牛奶中抗生素残留检测试剂盒的研制及应用

对嗜热脂肪芽孢杆菌芽孢的生成、营养基质、各成分添加比例等条件进行了研究,研制出检测牛奶中抗生素残留的96孔板试剂盒。此试剂盒对牛奶中青霉素G的检测极限为3ng/mL,阿莫西林的检测极限为4ng/mL,头孢匹林的检测极限为8ng/mL。同一批奶样检测结果证明,此试剂盒与西班牙ZEU-INMUNOTEC公司同类试剂盒产品质量相当,灵敏度高于国标TTC法,适于推广应用。      

嗜热脂肪芽孢杆菌检测牛乳中抗生素残留

采用嗜热脂肪芽孢杆菌作为指示菌,制备牛乳抗生素检测微孔板,对牛乳中抗生素残留进行检测。结果表明,研制出的抗生素检测微孔板稳定性良好,操作简便、快捷、灵敏度高,结果容易判断。具有广谱性,可检测β-内酰胺类、磺胺类、四环素类和氨基糖苷类等多类牛乳中常见的抗生素残留。检测76份未知原料乳样,检测结果与国外的试剂盒（如ECLIPSE50）一致。说明该抗生素检测微孔板可应用于牛乳样品的检测并获得可靠的检测结果。      

嗜热脂肪芽孢杆菌检测牛乳中抗生素残留

采用嗜热脂肪芽孢杆菌作为指示菌,制备牛乳抗生素检测微孔板,对牛乳中抗生素残留进行检测。结果表明,研制出的抗生素检测微孔板稳定性良好,操作简便、快捷、灵敏度高,结果容易判断。具有广谱性,可检测β-内酰胺类、磺胺类、四环素类和氨基糖苷类等多类牛乳中常见的抗生素残留。检测76份未知原料乳样,检测结果与国外的试剂盒(如ECLIPSE50)一致。说明该抗生素检测微孔板可应用于牛乳样品的检测并获得可靠的检测结果。     

牛奶中β-内酰胺类抗生素残留检测试剂盒的研制

以牛奶中残留的β-内酰胺类抗生素为检测目标,开发一种能快速检测其残留的试剂盒。利用嗜热脂肪芽孢杆菌BS 10208对酸和β-内酰胺类抗生素敏感的特点,优化其最优的生长条件,并制成在2.5 h内能发生显色反应的检测试剂盒。单因素结果表明,当菌液接种量5%、生长初始pH 8.0、培养温度55℃、蛋白胨添加量0.50%,其菌落数对数值分别为2.9、7.5、7.3、5.8。正交结果表明,在外加入0.5%蛋白胨营养肉汤中,接种5%种子液,在65℃恒温振荡器中培养24 h,其菌落数对数值为7.6。优化后检测试剂盒在2.5 h内能检测牛奶中β-内酰胺类抗生素残留,其各项参数为:反应体积为0.5 mL,1.5%琼脂,0.01 g/L溴甲酚紫浓度,BS 10208细胞总数为1.30×108 cfu,最适反应温度65℃,青霉素钠的限值为2～4μg/L。该试剂盒还能检测牛奶中残留的四环素类、磺胺类、大环内酯类、氨基糖苷类等抗生素。     

牛奶中抗生素残留检测的研究进展及应用现状

论述牛奶中残留抗生素的原因,列举商品化牛奶中抗生素残留的快速检测方法和试剂盒.主要的检测方法有微生物检测法、理化检测法、免疫法,并介绍其检测原理.     

嗜热脂肪芽孢杆菌检测牛奶中抗生素残留的研究

应用包埋的嗜热脂肪芽孢杆菌芽孢检测牛奶中的抗生素残留,对包埋中的营养物质、指示剂添加量以及包埋等条件进行了研究。结果表明,此方法对青霉素的检测极限值为0.003IU／mL,对阿莫西林抗生素检测极限值为4μg／kg。其灵敏性优于TTC法。     

牛奶中抗生素残留及其检测技术的研究进展

论述了牛奶中抗生素残留的原因和危害性,列举了目前国内外较流行的牛奶中抗生素的检测技术,并阐述了其检测原理。      

牛奶中抗生素残留及其检测技术的研究进展

论述了牛奶中抗生素残留的原因和危害性,列举了目前国内外较流行的牛奶中抗生素的检测技术,并阐述了其检测原理.     

