system that could be used to evaluate the quality of milk in real time at a raw milk production site. A real-time portable quality evaluation system for raw milk was developed to enable non-destructive quality evaluation of somatic cell count (SCC), fat, protein, lactose, and total solid (TS) in milk samples. A prediction model of SCC, fat, protein, lactose, and TS was constructed using partial least squares (PLS) and 200 milk samples were used to evaluate the prediction performance of the portable quality evaluation system and high performance spectroscopy. Through prediction model development and verification, it was found that the accuracy of high performance spectroscopy was 90% for SSC, 96% for fat, 96% for protein, 91% for lactose, and 97% for TS. In comparison, the accuracy of the portable quality evaluation system was relatively low, at 90% for SSC, 95% for fat, 92% for protein, 89% for lactose, 92% for TS. However, the measurement time for high performance spectroscopy was 10 minutes for 1 sample, while for the portable quality evaluation system it was 6 minutes. This means that the high performance spectroscopy system can measure 48 samples per day (8 hours), while the portable quality evaluation system can measure 80 (8 hours). Therefore, it was found that the portable qual ity evaluation system enables quick on-site quality evaluation of milk samples.
The current standard solutions for somatic cells used for calibration of electronic somatic cell counts as reference material in raw milk are preserved with bronopol, boric acid, sodium azide, or potassium dichromate, and have a shelf-life of only up to 6 days at 4 ± 2℃. In the present study, a set of somatic cell standard solutions (SCSS) with a stability of 5 months for calibration of electronic instruments was developed. Somatic cells collected from cow’s milk and stored in a bulk tank at a dairy plant were treated with 10% formaldehyde in order to improve stability, and then separated by centrifugation. The resulting somatic cell suspension was preserved with glycerin, thimerosal, and dimethyl sulfoxide, and diluted in 3% processed skim milk solution ranging from 200,000~250,000 (low level), 350,000~ 450,000 (medium level), and 550,000~650,000 (high level) cells/㎖. Each SCSS was verified by direct microscope somatic cell counting (DMSCC), C-reader, and commercial standard samples. The average somatic cell count determined by DMSCC was 248, 214, 226 × 103 cells/㎖, 436, 382, 420 × 103 cells/㎖, and 612, 595, 609 × 103 cells/㎖. The coefficient of variation representing the repeatability of DMSCC decreased as the number of cells increased, and was <10.0% in almost all SCSS samples (range 4.6~7.1%). No statistically significant difference in somatic cell concentration was observed after storage at refrigeration temperature (2~6℃) over a period of 22 weeks (5 months). The stabilized SCSS may be useful as a reference material for determination of somatic cell count and quality control in testing of bovine raw milk.
Antibiotic resistance in animal isolates of enterococci is a public health concern, because of the risk of transmission of antibiotic-resistant strains or resistance genes to humans through the food chain. This study investigated phenotypic and genotypic resistances profile of tetracycline in 245 Enterococcus isolates from bovine milk. A total of 245 enterococci were isolated from 950 milk samples. The predominant strain was E. faecalis (n = 199, 81.2%) and E. faecium (n = 25, 10.2%). E. avium (n = 7, 2.9%), E. durans (n = 6, 2.5%), E. gallinarum (n = 4, 1.6%), and E. raffinosus (n = 4, 1.6%) were also isolated. Of the 245 enterococcal isolates 76.3% (n = 187) displayed tetracycline resistance (≥ 16 μg/ml). Of the 187 tetracycline-resistant isolates, 83.4% (n = 156), 16.1% (n = 30), and 26.7% (n = 50) possessed the genes tet(M), tet(L), tet(S) respectively. While 3.2% (n = 6) of the tetracycline- resistant isolates possessed all three genes tet(M) + tet(L) + tet(S), 8.6% (n = 16), 16.0% (n = 30), and 2.7% (n = 5) of them possessed two genes tet(M) + tet(L), tet(M) + tet(S), and tet(L) + tet(S) respectively. The tetracycline resistance pattern investigated in this study was attributable mainly to the presence of tet(M).
