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1.
运用Mannich的反应,将褪黑激素(MLT)与牛血清白蛋白(BSA)连结,制备MLT完全抗原。MLT与BSA连结比为12:1。将此完全抗原免疫5只雄兔,制备特异性抗体;将MLT用^125I标记,薄层层析筛选标记物,制备高纯度的放射性标记物,建立竞争性MLT-RIA法。本法可测范围为15.31-1500pg,灵敏度为每管15.31pg,批内变异系数6.7%,批间变异系数14.8%。用此法测定夏至时考力代羊昼夜血样,测定结果与国外基本一致;测定夏至时湖羊昼夜血样,发现湖羊MLT分泌有其特殊性,即血液MLT水平呈明显的昼夜变化;白天为100ng/L左右,而黑暗开始时,MLT水平迅速上升,在20:00-次日8:00处于高水平,最大值为1100ng/L左右。  相似文献   

2.
用牛血清白蛋白(BSA)免疫普通级试验兔(新西兰白兔),获得高滴度的抗BSA血清。以系列含量的BSA溶液绘制标准曲线,测定兔血清中抗BSA抗体效价。结果表明:普通级试验兔BSA4次免疫后,兔血清中抗BSA抗体效价为1:9000。ELISA间接法敏感度高,优化了酶免疫反应条件,标准曲线线性范围内r=0.9965,可初步用于抗BSA抗体含量检测。  相似文献   

3.
犬瘟热病毒抗体检测方法的比较研究   总被引:19,自引:0,他引:19  
用犬瘟热(CD)弱毒和自制的高免犬血清,建立了检测犬瘟热病毒(CDV)抗体的中和(SN)试验、间接荧光抗体技术(IFAT)和间接免疫酶染色法。这3种方法对38份CD弱毒疫苗免疫犬血清抗体效价的测定结果表明,当被检血清稀释度为1∶100时,SN试验、IFAT和间接免疫酶染色法测定时的阳性血清数分别为29,7和31份,IFAT和间接免疫酶染色法测定结果相对于SN试验结果的符合率分别为42.1%和94.7%。同时发现,当SN试验测定的血清效价在1∶100以上时,IFAT测定结果总是在1∶25以上,间接免疫酶染色法测定结果总是在1∶100以上。3种方法以各自初步确定的CDV血清抗体效价完全保护值指标计算所测血清的完全保护率,SN试验为76.3%,IFAT为78.9%,间接免疫酶染色法为81.6%。据此认为,IFAT和间接免疫酶染色法均可部分代替SN试验用于CD疫苗免疫效果的评价、免疫犬抗体水平的监测以及CD的流行病学调查  相似文献   

4.
猪金属硫蛋白抗体的制备及应用   总被引:4,自引:0,他引:4  
以自猪肝提取的Zn—MT与BSA的偶联物为抗原,免疫昆明小鼠制备了多克隆抗体。获得的抗体经鉴定后用于建立检测猪MT的间接竞争EI。ISA。结果表明:制备的IgG达到电泳纯,是猪MT的特异性抗体。所建立的间接竞争ELISA的灵敏度为4.1ng/mL,批内变异系数是9.976%,批间变异系数为14.3858%,可用于猪体MT的定性、定量测定。  相似文献   

5.
将多西环素(doxycycline,DOX)进行化学修饰引入羧基或氨基等活性基团,然后与牛血清白蛋白(BSA)和卵清蛋白(OVA)偶联合成人工免疫原DOX—PABA—BSA、DOX—BSA和包被原DOX—PABA—OVA、DOX~OVA,并用紫外吸收(UV)、凝胶电泳(SDS—PAGE)和EUSA方法对人工抗原进行鉴定;将合成的人工抗原DOX—PABA—BSA、DOX—BSA分别免疫BALB/C小鼠,用间接ELISA方法测定多抗(pAb)效价,用竞争ELISA方法鉴定其敏感性,用交叉反应试验鉴定其特异性。结果表明,二个免疫组6只小鼠血清抗体效价均在1:6400以上,DOXpAb对DOX的50%抑制浓度(IC50)在39.79~53.13μg/L,抗血清与四环素类药物交叉反应很低。本实验为建立多西环素ELISA残留免疫学检测方法和并制备多西环素试剂盒奠定了基础。  相似文献   

