酶免疫法测定鸡血清中磺胺喹恶啉的残留

磺胺喹恶啉（ｓｕｌｆａｑｕｉｎｏｘａｌｉｎｅ，ＳＱ）作为一种常见的抗球虫药，常与氨丙啉等混合使用。但该药物有一定的毒性［１］。美国ＦＤＡ规定，鸡的ＳＱ休药期为１０ｄ，并规定母鸡在产蛋期内禁用ＳＱ。我国规定磺胺类药物的总残留量不得超过３００ｎｇ／ｇ，但尚未有ＳＱ在鸡体最高残留限量及休药期的规定。测定磺胺喹恶啉残留的方法有高压液相法［２］、化学发光法［３］等。酶免疫法测定药物残留具有灵敏、快速、方便等优点，磺胺二甲嘧啶等药物用该法已有成功测定的报道［４］，快速测定牛奶中磺胺二甲嘧啶的酶免疫测定试剂盒…     

单克隆抗体介导抗原捕获酶联免疫吸附试验鉴定传染性支气…

作者建立了一种抗原捕获酶联免疫吸附试验（Ｃ－ＥＬＩＳＡ）。用对各血清型Ｓ１糖蛋白有特异性的单克隆抗体（ＭＡｂ），对阿肯色（ＡｒＫ）、康涅狄格（Ｃｏｎｎ）、马萨诸塞（Ｍａｓｓ）等血清型的传染性支气管炎病毒（ＩＢＶ）进行了检测和鉴定。当Ｓ１特异性单克隆抗体、抗原、鸡血清的血清型相同时，试验（采用双抗体夹心法）结果最为理想。而用群特异性（Ｍ糖蛋白特异性）ＭＡｂ捕获抗原。抗原和异源鸡血清之间有不同程度的交     

抗绵羊肺炎支原体单克隆抗体杂交瘤细胞株的建立

用绵羊肺炎支原体（ＭＯ）抗原免疫的ＢＡＬＢ／Ｃ小鼠，取血清抗体阳性免疫鼠的脾细胞与ＳＰ２／Ｏ细胞在聚乙二醇作用下进行融合，产生杂交瘤细胞、用间接ＥＬＩＳＡ法对杂交瘤细胞生长孔上清液进行检测，筛选阳性杂交瘤细胞株．以有限稀释法克隆１～２次，得到６株分泌抗绵羊肺炎支原体抗原的单克隆抗体（ＭｃＡｂ）杂交瘤细胞株．选择抗体分泌较高的１４Ａ６、７Ａ２７、Ｂ３８进行了较详细的研究，结果表明，其核内染色体数为９４～９６．抗体属性是：１株为ＩｇＧ２ａ、２株为ＩｇＧ１．用间接ＥＬＩＳＡ法对３株ＭｃＡｂ作符异性分析、该ＭｃＡｂ只特异地与绵羊肺炎支原体抗原发生反应，而不与猪肺炎支原体、山羊无乳症支原体抗原发生反应．将杂交瘤细胞注射ＢＡＬＢ／Ｃ小鼠诱生腹水．其抗体效价提高１００倍．杂交瘤细胞在液氮中保存１～２年后复苏仍能稳定地分泌抗绵羊肺炎支原体ＭｃＡｂ。     

诊断B组轮状病毒的三种酶联免疫方法的建立

用从当地分离鉴定的山羊Ｂ组轮状病毒ＫＢ－６３毒株，在剥夺初乳的山羊羔体内传代繁殖病毒，制备抗原，建立了酶联免疫吸附试验（ＥＬＩＳＡ）、斑点酶联免疫吸附试验（Ｄｏｔ－ＥＬＩＳＡ）及生物素－亲和素系统酶联免疫吸附试验（ＢＡＢ－ＥＬＩＳＡ）。这三种方法经阻断试验、重复性试验以及交叉试验表明，其特异性、重复性好。以双盲法将此三种方法与对流免疫电泳法（ＣＩＥ）和聚丙烯酰胺凝胶电泳法（ＰＡＧＥ）作了对比研究，     