牛奶中抗生素残留检测方法的研究现状

综述了抗生素残留的来源、危害，以及检测抗生素残留的各种方法，包括微生物检测法、理化检验法、免疫法等，指出应用免疫法快速检测试剂盒或生物传感器是今后抗生素残留控制与监督的发展方向．     

3种方法检测牛奶中抗生素残留

长期使用抗生素会造成畜禽免疫力下降,引起畜禽内源性感染和残留,而食用过抗生素超量的畜产品的人,会产生抗药性,或大量蓄积而对机体产生毒害。近年,牛奶中的抗生素残留成为人们日益关注的话题。     

Detection limits of four antimicrobial residue screening tests for β-lactams in goat's milk

This study was conducted to compare the detection limits (DL) of several antibiotic residue screening tests with the maximum residue limits (MRL) authorized by the EU according to the guidance for the standardized evaluation of microbial inhibitor tests of the International Dairy Federation. Composite antibiotic-free milk samples from 30 primiparous Murciano-Granadina goats in good health condition were used to prepare test samples spiked with different concentrations of each antimicrobial. In total, 5,760 analytical determinations of 10 β-lactam antibiotics (penicillin-G, ampicillin, amoxicillin, cloxacillin, oxacillin, dicloxacillin, cefadroxyl, cefalexin, cefoperazone, and cefuroxime) were performed using 4 antibiotic residue screening tests: the brilliant black reduction test BRT AiM (AiM-Analytik in Milch Produktions-und Vertriebs GmbH, München, Germany), Delvotest MCS (DSM Food Specialties, Delft, the Netherlands), Eclipse 100 (ZEU-Inmunotec SL, Zaragoza, Spain), and the Copan Milk Test (CMT; Copan Italia SpA, Brescia, Italy). For each method, we estimated the detection limits of the antimicrobial agents using a logistic regression model. Using the CMT and Delvotest on samples spiked with the 8 antibiotics for which MRL were available, DL were at or below the MRL. The BRT test provided DL at or below the MRL for all of the agents except cefalexin, whereas the Eclipse 100 method failed to detect 4 antibiotics (ampicillin, amoxicillin, cloxacillin, and cefoperazone) at MRL or below. Logistic regression-determined levels of agreement were highest for the CMT method (98.6 to 100%) and lowest for Eclipse 100 (66.3 to 100%). In general, agreement levels indicated good correlation between observed results and those predicted by logistic regression. The lowest b values (closely related to test sensitivity) were recorded for the cephalosporins (0.074 to 0.430) and highest for penicillin G, ampicillin, and amoxicillin (11.270 to 11.504). Delvotest and CMT best fulfilled IDF criteria for the ideal test for detecting antibiotic residues in milk.     

电子鼻对羊奶中三种抗生素残留的快速检测

目的 实现羊奶中常用抗生素残留的快速检测。方法 以电子鼻作为快速无损的检测仪器, 将含有不同浓度的青霉素G、硫酸庆大霉素、硫酸链霉素的羊奶作为研究对象, 利用线性判别分析(Linear Discriminant Analysis, LDA)和多层感知器神经网络(Multilayer Perceptron Neural Network, MLPN)等方法进行数据处理。结果 LDA的判别结果总体上能够将不同浓度的抗生素区分开来, 效果良好; MLPN的训练集和预测集的正确率也都在94.9%以上, 取得了很好的预测效果。结论 LDA 判定分组和MLPN预测模型可以用于羊奶中不同浓度青霉素G、硫酸庆大霉素、硫酸链霉素的快速检测。     

Short communication: Rapid antibiotic screening tests detect antibiotic residues in powdered milk products

Rapid antibiotic screening tests are widely used in the dairy industry to monitor milk for the presence of antibiotic residues above regulated levels. Given the persistent concern over contamination of milk products with antibiotic residues, we investigated the utility of IDEXX Snap test devices (IDEXX Laboratories Inc., Westbrook, ME) as tools for detecting antibiotic residues in powdered milk products. Five powdered milk products were reconstituted according to manufacturer specification with distilled water: Carnation (Nestlé USA Inc., Solon, OH), Nido youth and Nido adult (Nestlé Mexico Inc., Mexico City, Mexico), ELK (Campina, Eindhoven, the Netherlands), and Regilait (Saint-Martin-Belle-Roche, France). Positive samples were generated by spiking reconstituted milk with penicillin G, cephapirin, or tetracycline to either the European Union-regulated maximum residue limit or the FDA-regulated safe/tolerance level, whichever was lower. Control, unspiked negative milk samples and positive samples were tested with appropriate IDEXX Snap test kits (penicillin G and cephapirin with New Beta-Lactam, tetracycline with New Tetracycline). All samples yielded definitive results consistent with expectations, and there were no instances of false-positive or false-negative readings. These results suggest that both the New Beta-Lactam and New Tetracycline IDEXX Snap test kits effectively detect antibiotic residues in commercially available powdered milk samples and are useful tools for monitoring antibiotic residues in reconstituted powdered milk products.     

