抗球虫药物残留的免疫检测技术

抗球虫药物在畜禽业应用广泛,可预防治疗球虫病,提高家禽饲料转化率,提升肉品质。但是,随即会出现饲料交叉污染,对非目标动物产生毒性作用,动物性食品中药物残留超标等问题。因此,建立简便、快速、灵敏度高的检测方法检测动物源性食品中的抗球虫药物残留极为重要。基于抗原-抗体特异性结合原理的免疫分析检测技术能够完成快速检测,高通量筛选,具有特异性强、灵敏度高、操作简便、成本低的优点。本文综述了不同基质中抗球虫药物残留的免疫检测技术进展,重点介绍了酶联免疫吸附检测法、免疫层析法、荧光偏振免疫分析检测法、时间分辨荧光免疫分析检测法和生物传感器检测法。并对免疫检测技术在残留检测方面的发展趋势进行了展望,旨在为抗球虫药物的残留监控提供方法学上的参考,为新方法的建立提供思路。     

Treatment of contaminated radial fracture in Sprague-Dawley rats by application of a degradable polymer releasing gentamicin

Fracture-related infections remain a leading cause of morbidity and mortality. We aimed to establish a simple contaminated radial osteotomy model to assess the efficacy of a biodegradable polymer poly(sebacic-co-ricinoleic acid) [p(SA-RA)] containing 20% w/w gentamicin. A unilateral transverse osteotomy was induced in Sprague-Dawley (SD) rats, followed by application of Staphylococcus aureus suspension over the fracture. After successfully establishing the contaminated open fracture model, we treated the rats either systemically (intraperitoneal cefuroxime), locally with p(SA-RA) containing gentamicin, or both. Control groups included non-contaminated group and contaminated groups that were either untreated or treated with the polymer alone. After 4 weeks, the bones were subjected to micro-CT scanning and microbiological and histopathology evaluations. Micro-CT analysis revealed similar changes in the group subjected to both local and systemic treatment as in the non-contaminated control group. Lack of detectable bacterial growth was noted in most animals of the group subjected to both local and systemic treatment, and all samples were negative for S. aureus. Histopathological evaluation revealed that all treatment modalities containing antibiotics were highly effective in reducing infection and promoting callus repair, resulting in early bone healing. While p(SA-RA) containing gentamicin treatment showed better results than cefuroxime, the combination of local and systemic treatment displayed the highest therapeutic potential in this model.     

Pathological analysis of batch safety testing of veterinary vaccines using small laboratory animals

Batch safety tests (BSTs) of veterinary vaccines are conducted using small laboratory animals to assure the safety of vaccines according to several criteria, including clinical signs and change in body weight. Although the latter is used as an evaluation index in BSTs, there have been no reports on the internal changes that affect body weight during the test period. Therefore, we analyzed BST via pathological examination of the tested animals. Here, BSTs were performed for 176 batches using mice and 126 batches using of guinea pigs. Most of the gross findings could be classified into four lesion types (nodules, adhesions, ascites, no apparent signs), with only one vaccine inducing lesions that could not be classified into any of these four types. Histopathological examination revealed that the reactions caused by BST were pyogenic and/or granulomatous inflammation. Nodular or adhesive lesions comprised more severe pyogenic granulomatous inflammation than ascites or cases with no apparent gross lesions. These nodular or adhesive lesions were more frequently induced by vaccines that contained an adjuvant than by vaccines that did not contain an adjuvant. The cases with “exceptional” gross findings histologically presented severe necrosis of the hematopoietic system. Additional testing showed that these “exceptional” lesions were induced when a specific type of light liquid paraffin was injected along with other vaccine additives. Our results show that body weight loss and/or lesions during BST were induced by proinflammatory properties of the tested vaccines and that BST is a sensitive method for detecting unexpected effects of vaccine components.     

饲料中动物源性成分微流控检测芯片的研究

试验以"LAMP+微流控"技术为基础,开发饲料动物源性成分检测芯片。针对牛、羊、猪、鸡的16srRNA靶序列,根据LAMP引物设计软件进行4种肉类LAMP引物设计。在LAMP优化试验、芯片优化测试、特异性验证试验和芯片应用测试中,进行最佳引物筛选测试和特异性测试。结果显示,筛选出的4种肉类最佳引物特异性强,最佳反应温度为65℃。芯片特异性验证试验的归一化荧光曲线显示,试验结果准确、特异性稳定。对饲料市场购买的16份样本中检测出的动物源性成分,采用中华人民共和国农业行业标准验证,符合率100%。微流控检测芯片的开发与应用为饲料中动物源性成分检测提供新方法,为饲料产品质量安全提供保障。     