分泌抗细粒棘球蚴原头节可溶性抗原单克隆抗体杂交瘤细胞株的建立

用ＳＰ２／０骨髓瘤细胞与以羊源细粒棘球蚴原头节可溶性抗原免疫的Ｂａｌｌ／ｃ鼠脾细胞融合，经克隆和筛选后获得８株分泌抗ＳＰＡ的杂交瘤细胞株，即３Ａ４，１ＩＣ２，２Ｂ３，２Ｃ２，２Ｅ３，３Ｅ６，３Ｆ１０和２Ｃ２。亚类鉴定为ＩｇＧ１（３Ａ４，３Ｅ６，１Ｃ２，３Ｆ１０和３Ｃ２），ＩｇＧ２ｂ（２Ｃ２），ＩｇＧ３（２Ｅ３），ＩｇＧ总（２Ｂ３）。免疫双扩散法显示２Ｃ２腹水及２Ｂ３，２Ｃ２，２Ｅ３培养上清粗提物与     

斑点免疫金银染色法检测鸡传染性支气管炎病毒抗原的研究

用建立的斑点免疫金银染色（Ｄｏｔ－ＩＧＳＳ）法检测鸡传染性支气管炎病毒（ＩＢＶ）抗原,确定兔抗ＩＢＶ血清工作浓度为１：４００,ＳＰＡ－胶体金探针的工作浓度为１：８０,该法对纯化ＩＢＶ抗原的最低检出量为０．４３１４ｎｇ／点,用Ｄｏｔ－ＩＧＳＳ与斑点酶联免疫吸附试验（Ｄｏｔ－ＥＬＩＳＡ）同时检测１５只人工感染ＩＢＶ鸡的气管,肺,肾病科,ＩＢＶ阳性检出率均为１００％,对３２份疑似ＩＢＶ感染鸡病料检测,Ｉ     

抗奶牛衣原体单克隆抗体杂交瘤细胞株的建立

将奶牛衣原体抗原免疫的 Ｂ Ａ Ｌ Ｂ／ｃ 小鼠脾细胞与 Ｓ Ｐ２／ Ｏ 细胞在聚乙二醇作用下融合, 用间接 Ｅ Ｌ Ｉ Ｓ Ａ 试验筛选, 以有限稀释法克隆３ 次, 得到６ 株分泌抗奶牛衣原体单克隆抗体（ Ｍｃ Ａｂ） , 选择抗体分泌较高的 Ｇ２ 、 Ｆ２３ 、 Ａ 株进行详细的研究, 结果表明, 其核内染色体数为９０３５ 、９２１４ 、９４４２ ; 抗体属性为 Ｉｇ Ｇ２ａ 、 Ｉｇ Ｇ１ 、 Ｉｇ Ｍ, 用交叉 Ｅ Ｌ Ｉ Ｓ Ａ 法对３ 株 Ｍｃ Ａｂ 作特异性分析, 该 Ｍｃ Ａｂ 只与奶牛衣原体抗原, 猪衣原体抗原、羊衣原体抗原发生反应,而不与沙眼衣原体抗原、伪狂犬病抗原、布鲁氏杆菌抗原发生反应。     

传染性支气管炎病毒S1基因DNA免疫质粒的构建及其免疫原性

为探讨ＤＮＡ免疫防制传染性支气管炎（ＩＢ）的可能性，以克隆的Ｍａｓｓｃｈｕｓｅｔｅ型类Ｍ４１地方分离ＱＤ株Ｓ１基因为免疫原基因，引入终止密码后插入ＳＶ４０启动子、增强子下游和ＳＶ４０ｐｏｌｙＡ信号上游，构建了Ｓ１基因ＤＮＡ免疫表达质粒ｐＳＶＱＤＳ１。将大量扩增并经聚乙二醇纯化的ｐＳＶＱＤＳ１溶于ＰＢＳ，以３００μｇ／只的剂量肌肉注射免疫小鼠，于４周后采集分离免疫小鼠血清进行鸡胚病毒中和试验。结果表明，ｐＳＶＱＤＳ１能够诱导产生针对传染性支气管炎病毒（ＩＢＶ）Ｍ４１株的中和抗体，具有良好的免疫原性。     

猪水泡病病毒抗独特型抗体的研究—Ab2的制备与鉴定

用猪水泡病病毒中和单抗２Ｂ６和４Ｈ９免疫家兔，制备出ＳＶＤＶ兔抗独特型抗体。经亲和层析纯化，用ＥＬＩＳＡ鉴定性质，结果表明，所制备的Ａｂ２能竞争抑制Ａｂ１与ＳＶＤＶ抗原结合，同时，Ａｂ２与Ａｂ１的结合可被病毒抗原阻断，表明所制备的Ａｂ２具有ＳＶＤＶ抗原内影像关系。     

