应用RT-PCR技术扩增禽传染性支气管炎病毒S_1基因的研究

选择Ｓ１基因做为目的基因,运用ＲＴ－ＰＣＲ技术分别扩增１２株以非免疫鸡胚繁殖并通过超速离心浓缩的鸡胚尿囊液中的ＩＢＶ,所用引物为ＩＢＶＢｅａｕｄｅｔｔｅ株Ｓ１基因两侧的对应序列,跨幅为１．７２ｋｂ。结果６株ＩＢＶ毒株（ＳＡＩＢ３、Ｆ、Ｄ４１、Ｍ４１、Ｈ５２、Ｈｏｌｔｅ）扩增出了长约１．７ｋｂ的Ｓ１基因片段,而另６株ＩＢＶ毒株虽经４次ＲＴ－ＰＣＲ重复后也显阴性。用ＨａｅⅢ内切酶对６个毒株的ＲＴ－ＰＣＲ产物进行酶切,结果显示Ｍ４１、Ｈ５２、Ｄ４１３个ＩＢＶ毒株的Ｓ１基因包含两个ＨａｅⅢ位点,Ｆ株与Ｈｏｌｔｅ株Ｓ１基因包含１个ＨａｅⅢ位点,而ＳＡＩＢ３株Ｓ１基因则已丢失ＨａｅⅢ位点。实验结果表明：运用ＲＴ－ＰＣＲ技术检测ＩＢＶ时,由于Ｓ１基因的核酸序列具有较高的变异率,因此Ｓ１基因不宜做为目的基因。     

牛轮状病毒重组株的血清学和基因型特性

从母牛接种过Ａ组ＢＲＶ株ＮＣＤＶ－Ｌｉｎｃｏｌｎ（Ｐ１：Ｇ６）苗的两个吡邻牛场分离到两株牛轮状病毒（ＢＲＶ），编号为ＮＳ－１和ＮＳ－２，Ｎｏｒｔｈｅｒｎ－ｂｌｏｔｔｉｎｇ试验表明，ＮＳ－１和ＮＳ－２有相的的Ａ组ＲＮＡ电泳模式，而且全部基因片段同源，体外中和试验表明，ＮＳ－１株病毒能被Ｇ６特异性单克隆抗体（ＭｃＡｓ）和抗ＢＲＶＢ６４１株（Ｐ５：Ｇ６）豚鼠高免血清（ＧＰＡＳ）中和，但不被Ｇ１０特异性Ｍ     

京—1株鸡马立克氏病强毒对BWEL—SPF鸡群的感染试验

用京－１株鸡马立克氏病（ＭＤ）强毒感染１￣１０个家系１日龄的ＢＷＥＬ－ＳＰＦ鸡,观察比较临床症状和剖检病变及病理切片,以此评估该ＢＷＥＬ－ＳＰＦ鸡群每个家系鸡对京－１株ＭＤ强毒的敏感性。结果,１,５,８,９号家系的ＢＷＥＬ－ＳＰＦ鸡对京－１株ＭＤ强毒最敏感;３,４,５,６,１０号家庭系次之,２、７号家系敏感性相对较差。     

牛传染性鼻气管炎病毒tk基因的克隆

根据牛传染性鼻气管炎病毒ＬＡ株的物理图谱及Ｃｏｐｅｒ株和ＬＡ标ｔｋ基因在基因组中的定位，将ＬＡ株ＤＮＡ ＨｉｎｄⅢＡ片段中的ＳａｌＩ－ＳａｌＩ亚片段、ＢｇｌⅢ－ＳａｌＩ双酶切亚片段分别克隆到载体质粒ｐＢｌｕｅｓｃｒｉｐｔＳＫ中，筛选出３个重组质粒ｐ＾ｔｋ－１，Ｐｔｋ－２和Ｐｔｋ－３，其外源片段大小分别为２．７ｋｂ，１．１ｋｂ和１．６ｋｂ。经酶切分析及同源核酸探针杂交试验表明，２．７ｋｂ ＳａｌＩ－     

