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1.
为制备牛副流感病毒3型(BPIV3)核衣壳蛋白(NP)单克隆抗体(MAb),本研究利用原核表达并纯化的重组NP (rNP)免疫BALB/c小鼠,取免疫后小鼠脾细胞与骨髓瘤细胞SP2/0融合.采用以BPIV3为检测抗原的间接ELISA方法筛选阳性细胞克隆,经3次克隆纯化后获得1株稳定分泌抗NP特异性MAb的杂交瘤细胞株(5E5)并制备腹水,采用rNP及BPIV3包被的ELISA效价分别是2×106和1.28×105.间接ELISA、western blot、IFA试验表明该MAb具有良好的反应性和特异性.经抗体亚类鉴定该MAb亚类为IgGl/κ.特异性试验表明该MAb不与牛传染性鼻气管炎病毒、牛病毒性腹泻病毒反应.免疫组化试验表明该MAb可以检测BPIV3感染动物体内的病原.该MAb还可用于建立检测BPIV3病原及抗体的诊断方法,同时为研究NP的结构和功能提供了条件.  相似文献   

2.
SAMHD1是最近发现的在人髓系细胞中发挥抗艾滋病毒作用的一种天然蛋白.为制备抗人huSAMHD1蛋白的单克隆抗体(MAb),本研究利用原核表达并纯化的人huSAMHD1重组蛋白为免疫原,经腹腔免疫4周龄BALB/c小鼠,取其脾细胞与SP2/0骨髓瘤细胞进行融合,经间接ELISA法筛选和4次有限稀释法克隆纯化,获得了1株稳定分泌MAb的杂交瘤细胞株.Western blot与间接免疫荧光试验结果表明,获得的MAb与人huSAMHD1的真核表达蛋白呈阳性反应,而与猴、马SAMHD1的真核表达蛋白呈阴性反应,表明其具有很好的特异性,可以用于ELISA、免疫荧光和免疫印迹检测以及对huSAMHD1的进一步研究.  相似文献   

3.
为建立快速检测禽类黏膜组织中SIgA含量的方法,本研究原核重组表达鸡SIgA重链片段蛋白,纯化后免疫BALB/c鼠.以鸡胆汁中分离纯化的SIgA作为检测抗原,常规单克隆技术筛选出1株能够稳定分泌抗鸡SIgA单克隆抗体(MAb)的IgG2a、κ型MAb.经ELISA和western blot分析,所获得的1株MAb亲和力高、特异性强;采用生物素标记纯化MAb腹水IgG为检测二抗,以禽流感-新城疫重组二联活载体疫苗免疫SPF鸡为模型,初步建立了新城疫病毒和H5亚型禽流感病毒特异的黏膜SIgA间接ELISA检测方法.本研究为开展特异性家禽黏膜免疫机制的研究提供了重要技术手段.  相似文献   

4.
为制备禽传染性支气管炎病毒(IBV) S2蛋白的单克隆抗体(MAb)，本研究通过原核表达系统表达并纯化重组S2蛋白，将其免疫BALB/c小鼠并通过间接ELISA方法筛选到4株能够稳定分泌S2蛋白MAb的杂交瘤细胞株，分别命名为1A7、4F12、4A7、4D1。亚类鉴定结果显示4株MAb的重链均为Ig G1，轻链均为κ。采用间接ELISA法检测上清及腹水效价，结果显示MAb细胞上清效价均不低于1∶12 800，腹水效价均不低于1∶51 200。采用western blot检测制备的S2蛋白MAb与IBV感染鸡胚尿囊液后天然表达的S2蛋白的反应原性；采用间接免疫荧光试验(IFA)检测IBV感染非洲绿猴肾细胞(Vero)后天然表达的S2蛋白的反应原性。Western blot结果显示，感染IBV的鸡胚尿囊液中出现了90 ku左右的特异性条带；IFA结果显示，感染IBV的Vero细胞中出现绿色荧光。以上结果表明，制备的S2蛋白MAb能够与天然表达的S2蛋白反应，反应原性较强。利用间接ELISA检测MAb与S2蛋白的亲和力，结果显示4株MAb对S2蛋白均有良好的亲和力。利用一系列表达部分重叠的重...  相似文献   

