免疫分析技术在乳及乳制品抗生素类兽药残留检测中的应用

乳牛饲养过程中,常使用抗生素类药物来预防和治疗乳牛疾病,但抗生素的大量应用,会使牛奶中含有大量抗生素,不仅降低牛奶的品质,还会给人体造成伤害。本文主要分析了免疫分析技术在乳以及乳制品抗生素类兽药残留检测方面的应用。     

乳与乳制品抗生素残留及其在病牛乳中持续时间的探讨

1986年6～7月对南京、无锡、苏州、常州、徐州、淮阴、泰州、盐城等市的10个乳牛场使用抗生素的情况和乳与乳制品中抗生素残留量及其持续时间进行了探讨。結果:各乳牛场均使用青霉素、链霉素等抗生素药物治疗乳牛的各种疾病。应用TTC法检测乳与乳制品中抗生素残留量的检出率时,生奶3.6%,消毒牛奶11.48％、奶粉39.58％;青链霉素水剂在常量下不论给药途径如何,奶中残留持续时间都不超过48小时。病侧乳室灌注抗生素后,在邻室分泌出的乳中不存在抗生素残留;肌肉注射抗生素,奶中残留经24小时就呈阴性,而乳室灌注法奶中的残留要经48小时才转阴性,因此提倡肌注法给药。     

牛奶中四环素类兽药残留的快速检测关键技术应用研究

乳制品的安全问题是关乎居民民生问题的重要食品健康问题。四环素类抗生素作为牛奶中的常用添加抗生素,其残留问题受到人们的日益关注。本文通过对目前国内外乳制品快速检测方法的分析,进一步探究牛奶中四环素类兽药残留的快速关键性技术,并通过试验的方式对四环素类的快速检测方式进行研究,以观察四环素的质谱峰值与混合液中四环素含量的整体情况,为探究快速、高效的的检测方法和检测技术提供参考性见解。     

牛奶中兽药残留检测前处理技术研究进展

随着我国经济的发展,国内乳制品的消费比例及需求不断增加,如何把好乳制品源头关-“牛奶”,成为了提高我国乳制品质量安全的重要环节之一。其中,由于在奶牛生产中,不合理使用兽药引起的兽药残留超标,成为影响消费者身体健康的重要隐患之一。在牛奶兽药残留检测过程中通常分为前处理和仪器分析两部分,由于牛奶基质复杂及兽药种类多、极性不一,使得在牛奶前处理过程中一方面要求尽可能地去除基质干扰,另一方面又要求尽可能地减少待测物流失。因此,对牛奶前处理提出了更高的要求。本文对国内外牛奶兽药残留检测中常用的前处理技术进行了综述,对其优缺点进行了分析比较,并对其发展方向进行了展望,以期为提高牛奶检测水平及质量安全提供帮助。     

酶联免疫法与乳及乳制品中抗生素残留的检测

对酶联免疫检测技术(ELISA)做了介绍,并对该技术在乳及乳制品抗生素残留检测的应用进行了评述.阐述了该项技术在乳及乳制品抗生素残留检测中的应用前景.     

液体乳中抗生素残留量快速检测方法探究

抗生素在畜牧业中广泛应用,不可避免地造成牛奶中残留抗生素.抗生素残留不仅危害人类健康,同时也影响牛奶的品质,造成经济损失.现检测生鲜乳中抗生素残留量方法众多.对3种快速检测方法(SNAP双流向酶联免疫法、嗜热链球菌抑制法(TTC)和ECLIPSE50试剂盒微生物抑制法)进行了比较,以寻求简便、快速、准确、敏感性高的方法,以满足日趋严格的残留限量要求,保障人们饮用牛奶的卫生和安全.     

浙江省部分地区鲜牛奶与消毒牛奶中氯霉素、四环素类抗生素残留的调查及检测分析

对浙江省范围内的鲜牛奶与消毒牛奶中氯霉素、四环素类兽药残留量进行专项调查,并用酶标免疫试剂盒(德国拜发试剂盒)对鲜牛奶与消毒牛奶中氯霉素、四环素类兽药残留进行检测。共调查了鲜牛奶和消毒牛奶各140批次,涉及53个厂家,10个地区,合格为132和136批次,不合格为8和4批次,合格率为94.3%和97.1%。建议有关卫生管理部门将抗生素检测纳入常规检测项目,以保证广大消费者饮用安全卫生的绿色食品。同时应特别加强对私营企业的管理和产品质量监测。     

乳和乳制品中抗生素残留的危害及检测方法

综述了乳与乳制品中抗生素残留现状和危害，比较了抗生素检测Delvotest法与其他几种方法。认为Delvotest法优于其他几种方法，建议有关部门尽快引进该技术，以加强我国乳和乳制品中的抗生素监测，解决目前乳业中面临的难题。     