应用RT—PCR技术扩增禽传染性支气管炎病毒S1基因的研究

选择Ｓ１基因做为目的基因,运用ＲＴ－ＰＣＲ技术分别扩增１２株以非免疫鸡胚繁殖并通过超速离心浓缩的鸡胚尿囊液中的ＩＢＶ,所用引物为ＩＢＶ Ｂｅａｕｄｅｔｔｅ株Ｓ１基因两侧的对应序列,跨幅为１．７２ｋｂ。结果６株ＩＢＶ毒株（ＳＡＩＢ３、Ｆ、Ｄ４１、Ｈ５２、Ｈｏｌｔｅ）扩增出了长约１．７ｋｂ的Ｓ１基因片段,而另６株ＩＢＶ毒株虽经４次ＲＴ－ＰＣＲ重复后也显阴性。用ＨａｅⅢ内切酶对６个毒株的ＲＴ－ＰＣＲ产物     

传染性支气管炎病毒类M_(41)地方分离株S_1基因克隆及高变区序列分析

传染性支气管炎病毒（ＩＢＶ）分离株ＱＤ经ＲＴ－ＰＣＲ扩增出约１．７ｋｂＳ１糖蛋白基因ｃＤＮＡ，修饰后插入ｐＵＣ１８ＳｍａｌⅠ／ＥｃｏＲＩ位点构成质粒ｐＵＣＱＤＳ１。对克隆的Ｓ１基因５′端高变区序列测定显示，起始密码上游６０碱基和下游３８０碱基中与参考株Ｍ４１Ｓ１基因序列仅有１个碱基的差异，表明ＱＤ为一类Ｍ４１分离株。     

鸡新城疫——减蛋综合征二联油佐剂疫苗的研制

应用鸡新城疫（ＮＤ）ＬａＳｏｔａ株和减蛋综合征（ＥＤＳ－７６）ＬＡ３株,分别接种鸡胚和鸭胚增殖病毒,血凝价分别达≥１：１２８０和≥１：１００００时,收取病毒,经福尔马林灭活,按一定比例加入吐温－８０、油佐剂制成ＮＤ－ＥＤＳ－７６二联疫苗,经无菌检验、安全性试验、剂量选择、保护性试验、免疫效果和免疫期测定及保存期观察,结果表明：该二联苗安全性好,于蛋鸡开产前１月皮下注射１ｍＬ／只,２５ｄ后ＮＤ和ＥＤＳ－７６抗体可先后达到高峰,免疫期１年。保存期４℃为１年,２０℃为３个月。     

鸡传染性支气管腺胃病变型毒株免疫原基因的分离鉴定

以腺胃病型变型鸡传染性支气管炎病毒分离株Ｈ９５提取的ＲＮＡ为模板,按ＩＢＶ Ｂｅａｕｄｅｔｔｅ株Ｓ１基因两侧对应序列,设计一对２７ｂｐ引物,引物间跨幅为１．７Ｋｂ。用反转录－聚梧酶链反应获得与预期大小一致的核酸片段。ＲＴ－ＰＣＲ产物凝胶电泳后,Ｓｏｕｔｈｅｒｎ印迹,经Ｓ１探针检测呈阳性,表明获得分离株Ｈ９５Ｓ１基因。     

