抗猪传染性胃肠炎病毒杂交瘤细胞构建及单克隆抗体制备

TGEV-PL株在PK15细胞上增殖,经浓缩纯化获得TGEV抗原,建立了间接ELISA检测方法.应用杂交瘤技术获得3株分泌抗猪传染性胃肠炎病毒(TGEV)单克隆抗体(McAb)杂交瘤细胞.经检测其分泌的抗体亚类为IgG3;杂交瘤细胞染色体数为88;间接ELISA检测细胞培养上清液效价为1∶256,腹水抗体效价达1∶6×104,与6株毒(菌)株的抗原之间无交叉反应.经阻断试验证实,其分泌的McAb能识别抗原是TGEV所特有的抗原决定基.     

己烯雌酚单克隆抗体的制备及鉴定

用人工合成的己烯雌酚-牛血清白蛋白(DES-BSA)作为抗原免疫Balb/c小鼠,利用杂交瘤技术建立了1株稳定分泌己烯雌酚单克隆抗体的杂交瘤细胞1F5,经鉴定:杂交瘤细胞染色体数为99.24±2.5,其分泌的抗体亚型为IgG2α。该细胞株体外传代和冻存复苏后抗体分泌稳定。间接ELISA测定培养上清效价为1∶640,诱生腹水效价3.2×106。ciELISA检测显示其DES 50%抑制质量浓度(IC50)为24μg/L,与水产养殖中常见禁用激素和抗菌药物的交叉反应小,表明该单抗具有较好的敏感性和特异性。     

马绒毛膜促性腺激素单克隆抗体的制备及鉴定

本研究采用马绒毛膜促性腺激素(equine chorionic gonadotropin,eCG)纯品免疫BALB/c小鼠,取其脾细胞与同系小鼠骨髓瘤细胞SP2/0按1∶10融合,间接酶联免疫吸附法(ELISA)筛选阳性克隆和有限稀释法克隆,扩大培养后于小鼠腹腔制备腹水,共得到8株能稳定分泌单克隆抗体的杂交瘤细胞株,采用间接ELISA测定抗体效价,效价均达10^-5以上。用获得的杂交瘤细胞株制备单克隆抗体,为eCG分泌水平的ELISA检测方法的建立确立了条件。     

无色孔雀石绿单克隆抗体的制备与鉴定

用人工合成的无色孔雀石绿-牛血清白蛋白(LMG-BSA)作为抗原免疫BALB/c小鼠,并利用杂交瘤技术建立了1株稳定分泌无色孔雀石绿(LMG)单克隆抗体的杂交瘤细胞2E6.经鉴定表明,杂交瘤细胞染色体数为(99.12±3.60),间接竞争ELISA(ciELISA)显示LMG 50%抑制质量浓度为20.39μg/L,诱生小鼠腹水的抗体效价达2.3×105.与水产养殖中常见的禁用激素和抗菌药物的交叉反应小,表明该单抗具有较好的敏感性和特异性.     

抗氯霉素单克隆抗体的制备与鉴定

本文用人工合成的氯霉素 -牛血清白蛋白 (CAP- BSA)免疫 BAL B/ C小鼠 ,通过杂交瘤技术建立了 2株分泌抗氯霉素的单克隆抗体 (Mc Ab)的杂交瘤细胞 1D1 0 和 5 E6 。经间接酶联免疫吸附试验 (ci EL ISA)检测细胞培养上清效价为 1∶ 5 12 ,诱生腹水的效价可达 1× 10 8。两株单克隆抗体的亚型为 Ig G1 ,杂交瘤染色体数目为 84～ 96条。该细胞株体外传代和冻存复苏后抗体分泌稳定。ci EL ISA检测显示其与常见抗生素及结构类似物的交叉反应小 ,其灵敏度为 0 .1ng/ ml,IC50 为 2 .38ng/ m l,这表明该单抗具有较大的应用价值     

