饲粮添加氨基酸锌对育肥羊屠宰性能、肉品质及血液和组织中微量元素含量的影响

本试验旨在研究氨基酸锌对育肥羊屠宰性能、肉品质及血液和组织中微量元素含量的影响。选择4月龄、体重为(29.00±0.61)kg的健康滩湖杂F1代公羊48只,随机分为4组,每组12只。对照组饲粮锌含量为52.80 mg/kg(硫酸锌),其中不含硫酸锌的锌含量为18.02 mg/kg,试验Ⅰ组、试验Ⅱ组、试验Ⅲ组试验饲粮锌含量分别为对照组饲粮的1/2、1和2倍(氨基酸锌),预试期7 d,正试期60 d。结果表明:饲粮添加氨基酸锌显著提高了育肥羊平均日增重(P<0.05),以试验Ⅱ组平均日增重最高,达到283.59 g/d。饲粮添加氨基酸锌对育肥羊屠宰性能、肉质理化性状无显著影响(P>0.05),但随着饲粮氨基酸锌添加量增加,试验组肉骨比、背膘厚、眼肌面积先升高后降低,GR值、肌内脂肪含量升高,剪切力降低;试验组血液锌含量均高于对照组,在同等锌添加量条件下,试验Ⅱ组血清锌含量极显著高于对照组(P<0.01);各组间背最长肌、肾脏、心脏、肝脏及肺脏组织的锌含量差异不显著(P>0.05),但氨基酸锌添加量过高会影响肌肉、肾脏、心脏、肺脏中铁沉积及肝脏中铜沉积。综上所述,饲粮添加氨基酸锌可促进育肥羊生长,改善肉品质;在锌含量为18.02 mg/kg的基础饲粮中添加52.80 mg/kg锌(氨基酸锌)效果最佳。     

基于随机森林的高寒草地地上生物量高光谱估算

草地地上生物量（Aboveground biomass,AGB）是衡量草地生产力的关键因素,准确测定草地AGB具有重要意义。高光谱因具有时效性强、不破坏草地等特点被广泛用于草地生理生态指标的测定。本研究提取和计算了海北试验站高寒草地冠层的原始光谱（Original spectrum,OR）反射率、一阶微分光谱（First derivative spectrum,FD）反射率、光谱位置面积参数（Spectral parameters of spectral position and area,PA）和植被指数（Vegetation indices,VI）4种不同类型的特征变量,使用连续投影算法（Successive projections algorithm,SPA）和递归特征消除算法（Recursive feature elimination,RFE）进行特征选择,采用随机森林算法（Random forest,RF）构建草地AGB估测模型。结果表明：在由4种特征变量分别构建的草地AGB估测模型中,基于VI的RF模型精度最高（测试集R²=0.70,RMSE=557.87 kg·ha^-1）,实测AGB与估测AGB的线性R²达到0.72;不同类型特征变量组合构建的草地AGB估测模型中,PA+VI组合的RF模型精度最高（R²=0.71,RMSE=548.97 kg·ha^-1）,实测AGB和估测AGB的线性R²达到0.73。     

氯前列醇钠、产前背膘厚及便秘对后备母猪同期分娩效果的影响

试验旨在研究氯前列醇钠、产前背膘厚、便秘对后备母猪同期分娩集中度和生产性能的影响,并进一步探讨母猪同期分娩的最优方案。选取后备母猪共295头,根据妊娠期110 d时P2点处的背膘厚,随机分为3组：对照组（CON组：0.9%生理盐水）、低剂量氯前列醇钠组（LPG组：0.1 mg氯前列醇钠）、高剂量氯前列醇钠组（HPG组：0.2 mg氯前列醇钠）,于妊娠期114 d进行阴户部注射诱导分娩。结果表明,与对照组相比,高、低剂量氯前列醇钠均可显著缩短母猪从注射到分娩的时间间隔（P<0.05）,且两试验组间差异不显著（P>0.05）。但是,LPG组后备母猪在注射氯前列醇钠后48 h内的分娩率高达95%,并且白天分娩率达75.7%。此外,LPG组后备母猪的产仔间隔也显著短于CON组（P<0.05）。同时本研究发现,背膘厚在16~20 mm范围内的后备母猪从注射到分娩的时间间隔显著短于背膘厚≤15 mm的后备母猪（P<0.05）;便秘指数0~1.0组后备母猪的产程和产仔间隔均显著大于便秘指数1.1~2.0组及2.1~3.0组的后备母猪（P<0.05）。进一步回归分析发现,母猪背膘厚分别与产程、总产仔数、活仔数存在极显著线性相关（P<0.01）,母猪便秘指数分别与产程、产仔间隔、总产仔数、活仔数存在极显著线性相关（P<0.01）。综上所述,在妊娠期114 d对后备母猪阴户部注射0.1 mg氯前列醇钠诱导分娩,不仅可显著加快母猪分娩的进程,而且还可大大节约药物成本。此外,母猪产前背膘厚和便秘均会影响母猪同期分娩的效果,应使母猪分娩前保持最佳的背膘厚（16~20 mm）,且避免母猪便秘,方可保证同期分娩的顺利实施。     

