Generation of a genotype VII Newcastle disease virus vaccine candidate with high yield in embryonated chicken eggs

To generate a genotype VII Newcastle disease virus (NDV) vaccine with high yield in embryonated chicken eggs, we selected genotype VII NDV strain JS5/05, which possesses a high virus titer in embryos as the parental virus. Using reverse genetics, we generated a genetically tagged derivative (NDV/AI4) of JS5/05 by changing the amino acid sequence of the cleavage site of the F0 protein. Pathogenicity tests showed that NDV/AI4 was completely avirulent. NDV/AI4 was genetically stable and replicated efficiently during 10 consecutive passages in embryos. More importantly, serologic assays showed that oil-emulsion NDV/AI4 induced higher hemagglutination inhibition (HI) titers against the prevalent virus than oil-emulsion LaSota vaccine in chickens and geese. Moreover, NDV/AI4-induced HI titers rose faster than those elicited by LaSota in chickens. Both NDV/AI4 and LaSota provided protection against clinical disease and mortality after the challenge with the genotype VII NDV strain JS3/05. However, NDV/AI4 significantly reduced virus shedding from the vaccinated birds compared to LaSota. Taken together, these results suggest that NDV/AI4 can provide better protection than LaSota and is a promising vaccine candidate against genotype VII NDV.     

冷冻干燥法制备速溶植酸酶的研究

采用毕赤酵母发酵制备酶液,膜超滤浓缩、硫酸铵盐析法沉淀酶蛋白,结合冷冻干燥法制得速溶型植酸酶。结果表明,对酶活力达19 500U/ml的发酵液进行浓缩后,以70%饱和度的硫酸铵进行盐析,分离后的酶蛋白经过冷冻干燥得到活力为500 000U/g的植酸酶,速溶性好,溶解后清亮透明。酶的冻干粉50℃保存90d内酶活力仍能维持96%以上的活力。为高酶活速溶植酸酶的工业化生产提供了一条可行的思路。     

Effect of mSOF and G1.1/G2.2 Media on the Developmental Competence of SCNT‐Derived Bovine Embryos

The objective of this study was to compare the effect of two culture media: modified synthetic oviductal fluid (mSOF) and G1.2/G2.2, on the developmental competence of bovine somatic cell–cloned embryos. Cloned embryos were produced by transferring adult skin fibroblasts into enucleated MII oocytes. After activation, the reconstructed embryos were randomly allotted to either mSOF or G1.2/G2.2 for culture (the embryos were transferred from G1.2 to G2.2 on days 3 of culture). The development competence of cloned embryos in these two culture systems was compared in terms of cleavage rate, blastocyst formation rate and apoptosis cell number in day 7 blastocyts. To investigate the in vivo developmental competence of cloned embryos in the two culture systems, a total of 87 and 104 blastocysts derived from mSOF and G1.2/G2.2 medium groups were transferred individually to recipient Angus cows, respectively. No differences were observed in terms of cleavage rate, day 7 blastocyst rate and blastocyst cell number between these two culture systems. However, the day 6 blastocyst formation rate was significantly higher in G1.2/G2.2 than that in mSOF. In addition, blastocysts cultured in mSOF have a higher percentage of apoptotic blastomeres compared to those in G1.2/G2.2 (8.5 ± 1.2 vs 16.8 ± 1.5, p < 0.05). Although difference in pregnancy rate was not observed 40 days after embryo transfer, significantly higher pregnancy rate was observed in G1.2/G2.2 group after 90 days of embryo transfer (12.4% vs 37.5%, p < 0.05). Moreover, calving rate was significantly improved in G1.2/G2.2 group compared to mSOF group (27.9% vs 6.7%, p < 0.05). In conclusion, our results indicate that G1.2/G2.2 can improve developmental competence of bovine SCNT embryos both in vitro and in vivo, which is more suitable for culture of bovine SCNT embryos than mSOF medium.     

中草药方剂对肉仔鸡脂类代谢的影响

选取225只1日龄艾维茵肉鸡,随机分为5组,每组3个重复,每个重复15只鸡,研究中草药方剂对肉仔鸡脂类代谢的影响。基础日粮为玉米-豆粕型,对照组饲喂基础日粮,试验组分别在基础日粮中添加0.5%、1.0%、1.5%、2.0%的中草药复方添加剂,进行为期6周的饲养试验。结果表明:日粮中添加该方剂显著影响了肉仔鸡血液中总胆固醇(TC)、甘油三酯(TG3)、高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇(LDL-C)的含量,且随着中草药添加水平的升高,TC、TG3、LDL-C含量有下降的趋势,而HDL-C则显著升高,并且能显著降低腹脂率、肝脂含量和肌肉中胆固醇含量。     

抗球虫中药散剂常青散的亚急性毒性试验

为了测定抗球虫中药散剂常青散的亚急性毒性反应,取昆明小鼠为试验动物,随机分组,以临床应用剂量的2、4、8倍饲喂给药,确定常青散在28天内对小鼠的亚急性毒性反应。结果表明各试验组小鼠全部健活,体征、体重、外观、行为、采食量及饲料利用率、脏器系数、血液生理生化指标、脏器结构均无异常。常青散对受试动物未见有明显的亚急性毒性反应。     