6.
氯霉素全抗原合成及多克隆抗体的制备   总被引:4,自引:0,他引:4  
用重氮化方法将氯霉素与牛血清白蛋白(BSA)和卵清蛋白(OVA)偶联,制备免疫抗原和包被抗原,用ELISA方法进行鉴定。将偶联抗原用弗氏佐剂乳化后免疫德国大白兔。制备多克隆抗体,并用硫酸铵沉淀法初步纯化。用亲和层析法进一步纯化。用所得抗体建立竞争酶联免疫吸附分析法(ELISA)用于氯霉素检测,最低检查限为0.3ng/mL。  相似文献   

7.
应用商品化酶联免疫吸附试验(ELISA)试剂盒和血球凝集抑制(HI)试验同时测定187份常规鸡新城疫(ND)抗体血样。结果显示,二种方法的抗体效价呈显著相关,其相关系数为0.83。在此基础上,以HI法为标准方法对ELISA的临床检测结果进行了分析。HI法的测定结果表明,该试验群的抗体阳性率为54.5%。按试剂盒提供的阴、阳性临界点判定结果,ELISA的相对灵敏度为97.1%,相对特异性为61.2%  相似文献   

8.
酶联免疫法测定鸡肉中尼卡巴嗪残留   总被引:1,自引:0,他引:1  
为了建立鸡肉中尼卡巴嗪残留标志物4,4’-二硝基均二苯脲(DNC)残留酶联免疫(ELISA)检测方法。将N-琥珀酰-L-丙氨酰-L-丙氨酰-L-丙氨酸4-硝基苯胺(SAN)与载体蛋白偶联作为抗原免疫动物,获得抗DNC抗体,在此基础上建立了鸡肉中尼卡巴嗪残留的检测方法。人工抗原中SAN与载体蛋白的结合比约为12:1,血清效价1:2000倍,鸡肉中检测限为9.2μg/kg,添加回收率在49.4%~118%之间,批内、批间变异系数均小于20%。该方法在灵敏度、精密度和准确度方面均能满足兽药残留筛选检测要求。  相似文献   

9.
抽取7批牛口蹄疫Asia—Ⅰ型灭活疫苗,用OIE推荐的50%保护(PD50)试验方法进行免疫保护试验;同时,用OIE推荐的液相阻断酶联免疫吸附试验(LPB—E)检验程序测定了7批疫苗免疫保护试验的103头牛的血清抗体滴度。经过对获得数据的统计和处理分析证明,LPB—E抗体滴度与牛免疫保护率存在着显著的正相关性(P〈0.05),相关系数为0.91,概率分析表明,LPB—E抗体滴度≥2.1(1:126)时,95%的免疫牛能够抵抗与疫苗毒株同源的10^4ID50强毒的攻击。  相似文献   

10.
福州市郊某猪场于1995年11月~1996年4月期间哺乳仔猪大多发病,此间共死亡3000余头,造成直接经济损失数十万元,后经诊断为猪瘟。为配合该猪场开展猪瘟强毒的净化工作,我们采集了148份血样,应用猪瘟单抗诊断试剂进行猪瘟强毒抗体的检测,结果阳性者33份,阳性检出率22.3%。现将其试验报道如下:互.材料与方法1.1材料1.1.五猪瘟单折诊断试剂与酶联板:购自中国兽药监察所。1.l.2常规酶联反应试剂:包括包被液、磷酸缓冲液(PBS)、PBS一吐温洗涤液(PBS-T)、稀释液、邻苯二胺底物溶液(OPD)和终止液,自配。1.1.3待检…  相似文献   

11.
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。  相似文献   

12.
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.  相似文献   

13.
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.  相似文献   

14.
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.  相似文献   

15.
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.  相似文献   

16.
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.  相似文献   

17.
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.  相似文献   

18.
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.  相似文献   

19.
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.  相似文献   

20.
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