内蒙古地方特色乳制品及含乳制品兽药残留调查

目的 了解内蒙古地方特色乳制品及含乳制品中兽药残留状况，为开展内蒙古地方特色乳制品及含乳制品兽药残留膳食暴露风险评估及食品安全地方标准制修订提供科学依据。方法 2021年开展了内蒙古地方特色乳制品中兽药残留监测工作，在12个盟市采集奶皮子、奶茶粉、嚼克等7类内蒙古地方特色乳制品及含乳制品共951份，其中散装样本370份，定型包装样本581份，采用超高效液相色谱-质谱联用方法对样本中喹诺酮类、四环素类、大环内酯类、磺胺类、β-受体激动剂类共5类35种兽药进行检测。结果 奶皮子、奶茶粉、嚼克、楚拉、含乳固态成型制品5类地方特色乳制品中检出兽药残留，检出率范围0.83%～3.62%；检出的兽药有萘啶酸、诺氟沙星、培氟沙星、磺胺邻二甲氧嘧啶、磺胺间甲氧嘧啶、磺胺苯吡唑、磺胺甲氧哒嗪和泰乐菌素，检出率范围是0.11%～1.37%；经统计学检验，散装样本兽药残留检出率与定型包装样本兽药残留检出率差异无统计学意义（χ2=0.484,P>0.05）。结论 尽管内蒙古地方特色乳制品中四环素类、磺胺类和大环内酯类兽药残留总体合格率较高（>99%），但仍存在兽药检出情况，建议相关部门进一步加强兽...     

牛奶中β-内酰胺和四环素类抗生素二联检测试纸条的初步研究

抗生素越来越多地被应用于畜牧业中,其在乳制品中的残留问题也备受关注.乳品中残留抗生素不仅危害人体的健康,而且影响经济贸易的正常进行.为适应基层检测牛奶中抗生素残留的需求,作者采用胶体金免疫层析方法,建立了可以同时测定牛奶中的β-内酰胺类抗生素和四环素类抗生素的二联胶体金试纸条.该方法简单可靠,可同时检测牛奶中12种β-内酰胺类抗生素和4种四环素类抗生素,是一种值得推广的定性筛选方法,可作为液相色谱等定量检测仪器法的补充.     

供港原料奶兽药残留危害及控制

讨论了原料奶中药物残留对人类健康以及乳制品加工造成的危害，介绍了中国大陆和香港两地关于原料奶中药物残留的相关规定，提出了控制供港原料奶中药物残留的5项措施，包括提高饲养和防疫管理水平、规范兽药管理、加强牛群管理、使用高效低毒的药物以及加强药残检测。     

原料乳中药物残留和掺假物质检测方法的研究进展

原料乳中药物残留或掺假物质的存在造成的安全事件屡屡发生,这使我国乳品行业面临着巨大的挑战。本文主要综述了常规检测法、免疫分析法、仪器检测分析法、生物传感器和蛋白质芯片技术等对原料乳中常见聚醚类抗生素、类固醇激素、苯二氮卓类药物、苯基脲类除草剂、四环素类抗生素和青霉素类抗生素等药物残留、物理性质与原料乳相近的物质、常见电解质、非电解质、防腐剂等掺假物质的检测方法,以及对单一成分掺假物和多种成分掺假物的检测方法,并比较了不同检测方法的准确度、灵敏度、成本等优缺点,以期为相关检测部门进行质量监测或生产厂家牛乳收购提供技术参考。     

生乳中体细胞数对乳品质量安全的影响研究进展

体细胞数增加会改变乳成分、影响牛乳风味及缩短乳制品货架期,并且使乳中抗生素和药物残留的风险增加。乳中体细胞数是衡量乳畜健康、乳品质量与安全的标准,也是国际判定隐形乳房炎的重要指标。为了保障乳品质量和安全,许多国家规定了生乳中体细胞数的限量。本文从体细胞数和乳品质量与安全的关系、不同国家对生乳中体细胞数的限量及影响体细胞数的因素和体细胞数的检测方法等方面作一综述。     

Multiclass methods for the analysis of antibiotic residues in milk by liquid chromatography coupled to mass spectrometry: A review

Milk is an important and beneficial food from a nutritional point of view, being an indispensable source of high quality proteins. Furthermore, it is a raw material for many dairy products, such as yoghurt, cheese, cream etc. Before reaching consumers, milk goes through production, processing and circulation. Each step involves potentially unsafe factors, such as chemical contamination that can affect milk quality. Antibiotics are widely used in veterinary medicine for dry cow therapy and mastitis treatment in lactating cows, which can cause the presence of antimicrobial residues in milk. In order to ensure consumers’ safety, milk is analyzed to make sure that the fixed Maximum Residue Limits (MRLs) for antibiotics are not exceeded. Multiclass methods can monitor more drug classes through a single analysis, so they are faster, less time-consuming and cheaper than traditional methods (single-class); this aspect is particularly important for milk, which is a highly perishable food. Nevertheless, multiclass methods for veterinary drug residues in foodstuffs are real analytical challenges. This article reviews the major multiclass methods published for the determination of antibiotic residues in milk by liquid chromatography coupled to mass spectrometry, with a special focus on sample preparation approaches.     

