鹅枕静脉窦长期大量采血

将１２０只青年鹅分为４组，每组３０只。Ⅰ组为对照组，不进行免疫和采血，Ⅱ、Ⅲ、Ⅳ组进行３个采血期的免疫和鹅枕静脉窦采血，每个采血期采血３次，每２ｄ１次，５０ｍＬ／只。结果表明，在不同饲养条件下，Ⅱ、Ⅲ、Ⅳ组９０只鹅３个采血期的平均采血量在４１０ｍＬ／只以上，且血清抗体效价稳定；在３个采血间期内，Ⅱ、Ⅲ、Ⅳ组鹅的体重恢复率分别为９８．４％、９９．８５％和１００．７５％。由此认为，在同一群鹅内，可进行数个采血期的免疫和枕静脉窦大量采血，且不会影响血清的抗体效价。     

鹅大量采血体重变化的测定

鹅大量采血体重变化的测定周岩许万祥李立顺汪夏时（安徽农业技术师范学院凤阳２３３１００）制作禽类高免血清最大的经济成本和难题是大量屠宰放血后禽尸的处理。我们曾介绍了鹅枕静脉窦采血制抗小鹅瘟血清的方法〔１〕。但对采血中及采血后鹅群体重变化在国内外尚少见报...     

鹅抗ND,IBD双联高免血清的研制与应用

用ＩＢＤ和ＮＤ油乳剂免疫原，经３次免疫鹅后，可获得ＩＢＤ琼扩抗效价为１：６４，ＮＤ血抑制抗体效价为１：２０４８的双联高免鹅血清。     

羔羊腺病毒感染的血清学调查

连续两年在衣阿华州及其附近各州进行轿清学调查，以了解绵羊感染腺病毒１－４型和牛腺病毒２、３和７型的感染率和血清转化率，对１－３月龄羔羊采血分离血清，２月后再次采血样，应用微量血清中和试验检测抗休。第一次采集的血清样品，除ＢＡＶ－３外，其他型抗体阳性率（效价≥２）都很高，一些羔羊的抗体价增加了４倍，感染率和血清转化率分别为，ＯＡＶ－１：９４和７．２％；ＯＡＶ－２：９８．２和１５．１：ＯＡＶ－３：８５     

鹅抗ND、IBD双联高免血清的研制与应用

用ＩＢＤ和ＮＤ油乳剂免疫原，经３次免疫鹅后，可获得ＩＢＤ琼扩（ＡＧＰ）抗体效价为１：６４，ＮＤ血抑制（ＨＩ）抗体效价为１：２０４８的双联高免鹅血清．鹅在第３次免疫后１５天，血清中抗体效价开始达到高峰．并至少可维持１０天以上。经临床应用表明，抗ＩＢＤ－ＮＤ双联高免鹅血清对ＩＢＤ单纯性感染病鸡群的治愈率平均为９５．１％，对ＩＢＤ、ＮＤ混合感染病鸡群的治愈率平均为８８．２％。     

抗小鹅瘟病毒中和性单克隆抗体研制及实验防治效果

用淋巴细胞杂交瘤技术研制出１１株抗小鹅瘟病毒（ＧＰＶ）单克隆抗这些单抗仅与ＧＰＶ反应，与其它细小病毒（ＰＣＰＶ、ＦＰＶ、ＰＰＶ）和其它禽病毒（ＮＤＶ，ＩＢＤＶ，ＤＨＶ、ＤＰＶ）均不反应，腹水单抗的ＥＬＩＳＡ效价达１０＾－５－１０＾－７，琼扩沉淀效效价达１：２０－１：５１２，鹅胚中和效价达１：３０－１：１２２，鹅体中和效价为１：５４－－１：９６。ＧＰＳ１和ＧＰＧ５２株单抗对人工感染ＧＰＶ的雏鹅具有良     

犬冠状病毒超免疫血清的制备及应用

采用犬冠状病毒（ＣＣＶ）弱毒株和强毒株多次免疫健康犬，采集超免疫血清，其ＣＣＶ血清中和（ＳＮ）抗体效价高达１：２１０。试验结果表明，该超免疫血清安全、特异，无毒副作用，每只幼犬注射１．５ｍＬ／ｋｇ体重的该血清即可使９０％以上的犬获得１０￣１５ｄ的被动免疫保护；对１１０只患ＣＣＶ性肠炎病犬的临床治疗结果表明，该超免疫血清治疗效果可靠，其治愈率达９４％。     

