洛美沙星人工抗原的合成及鉴定

通过化学方法制备了洛美沙星的免疫抗原和包被抗原;分别采用碳二亚胺（EDC）法和氯甲酸乙丁酯法把洛美沙星与BSA进行偶联制备了人工免疫抗原LOM-BSA,EDC法制备了包被抗原LOM-OVA,经非变性PAGE、紫外分光光度法测定了人工合成抗原的偶联比,EDC法和氯甲酸乙丁酯法制得的LOM-BSA两分子结合比率分别为4∶1和11∶1,LOM-OVA中两分子结合比率为12∶1。这为洛美沙星单克隆抗体的制备奠定了基础。     

克伦特罗小分子半抗原偶联率的测定方法研究

利用重氮化两步法将半抗原盐酸(CL)分别偶联到牛血清蛋白(BSA)和卵清蛋白(O-VA)上制得了免疫抗原BSA-CL和包被抗原OVA-CL。建立了用紫外分光光度法测定偶联比率的方法,测得它们的偶联率分别为39:1和24:1。     

酸性红73人工抗原的合成与鉴定

为进一步制备酸性红73新型人工抗原,试验采用N,N-羰基二咪唑(CDI)法合成酸性红73的免疫原和包被原,经紫外扫描和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)鉴定偶联成功,酸性红73与牛血清白蛋白(BSA)、卵清蛋白(OA)的偶联比分别为9∶1和5∶1。免疫后所得兔源多克隆抗体,采取双向琼脂扩散实验和方阵滴定法确定包被抗原和抗体的最适工作浓度分别为1∶4000、1∶8000。     

两种不同偶联比的莱克多巴胺免疫原免疫效果比较

将同窝3月龄体重2 kg左右的新西兰白兔随机分为2组,每组5只,分别用偶联比为2.5∶1、10∶1的莱克多巴胺-BSA(BSA-Rac)免疫6次后,间接ELISA和间接抑制ELISA检测抗血清的效价和特异性。结果表明,偶联比为10∶1的免疫原产生了效价超过1∶10000以上、IC₅₀值约为10 ng/ml的抗体,而偶联比为2.5∶1的免疫原未产生抗体。     

多西环素人工抗原的合成与鉴定

建立多西环素人工抗原(Doxycycline,DOX)的合成及鉴定方法。采用戊二醛偶联法将DOX分别与载体牛血清白蛋白(BSA)及卵清蛋白(OVA)偶联制备免疫抗原(DOX-BSA)和包被抗原(DOX-OVA)。经SDS-PAGE电泳法、紫外可见吸收光谱鉴定,证实人工抗原成功合成,偶联比分别为8.6∶1和3.8∶1。结果表明建立了一种新的DOX抗原合成方法,为进一步制备特异性DOX抗体和建立检测食品中DOX的酶联免疫方法奠定了基础。     

赭曲霉毒素A完全抗原的制备及鉴定

赭曲霉毒素A(OTA)是一种对人和动物具有广泛毒性的霉菌毒素,为建立OTA在食品和饲料中的快速检测方法,采用活性酯法(NHS)将OTA与牛血清白蛋白(BSA)、鸡卵清白蛋白(OVA)偶联,分别制备了免疫原OTA-BSA和包被原OTA-OVA。紫外扫描和SDS-PAGE鉴定表明,OTA与BSA和OTA与OVA偶联成功,偶联物OTA-BSA和OTA-OVA的偶联比分别为7∶1和4.5∶1。用免疫原OTABSA免疫Balb/c小鼠,制备多克隆抗体,用OTA-OVA包被ELISA板进行检测,小鼠血清中抗OTA的抗体效价最高可达1∶51 200。表明制备的免疫原OTA-BSA具有良好的免疫原性,为OTA单克隆抗体的制备奠定了基础。     

肉兔体内猪瘟抗体滴度对猪瘟定型热比率的影响

以猪瘟正向间接血凝法对肉兔进行猪瘟抗体水平的检测,选用猪瘟抗体滴度≥1∶4( )的试验兔9群686只、0~1∶2( )试验兔2群328只。每只兔注射含4000个发热单位的猪瘟兔化弱毒(脾毒乳浆液)。结果表明猪瘟抗体滴度≥1∶4( )的试验兔定型热比率加权平均值为63.3%、0~1∶2( )试验兔定型热比率加权平均值96.3%,差异极显著。以4个兔场12511只肉兔作扩大试验,结果表明无猪瘟抗体试验兔(占94.8%)的平均定型热比率为88.6%;猪瘟抗体滴度≥1∶4( )试验兔(占56.7%)定型热比率为79.1%。     

