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从山羊瘤胃内容物中提取瘤胃细菌总DNA,以瘤胃中两种主要纤维分解菌为目的菌种,分别依据其16S rDNA序列设计引物,利用PCR技术对其16S rDNA 进行体外扩增,对扩增产物测序后进行同源性分析.结果表明:以提取的瘤胃细菌总DNA为模板,采用PCR技术可成功地从山羊瘤胃内容物中扩增出黄色瘤胃球菌和产琥珀酸丝状杆菌16S rDNA的特异性片段;测序后与GenBank中的序列进行比对,表明黄色瘤胃球菌与原序列(AF104841)的同源性达到99.4%,产琥珀酸丝状杆菌与原序列(AJ505937)的同源性达到94.6%. 相似文献
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利用实时定量PCR研究不同蛋白质饲料对嗜淀粉瘤胃杆菌生长参数的影响 总被引：1，自引：0，他引：1
本试验主要研究不同蛋白质饲料对嗜淀粉瘤胃杆菌(Ruminobacter amylophilus)生长的影响.选用4只装有瘤胃瘘管的徐淮白山羊羯羊,分别饲喂以羽毛粉、玉米蛋白粉、豆粕和鱼粉为蛋白质补充料的4组日粮,进行4×4拉丁方设计的试验,并采用实时定量PCR(RT-PCR)技术对嗜淀粉瘤胃杆菌进行定量分析.结果表明,豆粕组瘤胃氨氮浓度最高,并显著高于其他3组(P<0.05);细菌总数在组间差异不显著(P>0.05);嗜淀粉瘤胃杆菌的密度及其数量在细菌总数中的比例均以豆粕组最高,且组间差异显著(P<0.05).总之,豆粕饲料适宜于嗜淀粉瘤胃杆菌的生长,RT-PCR技术适用于瘤胃微生物的定量研究. 相似文献
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旨在研究山羊瘤胃产甲烷古菌的多样性并与其他动物瘤胃进行比较。采用产甲烷菌特异性引物Met86F/Met1340R研究山羊瘤胃产甲烷古菌16SrRNA基因的多样性,随机挑选100个克隆进行序列分析。结果表明,所有克隆经限制性内切酶酶切后共获得16个OTU,绝大多数为甲烷短杆菌,其中与甲烷短杆菌菌株AK-87、ZA-10、OCP、SM9和30Y序列最相近的克隆数分别为58%、19%、7%、2%和2%,还有与Methanosphaera stadtmanae(1%)、Methanobacterium aarhusense(2%)、Aciduliprofundum boonei(3%)、Methanobrevibacter sp.AK-87(4%)和Methanobrevibactersp.1Y(2%)等相似的古菌序列。将研究结果与已经报道的关于牛和绵羊瘤胃产甲烷菌多样性比较,发现不同PCR引物可检测出不同菌群结构,而饲料类型、动物种类可影响瘤胃产甲烷菌的菌群结构。 相似文献
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体外培养条件下氨基酸对瘤胃蛋白质降解菌生长限制性的影响 总被引：1，自引：0，他引：1
本试验旨在研究体外培养条件下氨基酸对瘤胃蛋白质降解菌[嗜淀粉瘤胃杆菌(R.amylophilus)、溶纤维丁酸弧菌(B.fibrisolvens)和柄瘤胃普雷沃氏菌(P.ruminicola)]生长限制性的影响.以4头安装永久性瘤胃瘘管的徐淮山羊为瘤胃液供体,采用人工模拟山羊瘤胃发酵装置进行培养.通过特定氨基酸缺省的方法,设定5个组:全量氨基酸组、蛋氨酸缺省组、赖氨酸缺省组、支链氨基酸缺省组以及芳香族氨基酸缺省组.结果表明:特定氨基酸缺省对嗜淀粉瘤胃杆菌、溶纤维丁酸弧菌和栖瘤胃普雷沃氏菌的生长限制性作用有所差异.特定氨基酸的缺省对嗜淀粉瘤胃杆菌、溶纤维丁酸弧菌和栖瘤胃普雷沃氏蒲的生长限制性作用(相对于全量氨基酸组)排序分别为芳香族氨基酸>赖氨酸>支链氨基酸>蛋氨酸;赖氨酸>芳香族氨基酸>支链氨基酸>蛋氮酸;支链氨基酸>芳香族氨基酸>蛋氨酸>赖氨酸.由此得出,在体外培养条件下,氨基酸缺省将影响瘤胃蛋白质降解蔚的生长. 相似文献