分泌抗赭曲霉素A单克隆抗体杂交瘤细胞株的建立

用赭曲霉素Ａ（ＯＡ）－ＢＳＡ合成抗原免疫Ｂａｌｂ／ｃ小鼠，取免疫鼠脾细胞与ｓｐ２／０细胞融合，通过克隆和ＥＬＩＳＡ法筛选，建立三株泌抗ＯＡ单克隆抗体杂交瘤细胞株（３Ｃ１２，１Ｄ１２和２Ｇ７）。用间接ＥＬＩＳＡ法测定，细胞上清液抗体效价为１２８＾×（３Ｃ１２）和６４＾×（１Ｄ１２和２Ｇ７）腹水抗体效价为１０＾－７（３Ｃ１２，１Ｄ１２）和１０＾－６（２Ｇ７）。三株单抗（ＭｃＡｂ）均属ＩｇＧ类，分泌抗体     

Taeniacidal efficacy of SQ 21,704 in dogs by various types of oral administration and in comparison with niclosamide and bunamidine hydrochloride

The streptothrinin antibiotics SQ 21,704 was evaluated against naturally occurring Taenia pisiformis and Dipylidium caninum infections in dogs when they were given at a dose level of 37.5 mg/kg of body weight in four different rations: loaf-type canned meat; chunk-type canned meat; dry (gravy-type) meal; and dry (pelleted) meal. The SQ 21,704 was 100% efficacious against both T pisiformis and D caninum infections when given with the chunk, gravy, and pelleted rations. When given with the loaf-type canned meat, it was 100% effective against T pisiformis and 60% efficacious against D caninum. The SQ 21,704 was effective against both tapeworm species when given orally as a liquid at a dose level of 37.5 mg/kg, formulated as an aqueous suspension containing 94 mg of activity per milliter. The SQ 21,704 was also tested in dogs when given orally in gelatin capsules at a dose level of 37.5 mg/kg without fasting, and was 100% efficacious against both tapeworm species. The results of a comparative taeniacidal study demonstrated that SQ 21,704 was 100% effective in removing both T pisiformis and D caninum when administered orally at a dose level of 37.5 mg/kg, whereas niclosamide and bunamidine hydrochloride were only partially effective at their recommended dose levels. One of five dogs treated with niclosamide at a dose level of 157 mg/kg was positive at necropsy, giving an orally efficacy of 80%. Three of five dogs treated with bunamidine hydrochloride at a dose level of 49 mg/kg were positive at necropsy, giving an overall efficacy of 40%.     

Reproductive safety of vaccination with Vista 5 L5 SQ near breeding time as determined by the effect on conception rates

Replacement heifers (N=799; 10 to 13 months of age) were vaccinated with Vista 5 L5 SQ (Intervet; a reconstituted vaccine-bacterin product containing modified-live cultures of infectious bovine rhinotracheitis [IBR] virus, bovine viral diarrhea virus [BVDV; types 1 and 2], parainfluenza-3 virus, and bovine respiratory syncytial virus and inactivated cultures of Leptospira serovars canicola, grippotyphosa, hardjo, icterohaemorrhagiae, and pomona with a proprietary adjuvant) at either 40 plus/minus 5 days (control; n=399) or 3 days (test; n=400) before peak breeding day. By 40 plus/minus 5 days before peak breeding day, heifers in both groups had greater average titers to IBR, BVDV types 1 and 2, and four of the five Leptospira antigens assessed as compared with prevaccination titers on day -90 plus/minus 25 days. Conception rates were not affected by treatment. This study suggests that conception rates will not differ between heifers vaccinated with Vista 5 L5 SQ 3 days before breeding and those vaccinated approximately 40 days before breeding.     

