过氧化氢溶液对鸡新城疫病毒的杀灭效果试验

为考察过氧化氢溶液对鸡新城疫病毒的杀灭效果,依据中华人民共和国农业部《兽用消毒剂鉴定技术规范》(试行)(1992年版),以鸡胚感染法进行了不同稀释倍数(5、10、20倍稀释)的过氧化氢溶液对新城疫病毒的定量、定性杀灭效果。定量试验结果显示,10倍稀释的过氧化氢溶液与鸡新城疫病毒作用5 min,病毒灭活率为100%,定性试验结果显示,20倍稀释的过氧化氢溶液与鸡新城疫病毒作用60 min,对鸡新城疫病毒的灭活率在80%以上。表明过氧化氢溶液与鸡新城疫病毒作用5min,有效浓度为10倍稀释,其最低有效浓度为20倍稀释,该浓度对新城疫病毒的最快有效时间为60 min。     

戊二醛苯扎溴铵溶液对载体的消毒试验

采用载体浸泡定量杀菌试验,考察戊二醛苯扎溴铵溶液对载体片染菌的杀灭效果。参考中华人民共和国农业部《兽用消毒剂技术规范》(1992年版),观察不同稀释倍数(800、1600、6.25、12.5)在不同作用时间(5、30 min)其对载体片染菌的杀灭效果。戊二醛苯扎溴铵溶液800倍稀释,与载体片上的细菌作用5 min,以及6.25倍稀释,与蜡样芽孢杆菌8008作用30 min,杀菌率均为100%。说明戊二醛苯扎溴铵溶液有效消毒浓度为800倍稀释,灭菌浓度为6.25倍稀释。     

液体植酸酶的稳定性研究

此文研究了液体植酸酶在不同贮存温度下的酶活稳定性,以及不同稀释倍数和调节稀释后酶液的pH时液体棱酸酶的活性和物理性状的影响,同时探讨了液体植酸酶与液体木聚糖酶复配的稳定性能。试验结果表明：液体植酸酶在低温下贮存酶活稳定性较好;不同稀释倍数对液体植酸酶活性无明显影响,但稀释倍数较大显著影响？植酸酶溶液的物理性状;通过调节稀释后的植酸酶溶液的pH可以显著改善其物理性状。此外,与液体木聚糖酶复配无论是复配后酶液蛇活性还是其物理性状均具有很好盼稳定性。     

聚丙烯酰胺凝胶电泳快速鉴定牛乳蛋白质的研究

试验利用SDS-聚丙烯酰胺凝胶电泳技术对牛乳进行快速质量检测研究,通过对样品稀释倍数、电压条件、染色时间、漂洗时间及次数等条件进行摸索,获得了条带清晰的牛乳蛋白电泳图谱.结果表明,快速聚丙烯酰胺凝胶电泳法鉴定牛乳蛋白质的最佳条件是,用8%分离胶在120V电压下电泳.使用超级灵敏度蛋白质电泳快速染液染色,煮沸染色1 min,振荡10min,可实现快速鉴定牛乳蛋白的目的.     

犬细小病毒ELISA抗体检测试剂盒的研制及应用

本研究以纯化的犬细小病毒(CPV)重组VP2蛋白为包被抗原,建立了检测CPV抗体的间接ELISA 方法,其抗原包被浓度为0.158μg/mL;待检血清稀释倍数为1:100,作用时间为60 min;羊抗犬酶标抗体稀释倍数为1:2 000,作用时间为60 min;底物作用时间为10 min;判定标准为S/P≥O.282时为阳性,S/P≤0.226时为阴性.该方法与犬瘟热、犬传染性肝炎、犬副粘病毒病、犬波特氏杆茵痛等4种犬常见传染病阳性血清无交叉反应,具有较好的特异性;批内、批间重复试验变异系数均小于15%,显示该方法具有良好的重复性;与商品化CPV ELISA试剂盒进行比较,符合率为96.5%.本研究为现地免疫犬群抗体监测和进行CPV流行病学调查提供了一种简便的血清学诊断方法.     

