动物疫病检测实验室规范化管理初探

随着我国社会主义市场经济的不断完善和外向型畜牧业的迅速发展．动物疫病检测实验室的规范化管理越来越显得尤为重要．建立健全实验室质量体系．依法进行计量认证和实验室认可．就成为动物疫病检测实验室规范化管理的必要途径。青岛市畜牧兽医工作站依法承担着全市动物疫病检测工作．为了有效实施规范化管理．从2004年4月份开始建立质量体系．并于2004年11月同时通过了中国实验室国家评审委员会组织的实验室认可和山东省质量技术监督局组织的计量认证现场评审．分别于2004年12月和2005年1月取得了认可、认证证书。     

重组逆转录病毒介导的肿瘤坏死因子基因转移与哺乳动物细胞表达

以构建的重组肿瘤坏死因子逆转录病毒载体ｐＺＩＰＴＮＦ ＤＮＡ转染Ｃｏｓ７细胞，获得了ＴＮＦα暂时性表达。采用ＤＮＡ－磷酸钙共沉淀法，将ｐＺＩＰＴＮＦ ＤＮＡ导入了单向性逆转录病毒包装细胞ψ２，在Ｇ４１８选择下，克隆出了ψ／ｐＺＩＰＴＮＦ细胞系。该细胞可产生重组ＴＮＦ逆转录病毒。本研究利用ψ／ｐＺＩＰＴＮＦ细胞分泌的重组逆转录病毒连续传代感染ψ２或ＰＡ３１７细胞以及病毒交替感染ψ２和ＰＡ３１７细胞，     

Shifts in disease resistance induced by growth regulators

Experiments on the effect of various compounds with plant growth regulating activity led to the hypothesis that conditions inhibitory to indoleacetic acid (IAA) action or leading to a decrease in the IAA level in cucumber seedlings would be unfavourable for the development of cucumber scab, caused byCladosporium cucumerinum. Susceptibility decreased as the result of treatment with growth retardants, which cause an increase in IAA-oxidase activity in the plants, whereas application of indoleacetic acid or compounds expected to decrease the rate of oxidative breakdown of IAA in the seedlings resulted in an increase of susceptibility. Growing the plants under various periods of illumination also influenced both the susceptibility of the plants and the IAA-oxidase activity in the hypocotyl tissue.To obtain further information about the relation between the increase in resistance and the increase in the rate of IAA oxidation, the effect of one compound, viz. L-threo-β-phenylserine (abbr. phenylserine) was studied in more detail. The rate of oxidative breakdown of IAA was increased in extracts of hypocotyls and cotyledons of plants treated with phenylserine as compared with extracts of tissue of untreated plants. Extracts of phenylserine-treated plants were found to contain a higher cofactor and/or a lower inhibitor content than those of control plants. The substances involved were not identified, but this result may indicate a shift in the concentrations of various phenols in the tissue.The question must remain open whether a shift in the concentration of phenols some-how effected by phenylserine treatment determines both degree of susceptibility and rate of IAA oxidation independently, or whether the rate of IAA breakdown and the resulting change in IAA content in the plants is directly related to the degree of susceptibility.     

New potyvirus isolated from <Emphasis Type="Italic">Cryptotaenia japonica</Emphasis>

 A potyvirus, for which the name Japanese hornwort mosaic virus (JHMV) is proposed, was isolated from Japanese hornwort plants (Cryptotaenia japonica) with mosaic disease symptoms. The virus was used to inoculate mechanically 34 plants belonging to 33 species of 10 families. Of these species seven from two families were infected. Faint chlorotic spots appeared on the inoculated leaves of Chenopodium quinoa and C. amaranticolor, but no systemic infection occurred in these plants. JHMV systemically infected only Umbelliferae plants; they did not infect 26 other species in eight families. JHMV was transmitted in a nonpersistent manner by aphids (Myzus persicae). The virus was a flexuous rod-shaped particle about 750 nm in length. Sequencing the nucleotides in the 3′ terminal region of JHMV revealed that the coat protein contains 280 amino acids with a molecular mass of 32.2 kDa. The nucleotide sequence of the coat protein of JHMV had the highest similarity with that of Zantedeschia mosaic virus (83.3%) compared to those of other potyviruses (57.0%–64.9%). An antiserum against JHMV reacted strongly with JHMV and weakly with Potato virus Y. These results indicate that JHMV is a new potyvirus. Received: September 9, 2002 / Accepted: November 7, 2002 RID="*" ID="*" The nucleotide sequence determined in this work appears in the DDBJ/EMBL/GenBank nucleotide sequence databases with the accession number AB081518     

