猪繁殖与呼吸综合征病毒ORF7基因的表达和重组蛋白的纯化及免疫学活性分析

[目的]在基因工程菌中实现猪繁殖与呼吸综合征病毒（PRRSV）ORF7基因的高效表达,对表达的重组蛋白进行纯化,并探讨其免疫学活性及应用价值。[方法]将已构建好的重组表达载体pET-ORF7转化大肠杆菌BL21（DE3）,用IPTG在最适条件下诱导表达,其表达产物用SDS-PAGE和Western Blot进行检测。应用Ni-NTAHis.Bind Resin层析柱在变性条件下纯化表达产物,通过梯度透析进行复性。采用Western Blot及间接ELISA法检测复性后重组蛋白的免疫学活性。[结果]重组质粒pET-ORF7在大肠杆菌中以融合形式成功表达,融合表达产物主要以包涵体形式存在。SDS-PAGE检测所表达的重组蛋白分子量为33 kD左右,与预期大小相符。Band-scan软件对表达蛋白分析发现,表达产物约占菌体蛋白的50%。Western Blot及间接ELISA法显示复性后的重组蛋白与PRRSV阳性血清有特异性结合反应,表明其具有良好的免疫学活性。[结论]该试验将为进一步研制PRRSV单克隆抗体及诊断试剂盒奠定基础。     

规模化猪场猪繁殖与呼吸综合征感染混合细菌研究

规模化猪场爆发急性高热病,采集死猪肺脏病料和血清5份用RT-PCR方法进行PRRSVNSP2基因扩增,3份为猪繁殖与呼吸综合征阳性,编号为scwhn01,scwhn02和scwhn03.对scwhn01 NSP2基因进行克隆和序列测定分析,有30个氨基酸的不连续缺失,与VR2332,JXA1,BJ-4,HuN4,CH-1a,HB-1(sh)2002和HB-2(sh)2002的NSP2的同源率在74.2%~91.0%之间,并从病料中分离到2株细菌,编号分别为SNC0901和SNC0902,均使被攻毒小鼠死亡.16S rDNA测序并在Genbank中blast鉴定为副猪嗜血杆菌和大肠杆菌.对所分离的细菌进行17种抗生素药敏实验,对头孢噻呋、头孢噻吩等敏感;对克林霉素、阿莫西林/棒酸等中度敏感;对荷兰、林可霉素、丁胺卡那霉素和土霉素等耐药.表明该猪场因高致病性蓝耳病病毒混合感染副猪嗜血杆菌、大肠杆菌导致猪大量死亡.     

不同施氮处理对水稻颖果灌浆和呼吸活性的影响

为了研究施氮量和施氮时期对水稻颖果生理活性的影响,以一籼一粳两个水稻品种为材料,施用分蘖肥和孕穗肥,并在施氮总量相同的条件下设置了前高后低和前低后高两个处理。研究结果表明：增施氮肥维持了水稻颖果灌浆后期较高的含水率,延长了颖果灌浆充实的时间;维持了水稻颖果灌浆充实期尤其是灌浆后期较高的呼吸速率,并且使颖果维持较高呼吸活性的时间延长;延长了颖果各组织维持脱氢酶活性的时间。其中孕穗肥较高的施氮处理比分蘖肥较高的施氮处理效果更为显著。     

Study on the glass transition for several processed fish muscles and its protein fractions using differential scanning calorimetry

ABSTRACT:   The glass transition behavior of processed fish muscles (bonito, tuna, mackerel, sea bream, cod) and its muscle protein fractions (sarcoplasmic and myofibrillar proteins) were studied using differential scanning calorimetry. Each dried processed fish muscle and the extracted protein fractions showed clear glass transition phenomenon. The T _g values of muscles and myofibrillar proteins from red muscle fishes tended to be lower than those from white muscle fishes though there was no difference on T _g of sarcoplasmic proteins. The T _g value of whole muscle was considerably lower than that of extracted protein fractions because of the plasticizing effects of low molecular weight materials contained in the muscle.     

