Development of an Immunobinding Assay with Monoclonal Antibodies to Diagnose Mycoplasma bovis in Semen

Veterinary Research Communications -     

Bovine herpesvirus 4-associated postpartum metritis in a Spanish dairy herd

In more than 10 Spanish dairy cows, a bovine herpesvirus 4 (BHV4) associated postpartum metritis was confirmed by virus isolation, BHV4-glycoprotein B (gB) PCR and/or serology. In this study, 12 cows with, and, at the time of sampling, 3 cows without clinical signs of acute postpartum metritis from one large dairy herd in Spain were examined for bacterial and viral infections. Blood, placenta/caruncles and uterine contents were collected between day 1 and day 20 post-calving, and examined for the presence of bacteria and for viruses by virus isolation, BHV4 DNA by BHV4-gB PCR and/or BHV4 antibody titres. Bovine herpesvirus 4 was detected in 83% of the cases with clinical signs of acute postpartum metritis by virus isolation and/or BHV4-gB PCR. An increase of BHV4 antibodies was detected in all examined postpartum metritis cows and in the 3 cows without clinical metritis. Two of these 3 cows developed severe metritis a few dayss after collecting the first blood sample. A concurrent infections of BHV4 and bacteria, mainly Arcanobacterium pyogenes and Streptococcus sp., were detected in 73% of the examined uterine contents collected from postpartum metritis affected cows. This case-report study showed a clear association between BHV4 infections and acute postpartum metritis in dairy cows. In addition, the BHV4-associated postpartum metritis appeared to be an emerging syndrome in this Spanish herd.     

Development of an indirect enzyme-linked immunosorbent assay for detecting equine serum antibodies to the lipopolysaccharide of Salmonella abortusequi

An indirect enzyme-linked immunosorbent assay (IELISA) was developed for the detection of equine serum antibodies to lipopolysaccharide of Salmonella enterica subsp. enterica serovar Abortusequi (LPS), a causative organism of Equine Paratyphoid. The data presented demonstrates that horses immunized with S. abortusequi LPS developed antibodies detectable by the IELISA. By comparison, the tube agglutination test (TAT) did not detect antibody to S. abortusequi LPS as consistently as the IELISA. The data suggests that the IELISA may be a more suitable test for the detection of serum antibodies to S. abortusequi than the TAT.     

太湖地区稻麦二熟制下长期秸秆还田对土壤酶活性的影响

摘要：研究了长期秸秆还田情况下太湖水稻土中与C、N、P、S养分循环及微生物活动有关的荧光素二乙酸酯（FDA）水解酶、酸性磷酸酶、碱性磷酸酶、芳基硫酸酯酶、β-葡糖苷酶、脲酶和脱氢酶等7种酶的活性。结果表明：秸秆还田与无机肥处理都明显增强了所有酶的活性,与对照无肥区的差异达到了显著水平。酶活性对化肥或秸秆还田的敏感度为：β-葡糖苷酶〉脱氢酶、脲酶〉FDA水解酶、碱性磷酸酶〉芳基硫酸酯酶〉酸性磷酸酶。各处理间酶活性顺序为：NPKS2（NPK＋稻秆4500kg/hm^2）〉NPKSI（NPK＋稻秆2250kg/hm^2）〉NPK（N120kg/hm^2,P75kg／hm^2,K150kg/hm^2）〉CK。     

Development of a rapid biological assay for determination of potency of Newcastle disease vaccine (strain I-2)

A rapid biological assay based on incubation time has been developed for determination of the potency of Newcastle disease virus strain I-2 vaccine. It is based on the observation that the interval between inoculation and the first detection of haemagglutinin (HA) depends on the titre of the vaccine inoculated. Chicken embryonated eggs were inoculated with different titres (10⁹, 10⁶ and 10³ EID₅₀/0.1 ml) of vaccine and incubated for 24 h. At hourly intervals, 5 eggs from each vaccine titre were tested for the presence of HA. The results showed that the HA activity was detected from 5, 11 and 15 h after inoculation with vaccine doses of 10⁹, 10⁶ and 10³ EID₅₀, respectively. On the basis of these results it is suggested that if there is no HA detected from 5 to 11 h after inoculation of eggs with the vaccine virus, the vaccine should not be used to vaccinate chickens as it might have an infectivity titre of less than 10⁶ EID₅₀/0.1 ml, which is equivalent to the recommended single chicken dose. It is concluded that measuring the time between inoculation of the vaccine virus and the onset of HA activity might provide an estimate of the titre of the vaccine within 24 h.     

