荧光定量RT-PCR检测H5亚型禽流感病毒方法的建立与验证

根据H5亚型禽流感病毒HA基因上的保守序列,设计合成引物,以H5亚型禽流感病毒HA基因重组质粒为标准品绘制标准曲线,建立了荧光定量逆转录聚合酶链反应检测方法。结果表明,本试验建立的标准曲线循环阈值(Ct值)与模板浓度具有良好的线性关系,相关系数为0.995,灵敏度约为8拷贝/μL,相当于8个AIV颗粒,对新城疫病毒和其他禽病病毒无交叉反应,特异性好、重复性佳,为H5亚型禽流感病毒检测提供了一种特异、敏感、快速、低成本、高通量、生物安全性好的定量检测方法。对178份临床泄殖腔棉拭子样品的检测,该方法结果与经典病毒分离方法符合率大于90.0%,在禽流感病毒临床样品快速筛检、流行病学监测等方面显示了良好的应用前景。     

中国部分山羊品种线粒体DNA D-loop序列遗传多样性分析

分析了中国部分本地山羊品种(18个品种200个体)以及在中国饲养的引入品种(4个品种25个体)线粒体DNA控制区(D-loop)的全序列,结合已报道的世界其它地方的山羊(15个品种77个体)以及2只野山羊的线粒体DNA控制区(D-loop)全序列共304个体,研究结果表明:山羊线粒体DNA控制区约为1 212 bp,检测到228个变异位点,203种单倍型,单倍型多样度为0.993±0.001;核苷酸多样度0.018±0.001。中国部分山羊的变异类型主要为类型A和B,而在巴基斯坦山羊中还检测到类型C和D。比较分析结果表明中国山羊遗传多样性和基因交流比中国黄牛品种要高。     

高效液相色谱法测定猪血浆及组织中喹赛多及其脱二氧代谢物

建立了测定猪血浆及肝脏、肾脏、肌肉等组织中的喹赛多及其代谢物脱二氧喹赛多的高效液相色谱(HPLC)法.血浆样用甲醇沉淀蛋白后离心取上清液进样测定.组织样先用乙腈匀浆,用正己烷脱脂后作HPLC检测.色谱柱为ODS C18柱;流动相血浆样测定为乙腈水(2080),组织样测定为甲醇水(4258),流速为1.0 mL/min;紫外检测波长305 nm.药物工作液质量浓度范围为0.005～0.500 mg/L时,血浆样品及组织样品测定条件下药物质量浓度与响应值均具良好线性关系,相关系数＞0.999.血浆中药物质量浓度为0.02、0.10、0.50 mg/L时,喹赛多及脱二氧喹赛多的回收率均大于70%,组织中药物含量为0.05、0.20、1.00μg/g时,肌肉样品的回收率均大于70%,肝脏、肾脏样品则为50%～80%.本试验条件下,喹赛多及其脱二氧代谢物的最低检出质量浓度,血浆样品分别为0.01、0.02mg/L,组织样品2种检测物均为0.025μg/g.测定了工作液3种质量浓度0.01、0.05、0.25 mg/L的仪器精密度,日内相对偏差＜8.0%,日间相对偏差＜17.0%.     

去毒桐粕及其在畜牧生产上的应用

油桐(V ernicia fordii(H em sl)A iry-Shaw)属于大戟科(E uphorbiaceae)油桐属(V ernicia Lour),为中亚热带落叶乔木,原产于我国,有着悠久的栽培历史,是著名的经济植物之一。其主要产品是桐油,为最佳干性油之一,在工业上具有广泛用途。据统计,目前在国民经济领域中,直接或间接使用桐油的产品达l000种以上,且随着各种工业的日益发展,桐油的用途也将更为广泛。桐饼(粕)主要是油桐籽经榨取桐油后所剩余的副产品,由于不同的制油方法以及油脂厂所收购的桐果品种不一,且加工过程中加入的桐壳比例不同,因此在桐籽制取桐油的过程中,由机械压榨法…     

Characterization of culture supernatant proteins from Brucella abortus and its protection effects against murine brucellosis

In this study, we characterized the secreted proteins of Brucella abortus into the enriched media under the bacterial laboratory growth condition and investigated the pathogenic importance of culture supernatant (CS) proteins to B. abortus infection. CS proteins from stationary phase were concentrated and analyzed using 2D electrophoresis. In MALDI TOF/TOF analysis, more than 27 proteins including CuZn SOD, Dps, Tat, OMPs, Adh, LivF, Tuf, SucC, GroEL and DnaK were identified. Cytotoxic effects of CS proteins were found to increase in a dose-dependent manner in RAW 264.7 cells. Upon B. abortus challenge into phagocytes, however, CS proteins pre-treated cells exhibited lower bacterial uptake and intracellular replication compared to untreated cells. Immunization with CS proteins induced a strong humoral and cell mediated immune responses and exhibited significant higher degree of protection against virulence of B. abortus infection compared to mice immunized with Brucella broth protein (BBP). Taken together, these results indicate that B. abortus secreted a number of soluble immunogenic proteins under laboratory culture condition, which can promote antibody production resulted in enhancing host defense against to subsequently bacterial infection. Moreover, further analysis of CS proteins may help to understand the pathogenic mechanism of B. abortus infection and host–pathogen interaction.     

