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1.
Johne’s disease (paratuberculosis) is an economically important disease of cattle worldwide. The disease is caused by Mycobacterium avium subsp. paratuberculosis (MAP), a fastidious gram-positive bacterium. PCR is increasingly used in diagnostic laboratories for the detection of MAP in fecal samples given the rapid test turnaround time and sensitivity and specificity comparable to fecal culture. However, efficient extraction of DNA for sensitive detection of MAP by PCR is affected by the complex lipid-rich cell wall of MAP and the presence of PCR inhibitors in feces. We evaluated a high-throughput nucleic acid extraction method (MagMAX core nucleic acid purification kit with mechanical lysis module) in conjunction with an hspX gene PCR for the detection of MAP from bovine fecal samples, which resulted in correct identification of all negative (13 of 13) and positive (35 of 35) proficiency test samples obtained from the National Veterinary Services Laboratories. In addition, all 6 negative and 50 of 51 positive diagnostic specimens tested were categorized correctly.  相似文献   
2.
由副结核分枝杆菌引起牛的副结核病常导致感染牛的慢性增生性肠炎,以及体重、产奶量等生产性能下降,给畜牧业带来了严重的经济损失,目前尚无有效治疗药物。人们常采用不同的检测技术定期对牛群进行监测、淘汰活动带菌期的牛、对牛群进行疫苗接种、采用生物安全防控等措施比较有效地避免了副结核分枝杆菌在牛群中的传播。文章主要从副结核分枝杆菌背景,牛副结核病的危害及防控措施三个方面进行了综述,以期为牛副结核病的防控提供思路。  相似文献   
3.
Summary

Numerous epidemiological reports implicate foods of animal origin as vehicles of human campylobacteriosis. Pigs are probably an important reservoir of campylobacter and a potential source of human infection. In order to improve our knowledge of the epidemiology of campylobacter in pigs, the prevalence of campylobacter and its contamination of feed were monitored in eight pig farms. Faeces samples of pigs aged 11 and 22 weeks, and samples of rectal, ileal and gastric content at a slaughterhouse were collected for bacteriological examination. On 5 farms, subsequent groups of pigs housed in the same stalls was sampled, too. A selection of the campylobacter isolates was characterized with a genetic typing method (RFLP). More than 85% of the sampled porkers were shown to be intestinal carriers of campylobacter at all stages of fattening. Subsequent groups of pigs housed in the same stalls were all carriers, too. The level of campylobacters in the faeces tended to decrease as the pigs got older. There was no difference in the frequency and level of infection with campylobacter between porkers on different farms. The feeding system (wet feed versus dry pellets) did not seem to influence the prevalence of campylobacter although wet feed gave lower counts of Enterobacteriaceae in the faeces.

RFLP‐typing showed a high diversity of campylobacter strains at each sampling on the farm. Similarities were seen between strains isolated during two subsequent samplings of the same group of pigs, but not between strains isolated on the same farm from subsequent groups of pigs housed in the same stall. This suggests that the piglets were already infected at a young age on the breeding farm.  相似文献   
4.
为建立牛副结核分枝杆菌(MAP)的快速检测方法,本研究采用环介导等温扩增(LAMP)技术,以MAP的IS900基因序列为靶基因设计特异性引物,建立了MAP特异性的LAMP快速检测方法。经反应条件优化,检测结果显示,建立的LAMP检测方法具有良好的特异性,最低见测量为0.53 pg;与其他病原菌无任何交叉反应。临床样品检测与行业标准的符合率为100%。该LAMP检测方法可以用于MAP的常规检疫。  相似文献   
5.
OBJECTIVE: To investigate the development of immune responses in calves experimentally and naturally infected with Mycobacterium paratuberculosis and to evaluate the potential for diagnostic tests to detect infected calves. DESIGN: Sequential testing of four treatment groups of calves over a 2 year period. PROCEDURE: Twenty-nine calves were allocated to four groups. Group D calves were orally dosed with M paratuberculosis, group N calves naturally exposed to M paratuberculosis, group V calves vaccinated for M paratuberculosis, and group C were control calves (not infected or vaccinated). Blood and faecal specimens were collected from each calf at monthly intervals to 18 months of age and then every 2 months until they were slaughtered between the ages of 21 and 29 months. Specimens were tested using absorbed EIA, IFN-gamma EIA and faecal culture. The infection status of the calves was confirmed by extensive histopathological examination and tissue culture. RESULTS: M paratuberculosis infection was confirmed in 10 calves, comprising six of eight orally dosed calves, three of five naturally exposed calves and one of nine vaccinated calves. The six artificially infected calves and one naturally infected calf were detected shedding M paratuberculosis in their faeces. Results with positive absorbed EIA were obtained from one artificially infected calf, one naturally infected calf and three vaccinated calves. All calves including controls had positive results on at least one occasion using the IFN-gamma EIA. In addition, seven calves had positive bovine tuberculosis results using the IFN-gamma EIA, even though bovine tuberculosis has been eradicated from Australia. CONCLUSION: Detection of M paratuberculosis infection in young cattle continues to be difficult using current tests.  相似文献   
6.
