共查询到20条相似文献，搜索用时 945 毫秒
1.
为建立同时快速检测奶牛奶样中无乳链球菌、停乳链球菌、乳房链球菌和金黄色葡萄球菌的方法,根据无乳链球菌sip基因、停乳链球菌isp基因、乳房链球菌pauA基因和金黄色葡萄球菌nuc基因各设计1对特异性引物,建立多重PCR检测体系。结果显示,该检测方法具有高特异性,无乳链球菌、停乳链球菌、乳房链球菌和金黄色葡萄球菌敏感性分别为105、104、105、105 CFU/mL。对临床采集的460份奶样检测结果表明,建立的多重PCR体系可以用于临床上无乳链球菌、停乳链球菌、乳房链球菌和金黄色葡萄球菌感染引起的奶牛乳房炎的检测。 相似文献
2.
多重PCR快速检测奶牛乳房炎3种主要病原体 总被引：10，自引：0，他引：10
奶牛乳房炎是引起奶牛业经济损失的一种重要疫病,目前还没有快速、特异检测奶牛乳房炎主要致病原的方法。本试验根据金黄色葡萄球菌、无乳链球菌、大肠杆菌各自保守的16S或23S rRNA基因序列,合成了3对特异性引物,建立了三重PCR检测方法。特异性试验表明,该方法对所有参与测试的金黄色葡萄球菌、无乳链球菌和大肠杆菌都能扩增出各自的阳性条带,而对所有参与测试的对照菌株则不能扩增出任何条带。敏感性试验表明该方法能检测到4个菌的金黄色葡萄球菌、无乳链球菌和2个菌的大肠杆菌。对送检的乳房炎奶样36份直接进行PCR检测,金黄色葡萄球菌阳性7份,无乳链球菌阳性2份,大肠杆菌阳性6份。 相似文献
3.
4.
多重PCR检测奶牛乳腺炎金黄色葡萄球菌、无乳链球菌、停乳链球菌和酵母菌方法的建立与应用 总被引：2，自引：0，他引：2
参照GenBank发表的序列，在金黄色葡萄球菌、无乳链球菌和停乳链球菌16SrRNA与23SrRNA之间的区域设计了3对引物，参照念珠菌和隐球菌的18SrRNA的序列设计1对引物，建立了检测金黄色葡萄球菌、无乳链球菌、停乳链球菌和酵母真菌4种乳腺炎主要致病菌的多重PCR方法。参照Skladny的方法制备模拟了细菌感染l临床标本。结果表明：本试验建立的多重PCR方法具有较好的特异性，多重PCR方法检测乳样中的金黄色葡萄球菌的细菌最小浓度为10^4CFU／mL，检测无乳链球菌、停乳链球菌和酵母真菌的细菌最小浓度分别为10^4CFU／mL、10^3CFU／mL和10^3CFU／mL。通过对采自临床型乳腺炎（46个）和隐性乳腺炎（167个）动物共计213个乳样分别用传统细菌学培养法和多重PCR方法进行检测，多重PCR对金黄色葡萄球菌和酵母真菌的检测具有更高的检出率（P〈0．01），但该方法对无乳链球菌和停乳链球菌的检出率与培养法差异不显著（P〉0．05）。 相似文献
5.
《中国兽医学报》2019,(11):2195-2199
为同时检测奶牛源金黄色葡萄球菌、无乳链球菌和绿脓杆菌,本试验根据GenBank中3种病原菌的nuc、ef-tu和eta基因保守序列分别合成了3对特异性引物,通过对反应条件、反应体系优化,建立了多重PCR检测方法。特异性试验结果表明,该方法与大肠杆菌、克雷伯杆菌、表皮葡萄球菌、巴氏杆菌无交叉反应,敏感性试验结果表明,多重PCR检测方法对金黄色葡萄球菌、无乳链球菌和绿脓杆菌的检出限为10~(-3) mg/L。应用该方法对150份临床样品进行检测,结果金黄色葡萄球菌与无乳链球菌混合感染检出率为28.7%(43/150),金黄色葡萄球菌与绿脓杆菌混合感染检出率为14%(21/150),无乳链球菌与绿脓杆菌混合感染检出率为11.3%(17/150),3种菌混合感染检出率为6%(9/150)。应用多重PCR检测结果与分离培养诊断方法结果进行比较,均能检测出样品中的病原菌,2种检测方法的总符合率分别为金黄色葡萄球菌92.6%,无乳链球菌91.3%,绿脓杆菌94.6%。该检测方法对奶牛乳房炎的防控、临床监测及疾病诊治具有重要意义。 相似文献
6.
