水稻胚乳双向电泳技术体系的优化分析

为比较同一杂交组合不同直链淀粉含量后代在灌浆期胚乳内蛋白表达差异,文章建立了一套适合灌浆期水稻胚乳蛋白的双向电泳技术,探讨了胚乳蛋白的提取、等电聚焦上样量的选择、胶条转移等双向电泳的一些关键技术。结果表明,一次溶解蛋白样品,TCA-丙酮法优于酚法蛋白提取,300μg上样量为最适上样量,能得到清晰、分离效果好、蛋白点数较多的图像。     

Effect of dietary supplementation of fish oil for lactating sows and weaned piglets on piglet Th polarization

The present study was designed to investigate the effect of dietary fish oil supplementation on piglet T helper cells (Th) polarization in relation to its impact on piglet serum interferon γ (IFN-γ) and interleukin 10 (IL-10) concentrations and splenic expression of Th1/Th2 characteristic genes. The diets of 18 gestating sows were supplemented with 7% lard (C) (n = 10) or 7% fish oil (T) (n = 8) from 10 d before parturition to weaning. At weaning, a split plot experiment was designed, 56 piglets, 28 each from sows fed with fish oil diet or lard diet, were divided into four groups of 7 replicates (one female and one castrated male per replicate) based on both sow diet during lactation and post-weaning piglet diet (C had 7% lard and T had 7% fish oil): CC, CT, TC, TT, and were fed the 7% fish oil or lard diet from day 35 to day 70. Serum concentrations of IFN-γ and IL-10, and Th1/Th2 related genes expression levels in spleen were measured and analyzed. The results showed that piglets fed with fish oil diet during post-weaning tended to have higher serum IFN-γ/IL-10 ratio (P = 0.09) than lard diet fed piglets. Lactation fish oil feeding increased splenic IL-12b, IL-12 receptor β2 (IL-12Rβ2), IL-2 and IFN-γ genes expression (P < 0.05 or P < 0.01) and post-weaning fish oil feeding increased splenic IL-12b (P = 0.06), IL-2 (P < 0.01) and IFN-γ (P = 0.08) mRNA expression than that in lard diet fed piglets at the end of this experiment. On the other hand, IL-4 gene expression (P = 0.01) in spleen was lower in weaned piglet from fish oil diet fed sows than that from lard diet fed sows. However, post-weaning piglets fed fish oil diet had higher splenic IL-4 (P = 0.06), IL-6 (P < 0.01) and IL-10 (P = 0.05) mRNA abundances than that fed with lard diet. These results indicated that dietary fish oil during lactation could increase Th1 polarization and accelerate immune maturation; while 7% fish oil in weaned piglets' diet was likely to increase Th2 cytokines expression.     

Effects of corn straw or mixed forage diet on rumen fermentation parameters of lactating cows using a wireless data logger

The objective of this study was to evaluate the effect of two different forage types on rumen fermentation parameters and profiles using a wireless data logger. Eight lactating cows were randomly assigned to one of two dietary treatments with a low forage diet with corn straw (CS) or a high forage diet with mixed forage (MF) as the forage source, respectively. Dietary physically effective neutral detergent fiber (peNDF) content was 11.3% greater in CS. Dry matter intake and milk fatty acid content decreased upon CS (P < 0.05). Ruminal pH, temperature and oxidation reduction potential (ORP) were monitored for 14 weeks. The CS group had significantly higher pH but lower temperature and ORP compared to MF (P < 0.01). With the CS diet regime, pH at the time before morning feeding, rumination and post‐ingestion were significantly higher than those in the MF group (P < 0.05). However, times with the ruminal pH below 6.0 and 5.8 were significantly reduced (P < 0.05), whereas ruminal pH below 5.6 tended to be lower (P = 0.07). The results indicated that rumen fermentation parameters were affected by forage types and dietary peNDF content might be predominant in ruminal pH regulation.     

不同过瘤胃淀粉和脂肪日粮对热应激奶牛生产性能和能量代谢的影响

试验研究了3种不同比例过瘤胃淀粉和脂肪日粮对泌乳中期热应激奶牛的影响。选择泌乳中期奶牛54头,根据产奶量、分娩时间和胎次随机区组分为低过瘤胃淀粉高脂肪组(PF101)、中过瘤胃淀粉中脂肪组(PF131)和高过瘤胃淀粉低脂肪组(PF161)3个处理组,处理组日粮的能量、蛋白相近。试验期间牛舍平均温湿指数为79.35。结果表明:PF161组较PF101组奶牛产奶量有所提高,乳蛋白含量较PF101组显著升高(P0.05),且泌乳净能有高于PF101组的趋势,PF161组奶牛乳中F∶P(脂肪蛋白比)显著低于PF101组(P0.05);PF161组奶牛血液中T4显著高于PF101组(P0.05),PF101组奶牛血液中胰高血糖素含量较PF161组和PF131组有升高趋势,总之高过瘤胃淀粉低脂肪日粮对泌乳中期热应激奶牛的泌乳性能和能量代谢均有一定改善作用。     

Model Optimization of Litter Size and Genetic Analyses of Model Parameters in Sows

According to two properties of the life cycle and to fluctuation with parities, four mathematical models, the Poisson cycle model, the cubic polynomial model, the modified quadratic polynomial model-I and the modified quadratic polynomial model-II, were used to fit the records of litter size in Jiangquhai sows. From the viewpoint of statistics and biological significance, the modified quadratic polynomial model-I was found to be the optimum model. A single trait animal model and DFREML procedures were further used to estimate the heritability values of optimum model parameters. The results show that the heritability values for the coefficients A and B and the heritability value for the acme of the model pure quadric curve are larger than the heritability value for the litter size. This suggests that selection for model parameters may be more effective than direct selection for litter size.     