气肿疽梭菌CctA基因的原核表达及其间接ELISA检测方法的建立

研究旨在原核表达气肿疽梭菌细胞毒素A （CctA）基因,用纯化的重组蛋白建立其间接ELISA检测方法,a。利用大肠杆菌密码子的偏爱性优化CctA基因序列,克隆至原核表达载体pET-28a （+）,双酶切鉴定原核表达质粒pET28a-CctA并测序。将重组质粒转入大肠杆菌,经IPTG诱导得到高表达的重组CctA包涵体蛋白。包涵体蛋白变性后经镍（Ni）柱纯化,SDS-PAGE检测其纯化效果。以豚鼠抗气肿疽梭菌抗血清为一抗,用Western blotting方法检测重组CctA蛋白的反应原性。用棋盘滴定法建立间接ELISA检测方法,以复性后的重组CctA蛋白作为检测抗原,摸索抗原包被浓度、封闭液的种类及浓度、抗体的最适稀释度和反应条件等。选用3批气肿疽灭活疫苗免疫豚鼠后采集血清,同时用攻毒和间接ELISA两种方法验证疫苗的免疫效力。质粒双酶切结果显示,得到大小约853 bp的条带,与预期相符,且测序结果正确。SDS-PAGE结果表明,成功表达并纯化大小为35 ku的重组CctA蛋白。Western blotting结果显示,豚鼠抗气肿疽梭菌抗血清与重组CctA蛋白具有良好的反应原性。建立的间接ELISA法的最适条件为：抗原的包被浓度为0.5 μg/mL,于4 ℃包被过夜;封闭液选择10%胎牛血清,37 ℃孵育2 h;二抗的稀释度为1：8 000;室温避光显色10 min后终止反应,测定D_{450 nm}值。当P/N>4.6时,间接ELISA法检测结果与豚鼠攻毒试验结果拟合度较好。本研究成功表达CctA基因并纯化了重组CctA蛋白,建立的以重组CctA蛋白为检测抗原的间接ELISA检测方法,有望成为气肿疽灭活疫苗免疫效果验证的替代方法。     

拷贝数变异在畜禽中的研究进展

拷贝数变异（copy number variation,CNV）具有多种形式的变异结构,在品种多样性、生物进化和疾病相关性等研究中起着重要作用,并具有片段长度大、覆盖范围广等特点。随着分子生物学的发展及DNA测序技术的日渐成熟,人们对遗传变异的研究不断向DNA分子水平深入,多态性标记在畜禽育种中已逐渐成为动物育种研究的趋势和主流。由于CNV对基因的调控和表达所造成的影响更为显著,因此,CNV在重要畜禽中的研究越来越多。目前,已检测出大量有关畜禽重要经济性状的基因序列变异,并有许多研究均表明CNV与动物的重要经济特征及疾病的发生有关。笔者主要通过参考国内外相关的研究报道,简述了CNV的相关研究背景、概念、突变机制,归纳总结了CNV对牛、羊、猪、鸡的经济性状、繁殖性状和疾病调控的影响,以期通过对这些重要畜禽的基因组学研究揭示其适应性遗传机理和表型性状差异的遗传基础,开发相应的分子遗传标记,为畜禽的标记辅助选育提供理论基础。     

Effect of Se and VE Supplement on Semen Quality,Antioxidant Enzyme Activities and Heat Shock Protein Expression of Goat in High Temperature Season

This experiment was conducted to study the effect of selenium and vitamin E supplement on semen quality, antioxidant enzyme activities and heat shock protein expression of goat in Hainan high temperature season.16 adult Hainan Black goat with good health and approximate weight were randomly divided into 4 groups, fed with basal diet(control group), basal diet+0.5 mg/kg Se(Se group), basal diet+100 mg/kg VE(VE group), and basal diet+0.5 mg/kg Se+100 mg/kg VE(Se+VE group), respectively.The experimental period was 93 d.Semen samples were collected in the last week of the experiment on two consecutive days.The semen quality, antioxidant enzyme activities and heat shock protein expression were analyzed.The results showed that compared with control group, the ejaculate volume was not significantly affected by Se or VE supplement(P>0.05).Sperm density and sperm motility were increased significantly by Se and VE supplement(P<0.05), and the abnormal rate was decreased extremely significantly(P<0.01).The goats fed with Se and VE also had higher activities of GSH-Px(P<0.01), SOD(P<0.05), CAT(P<0.05) and T-AOC(P<0.01), and lower MDA concentration in seminal plasma(P<0.05).The relative expression levels of HSP70 and HSP90 mRNA in supplement groups were decreased extremely significantly(P<0.01).However, there were some certain differences between the Se and VE supplement groups on semen quality and heat shock protein expression.In conclusion, the supplementation of Se and VE could help to improve goat semen quality by increasing the sperm density, sperm motility, the antioxidant enzyme activities, and decreasing the abnormal rate in hot season of Hainan.Finally, Se and VE supplement had good effects on relieving the environment heat stress.     

