悬浮培养生产口蹄疫病毒灭活工艺研究

<正>采用单独使用二乙烯亚胺(BEI)灭活剂30℃、26℃灭活以及二乙烯亚胺-甲醛(BEI-FA)联合灭活3种方法对悬浮培养生产的猪口蹄疫O型ZK/93病毒液进行灭活,考察和评价各种方法对该毒株146S的影响及灭活安全性。根据对3批口蹄疫病毒灭活后检测结果来看,单独使用BEI26℃灭活方法虽然对146S的损失率小,但存在一定的安全隐患,未经过大量试验证实及确认之前不应使用。而经BEI-FA联合灭活方法获得的抗原安全性检验符合规定,但该方法对     

悬浮培养生产口蹄疫病毒灭活工艺研究

采用单独使用二乙烯亚胺（BEI）灭活剂30℃、26℃灭活以及二乙烯亚胺-甲醛（BEI-FA）联合灭活3种方法对悬浮培养生产的猪口蹄疫O型ZK/93病毒液进行灭活,考察和评价各种方法对该毒株146S的影响及灭活安全性。根据对3批口蹄疫病毒灭活后检测结果来看,单独使用BEI 26℃灭活方法虽然对146S的损失率小,但存在一定的安全隐患,未经过大量试验证实及确认之前不应使用。而经BEI-FA联合灭活方法获得的抗原安全性检验符合规定,但该方法对口蹄疫抗原146S的损失率较高,不宜推广和使用。与上述两种方法相比,用BEI灭活剂30℃灭活28 h仍是最佳的灭活方法。     

四种不同灭活剂对新城疫病毒的灭活效果研究

为探索甲醛、β-丙内酯(BPL)、二乙烯亚胺(BEI)和盐酸聚六亚甲基胍(PHMG)对新城疫病毒(NDV)的最佳灭活条件,试验采用不同浓度梯度的甲醛、BPL、BEI和PHMG对NDV进行不同时间的灭活处理,通过血凝(HA)试验、无菌检验和灭活检验确定灭活效果。结果显示,37℃下,NDV的最佳灭活条件为0.1%～0.2%的甲醛灭活22～24h、0.02%～0.025%的BPL灭活9～10h、2%～3%的PHMG灭活16～20h,该研究为筛选规模化生产ND疫苗所需的理想灭活剂提供了依据。     

犬冠状病毒BEI灭活苗的制备与免疫试验

烷化剂类灭活剂是一类含烷基分子去一个氢原子 ( Cn H2 n 1)化合物 ,它能与另一种化合物作用 ,将烷基引入形成烷基取代物 ,根据其结构可分为氮芥类、乙烯亚胺类和磺酸脂类。这类化合物的化学性质活泼 ,其灭活机制主要在于烷化 DNA分子中的鸟嘌呤或腺嘌呤 ,引起单链断裂、双螺旋键交联 ,妨碍RNA的合成 ,从而抑制细胞分裂。另外 ,也可与微生物的酶系统和核酸蛋白起作用 ,干扰核酸的代谢 ,因此 ,这类灭活剂能破坏病毒的核酸 ,使其完全丧失感染力 ,但并不损害其蛋白质衣壳 ,得以保留其保护性抗原 ,故是制备灭活病毒疫苗较好的灭活剂。据报道…     

二乙烯亚胺对猪细小病毒的灭活作用

使用新型灭活剂二乙烯亚胺(binary ethylenimine,BEI)对猪细小病毒(Porcine parvovirus,PPV)进行了灭活试验,通过ST传代细胞接种法观察病毒灭活后是否出现细胞病变,并结合血凝试验检测灭活效果,确定最佳灭活方法。用3～5日龄乳鼠检测BEI灭活后的病毒培养物和相应制备疫苗的安全性,并用豚鼠检测该灭活工艺制备疫苗的效果,与传统甲醛灭活进行了比较。结果显示,终浓度为1‰的BEI在32℃情况下经20 h即可彻底灭活PPV病毒;BEI灭活的病毒制备的疫苗免疫豚鼠较甲醛灭活病毒产生较高的血凝抑制抗体。     