水牛抗小鹅瘟高兔血清的研制和应用

用小鹅病毒浓缩乳化抗原，皮内多点注射分３次免疫水牛，可获得琼扩抗体为１：６－１：６４的抗小鹅瘟高免血清，此血清具有良好的预防效果和较高疗效，在疫区注射１－５日龄雏鹅预防小鹅瘟，可提高雏鹅成活率３６．４％；治疗发病雏鹅，治愈率达８８．７％。乳化抗原以首免后血清琼扩抗体效价高低而定，抗体高加倍剂量大，低则加倍剂量小。同时测得水牛三免后１３－１６天血清琼扩抗体开始达高峰。血清达高峰后，至少可以持续１６天     

鹅细小病毒免疫血清的制备及其应用

应用鹅细小病毒(GPV)弱毒疫苗及灭活疫苗免疫111只2月龄鹅群，制备了GPV免疫血清，第二次免疫后68 d采取血清，经检测ELISA抗体效价达1：8000以上，经保护试验后初步应用于鹅细小病毒感染的田间防治，收到了良好的疗效。     

用间接血凝试验检测猪传染性胸膜肺炎

经戊二醛化－鞣酸化处理的绵羊红细胞用胸膜肺炎嗜血杆菌（ＨＰ）抗原致敏,以间接血凝试验（ＩＨＡ）检测猪传染性胸膜肺炎病血清抗体。致敏红细胞抗原的最佳浓度为１００－１５０μｇ／ｍＬ,超免疫血清的抗体效价达１：５１２－１：１０２４,对人工感染１４ｄ的猪血清阳性检出率达８０％,与猪瘟、猪肺疫、喘气病、猪萎缩性鼻炎阳性血清无效叉反应。     

兔病毒性出血症病毒高免血清的制备与应用效果

以兔病毒性出血症病毒组织灭活疫苗免疫家兔，经五次免疫之后，抗体效价可达12log2以上，采集兔血清，以此治疗攻毒兔，结果大部分兔都存活，临床应用效果表明：以此血清治疗自然感染兔病毒性出血症病毒的发病兔，治愈率达80％以上。     

Partial passive protection with two monoclonal antibodies and frequency of feeding of hyperimmune anti-transmissible gastroenteritis virus (TGEV) serum for protection of three-day-old piglets from a TGEV challenge infection.

Passive protection experiments were conducted to determine the frequency and amounts of hyperimmune antiserum needed to block a transmissible gastroenteritis virus (TGEV) challenge infection and to identify monoclonal antibodies that are partially protective against TGEV. Hyperimmune antiserum or monoclonal antibodies were added to milk at each feeding or at selected feedings when the amount of antiserum was reduced. Three-day-old piglets were challenged with virulent virus that had been preincubated with antiserum or monoclonal antibodies. The results indicated that supplementing antiserum every other day was not efficacious for protection. Supplementing even small quantities of hyperimmune antiserum (0.5 ml) at least once a day in most cases was sufficient for piglet survival but did not prevent morbidity. Increasing the amount (>2 ml) and providing antiserum 3 times/day completely blocked the TGEV challenge infection. Two monoclonal antibodies were discovered that also provided passive protection for baby pigs. One monoclonal antibody, 5G1, had a high neutralizing titer, and the other, 6C4, was more effective in neutralizing and binding to virulent TGEV than to attenuated TGEVs. Both of these monoclonal antibodies were partially effective as supplements in milk for passive protection. Furthermore, these monoclonal antibodies were useful for boosting the efficacy of TGEV-neutralizing colostrum, which by itself was ineffective. These results show that other antigenic sites, different from the 4-well characterized epitopes on the S glycoprotein of TGEV, also are important for passive protection.     

雌二醇多克隆抗体的制备与鉴定

制备雌二醇完全抗原,并通过免疫家兔得到多克隆抗体,为下一步制备雌二醇单克隆抗体和雌二醇检测ELISA试剂盒奠定基础。试验以牛血清白蛋白(BSA)、卵清蛋白(OVA)为载体,采用碳化二亚胺法,合成了雌二醇(E2)的2种免疫偶合物:免疫原E2-BSA和包被原E2-OVA;通过紫外光谱定性证明偶合物偶联成功与否,并对偶合物的蛋白含量、结合比进行测定并以免疫原E2-BSA免疫家兔,制备多克隆抗体,用包被原E2-OVA进行ELISA,对多克隆抗体特异性及效价进行检测。结果表明成功合成了雌二醇人工抗原即免疫原E2-BSA和包被原E2-OVA,二者的蛋白浓度分别为5.455和7.533mg/mL,结合比分别为7:1和8:1;制备的多克隆抗体特异性好,血清效价为1:106。     