Prokaryotic Expression and Antigenicity Analysis of Porcine Japenese Encephalitis Virus Envelope Protein Domain Ⅲ

In order to analyze the antigenicity of porcine Japanese encephalitis virus (JEV) E protein domain Ⅲ, which was expressed by pET-28a vector with His-tag and purified through Ni-NTA, the BALB/c mice were immunized with the purified protein.We identified the antigenicity of domain Ⅲ of E protein and the anti-mice and anti-porcine JEV E Ⅲ protein specific antibody titers by SDS-PAGE, Western blotting, indirect ELISA and IFA.SDS-PAGE results showed the expressed target protein existed mainly in the form of inclusion body.Western blotting, ELISA test results showed that the protein had good reactivity with anti-serum.The mice immunized with the purified JEV E Ⅲ protein generated 1×10⁵ anti-JEV E Ⅲ protein specific antibody titers by ELISA, and the porcine immunized with the porcine JEV generated 5.1×10⁴ anti-JEV specific antibody titers.The IFA results showed that JEV E Ⅲ protein anti-serum could identify JEV antigen.The above results showed that the recombinant JEV E Ⅲ had good antigenicity.These results provided important basis for development of diagnostic antigen for JEV.     

Screening of Probiotics Inhibiting E. coli K88 in vitro

The probiotics tested in the experiment were isolated from the intestinal of weaning piglets.The isolated probiotics and E.coli K88 were inoculated into the culture of intestinal porcine epithelial cell-1 (IPEC-1).The activity of lactate dehydrogenase (LDH) in the supernatant was measured after incubating for 2.5 h.At the same time, the probiotics and E.coli K88 were co-cultured in vitro, the number of E.coli K88 was counted and the probiotics which could be resistant to the E.coli K88 were selected 2.5 h later.The results showed that the Lactobacillus casei and Bacillus coagulans could significantly reduce LDH activity (P<0.05), decrease the damage of E.coli K88; Bacillus coagulans could inhibit the growth of E.coli K88.At the same time, Bacillus coagulans could resist high temperature, acid and bile salt.The results showed that Bacillus coagulans strains had great potential as the application of probiotics strains.The methods could be used as a model of screen probiotics which could inhibit the growth of E.coli K88 in vitro.     

Cloning and Sequence Analysis of HA and NA Genes of One Strain of H6N6 Subtype Avian Influenza Virus Isolated from Guizhou Province

To investigate the epidemic situation of H6N6 subtype avian influenza virus (AIV) in Guizhou province,A/duck/Guizhou/013/2014 was isolated from Sansui duck in live poultry market of Guizhou in 2014,the hemagglutinin (HA) and neuraminidase (NA) genes of DK/GZ/14 were subjected to clone and sequence analysis.The results showed that HA gene had the highest nucleotide homologies (97.5%) with the duck-origin H6N6 subtype AIV isolated from Eastern China in 2009,and the strains of HA gene proteolytic cleavage sites was P-Q-I-E-T-R-G,which accordeol with the molecular characteristic of low pathogenic AIV (LPAIV).However,NA gene of A/duck/Guizhou/013/2014 had the highest nucleotide homologies (98.2%) with the duck-origin H6N6 subtype AIV isolated from Fujian in 2007.The phylogenetic tree showed that A/duck/Guizhou/013/2014 and Hunan strains located in the same branch,while three duck-origin H6N6 subtype AIV isolated from Guizhou in 2007 and A/duck/Guizhou/013/2014 located in the different branch for HA and NA genes in genetic evolution,which suggested that A/duck/Guizhou/013/2014 was far with the local H6N6 subtype.The results also clearly indicated that duck-origin H6N6 subtype AIV had genetic diversity in duck population in Guizhou.     

高温胁迫对丹参光合生理指标及叶绿素荧光特性的影响

以丹参为试材,采用土培盆栽法,于第1茬花开前进行高温胁迫处理,研究了高温胁迫对丹参生理指标、光合及叶绿素荧光特性的影响。结果表明:随着高温胁迫时间的延长,丹参叶片超氧化物歧化酶(SOD)活性呈上升趋势,而过氧化氢酶(CAT)活性呈下降趋势;丙二醛(MDA)、游离脯氨酸(Pro)含量均呈上升趋势,且与胁迫时间呈极显著多项式关系。净光合速率(Pn)、气孔导度(Gs)、胞间二氧化碳浓度(Ci)和蒸腾速率(Tr)均呈下降趋势,且各指标均与胁迫时间呈负相关。最大光化学量子产量(Fv/Fm)、PSⅡ潜在光化学活性(Fv/Fo)、有效光化学量子产量(Fv′/Fm′)、光化学淬灭系数(qP)和表观光合电子传递速率(ETR)均呈下降趋势,且各指标均与胁迫时间呈负相关,非光化学淬灭系数(NPQ)呈先上升后下降趋势。研究表明,高温(40℃)对丹参生理指标、光合及叶绿素荧光特性有较大的影响,且各功能指标共同作用,以减轻植株所受伤害。