牦牛病毒性腹泻/粘膜病的防制研究

20世纪80年代以来，我国牦牛群中陆续发现牛病毒性腹泻／粘膜病(BVD／MD)，血清阳性率在30％～42．4％之间，病死率在30％左右，本研究先后从四川、西藏等地牦牛中分离出病毒，并对其进行各种生物学特征鉴定后，表明该病毒与标准毒属同一种，所不同的是四川牦牛病毒株属非致细胞病变型，即属NCP型。但回归本动物能复制出典型病例。目前尚无国产牛粘膜病疫苗用于生产。本研究依据猪瘟病毒与牛病毒性腹泻／粘膜病病毒具有交叉免疫性的原理，用猪瘟弱毒苗对牦牛病毒性腹泻／粘膜病进行预防，试验证明用猪瘟弱毒苗可以预防牛病毒性腹泻／粘膜病，且安全可靠，具有实用价值。     

畜产品安全中存在的问题与分析

阐述了我国畜产品安全现状，强调了其形势的严峻性和紧迫性，分析了当前存在的主要问题，并提出了改进措施和建议。     

甜菜碱对猪不同阶段脂肪和蛋白质代谢的影响

对断奶仔猪、生长猪和肥育猪分别饲喂添加甜菜碱800、1000、1750mg/kg的饲粮，分别进行为期36、62、40d的饲养试验，并进行屠宰取样和分析测定。结果表明甜菜碱使断奶仔猪、生长猪、肥育猪的日增重分别提高8.71%(P<0.05)、13.20%(P<0.01)、13.32%(P<0.01);胴体脂肪率分别降低12.97%(P<0.01)、19.62%(P<0.01)、12.49%(P<0.01)；瘦肉率分别提高4.08%(P<0.05)、7.15%(P<0.05)、3.31%(P<0.05);肝脏中游离肉碱含量分别增加8.98%(P<0.05)、21.36%(P<0.05)、30.18%(P<0.01);血清中生长激素(GH)水平分别增高46.15%(P<0.01)、102.11%(P<0.01)、58.33%(P<0.05)；胰岛素样生长因子-1(IGF-1)水平分别增高38.74%(P<0.05)、44.74%(P<0.01)、47.95%(P<0.01)。生长猪和肥育猪皮下脂肪组织中苹果酸脱氢酶(MDH)的活性分别上升13.60%(P<0.05)和14.59%(P<0.01)，断奶仔猪有提高趋势，但差异不显著(P>0.05)。     

一株枯草芽孢杆菌产抗菌肽培养基筛选及发酵工艺优化的研究

试验旨在对一株产抗菌肽枯草芽孢杆菌BL0006的发酵培养基和发酵条件进行优化,以提高发酵液中抗菌肽的效价。首先通过单因子试验筛选出培养基的最适碳源、氮源与无机盐分别为葡萄糖、蛋白胨和K_2HPO_4;在此基础上采用最陡爬坡路径逼近最大响应区域,再利用Box-Behnken试验设计及响应面分析法得到培养基最佳配方:通过摇瓶单因素试验对发酵条件、温度、接种量、转速进行了优化,优化后的发酵培养基为葡萄糖47.7 g/L、蛋白胨29.0 g/L、K_2HPO_4 3.3 g/L,摇瓶发酵工艺为接种量30 mL/L、初始pH 7.5、摇床转速220 r/min、培养温度37℃。优化后抗菌肽的效价提高到了4.25×10~4 U/mL,是优化前的3.51倍。综合试验结果,本研究筛选的培养基和优化后的发酵工艺可达到高产、降低成本、缩短发酵时间的效果。     

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.     