농산물의 안전성 확보와 품질평가를 위해서 신속하고 경제적인 비파괴 검사법에 대한 연구 및 기술개발이 활발하게 이루어지고 있다. 그러나 분광된 빛을 이용하여 내부 품질평가 과정에서 발생할 수 있는 분석 대상체의 성분 변화에 미치는 영향에 대한 연구는 미비한 실정이다. 따라서 본 연구는 분광분석을 이용하여 원유의 성분분석 과정에서 분광된 빛이 원유의 성분 및 체세포에 미치는 영향을 구명하고자 수행되었다. 본 연구에 사용된 원유는 일본 시가현 소재 낙농가에서 채취된 것으로 유지방, 유단백질, 유당, 무지고형분, 총고형분, 유요소, 구연산 및 체세포수를 화학적 방법에 의해 측정하였다. 또한 인위적으로 분광된 빛은 5가지 영역대로 구분하여 원유에 각각 1분, 5분, 10분간 주사하였다. 연구결과, 400 nm이하 파장대인 자외선 영역에서 유지방이 2.6% 증가되는 경향이 있었고, 체세포수도 9.0% 증가된 것으로 분석되었다. 그러나 다른 원유 성분에는 변화가 없는 것으로 나타났다. 따라서 자외선 영역의 빛을 원유 품질 평가에 적용하기 위해서는 보다 다양한 원유 시료에 대한 추가 검증이 필요할 것으로 사료된다.
From April 2008 to January 2009, a total 458 raw milk samples were randomly collected from 15 stock raising farms located in northern area of Kyunggi province and cultured for the presence of Enterococci spp. A total 170 enterococcal isolates were recovered from the raw milk samples. Enterococcus faecalis was predominant species recovered (64.7%), followed by E. faecium (18.8%), E. avium (5.9%), E. gallinarum (5.9%) and E. durans (4.7%). Antimicrobial resistance patterns of 170 Enterococci spp. against ampicillin, erythromycin, tetracyclin, chloramphenicol, vancomycin, ciprofloxacin and streptomycin were tested. According to the result, they showed high level resistance to erythromycin and streptomycin (82,9% and 93,5% respectively), moderately resistance to ampicillin, chloramphenicol and tetracyclin (50%, 45.9% and 32%, respectively) but fortunately, vancomycin and ciprofloxacin are still effective against this species.
본 연구의 목적은 L. monooytogenes에 대한 LP system의 항균효과를 측정하기 위하여 수행되었다 초기 접종수준(10², 10⁴, 10^(7) CFU/ml), LP의 농도는(10, 20, 30 ppm), 그리고 저장온도(5, 10, 15℃), 배지종류(TSB-YE, UHT milk)에 따라 L. monocytogenes에 대한 항균효과를 측정 비교한 결과 초기접종수준을 10²cfu/ml로 하였을 때와 LP농도는 10 ppm 및 5℃로 배양에서 항균력이 높게 나타났다. UHT milk를 이용한 LP system의 항균효과는 Tryptic soy agar를 이용한 시험결과와 비슷하게 나타났다.
본 연구는 지방산 함량 및 배합사료와 조사료 비율을 기초로 한 유기농 사료 급여가 홀 스타인 착유우에서 생산된 원유의 CLA 및 지방산 함량에 미치는 연구를 조사하기 위한 목 적으로 실시하였다. 총 290두의 홀스타인 착유우를 산차 및 유량에 따라 3개 group으로 나 누었다. 대조구는 C16:00, C18:2 그리고 SFA를 높게 설계하였고, 처리구 1은 C18:1, C18:2 그리고 UFA 함량을 높게 설계하였으며 처리구 2는 MUFA, C18:3 그리고 PUFA 함량을 높 게 설계하였다. 결과를 요약하면 다음과 같다. 유기농 원유 내 C16:0 함량은 처리구 2에서 가장 높은 것으로 나타났다(p<0.05). 그 이유는 반추위 내 미생물의 de novo 생합성 때문인 것으로 판단된다. 처리구 2의 C18:0 함량은 7.92%로 대조구(11.39%)와 처리구 1(10.88%)보 다 높았다(p<0.05). CLA 함량도 처리구 2가 처리구 1이나 대조구에 비하여 높은 것으로 나 타났다(p<0.05). 원유 내에서 검출된 대부분의 CLA는 착유우 유선조직내의 △9-desaturase에 의하여 합성된 것으로 판단된다. n-3/n-6 비율도 처리구 2에서 가장 높은 것으로 나타났다 (p<0.05). 본 연구 결과를 종합해 보면, 착유우에게 혼합 목건초 등을 급여하면 CLA, n-3 농 도는 증가하며 C18:0 농도는 낮아지는 것으로 조사되었다. 본 연구는 유기농 인증된 조사 료 및 농후사료를 이용하여 결과를 도출하였다. 그러나 유기농 사료를 사용할 경우에만 원 유 내 CLA 및 n-3 농도는 증가한다고 볼 수 없다. 원유 내 고농도의 CLA 및 n-3 지방산 생 산을 위해서는 반추위 미생물 및 유선세포의 지방 대사를 통한 CLA 생산 메커니즘에 대한 충분한 이해와 급여 사료 내 지방산 구성 등이 중요한 것으로 판단된다.