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1.
为了防止牛海绵状脑病(俗称疯牛病)和羊痒病传入我国,对来自疫区国家的动物源性饲料进行动物种类鉴别非常必要。本研究根据牛羊特异性卫星DNA建立了同一PCR体系同时检测羊、牛源性物质的方法。应用DNA提取液制备模板,不但有效地缩短了试验时间,还大大降低了成本,有利于在生产实际中推广应用。  相似文献   

2.
用分子生物学方法鉴别检测动物源性饲料中的牛羊源性成分   总被引:10,自引:0,他引:10  
为了防止海绵状脑病疫区和痒病疫区反刍动物源性饲料进入我国 ,非常有必要对贸易往来中的肉骨粉品种来源进行鉴定检测。我们采用分子生物学方法从动物源性饲料中鉴别检测牛、羊特异性线粒体DNA的片段 ,并用限制性内切酶SspⅠ对PCR产物进行酶切鉴定及DNA序列测定。该方法的灵敏度可达 0 .12 5 % ,具有灵敏度高、快速和特异性强的优点。  相似文献   

3.
应用线粒体DNA鉴定动物源性饲料成分研究进展   总被引:5,自引:0,他引:5  
为避免牛海绵状脑病和羊痒病疫区的蛋白饲料进口造成畜牧业上的经济损失,世界各国纷纷开展用分子生物学方法检测牛、羊源性蛋白饲料,本文就运用分子生物学方法有效检测动物物种间线粒体DNA特异性以确认动物源性蛋白饲料成分的来源作出阐述。  相似文献   

4.
商品中动物源性成分检测方法的研究进展   总被引:3,自引:0,他引:3  
动物源性成分的检测是识别商品掺假和防止疯牛病传播的有效途径.文章综述了目前动物源性成分检测方法的研究现状,对用于动物源性成分检测的电泳、色谱、光谱、免疫分析以及DNA检测等几种主要方法进行了评述和比较,并对该类项目检测方法的使用原则提出了建议.  相似文献   

5.
PCR方法鉴别肉骨粉中的动物成份种类   总被引:9,自引:0,他引:9  
为防范疯牛病传入我国,有必要鉴别肉骨粉中动物成份的种类。针对牛、羊、猪和鸡四种动物的特异性核苷酸序列,分别选用和设计了四对特异性引物,用试剂盒提取四种动物的肉骨粉或者处理过的肉组织中的DNA,然后进行PCR扩增,所扩增的目的片断大小分别为271bp、199bp、196bp、148bp,运用PCR技术建立了鉴定这四种动物源性成分的方法。结果分别扩增出四种动物的特异性条带,证明该PCR方法具有很高的特异性和敏感性,而且成本低廉,简便易行,因此可以作为鉴别肉骨粉种类的常规方法。  相似文献   

6.
饲料中鱼源性成分真实性鉴别研究   总被引:2,自引:0,他引:2  
研究旨在建立标准化的饲料中鱼源性成分的PCR检测方法,以形成行业标准推广使用.应用鱼源性引物对35种鱼类、24种非鱼类动物和10种植物样品进行PCR检测,只在35种鱼类中出现224 bp特异扩增条带,在其他的非鱼类动植物DNA中未出现扩增条带.实验表明,PCR检测方法具有特异性,检测限为0.1 ng DNA和0.1 %(W/W).应用该方法对上海口岸进口的100个鱼粉样品进行检测,均能检出鱼源性成分.研究建立的标准化饲料中鱼源性成分PCR检测方法能应用于日常检验检疫工作,可推广使用.  相似文献   

7.
PCR方法鉴别肉骨粉中的动物成份种类   总被引:2,自引:0,他引:2  
为防范疯牛病传入我国,有必要鉴别肉骨粉中动物成份的种类。针对牛、羊、猪和鸡四种动物的特异性核苷酸序列,分别选用和设计了四对特异性引物,用试剂盒提取四种动物的肉骨粉或者处理过的肉组织中的DNA,然后进行PCR扩增,所扩增的目的片断大小分别为271bp、199bp、196bp、148bp,运用PCR技术建立了鉴定这四种动物源性成分的方法。结果分别扩增出四种动物的特异性条带,证明该PCR方法具有很高的特异性和敏感性,而且成本低廉,简便易行,因此可以作为鉴别肉骨粉种类的常规方法。  相似文献   

8.
流行病学调查证明,用患有疯牛病和羊痒病的牛、羊等动物的肉和骨头制成的肉骨粉喂养动物会导致疯牛病和羊痒病的传播,是引起疯牛病发生的最初原因。目前,用于检测饲料中牛、羊源性成分的主要方法有:以检测骨头为基础的显微镜方法;以检测蛋白质为基础的免疫学方法和以检测DNA为基础的检测方法;其它方法还包括近红外光谱法等。  相似文献   