禽脑脊髓炎病毒单克隆抗体的制备及鉴定

选用AEV(VR株 )CEF适应毒制备抗原 ,免疫Balb/c小鼠。应用杂交瘤技术以间接ELISA方法筛选获得一株分泌抗AEV(VR)单克隆抗体的杂交瘤细胞。制备细胞上清及腹水进行单抗特性鉴定 ,经染色体计数、免疫球蛋白类及亚类测定、特异性试验、稳定性试验、SDS PAGE电泳 ,表明该杂交瘤细胞染色体数目介于 90～ 110之间 ,该单克隆抗体亚类为IgG2a,分子量为 142KD ,ELISA测定细胞上清效价为 1∶1× 10 4 ,腹水效价为 1∶1× 10 6,与其它病原无交叉反应 ,抗体分泌稳定。     

磺胺氯吡嗪钠单克隆抗体的制备及鉴定

应用重氮化方法制备了磺胺氯吡嗪钠(sulfachloropyrazine sodium,SPZ)完全抗原SPZ-OVA,将成功偶联的人工全抗原(SPZ-OVA)免疫BALB/c小鼠,免疫4次后,经间接ELISA检测,1号小鼠血清抗体效价达到1:6.4×104。取该小鼠脾细胞与SP2/0细胞融合。利用ELISA方法对融合后的杂交瘤细胞上清进行检测,筛选出能够稳定分泌抗体的细胞株:D9。将细胞株D9扩大培养后,经腹腔注射小鼠使其产生腹水。腹水单抗经纯化后,通过间接竞争ELISA对该单克隆抗体的效价、半数抑制浓度以及特异性进行检测。结果表明,所制备的单克隆抗体效价为1:4×105,半数抑制浓度为326 ng/m L,特异性良好。     

抗双氯青霉素单克隆抗体的制备与初步鉴定

采用碳化二亚胺法,将双氯青霉素(dicloxacillin)与牛血清白蛋白偶联制备免疫原,免疫BALB/c小鼠,取免疫小鼠的脾细胞与Sp2/0骨髓瘤细胞融合,间接ELISA筛选,阳性杂交瘤细胞用有限稀释法克隆,获得1株可稳定分泌McAb的杂交瘤细胞5D4-C9-C8;间接ELISA方法测定,其细胞上清抗体效价为1∶300,腹水效价为1∶3.5×105,为IgG1,与其结构类似物邻氯青霉素、苯唑青霉素的交叉反应率分别为17.6%和26.1%,与苄青霉素、羟氨苄青霉素和氨苄青霉素均无交叉反应性。体外传代培养和冻存复苏后抗体分泌稳定。为进一步研制检测双氯青霉素的ELISA试剂盒奠定了基础。     

猪流行性腹泻病毒M蛋白单克隆抗体的制备及鉴定

为制备猪流行性腹泻病毒(PEDV)抗M蛋白单克隆抗体(MAb),本研究以截短表达的His-M重组蛋白免疫BALB/c小鼠;以截短表达的GST-M重组蛋白作为包被抗原,采用常规的淋巴细胞杂交瘤技术制备杂交瘤细胞,通过间接ELISA进行筛选,得到一株稳定分泌抗M蛋白MAb.MAb亚类鉴定为IgG2b型,轻链为к链,杂交瘤细胞培养上清和诱导的小鼠腹水抗体效价分别为1:3000和1:2×105.Western blot试验表明该MAb能够识别重组及天然的PEDV M蛋白.间接免疫荧光试验表明该MAb能够与PEDV感染的Vero E6细胞产生特异性免疫荧光.     

抗水牛伊氏锥虫变异表面糖蛋白抗原单克隆抗体的研制

用纯化的伊氏锥虫变异表面糖蛋白（variant surface glucoprotein, VSG）免疫BALB/c小鼠,取免疫小鼠脾细胞与SP2/0骨髓瘤细胞进行融合,经过3次克隆和间接ELISA方法筛选,获得3D7、5B9 2株稳定分泌单克隆抗体的杂交瘤细胞株,用间接ELISA方法检测杂交瘤细胞培养液上清效价和小鼠腹水效价,其中细胞培养上清效价分别为1∶6400和1∶12800,腹水效价分别为1∶105和1∶106。单抗的亚型鉴定结果表明,3D7、5B9分泌的抗体都为IgG1亚类κ链。     

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.     

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.     

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.     

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.