鼻气管鸟杆菌间接ELISA检测方法的建立及应用

提取A型鼻气管鸟杆菌(ORT)外膜蛋白,经纯化后包被反应板,通过方阵滴定确定最佳抗原包被浓度、血清稀释倍数、兔抗鸡酶标二抗稀释倍数,建立了检测ORT抗体的间接ELISA方法.经方阵滴定确定最佳抗原包被质量浓度为11.85 mg/L,血清样品稀释倍数为1∶20,兔抗鸡IgG辣根过氧化物酶结合物最佳稀释倍数为1∶1 000,抗原抗体最佳结合时间为1 h,血清与二抗最适反应时间为45 min.试验结果确定的判定标准为:样品D490 nm≥0.639判定为阳性,D490 nm≤0.350判定为阴性,介于两者之间的为可疑.试验证明该方法具有良好的稳定性、重复性和特异性,将205份疑似ORT的鸡血清用本方法进行检测,其阳性检出率为11.7%(24/205).     

戊二醛苯扎溴铵溶液对H9N2亚型禽流感病毒的灭活效果分析

采用悬浮杀灭试验,考察戊二醛苯扎溴铵溶液对H9N2亚型禽流感病毒的灭活效果。将戊二醛苯扎溴铵溶液通过不同稀释倍数(1 250、2 500、5 000、10 000、20 000)、在不同作用时间(5、10 min)观察其对H9N2亚型禽流感病毒的灭活效果。结果显示,戊二醛苯扎溴铵溶液在10000倍稀释,与H9N2亚型禽流感病毒作用5 min或10 min,灭活率达99.90%以上;在20 000倍稀释,作用5 min或10 min,灭活率均小于99.90%。表明戊二醛苯扎溴铵溶液在10 000倍稀释与H9N2亚型禽流感病毒(20±1)℃水浴作用5 min,即可对H9N2亚型禽流感病毒达到很好的灭活效果。     

饲用中性蛋白酶酶活测定的影响因子的研究

本文介绍了福林酚法测定中性蛋白酶的原理、步骤,并重点研究了福林法测定中性蛋白酶在实际应用中的一些问题。研究表明,无论何种来源的固体蛋白酶都可以用0.3mol/L氯化钠作为浸提剂,浸提时间为30min;酶促反应的时间为5min,显色反应为20min;酶液合适的稀释倍数为最后测定酶活时OD值不超过0.5;不同厂家生产的酪素底物对测定结果有影响,底物浓度应保持在1%;酶促反应的温度和pH分别为40℃和7.0。     

戊二醛苯扎溴铵溶液对鸡新城疫病毒的杀灭效果研究

考察戊二醛苯扎溴铵溶液消毒剂对新城疫病毒的杀灭效果。参考中华人民共和国农业部《兽用消毒剂技术规范》(1992年版),采用悬浮杀灭试验,以鸡胚感染法,将其通过不同稀释倍数(800、1600、3 200)、在不同时间段(5、10、15、30、60 min)观察其对新城疫病毒的杀灭效果。结果:戊二醛苯扎溴铵溶液在20℃水浴中,1 600倍稀释,作用5min,对鸡新城疫病毒的灭活率在80%以上。结论:戊二醛苯扎溴铵溶液在20℃水浴中,对鸡新城疫病毒的最低有效浓度为1 600倍稀释,该浓度对新城疫病毒的最快有效时间为5 min。     

猪沙波病毒抗体间接ELISA检测方法的建立及初步应用

以原核表达重组蛋白作为抗原建立了猪沙波病毒(SaV)抗体的间接ELISA检测方法。使用矩阵滴定法确定抗原包被浓度为6.4μg/mL,血清稀释倍数为1∶50。试验选用1%BSA作为封闭液,血清和二抗的最佳作用时间分别为60 min和40 min。选择无致癌的TMB为底物,确定TMB最佳反应时间为10 min。通过对20份猪阴性血清样品的检测,计算阳性判定值为0.203。通过交叉试验、批内和批间重复性试验证明,本研究建立的SaV间接ELISA检测方法具有特异性高,重复性好的特点,可用于猪沙波病毒抗体的检测。     

Comparative effects of the human protein C activator, Protac, on the activated partial thromboplastin clotting times of plasmas, with special reference to the dog.