为表达并纯化禽分枝杆菌副结核亚种主要抗原基因的串联重组蛋白r22-ag85B,期望研制出一种新型副结核病疫苗,以禽分枝杆菌副结核亚种参考株P18的基因组DNA为模板,扩增了22KD基因和ag85B基因。采用重叠延伸剪接PCR技术(SOE-PCR)获得了融合基因22-agS5B,将基因连接于表达载体pET32a(+),构建了重组质粒pET22-ag85B。将其转化到大肠杆菌感受态细胞BL21(DE3)中,以IPTG(终浓度1mmol/L)诱导后对表达产物进行Western blot检测。检测结果显示,成功表达并纯化了重组蛋白r22-ag85B,其分子质量约为65ku。Western blot检测表明此蛋白具有良好的免疫学活性。禽分枝杆菌副结核亚种22-ag85B重组蛋白的成功表达及纯化为副结核病疫苗的研究工作奠定了基础。  相似文献   
7.
微量补体结合试验检测牛、羊副结核病的研究   总被引:1,自引:0,他引:1  
采用《中华人民共和国出入境检验检疫行业标准》“副结核病补体结合试验操作规程”方法(简称常量法)和微量补体结合试验(简称微量法)检测3677头进口牛羊的副结核病抗体并进行比较。结果表明,常量法和微量法的溶血素效价测定值相同,补体效价测定值近似,阴性血清对照、抗原对照、补体对照、红细胞对照、血清抗补体结合对照结果均相同,阳性对照血清效价微量法比常量法提高1个滴度,被检血清的阳性检出率微量法高于常量法。比较37℃水浴及37℃恒温培养箱环境下的微量法补体结合试验结果,溶血素效价、补体效价、各项对照试验结果完全相同,被检血清在37℃水浴反应更加充分。比较用变态试验和补体结合试验检验3677头牛羊副结核病的结果、以及用补体结合试验、变态反应和酶联免疫吸附试验方法同时检测2024头牛的结果,发现它们的相关性很小。  相似文献   
8.
牛副结核检疫中的非特异性干扰   总被引:2,自引:0,他引:2  
给犊牛接种各种分枝杆菌,然后以副结核PPD进行变态反应试验,观察各分枝杆菌感染对副结核变态反应的干扰;从各种分枝杆菌感染牛群采取血清进行副结核ELISA检测,以观察各分枝杆菌血清对牛副结核ELISA的干扰。结果:多种分枝杆菌,特别是鸟胞内分枝杆菌、瘰疬分枝杆菌的感染可干扰副结核变态反应;部分结核牛血清干扰副结核ELISA的检测  相似文献   
9.
副结核分枝杆菌DNA探针的制备与应用   总被引:1,自引:0,他引:1  
以溶菌酶、SDS、高氯酸钠等处理提取副结核分析杆菌C2株染色体DNA,经限制性内切酶Pst I消化后,以质粒pBluescript SK为载体,通过T4DNA连接酶连接,转入E。coli DH5a受体菌中,构建了副结核菌C2株的DNA基因文库。应用反向杂交试验,从基因文库中筛选出4个重组克隆:PTP12、PTP19、PTP38。对这4个重组克隆进行酶切、电泳分析,结果表明4个插入片段长度分别为:4  相似文献   
10.
Molecular biology has contributed to our knowledge and understanding of the structure of Mycobacteriumavium subspecies paratuberculosis and has been particularly useful in determining those components that elicit immune responses in the host or discriminate M. avium paratuberculosis from other closely related environmental mycobacteria. As such, it has made a significant impact in the field of diagnosis, and has been instrumental in the development of specific and sensitive diagnostic tests. The next decade will see exciting new developments in paratuberculosis research as a consequence of substantial advances made in the construction of gene transfer systems in mycobacteria. These will provide opportunities for applying new strategies to determine the genetic basis for pathogenesis and the mechanisms of drug resistance and will offer new prospects for the rational design of efficient vaccines.  相似文献   
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