用于检测孔雀石绿的全抗原的合成与鉴定

采用重氮化法,将半抗原副品红(Pararosaniline,PA)分别与载体蛋白牛血清白蛋白(BSA)、人血清白蛋白(HSA)和卵清蛋白(OVA)偶联,反应得到免疫抗原和包被抗原,其偶联最佳结合比分别为8.8∶1,12.7∶1,6.9∶1,经紫外扫描光谱和蛋白电泳鉴定得到全抗原。用制备的抗原乳化后免疫小鼠获得了效价较高的多克隆抗体。为进一步制备抗孔雀石绿单克隆抗体打下基础。     

苯乙醇胺A人工抗原的合成及鉴定

采用重氮化法将苯乙醇胺A分别与牛血清白蛋白(BSA)和卵清白蛋白(OVA)偶联,制备免疫抗原苯乙醇胺A(PEAA)-BSA和检测抗原PEAA-OVA。经紫外扫描法和SDS-PAGE鉴定表明偶联成功,偶联比分别为15∶1、17.4∶1。用PEAA-BSA免疫BALB/c小鼠,采用间接ELISA法检测抗体效价,采用竞争ELISA法检测IC50值,获得了效价4.1×105以上、IC50值为0.2μg/L的多抗血清,证明合成了具有免疫原性的人工免疫原。     

赭曲霉毒素A人工抗原合成及多克隆抗体的制备

试验采用活性酯法将赭曲霉毒素A(OTA)分别与BSA、KLH偶联,合成人工免疫抗原OTA-BSA和人工检测抗原OTA-KLH,并通过紫外光谱扫描(UV)、SDS-PAGE验证偶联成功,OTA-BSA的偶联比为12.5∶1,OTA-KLH的偶联比为10∶1。用OTA-BSA免疫BALB/c小鼠,获得高效价、高特异的小鼠多抗血清,多抗血清效价达1∶2×105以上,在0.1 ng/m L~8.1 ng/m L的范围内,线性关系良好,R2=0.996 1,IC50为0.18 ng/m L。鼠多抗血清除与赭曲霉毒素B有18%的交叉反应率外,与玉米赤霉烯酮、黄曲霉毒素B 1、脱氧雪腐镰刀菌烯醇等类似物的交叉反应率(CR)均小于0.2%,特异性良好。     

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.     

Phase I evaluation of CCNU (lomustine) in tumor-bearing cats

1-(2-Chloroethyl)3-cyclohexyl-1-nitrosourea (CCNU) is an alkylating agent in the nitrosourea subclass. A prospective evaluation of CCNU was done to determine the maximally tolerated dosage of CCNU in tumor-bearing cats. Response data were obtained when available. Twenty-five cats were treated with CCNU at a dosage of 50-60 mg/m3 body surface area. Complete hematologic data were available for 13 cats. Neutropenia was the acute dose-limiting toxicity. The median neutrophil count at the nadir was 1,000 cells/microL (mean, 2,433 cells/microL; range, 0-9,694 cells/microL). The time of neutrophil nadir was variable, occurring 7-28 days after treatment, and counts sometimes did not return to normal for up to 14 days after the nadir. Based on these findings, a 6-week dosing interval and weekly hematologic monitoring after the 1st treatment with CCNU are recommended. The nadir of the platelet count may occur 14-21 days after treatment. The median platelet count at the nadir was 43,500 cells/microL. No gastrointestinal, renal, or hepatic toxicities were observed after a single CCNU treatment, and additional studies to evaluate the potential for cumulative toxicity should be performed. Five cats with lymphoma and 1 cat with mast cell tumor had measurable responses to CCNU. Phase II studies to evaluate antitumor activity should be completed with a dosing regimen of 50-60 mg/m3 every 6 weeks.     

Prevalence of gastric lesions in racing Alaskan sled dogs

Human and equine athletes are reported to have a high prevalence of gastric disease, and anecdotal evidence suggests a similar phenomenon applies to racing sled dogs. To investigate the prevalence of gastric disease in racing sled dogs, we conducted 2 gastroscopy studies on dogs competing in the annual Iditarod Sled Dog Race. A pilot study of dogs that were either dropped from the 2000 Iditarod Sled Dog Race because of illness or that finished the race indicated that, approximately 5 days after competing, 10 of 28 dogs (35%) had endoscopic evidence of gastric ulceration, erosion, or hemorrhage. The next year, an endoscopic study of 73 dogs participating in the 2001 Iditarod race was performed in order to evaluate a larger population of dogs. Data from 70 of these dogs could be used; 34 (48.5%) had ulceration, erosion, gastric hemorrhage, or some combination of these findings. When this group of 70 dogs was compared retrospectively to a control group of 87 dogs presented to the Texas A&M University (TAMU) Veterinary Medical Teaching Hospital, the Iditarod sled dogs had a significantly higher prevalence (P = .049) of gastric lesions. These findings suggest that, similar to athletes of other species, elite canine athletes have an increased prevalence of gastric disease compared to the canine population at large.     

