浙江省主要畜禽产区养殖废水中抗生素残留研究

结合固相萃取、高效液相色谱—串联质谱分析,建立了适合畜禽养殖废水中磺胺类、氟喹诺酮类、四环素类、氯霉素类等共29种抗生素残留的测定方法,废水样品经盐酸调节pH 6.0、离心后经HLB固相萃取柱净化,液相色谱—串联质谱正离子模式测定磺胺类、氟喹诺酮类、四环素类,负离子模式测定氯霉素类。空白地表水中磺胺类和氟喹诺酮类添加浓度为0.02～1.0μg.L-1,四环素类和氯霉素类添加浓度为0.04～1.0μg.L-1,回收率范围为68.6%～95.0%(n=5),相对标准偏差(RSDs)为2.8%～12.3%,方法的定量限(LOQ)为0.02～0.04μg.L-1。利用该方法对养殖废水、养殖场附近河域20个水样进行测定,检出5种磺胺类化合物和氟苯尼考。     

关于兽用抗菌药使用减量化行动的探索与实践

宁夏以兽药生产经营使用环节全链条监管、动物源细菌耐药性监测技术体系建立、养殖场生物安全体系构建、减抗替抗产品推广应用、示范点带动等重点工作为切入点,强化质量保障,重视创新驱动,构建“政府+企业”产学研合作机制,不断促进兽用抗菌药使用减量化,形成了一套“监管到位、技术保障、企业尽责、用药减量”的兽用抗菌药使用减量化技术推广模式。     

替代抗生素饲料添加剂研究进展

抗生素作为主要的饲料添加剂在几十年的广泛应用过程中造成了致病菌耐药性增强、动物免疫力降低、影响动物疫苗的效价、通过食物链影响人体健康等亟待解决的问题。我国农业农村部已发布公告,自2020年起我国饲料全面禁止添加使用抗生素类添加剂,饲料行业正式进入“无抗时代”。长期以来,研究人员积极寻求研发各种抗生素替代品及绿色无害的饲料添加剂,以消除或减轻禁用抗生素带来的一系列影响。就目前主要抗生素替代品的抗菌特点及抗菌机理进行综述,旨在为“禁抗”后抗菌性添加剂在饲料中的应用提供参考。     

Application of polyclonal and monoclonal antibodies for the detection of Xanthomonas campestris pv. campestris in crucifer seeds using immunofluorescence microscopy

Polyclonal and monoclonal antibodies (PCAs and MCAs) were tested for the detection ofXanthomonas campestris pv.campestris (Xcc) in cabbage seeds using immunofluorescence microscopy (IF). It was concluded that PCA 94, MCAs 20H6, 2F4, 18G12 and a mixture of MCAs 20H6, 18G12, 2F4 and 16B5 could be used to detect Xcc in seed extracts when 5 min and 2.5 h shaking of seeds are used as extraction methods. The reliability of confirming suspect colonies with MCAs and PCA 94 in IF depended in part on the seed lot tested and the antibody used. Some virulent Xcc strains derived from seed lots, did not react with MCAs 10C5, 2F4, 18G12, 17C12 and 16B5. On the other hand, saprophytic isolates obtained from one seed lot cross-reacted with MCA 17C12 and to a lesser extent with MCAs 2F4, 18G12 and PCA 94. No relationship was found between IF-reactions of Xcc strains using MCAs and reactions of Xcc strains in pathogenicity testing. Xcc andX. c. pv.amoraciae (Xca) could in general not be distinguished on the basis of reactions with MCAs and PCAs. Also in pathogenicity tests Xcc and Xca were hard to distinguish.     

Pharmacokinetics and Dosage Regimen of Enrofloxacin in Buffalo Bulls after Intramuscular Administration

The disposition kinetics and dosage regimen of enrofloxacin were investigated in breeding buffalo bulls following a single intramuscular administration of 5 mg/kg. The absorption half-life, half-life of the terminal phase, apparent volume of distribution and total body clearance were 0.262±0.099 h, 1.97±0.23 h, 0.61±0.13 L/kg and 210.2±18.6 ml/(kg.h), respectively. Therapeutic plasma levels (1 g/ml) were maintained for up to 6 h. A satisfactory intramuscular dosage regimen for enrofloxacin in buffalo bulls would be 8.5 mg/kg followed by 8.0 mg/kg at 8 h intervals.     

Endometrial Tissue Concentrations of Enrofloxacin after Intrauterine Administration to Mares

Endometritis in mares is a common cause of infertility. Conventional treatments of the disease have mostly been unsuccessful, so new therapeutic alternatives need to be investigated. This study evaluated the uterine disposition and plasma pharmacokinetic behaviour of a commercial formulation of enrofloxacin (EFX) given by the intrauterine (i.u.) route (2.5 mg/kg) in healthy mares. In order to evaluate the uterine inflammatory response, an initial histopathological study assessing polymorphonuclear cell infiltration was carried out in 20 mares over a 14-day period after treatment. In a second study, 6 healthy adult mares were used for the pharmacokinetic study. Samples of uterine tissue and plasma were collected from 0 to 24 h after the i.u. treatment with 5% EFX solution. Samples were analysed by conventional microbiological assay using an EFX-sensitive strain of Escherichia coli (ATCC 25922). There was a moderate but statistically nonsignificant inflammatory response following i.u. administration of either the formulation or the vehicle alone. Pharmacokinetic analysis of the uterine concentrations of EFX showed a slow and sustained depletion, with EFX remaining at concentrations above the MIC for 24 h after treatment. The area under the concentration–time curve obtained for the uterus suggested that EFX and its metabolites are specifically retained in the uterus, which is the target tissue for bacterial colonization. Neither study provided any evidence of EFX toxicity. In conclusion, these results are encouraging and suggest that EFX may be a useful local treatment in mares with bacterial endometritis.     