鸡传染性法氏囊病病毒野毒株VP2基因高可变区酶切位点分析

参照澳大利亚鸡传染性法氏囊病病毒（ Ｉ Ｂ Ｄ Ｖ）００２７３ 毒株基因组序列设计的 １ 对引物,位于 Ｖ Ｐ２ 高可变区两端,通过 Ｒ Ｔ Ｐ Ｃ Ｒ,扩增了 ５ 个 Ｉ Ｂ Ｄ Ｖ 山东分离株 Ｖ Ｐ２ 基因的 ６０７～１ ０８０ 位核苷酸片段（ａａ２０３～３０６）。 Ｐ Ｃ Ｒ 产物经纯化后,分别用 Ｄｒａ Ⅰ、 Ｓａｃ Ⅰ、 Ｓｓｐ Ⅰ、 Ｐｓｔ Ⅰ和 Ｓａｕ３ Ａ Ｉ共 ５ 种限制性内切酶（ Ｒ Ｅ）进行消化处理,结果 ５ 个分离株均为 Ｄｒａ Ⅰ（－ ）、 Ｓａｃ Ⅰ（－ ）和 Ｓａｕ ３ Ａ Ｉ（＋ ）, Ｌ Ｃ２ 和 Ｔ Ａ 株为 Ｓｓｐ Ⅰ（＋ ）, Ｌ Ｃ１ 和 Ｊ Ｎ 株为 Ｓｓｐ Ⅰ（－ ）, Ｌ Ｃ１ 和 Ｌ Ｃ２ 株为 Ｐｓｔ Ⅰ（＋ ）, Ｊ Ｎ、 Ｌ Ｌ 和 Ｔ Ａ 株为 Ｐｓｔ Ⅰ（－ ）;５ 个分离株的酶切图谱与美国变异株迥异,而 Ｔ Ａ 株与日本超强毒株 ９０１１ 相类似。５ 个分离株与现用疫苗株对比,至少在 Ｖ Ｐ２ 可变区内存在着一定的差异,这可能是 Ｉ Ｂ Ｄ Ｖ 接种免疫鸡群仍然暴发 Ｉ Ｂ Ｄ 的原因之一。     

传染性支气管炎病毒的分离提纯及蛋白电泳分析

通过ＳＰＦ鸡胚接种分别对传染性支气管炎病毒的２ 个标准毒株Ｍ４１、ＩＢＶ－ Ｔ 株，４ 株山东分离株及１ 株北京分离株进行扩增繁殖；利用差速离心法、蔗糖不连续密度梯度离心法进行提纯；用ＳＤＳ－ 聚丙烯酰胺凝胶电泳法进行蛋白分析。结果表明：各毒株都含有四种结构蛋白，且各毒株结构蛋白分子量基本相似，其中仅Ｍ 和Ｓ蛋白略有区别     

白术、微米白术和白术多糖对断奶仔猪生长性能和肠道形态及微生态区系的影响

试验选用96头平均体重14.82 kg左右的杜×长×大断奶仔猪,随机分成4组,每组3栏,每栏8头(公母各半)。对照组饲喂基础日粮,试验1、2、3组分别添加1%80目白术、0.2%白术多糖和1%微米白术。试验期30 d。结果表明:在生长性能方面,与对照组相比,1%微米白术添加组可显著提高日增重(P0.05)、降低饲料增重比和腹泻率,而且效果优于1%80目白术组和0.2%白术多糖组,在肠道形态和肠道微生态区系方面,与对照组相比,日粮添加1%80目白术、0.2%白术多糖、1%微米白术均可不同程度的提高十二指肠和空肠的绒毛高度,加深十二指肠和空肠的隐窝深度,并且增加肠道微生态区系的多样性,其中以1%微米白术添加组的效果最佳。     

Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia

Sissay, M.M., Uggla, A. and Waller, P.J., XXXX. Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia. Tropical Animal Health and Production, XXXX. A 2-year abattoir survey was carried out to determine the prevalence, abundance and seasonal incidence of gastro-intestinal (GI) nematodes and trematodes (flukes) of sheep and goats in the semi-arid zone of eastern Ethiopia. During May 2003 to April 2005, viscera including liver, lungs and GI tracts were collected from 655 sheep and 632 goats slaughtered at 4 abattoirs located in the towns of Haramaya, Harar, Dire Dawa and Jijiga in eastern Ethiopia. All animals were raised in the farming areas located within the community boundaries for each town. Collected materials were transported within 24 h to the parasitology laboratory of Haramaya University for immediate processing. Thirteen species belonging to 9 genera of GI nematodes (Haemonchus contortus, Trichostrongylus axei, T. colubriformis, T. vitrinus, Nematodirus filicollis, N. spathiger, Oesophagostomum columbianum, O. venulosum, Strongyloides papillosus, Bunostomum trigonocephalum, Trichuris ovis, Cooperia curticei and Chabertia ovina), and 4 species belonging to 3 genera of trematodes (Fasciola hepatica, F. gigantica, Paramphistomum {Calicohoron} microbothrium and Dicrocoelium dendriticum) were recorded in both sheep and goats. All animals in this investigation were infected with multiple species to varying degrees. The mean burdens of adult nematodes were generally moderate in both sheep and goats and showed patterns of seasonal abundance that corresponded with the bi-modal annual rainfall pattern, with highest burdens around the middle of the rainy season. In both sheep and goats there were significant differences in the mean worm burdens and abundance of the different nematode species between the four geographic locations, with worm burdens in the Haramaya and Harar areas greater than those observed in the Dire Dawa and Jijiga locations. Similar seasonal variations were also observed in the prevalence of flukes. But there were no significant differences in the prevalence of each fluke species between the four locations. Overall, the results showed that Haemonchus, Trichostrongylus, Nematodirus, Oesophagostomum, Fasciola and Paramphistomum species were the most abundant helminth parasites of sheep and goats in eastern Ethiopia.     

Feline intracranial neoplasia: retrospective review of 160 cases (1985-2001)

The purpose of this study was to determine the frequency of different tumor types within a large cohort of cats with intracranial neoplasia and to attempt to correlate signalment, tumor size and location, and survival time for each tumor. Medical records of 160 cats with confirmed intracranial neoplasia evaluated between 1985 and 2001 were reviewed. Parameters evaluated included age, sex, breed, FeLV/FIV status, clinical signs, duration of signs, number of tumors, tumor location(s), imaging results, treatment, survival times, and histopathologic diagnosis. Most of the cats were older (11.3 +/- 3.8 years). Primary tumors accounted for 70.6% of cases. Metastasis and direct extension of secondary tumors accounted for only 5.6 and 3.8% of cases, respectively. Twelve cats (7.5%) had 2 or more discrete tumors of the same type, whereas 16 cats (10.0%) had 2 different types of intracranial tumors. The most common tumor types were meningioma (n = 93, 58.1%), lymphoma (n = 23, 14.4%), pituitary tumors (n = 14, 8.8%), and gliomas (n = 12, 7.5%). The most common neurological signs were altered consciousness (n = 42, 26.2%), circling (n = 36, 22.5%), and seizures (n = 36, 22.5%). Cats without specific neurological signs were common (n = 34, 21.2%). The tumor was considered an incidental finding in 30 (18.8%) cats. In addition to expected relationships (eg, meninges and meningioma, pituitary and pituitary tumors), we found that lesion location was predictive of tumor type with diffuse cerebral or brainstem involvement predictive of lymphoma and third ventricle involvement predictive of meningioma.     

Y‐chromosomal variation of local goat breeds of Turkey close to the domestication centre

Genetic variations in chromosome Y are enabling researchers to identify paternal lineages, which are informative for introgressions and migrations. In this study, the male‐specific region markers, sex‐determining region‐Y (SRY), amelogenin (AMELY) and zinc finger (ZFY) were analysed in seven Turkish native goat breeds, Angora, Kilis, Hair, Honaml?, Norduz, Gürcü and Abaza. A SNP in the ZFY gene defined a new haplotype Y2C. All domestic haplogroups originate from Capra aegagrus, while the finding of Y1A, Y1B, Y2A and Y2C in 32, 4, 126 and 2 Turkish domestic goats, respectively, appears to indicate a predomestic origin of the major haplotypes. The occurrence of four haplotypes in the Hair goat and, in contrast, a frequency of 96% of Y1A in the Kilis breed illustrate that Y‐chromosomal variants have a more breed‐dependent distribution than mitochondrial or autosomal DNA. This probably reflects male founder effects, but a role in adaptation cannot be excluded.     