Decreasing profile of residual sulphaquinoxaline in eggs

1. Sulphaquinoxaline (SQ) was added to the diet of laying hens at 200 mg/kg for 7 successive days. Contents (mg/kg) of SQ in albumen and egg yolk of eggs laid after drug withdrawal were determined by high pressure liquid chromatography (HPLC). The contents in the whole egg were calculated taking into consideration the respective weights of albumen and egg yolk. 2. A time-lag in the initiation of decrease of SQ from eggs after the withdrawal of dietary SQ was observed. 3. The decreasing pattern from whole egg could be well described by the following equation with a time-lag of 1.0 d, y = 2.07e-0.5620(t-1.0), where y is the SQ content in whole egg, t is time (d) after the withdrawal of dietary SQ and a constant of 2.07 is the SQ content in whole egg laid at the withdrawal. 4. Biological half-life of SQ in the whole egg was estimated to be 1.23 d. 5. From the above equation, SQ residue of whole egg laid at 9th d after withdrawal will be below the detection limit of 0.01 mg/kg.     

Critical evaluation of taeniacidal antibiotic S15-1 (SQ 21, 704) for removal of natural tapeworm infections in dogs and cats

The new taeniacidal antibiotic S15-1 (SQ 21,704) was evaluated against naturally occuring infections of Taenia pisiformis in 53 dogs, Dipylidium caninum in 35 dogs, T taeniaformis in 18 cats, and D caninum in 33 cats. It all instances, the compound was administered in gelatine capsules in a single oral dose. The doses tested were between and 200 mg/kg of body weight in dogs and between 15 and 45 mg/kg in cats. In dogs, doses of 25 mg/kg and greater were 100% effective against T pisiformis, whereas a dose of 50 mg/kg was necessary to clear D caninum. In cats, a single oral dose of 22.5 mg/kg was 100% efficacious against T taeniaeformis, and a single dose of 45 mg/kg (the largest dose tested) clearly seven of eight cats of D caninum. The efficacy was limited to tapeworms only; there was no efficacy against nematodes. The antibiotic was well tolerated by both species with no drug-related vomiting or other side-effects observed.     

Tissue concentrations of sulphaquinoxaline administered in the food of laying hens

1. Sulphaquinoxaline (SQ) was fed to laying hens at a dietary level of 400 mg/kg for 3 successive days. SQ contents (mg/kg) in the blood, kidney, liver, ovary, muscle (thigh) and adipose tissue collected on 1, 2, and 3 d after the start of feeding were determined by HPLC. The relationship between the SQ content in the tissues and times (d) of SQ feeding was analyzed statistically. 2. Dietary SQ was distributed throughout the whole body. 3. Contents in tissues, except the kidney, had already reached a plateau by day 1 after the start of administration whereas in the kidney it increased linearly throughout the 3 days. 4. The plateau values of SQ in the tissues were much greater than those of sulphamonomethoxine and sulphadimethoxine.     

应用mRNA差异显示技术筛选绵羊皮肤毛囊差异表达序列标签

为寻找与绵羊羊毛生长相关的基因,应用mRNA差异显示技术(DD-PCR)筛选拔毛皮肤与正常皮肤表达水平有差异的cDNA,用Northern blot证实差异显示基因片段,对差异基因进行测序和同源比较。经差异显示分析,共找到差异表达的序列片段21个;经2次扩增、克隆、Northern Blot分析,最终确定2个差异条带SQ70和SQ75。对这两个差异条带进行测序及生物信息学分析,发现SQ70是绵羊皮肤中与组织修复相关的EST片段,而SQ75是绵羊皮肤或者是毛囊中高表达的未知EST片段。以上结果表明,SQ70和SQ75可能是与绵羊羊毛生长密切相关的基因表达产物,对于这两个ESTs具体的生物学功能有待于进一步研究。     

Expression analyses of candidate genes related to meat quality traits in squabs from two breeds of meat‐type pigeon