Acute and Sub-chronic Toxicity Test on Rats of a New Kind of Antidiarrheal Chinese Herbal Medicine Compound Preparation for Livestock

In order to understand the security of a new kind of antidiarrheal Chinese herbal medicine compound preparation for livestock,acute and sub-chronic toxicity test were conducted.Acute toxicity test used the largest drug dose method,20 Wistar rats were orally treated with the Chinese medicine compound preparation.In the sub-chronic toxicity test,80 rats were randomly divided into 4 groups with 20 rats in each group and orally given a dose of 3 000,1 500,750 and 0 mg/(kg·BW）of Chinese medicine compound preparation once a day for 30 days.The general clinical status was observed,rats weight were measured and the dose was adjusted every week during the test,after the test measured blood routine index,biochemistry index,and preceded the gross anatomy observation,weighing each major organs and calculated the viscera coefficient,and proceded main viscera histopathological observation between the high dose group and the control group.The acute toxicity results showed that every rat would be alive gavaged with the lethal dose（LD₅₀）of compound preparation larger 5 g/(kg·BW).The sub-chronic toxicity autopsy showed that except heart,lung,and testicles in individual rats appeared mild bleeding in the high dose group,the other dose group organs found no abnormal change.The haematological index showed except mononuclear cell rate（P<0.05),and hematocrit declined significantly（P<0.05）in the high dose group,all the indexes of the other groups were in the normal range,there was no significant difference from the control group.The test suggested the Chinese medicine compound preparation was no toxicity under the condition of this test according to acute toxicity classification standard of exogenous chemicals by WTO,there was no effect on the growth and development of rats in the sub-chronic toxicity test,and there was no chronic toxicity at least 1 500 mg/kg feeding conditions in short-term repeated application.     

3种乳酸菌制剂对苜蓿和羊草的青贮效果

以苜蓿Medicago stativa和羊草Leymus chinensis为研究对象,通过对其发酵品质和化学成分的研究,探讨Lalsil Dry（LD）、H/MF INOCULANT（H/MF）和青宝II号（FS）3种乳酸菌制剂对羊草和苜蓿青贮的影响。结果表明：羊草直接青贮较易成功,添加LD、FS和质量分数0.01%的H/MF可以显著提高青贮料乳酸生成量（P<0.05）,显著降低丁酸生成量（P<0.05）,获得优质的青贮饲料;苜蓿直接青贮pH值和丁酸含量较高,不能调制优质青贮饲料,在试验中添加3种乳酸菌制剂亦未获得优质的青贮料,但LD处理组和0.002 5% FS处理对青贮料的发酵品质较对照有一定的改善。另外,3种乳酸菌制剂处理青贮料中的中性洗涤纤维和酸性洗涤纤维含量与对照无显著差异（P>0.05）。     

不同咸蛋腌制温度对禽流感毒灭活的影响

经人工接种H9亚型禽流感病毒(AIV)的鸭蛋,分别在12℃、22℃和32℃条件下按常规方法腌制于饱和盐水中,同时以置饱和盐水中的禽流感病毒和未经处理禽流感病毒样品作为对照,通过MDCK细胞培养、间接免疫荧光方法定期进行禽流感病毒毒力测定,结果在12℃、22℃和32℃条件下腌制鸭蛋中的禽流感病毒分别于49天、27天和4天失去毒力,置饱和盐水中的禽流感病毒分别在27天、9天和2天失去毒力,而未经处理的禽流感病毒分别在92天、29天和17天失去毒力;不同试验温度条件下,以荧光RT-PCR检测腌制鸭蛋和对照样品中的禽流感病毒,在100天后仍可检出病毒核酸(Ct<30)。以上检测结果表明,在腌制鸭蛋时于常温下置饱和盐水腌制40天以上的传统咸蛋生产工艺,可使禽流感病毒完全失去毒力,腌制的鲜咸蛋携带或传播禽流感病毒的风险极低或不存在风险。     

高梁11A与3种苏丹草杂交新品系的AFLP分析

利用AFLP分子标记技术,对4个高丹草新品系11A-黑壳苏丹草、11A-白壳苏丹草、11A-棕壳苏丹草、11A-黑壳苏丹章晚熟型与对照品种蒙农青饲1号、蒙农青饲2号、蒙农3号及乐食高丹草的遗传差异性进行比较分析.结果表明:用筛选出的24对适宜引物共扩增出3986个AFLP位点,其中多态性位点3546个,多态性位点比率高达88.96%;各供试材料的遗传距离(GD)变动在0.6905～0.8089之间,以GD值0.74为基准,8个材料聚为3类,第一类为蒙农青饲1号、蒙农青饲2号、蒙农3号、11A-黑壳苏丹草、11A-白壳苏丹草、11A-黑壳苏丹草晚熟型;第二类为乐食;第三类为11A-棕壳苏丹草.