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旨在建立一种有效的定量方法监测肉牛在运输前后瘤胃内3种主要细菌的变化,本试验选用8头体质量相近的健康夏南牛,在运输前、运输中6h、运输后1、3、7、15d时间段采集瘤胃液。分别以黄化瘤胃球菌、嗜淀粉瘤胃杆菌、脂解厌氧弧杆菌的16SrRNA序列设计引物,运用Real-time PCR技术研究运输应激对肉牛以上3种瘤胃微生物的影响。结果显示,与运输前相比,黄化瘤胃球菌在运输中6h、运后1d浓度显著升高（P〈0.05）。嗜淀粉瘤胃杆菌在运后3d浓度升高到运输前的13.86倍,随后逐渐降低,15d时为运输前8.42倍。脂解厌氧弧杆菌在运输后3d数量出现极显著的降低（P〈0.05）,在7、15d逐渐恢复。结果表明,运输应激对黄化瘤胃球菌影响不大;脂解厌氧弧杆菌在运输应激后出现显著的降低,运输后半月内逐渐恢复正常水平。嗜淀粉瘤胃杆菌则呈现明显的上升趋势。采用Real-time PCR方法对瘤胃细菌进行定量可以快速有效的反映出运输应激对菌的数量变化趋势。 相似文献
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低淀粉饲粮条件下不同瘤胃降解淀粉水平对体外瘤胃发酵的影响 总被引：1，自引：0，他引：1
本试验旨在研究玉米作为淀粉来源的低淀粉饲粮条件下不同瘤胃降解淀粉(RDS)水平对体外瘤胃发酵的影响。以3头安装有永久性瘤胃瘘管的健康荷斯坦奶牛作为瘤胃液供体,各组分别以不同RDS水平饲粮作为发酵底物,体外产气法测定培养48 h时产气量和瘤胃发酵参数以及24 h时瘤胃微生物区系变化。结果表明:1)随着饲粮RDS水平的提高,体外培养48 h时产气量、潜在产气部分和产气速率呈线性升高(P0.05),快速发酵部分产气量呈线性下降(P0.05),干物质消失率呈线性升高(P0.05);2)随着饲粮RDS水平的提高,体外培养48 h时发酵液微生物蛋白、乙酸、丙酸、丁酸和总挥发性脂肪酸浓度呈线性升高(P0.05),pH和氨态氮浓度没有显著变化(P0.05);3)随着饲粮RDS水平的提高,体外培养24 h时发酵液中白色瘤胃球菌和嗜淀粉瘤胃杆菌的相对数量呈线性升高(P0.05),黄色瘤胃球菌、琥珀酸丝状杆菌、溶纤维丁酸弧菌、牛链球菌和溶淀粉琥珀酸单胞菌的相对数量没有显著变化(P0.05)。综合考虑,低淀粉饲粮条件下提高RDS水平有利于瘤胃发酵。 相似文献
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实时荧光定量PCR对瘤胃纤维分解菌定量方法的构建 总被引：2，自引：1，他引：1
试验构建了白色瘤胃球菌、黄色瘤胃球菌及产琥珀酸丝状杆菌等3种瘤胃纤维分解菌实时荧光绝对定量PCR的标准品及标准曲线,以用于纤维分解菌的定量测定。提取瘤胃微生物总DNA,以各菌特异性引物进行PCR扩增,回收纯化PCR产物,与pMD18-T Vector连接并转化到大肠杆菌。用氨苄青霉素培养基筛选阳性重组质粒,提取含目的片段质粒DNA,通过PCR及测序鉴定重组质粒。根据OD值确定浓度,将梯度稀释的质粒作为模板,进行荧光定量PCR反应做出标准曲线。结果表明:所构建的3条标准曲线具有很高的相关性(R2>0.999),并获得了高扩增效率产物(白色瘤胃球菌为101%、黄色瘤胃球菌为98.0%、产琥珀酸丝状杆菌为97.7%),说明成功构建了瘤胃纤维分解菌实时绝对定量PCR的标准品和标准曲线,为定量研究瘤胃纤维分解菌奠定基础。 相似文献
