不同生理时期不同水平大豆抗营养因子对生长蛋鸡器官重的影响

不同生理时期不同水平大豆抗营养因子对生长蛋鸡器官重的影响ＥＦＦＥＣＴＳＯＦＴＨＥＤＩＥＴＳＣＯＮＴＡＩＮＩＮＧＶＡＲＩＮＧＬＥＶＥＬＳＯＦＲＡＷＳＯＹＢＥＡＮＯＮＯＲＧＡＮＷＥＩＧＨＴＳＯＦＰＵＬＬＥＴＳＤＵＲＩＮＧＤＩＦＦＥＲＥＮＴＧＲＯＷＩＮＧＰ...     

鸡传染性支气管炎病毒S1基因在昆虫细胞中的高效表达

将鸡传染性支气管炎病毒（ Ｉ Ｂ Ｖ） Ｓ１ 基因 ｃ Ｄ Ｎ Ａ 克隆至含有起始密码 Ａ Ｔ Ｇ 的转移载体 ｐ Ｓ Ｘ Ｉ Ｖ Ｖ Ｉ＋ Ｘ３／４,构建成重组转移质粒 ｐ Ｓ Ｘ Ｉ Ｖ Ｖ Ｉ＋ Ｘ３／４ Ｓ１． Ｈｏｌｔｅ,再与粉纹夜蛾核型多角体病毒 Ｔｎ Ｎ Ｐ Ｖ Ｓ Ｖ Ｉ－ Ｇ Ｄ Ｎ Ａ（ Ｏ Ｃ Ｃ－ , ｇａｌ＋ ）共转染 草地夜蛾（ Ｓｆ９） 细胞, 经空斑纯化得 到已插入 Ｓ１ 基 因并能形成多角体 的重组病毒 Ｔｎ Ｎ Ｐ Ｖ（ Ｘ３／４） Ｓ１． Ｈｏｌｔｅ Ｏ Ｃ Ｃ＋ 。以重组毒株感染 Ｔｎ５ Ｂ１ 细胞,在不同时间收取感染了病毒的细胞进行 Ｓ Ｄ Ｓ Ｐ Ａ Ｇ Ｅ 与 Ｗ ｅｓｔｅｒｎ 印迹检测。 Ｔｎ Ｎ Ｐ Ｖ（ Ｘ３／４） Ｓ１． Ｈｏｌｔｅ Ｏ Ｃ Ｃ＋ 在感染的细胞中高效表达了 Ｓ１ 基因,表达产物为约 １００ ０００ 的融合蛋白,与预计的表达修饰后的产物大小相符,推测此蛋白已糖基化。 Ｓ Ｄ Ｓ Ｐ Ａ Ｇ Ｅ凝胶薄层色谱分析结果显示,感染病毒后 ４８、７２、９６ ｈ, Ｓ１ 蛋白分别占细胞内总蛋白量的 ２８７％ 、３５８％ 、３７１％ 。     

中国饲料成分及营养价值表

ＴＡＢＬＥＳＯＦＦＥＥＤＣＯＭＰＯＳＩＴＩＯＮＡＮＤＮＵＴＲＩＴＩＶＥＶＡＬＵＥＳＩＮＣＨＩＮＡ表 4 .矿物质及维生素含量ＭｉｎｅｒａｌｓａｎｄＶｉｔａｍｉｎｓ序号 中国饲料号ＣＦＮ 饲料名称ＦｅｅｄＮａｍｅ 钠Ｎａ( % ) 氯Ｃｌ( % ) 镁Ｍｇ( % ) 钾Ｋ ( % ) 铁Ｆｅ(ｍｇ/ｋｇ)铜Ｃｕ(ｍｇ/ｋｇ)锰Ｍｎ(ｍｇ/ｋｇ)1 4- 0 7- 0 2 78玉米ｃｏｒｎｇｒａｉｎ 0 .0 1 0 .0 4 0 .1 1 0 .2 936 3.4 5.82 4 - 0 7- 0 2 88玉米ｃｏｒｎｇｒａｉｎ 0 .0 1 0 .0 4 0 .1 1 0 .2 936 3.4 5.834 - 0 7- 0 2 79玉米ｃｏｒｎｇｒａｉｎ 0 .0 …     