Correlation Analysis on the Fineness and Single Fiber Strength of Alpaca Wool Fiber

In this study,321 samples were tested to evaluate the correlation between fineness and single fiber strength of alpaca fiber. The results showed that fineness average of lateral part was 24.30 μm,subject range was from 18.01 to 27.00 μm,accounted for 72% of the total samples;The strength average was 8.31 cN,subject range was from 5.01 to 11.00 cN, accounted for 88% of the total samples, it showed that the overall fineness of alpaca wool fiber was fine,the strength could satisfy the textile technology. The correlation results showed that the fineness of different parts (neck, shoulder, thigh, abdomen, lateral and back) in different groups (sex, age) all reflected that the wool fiber of the back was the finest,the strength was the minimal;And that of abdomen was the thickest,the strength was the maximum. On the other hand, we knew the greater the strength, the fineness was thicker; The smaller the strength, the fineness was finer from the distribution of different fineness range corresponded to the strength. In addition, there was also a positive correlation between them in differernt groups (sex,age),the fineness of adult alpaca was thicker than yearling alpaca, the male alpaca was thicker than female alpaca; The strength of adult alpaca was stronger than yearling alpaca, the male alpaca was stronger than female alpaca. The above results showed that the fineness of alpaca wool fiber was positively correlated with the single fiber strength.     

Effect of topical anesthesia on evaluation of corneal sensitivity and intraocular pressure in rats and dogs

Objective To determine the effect of 0.5% proparacaine in tonometry by evaluating corneal touch threshold (CTT) and intraocular pressure (IOP). Animal studied Nine rats (18 eyes, Sprague–Dawley) and 10 dogs (20 eyes, Beagle) Procedures The IOP and CTT were measured in each eye before and after topical anesthesia with 0.5% proparacaine. The IOP was evaluated using Tonopen for dogs and Tonolab for rats. The corneal sensitivity was evaluated by CTT through a Cochet–Bonnet aesthesiometer. Results The mean IOP was not significantly changed in rats or dogs before and after topical anesthesia. However, after application of proparacaine, CTT was significantly increased in both animal groups compared with that before application of proparacaine. Conclusion From this study, topical anesthesia was found to significantly lower the corneal sensitivity but have little effect on IOP measurements. In ophthalmologic examination, topical anesthesia can be used to reduce corneal sensation without an effect on IOP.     

柴黄颗粒对人工感染鸡传染性支气管炎的防治研究

为探讨柴黄颗粒对鸡传染性支气管炎的防治效果,将400只15日龄海兰褐鸡随机均分8组:柴黄颗粒预防组,低、中、高剂量治疗组,中、西药对照组,模型对照组和健康对照组.除健康对照组外,其余各组均接种鸡传染性支气管炎病毒M41,预防组于攻毒前7d饮水中加入柴黄颗粒进行预防,各药物治疗组于攻毒48 h后进行药物治疗,直到临床症状消失.结果表明,感染前7d给予2 g/L柴黄颗粒对人工感染鸡传染性支气管炎有较好的保护作用,使其发病率降低至50％,与模型对照组比较差异显著(P＜0.05);4 g/L柴黄颗粒治疗组死亡率为5％,与模型对照组比较差异极显著(P＜0.01),治愈率为92％,与西药对照组差异极显著(P＜0.01),效果优于中药对照组.柴黄颗粒中剂量组攻毒第7天抗体滴度达到1 531.33±389.91,与模型对照组差异极显著(P＜0.01),并随时间增加而增加.2g/L预防剂量和4 g/L治疗剂量的柴黄颗粒对鸡呼吸型传染性支气管炎有较好防治效果.     

植酸酶包埋后酶活和热稳定性变化的规律研究

为了了解植酸酶在包埋状态下酶活及酶性的变化,分别对酶的释放情况和热稳定性进行了研究.结果表明,随着时间的延长,包埋酶释放出的酶活逐渐上升(P＜0.01),60min时达到最大值;而未包埋酶释放出的酶活逐渐下降(P＜0.01),60 min时达到最小值.包埋酶和未包埋酶在30、40、50、60、70、80、90和100℃的条件下,随着温度的升高,残留的植酸酶活力下降(P＜0.01).对于包埋酶,当温度升高到70℃时,植酸酶相对酶活仍保持在93%(P＞0.05);当温度升高到80℃时,植酸酶的相对酶活才明显下降(P＜0.01),不过在100℃时,仍能保持67%的酶活(P＜0.01).对于未包埋酶,当温度升高到60℃时,植酸酶相对酶活明显下降(P＜0.01);当温度升高到100℃时,植酸酶相对酶活仅有23%(P＜0.01).通过在统一温度条件下,对包埋酶和未包埋酶处理组的相对酶活的对比发现,在50℃以下时,二者差异不显著,当温度超过60℃时,包埋酶的相对酶活极显著高于未包埋酶.     

猪瘟疫苗的豚鼠变态反应试验

本研究用猪瘟疫苗初次致敏豚鼠未引起异常反应,两周后进行第2次激发可见明显的异常反应,甚至死亡,这证明了猪瘟疫苗可能引起变态反应的发生。为了找到疫苗中的真正过敏原,本研究进行了疫苗中主要成分的分组试验,结果表明犊牛血清的致敏作用最强,其次为猪瘟病毒,牛睾丸细胞最弱。同时进行了4种致敏和激发途经的比较实验,并用ELISA法测定了致敏和激发后血清中的IgE水平。结果表明各种致敏途径均可致敏,但激发时,以静脉注射最为严重,且致敏后和激发后均未检测到血清中的IgE水平有显著提高。