奶牛主要植物性蛋白质饲料蛋白质和纤维物质瘤胃降解率的研究

选用3头安装永久性瘤胃瘘管的健康荷斯坦泌乳奶牛为试验动物,选取8种北方地区奶牛常用蛋白质饲料原料为试验样品,每个样品设3个重复,用尼龙袋法分别测定各个样品的粗蛋白质（CP）、中性洗涤纤维（NDF）和酸性洗涤纤维（ADF）在瘤胃内的动态降解率和有效降解率。结果表明,①所选8种蛋白质饲料原料中葵花粕的粗蛋白质（CP）有效降...     

Comparison of heat tolerance of feedlot heifers of different breeds

Heat stress in cattle causes decreases in feed intake and feed efficiency; in extreme cases, it can cause death. These losses amount to millions of dollars each year. A study was designed to determine severity of heat stress among four breeds of cattle. Throughout two summers, 256 feedlot heifers of four different breeds were observed. Respiration rates, panting scores, and surface temperatures were taken twice each day on 10 animals/breed for several weeks during the summers of 2002 and 2003. Twenty-four-hour behavior measurements were recorded for four heat-stress and four thermoneutral days. Results showed during the afternoon, Angus cattle (black) had the highest respiration rates, panting scores, and surface temperatures, followed by the MARC III (dark red), Gelbvieh (tan), and Charolais (white). Behavior data showed that heat stress increased drinking and standing behavior, and decreased eating, lying, and agonistic behaviors, and that dark-hided cattle adjusted their behavior more than light-hided cattle. Overall, it was found that breed of cattle with dark-hides were more affected by temperature changes and at peak temperatures than breeds of cattle with light-hides.     

鸡蛋粉在猪生产中的应用研究进展

鸡蛋粉是一种良好的蛋白质资源,富含免疫活性物质,在减少动物腹泻、增强机体免疫力方面具有重要功效。本文综述了鸡蛋粉的生产工艺、营养价值以及鸡蛋粉和特异性卵黄抗体在猪生产中的应用现状。     

牛至油和肉桂醛添加组合对奶牛瘤胃微生物蛋白产量与养分消化率的影响

本试验旨在研究牛至油(Oo)与肉桂醛(CA)的不同添加组合对奶牛瘤胃微生物蛋白产量与养分消化率的影响。选取年龄、体重、胎次、产奶量、乳成分相近及泌乳期为(90±15)d的荷斯坦奶牛40头,随机分为10个组,每组4头。对照组(C)饲喂基础日粮,试验组饲喂不同添加水平的Oo和CA,Oo 3个梯度分别为11.5(L)、13(M)、14.5(H)g/(d·头);CA 3个梯度分别为15(L)、18(M)、21(H)g/(d·头),共组成9个不同的添加量组合,分别为LL、ML、HL、LM、MM、HM、LH、MH、HH(第1个字母为Oo添加水平,第2个字母为CA添加水平)。预试期15 d,正试期60 d。结果表明:试验组奶牛的瘤胃微生物蛋白(MCP)产量均有提高趋势,其中LH组效果最好,瘤胃MCP产量比C组提高了12.82%(P<0.05);试验组奶牛在养分表观消化率方面有提高趋势,其中以LH组最优,干物质与粗蛋白质的消化率高于C组(P<0.05),中性洗涤纤维与酸性洗涤纤维的消化率高于C组(P<0.01)。本试验条件下,综合考虑瘤胃MCP产量以及各主要养分的表观消化率,以添加Oo 11.5 g/(d·头)、CA 21 g/(d·头)为最佳。     