二乙烯亚胺和甲醛对猫杯状病毒灭活效果比较研究

为比较二乙烯亚胺(BEI)和甲醛对猫杯状病毒(FCV)的灭活条件,采用终浓度分别为0.05%、0.1%、0.2%和0.3%的甲醛和终浓度分别为0.5%、1%、2%和3%的BEI在37℃对FCV灭活6、12、24、36、48 h,通过微量滴定法检测灭活病毒滴度变化,于F81细胞盲传3代验证病毒灭活效果,采用巢式PCR和ELISA法检测病毒抗原完整性。结果显示:37℃下作用48 h,1%、2%、3%的BEI和0.05%、0.1%、0.2%、0.3%甲醛均可灭活FCV。本研究结果可为FCV灭活疫苗的灭活工艺研究提供参考。     

卵黄抗体中外源病毒的不同灭活方法比较

为防止提取的卵黄抗体混入外源病毒,确保提取卵黄抗体的安全性,比较了甲醛和二乙烯亚胺对卵黄抗体的灭活效果。在卵黄抗体中,人工混入鸡新城疫病毒、禽流感病毒（H9型）、禽传染性法氏囊病病毒、减蛋综合征病毒和禽脑脊髓炎病毒,分别加入终浓度为0.02％二乙烯亚胺;0.03％、0.05％和0.08％甲醛, 25℃放置24小时。结果显示,0.02％二乙烯亚胺和0.03％甲醛只能灭活部分病毒,而0.05％和0.08％甲醛能完全灭活上述病毒并且不降低抗体活性。因此,可使用0.05％甲醛灭活卵黄抗体中潜在病毒。     

兽用疫苗的种类及原理

<正>自疫苗注射技术问世以来,兽用疫苗作为一种动物疾病的预防药物,越来越多地应用在畜禽养殖业中。近年来随着生物技术的进步,兽用疫苗的研制获得了快速发展。本文介绍了兽用疫苗的类型和原理。1灭活疫苗该疫苗是把病毒进行灭活后制成的疫苗。选择合适的灭活剂是制备疫苗的关键。目前常用的灭活剂有甲醛、乙烯亚胺和β-丙酸内酯等。2化学合成疫苗该疫苗主要通过化学合成小分子     

产蛋下降综合症(EDS-76)病毒的灭活试验

采用血球凝集抑制试验诊断产蛋下降综合症(EDS—76)时,必须制备一种具有病毒的红血球凝集活性,又不具有感染能力的灭活的病毒作诊断液。作者研究了温度、甲醛溶液和二乙烯亚胺对 DDS—75病毒灭活的影响。     

不同方法灭活卵黄抗体的比较试验

为确保提取卵黄抗体的安全性,防止其中混入外源病毒,为此使用了化学药品灭活的方法。试验对不同终浓度的甲醛和二乙烯亚胺(BEI)对卵黄抗体灭活效果进行了比对。将鸡新城疫病毒(NDV)、禽流感病毒(AIV H9型)、禽传染性囊病病毒(IBDV)、减蛋综合征病毒(EDS-76)和禽脑脊髓炎病毒(AEV)分别混入到卵黄抗体中,混合后加入终浓度为0.03%、0.05%、0.1%甲醛和0.02%BEI于室温条件下,灭活24h,从试验结果我们可以看出0.02%BEI和0.03%甲醛只能灭活部分病毒,而0.05%和0.1%甲醛能灭活全部病毒,并且对抗体活性无任何影响。因此,综合以上结果选用0.05%的甲醛进行灭活,保证疫苗的安全性。     

4株鸡传染性法氏囊病病毒VP2基因序列分析

试验从发病鸡场分离获得4株鸡传染性法氏囊病病毒（infectious bursal disease virus,IBDV）,分别为河北株（HB）、湖北株（HUB）、山东株（SD）、山西株（SX）。通过特异性引物扩增VP2基因并进行测序,序列分析结果显示,4株病毒中,HUB、SD和SX的七肽区为S-W-S-A-S-G-S（aa 326S-332S）,其222（A）、256（I）、294（I）和299（S）位是传染性法氏囊病病毒超强毒株（vvIBDV）的4个特征性氨基酸,确定这3株病毒均为超强毒株;而HB株七肽区为S-W-S-A-R-G-S（aa 326S-332S）,其222（P）、256（V）、294（L）和299（N）位氨基酸显示该毒株属于弱毒株。进化树分析结果显示,HUB、SD和SX 3株病毒与2011年中国分离的大部分IBDV毒株亲缘关系较近,同源性在96%以上,同时与经典毒株Cro-Po/00、超强毒株GX和UK661亲缘关系很近;而HB株与减毒疫苗株D78株和B87株的亲缘关系近。本试验结果表明,IBDV目前在中国流行呈混发型,但以超强毒株为主。     

Germplasm evaluation in beef cattle--Cycle IV: postweaning growth and puberty of heifers.