马立克氏病病毒囊膜糖蛋白B抗原Ⅰ型特异性和群共同性决定簇的单克隆抗体

用能表达马立克氏病病毒（ＭＤＶ）糖蛋白Ｂ（ｇＢ）的重组杆状病毒感染的Ｓｆ９细胞免疫小鼠，制备针对ＭＤＶｇＢ的单克隆抗体。以Ⅰ型马立克氏病病毒ＧＡ株感染的鸡胚成纤维细胞作为检测抗原，同时以免疫荧光试验（ＩＦＡ）和酶联免疫吸附试验（ＥＬＩＳＡ）来筛选杂交瘤细胞，结果获得了ＩＦＡ和ＥＬＩＳＡ均为阳性的２株单克隆抗体细胞株，定名为ＢＡ４和ＢＤ８。在ＩＦＡ和免疫沉淀试验中，单抗ＢＤ８与Ⅰ、Ⅱ、Ⅲ型ＭＤＶ均呈阳性反应；单抗ＢＡ４只对Ⅰ型ＭＤＶ（包括ＣＶＩ９８８疫苗株）呈阳性反应。免疫沉淀反应进一步确证２株单抗识别的是ＭＤＶ糖蛋白Ｂ抗原。     

pCMV—TNFα的构建及其在动物细胞中的表达

以Ｘｂａ Ｉ和Ｅｃｏ ＲＶ双酶消化ｐＢＴ１获得肿瘤坏死因子α（ＴＮＦα）ｃＤＮＡ基因片段，以Ｘｂａ Ｉ和Ｓｍａ Ｉ酶切点定向克隆入质粒载体ｐＣＭＶ４，构建了人ＴＮＦα重组质粒ｐＣＭＶ－ＴＮＦα。采用ＤＮＡ－磷酸钙共沉淀法转染ＣＯＳ７和ＮＩＨ３Ｔ３细胞，收集转染后不同时间的细胞培养上清，检测ＴＮＦα的表达水平和生物学活性。ＥＬＩＳＡ检测表明，两种细胞转染后，其培养上清中均有ＴＮＦα抗原存在，而载体等     

猪生殖—呼吸道综合征病毒CH—1a株N基因的原核表达

将ＰＲＲＳＶＣＨ－１ａ株Ｎ基因用ＥｃｏＲＩ和ＰｓｔＩ双酶切从重组质粒ｐＵＣ１８－ＯＲＦ７切下后,插入到原核表达载体ｐＢＶ２２０的ＰＲ,ＰＬ启动子下游,得到重组表达质粒,ｐＢＶ２２０－ＯＲＦ７,转化了ｐＢＶ２２０－ＯＲＦ７的大肠杆菌ＪＭ８３经诱导培养后,用ＳＤＳ－ＰＡＧＥ和Ｗｅｓｔｅｒｎｂｌｏｔ检测表达产物,结果表明ＰＲＲＳＶＣＨ－１ａ株的Ｎ基因在原核载体上得到高效表达,表达产物占菌体总蛋白的１５．     

FMDV在BHK21细胞扩增后FMDV 140 S抗原的测定

本文报告了用蔗糖密度梯度离心和光密度分析相结合口蹄疫病毒（ＦＭＤＶ）在单层ＢＨＫ２１细胞中扩增后，其中ＦＭＤＶ １４０ Ｓ抗原浓度，并同时分析了经ＢＥＩ（二乙烯亚胺）灭活后，这种抗原含量的改变。结果表明，Ｏ型ＦＭＤＶ在单层ＢＨＫ２１细胞中增殖后，其ＦＭＤＶ １４０ Ｓ抗原浓度降至０．９μｇ／ｍｌ。     

不同细胞培养基细胞培养动力学比较试验

应用宜兴赛尔生物化工厂产１９９、Ｆ－１０、１６４０和ＤＭＥＭ细胞培养基与美国Ｓｉｇｍａ公司产１９９、Ｆ－１０、１６４０和ＤＭＥＭ细胞培养基分别配制细胞培养液，培养ＳＰ２／０和Ｖｅｒｏ细胞系及原代鸡胚成纤维细胞（ＣＥＦ），做细胞培养动力学比较试验。比较这两种来源的细胞培养基对细胞生长的形态、分裂速度及鸡马立克氏病毒（Ｍａｒｅｋ＇ｓｄｉｓｅａｓｅｖｉｒｕｓ．ＭＤＶ）疫苗毒株ＨＶＴ－ＦＣ１２６和ＲｉｓｐｅｎｓＣＶＩ９８８在ＣＥＦ单层上增殖量的差异。研究结果表明，四种国产细胞培养基与对应的四种进口培养基对细胞生长及病毒增殖的影响无显著差异     