《畜牧兽医科技信息》2019,(12)
黑龙江省某奶牛场奶牛乳房炎发病率高,牛乳中体细胞超标严重,严重影响该奶牛场的经济效益。为了明确引起该奶牛场奶牛乳房炎的主要致病病原菌,筛选敏感药物给出治疗方案。本实验对该奶牛场中30份奶牛乳房炎乳汁进行乳样处理、细菌培养、药敏试验及DNA提取和PCR扩增。通过DNA提取和PCR扩增确定其主要致病病原菌大多为无乳链球菌、停乳链球菌、肠球菌及少量金黄色葡萄球菌。乳样中并未检测出支原体、乳房链球菌及变形杆菌。药敏试验结果表明,该场患病奶牛对阿莫西林、头孢噻呋钠敏感,对一种商品名为惠可宁的硫酸头孢喹肟乳房注入剂耐药性明显。最终得出该厂患乳房炎奶牛的乳汁中分离出的细菌大多数为停乳链球菌,无乳链球菌,肠球菌和少量的金黄色葡萄球菌。当本地区奶牛检测为乳房炎时,可首选阿莫西林,头孢噻呋钠等敏感药物治疗,对奶牛乳房炎的治疗具有积极的指导意义。 相似文献
7.
《畜牧与兽医》2020,(11)
为同时检测大肠杆菌、无乳链球菌和金黄色葡萄球菌,提高对奶牛乳房炎的诊断率,本试验针对3种病原菌的保守序列设计合成了3套特异性引物,通过优化PCR反应体系和反应条件,建立了多重Taq-Man荧光定量PCR检测方法。结果显示,该检测方法具有高特异性,与肠炎沙门菌、肺炎克雷伯菌及鲍曼不动杆菌等无交叉反应;大肠杆菌、金黄色葡萄球菌和无乳链球菌的最低检测浓度分别为10~(3 )、10~2和10~(4 )拷贝数/μL,敏感度较高。DNA各个拷贝数浓度的变异系数均低于2.02%,表明重复性良好。对24份临床采集奶样的检测结果表明,建立的多重Taq-Man荧光定量PCR检测方法可用于临床大肠杆菌、无乳链球菌和金黄色葡萄球菌感染引起的奶牛乳房炎的检测,具有良好的推广应用价值。 相似文献
8.
9.
为建立一种直接从乳样中快速提取细菌DNA的方法,试验通过人工制备8个倍比稀释细菌的乳样,检测了Chelex-100法提取3种奶牛乳房炎主要致病菌(金黄色葡萄球菌、大肠杆菌和无乳链球菌)DNA进行PCR扩增的敏感性,并与苯酚—氯仿法进行了比较分析。结果显示,以Chelex-100法提取乳样中细菌DNA进行PCR扩增具有较高的敏感性,所检测金黄色葡萄球菌、大肠杆菌和无乳链球菌的最小浓度分别为103、102、102 CFU/mL;而使用苯酚—氯仿法提取乳样中各细菌DNA的PCR敏感性均为104 CFU/mL。综上所述,Chelex-100法提取乳样细菌DNA的PCR敏感性可以满足临床检测奶牛乳房炎的需要,显现了简单快速、经济、无污染的优点,为从乳样中直接提取细菌DNA提供了新的思路,对PCR快速检测乳房炎致病菌具有重要意义。 相似文献
10.
大通县城奶牛乳样中葡萄球菌的分离与鉴定 总被引：1，自引：0，他引：1
通过细菌分离培养法对大通县城奶牛乳汁样品36份进行金黄色葡萄球菌的分离，并采用金黄色葡萄球菌常规鉴定方法血浆凝固酶试验、甘露醇发酵试验和接触酶试验进行了鉴定，36份乳样中检出金黄色葡萄球菌阳性7份，阳性率为19．7％(7／36)。 相似文献
11.
12.
13.
魏静 《四川畜牧兽医学院学报》2009,(4):28-32
在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。 相似文献
14.
本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。 相似文献
15.
16.
以国际标准强毒Ｒ株人工感染非免疫产蛋鸡,定时扑杀,分别从鼻窦、眶下孔、气管、肺、气囊、卵巢和输卵管分离ＭＧ,并收集感染鸡所产蛋分离ＭＧ。结果表明,人工感染４８小时后上、下呼吸道及肺已被全面感染,９６小时气囊已被感染,１２０小时输卵管已能分离到ＭＧ,卵巢始终分离不到ＭＧ。人工感染鸡自１４４小时便能在其所产蛋中分离出ＭＧ。药物治疗能在７２小时内消除感染,油乳剂苗则需２４天后逐渐降低蛋内ＭＧ分离率,药物卵内注射、种蛋药浴、高温处理均能杀死卵内ＭＧ,但以研制的种蛋浸泡剂药浴效果为最好。 相似文献
17.
REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air. 相似文献
18.
19.
20.
Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF
Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo. 相似文献