The commercial snake venom extract, Protac, is a specific activator of the anticoagulant zymogen, protein C (PC) in human plasma. This specific action has led to its use in developing coagulation-based and amidolytic-based assays for the diagnosis of quantitative and/or qualitative PC deficiency states in human beings. The purpose of the present study was to compare the effects of Protac on the activated partial thromboplastin times (APTT) of human, bovine, equine, and canine plasmas in order to determine the potential value of this venom extract as an activator in functional PC assays in these domestic animal species. As expected, Protac significantly prolonged the APTT of normal human plasma, but had no effect on plasma known to be devoid of PC. Clotting times were prolonged by 34%-214% with concentrations of venom activator ranging from 0.1-1.0 U/mL. Under identical conditions, Protac prolonged the APTT of equine plasma by 11%-98% over control times. Even more dramatic was the inhibitory effect of Protac on the clotting of bovine plasma, extending the APTT more than 3-fold at a venom concentration of 0.1 U/mL. At higher venom concentrations, most bovine plasmas remained unclotted after 300 s (control time 34.1 s). Under similar conditions, the canine APTT was unaffected by Protac, even when the venom concentration was increased to 3 U/mL. In order to determine the reason for the lack in response of canine plasma, the concentration of the APTT reagent was altered (decreased), exposure time of the plasma to the Protac was increased from 2 min to 9 min, and the plasma was diluted to assess for the potential existence of plasma PC inhibitors. Protac caused an unexpected shortening of the APTT when the contact activator reagent was diluted. Increasing the exposure time had no effect. Although a slight prolongation of the canine APTT was detected when the plasma was diluted, the presence of strong plasma PC inhibition was considered an unlikely cause of the lack of significant anticoagulant action. The failure of Protac to exert a strong inhibitory effect on the canine APTT, as well as to generate amidolytic activity, suggests that this venom extract does not stimulate the production of activated PC activity in canine plasma. This may result from molecular differences in the canine PC molecule that prevent the formation of the stoichiometric complex of venom extract, APTT reagent, and canine protein, a complex thought to be essential for the PC-activating function of Protac. Protac may be suitable as an activator of PC in bovine and equine plasmas; however, it appears ineffective in generating anticoagulant activity in canine plasma.     

过氧乙酸戊二醛复方消毒剂与氨气的互作

为研究氨气与过氧乙酸戊二醛复方消毒剂(简称复方消毒剂)相互中和的效果及氨气对消毒效果的影响,通过测定不同剂量512倍稀释的复方消毒剂,在不同作用时间、不同氨气浓度时,喷雾消毒前后气雾柜内细菌数量和氨气质量浓度,计算杀菌率和氨气中和率,以确定512倍稀释的复方消毒剂对氨气的中和效果及高浓度氨气对其杀菌效果的影响.结果显示...     

Comparison of Different Extenders with Defined Protein Composition for Storage of Stallion Spermatozoa at 5°C