A brief history of the prevention of infectious diseases by immunisations

Infectious diseases have always been a terrible scourge for humans. The appearance of these plagues, as they were called without distinction, was generally connected to various conditions: asters, climatic changes or religious reasons. The concept of contagious, and then infectious, diseases came slowly. Variolation, i.e. transmission of ‘virulent’ matter to induce a natural disease and the immunity against it, was brought from Constantinople to England by Lady Montague, in 1721. This ‘variolation’ technique was also often performed in veterinary medicine against diseases like sheep-pox or pleuropneumonia. As ‘vaccination’ is the term generally accepted for ‘immunisation’, variolation can be the word designating such a technique. The second period of the history of immunisation began, in 1880, with the studies of Pasteur and his collaborators. A great number of bacterial vaccines were developed: dead, live but attenuated or only parts of pathogens. The viruses were produced in animals, then in eggs and at last, in tissue cultures. Second generation vaccines appeared with genetic engineering: recombinant vaccines, vector vaccines, nucleic acids vaccines, and markers vaccines, among others. These novel technologies can permit the development of new ones and improve the quality of the vaccines already existing.     

Micronutrient Content of Certain Tropical Conventional and Unconventional Feed Resources of Southern India

The ash, silica and certain important micronutrients were estimated in conventional and unconventional feed and fodder resources available in Southern India. Commonly used dry roughages, such as paddy straw, ragi straw, maize kadbi/stalk, jowar kadbi/stalk, bajra stalk and wheat straw, were high in ash (9.9% +/- 0.77%) and silica (6.4% +/- 0.65%) and low in most of the other micronutrients, except iron, with paddy straw containing most silica (>9%). Cultivated non-leguminous (maize, jowar) and improved green crops (hybrid napier, guinea, green panic, NB-21, CO-1) were also high in ash (10.5% +/- 0.60% and 12.5% +/- 0.51%) but were moderate sources of P, Mg and Cu and good sources of Zn (98 +/- 13.8 ppm and 55 +/- 6.7 ppm). Leguminous green fodders (stylosanthus, lucerne, cow pea, soyabean) were excellent sources of Ca (1.9% +/- 0.16%), Mg (0.40% +/- 0.05%), Cu (30 +/- 5.2 ppm), Zn (121 +/- 14.7 ppm) and Fe (1234 +/- 166 ppm) and moderate sources of P. Mixed local grasses and weeds were high in silica (6.9% +/- 1.00%) but were good sources of Cu, Zn and Fe. Cereal grains (maize, wheat, rice, ragi) were low in ash (2.9% +/- 0.33%) and were relatively poor sources of Ca (0.22% +/- 0.03%), Mg (0.19% +/- 0.03%) and Cu (13 +/- 3.1 ppm). Pulses were low to medium sources of most minerals and good sources of Fe (1230 +/- 293 ppm). Oil seed cake/extractions (groundnut cake, cotton seed cake, soyabean meal, sunflower cake, safflower cake) and cereal by-products (rice polish, rice bran, wheat bran) were excellent sources of P (1.1% +/- 0.47% and 2.3% +/- 0.19%) and good sources of Zn (65 +/- 3.9 ppm and 66 +/- 10.7 ppm) and Fe (938 +/- 130 ppm and 662 +/- 126 ppm). Among the unconventional feeds screened, orange peel, sunflower heads, meat meal, rubber seed cake, spirulina algae and sea weeds contained plentiful Ca, Zn and Fe: tree leaves/top feeds (mulberry, erythrina, glyricidia, banana, subabul, groundnut haulms) were excellent sources of Ca (1.5% +/- 0.13%), Zn (120 +/- 22.9 ppm) and Fe (1033 +/- 133 ppm) but relatively poor sources of P. Soyabean husk, cocoa seed husk, rubber seed cake and meat meal were moderate to good sources of P (1.0% and 0.33%). The high Zn and Fe values of most feeds/fodders were probably due to soil contamination. This account of the micronutrient content of feed/fodder resources should help in strategic supplementation intended to alleviate local deficiencies.     