盐酸小檗碱与左氧氟沙星联合抑制MRSA及其机制研究

养殖场抗生素滥用造成多种耐药性细菌的产生,从奶肉类制品中分离出耐甲氧西林金黄色葡萄球菌（MRSA）的案例逐年增长,对养殖业食品安全造成巨大威胁。本研究筛选出与左氧氟沙星具有联合抑制MRSA效果的天然产物盐酸小檗碱（BBR）,探究BBR与其对MRSA的协同抑菌效果和联合抑菌机制。结果表明,BBR能有效抑制MRSA生长,提高MRSA对左氧氟沙星的敏感度,两者联用后,MRSA对左氧氟沙星的MIC降为之前的1/16。协同作用机制主要为上调ribA及下调mec A和msc L表达水平,破坏细胞壁、增加细胞膜的通透性,从而达到协同抑菌的目的。本研究有助于降低畜牧养殖中抗生素的使用,为提高肉奶类食品安全奠定基础。     

中链脂肪酸抗菌和诱导防御肽表达的功能及其在仔猪饲料中的应用

近年,越来越多的研究表明,中链脂肪酸（MCFAs）抵抗病原菌是哺乳动物先天防御系统的重要组成部分,而且MCFAs也能够诱导人、猪和鸡内源性防御肽的表达,MCFAs的这些新功能尚未引起重视。MCFAs还和饲用有机酸或饲用植物精油具有协同抗菌增效作用,可以减少这些活性物质的使用量。相比于长链脂肪酸,日粮中添加MCFAs能显著地提高动物机体内氧气消耗量和线粒体呼吸速率,但产生活性氧自由基少,特别适合幼龄动物肠代谢和肝代谢所需的快速能量供应特点。日粮添加低浓度MCFAs（0.1%—0.5%,质量比）能显著提高新生或断奶仔猪存活率、粗蛋白粗脂肪消化率、饲料转化率,调节肠道菌群和改善肠上皮结构,进而促进生长。基于MCFAs的上述功能,将MCFAs与饲用有机酸或植物精油复配制备成微囊颗粒,可能是其用于仔猪抗生素替代品的尚佳方式。     

青菜对三种土壤中典型抗生素的累积规律研究

为探究青菜对不同类型土壤中不同种类抗生素的吸收和累积特性,通过盆栽实验,观察了三类土壤（黄褐土、砂姜黑土、红壤）中磺胺二甲嘧啶（SM2）、磺胺甲噁唑（SMZ）、四环素（TC）、土霉素（OTC）4种典型抗生素在青菜中的累积规律。结果表明：青菜累积三类土壤中4种抗生素的含量均在第10天达到最高后逐渐下降;青菜中抗生素的含量随着土壤中抗生素初始含量（0.1~25.0 mg·kg^-1）的增大而增大,抗生素初始含量为25.0 mg·kg^-1时,青菜中累积的抗生素含量显著高于其他低浓度处理组（P<0.05）;土壤中抗生素初始含量为25.0 mg·kg^-1时,青菜从不同类型土壤中吸收同种抗生素的含量差异较大,排序为红壤（SM₂ 14 993.6μg·kg^-1、SMZ 12 199.2 μg·kg^-1、TC 646.1 μg·kg^-1、OTC 967.6 μg·kg^-1）>黄褐土（SM₂ 12 598.1 μg·kg^-1、SMZ 11 678.5 μg·kg^-1、TC463.5 μg·kg^-1、OTC 663.8 μg·kg^-1）>砂姜黑土（SM₂ 9 510.4 μg·kg^-1、SMZ 3 666.9 μg·kg^-1、TC 58.8 μg·kg^-1、OTC 90.5 μg·kg^-1）,土壤pH和有机质含量是影响青菜从土壤中累积抗生素的重要因素;在同类土壤中,青菜对不同抗生素的累积顺序为SM₂>SMZ>OTC>TC,导致青菜对不同抗生素累积差异的原因,除了土壤对四环素类抗生素（TCs）的吸附能力强于磺胺类抗生素（SAs）外,还与不同抗生素的理化性质（分子结构、形态）有关。青菜能吸收土壤中的抗生素,在移栽后第10天青菜中抗生素的含量最高,青菜易从酸性土壤（红壤）中吸收抗生素,中性土壤（黄褐土）次之,碱性土壤（砂姜黑土）最低,且青菜对SAs的累积能力强于TCs,土壤中抗生素的初始含量越高,青菜中抗生素的含量也越高。     

石墨烯对喹诺酮在饱和多孔介质中运移的影响

为研究石墨烯（Graphene,GN）对喹诺酮类抗生素在地下水中运移的影响,以环丙沙星（Ciprofloxacin,CIP）和诺氟沙星（Norfloxacin,NOR）作为两种典型的喹诺酮类抗生素污染物,通过批量吸附实验和砂柱实验研究GN对CIP和NOR在饱和多孔介质中运移的影响。吸附实验结果表明,GN对两种抗生素污染物均具有较好的吸附性能,GN对CIP和NOR的Langmuir最大吸附容量分别为270.68 mg·g^-1和178.36 mg·g^-1。砂柱实验结果表明：随着多孔介质中GN含量从10 mg增加到80 mg,CIP和NOR在一维砂柱中的迁移能力降低;随着流速和电解质浓度（Na⁺和Ca²⁺）的增大,回收率逐步升高,CIP和NOR的运移能力也逐步增强。根据BDST模型对不同条件下的CIP和NOR在一维砂柱中的运移过程进行了模拟和预测,模型对穿透时间的预测值与实验值接近,表明BDST模型能较好地预测多孔介质中GN对CIP和NOR迁移能力的影响。