急性热应激对山羊血液生化指标及血淋巴细胞热休克蛋白70家族基因表达的影响

旨在研究急性热应激对山羊抗氧化能力、免疫功能和血淋巴细胞热休克蛋白70(HSP70)家族基因表达的影响。本试验选取5只健康、体况接近的(12±0.5)月龄波尔山羊×关中奶山羊杂交F1母羊,饲养于环控舱内(温度维持20℃,相对湿度60%),适应5d。第6天利用环控舱对5只试验羊进行38℃急性热应激处理12h,采集热应激前(0h,20℃)和热应激后2、4、8和12h试验羊血样。分别利用比色法测定血清抗氧化指标(总抗氧化能力、超氧化物歧化酶、谷胱甘肽过氧化物酶活性和丙二醛含量),ELISA法测定血清免疫指标(免疫球蛋白和细胞因子含量),实时荧光定量PCR法测定血淋巴细胞HSP70家族基因(HSPA1A、HSPA6和HSPA8)mRNA的表达量。结果显示:1)热应激时间对血清抗氧化指标有显著影响。与热应激前0h相比,血清T-AOC(P<0.05)、SOD(P<0.05)和GSH-Px(P<0.01)活性均在热应激8h后显著下降,MDA含量在热应激4h后显著增加(P<0.05)。2)热应激时间对血清免疫指标有显著影响。与热应激前0h相比,血清TNF-α、IL-1β、IFN-γ和IL-2含量分别在热应激4、8、8和4h后显著增加(P<0.05);IL-4(P<0.01)、IgG(P<0.01)、IgM(P<0.01)和IgA(P<0.05)含量分别在热应激12、4、4和4h后显著下降。3)热应激时间显著提高血淋巴细胞中HSP70家族基因(HSPA1A、HSPA6和HSPA8)的表达量。HSPA1AmRNA表达量呈先升高后下降的趋势,在热应激4h时达到峰值,各检测时间点均显著高于应激前水平(P<0.01);HSPA6mRNA表达量在热应激2h时显著升高(P<0.01),4h后恢复到应激前水平(P>0.05);HSPA8mRNA表达量在热应激4(P<0.05)、8(P<0.01)、12h(P<0.01)时显著高于应激前水平。在本试验条件下,38℃急性热应激能够抑制山羊的免疫和抗氧化功能;提高血淋巴细胞HSPA1A、HSPA6和HSPA8基因的表达量,其中HSPA1A对热应激温度和时间更敏感,可作为山羊热应激早期的分子标志物。     

Prokaryotic Expression and Polyclonal Antibody Preparation of African Swine Fever Virus Georgia 2007/1 Strain CD2v Protein

The study was aimed to express the EP402R gene of African swine fever virus (ASFV) Georgia 2007/1 strain via prokaryotic expression system,obtain the recombinant CD2v protein,and prepare polyclonal antibodies against the purified recombinant CD2v protein.After codon optimization,ASFV EP402R full-length gene was linked into pET-28a(+) expression vector to construct prokaryotic recombinant expression plasmid.After induction by 1 mmol/L IPTG at 16 ℃ for 12 h,the recombinant protein was identified by SDS-PAGE and Western blotting.The purified recombinant CD2v protein was used as immunogen to prepare mouse anti-CD2v polyclonal antibodies.The antibody titer was measured by indirect ELISA and the specificity was further analyzed by indirect immunofluorescence assay (IFA) and Western blotting.The results showed that ASFV EP402R gene was successfully cloned into pET-28a(+),and pET-28a-EP402R was obtained.The recombinant plasmid was transformed into E.coli BL21(DE3) for expression,the recombinant protein was expressed mainly in the form of inclusion bodies,with molecular mass at about 47 ku,while some of the recombinant protein could also exist in a soluble form.Western blotting results showed that the purified protein had good immunoreactivity.The indirect ELISA result showed that the polyclonal antibodies had a high titer of 1:512 000,IFA and Western blotting results indicated that it could specifically recognize recombinant CD2v protein.These results confirmed the recombinant CD2v protein expressed via prokaryotic system had good immunogenicity,and the prepared polyclonal antibodies had high titer and specificity.This research provided technical support for further study of ASFV EP402R biological function,as well as its gene-deletion based vaccine development.     

小反刍兽疫病毒双抗原夹心时间分辨荧光免疫测定方法的建立

通过优化大肠埃希菌密码子、优化表达条件等方法,在大肠埃希菌表达系统中成功获得了可溶性的N蛋白与NH融合蛋白。进一步基于纯化的可溶性N蛋白与NH融合蛋白建立了小反刍兽疫病毒双抗原夹心时间分辨荧光免疫测定方法。该方法敏感性高,特异性强,能够特异性的检测羊血清中的小反刍兽疫病毒抗体,与其他相关疫病病原无交叉反应;重复性试验组内与组间变异系数均小于10%。对292份临床血清样品进行检测,与法国IDVET竞争ELISA试剂盒比较,阳性样品符合率为92.47%,阴性样品符合率为97.26%,总的符合率为94.86%。该方法与传统的ELISA方法相比,具有敏感性、特异性相当,操作简单、快速,具有较高的推广应用价值。     