猪生殖-呼吸道综合征病毒CH-1_株N基因的原核表达

将ＰＲＲＳＶＣＨ１ａ株Ｎ基因用ＥｃｏＲＩ和ＰｓｔＩ双酶切从重组质粒ｐＵＣ１８ＯＲＦ７切下后,插入到原核表达载体ｐＢＶ２２０的ＰＲ、ＰＬ启动子下游,得到重组表达质粒ｐＢＶ２２０ＯＲＦ７。转化了ｐＢＶ２２０ＯＲＦ７的大肠杆菌ＪＭ８３经诱导培养后,用ＳＤＳＰＡＧＥ和Ｗｅｓｔｅｒｎｂｌｏｔ检测表达产物。结果表明ＰＲＲＳＶＣＨ１ａ株的Ｎ基因在原核载体上得到高效表达,表达产物占菌体总蛋白的１５３％。表达蛋白可望成为有价值的ＰＲＲＳ诊断抗原。     

分泌抗IBDV独特型抗体细胞株的建立

应用提取纯化的抗ＩＢＤＶ ＩｇＧ免疫Ｂａｌｂ／ｃ小鼠，其脾细胞与ＳＰ２／Ｏ细胞在ＰＥＧ作用下融合，应用ＥＬＩＳＡ法检测筛选，经有限稀释法克隆２次，获得了２株（２Ｂ６株、５Ｆ４株）分泌抗ＩＢＤＶ独特型抗体的杂交瘤细胞株，并能诱生Ｂａｌｂ／ｃ小鼠产生高效价的含抗ＩＢＤＶ独型抗体腹水。     

The exacerbating effect of infectious bronchitis virus infection on the infectious bursal disease virus-induced suppression of opsonization by Escherichia coil antibody in chickens

Chickens infected with infectious bronchitis virus (IBV) and infectious bursal disease virus (IBDV) commonly develop secondary infection of the respiratory tract with Escherichia coli, resulting in significant economic losses. To understand the host factors that may contribute to the E. coli infection, we investigated macrophage-mediated E. coli phagocytosis, intracellular bacterial killing, and development of opsonizing antibody in previously uninfected chickens and in those infected with IBV, IBDV, and IBDV plus IBV. Macrophages from the peripheral blood and the respiratory tracts of chickens infected with IBV or IBDV plus IBV efficiently performed in vitro phagocytosis of E. coli in the presence of positive-control serum (i.e., E. coli antiserum produced in normal chickens). Those macrophages also had adequate bactericidal activity, indicating that IBV and IBDV infections had not affected their phagocytic activity or bactericidal function. The phagocytic activity of macrophages remained unaffected (P < 0.05) when the positive-control serum was replaced with E. coli antiserum produced in chickens infected with IBV alone. However, when E. coli antisera raised in IBDV-infected and, especially, that produced in IBDV plus IBV-infected chickens were supplemented, the percentage of phagocytosis and number of bacteria ingested per phagocyte were significantly (P < 0.05) less. These results indicate that although IBDV alone has the potential to markedly reduce opsonizing ability of antibody, this effect is significantly (P < 0.05) exacerbated by IBV infection.     

传染性法氏囊病病毒变异E株感染鸡细胞凋亡的研究

研究了传染性法氏囊病病毒（IBDV）变异E株人工感染28日龄SPF雏鸡后鸡法氏囊淋巴细胞的凋亡情况。电镜观察和DNA电泳分析结果表明,IBDV感染后12～48h,雏鸡法氏囊淋巴细胞出现典型细胞凋亡的形态学特征和生化特征;经流式细胞计检测和荧光染色观察,统计学分析表明,IBDV感染后24～48h,雏鸡法氏囊淋巴细胞凋亡数量显著增加（P<0.05或P<0.01）。试验结果揭示IBDV变异E株人工感染可以诱导雏鸡法氏囊淋巴细胞凋亡。     