银狐垂体细胞系构建研究

分别采用组织块法和消化法2种接种方法进行银狐垂体细胞系构建研究。在消化法中使用不同浓度的胰蛋白酶来检验效果,同时在原代培养期使用反复差速贴壁法纯化,传代培养期结合使用反复差速贴壁以及两步消化法进行处理。结果表明：组织块法接种后成纤维细胞生长明显,原代培养后期成纤维细胞已为主要细胞;降低胰蛋白酶浓度能降低对细胞损伤,但消化时间会增长;采用消化法接种在抑制成纤维细胞生长方面效果好于组织块法。纯化效果上,结合反复差速贴壁法与两步消化法处理后获得了较好的效果,在原代培养时期能较好去除成纤维细胞并通过持续使用该方法可在传二代后获得较高纯度的细胞,纯度能达到80%以上,能够建立细胞系。     

奶牛早期妊娠诊断研究进展

<正>奶牛配种(或受精)后不久,必须进行早期妊娠诊断,以便加强怀孕母牛饲养管理,保证顺利分娩或对未孕母牛及时配种,缩短产仔间隔,提高繁殖效率。生产实践中广泛应用的直肠检查等方法,尽管简单易行,不需特     

生鲜乳中添加皮革水解蛋白的测定方法研究

L-羟脯氨酸是皮革水解蛋白构成的特征氨基酸,因此鉴定L-羟脯氨酸就是鉴定皮革水解蛋白。建立UV法测定生鲜乳中皮革水解蛋白,L-羟脯氨酸浓度在0μg/mL～2.0μg/mL范围呈良好的线性关系,R=0.9998。L-羟脯氨酸平均回收率为100.2%,猪皮粉平均回收率为98.1%,皮革粉平均回收率为99.9%。该方法回收率高,检出限低,工作曲线稳定,重复性好。     

鲈鲤肌肉脂肪酸组成分析

对鲈鲤肌肉脂肪酸进行测定,结果显示:鲈鲤肌肉中含有7种脂肪酸,饱和脂肪酸有3种,不饱和脂肪酸有4种。3种饱和脂肪酸(SFA)总量为28.22%,4种不饱和脂肪酸(UFA)为71.75%,以不饱和脂肪酸中油酸C18∶1含量最高,为44.28%。鲈鲤鱼体肌肉脂肪酸主要由油酸C18∶1、棕榈酸C16∶0、亚油酸C18∶2和棕榈油酸C16∶14种脂肪酸组成,这4种脂肪酸质量分数总和占脂肪酸总质量分数的93.58%。鲈鲤肌肉脂肪酸种类组成与其它几种淡水鱼类比较,存在明显差异。     

Effect of selenium-enriched malt on hypoglycemia and regulatory hormones in diethylnitrosamine-induced hepatocarcinoma SD rats

One hundred and ninety-three Sprague-Dawley (SD) rats (average body weight being 100–120 g) were randomly divided into 5 groups (I–V). Animals in group I and group II served as the negative control and positive control, respectively, and both received 0.1 mg/kg selenium (Se) from sodium selenite. Animals in groups III–V were fed with Se from Se-enriched malt (SEM) supplemented diets (0.3, 1 and 3 mg/kg, respectively). Simultaneously, hepatocarcinoma were induced in groups II–V by diethylnitrosamine (DEN) solution (100 mg/L) at the dosage of 10 mg/kg body weight every day as drinking water for 16 weeks, then sterilized water for a further two weeks. Rats of group I drank sterilized water during the whole experimental time. At 4th, 8th, 12th, 16th week, five rats in each group were then sacrificed by cervical decapitation. At the termination of the study, at 18th week, the surplus rats were sacrificed by cervical decapitation. Feed was withheld from the rats for 12 h before sampling. The values of plasma glucose at different sampling times were measured. The values of the hormones in plasma related to plasma glucose metabolism, including insulin, glucagon, insulin-like growth factors-II (IGF-II), and the ratios of insulin/glucose (IGR₁), insulin/glucagon (IGR₂) and glucagon/glucose (GGR) were determined. At the same time, the correlation of plasma glucose concentrations related to hormones was statistically analyzed. The results indicated that the values of plasma glucose, insulin, glucagon and GGR in the groups treated with DEN were decreased significantly as compared with that of the negative control group, however, the values of IGF-II and IGR₂ were increased significantly. SEM showed a significant effect in suppressing the decreased of plasma glucose and glucagons, and delaying the increased of IGF-II and IGR₂ in the DEN-induced hepatocarcinoma rats. The plasma glucose concentrations revealed a significant relation to the hormones. In conclusion, SEM could reduce the development of hypoglycemia in the DEN-induced hepatocarcinoma rats by regulating the relative levels and balances or proportions of hormones.