分散固相萃取结合UPLC-Orbitrap MS联用测定鸡蛋中五氯苯酚残留量

建立了超高效液相色谱-静电轨道阱质谱联用测定禽蛋中五氯苯酚残留量的分析方法。取均质后的禽蛋样品经乙腈溶液提取,加入无水硫酸镁和氯化钠进行脱水和盐析,高速离心分层后,取适量乙腈层溶液经0.1%(V∶V)甲酸稀释后,用酸性氧化铝粉末进行样品净化并以10000 r/min离心取上清液,过0.22μm滤膜后上机测定。使用反相色谱柱进行分离,流动相为乙腈:0.1%甲酸=75∶25,采用等度程序进行洗脱,静电轨道阱质谱进行定性定量分析。结果表明,五氯苯酚在0.5~100μg/kg范围内线性关系良好(r>0.995),方法的最低检出限为0.2μg/kg,最低定量限为0.5μg/kg,添加回收率在74.43%~96.85%之间,批内批间变异系数CV%<10%。该方法具有较好的准确度与精密度,适用于禽蛋中五氯苯酚残留量的测定。     

LC-MS/MS法分析生猪异位发酵床垫料中5大类兽用抗菌药物残留及蓄积趋势

建立了生猪异位发酵床垫料中四环素类、喹诺酮类、磺胺类、酰胺醇类和大环内酯类共54种抗菌药物残留量的LC-MS/MS检测方法,经超低温冷冻粉碎后的垫料样本,以费休氏-库仑滴定法测定水分,用Mcllvaine-Na2EDTA缓冲液和乙腈提取,经亲水亲脂平衡固相萃取小柱净化,用LC-MS/MS法测定抗菌药物残留量,最终结果以折水后含量表示。方法具有简便、高通量、抗干扰的特点,可以满足生猪异位发酵床垫料中主要兽用抗菌药物残留检测的要求。利用该方法,对12家养殖场垫料样本中5大类兽用抗菌药物的残留量进行了检测。12家养殖场垫料样本中共检出25种抗菌药物,各场垫料样本抗菌药物总残留量在0.7 g/t~28 g/t之间,其中四环素类药物对总残留量的贡献率在91.3%～98.8%之间。     

白酒糟饲料化开发利用及其对贵州石漠化地区的启示

根据国家石漠化综合治理草地畜牧业工程的需要,应用营养学、饲料学等学科理论,本文对贵州喀斯特石漠化地区白酒糟资源的利用价值进行了综合评价,包括其营养价值、饲用价值、生态价值和经济价值。同时针对白酒糟饲料化开发利用过程中存在的加工技术滞后、理论研究不足、地域条件限制等问题进行了分析,提出了运用青贮法和微生物发酵等关键技术、强化基础理论研究、构建饲料加工中心等相应解决对策,并指出大力发展以白酒糟为原料的非常规饲料加工业是喀斯特石漠化地区发展生态畜牧业的重要举措,也是巩固石漠化治理成果的有效办法。     

新型广谱杀虫药沙罗拉纳在国外伴侣动物体外寄生虫防治中的应用

沙罗拉纳(sarolaner)属异恶唑啉类化合物,是一种新型广谱杀虫药。沙罗拉纳已被证实通过特异性阻断昆虫γ-氨基丁酸受体和谷氨酸门控氯离子通道而显示出抗寄生虫活性。本文综述了近年来沙罗拉纳在国外伴侣动物体外寄生虫防治中的开发及临床应用情况,旨在为该药的深入研究和国内开发应用提供参考。近年来,该药主要用于犬猫等伴侣动物体外寄生虫的临床预防和治疗,对蜱类、蚤类、螨类等体外寄生虫有高效的杀灭作用。与其他杀虫药相比,该药在分子结构、作用位点和选择性等方面有显著差异,具有杀虫活性强、治疗剂量低、安全性高等优点。此外,该药通过与多种药物联合应用,可提高治疗效果,扩大治疗范围。就国外研究来看,沙罗拉纳是一种安全广谱的高效杀虫剂,拥有广阔的应用前景。     

《家禽屠宰检疫规程》中有关问题分析

《家禽屠宰检疫规程》在规范家禽的屠宰检疫,加强家禽产品的检疫活动管理,保障动物源食品安全方面发挥了重要作用。本文概述了《家禽屠宰检疫规程》关于家禽屠宰场所定义、规程适用动物、检疫对象、检疫合格标准的判定等方面存在的问题,特别是针对家禽产品入库贮藏后才出厂如何出具检疫合格证明,提出可采取贮藏后继续调运或分销的检疫方式出具检疫证明这一解决方案。这提示在今后工作中扩大屠宰检疫的适用范围及检疫对象,进一步修订和完善屠宰检疫规程,提升屠宰检疫技术水平。