9.
为了对麝属动物源性成分进行有效鉴定,根据麝属动物线粒体DNA(mt DNA)序列,使用分子生物学软件Oligo7.0设计了特异性引物,用来建立PCR检测麝属动物源性成分的方法。结果显示:使用麝特异性引物优化的PCR体系可以对两份麝样品进行扩增,凝胶电泳均出现特异性条带。方法特异性好,只扩增麝属动物样品,16份其他物种的样品均为阴性结果。方法灵敏度较高,可检测到100个拷贝DNA的数量级。表明本研究建立的方法可应用于动物组织及其加工产品中的麝属动物源性成分的检测。  相似文献   

10.
为鉴定和区分饲料及动物产品中牛、山羊、绵羊源性成分,根据线粒体DNA(mitochondrial DNA,mtDNA)种间保守序列,设计合成了3对特异性引物与TaqMan探针,通过对荧光PCR反应体系和反应条件的优化筛选,建立了三重荧光PCR方法,在同一个荧光PCR反应中完成3种动物源性成分的检测。用该方法对16种不同源性的动物DNA进行检测,结果表明能特异地鉴别检测出牛、山羊和绵羊源性成分,且敏感性比现行国标PCR法高100倍。该方法适用于饲料、肉制品、奶制品等动物源性产品的检测。  相似文献   

11.
Specific conditions and practices of cattle feeding in Germany have to be taken into account for assessing the risk of feed born transmission of bovine spongiform encephalopathy, especially regarding the situation before the year 2000 when specific directives were introduced for feed production. The present retrospective epidemiological study includes data on feed production and the estimated amount of animal derived feedstuffs for the production of compounded feed for cattle. Risk assessment was performed based on the 'reproduction rate' (R0), that is defined as the estimated number of infections resulting from the processing of brain and spinal cord of BSE affected cattle that is recycled to bovines via feed. Under the conditions as given in Germany until the year 2000 the reproduction rate of BSE via the inclusion of animal derived feedstuffs in compounded feed production for cattle was estimated to be 1.1. Thus, it can be expected that BSE could be reproduced in the system, but with comparatively low efficiency. The expected incidence of BSE should be considerably lower compared to the situation during the 90th in the UK, due to the markedly lower recycling rate of animal protein in cattle feeding. Animal fat could have been a significant factor for BSE transmission due to contamination by proteinaceous brain and spinal cord material during the production process. The relative significance of fat containing feedstuffs for BSE transmission could have been higher in Germany compared to the situation in the UK where meat and bone meal was produced under different conditions and frequently used in higher proportions as an ingredient for compounded feed for ruminants.  相似文献   

12.
[目的]检测反刍动物饲料和动物源性饲料中牛羊源成分,掌握动物饲料安全情况。[方法]利用聚合酶链式反应法(PCR法)对反刍动物饲料产品(预混料、精料补充料、全价配合料等)和动物源性饲料产品(鱼粉、肉粉等)共计300个样品进行了牛羊源成分检测。[结果]牛羊源性成分检测合格率为99.33%。其中,反刍动物饲料产品260批次,牛羊源性成分合格率为99.23%;动物源性饲料产品40批次,均未检出牛羊源性成分。2个阳性样品均为牛源阳性。[结论]反刍动物饲料产品存在潜在的安全隐患。  相似文献   

13.
Nucleotide sequences of 16S rDNA and rpoB gene of 25 bovine and 6 ovine Histophilus somni strains were determined to detect subtle differences between the host animal species. The 1465 nucleotide residues of the 16S rDNA exhibited levels of sequence similarities of 99.4% or more. The high sequence similarity of the 16S rDNA of recently described species H. somni was confirmed in the 31 strains from cattle and sheep. These results suggested that the intra-specific diversity of 16S rDNA was limited in bovine and ovine strains of H. somni. The specific association of strains was also observed in the 311 bp region of rpoB gene which sequence similarities were 98.6% or more. However, the phylogenetic tree analysis of the rpoB gene showed that the ovine strains appeared to form a subgroup recovered in 70% of the bootstrap trees. In the 311 bp region of the ovine strains, a HincII restriction endonuclease site was detected. The PCR-amplified rpoB DNA of 46 bovine and 20 ovine H. somni strains were examined for the digestion with HincII. As the results, 17 strains of ovine strains were cleaved by the enzyme but none of the bovine strains appeared to possess the restriction site. The restriction enzyme analysis of rpoB gene may be useful to differentiate ovine strains from bovine strains of H. somni.  相似文献   