禁食对生长期北京鸭腺胃ghrelin免疫阳性细胞的影响

本研究旨在观察禁食是否对生长期北京鸭腺胃ghrelin免疫阳性细胞生成有影响。用免疫组织化学的方法检测25、35、50日龄的北京鸭(已经禁食72 h)单位面积(mm2)腺胃组织中ghrelin阳性细胞数目。研究发现在这3个时期,不管是禁食组还是自由采食组,大量ghrelin阳性细胞多分布于腺胃深层复管状腺中;禁食组腺胃中gh-relin阳性细胞数目比自由采食组极显著增加(P0.01)。结果提示:禁食是影响腺胃产生有活性ghrelin的重要因素。     

猪圆环病毒2型云南株分离鉴定及ORF2序列分析

本研究根据GenBank中已经发表的猪圆环病毒2型基因序列,设计了2对PCV2特异性引物,从疑似断奶仔猪多系统衰竭综合征(postweaning multisystemic wasting syndrome,PMWS)感染病例中检测出3株云南省PCV2流行株,通过ve-ro细胞分离病毒,并用透射电子显微镜观察到了约17nm的病毒样粒子的存在。经同源性比较分析,3个分离株之间核苷酸同源性为99.5%～99.8%,与甘肃省PCV2分离株核苷酸同源性最低(90.7%),浙江省分离株核苷酸同源性最高(99.7%),与南美洲分离株核苷酸同源性最低(92.0%),瑞典分离株核苷酸同源性最高(99.4%),这可能与云南省猪种引进有关。     

稳定表达猪圆环病毒2型Cap蛋白的CHO-K1细胞系的建立及免疫原性分析

建立高表达PCV2-Cap蛋白的CHO-K1悬浮稳转细胞系,鉴定蛋白的免疫原性,为开发新型且有效防治猪圆环病毒的亚单位疫苗奠定基础。构建重组质粒pEE12.4-PCV2-Cap,转染CHO-K1细胞,通过加压筛选,有限稀释,细胞悬浮驯化及Western blot检测得到高表达Cap蛋白的悬浮稳转单克隆细胞株,并对该细胞株进行发酵,纯化得到的目的蛋白,通过小鼠免疫验证其免疫原性。结果表明PCV2-Cap蛋白能够在CHO-K1细胞中正确表达;发酵过程中活细胞密度高达6×10~6个·mL^-1,细胞活力在80%以上,PCV2-Cap蛋白表达量约为370 mg·L^-1;利用间接ELISA检测小鼠免疫后血清中的抗体水平,证明了利用CHO-K1细胞生产的Cap蛋白具备良好的免疫原性。本研究成功构建了表达PCV2-Cap蛋白的CHO-K1悬浮稳转细胞系,并进一步对目的蛋白进行免疫原性分析,为猪圆环病毒亚单位疫苗的开发打下扎实的基础。     

Effect of system of feeding and watering on performance of lactating sows

An experiment was conducted to determine the effects of ad libitum access to feed and water and the option to mix feed and water, all in the same feeder, on the performance of multiparous lactating sows. Feed and water were made available to sows using a self-fed wet/dry (SFWD) or a hand-fed (HF) feed-water system. In the SFWD system, feed and water were dropped into a common trough area of the feeder. The sow determined when and how much of each was dropped. With feed falling onto the flat area of the bottom of the SFWD feeder trough and water falling into the shallow bowl area, and with the 2 areas seamlessly connected, the sow also determined the wetness of the feed consumed. In the HF system, sows were given dry feed twice daily in a J-shaped feeder that was independent of the sow's water source. Sows (n = 114) were assigned to treatments based on parity and genotype. Total feed disappearance per sow during lactation (20 +/- 0.2 d) was greater (P < 0.01) with the SFWD system than with the HF system (120 vs. 110 +/- 4.1 kg, respectively). The SFWD sows had greater (P < 0.01) BW gains during lactation than HF sows (6.2 vs. 0.6 +/- 1.85 kg, respectively). Backfat depth change during lactation did not differ (P = 0.37) between treatments. Likewise, percentage of sows displaying estrus by d 11 post-weaning did not differ (P = 0.51). Piglet weaning BW was greater (P < 0.01) with the SFWD system than with the HF system (6.63 vs. 6.12 +/- 0.22 kg, respectively). Sow average daily water intake and total feed wastage during lactation did not differ (P > 0.66) between treatments. However, sows with the SFWD system wasted less water (P < 0.01) than those with the HF system (15 vs. 232 +/- 12 L, respectively). From a commercial swine production perspective, the difference in waste water volume would result in a significant variation in costs associated with manure storage and distribution. In conclusion, use of a SFWD feed-water system in lactation, which provides sows choices of when to eat, how much to eat, and if dry feed should be mixed with water during consumption, enhances sow appetite, improves litter growth performance, and wastes less water than a HF feed-water system.     

仔猪水肿病蜂胶佐剂多价灭活苗的研制Ⅱ免疫保护期和疫苗保存期试验

对仔猪水肿病蜂胶佐剂多价灭活苗进行了免疫产生期、免疫持续期和疫苗保存期试验。研究表明 :仔猪接种该疫苗 5～ 7d后便产生免疫力 ,到第 9天时抗体产生水平已经很高 ;以免疫仔猪攻毒后的存活率不低于 95%为标准 ,疫苗的免疫持续期可达 9个月以上 ;以物理性状合格和免疫仔猪攻毒后的存活率不低于95%为标准 ,疫苗置 4℃时保存期为 1 8个月 ,2 5℃保存 9个月为宜