To maintain the fertility of stallion spermatozoa during cooled storage, extender media are added to semen. In this study, three semen extenders were compared: EquiPro which contains defined caseinates and whey proteins instead of dried skim milk. The extender is provided in dry form and dissolved in distilled water prior to use. EquiPro TM has the same composition as EquiPro but is provided in a sterilized ready-to-use liquid form. AndroMed-E contains soybean lecithin as protein source. Semen was collected from seven stallions. Ejaculates were divided into three aliquots, diluted with the different extenders and stored at 5 degrees C for 4 days. Total motility, membrane integrity, average path velocity (VAP), curvilinear-velocity (VCL), straight-line velocity (VSL), distance average path (DAP), distance curved line (DCL) and distance straight line (DSL) were determined by computer-assisted analysis. Total motility decreased in all extenders during storage. The parameters VAP, VCL, VSL, DAP, DCL and DSL in semen diluted in EquiPro TM at most times and in semen diluted in AndroMed-E at some times were lower than in semen diluted in EquiPro (p < 0.05). Viability on days 0 and 4 was lowest in semen diluted in AndroMed-E (p < 0.05). Velocity decreased faster when semen had been diluted in the sterilized liquid extender EquiPro TM or in AndroMed-E compared with the dry formula of EquiPro. Therefore the liquid sterilized EquiPro despite no difference in its chemical composition differs from the dry, non-sterilized EquiPro extender. Heat sterilization apparently changes effects of the extender on spermatozoa.     

表面等离子共振生物传感器检测培氟沙星的方法研究

研究表面等离子共振生物传感器检测缓冲液中培氟沙星的方法,为样品的检测提供依据。采用抑制法,将培氟沙星与卵清蛋白的偶联物固定在芯片上,将抗体与不同浓度的培氟沙星混合后通入芯片表面,优化反应条件,构建标准曲线。结果显示,选择1∶1000稀释的抗体,流速7 μL/min,反应时间4 min,构建标准曲线,可以达到较低的检测限。利用SPR生物传感器建立快速检测缓冲液中培氟沙星的方法,检测限为0.8 ng/mL,检测时间为4 min,为样品检测奠定基础。     

垃圾堆肥放线菌发酵过滤液对高羊茅初期生长的影响

通过向草坪基质中加入不同浓度生活垃圾堆肥放线菌发酵滤液,研究了堆肥放线菌发酵液对草坪草高羊茅萌发及初期生长的影响。结果表明:各浓度放线菌发酵滤液对高羊茅的生长指标和叶绿素含量均有显著的促进作用,其中,以稀释4倍的放线菌发酵滤液的促进效果最为明显。在稀释4倍的放线菌发酵滤液的处理下,高羊茅的发芽率、株高、地上干重、地下干重和叶绿素含量分别比对照提高了17.3%、17.9%、20.0%、66.7%和60.3%。虽然,不同浓度的放线菌发酵滤液对高羊茅的萌发和初期生长均具有一定的促进作用,但以稀释4倍的放线菌发酵滤液效果为最佳。     

The cumulative methane production from dairy cattle slurry can be explained by its volatile solid,temperature and length of storage

In order to refine the national estimate of methane emission from stored cattle slurry, it is important to comprehend the basic characteristics of methane production. Two dairy cattle slurries were obtained from livestock farms located in Hokkaido (a northern island) and Kyushu (a southern island). The slurries were diluted with water into three levels: undiluted, three times diluted, and 10 times diluted. Three hundred mL of the slurries were put into a bottle with a headspace volume of 2.0 L, which was filled with nitrogen gas and then sealed by butyl rubber. Four levels of temperature were used for incubation: 35, 25, 15 and 5 °C. The time course of the cumulative methane production per volatile solid (VS) was satisfactorily expressed by an asymptotic regression model. The effect of dilution on the methane production per VS was not distinctive, but that of temperature was of primary importance. In particular, higher temperature yields a higher potential production and a shorter time when the cumulative production reaches half of the potential production. The inclusive and simple models obtained in this study indicate that the cumulative methane production from stored cattle slurry can be explained by VS, temperature and length of storage.     

