口蹄疫病毒VP1基因的原核可溶性表达

针对重组原核表达载体表达目的蛋白以包涵体形式存在的问题,作者对影响外源蛋白表达的IPTG的浓度、温度、时间等因素均进行了探索,以观察VP1蛋白的可溶性情况。通过PCR技术将VP1基因克隆至pGEM-T载体中,然后将pET-28a(+)和pGEM-T载体分别进行双酶切,构建重组的pET-28a-VP1载体,将重组载体转化至BL21中,诱导后经SDS-PAGE检测可见约29 ku的目的蛋白条带。插入的外源目的蛋白主要以包涵体的形式存在,上清中的可溶性蛋白甚微。试验结果表明,这可能与蛋白本身的氨基酸组成有重要关联。     

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.     

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。     

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.     

罗非鱼湖病毒病传入风险分析

为评估罗非鱼湖病毒（tilapia lake virus,TiLV）跨境传入风险,按照OIE传入风险评估框架,从危害识别、风险评估及风险管理3个方面,开展了TiLV跨境传入风险分析,并提出了相应的风险控制措施。分析认为：活的敏感鱼类（包括亲鱼、鱼卵和鱼苗）,用做鲜活饵料的鱼类（野杂鱼）及冰冻和冰鲜的敏感鱼类（包括去除内脏的鱼肉）TiLV传入风险较高,而敏感鱼加工类产品、非敏感鱼类（包括活的和其他形式的产品）及运输活鱼的水、包装、运输工具和用具等传入风险较低。建议不从疫区进口高风险产品,对于低风险产品可以自由贸易,而对运输活鱼的水、包装、运输工具和用具等要进行强制常规消毒。     

鹰嘴紫云英头茬草生产性能的研究

通过对杨凌地区生长的鹰嘴紫云英（Astragalus cicer L.）的株高、株长、产量、草层结构、鲜干比、茎叶比等的研究表明：生长第二年第一茬初花期时刈割的鹰嘴紫云英平均株高达57 cm,株长达102 cm,鲜草产量达47 077.6 kg/hm^2,干草产量8 702.0 kg/hm^2,其茎、叶主要分布在0～40cm的草层中,茎、叶鲜重分别占到总重量的52.39%和31.88%,鲜干比为5.41：1,茎叶比为1.47：1,属典型的下繁型牧草,具产草量高、营养丰富、牧草品质好等优点,是值得推广的优良牧草.     

Dietary supplementation with Acanthopanax senticosus extract enhances gut health in weanling piglets

The present study was conducted to investigate the effects of Acanthopanax senticosus extract (ASE) as a dietary additive on gut health in weanling piglets by examining diarrhea frequency, intestinal microbiota and morphology. A total of 96 Duroc× (Landrace × Yorkshire) piglets weaned at 21 days of age with an average initial body weight (BW) of 5.6 ± 0.4 kg were randomly assigned to 3 treatment groups with 4 duplicates of 8 piglets each. The piglets were fed basal diet to which had been added 0 or 1 g/kg of ASE, or 0.7 g/kg antibiotics, respectively. Fecal consistence was monitored twice daily and the frequency of diarrhea was calculated. On day 21 after the initiation of supplementation, 8 piglets were randomly selected from each treatment group (2 piglets per pen) and slaughtered. The jejunum, ileum, colon and cecum were then excised and fixed in 10% neutral formalin solution to determine villus height and crypt depth, after their contents were collected to determine microbiota. The results showed that dietary supplementation with ASE increased (P < 0.05) the density of bacterial populations that co-migrated with Lactobacillus amylovorus, Lactobacillus salivarius, Bacillus subtilis, and Clostridium lituseburens, but decreased (P < 0.05) those co-migrating with Staphylococcus aureus, Salmonella typhimurium, Ruminococcus forques, and E. coli O157:H7 in the PCR-DGGE profiling analysis when compared with the control group. The villus height of the duodenum, jejunum and ileum increased (P < 0.05) by 14.8, 13.7 and 10.0%, while the crypt depth decreased (P < 0.05) by 17.9, 9.1 and 12.1%, respectively, in response to dietary ASE supplementation. Additionally dietary supplementation with ASE or an antibiotic decreased (P < 0.05) the frequency of diarrhea by 55.6 and 52.2%, respectively, compared with the control group. In conclusion, these findings suggest that dietary supplementation with ASE could regulate the microbiota composition and maintain a normal morphology of gut mucosa in weanling piglets, thereby decreasing diarrhea that resulted from weaning stress.