14.
The occurrence of BSE cases in Germany after the ban of meat and bone meal for ruminant feed in 1994 requires a detailed investigation of animal derived feedstuffs regarding their specific risks as vectors for the disease. Accepting the theory that BSE is a prion transmitted disease, the theoretical infectious potential was calculated for animal derived feedstuffs. This calculation was based on the assumption, that risk material (brain, spinal cord) of one clinically diseased cattle was rendered in the process as established in Germany (133 degrees C, 3 bar, 20 min) or, alternatively, that one diseased animal was slaughtered resulting in normal processing of the by-products for human food production. From this risk assessment it became obvious that meat and bone meal was one, but probably not the most important source for the spreading of BSE. Taking into account the high sensitivity of calves it can be speculated that certain products, e.g. from bone processing (bone meal) and fat melting (mixed animal fats), commonly used for the formulation of milk replacers, might have been more important as pathways. As it can't be excluded retrospectively that infected meat and bone meal was imported from the UK, this non-calculable influence may have been related to the significance of the other products. The calculation model underlines that efficient removal of specified risk material (brain, spinal cord) and adequate processing (133 degrees C, 3 bar, 20 min) or alternatively other equivalent treatments of fats are prerequisites for minimising the risk of feed borne transmission of BSE by animal derived feedstuffs. The epidemiological consequences are part of a subsequent paper.  相似文献   

15.
The leukotoxin of Mannheimia haemolytica has a very high degree of amino acid diversity because the lktA gene has a complex mosaic structure that has been derived by horizontal DNA transfer and intragenic recombination. The objective of the present study was to determine the effect of this amino acid diversity on leukotoxin cytotoxicity against bovine and ovine cell types. This was done by comparing the luminol-dependent chemiluminescence response of bovine and ovine neutrophils after pre-incubation with the various leukotoxin types. The LktA1.1-type leukotoxin associated with bovine serotype A1 and A6 strains differs from the LktA1.2-type leukotoxin produced by ovine serotype A1 and A6 strains at a single amino acid position and has enhanced activity against bovine but reduced activity against ovine neutrophils. These findings, together with the exclusive association of the LktA1.1-type leukotoxin with bovine strains, suggest that this leukotoxin type has an adaptive advantage in the bovine host. Leukotoxins LktA6-LktA10 are associated with ovine strains and have complex mosaic structures and diverse amino acid sequences but similar levels of cytotoxic activity against bovine and ovine neutrophils, respectively. However, ovine neutrophils were more sensitive to the cytotoxic activities of these leukotoxins than were bovine neutrophils. LktA8- and LktA10-type leukotoxins are associated with serotype A2 and A7 strains that are responsible for the majority of ovine disease cases, but LktA6-, LktA7- and LktA9-type leukotoxins are associated with less common serotypes. These findings contribute to the growing body of evidence suggesting that factors other than leukotoxin cytotoxicity are responsible for the full expression of virulence in M. haemolytica. Overall, the extensive recombinational exchanges within the lktA gene of M. haemolytica have had little effect on leukotoxin function which is highly conserved.  相似文献   

16.
动物产品中牛、羊源性成分多重PCR检测方法的建立   总被引:16,自引:2,他引:14  
以肉骨粉、鱼粉、猪肉干和鱼肉干为研究对象,异硫氰酸胍法提取总DNA,18S rDNA片段的扩增结果表明提取到的DNA中不存在抑制PCR的物质。应用梯度PCR技术对牛、羊源性成分检测的退火温度进行了优化,在单一PCR检测技术的基础上分别进行了18S rDNA片段和牛、羊源性成分的多重PCR分析,得到了预期的结果。试验表明,本文建立的多重PCR方法具有快速、简便、准确等特点,对动物产品牛、羊源性成分检测具有重要意义。  相似文献   

17.
For the veterinarian, drug sale is generally permitted by his licence of drug dispensing. According to the law on animal feed, feedstuffs are not regarded as drugs and thus they are not licensed to be sold by veterinarians. Drugs, however, may only be released for the treatment of individual animals, which are previously examined by the vet. The sale of animal feed is not regarded as part of veterinary practice. Only in case of a registered additional business, a veterinarian is allowed to sale feedstuffs. Dietetic animal feedstuffs are also regulated by the Animal Feed Act. Dietetic drugs, however, are regulated by the Drug Act and are strongly to be delineated from dietetic feed. Thus, the term "dietetic" alone does not automatically give any legitimatization for a sale of feedstuffs in veterinary practice.  相似文献   

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