Efficacy of live virus vaccines against infectious laryngotracheitis assessed by polymerase chain reaction-restriction fragment length polymorphism

The efficacy of four different commercial live vaccines (vaccines A, B, C, and D) against the infectious laryngotracheitis virus (ILTV) was assessed in specific-pathogen-free (SPF) chickens. SPF chickens were vaccinated intraocularly at 6 wk old with ILTV live vaccines and were challenged intratracheally with the N91B01 strain of virulent Korean ILTV 2 wk after vaccination. The immunity against ILTV live vaccines was assessed by the incidence of latent infection by the challenge virus in the chickens' tracheas and trigeminal ganglia, the reisolation rate of the challenge virus, and the clinical signs in the chickens challenged with the N91B01 strain of ILTV. The latent infection in chickens was assessed by nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Our data showed that the clinical signs and challenge virus isolation were negative in all chickens receiving four difference commercial ILTV live vaccines. The viral DNA of the vaccine strain, but not that of the challenge virus, was detected in chickens vaccinated with vaccine A by nested PCR-RFLP. The viral DNAs of both the vaccine and challenge strains were detected from chickens vaccinated with vaccines B, C, and D. This study showed that only vaccine A can protect chickens from latent infection with the field virulent ILTV. We speculate that the efficacy of infectious laryngotracheitis live vaccines to protect chickens from latent infection with virulent ILTVs can be assessed by nested PCR-RFLP analysis.     

Effects of nutrient supply and dietary bulk on O2 uptake and nutrient net fluxes across rumen,mesenteric- and portal-drained viscera in ewes

We assessed the effects of nutrient supply and dietary bulk, both increasing with hay intake, on O2 uptake and nutrient net fluxes across the portal-(PDV) and mesenteric- (MDV) drained viscera, and the rumen in adult ewes. Four ewes, fitted with a ruminal cannula, with catheters in the mesenteric artery, the portal, mesenteric and right ruminal veins, and with a blood flow probe around the right ruminal artery, were used in a 4 x 4 Latin square design. Treatments consisted of 500 g DM/d hay (LL, low bulk and low nutrient supply), 500 g DM/d hay + infused nutrients (LH, low bulk and high nutrient supply), 750 g DM/d hay + infused nutrients (MH, medium bulk and high nutrient supply), and 1,000 g DM/d hay (HH, high bulk and high nutrient supply). Infused nutrients consisted of volatile fatty acids (VFA) and casein dissolved in salts and infused continuously in the rumen to provide the same amount of metabolizable energy (7.6 MJ/d) and digestible protein (63 g/d) for LH, MH, and HH. Both increases in bulk and nutrient supply increased O2 uptake in the MDV and PDV. Dietary bulk stimulated mainly blood flow, whereas nutrient supply stimulated mainly O2 extraction rate. The O2 uptake by the rumen was not significantly affected by hay intake, although blood flow increased due to nutrient supply. Increase in hay intake had no effects on portal net release of lactate and net uptake of glucose but increased VFA, 3-D-hydroxybutyrate, ammonia, and amino acids (AA) net release and urea net uptake across PDV. The increase in portal nutrient net fluxes with hay intake was entirely related to the increase of nutrient supply for VFA, 3-D-hydroxybutyrate, ammonia, and urea, irrespective of the amount of casein infused for AA. Dietary bulk had no effect on total energy net release in the portal vein. We conclude that despite the increase in portal O2 uptake, increasing dietary bulk had no significant impact on portal recovery of energy. In ruminal tissues, which were the main site of energy absorption, O2 uptake appeared low and was not sensitive to dietary manipulation. In contrast, in mesenteric tissues, which contribute poorly to energy absorption with forage diets, O2 uptake appeared high and very sensitive to dietary manipulation.     