Postweaning growth, puberty, and pregnancy traits were evaluated for 783 F1 heifers sired by Angus, Hereford, Charolais, Shorthorn, Galloway, Longhorn, Nellore, Piedmontese, and Salers bulls and out of Angus and Hereford dams in Cycle IV of the Germplasm Evaluation (GPE) Program at the U.S. Meat Animal Research Center. The Hereford and Angus sires included a sample of bulls born from 1982 to 1985 (1980s HA) as well as reference sires born from 1963 to 1970 (REF HA) used in previous cycles of the GPE program. Breed group of sire had a significant (P<.01) effect on age and weight at puberty, on 200-, 400-, and 550-d weights, on ADG from 200 to 400 and from 400 to 550 d, and 550-d hip height, but it did not influence (P<.05) pregnancy rate. Mean age and weight at puberty were predicted from the cumulative distribution because of censoring of data in each tail of the distribution. Sire breed group rankings (and predicted means in days) for age at puberty were as follows: Piedmontese (332), Shorthorn (338), Charolais (348), REF HA (348), Galloway (351), 1980s HA (352), Salers (355), Longhorn (357), and Nellore (405). Sire breed group rankings (and predicted means in kilograms) for weight at puberty were Longhorn (283), Piedmontese (298), Galloway (305), REF HA (309), Shorthorn (329), 1980s HA (330), Salers (338), Nellore (341), and Charolais (345). Sire breed group rankings (and least squares means in kilograms) for 200-d weight were Charolais (229), Salers (225), Nellore (221), Shorthorn (220), Piedmontese (215), 1980s HA (215), Galloway (209), REF HA (206), and Longhorn (197), with differences >8.3 kg significant. Rankings for 400-d weight (kilograms) were Charolais (390), Shorthorn (384), Salers (380), 1980s HA (374), Nellore (364), REF HA (356), Piedmontese (353), Galloway (348), and Longhorn (321), with differences >11.5 kg significant. Rankings for 550-d weight (kilograms) were Charolais (445), Salers (430), Shorthorn (429), 80's HA (422), Nellore (420), Piedmontese (401), REF HA (398), Galloway (389), and Longhorn (371), with differences >11.7 kg significant. Rankings for 550-d hip height (centimeters) were Nellore (132.2), Charolais (131.9), Salers (129.9), Shorthorn (129.5), Piedmontese (126.7), 1980s HA (126.1), Longhorn (125.3), Galloway (121.7), and REF HA (121.5), with differences >1.35 cm significant. Breed of sire had significant effects on growth and puberty traits of heifers.     

Feline Hemoperitoneum 16 Cases (1986-1993)

Sixteen cases of feline, non-traumatic hemoperitoneum were evaluated retrospectively. The median age was 5.75 years (range 1.5 - 16 years). There were eight male and eight female cats. Common presenting complaints (n=13) were anorexia (37%), lethargy (31%), and recumbency (31%). Physical examination findings (n=11) included depressed mentation (100%), hypothermia (89%), pale mucous membranes (82%), and poor quality pulses (80%). The median initial peripheral packed cell volume (n=11) was 24% (range 17-55%). In four out of six cases where abdominocentesis was performed, the packed cell volume of the abdominal fluid ranged from 18% to 24%, and matched the peripheral packed cell volume (range 15 - 26%). Some common abnormalities in the serum chemistry screens 9n=6) were elevated alanine aminotransferase in 83% (5/6) of the cats (range 55-5828 U/l) and elevated alkaline phosphatase in 50% (3/6) of the cats (range 18-402 U/l). Ten cats (63%) were euthanized, three (19%) were presented dead on arrival, two (12%) are still alive, and one (6%) were euthanized, three (19%) were presented dead on arrival, two 912%) are still alive, and one (6%) died. The causes of hemoperitoneum were hepatic neoplasia (31%), hepatic necrosis (19%), hepatic amyloidosis (13%), non hepatic neoplasia (13%), hepatopathy (6%), hepatic rupture (6%), necrotic/hemorrhagic cystitis (6%), and ruptured bladder (6%).     