Infectious bursal disease virus: further characterization with evidence for a single-stranded RNA virus

Infectious bursal disease virus (IBDV) that had been adapted to grow and was then cloned in chick embryo fibroblast (CEF) cell culture was examined for its physicochemical properties, the cellular site of virus replication, and the nature of its viral RNA. The IBDV was an RNA virus, acid-stable, absolutely resistant to chloroform, and moderately thermolabile. It appeared to replicate only in the cytoplasm, as shown by virus-specific antigens restricted to the cytoplasm of infected cells. The viral RNA was composed of single-stranded RNA, as evidenced by flame-red fluorescence on acridine-orange staining and an absence of specific fluorescence in infected cells on immunofluorescent staining with antiserum specific for double-stranded RNA. The IBDV virion had a hexagonal outline with an average diameter of 62 nm and possessed a single layer of capsid composed of hollow capsomeres without envelope. The buoyant density as determined in a continuous sucrose gradient was 1.178 g/cm3. The IBDV was found to possess morphologial and physicochemical properties different from those of any established RNA virus group.     

复方中草药"毒菌杀"对感染IBDV雏鸡白细胞数量及淋巴细胞ANAE+百分率的影响

试验观察了复方中草药“毒菌杀”对感染法氏囊病毒(IBDV)雏鸡的外周血液白细胞及α-醋酸萘酯酶(α-acid naphthyl acetate esterase,ANAE)阳性淋巴细胞百分率的影响。结果发现,“毒菌杀”可以使感染IBD V的雏鸡维持外周血白细胞数量的相对稳定,提高ANAE阳性淋巴细胞的活性水平,具有明显的免疫保护作用。     

Humoral and cell-mediated immune responses in chickens with infectious bursal disease

Primary and secondary immune responses to Newcastle disease virus (NDV) was evaluated in chickens infected with infectious bursal disease virus (IBDV) at one and 28 days of age. The geometric mean primary hemagglutination-inhibition antibody titers (GMT) of chickens infected with IBDV at one day of age was significantly lower (P less than or equal to 0.01) than those infected at 28 days of age. Infection with IBDV had no influence on secondary immune response to NDV. The effect of IBDV infection at one day of age on the cell-mediated immunity of chickens was evaluated by skin allograft acceptance or survival time. There was no significant difference between the percentage of grafts accepted in IBDV infected and noninfected control chickens. However, the mean graft survival time in the IBDV infected chickens was significantly longer (P less than or equal to 0.05) than those in the control group. This suggested a suppression of cell-mediated immunity due to IBDV infection.     

Single and combined infections of specific-pathogen-free chickens with infectious bursal disease virus and an intestinal isolate of reovirus

The susceptibility of 1-day-old and 7-day-old specific-pathogen-free chickens to infection with a virulent strain of infectious bursal disease virus (IBDV) or an intestinal isolate of avian reovirus, or a combination of the two, was investigated. Chickens infected with IBDV and reovirus had more severe pathological lesions than chickens infected with either virus alone, and prior infection with IBDV enhanced the pathogenicity of enteric reovirus. Virus recovery was attempted from bursa, spleen, thymus, liver, intestine, pancreas, cecal tonsils, heart, and tarso-metatarsal tendons. Viruses were recovered from all tissues sampled for either IBDV or reovirus isolation, and indications were that infection with IBDV before infection with reovirus led to longer persistence of reovirus in some tissues. Antibodies to IBDV or reovirus were measured by the virus neutralization test and enzyme-linked immunosorbent assay. Chickens infected with IBDV had lower (P less than 0.05) antibody responses to reovirus than chickens infected with reovirus alone.     