紫花苜蓿PGIP同源基因克隆与序列核苷酸多态性分析

多聚半乳糖醛酸酶抑制蛋白(PGIP)是植物重要的防卫蛋白之一,了解PGIP基因的核苷酸序列是对其进行分子遗传学与分子生物学研究的前提与基础。本研究采用同源克隆法,从16个紫花苜蓿(Medicago sativa)品种的集群DNA中克隆了7个苜蓿PGIP基因的部分基因组编码序列,分别对应于蒺藜苜蓿(M. truncatula)7个不同的PGIP基因,并命名为MsPGIP5、MsPGIP6、MsPGIP7、MsPGIP8、MsPGIP9、MsPGIP10和MsPGIP11。其中,MsPGIP5、MsPGIP6、MsPGIP8、MsPGIP9和MsPGIP11的单核苷酸多态性(Single Nucleotide Polymorphism,SNP)位点间的平均距离分别为20、74、34、423和21 bp,差异较大。对MsPGIP5、MsPGIP6、MsPGIP8和MsPGIP11进行荧光标记PCR单链构象多态性(FPCR-SSCP)分析,结果表明,其等位基因数目与PCR片段长度的比值分别为20、56、22和19 bp,变化趋势与这4个基因SNP位点间的平均距离基本一致。比较而言,在发现的7个苜蓿PGIP基因中,MsPGIP9序列高度保守,MsPGIP5和MsPGIP11的变异较大,而MsPGIP6与MsPGIP8的变异中等。     

内质网应激偶联炎性反应研究进展

内质网是真核细胞蛋白合成和折叠的主要细胞器,当内质网内蛋白合成或折叠负担增加,引起未折叠或错误折叠蛋白增多时,可激活内质网几条特定信号通路,启动未折叠蛋白反应,这对维持细胞稳态有重要意义。越来越多研究表明,多种炎性反应疾病与内质网应激有密切联系。一方面,内质网应激引起的未折叠蛋白反应可以诱发或者抑制炎症,另一方面,炎性反应也能影响蛋白折叠,从而促进或缓解内质网应激。2型糖尿病、肿瘤和动脉粥样硬化等多种重大疾病的病理机制都涉及内质网应激与炎性反应的相互作用,对该问题的深入研究不仅能加深人们对这些疾病发病机制的理解,也有助于相关药物的研发。     

北方须鳅幼鱼的饲料蛋白质需求量

为确定北方须鳅幼鱼对饲料蛋白质的需求量,进行了为期60 d的生长试验。以鱼粉、发酵豆粕和酪蛋白作为蛋白质源,设计了蛋白质水平分别为22.85%、31.09%、40.72%、47.21%和54.08%的5种等脂等能试验饲料。每种饲料设3个重复,每个重复放养平均体重为(0.77±0.02)g的北方须鳅幼鱼40尾,以重复为单位养殖于水体为80 cm×50 cm×20 cm的水槽中。结果表明:饲料蛋白质水平为22.85%~40.72%时,随着饲料蛋白质水平的升高,增重率和特定生长率显著升高(P0.05);再继续升高饲料蛋白质水平,增重率和特定生长率反而显著降低(P0.05)。在饲料蛋白质水平为22.85%~40.72%时,饲料转化效率和存活率随着饲料蛋白质水平的升高呈升高趋势;当饲料蛋白质水平为40.72%~54.08%时,饲料转化率和存活率随着饲料蛋白质水平的升高呈下降趋势。在饲料蛋白质水平为40.72%时,试验鱼获得最高的增重率、特定生长率、饲料转化效率和存活率。饲料蛋白质水平对北方须鳅幼鱼的肥满度、脏体比、肝体比以及全鱼粗蛋白质和水分含量均未产生显著影响(P0.05)。饲料蛋白质水平高于40.72%后,全鱼粗脂肪含量显著降低(P0.05),并且饲料蛋白质水平为31.09%~54.08%时全鱼粗脂肪含量随着饲料蛋白质水平的升高而下降。由增重率与饲料蛋白质水平建立二次回归方程,得出北方须鳅幼鱼达到最大生长速度时的饲料蛋白质需求量为41.57%。