青海可可西里国家自然保护区草地牧草营养成分测定

测定了可可西里国家级自然保护区内紫花针茅草地型和高山嵩草草地型牧草,冷季9月份和4月份营养成分。结果表明,各种牧草营养成分含量随种类不同和生长季节迁移而呈现一定差异;9月份两类草地型牧草DM,CP,EE,NDF,ADF,ADL,Ash,Ca和P平均含量分别为92.20%和90.42%,5.40%和5.98%,2.90%和3.06%,48.30%和44.3%,41.40%和34.36%,3.68%和6.52%,18.85%和8.28%,1.43%和1.25%,0.38%和0.47%;4月份两类草地型牧草DM,CP,EE,NDF,ADF,ADL,Ash,Ca和P平均含量分别为94.33%和93.75%,3.69%和5.85%,1.52%和2.32%,66.82%和61.88%,47.14%和43.32%,9.14%和8.53%,13.48%和13.47%,1.38%和1.16%,0.38%和0.36%;紫花针茅草地型和高山嵩草草地型牧草CP平均含量9月份较4月份分别高出46.8%和35.5%,NDF,ADF和ADL分别降低了27.71%和28.41%,11.95%和20.68%,59.63%和23.33%;就营养品质而言,高山嵩草草地型牧草优于紫花针茅草地型牧草。     

Plasma Cell Myeloma in the Horse

Plasma cell myelomas in horses have been reported infrequently. Data from 10 cases, 9 from the literature and 1 new case, are used to characterize the disease in the horse. Hot-blooded horses (7/10), specifically Quarter Horses (4/10), were most often affected. Median age at diagnosis was 11 years (range, 3 mo-22 yr) and both male (5) and female horses (5) were represented equally. Clinical findings included weight loss (6/8), anorexia (4/8), fever (4/8), limb edema (4/8), pneumonia (3/8), rear leg paresis/ataxia (3/8), epistaxis (3/8), palpable lymphadenopathy (2/8), and bone pain (2/8). Anemia (8/8) was present routinely, and in three horses, RBCs were macrocytic. Leukopenia (2/8), thrombocy-topenia (2/8), and circulating plasma cells (3/8) were variable findings. Except for abnormal protein concentrations and hyponatremia (3), abnormal results from serum biochemical analysis including hypo-cholesterolemia (1), hypercalcemia (1), and azotemia (1) were reported infrequently. Hyperproteinemia (8/9), hypoalbuminemia (7/9), and hyperglobulinemia (8/9) were characteristic but not invariable findings. Monoclonal proteins (7/7) were detected in the α₂, β, or γ region by serum electrophoresis. The paraprotein's heavy chain, determined in four horses, was a subclass of IgG. Three horses had decreased concentrations of normal immunoglobulins. Variable proteinuria (trace to 4+) was detected by routine urinalysis in four of six horses. Bence Jones proteinuria was detected in one of five horses (heat precipitation) and monoclonal proteins were detected in two of three electrophoresed urine samples. Three of the horses had lytic bone lesions detected radiographically. Bone marrow aspirates were diagnostic in two of five horses. Atypical plasma cells or increased numbers of plasma cells or both were present in histologic sections of bone marrow in six of eight horses. Common extraosseous sites of plasma cell infiltration included lymph nodes (8/8), kidney (5/8), spleen (5/8), liver (3/8), lung (3/8), brain (2/8), and orbit (2/8). Two horses had intracellular and extracellular crystalline deposits.     