5.
为制备抗蓝舌病病毒(BTV)15型VP2蛋白的单克隆抗体(MAb)及鉴定其抗原表位,本研究利用原核表达系统表达的重组VP2蛋白纯化后免疫BALB/c小鼠,将其脾细胞与SP2/0细胞进行融合,并以Bac-to-Bac昆虫杆状病毒表达系统表达并纯化的重组VP2蛋白为包被抗原,通过间接ELISA筛选出一株稳定分泌抗BTV15VP2蛋白的MAb(2D6)。通过western blot和间接免疫荧光试验对该MAb进行特异性鉴定。结果显示,其仅能够识别BTV15,而不能识别其他血清型BTV以及同种属的中山病毒和茨城病毒,表明该MAb具有良好的反应特异性。利用肽扫描技术对该MAb针对的B细胞表位进行鉴定,确定该MAb识别的抗原表位为15IAPAIIK RYP24。本研究为BTV15特异性抗原检测方法的建立奠定了基础。  相似文献   

6.
为建立鹿源牛病毒性腹泻-粘膜病病毒(BVDV)检测方法,本研究制备了抗鹿源BVDV特异性单克隆抗体(MAb)。用纯化的BVDV免疫BALB/c小鼠,取其脾细胞与SP2/0骨髓瘤细胞进行融合,经3次有限稀释法克隆和间接ELISA法筛选,获得2株稳定分泌MAb的杂交瘤细胞株2A3和4B12。通过间接ELISA检测,MAb效价为:上清液及腹水分别为1∶512、1∶640和1∶12800、1∶16000;MAb的亚类鉴定结果表明,2株杂交瘤细胞分泌的抗体亚类均为IgGl亚类;经ELISA测定,2A3和4B12与BVDVC24V株、HCV、BDV、PRV均不发生交叉反应,但与BVDVCCSYD株发生交叉反应;IFA试验结果显示,4B12和2A3与接种BVDVN71株病毒液的细胞反应产生较强的特异荧光;抗原识别位点分析结果表明,2A3和4B12两株MAb所识别的抗原位点相同。这两株MAb的获得为鹿BVDV的检测方法的建立奠定良好的基础。  相似文献   

7.
为制备抗猪瘟病毒(CSFV)单克隆抗体(MAb),本实验以表达CSFV E2囊膜糖蛋白的水泡口炎假病毒免疫BALB/c小鼠,取其脾细胞与骨髓瘤细胞SP2/0进行融合;利用间接ELISA方法和携带荧光素酶(Luciferase)报告基因的HIV-luc/CSFV-E1E2假病毒系统筛选分泌中和性E2MAb的杂交瘤细胞;测定MAb亚型并纯化后,间接ELISA方法测定MAb的效价;采用western blot鉴定MAb的特异性亲和力;利用HIV-luc/CSFV-E1E2假病毒进行体外中和试验,分析MAb抑制病毒感染的能力。结果表明本实验获得了1株分泌中和性MAb的杂交瘤细胞9C8,该MAb能与E2蛋白特异性结合,而且体外抑制试验中和效价大于1∶25600。  相似文献   

8.
p54蛋白是非洲猪瘟病毒(ASFV)进入宿主细胞的重要结构蛋白，参与病毒的早期感染，具有较强的免疫原性，常作为研制非洲猪瘟(ASF)疫苗和检测方法的重要靶点。为制备ASFV p54蛋白的单克隆抗体(MAb)，本研究通过构建重组质粒p ET28a-p54并经原核表达重组p54蛋白(rp54)，采用镍柱亲和层析方法对rp54纯化后经SDS-PAGE鉴定。结果显示，rp54主要在沉淀中出现17 ku的目的条带，经纯化效果较好。采用纯化的rp54免疫BALB/c小鼠，三免后采用间接ELISA方法测定小鼠血清的抗体效价。选择抗体效价最高的小鼠冲击免疫，3 d后取免疫小鼠脾细胞与SP2/0骨髓瘤细胞融合，通过间接ELISA筛选稳定分泌p54蛋白MAb的杂交瘤细胞。结果显示，三免后小鼠血清抗体效价最高达1∶256 000，表明该蛋白的免疫原性很好。间接ELISA筛选结果显示，获得了3株稳定分泌p54蛋白MAb的杂交瘤细胞株。分别采用western blot、间接免疫荧光试验(IFA)检测该3株MAb的反应原性，采用间接ELISA检测3株MAb的特异性。Western blot结果显示，3株MAb均在...  相似文献   

