ULTRASONOGRAPHIC EVALUATION OF THE EXTERNAL EAR CANAL AND TYMPANIC MEMBRANE IN DOGS

Ultrasonographic imaging of the canine external ear canal, tympanic membrane, and tympanic bulla was described in five healthy beagle dogs before and after infusion of saline into the ear canal. Saline served as an acoustic window. With this method, the external ear canal, and tympanic bulla were visible in the same imaging plane and the integrity of the tympanic membrane could be evaluated indirectly by confirming an intact tympanic membrane, which appeared at the end of the ear canal as a hyperechoic line with reverberation. Experimentally, perforated tympanic membrane could be evaluated by identifying anechoic saline in the tympanic bulla lumen. The air and fluid-filled tympanic bulla were also visualized. Ultrasonography with saline as an acoustic window appears to be helpful for the evaluation of the external ear canal, tympanic membrane, and tympanic bulla and it may have the potential to be a useful clinical tool in evaluation of integrity of the tympanic membrane.     

基因型鹅副粘病毒NA-1株全基因组的克隆及特性分析

将本实验室分离保存的NA-1株鹅副粘病毒(GPMV)经SPF鸡胚增殖,收集鸡胚尿囊液进行病毒纯化,提取病毒基因组RNA。参考GenBank已收录的GPMVZJ1株基因组序列,设计了8对特异性引物,RT-PCR法分别特异性的扩增出病毒各基因片段,并将各目的基因片段回收纯化,克隆PGEM-T载体,转化大肠杆菌DH5α,小提质粒选取阳性克隆酶切鉴定并进行序列测定,对测定结果进行拼接得到全长cDNA序列(GenBank登录号:DQ659677)。NA-1株核苷酸比新城疫病毒(NDV)核苷酸多6个碱基,位于1644~1645nt之间,符合NDV核苷酸“六碱基原则”。序列分析结果表明,GPMVNA-1株与各地代表株核苷酸同源性81.2%~91.3%。同时根据各毒株F基因开放阅读框前389个核苷酸序列绘制出系统发育进化树,结果表明GPMNNA-1株与SF02和QY97病毒株同属于基因型,不同于基因IX型NDV的国家标准株F48E9和基因型的LaSota传统弱毒株。     

高邮鸭PRL基因内含子1的SNP检测及其与产蛋性状的相关分析

利用PCR—RFLP技术对高邮鸭催乳素（Prolactin，PRL）基因内含子1进行了SNP检测和测序分析，并对该基因与高邮鸭产蛋性状的相关性进行了研究。结果表明：PRL基因内含子1存在两个Dra Ⅰ酶切位点，其中1个位点具有DraⅠ多态性。该位点由于在1326位点发生了T→C的碱基突变，产生了AA、AB和BB3种基因型。基因型BB和等位基因B的频率最高；BB基因型个体30周龄蛋重极显著高于AB基因型个体（P〈0．01）；AB基因型个体的双黄率显著高于BB基因型个体（P〈0．05）；3种基因型间产蛋数、最长连产天数和开产体重无显著差异。     

四倍体高粱与约翰逊草杂交种饲用品质分析

利用四倍体高粱品系"四沈甜"、不育系"四6A"和约翰逊草,配置"四沈甜"×约翰逊草、"四6A"×约翰逊草和"四6A"×"四沈甜"杂交种,分析了杂交种及杂种分离世代的不同生长阶段的杂种优势及饲草品质。结果表明,在孕穗期各供试材料粗蛋白质含量最高,杂交种蛋白质含量达到10%～15%,随着发育期的推进,含量有所下降。抽穗期,粗脂肪含量达到6.1%～7.9%。各材料的粗纤维含量都在30%以上,不同生长阶段差异不显著。单位能量值在抽穗期最高,达到18MJ/kg左右。综合各营养成分含量和单位能量分析,初步认为四倍体高粱与约翰逊草杂交种可以作为饲草应用,抽穗期为刈割和利用的最佳时期。     

黄土高原不同林龄土壤质地和矿质元素差异研究

对黄土高原不同林龄和不同树种条件下土壤的机械组成和矿质元素含量进行了分析研究,结果表明:①成林土壤砂粒、粉粒含量低于中幼林土壤,粘粒含量高于中幼林土壤,且呈显著性差异;②成林土壤Al2O3、Fe2O3、TiO2、MnO2、K2O含量相对于中幼林土壤有不同程度的增加,除MnO2外,差异均为极显著;成林土壤中Al2O3、Fe2O3、TiO2、MnO2在土壤中下层有不同程度的富集,SiO2、CaO含量降低;③矿质元素含量与土壤质地密切相关。     

Isolation and Identification of a Variant Porcine Epidemic Diarrhea Virus and the Immune Efficacy of Its Inactivated Vaccine