In this study, meat quality traits were compared between squabs from two pigeon breeds: one Chinese indigenous breed, the Shiqi (SQ) meat‐type pigeon, and an imported breed, the white king (WK) meat‐type pigeon. Breed differences were detected in the content of intramuscular fat (IMF) in the breast muscle. SQ squabs had significantly higher IMF content than the WK birds. The shear force value (an objective measure of meat tenderness) of SQ birds was also relatively lower than that of the WK squabs. Further analysis of fatty acids profile revealed that SQ squabs exhibited significant advantage in the synthesis of polyunsaturated fatty acids, while WK squabs were significantly higher in the sum of monounsaturated fatty acids. Breast muscle in the SQ squabs was also significantly higher in the ratio of polyunsaturated fatty acids to saturated fatty acids, as well as the sum of omega 6 fatty acids. Variability of expression levels of functional genes in relation to fat accumulation and meat tenderness was analysed by qRT‐PCR. Gene expression analyses showed that the hepatic expression of LPL (lipoprotein lipase), FABP4 (fatty acid‐binding protein 4), and CAPN2 (calpain‐2) were significantly higher in the SQ squabs. In the breast muscle tissue, the FABP3 (fatty acid‐binding protein 3) and CAPN2mRNA abundance was significantly higher in SQ squabs. Our results suggested that these differentially expressed genes might be candidate genes used in the programmes of targeted selection for squabs with higher IMF content, tender meat, and more favourable fatty acids composition.     

Coccidiosis: a radiological study of sulphaquinoxaline distribution in infected and uninfected chickens

Using scintillation counting and autoradiographical techniques, the whole-body distribution in week-old uninfected chickens of the anticoccidial agent sulphaquinoxaline (SQ) labelled with ³⁵S was established at various time intervals after a single oral dose either alone or following continuous in-feed medication with unlabelled SQ, and after a single intravenous dose. The distribution was also established in chickens infected with the coccidia Eimeria acervulina or E. tenella , after a single oral dose of radiolabelled SQ administered either alone or following continuous in-feed medication with unlabelled SQ, as for uninfected chicks. In all uninfected chicks, SQ was rapidly absorbed from the gut and was distributed to all tissues. It appeared at high concentrations in the bile and kidneys 0.5 h after dosing. In chickens that had previously received unlabelled SQ in the diet, a radiolabelled dose maintained steadier tissue concentrations than the sharp rise and fall detected after a single oral dose. Intravenous dosing of uninfected chicks showed that SQ was secreted by the crop, gizzard and caecal epithelia into their lumina. Infection with E. acervulina or E. tenella coincided with an overall 3.5-fold sustained increase of SQ concentration in chick tissues. An updated hypothesis including these new observations for the anticoccidial mode of action of SQ in chickens is expounded.     

Tissue residues of some sulphonamides in normal and Eimeria stiedai infected rabbits.

Tissue residues of sulphadiazine (SDZ), sulphadimidine (SDD) and sulphquinoxaline (SQ) were studied in healthy and E. stiedai infected rabbits following oral administration of 0.5 g/l drinking water for 5 days. The solid-phase extraction and HPLC was used to determine the concentration of the three sulphonamides in a single tissue sample. SDZ was detected in the liver and kidney in concentrations below the tolerance levels at day 5 and no residues could be detected at day 7 after drug withdrawal. SDD and SQ were detected in all of the tested organs of healthy rabbits up to day 5, where the highest concentration was reported in the liver (0.08 +/- 0.02 and 0.09 +/- 0.02 g/g respectively). In infected rabbits, the three sulphonamides were detected up to day 7 in concentrations higher than the tolerance limits (> 0.1 g/g) in the liver and kidney and lower levels in other tissues. A withdrawal period of 4 days for SDZ and 5 days for SDD and SQ in healthy rabbits and 7 days for SDZ and 8 days for SDD and SQ in E. stiedai infected rabbits is suggested.     

Vaccination of day-old broiler chicks against Newcastle disease and infectious bursal disease using commercial live and/or inactivated vaccines

Day-old broilers were administered live and/or inactivated vaccines to assess vaccine efficacy against challenge with Newcastle disease (ND) and infectious bursal disease (IBD). Chicks were from commercial breeder pullets vaccinated against ND and IBD using several live vaccine primers followed by an inactivated ND-IBD vaccine at 18 weeks. The most efficacious initial ND-IBD vaccination program was live ND virus by eye drop and live IBD vaccine injected subcutaneously (SQ) followed 2 hours later with inactivated ND-IBD vaccine SQ. The next two most efficacious programs were live vaccine alone and the inactivated vaccine only. Inactivated vaccine given SQ had no adverse effect on live IBD vaccine given 2 hours earlier in a similar site. Administration of inactivated vaccine by vent was not as efficacious as administration SQ. A booster of a second live ND-IBD vaccine drinking water at 18 days significantly increased levels of circulating antibody, regardless of the initial vaccination program.