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为研究日本血吸虫丝氨酸-苏氨酸蛋白磷酸酶（SjPP）的免疫功能，构建了原核表达重组质粒pET28a（＋）-SjPP，转化大肠埃希氏菌BL21（DE3）进行诱导表达，并用重组蛋白进行了小鼠免疫保护试验，免疫剂量为每次20μg／只，共免疫3次。结果表明，pET28a（＋）-siPP／BL21（DE3）在0．1mmol／L IPTG诱导2h时，每1g菌体可产生17．8mg以包涵体形式存在的重组蛋白；重组蛋白免疫小鼠后诱导产生了高效价的血清特异性IgG抗体，并获得了部分减虫率、肝组织减卵率和显著的粪便减卵率，具有一定的免疫保护作用。 相似文献
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利用PCR技术扩增编码水疱性口炎病毒印第安纳型（VSV—Ind）糖蛋白基因的主要抗原表位区，通过引入其两端的EcoRⅠ和XhoⅠ酶切位点，定向插入到pET30c质粒T7启动子下游的多克隆区，构建重组质粒pET30c—vsvG，并转化至宿主菌BL21（DE3）株中。用1mmol／LIPTG诱导后，SDS—PAGE电泳表明，重组蛋白在大肠埃希氏菌中得到了高效表达，经过4h表达即可达到高峰；融合蛋白的相对分子质量为57ku。重组蛋白含有六聚组氨酸尾，主要以包涵体形式存在，Ni柱纯化后的蛋白经Western—blotting鉴定，具有良好的抗原性和特异性。 相似文献
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利用PCR方法扩增乳源致病性金黄色葡萄球菌nEBPS全基因，将其定向克隆至原核表达载体pET30a（＋）中，鉴定正确后，转化BL21表达菌，经IPTG诱导获得了以可溶性表达的重组蛋白。通过NiNTAPurificationSystem纯化重组蛋白，纯化蛋白质量浓度为2．16g／L。纯化蛋白经免疫印迹检测显示重组蛋白能够被牛源金黄色葡萄球菌阳性血清识别，具有良好的反应性。将纯化的重组蛋白免疫新西兰白兔制备多克隆抗体，间接EI，1SA测定抗体效价为1：25600，凝集试验测定抗体效价为1：128。 相似文献
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将V13KL编码基因片段克隆到原核表达载体pET30b（＋），并在目的蛋白N端设计肠激酶酶切位点，构建出pET30b（＋）-VK13L重组质粒，转化大肠杆菌表达菌株BL21（DE3），优化诱导条件，使得抗菌肽得到了高效表达。经Tricine-SDS-PAGE和Westernblotting鉴定，目的蛋白的相对分子质量与预期一致。结果表明，V13KL基因成功克隆并且获得表达，在诱导体系中加入1mmol／LIPTG诱导6h便可检测到蛋白表达。 相似文献
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为在大肠杆菌表达系统中高效表达牛α-干扰素(bovine interferon-α,Bo IFN-α),将经密码子优化后的牛α-干扰素成熟蛋白基因合成并克隆到表达载体p Cold II中,转化大肠杆菌BL21(DE3)中诱导表达,获得可溶性的表达蛋白。对重组菌表达条件进行优化,结果显示在IPTG浓度为0.64 mmol/L,诱导温度为15℃,诱导时间为9 h的条件下,可溶性目的蛋白的表达量最高。重组蛋白经镍柱纯化后的纯度为99.2%,表达量为36 mg/L菌液。重组蛋白在MDBK细胞上抗水泡性口炎病毒的活性为1×106U/mg。 相似文献
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鸡β-防御素-1基因(Gal-1)在大肠杆菌中的融合表达与纯化 总被引：2，自引：0，他引：2
内源性抗菌多肽防御素是机体先天性免疫系统的重要组成部分，可帮助机体防御外界病原微生物的入侵。鸡β防御素-1（Gallinaein-1，Gal-1）对许多致病菌都具有杀菌作用，具有很好的研究开发价值。本研究利用PCR技术从重组质粒pGEM-TEasy-gal-1中克隆出Gal-1基因成熟肽区，将该基因插入原核表达载体pET-32a（＋）中，构建重组质粒pET-32a（＋）-gal-1，然后用重组质粒转化大肠杆菌BL-21。挑选阳性克隆菌，用浓度为0．5mmol／L的IPTG进行诱导表达。SDS-PAGE电泳显示在目标位置约25kD处出现了条带并用Werstern Blot进行了鉴定。表明Gal-1基因在大肠杆菌以融合形式得到了表达。经灰度扫描显示重组蛋白表达量占细菌总蛋白的15．54％。用50％Ni-NTA纯化树脂进行层析纯化，在洗脱液E中出现单一条带。Gal-1多肽在pET-32a（＋）质粒表达体系中成功表达为鸡防御素多肽下一步的研究奠定基础。 相似文献
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利用PCR方法扩增产单核细胞李斯特菌（LM）的溶血素基因hlyA,获得一条大小约为1600bp的目的片段,将其与pMD18-T载体连接,经酶切、PCR鉴定,命名为pMD18-T—hlyA。测序结果显示所扩增的hlyA基因与GenBank中发表的hlyA的序列同源性为99％。利用含有BamHⅠ／XhoⅠ酶切位点引物B从pMD18-T—hlyA扩增不含信号肽序列的目的片段B,获得了大小约为1500bp的基因片段,该片段与pET32a表达载体经BamHⅠ／XhoⅠ处理后进行连接,转化BL21受菌体。通过酶切、PCR鉴定及序列测定后获得阳性pET32a-hlyA表达重组质粒,将重组菌用终浓度为0．1mmol／L的IPTG进行诱导表达,结果表明重组菌在诱导2h～3h表达量最高,融合蛋白约占菌体的40％,分子量大小约为80ku;利用LM的阳性血清进行Western blot分析,证实该融合蛋白可以被LM阳性血清所识别,为今后研制针对该蛋白的单克隆抗体和建立间接ELISA诊断方法提供了条件。 相似文献
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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
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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
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Rassnick KM Gieger TL Williams LE Ruslander DM Northrup NC Kristal O Myers NC Moore AS 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2001,15(3):196-199
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
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Davis MS Willard MD Nelson SL Mandsager RE McKiernan BS Mansell JK Lehenbauer TW 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2003,17(3):311-314
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
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Furosemide continuous rate infusion in the horse: evaluation of enhanced efficacy and reduced side effects 总被引：1，自引：0，他引：1
Johansson AM Gardner SY Levine JF Papich MG LaFevers DH Fuquay LR Reagan VH Atkins CE 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2003,17(6):887-895
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
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B. Cinar Kul N. Bilgen J.A. Lenstra O. Korkmaz Agaoglu B. Akyuz O. Ertugrul 《Zeitschrift für Tierzüchtung und Züchtungsbiologie》2015,132(6):449-453
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
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Waruiru RM Thamsborg SM Nansen P Kyvsgaard NC Bogh HO Munyua WK Gathuma JM 《Tropical animal health and production》2001,33(3):173-187
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
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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
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Troxel MT Vite CH Van Winkle TJ Newton AL Tiches D Dayrell-Hart B Kapatkin AS Shofer FS Steinberg SA 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2003,17(6):850-859
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