Furosemide continuous rate infusion in the horse: evaluation of enhanced efficacy and reduced side effects

Continuous rate infusion (CRI) of furosemide in humans is considered superior to intermittent administration (IA). This study examined whether furosemide CRI, compared with IA, would increase diuretic efficacy with decreased fluid and electrolyte fluctuations and activation of the renin-angiotensin-aldosterone system (RAAS) in the horse. Five mares were used in a crossover-design study. During a 24-hour period, each horse received a total of 3 mg/kg furosemide by either CRI (0.12 mg/kg/h preceded by a loading dose of 0.12 mg/kg IV) or IA (1 mg/kg IV q8h). There was not a statistically significant difference in urine volume over 24 hours between methods; however, urine volume was significantly greater after CRI compared with IA during the first 8 hours ([median 25th percentile, 75th percentile]: 9.6 L [8.9, 14.4] for CRI versus 5.9 L [5.3, 6.0] for IA). CRI produced a more uniform urine flow, decreased fluctuations in plasma volume, and suppressed renal concentrating ability throughout the infusion period. Potassium, Ca, and Cl excretion was greater during CRI than IA (1,133 mmol [1.110, 1,229] versus 764 mmol [709, 904], 102.7 mmol [96.0, 117.2] versus 73.3 mmol [65.0, 73.5], and 1,776 mmol [1,657, 2.378] versus 1,596 mmol [1,457, 1,767], respectively). Elimination half-lives of furosemide were 1.35 and 0.47 hours for CRI and IA, respectively. The area under the excretion rate curve was 1,285.7 and 184.2 mL x mg/mL for CRI and IA, respectively. Furosemide CRI (0.12 mg/kg/h) for 8 hours, preceded by a loading dose (0.12 mg/kg), is recommended when profound diuresis is needed acutely in horses.     

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.     

The Epidemiology of Gastrointestinal Nematodes of Dairy Cattle in Central Kenya

The epidemiology of H. placei and of other gastrointestinal nematodes in yearling dairy cattle was examined on two farms in Kiambu District, central Kenya during each of 13 one-month periods from April 1993 to April 1994. On each farm, 32 newly weaned dairy calves were given a single dose of albendazole and then placed on experimental pastures. Twelve of the animals were designated for bi-monthly slaughter (n = 2) and analysis of worm population characteristics and 20 were designated for blood and faecal collection and for weighing. Two parasite-free tracer calves were grazed alongside the weaner calves each month throughout the study period and were also slaughtered for analysis of worm populations. Faecal egg counts, haematological and serum pepsinogen determinations, herbage larval counts, and animal live weight changes were recorded monthly. The study revealed that Haemonchus placei, Trichostrongylus axei, Cooperia spp. and Oesophagostomum radiatum were responsible for parasitic gastroenteritis and that H. placei was the predominant nematode present in the young cattle on both farms. Faecal egg counts from resident cattle and necropsy worm counts revealed that pasture larval levels were directly related to the amount of rainfall. The total worm burdens in the animals were highest during the rainy season (March–June and October–December) and lowest during the dry seasons (July–September and January–February). The very low recovery of immature larvae of H. placei from the tracer calves indicated that arrested development is not a feature of the life cycle of this parasite in central Kenya. The maintenance of the parasite population depended on continuous cycling of infection between the host and the pasture. The agroclimatic conditions of the study area were such that, in general, favourable weather conditions for the development and survival of the free-living stages of gastrointestinal nematodes existed all year round.     

Diversity of Ectoparasites in Sheep Flocks in Sa~o Paulo,Brazil

The occurrence of ectoparasites in sheep flocks is frequently reported but seldom quantified. Sheep production used to be a predominantly family activity in the state of Sa~o Paulo (Brazil), but it began to become a commercial activity in the past decade. Thus, information about the ectoparasites existing in sheep flocks has become necessary. The present data were obtained by means of questionnaires sent to all sheep breeders belonging to the `Associaça~o Paulista de Criadores de Ovinos' (ASPACO; Sa~o Paulo State Association of Sheep Breeders). Response reliability was tested by means of random visits paid to 10.6% of the respondents. Most of the properties (89.5%) reported the presence of one or more ectoparasites. Screw-worm (Cochliomyia hominivorax) was the most frequent ectoparasite (72.5%), followed by bot fly larvae (Dermatobia hominis, 45.0%), ticks (Amblyomma cajennense) and Boophilus microplus, 31.3%) and finally lice (Damalinia ovis, 13.8%). Combined infestations also occurred, the most common one being screw-worm with bot fly larvae (36.0%) followed by bot fly larvae with ticks (13.9%), screw-worm with ticks (9.3%), bot fly larvae with lice (6.9%), and ticks with lice (5.0%). The most common triple combination was screw-worm, bot fly larvae and ticks (12.8%). Breeds raised for meat or wool were attacked by bot fly larvae and ticks more often than other breeds. Lice were only absent from animals of indigenous breeds. The relationships among these ectoparasites are discussed in terms of sheep breeds, flock size, seasonality and the ectoparasitic combinations on the host.     

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.