This study was aimed to isolate a mutant strain of porcine epidemic diarrhea virus and prepare a PEDV inactivated vaccine with high valence by suspension culture process for immunizing against PEDV effectively in China.200 small intestines and theirs contents of diarrhea piglets died of diarrhea,collected from many large-scale pig farms in China,were detected by RT-PCR and sequenced,a mutant strain of porcine epidemic diarrhea virus was selected and put on the suspension-cultured Vero cells in a 2 L reactor for virus isolation and continuous cell culture,the harvested virus suspension,which was identified and determined TCID₅₀,was inactivated by formaldehyde and mixed with aluminum hydroxide gel adjuvant to prepare PEDV inactivated vaccine.After its physical behavior,stability viscosity,sterility test were checked out,the safety and immune efficacy were studied by immunizing the pregnant pigs and theirs piglets.The results were as follows:86 samples were detected positive in 200 samples,cytopathy occurred after the mutant strain samples screened were passaged to 5th generation,the virus suspension was harvested in 10th generation and identified as a mutant strain of PEDV,named PEDV-GF10 strain.The virus titer of harvested virus suspension was measured up to 1×10^8.0 TCID₅₀/mL after concentrated.After the vaccine was checked out,the sows,40 and 25 days before delivery in experimental groups,were injected into Xuehai acupoint with 4 mL vaccine and the pigs in blank group were free of immunifications.The results showed that there were no obvious differences in the production status of the sows in experimental groups and blank group and the temperature of theirs 3-day-old healthy piglets injected different doses of vaccine,and the vaccine was safe to the sows and piglets.Forty 3-day-old piglets producted by pregnant sows in experimental groups and blank group were randomly selected and taken 4 mL 1×10^8.0TCID₅₀/mL F10 virus culture.The PEDV morbidity of piglets in blank group was 100% after injection and the antibodies were negative;10% piglets in blank group had mild diarrhea symptoms,the protection rate was up to 90%,antibody of passive immunity in piglets lasted for more than 35 days.Virus titer of mutant strain of PEDV-GF10 improved a lot by suspension cell culture,the PEDV-GF10 inactivated vaccine was safe,and could effectively prevent and control the variation strain of PEDV in China.     

Effects of Lactobacillus and Bacillus subtilis Mixed Fermentation Feed on Growth Performance,Slaughter Performance and Intestinal Morphology of Broilers

The purpose of this experiment was to study the usage and proportion of fermented feed in broiler feed.By adding 0,10% and 20% fermented complete formula feed to the broiler feed,and granulating normally,the influence of the usage mode of the fermented feed on the production performance,slaughter performance,intestinal tract and other related indexes of the broiler feed were studied,so as to provide reference for the application of the fermented feed in the broiler feed.180 1-day-old healthy AA male broilers were randomly divided into 3 groups with 6 replicates in each group and 10 chickens in each replicate.The three treatment groups were fed with 0,10% and 20% of the total fermented feed for 42 days.The results showed that:From the whole period (1-42 days old),the 10% fermentation feed substitution group significantly reduced the feed consumption and weight gain ratio (P<0.05),and the average daily weight gain increased (P=0.05).At 42 days old,there was no significant effect of fermented feed on slaughter performance (P>0.05).The ileal crypt depth of broilers in the 20% fermentation feed replacement group was significantly higher than that in the control group and the 10% replacement group (P<0.05).The length of duodenal villi and V/C in 20% and 10% fermentative feed substitution group was significantly higher than that in control group (P<0.05).The results showed that the replacement of 10% fermented feed in broiler feed could reduce the ratio of feed to weight gain,increase the average daily gain,and have a certain impact on intestinal development.     

1株高致病性血凝性脑脊髓炎病毒的分离与鉴定

利用细胞培养方法从临床表现神经症状的死亡仔猪脑组织内分离获得1株病毒,同时对该病毒进行电镜负染观察、昆明种小鼠致病性试验、RT-PCR检测和部分基因测序分析。电镜观察可见细胞培养物中有大量的直径约115 nm的冠状病毒样粒子;接种病毒的小鼠均出现典型的神经症状,死亡率100%;血凝性脑脊髓炎病毒(HEV)特异性引物RT-PCR扩增结果阳性;其S基因序列与GenBank中登录的HEV-67 N相应基因序列的同源性高达99.6%。结果表明,分离的病毒为猪血凝性脑脊髓炎病毒,分离株暂命名为HEV-CC-2007。     

猪附红细胞体可溶性抗原的分析

为了解附红细胞体可溶性抗原的特性,本试验对解离下的猪附红细胞体,经超声波裂解制备粗抗原,再经SephadexG-200分离提纯后,应用对流免疫电泳(CIEP)定位特异性抗原蛋白峰,再进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)及免疫印迹(Westernblot),对猪附红细胞体粗提及纯化的可溶性抗原进行了分析。结果表明,猪附红细胞体可溶性抗原的电泳图谱上有四条蛋白带,其分子量分别为77Ku、62Ku、58Ku、29Ku,其中特异性抗原分子量为58Ku和29Ku。从而为附红细胞体抗原成分的进一步分析及该病的免疫学诊断等研究奠定了基础。     

超声波法和超声波酶法提取灵芝多糖的条件研究

灵芝(Ganoderma lucidum)是我国传统中药,具有滋补强壮,扶正固本的作用[1],至今已有2000多年的使用历史。灵芝子实体多糖是重要的有效成份之一,被广泛用作药品及保健食品的原材料。常用的灵芝多糖提取方法有煎煮法、水蒸馏法等,但这些方法耗时长、提取率低、提取物中多糖含量相