不同性别生长猪所需最佳苏氨酸与赖氨酸比率

本研究通过调查苏氨酸与赖氨酸的比率对猪生长性能、表观可消化能和血液尿素氮 (BUN)浓度的影响 , 估测生长阉公猪和小母猪所需最佳苏氨酸与赖氨酸的比率 . 试验采用 150头杂交猪 (长白×约克夏×杜洛克 , 平均体重为 16.75± 0.45 , 其中阉公猪 75头 , 小母猪 75头 ), 按 2× 3析因设计 , 随机分成 6个处理 阉公猪日粮含赖氨酸 1.12% , 按苏氨酸与赖氨 酸比率为 50% 、 60% 和 70% 分成 3个处理 小母猪日粮含赖氨酸 1.33% , 按苏氨酸与赖氨酸比 率为 50% 、 60% 和 70% 分成 3个处理 . 经整个试验期 (体重从 16至 56 ). 在平均日增重、平均 日采食量和饲料转化率方面 , 动物性别和苏氨酸与赖氨酸比率之间无相互影响 . 在生长性能 方面 , 有明显的性别影响 与小母猪相比 , 阉公猪表现出较好的生长性能 , 消耗较多的饲料 (P< 0.01)以及生长的较快 (P< 0.01). 随着苏氨酸与赖氨酸比率的增大 , 阉公猪的 ADG和 FCR有增加的倾向 , 但差异不显著 . 在除粗灰分 (CA)外的可消化性养分方面 , 动物性别和苏氨 酸与赖氨酸比率之间无相互影响 . 在性别之间 , 除小母猪对钙具有较高的可消化性 (P< 0.01)外 , 可消化性养分没有存在的差异 . 而日粮苏氨酸与赖氨酸比率对可消化性养分无影响 . 可消化性必需氨基酸 (EEA)、非必需氨基酸 (NEAA)和总氨基酸 (TTA)的平均值没有受到性别 和苏氨酸与赖氨酸比率的影响 . 没有发现性别和日粮中苏氨酸与赖氨酸比率相互影响的证据 . 在性别之间 , 小母猪的总 BUN浓度比阉公猪低 (P< 0.05). 可以得出结论 " 阉公猪日粮 (赖氨 酸水平为 1.12% )中 , 苏氨酸与赖氨酸比率为 70% , 小母猪日粮 (赖氨酸水平为 1.33% )中 , 苏 氨酸与赖氨酸比率为 60% , 较其它比率的苏氨酸与赖氨酸 , 分别获得较好的生长性能、养分 利用和较低的 BUN浓度 .     

乳中黄嘌呤氧化还原酶的研究进展

黄嘌呤氧化还原酶属于含钼脱氢酶黄素蛋白家族，是组成乳脂肪球膜的主要蛋白，通过硫酸胺分级沉淀和DEAE-Sepharose柱可从乳脂肪球膜中纯化。XOR作为乳脂肪球膜的膜结合蛋白，在脂肪滴的包裹和分泌中起到重要的结构作用，并且在肠道能够起到一定的抗菌作用。     

猪瘟免疫接种抗体效价检测试验

1前言猪瘟只有一个血清型,免疫注射是防止猪瘟的重要手段。我国猪瘟兔化弱毒苗自1956年研制成功以来,在全国各地广泛应用,证明安全有效。免疫疫苗24小时后,即可抵抗自然感染,第3天产生可靠的免疫力;在疫区对体温及体况正常的猪用此苗紧急接种,能迅速终止疫情发展。断奶仔猪接种疫苗后产生的免疫期至少1年。幼年仔猪的免疫期视其由初乳获得的母源抗体而异,有的达1年,有的不及1个月。仔猪获得的母源抗体,一般可维持30～45d,对主动免疫力的产生有干扰作用。如免疫过早,因母源抗体水平高,产生的免疫力差,免疫期很短,因而有接种后不久发生猪瘟的…