Bovine Mastitis in Selected Areas of Southern Ethiopia

A study on bovine mastitis, designed to determine the causal agents, prevalence of infection and impact of risk factors in three cattle breeds, was conducted in selected areas of southern Ethiopia. A total of 307 lactating and non-lactating cows, of which 162 were indigenous Zebu, 85 Jersey and 60 Holstein-Friesian, were examined by clinical examination and the California mastitis (CMT) test. Of these, 40.4% were positive by CMT and bacteriology for clinical or subclinical mastitis, with prevalence rates of 37.1% and 62.9%, respectively. Out of 1133 quarters examined, 212 (18.7%) were found to be infected, 83 (39.2%) clinically and 129 (60.8%) subclinically. The prevalence of mastitis was significantly higher in Holstein-Friesian than in indigenous Zebu, in non-lactating cows than in lactating cows, in the early lactation stage than in the mid-lactation stage, in cows with lesions and/or tick infestation on skin of udder and/or teats than in cows without this factor, and in the wet season than in the dry season. Mastitis increased with parity number (R = 0.9). Of 248 CMT and clinically positive udder quarter samples analysed microbiologically, 212 were culturally positive for known mastitis pathogens and 36 were negative. Of the 199 positive samples, Staphylococcus accounted for 39.2%, Streptococcus for 23.6%, coliforms for 14.1%, Micrococcus and Bacillus species for 8.0% each and Actinomyces or Arcanobacterium (Corynebacterium) for 7.0%. It was concluded that there was a high prevalence of clinical and subclinical mastitis, mainly caused by Staphylococcus aureus, Streptococcus agalactiae and Escherichia coli, in this study area.     

不同日粮氮水平对山羊氮代谢和微生物蛋白质合成的影响

本试验旨在探讨不同日粮氮水平对内蒙古白绒山羊氮代谢和微生物蛋白质合成(MCP)的影响,以达到提高氮利用率,减少环境污染和饲料资源的浪费。试验选用9只体况良好、体重为(43.83±2.95)kg,装有瘤胃瘘管的内蒙古白绒山羊,按体重随机分为3组,每组3只。日粮分为低氮7.5%、中氮10.5%、高氮13.5%3个氮水平。试验使用常规试验方法分析测定粪尿血液等生化指标,通过全收尿法和尿嘌呤衍生物法估测MCP。结果表明:随着氮水平增加,氨氮(NH3-N)浓度、氮摄入量、尿氮排出、尿中尿素氮(UUN)和MCP显著增加(P0.05),干物质采食量(DMI)显著降低(P0.01);沉积氮占总摄入氮的比例在中氮组最高,MCP在高氮组最高。综上所述,适当降低日粮氮水平可减少粪尿氮排放,提高反刍动物氮利用率。     

Microsporidial keratopathy in two dogs

A microsporidial keratopathy is described in two dogs. Both dogs presented with a unilateral stromal keratopathy characterized by multifocal coalescing opacities, and the diagnosis was made on histopathologic examination of keratectomy specimens. Transmission electron microscopy (TEM) on formalin‐fixed, paraffin‐embedded corneal tissue was performed in one dog, and the morphologic features were consistent with Nosema species infection. Both dogs were initially diagnosed and treated by superficial keratectomy. One dog received additional antifungal medication and underwent a penetrating keratoplasty following local recurrence two years later. No other systemic lesions attributable to the microsporidial infection were identified clinically. The clinical and diagnostic pathology findings, treatment, and follow‐up are discussed.