9.
为制备抗猪脑心肌炎病毒(EMCV)HB10株VP2蛋白的单克隆抗体(MAb)及鉴定其抗原表位,本研究利用原核表达系统表达的重组VP2蛋白(r VP2),纯化后将其免疫BALB/c小鼠,取免疫的小鼠脾细胞与SP2/0细胞融合,通过间接ELISA方法筛选出一株稳定分泌抗VP2 MAb(1D3)。通过间接免疫荧光试验和western blot对该MAb的免疫活性和特异性进行鉴定,结果表明1D3 MAb仅能够特异性与EMCV全病毒反应。此外,通过构建截短的VP2蛋白重组质粒,采用western blot的方法对VP2蛋白抗原表位进行鉴定,初步确定MAb 1D3识别的线性表位区域为89DGGVFGAALRRH100。本实验结果为进一步研究VP2蛋白的功能及建立EMCV检测方法奠定了基础。  相似文献   

10.
为制备赤羽病病毒(AKAV)N蛋白的单克隆抗体(MAb),本实验利用原核表达系统表达、纯化的重组N蛋白免疫BALB/c小鼠,三免后取小鼠脾淋巴细胞与SP2/0细胞融合,并以纯化的重组N蛋白为包被抗原,通过间接ELISA筛选出1株稳定分泌抗AKAV N蛋白的MAb杂交瘤细胞株IIG5,检测结果显示IIG5为IgG1/κ链。间接免疫荧光结果显示,IIG5与AKAV感染细胞呈阳性反应,与茨城病病毒、中山病病毒以及蓝舌病病毒1型均呈阴性反应,特异性较好。Western blot结果显示,IIG5能够识别重组的以及AKAV感染细胞中的N蛋白。本研究结果为AKAV检测方法的建立及相关研究奠定基础。  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.
Tropical Animal Health and Production -  相似文献   

15.
Our particular attention in this article was given to natural mediators for macrophages isolated from the sites of tissue injury. A number of chemotactic factors, which may satisfy many criteria making them acceptable as inflammatory leucocyte chemotactic factors, has been separated. Among them, our laboratory has isolated three macrophage (monocyte) chemotactic factors (MCF-a, -b and -c). Their purification, characterization and functional specificity are discussed.  相似文献   

16.
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   

17.
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   

18.
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   

19.
The objective of this report was to characterize 20-year changes in proportion of calcium oxalate (CaOx) calculi and struvite calculi in dogs, and associations with breed, age, and sex. In this retrospective study, results of analysis of urinary calculi from dogs were reviewed for specimens received between July 1, 1981, and December 31, 2001. Breed, sex, age, year of submission of the specimen, and mineral type(s) were analyzed statistically. CaOx or Struvite or both were contained in 18,966 of 20,884 (91%) specimens. For both sexes, a 20-year statistically significant increase was observed in the proportion of calculus specimens that contained CaOx. The increase in this proportion was greater in females (1% to 31%) than in males (18% to 82%). From 1998 to 2001, when proportions may have plateaued, the odds of specimens containing CaOx were markedly higher in 18 breeds, markedly lower in 5 breeds, and not significantly different in 13 breeds compared with crossbreds. For both sexes, a 20-year statistically significant decrease was observed in the proportion of calculus specimens that contained struvite. This decrease in proportion was greater for males (79-16%) than for females (97-68%). From 1998 to 2001, when proportions plateaued, the odds of calculi containing struvite were markedly lower in 20 breeds, markedly higher in 1 breed, and not significantly different in 15 breeds when compared with crossbreds. Breed, age, and sex were associated statistically with CaOx or struvite urolithiasis. In conclusion, there appears to have been a long-term increase in the proportion of specimens of canine urinary calculi that contain CaOx as well as a long-term decrease in the proportion of specimens of calculi that contain struvite for both male and female dogs. The rate of change appeared to begin leveling off in the period 1998 to 2001. The recent proportion of dogs with either CaOx- or struvite-associated urolithiasis may depend on breed, age, and sex, and on interactions among these 3 factors.  相似文献   

20.
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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