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旨在观察不同NFC/NDF比日粮对奶山羊瘤胃细菌及瘤胃和血浆中内毒素及组胺含量的影响。选用6只安装有永久性瘤胃瘘管的泌乳期关中奶山羊为试验动物,试验分四期进行,每期10d,依次饲喂NFC/NDF比分别为1.02(Ⅰ期)、1.24(Ⅱ期)、1.63(Ⅲ期)、2.58(Ⅳ期)的4种日粮,以逐渐增加日粮精料的方式诱导奶山羊发生亚急性瘤胃酸中毒(SARA)。分别利用pH动态监测记录系统和滚管培养法测定瘤胃pH、瘤胃细菌总数、淀粉分解菌及坏死梭形杆菌的浓度;以鲎试验试剂法和荧光分光光度法测定内毒素和组胺含量。结果表明:随着日粮NFC/NDF比的增大,瘤胃pH显著降低(P0.05),并且瘤胃pH下降速率和下降幅度也随之加快;日粮NFC/NDF比为2.58时,瘤胃内组胺和内毒素的浓度明显增加(P0.05),坏死梭形杆菌和淀粉分解菌的数量也显著升高(P0.01)。内毒素、组胺和坏死梭形杆菌三者浓度的异常增加对SARA发生和发展起重要作用。 相似文献
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运用Real-time PCR方法研究日粮添加豆油与胡麻油对肉牛瘤胃纤维分解菌数量的影响 总被引：3，自引：0，他引：3
本研究分别以产琥珀酸丝状杆菌(Fibrobacter succinogenes)、黄色瘤胃球菌(Ruminococcus flavefaciens)、白色瘤胃球菌(Ruminobacter albus)和溶纤维丁酸弧菌(Butyrivibrio fibrisolvens)16S rDNA序列设计引物,运用Real-time PCR技术研究日粮中添加豆油与胡麻油对肉牛上述4种瘤胃纤维分解菌数量的影响.结果表明,与对照组(CK)相比,添加豆油组(LOC1)和胡麻油(LOC2)组,产琥珀酸丝状杆菌(Fibrobacter succinogenes)、黄色瘤胃球菌(Ruminococcus flavefaciens)、白色瘤胃球菌(Ruminobacter albus)和溶纤维丁酸弧菌(Butyrivibrio fibrisol-vens)数量显著减少(P<0.05),分别降低了78%和31%、30%和36%、27%和23%、6%和13%.通过该方法的结果表明日粮中添加4%的豆油和胡麻油显著减少了瘤胃中纤维分解菌,对产琥珀酸丝状杆菌的影响明显.而且采用Real-time PCR方法对瘤胃纤维分解菌进行定量,可以快速有效反映出在日粮改变的情况下菌的数量变化趋势,相对于传统计数方法更直观、快捷与准确. 相似文献
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焦万洪 《养殖与饲料．饲料世界》2014,(12):37-37
1饲料蛋白质被瘤胃微生物降解瘤胃中有很多细菌能分泌蛋白酶,具有降解蛋白质的能力。最熟知的有嗜淀粉拟杆菌、溶纤维丁酸弧菌和栖瘤胃普雷沃氏菌这3种细菌。瘤胃真菌也具有分解蛋白质的能力。瘤胃厌氧性真菌菌株都可产生金属蛋白酶,N.frontalis可产生相当于细菌产生的高活性蛋白酶。 相似文献
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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
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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
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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
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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
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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