兔自家大肠杆菌高免卵黄抗体的研制与应用

采用灭活菌苗进行基础免疫、加强免疫,灭活油乳剂疫苗进行超强化免疫和维持免疫程序。通过微量凝集试验方法,研究了鸡抗兔大肠杆菌高免卵黄抗体的消长规律及临床应用。结果表明,超强化免疫5周后,抗体效价达到高峰,9周后抗体效价逐渐下降。同一剂量卵黄抗体效价达26时,肌注2 mL/kg体重,连用3 d对兔大肠杆菌感染的治愈率达90%。成品卵黄抗体的效价应达到26、安全性检验、无菌检验阴性方可用于临床治疗。卵黄抗体在血清中维持高浓度可持续1周,然后逐渐下降。在冷冻环境下可保存1年,其抗体效价保持在有效水平。与头孢噻肟进行的对比田间治疗试验,卵黄抗体的治愈率显著高于头孢噻肟。     

Duration of protection of calves against rhinovirus challenge exposure by infectious bovine rhinotracheitis virus-induced interferon in nasal secretions

Six calves inoculated intranasally with a vaccinal strain of infectious bovine rhinotracheitis (IBR) virus and 6 control calves were given a placebo. All calves were subsequently challenge exposed (by aerosol) with rhinovirus--3 of the calves from each group at 2 days after they were inoculated with IBR virus or with placebo and the remaining calves at 6 days. Nasal excretion of viruses, interferon (IFN) concentrations in nasal secretions (NS), and neutralizing antibody in sera and NS were determined. All calves given the vaccinal IBR virus subsequently had IFN in their NS. Interferon was detected as early as 1 day, reached maximal titers at 2 to 4 days, and persisted in individual calves for 5 to 10 days after inoculation. Rhinovirus shedding was not detected from IBR virus-inoculated calves whose NS contained both rhinovirus antibody and IFN at the time of challenge exposure; such calves were protected at either 2 or 6 days after IBR virus inoculation. The outcome of rhinovirus challenge exposure of calves whose NS contained IFN, but not rhinovirus antibody, varied with the day of challenge exposure. Rhinovirus excretion was detected from 2 of these calves challenge exposed 2 days after IBR virus inoculation, but was not detected from a calf challenge exposed 6 days after inoculation. However, while IFN was present in NS from the former 2 calves, rhinovirus shedding was markedly reduced as compared with that from control calves without IFN or NS antibody at the time of challenge exposure. Consistent relationship was not observed between the rhinovirus neutralizing antibody titer of calves' sera and NS. The antibody titer of NS more closely correlated with protective immunity to rhinovirus infection than did the serum antibody titer.     

酸处理的沙门氏菌作为异种细菌抗原的免疫载体

用酸对鼠伤寒沙门氏菌进行处理,制成裸细菌,作为空肠弯曲菌共同抗原(CA)的载体.用这种裸细菌和CA的复合物免疫小鼠,以间接ELISA和琼脂糖凝胶扩散沉淀反应检测免疫反应的变化及血清效价.结果表明,用这种方法免疫的小鼠,其血清抗体的ELISA滴度与对照组(福氏完全佐剂+CA组,CA组)相差不显著,但能与可溶性CA形成沉淀线,并含有IgM和IgG两类抗体;而对照组则不与CA出现沉淀反应,且只含IgM一类抗体.这表明,酸处理的沙门氏菌作为异种细菌抗原的载体,在改变机体对某种抗原的免疫类型上具有一定的作用.     

Development of an enzyme-linked immunosorbent assay for Bordetella avium

A Bordetella avium enzyme-linked immunosorbent assay (ELISA) was developed to detect serum antibodies in 1-day-old poults, experimentally infected turkeys, and naturally infected turkeys. The optimized procedure included use of a suspension of whole bacteria coated onto plastic microtiter plates, a 1:200 serum dilution, a 1:3200 dilution of commercially available goat anti-turkey IgG (heavy and light chain) conjugated with horseradish peroxidase, and 0.04% orthophenylenediamine as substrate. A sample/negative (S/N) ratio method of analysis was used to estimate antibody titer from absorbance values. The regression equation used to estimate antibody titers was derived from the testing of naturally infected turkey sera. The equation was derived by plotting the log10 titer of the sera against the S/N ratio at a 1:200 serum dilution. The ELISA was an effective method for detecting antibody to B. avium, and the procedure should prove useful for laboratories equipped for high-volume ELISA testing.     