扑利旺防治柑橘红蜘蛛的应用技术研究

为探索扑利旺在柑橘红蜘蛛上的应用效果,开展了扑利旺防治柑橘红蜘蛛卵室内试验以及在不同虫口基数和不同温度条件下的田间试验。结果表明：扑利旺可明显抑制柑橘红蜘蛛卵的孵化,且孵化后幼螨存活率降低,并对幼螨、若螨和成螨均有较好的防治效果。生产使用中,在虫口基数低（2-3头/叶）或春季红蜘蛛发生期,推荐扑利旺200-300倍喷雾处理或扑利旺500倍+24%螺螨酯5000倍喷雾处理;而在虫口基数高（9-10头/叶）或秋季红蜘蛛繁殖高峰期,推荐扑利旺200-300倍+ 24%螺螨酯5000倍喷雾处理来控制柑橘红蜘蛛的危害。     

Effects of final dilution rate, sperm concentration and times for cooling and glycerol equilibration on post-thaw characteristics of canine spermatozoa

This study re-evaluated a protocol for cryopreservation of canine semen. Semen from 4 beagle dogs was pooled, concentrated by centrifugation and adjusted to increasing sperm concentrations by adding back seminal plasma. The prepared or original semen was diluted with an extender (Egg yolk-Tris-citrate-glucose) and cooled to 4 degrees C (cooling), followed by a second dilution with the same extender including glycerol, equilibrated at 4 degrees C (equilibration), then stored in liquid nitrogen. The semen was diluted for frozen samples having a fixed sperm concentration with increasing dilution rates or for those having the reverse combinations. Various dilution rates of 2.5-10 folds or sperm concentrations of 0.25-2.5 x 10(8)/ml had no significant effect on post-thaw sperm characteristics. When cooling was done for different times (0-26 hr) with glycerol equilibration for 1 hr, post-thaw characteristics were better at 2 and 3 hr of cooling, while various times for equilibration (0-4 hr) with cooling for 3 hr had no effect. These results suggest that different dilution rates and sperm concentrations within the ranges tested may not affect the post-thaw sperm characterisitics and that sufficient time for cooling may be essential but a specific equilibration time may not necessarily be required.     

Bactericidal effects of ozone at nonspermicidal concentrations.

A study was conducted to assess the use of ozone (O3) to control pathogens or contaminants of concern to animal breeders and regulatory officials. In separate experiments, samples of fresh bovine semen and Pseudomonas aeruginosa, Escherichia coli, or Campylobacter fetus subsp. venerealis were diluted with antibiotic-free milk (10(6) sperm and 10(6) organisms/mL of diluted semen), exposed in the previous day to a constantly monitored level of 5, 10, 15, or 20 micrograms/mL of O3 for 3-5 min. After 10 min at 30 degrees C, sperm motility was assessed and the samples cooled to 5 degrees C. Two and 18 h after the beginning of cooling, aliquots of each semen sample were evaluated for motility and cultured for organisms. Reductions were observed in P. aeruginosa and E. coli colony counts of 2 logs, and in C. fetus of 5 logs, after exposure for 2 h to O3 at a concentration of 5 micrograms/mL that had a moderate effect on sperm motility (reduction of 20%). Fewer than 100 colonies, i.e., a 4 logs reduction of all bacteria, were counted after dilution with ozonized-treated milk at 20 micrograms/mL of O3. However, this concentration of O3 reduced sperm motility by 50% 10 min after dilution. The results of these experiments indicate that a concentration and exposure time to O3 can be selected to reduce P. aeruginosa, E. coli, and C. fetus in contaminated bull semen diluted with milk while having only minimal effects on sperm motility.     

鸡传染性支气管炎病毒对新城疫病毒的干扰作用观察

为观察鸡传染性支气管炎病毒（IBV）HN99株对新城疫病毒（NDV）增殖的干扰作用,该试验采用不同浓度的IBV标准株M41和地方株HN99与鸡新城疫病毒（NDV）分别按不同接种顺序同胚增殖,利用病毒血凝试验（HA）测定NDV的效价,观察IBV对NDV的干扰作用,从而为检测IBV地方株HN99提供方法,也为同胚增殖两种病毒提供一系列的数据参考。试验结果表明,鸡传染性支气管炎病毒地方株HN99对NDV的干扰作用与其浓度和接种顺序有关。