外源ABA对冬小麦越冬期呼吸代谢关键酶与糖代谢的影响

为探讨外源ABA对低温胁迫下冬小麦幼苗呼吸代谢的影响,以抗寒品种东农冬麦1号和冷敏感品种济麦22为材料,测定和分析了小麦三叶期叶面喷施ABA(浓度10-5 mol·L-1,剂量为每4 m2喷施1 L)后越冬期叶片和分蘖节在低温胁迫下碳水化合物含量、糖酵解代谢酶及三羧酸循环代谢酶活性的变化.结果表明,越冬期间随温度的降低,两个小麦品种的叶片和分蘖节中可溶性总糖和淀粉含量先升后降;参与糖酵解代谢的丙酮酸激酶、己糖激酶、磷酸果糖激酶及三羧酸循环代谢酶的活性在对照叶片中均先升后降,但在分蘖节中的变化则不尽相同.外源ABA处理使低温下两个小麦品种叶片和分蘖节积累更多的可溶性糖和淀粉,并且不同程度地提高了两个品种叶片及分蘖节中丙酮酸激酶、己糖激酶及磷酸果糖激酶的活性,尤其在最低温-25℃时明显抑制这几种酶活性的降低,这种影响在抗寒性强的东农冬麦1号中更明显;在低温胁迫后期抑制了三羧酸循环代谢酶和丙酮酸脱氢酶活性的降低,使其保持在相对较高的水平.外源ABA对碳水化合物积累的促进作用有利于植株呼吸和ATP合成维持在较高水平,进而提高低温下植株存活能力.     

双壳贝类呼吸代谢的研究进展

呼吸代谢是生物能量学研究的重要内容之一。依据近年来国内外的研究成果,综述了关于贝类呼吸代谢的研究方法,讨论了影响贝类耗氧率的生物因素和非生物因素,展望了贝类呼吸代谢的研究和应用前景。     

人呼吸道合胞病毒的研究与防治进展

人呼吸道合胞病毒是全世界婴幼儿下呼吸道感染最常见的病原体,导致每年超过300万人住院和20万以上的人死亡,给全球带来了非常严重的经济和社会负担。目前,尚不存在防治RSV的疫苗,也没有特异性强、安全性高的抗RSV药物。对HRSV的结构、流行病学特征以及疾病模型构建方面的研究进展进行综述,并对已有的检测和治疗方法进行总结,旨在为今后开发抗HRSV的药物提供全面的参考信息。     

Clinical efficacy of diclofenac sodium and flunixin meglumine as adjuncts to antibacterial treatment of respiratory disease of calves

Objective To compare the efficacy of the non-steroidal antiinflammatory drugs, diclofenac sodium and flunixin meglumine as adjuncts to the antibiotic treatment of bovine respiratory disease (BRD). Procedure We randomly allocated 80 Holstein calves with BRD to three groups. All the calves received a dose of 2.5 mg/kg tulathromycin by single subcutaneous injection and two of the groups received, in addition, either 2.5 mg/kg diclofenac sodium as a single intramuscular injection (diclofenac group, n = 30) or 2.2 mg/kg flunixin meglumine as an intravenous injection on the first three consecutive days after tulathromycin administration (flunixin group, n = 30). All calves were given a clinical score prior to initial treatment (day 0) and after treatment (days 1, 2, 3, 7 and 14) by observing appetite, demeanour, rectal temperature, the rate and type of respiration, presence or absence of coughing, and nasal discharge. Results During the first 48 h, improvement of adverse signs of respiratory disease, such as pyrexia and elevated respiratory rate, and of a high clinical index score was significant in the two adjunct groups compared with the calves receiving antibiotic alone. The reduction in pyrexia was greatest in the diclofenac group. There were no statically significant differences between treatment groups with regard to eventual perceived recovery from respiratory disease in 14 days. Conclusion In this trial, a single intramuscular dose of diclofenac sodium was equally effective as three intravenous injections of flunixin meglumine given on consecutive days as adjunctive therapy for BRD.     

An Outbreak of Respiratory Disease Complex in Sheep in Central Ethiopia

An investigation was carried out into an outbreak of respiratory disease complex (RDC) in 3641 Menz and Awassi×Menz cross sheep in Central Ethiopia between 1998 and 1999 by clinical, serological, microbiological, post-mortem and histopathological examinations. The monthly incidence of RDC varied from 2.8% to 4.0% and the prevalence was as high as 17%. The case fatality rate was 18%, despite culling of sick sheep. Over 76% of the morbidity occurred in adults, followed by 19% among weaners. Similarly, 62% of the mortality was in adults. However, 27% of the mortality occurred in lambs despite the low morbidity in the group. Significant breed and age differences were seen in the morbidity and mortality rates (p<0.05). Clinical signs, gross and microscopic lesions and serological and bacteriological examinations showed an interplay of several causes of the RDC, including pestes des petits ruminants (PPR) (72.3%, serologically confirmed), lung worms, maedi-visna, bacterial bronchopneumonia (staphylococcal and streptococcal), enzootic pneumonia and some fungal infections. Cold temperatures, which may be as low as –8.5°C at night, are major predisposing factors along with managemental stresses. Vaccination of animals with a homologous PPR vaccine appeared to decrease dramatically the occurrence of the disease, showing that PPR played an important role in the outbreak. Several of the pathogens do not appear to be individually capable of causing the respiratory disease. Appropriate strategies for the prevention of RDC are suggested.