传染性囊病病毒诱导细胞凋亡的初步观察

用１株ＩＢＤＶ强毒株感染易感小鸡，对病鸡法氏囊进行电镜观察及ＤＮＡ电泳分析，直接观察到病鸡法氏囊中Ｂ淋巴细胞凋亡的典型形态学特征和生化变化：染色质凝聚成团，集于核膜旁，胞膜与核膜出现凹陷，细胞拉长变形，最后细胞裂解成由膜包围着的小团，被网状细胞和巨噬细胞吞噬；感染ＩＢＤＶ２４～４８ｈ的法氏囊细胞总ＤＮＡ在电泳谱上呈梯状条带，而从正常的法氏囊提取的总ＤＮＡ在电泳谱上只有１条带。结果表明，ＩＢＤＶ感染小鸡之后，导致了法氏囊中Ｂ淋巴细胞的凋亡。作者据此推断，细胞凋亡是造成Ｂ淋巴细胞数量减少，从而导致小鸡免疫抑制的原因     

An immunoperoxidase monoclonal antibody stain for rapid diagnosis of infectious bursal disease

Cell smears of chicken-embryo-fibroblast (CEF) cultures and bursa of Fabricius from chickens experimentally infected with six different strains of infectious bursal disease virus (IBDV) were examined for the presence of IBDV by the avidin-biotin-peroxidase complex method of immunoperoxidase (IP) staining using a monoclonal antibody specific for IBDV designated BK70. IBDV of different strains and serotypes were readily detected by the IP method in cell smears prepared from infected CEF cultures and from bursas. Bursal cells were positive for IP stain in most of the infected bursas (87.5%), despite their mild IBD lesions. Positive IP staining of bursal smears was well correlated with the recovery of IBDV from the bursas and with IBD lesions in the bursas. IP stain with a monoclonal antibody (BK70) appeared potentially useful for rapid and definitive diagnosis of IBD.     

Protective immunity against infectious bursal disease virus in chickens in the absence of virus-specific antibodies

The role of cell-mediated immunity (CMI) in pathogenesis of infectious bursal disease virus (IBDV) was investigated. One-day-old specific pathogen-free chickens were treated with 3mg of cyclophosphamide (Cy) per chicken for 4 consecutive days and, 3 weeks later, infected with the IBDV-IM strain. Chickens were examined for: (a) mitogenic response of splenocytes to ConA, as an indicator of T-cell functions in vitro, (b) antibody against IBDV by ELISA, (c) IBDV genome in various tissues by RT-PCR and (d) immunological memory. At the time of IBDV infection, Cy-treated chickens had depleted bursal tissue (an avian primary B-cell lymphoid organ), severely compromised antibody-producing ability, but normal T-cell response to ConA. In primary infection, no detectable antibody against IBDV antigen in Cy-treated, IBDV-infected chickens was observed up to 28 days post-infection (PI), while IBDV genome was detected by RT-PCR in spleen, thymus, liver and blood until 10 days PI. Like intact control chickens infected with IBDV, Cy-treated, IBDV-infected chickens suppressed splenocytes responses to ConA from 5 to 10 days PI, suggesting that intact control as well as Cy-treated chickens responded similarly to IBDV infection in the early phase. Following re-infection with IBDV, no detectable secondary antibody response to IBDV as well as IBDV genome in tissues were observed in Cy-treated chickens, while intact control chickens developed vigorous secondary antibody response. Similar to intact control chickens infected with IBDV, Cy-treated chickens after second infection with IBDV did not suppress splenocyte response to ConA. These results suggested that in the absence of detectable anti-IBDV antibodies, protection of Cy-treated chickens from IBDV infection may occur via immunological memory mediated by CMI. We concluded that under normal conditions, IBDV induces a protective antibody response, however, in the absence of antibody, CMI alone is adequate in protecting birds against virulent IBDV.     

Responses of specific-pathogen-free chicks to concomitant infections of reovirus (WVU-2937) and infectious bursal disease virus

Specific-pathogen-free white leghorn chicks concomitantly infected with both infectious bursal disease virus (IBDV) and reovirus (WVU-2937) on day 1 had significantly lower (P less than 0.05) virus-neutralizing- and precipitating-antibody geometric mean titers (GMT) to reovirus than chicks infected with only reovirus on day 1 but had a similar incidence of inflammation of the metatarsal digital flexor tendons. Chicks infected with IBDV on day 7 and reovirus on day 14 had a greater incidence of inflammation of the tarsometatarsal digital flexor tendons and lower neutralizing- and precipitating-antibody GMT to reovirus than chicks infected with only reovirus on day 14. Chicks infected with both viruses on day 1 had a significantly lower (P less than 0.05) neutralizing-antibody GMT to IBDV than chicks infected with only IBDV.