高温胁迫下高羊茅草坪草生理特性研究

从美国引进的39个高羊茅草坪草种分成了5个生态类型,在1999—2000年从每类中找出一个具有典型性状的草种,测定其周年草坪坪质表现、根系活力、叶片可溶性糖含量变化,同时测定各类草种在高温胁迫下保护酶活性与质膜透性的变化。结果表明,根系活力及可溶性糖含量在一定程度上影响着草坪坪质的周年变化。同时,草坪草叶片MDA含量的增长率和草坪草叶片的质膜透性增长率具有显著的正相关效应。在40℃高温胁迫下,草坪草叶片SOD与POD相对活性和MDA含量的增长率及PMP增长率具有显著的负相关效应,5个草种对高温的抗性能力大小顺序是:Gooden>Regi ment>Tomahawk>SR-8210>Brandy。在此基础上,最后对草坪草抗性的准确快速检测方法进行了讨论。     

农产品中基质对酶联免疫分析法检测三唑磷残留的干扰及消除研究

基质干扰是免疫法检测食品中农药残留时经常碰到的问题。以胡萝卜、菠菜、香蕉、梨、稻米和花生等为试材,研究了不同农产品基质对酶联免疫分析法(ELISA)检测三唑磷的影响。结果表明:不同农产品基质对ELISA检测的干扰机制不同,香蕉主要是抑制了辣根过氧化物酶的活性,菠菜、梨和稻米则主要是干扰了抗原和抗体的结合反应,胡萝卜和花生不仅抑制酶的活性,同时还干扰了抗原和抗体的结合反应。将这些农产品的磷酸盐缓冲液(PBS)(含2.5%甲醇)提取液用含有质量分数为0.3%的脱脂奶粉、0.1%的乙二胺四乙酸(EDTA)和含2.5%甲醇的PBS稀释10倍,在12000r/min下离心10min,即可消除其对ELISA检测的干扰。研究结果表明, ELISA法是可用于农产品中三唑磷残留检测的一种简单、快速、有效的方法。     

棉花腺体形成过程中抗氧化酶活性研究

[目的]研究棉花腺体形成过程中相关抗氧化酶的活性,深入了解该时期棉花的生理变化,为培育优良棉花品种提供科学依据。[方法]通过对3个棉花品种(川棉2802、湘棉18、显无N5)腺体形成时期棉酚含量及抗氧化酶活性的测定,研究了棉花的棉酚代谢及腺体的形成。[结果]腺体形成后,川棉2802在萌发初期其棉酚含量逐渐降低,萌发5 d后其棉酚含量缓慢升高;而湘棉18在种子萌发时期,其棉酚含量呈缓慢增长趋势,并逐渐接近川棉2802的棉酚含量。该结果表明,棉花在种子萌发时通过提高SOD、POD和CAT活性,去除超氧自由基,降低过氧化水平,来减小对膜的损害,增强机体抗脂质氧化能力,增加抗逆境能力。[结论]该研究为了解棉花腺体形成时期的特点及利用腺体性状的转育奠定了科学基础。     

低温胁迫下Ca2＋-CaM信使系统对三尖杉幼苗保护酶活性的影响

用一定浓度的CaCl2、LaCl3、EGTA、CPZ等4种效应剂根施三尖杉1年生幼苗,在25℃下预处理10d后,移入-3℃下进行低温胁迫处理,分别在胁迫后24、48、72、96h时测定过氧化氢酶（CAT）、过氧化物酶（POD）和超氧化酶（SOD）等3种保护酶的活性。结果表明：根施外源CaCl2结合低温处理可增强三尖杉幼苗3种保护酶的活性,缓解低温处理过程中保护酶活性的下降;但三尖杉幼苗内源保护酶活性可被EGTA、LaCl3以及CPZ等3种效应剂抑制。说明三尖杉幼苗抗冻性的形成受Ca2＋-CaM信号传导途径的调控。     

玉米秸秆腐解对石灰性褐土酶活性和有机质质量分数动态变化的影响

通过室内培养,研究玉米秸秆腐解量与腐解时间的关系,以及不同秸秆添加量对石灰性褐土酶活性和有机质质量分数动态变化的影响。结果表明,玉米秸秆的腐解量随培养时间延长而增加,在培养的第45天,腐解量可达45%左右,而45天后腐解速度明显下降。在腐解的前60d,常规秸秆添加处理的腐解速率小于加倍量秸秆添加量处理,而在培养后期处理间腐解速率无差异,且腐解速率均随腐解时间延长而减小。土壤过氧化氢酶活性随培养时间的延长呈先增加后降低再增加的趋势,但不同秸秆量处理土壤过氧化氢酶活性差异不显著;碱性磷酸酶活性随秸秆添加量增加而增大,且均随培养时间延长而提高。土壤有机质和活性有机质质量分数均随培养时间延长而增加,且随秸秆添加量的增加而显著增加。