出血性大肠杆菌O157 Eae-Stx1/2B融合蛋白的免疫学特性

为了研究重构融合蛋白Eae-Stx1/2B免疫特性及对出血性大肠杆菌感染的免疫保护作用,进行了细胞粘附抑制试验和免疫保护试验。结果证明Eae-Stx1/2B融合蛋白免疫血清在体外能降低出血性大肠杆菌对HEp-2细胞的粘附作用,这一特性在出血性大肠杆菌疫苗设计中具有重要价值。以纯化的融合蛋白Eae-Stx1/2B为抗原对小鼠进行腹腔注射免疫和滴鼻免疫,ELISA检测结果显示,2次免疫后7 d,免疫组抗Eae-Stx1/2B融合蛋白和抗出血性大肠杆菌O157血清IgG抗体显著高于未免疫组;用出血性大肠杆菌O157强毒株EDL933攻击免疫小鼠,免疫组小鼠排菌时间显著缩短,免疫组小鼠存活率均高于未免疫组,免疫组小鼠体质量恢复增长较快。上述试验结果表明,Eae-Stx1/2B融合蛋白对出血性大肠杆菌感染有保护作用。     

禽流感血清抗体与卵黄抗体消长规律比较研究

应用禽流感病毒二价（H5、H9）油乳剂灭活疫苗免疫160日龄非免疫蛋鸡，免疫接种后分别于第0、7、14、21、28、35 d采集相应的鸡蛋和血清，采用血凝抑制(HI)试验监测血清及卵黄中H5、H9抗体的消长规律，结果表明：H5、H9血清抗体于免疫后7 d时开始产生，14 d时到中等水平，21 d时达到最高值，一直维持到35 d之后；而H5、H9卵黄抗体与血清抗体水平相比相对滞后7 d左右；卵黄抗体与血清抗体在28 d时均达到高峰值并一直维持到35 d之后，28 d后血清和卵黄抗体达到相同水平。本研究为临床上以禽流感卵黄抗体检测替代血清抗体检测及以后禽流感高免卵黄抗体的研究提供了依据和数据。     

抑制素α主动和被动免疫对哈萨克羊生殖激素含量的影响

为了研究抑制素α（INHα）主动和被动免疫对哈萨克羊生殖激素含量的影响，本试验在对抑制素α重组质粒表达菌株进行诱导表达的基础上，将经纯化、鉴定的抑制素α重组蛋白免疫接种新疆双峰骆驼，制备驼抗抑制素α多克隆抗体，并对其进行纯化，检测抗体效价，验证抗体的特异性。之后选择3~5岁、发情时间相近并处于间情期的45只成年哈萨克羊随机分为3组，分别作为抑制素α多克隆抗体免疫组（A组）、抑制素α重组蛋白免疫组（B组）及对照组（C组），每隔10 d连续进行3次加强免疫，应用ELISA法检测在绵羊繁殖活动中具有重要功能的5种生殖激素：促卵泡素（FSH）、促黄体素（LH）、孕酮素（P₄）、雌激素（E₂）、抑制素（INH），并检测血液生化指标。结果显示，IPTG的最佳诱导浓度是0.6 mmol/L，在4 h时诱导出产量较高的抑制素α包涵体蛋白，纯化后的抑制素α重组蛋白纯度较高，并具有较好的免疫原性，经进一步验证发现，制备的抑制素α抗体效价为1：512 000，该抗体可与抑制素α重组蛋白特异性结合。说明成功制备了具有免疫原性的抑制素α重组蛋白和高效价的驼抗抑制素α多克隆抗体。免疫后A组LH、P₄含量和B组FSH、LH、P₄、E₂、INH含量与C组相比差异不显著（P>0.05），而A组FSH含量和E₂含量显著高于C组（P<0.05），INH含量显著低于C组（P<0.05）。通过血液生化指标检测发现，抑制素α蛋白和抑制素α抗血清两种免疫制剂免疫后，试验动物均没有出现不良症状。说明两种抑制素α抗原均可对哈萨克羊血液生殖激素的分泌产生良好效果，相比之下抑制素α抗血清免疫效果更佳。