A microbiological inhibition method for the rapid,broad-spectrum,and high-throughput screening of 34 antibiotic residues in milk

In this study, we developed a microbiological inhibition method for the rapid screening of antibiotics in milk with Geobacillus stearothermophilus ATCC12980 as an indicator bacterium and an easy sample pretreatment. We observed that the limits of detection of the kit for 34 common antibiotic residues in milk, including β-lactams (13), aminoglycosides (6), tetracyclines (4), sulfonamides (6), macrolides (4), lincosamides (1), were lower than or close to the maximum residue limits formulated by the European Union and China. Moreover, the false-positive rate was 1% and the false-negative rates were less than 5%. The ruggedness of the method (the reproducibility of detection capability of different batches of medium) met requirements at determined levels and residual limits. The shelf life of the kit was more than 6 mo at 4°C. Additionally, we observed good correlations between the kit results and ultra-high-performance liquid chromatography-tandem mass spectrometry results for incurred milk (samples taken from animals treated with antibiotics according to the pre-slaughter medication data), which indicated that the kit was reliable for screening antibiotics in incurred samples. In conclusion, the kit has a broad application potential with high sensitivity, specificity, and reproducibility, stability, and reliability, combined with simple operation, low cost, and high-throughput capacity.     

Occurrence of antibiotic residues in milk from Manchega ewe dairy farms

Ewe milk samples from different ovine dairy farms from the Castilla-La Mancha region of Spain were collected from bulk tanks to estimate the occurrence of antibiotic residues in raw and heated (82 degrees C, 10 min) milk by the Delvotest SP test. The month of collection, somatic cell counts, and bacteriology were analyzed and examined by means of a logistic regression model. The screening of a total of 2686 raw milk samples showed 1.7% "positive" and 2.1% "doubtful" results, which decreased after heating treatment to 1.3% and 0.4%, respectively. "Positive" and "doubtful" samples were identified by penicillinase and p-aminobenzoic acid solutions, and the majority of them corresponded to antimicrobials different than beta-lactams or sulfonamides. By applying a logistic regression model, a significant effect of month of collection and bacteriology was observed in the initial screening and after the heat treatment. The highest percentages of "positive plus doubtful" results were observed in late summer-early autumn. A slight peak was also observed in spring in raw milk samples. Bacteriology was positively correlated with "positive plus doubtful" results.     

Evaluation of a microbiological indicator test for antibiotic detection in ewe and goat milk

Antibiotics are widely used for therapeutic and prophylactic purposes in dairy animals. The presence of residual antibiotics in milk could cause potentially serious problems in human health and have technological implication in the manufacturing of dairy products. The aim of this study was to evaluate Delvotest Accelerator (DSM Food Specialties, Delft, the Netherlands), a new system for a fully automated microbial test to detect antibiotic residues in ewe and goat milk. Forty-three samples of raw, whole, refrigerated bulk-tank milk samples (22 of ewe milk and 21 of goat milk) were analyzed during the whole lactation period. Four concentrations of 4 antibiotics were diluted in milk: penicillin G at 1, 2, 3, and 4 μg/L; sulfadiazine at 25, 50, 100, and 200 μg/L; tetracycline at 50, 100, 200, and 400 μg/L; and gentamicin at 25, 50, 100, and 200 μg/L. The detection limit of the Delvotest Accelerator was calculated as the range of antibiotic concentrations within which 95% of positive result lie. The range of detection limit of penicillin G and sulfadiazine was easily detected by Delvotest Accelerator at or below the European Union maximum residue limits, both for ewe and goat milk samples. In contrast, the system showed a lower ability to detect tetracycline and gentamicin both for ewe and goat milk samples. Very low percentages of false-positive outcomes were obtained. Lactation phase did not seem to be a crucial factor affecting the ability of the Delvotest Accelerator to detect spiked milk samples. A higher detection ability was observed for goat milk samples compared with ewe milk samples. A negative correlation between the percentage of positive milk samples detected and milk fat, protein, and lactose contents was observed for gentamicin only.     