Occurrence of Escherichia coli O157 in raw material and food in Czech Republic

The study was carried out to investigate the incidence of Escherichia coli O157 in raw materials, foodstuffs and the agricultural environment. Of a total of 987 samples examined, 22 strains (2.2%) were identified as E. coli O157 and 10 of them as E. coli O157:H7. Cefixime-Tellurite MacConkey sorbitol agar (CT-SMAC) agar and Biosynth culture medium (BCM) E. coli O157:7 medium were used for the isolation. The virulence factors (stx1, stx2, eae, and ehxA genes) were identified by polymerase chain reaction (PCR). Most strains were isolated from the mechanically deboned poultry meat (nine), minced meat (six) and raw milk (four). One strain was isolated from beef carcass and two strains from waste water. No strains were were found in mass for sausages, refreshment salads, swabs of pork and poultry carcasses and faeces of cattle and pigs. Ten strains from the 22 identified proved to be positive for all factors of virulence. They were isolated from minced meat (four), raw milk (four), waste water (one) and swab from beef carcass (one). Sensitivity to the antimicrobial drugs ampicillin (AMS), ampicillin-sublactam (SAM), tetracycline (TET), ofloxacine (OFL), cefuroxime (CRX), chloramphenicol (CPM), gentamicine (GEN), colistin (COL), cephalozine (CLZ), cefoxitin (CXT), aztreonam (AZT), and sulphamethoxazole + trimethoprim (COT) was tested using the standard dilution technique and disc diffusion test. Minimum inhibitory concentrations (MIC) characteristics (MIC(50), MIC(90), MIC range) and inhibitory zone diameter were determined for each strain. As determined by MICs, the resistance to tested antibiotics in E. coli O157 isolates was found to AMS (90.9%), CLZ (81.8%), CRX (63.6%), CXT (72.7%), CPM (72.7%), TET (81.8%), SAM (59.1%), COT (9.1%), COL (63.61%), AZT (9%) and GEN (4.5%). The similar results were obtained using the disc diffusion method. The differences were found relating to SAM, CXT, CMO and TET. Resistance against one or more antibiotics was found in 95.4% of E. coli O157. Only one strain was susceptible to all tested antibiotics. Most of the strains were resistant to ampicillin and cephalozine. Eight different resistance phenotypes were demonstrated in E. coli O157.     

Etiological agents of dairy cows' mastitis in western part of Poland

The purpose of the study was to determine the etiological mastitis agents in cows from herds located in the western part of Poland in years 2003-2005. Bacteriological examinations of 18,713 samples taken from California Mastitis Test (CMT) positive quarters were performed according to standard methods. Some Gram-negative bacilli and Gram-positive cocci were examined with the use of API tests (Analytical Profile Index). It was found that 32.7% of samples that were CMT positive were culture negative and 9.3% of samples were contaminated. Streptococcus species (15.7%), coagulase negative staphylococci (CNS) (14.6%), Staph. aureus (8.6%), Gram-negative bacilli (4.0%) and Corynebacterium species (3.8%) were most frequently isolated. Escherichia coli (52.3%) dominated among Gram-negative bacilli followed by Klebsiella pneumoniae (4.1%), Pseudomonas aeruginosa (3.6%), Enterobacter cloacae (3.6%), Serratia marcescens (3.1%), Pasteurella multocida (3.1%), Acinetobacter lwofii (3.1%), and 26 other bacteria species. Stahylococcus xylosus (31.5%), Staph. hyicus (12.2%), Staph. haemolyticus (12.2%), Staph. sciuri (11.6%), Staph. chromogenes (8.8%), Staph. epidermidis (8.3%) and Staph. simulans (6.1%) were the most frequent CNS. Streptococcus uberis (50%), Str. dysgalactiae (19.7%), Str. acidominimus (6.6%) and Enterococcus faecalis (5.3%) were mostly found among CAMP-negative streptococci. An increase in frequency of mastitis caused by non-agalactiae streptococci, Staph. aureus, A. pyogenes and yeast-like fungi and a decrease in that produced by Str. agalactiae in 2005 as compared to years 2003 or 2004 were observed.     

Characterization of multidrug resistant Salmonella recovered from diseased animals