Novel in vitro systems for prediction of veterinary drug residues in ovine milk and dairy products

A new in vitro tool was developed for the identification of veterinary substrates of the main drug transporter in the mammary gland. These drugs have a much higher chance of being concentrated into ovine milk and thus should be detectable in dairy products. Complementarily, a cell model for the identification of compounds that can inhibit the secretion of drugs into ovine milk, and thus reduce milk residues, was also generated. The ATP-binding cassette transporter G2 (ABCG2) is responsible for the concentration of its substrates into milk. The need to predict potential drug residues in ruminant milk has prompted the development of in vitro cell models over-expressing ABCG2 for these species to detect veterinary drugs that interact with this transporter. Using these models, several substrates for bovine and caprine ABCG2 have been found, and differences in activity between species have been reported. However, despite being of great toxicological relevance, no suitable in vitro model to predict substrates of ovine ABCG2 was available. New MDCKII and MEF3.8 cell models over-expressing ovine ABCG2 were generated for the identification of substrates and inhibitors of ovine ABCG2. Five widely used veterinary antibiotics (marbofloxacin, orbifloxacin, sarafloxacin, danofloxacin and difloxacin) were discovered as new substrates of ovine ABCG2. These results were confirmed for the bovine transporter and its Y581S variant using previously generated cell models. In addition, the avermectin doramectin was described as a new inhibitor of ruminant ABCG2. This new rapid assay to identify veterinary drugs that can be concentrated into ovine milk will potentially improve detection and monitoring of veterinary drug residues in ovine milk and dairy products.     

表面增强拉曼光谱在乳和乳品有害物质检测中的研究进展

乳和乳制品作为人类必不可少的食品之一,其中的有害物质,如农兽药残留、真菌毒素、非法添加物质以及其他污染物对消费者的健康安全构成了严重威胁,同时也阻碍了乳制品行业的发展。表面增强拉曼光谱法（Surface Enhanced Raman Spectroscopy,SERS）作为一种新兴的检测方法,有望能够满足目前乳和乳制品高通量,高灵敏度的检测需求。本文总结了SERS方法在牛奶检测中的研究进展,包括金属纳米检测探针和SERS固相平台的制备、抗体和适配体等分子识别技术的应用、核酸扩增技术以及微流控技术等与SERS的结合。最后,对SERS在乳制品的研究发展方向和应用前景做了总结和展望。     

Characteristics of ripened Tronchón cheese from raw goat milk containing legally admissible amounts of antibiotics

The aim of this study was to evaluate the transfer of the most widely used antibiotics in dairy goats from milk to cheese as well as their effect on the cheese-making process and cheese characteristics during ripening. Antibiotic-free milk was spiked individually with 7 veterinary drugs (amoxicillin, benzylpenicillin, cloxacillin, erythromycin, ciprofloxacin, enrofloxacin, and oxytetracycline) at an equivalent concentration of the European Union maximum residue limit. Spiked goat milk was used to make mature Tronchón cheeses, which were analyzed at 0, 30, and 60 d of maturation to determine pH, chemical composition, proteolytic and lipolytic activities, and color and textural properties. A sensory evaluation of 60-d ripened cheeses was carried out. Cheeses from raw antibiotic-free goat milk were made simultaneously to be used as reference. The cheese-making process was unaffected by the presence of most antibiotics evaluated. Only erythromycin and oxytetracycline significantly increased the time required for cheese production (122 ± 29 and 108 ± 25 min, respectively). However, variable amounts of antibiotics, ranging from 7.4 to 68%, were transferred from milk to cheese, with oxytetracycline and quinolones showing the highest retention rates. In general, antibiotic residues present in the cheeses at the beginning of maturation decrease significantly along time. Thus, β-lactams and erythromycin residues were not detectable after 30 d of ripening. However, relatively high concentrations of enrofloxacin (148 ± 12 µg/kg) and ciprofloxacin (253 ± 24 µg/kg) residues were found in the cheeses after 60 d of maturation. The quality characteristics of the Tronchón cheeses were only slightly affected by such substances, with few significant differences in the free fatty acid concentration and color and textural properties of the cheeses. Results herein indicate that the use of goat milk containing antibiotics, such as quinolones, at the European Union maximum residue limit for cheese production could adversely affect the safety of the final products because relatively high concentrations of these substances could be retained in soft and semi-mature cheeses, making it necessary to assess the risk for consumer health. Studies on the partition of the antibiotic substances during cheese-making, using specific technologies, would be convenient to guarantee the safety of cheese and related products.     

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.