表面增强拉曼散射基底的制备及其在农药 残留检测中的应用

表面增强拉曼散射光谱技术是探测界面特性、分子间相互作用和分子结构的一种高灵敏度的分析检测技术。近年来,此项检测技术在农药残留检测中得到广泛应用,其中增强基底发挥重要作用。本研究分析了表面增强拉曼散射增强基底制备方法的优缺点,并对表面增强拉曼散射光谱在农药残留检测中的应用进行综述。选择稳定性高的增强基底,结合样品前处理技术和融合其他光谱技术,提升农药残留的检测效率,提高检测准确度和灵敏度将是表面增强拉曼光谱技术在农药残留检测方面的发展方向。     

牛奶及奶制品中雌激素残留和检测的研究

牛奶在人类日常饮食结构中占有重要位置。近年来随着对环境雌激素研究的深入,牛奶中的雌激素问题越来越引起人们的关注。有研究表明,现代商业牛奶中雌激素含量显著上升。本文阐述了雌激素的分类及应用,并对牛奶和奶制品中雌激素残留状况,可能危害及检测方法进行了综述。     

Performance of blue-yellow screening test for antimicrobial detection in ovine milk

Drug residues in milk are important because of public health and industrial implications. The detection limits of 25 antimicrobial agents were determined by the blue-yellow screening method in ovine milk. For each drug, 8 concentrations were tested on 20 ovine milk samples from individual ewes in midlactation. Detection limits determined by means of logistic regression were below European Union maximum residue limits (EU-MRL) for penicillin G (3 to 4 μg/kg), ceftiofur (96 to 107 μg/kg), framycetin (720 to 781 μg/kg), neomycin (915 to 1,084 μg/kg), and tylosin (44 to 51 μg/kg). Detection limits for ampicillin (5 to 6 μg/kg), cloxacillin (33 to 42 μg/kg), cefoperazone (73 to 82 μg/kg), cefalexin (160 to 202 μg/kg), gentamycin (355 to 382 μg/kg), streptomycin (3,063 to 3,593 μg/kg), tilmicosin (109 to 131 μg/kg), erythromycin (444 to 522 μg/kg), spyramicin (1,106 to 1,346 μg/kg), sulfadimethoxine (101 to 119 μg/kg), sulfathiazole (122 to 151 μg/kg), sulfamethazine (309 to 328 μg/kg), sulfanilamide (1,750 to 2,674 μg/kg), tetracycline (233 to 257 μg/kg), oxytetracycline (398 to 501 μg/kg), doxycycline (323 to 419 μg/kg), chlortetracycline (3,331 to 3,989 μg/kg), danofloxacin (4.7 to 5.5 mg/kg), enrofloxacin (41 to 46 mg/kg), and flumequin (63 to 71 mg/kg) were higher than the EU-MRL. Although the blue-yellow method showed improved sensitivity compared with other tests studied in ovine milk, the performance of screening methods for detecting antimicrobial agents in milk of this species should be improved.     

Simultaneous determination of chlortetracycline,ampicillin and sarafloxacin in milk using capillary electrophoresis with electrochemiluminescence detection

ABSTRACT

A fast, inexpensive and sensitive approach for the simultaneous determination of chlortetracycline, ampicilline and sarafloxacin in milk was developed using capillary electrophoresis coupled with an electrochemiluminescence detector. Under the optimal detection conditions, the linear ranges for chlortetracyline, ampicilline and sarafloxacin were 0.030–5.0, 0.050–5.0 and 0.0040–2.0 μg ml^–1, respectively. The correlation coefficients of chlortetracycline, ampicilline and sarafloxacin were determined as 0.9997, 0.9952 and 0.9978, respectively. Detection limits (S/N = 3) of chlortetracycline, ampicilline and sarafloxacin were found as 0.017, 0.018 and 0.0013 μg ml^–1, respectively. This method was successfully applied for the determination of chlortetracycline, ampicilline and sarafloxacin in milk. The recoveries were between 95.3% and 100%. The relative standard deviations of the detection limit and recovery were less than 2.6% and 3.2%, respectively.