Three hundred and eighty Salmonella isolates recovered from animal diagnostic samples obtained from four state veterinary diagnostic laboratories (AZ, NC, MO, and TN) between 2002 and 2003 were tested for antimicrobial susceptibilities and further characterized for bla(CMY) beta-lactamase genes, class 1 integrons and genetic relatedness using PFGE. Forty-seven serovars were identified, the most common being S. Typhimurium (26%), S. Heidelberg (9%), S, Dublin (8%), S. Newport (8%), S. Derby (7%), and S. Choleraesuis (7%). Three hundred and thirteen (82%) isolates were resistant to at least one antimicrobial, and 265 (70%) to three or more antimicrobials. Resistance was most often observed to tetracycline (78%), followed by streptomycin (73%), sulfamethoxazole (68%), and ampicillin (54%), and to a lesser extent chloramphenicol (37%), kanamycin (37%), amoxicillin-clavulanic acid (20%), and ceftiofur (17%). With regards to animal of origin, swine Salmonella isolates displayed the highest rate of resistance, being resistant to at least one antimicrobial (92%), followed by those recovered from turkey (91%), cattle (77%), chicken (68%), and equine (20%). Serovars commonly showing multidrug resistance (MDR) to > or =9 antimicrobials were S. Uganda (100%), S. Agona (79%), and S. Newport (62%), compared to S. Heidelberg (11%) and S. Typhimurium (7%). Class-1 integrons were detected in 43% of all isolates, and were found to contain aadA, aadB, dhfr, cmlA and sat1 gene cassettes alone or in various combinations. All ceftiofur resistant isolates (n=66) carried the bla(CMY) beta-lactamase gene. A total of 230 PFGE patterns were generated among the 380 isolates tested using XbaI, indicating extensive genetic diversity across recovered Salmonella serovars, however, several MDR clones were repeatedly recovered from different diseased animals.     

广西规模猪场猪呼吸道疾病综合征的病原学调查

为了解猪呼吸道疾病综合征(PRDC)在广西规模场猪群中的感染状况,采用PCR和RT-PCR方法,对2007～2009年期间采自广西13个市65个规模猪场的281份疑似PRDC感染组织样品进行了12种病原体检测。结果显示:73.85%(48/65)的猪场和71.87%(202/281)的组织样品为PRDC感染,其中,被猪繁殖与呼吸综合征病毒(PRRSV)、猪圆环病毒2型(PCV2)、猪瘟病毒(CSFV)、猪链球菌(SS)、猪伪狂犬病毒(PRV)、败血性波氏杆菌(Bb)、猪附红细胞体(E-su is)、猪流感病毒(SIV)、产毒素多杀性巴氏杆菌(T+PM)、副猪嗜血杆菌(HP)、猪细小病毒(PPV)和猪传染性胸膜肺炎放线杆菌(APP)感染(包括混合感染)的阳性样品分别占49.11%(138/281)、34.52%(97/281)、9.60%(27/281)、8.19%(23/281)、6.41%(18/281)、6.41%(18/281)、6.05%(17/281)、6.14%(15/281)、4.63%(13/281)、3.91%(11/281)、0.36%(1/281)和0.00%(0/281);单纯感染占28.11%(79/281);混和感染占43.77%(123/281)。二重、三重和四重混合感染分别占28.11%(79/281)、12.46%(35/281)和3.20%(9/281)。其中,以PRRSV+PCV2(+其他)混合感染形式较多见,占所有混合感染样品的52.03%(64/123)。细菌性病原体感染样品占20.28%(57/281),其中19.22%(54/281)为混合感染。可见:广西地区规模猪场普遍存在PRDC感染,其中PRRSV和PCV2是引起PRDC的主要病原;感染类型复杂多样,以PRRSV+PCV2(+其他)混合感染最为常见。     

奶牛乳房炎金黄色葡萄球菌8种毒力基因的PCR检测

为研究奶牛乳房炎金黄色葡萄球菌(Staphylococcus aureus,SA)主要毒力因子的分布情况,本试验利用PCR对70株临床型乳房炎、55株隐性乳房炎金黄色葡萄球菌的8种主要毒力基因进行检测。结果显示,nuc、ClfA、TSST-1、PVL、Hla、Hlb、FnBPA和FnBPB 8种毒力基因在临床型乳房炎SA菌株中的检出率分别为:100.0%(70株)、100.0%(70株)、0(0株)、5.7%(4株)、100.0%(70株)、11.4%(8株)、97.1%(68株)和100.0%(70株);在隐性乳房炎SA菌株中的检出率分别为:100.0%(55株)、100.0%(55株)、0(0株)、70.9%(39株)、98.2%(54株)、9.1%(5株)、100.0%(55株)和100.0%(55株)。结果表明,nuc、ClfA、Hla、Hlb和FnBPA 5种毒力基因是引起奶牛乳房炎的SA的最主要毒力基因;PVL基因可能是引起隐性乳房炎的重要致病基因。