猪圆环病毒2型核酸疫苗的小鼠免疫保护试验

为了研究猪圆环病毒2型(PCV2)核酸疫苗在小鼠攻毒试验中的免疫保护效果,以PCV-2 GXWZ-1株为模板,扩增出ORF2基因及其截短基因8个片段(A(ORF2)、B(51-100aa)、C(101-150aa)、D(181-235aa)、E(151-200aa)、F(51-150aa)、G(101-235aa)、H(51-235aa)),将其插入到pcDNA3.0载体中,构建出真核表达质粒,并将其转染至PK-15细胞,用间接免疫荧光试验检测其瞬时表达情况。将试验小鼠随机分成9组,其中免疫组7组,阴阳性对照各1组,将纯化的真核表达质粒对小鼠进行组合免疫;二免后,用经处理过的PCV-2 GXWZ-2株阳性病料悬液腹腔注射免疫组和非免疫对照组小鼠,阴性对照组用生理盐水腹腔注射;其后进行体重记录、病理切片制作及PCR检测。结果表明:共有6个真核表达质粒成功在PK-15细胞中表达。在攻毒后的3周内,阳性对照组小鼠PCR诊断均为阳性;免疫组中,部分组小鼠在攻毒后第1周或在第2周为阳性,到第3周时各免疫组小鼠全部为阴性;阴性对照组始终为阴性。免疫组在病理保护学方面明显优于非免疫对照组,非免疫对照组的体重增长速率略低于免疫组和阴性对照组。由此可见猪圆环病毒2型核酸疫苗在小鼠攻毒试验中有明显的保护作用。     

小鼠脑提取液中铝的毛细管电泳分析

用毛细管区带电泳法(CZE)测定了小鼠脑中的铝,在10mmol/L苄胺和40％甲醇溶液中,Al3+在7min内与其他组分分离,用间接紫外吸收法在 214nm处检测,并以外标法定量,对样品进行5次测定,RDS为2.9％,回收率为96.8％,方法简便、快速、灵敏度高,重现性好,成本低。     

辣椒种衣剂包衣防疫病机理的研究

研究表明：种衣剂处理对辣椒疫霉菌的侵染过程及辣椒植株的防御酶系均有影响。包衣处理的叶片上休止胞的萌发畸形，不形成附着胞，无力侵入寄主，侵入时间也比对照明显滞后，菌丝在寄主组织内的扩展速度及孢子囊产生的数量明显受到抑制。接种后，过氧化物酶(POD)、多酚氧化酶(PPO)及苯丙氨酸解氨酶(PAL)比对照均有不同程度的提高，出现活性高峰后开始下降。表明甲霜灵对寄主植物抗性的影响具有一定的时间性，且与疫霉菌的激发有关。     

抗丝状支原体丝状亚种单克隆抗体的制备及初步应用

旨在制备抗丝状支原体丝状亚种(Mmm)的特异性单克隆抗体(MAb),为牛传染性胸膜肺炎(CBPP)病原诊断的免疫学方法提供特异性抗体。本研究利用生物信息学技术分析了Mmm国内分离株Ben-1不同传代株的全基因组序列,选取M0071蛋白作为研究对象。将原核表达的可溶性重组蛋白M0071(rM0071)作为免疫原免疫BALB/c小鼠,通过有限稀释法和间接ELISA方法筛选得到能稳定分泌抗rM0071蛋白的单克隆抗体的杂交瘤细胞株。进一步制备单抗腹水并纯化,利用Western blot方法对该单抗进行特异性鉴定,同时测定其抗体效价和抗体亚类。随后利用间接免疫荧光试验(IFA)评价该单抗对细胞感染Mmm的检测能力。结果表明成功获得1株单克隆细胞株3C4A1,将其分泌抗体命名为MAb 3C4A1。特异性结果表明,MAb 3C4A1能与Mmm的分离株和标准株发生特异性反应,而不与山羊支原体山羊肺炎亚种、丝状支原体山羊亚种、牛鼻支原体、无乳支原体、牛支原体、leachii支原体和牛A型巴氏杆菌等发生反应。抗体亚类鉴定MAb 3C4A1属于IgG1亚类、轻链为κ链。经间接ELISA测定其抗体效价为1∶256 000。IFA试验结果表明,MAb 3C4A1仅与感染EBL细胞的Mmm发生绿色荧光反应,而与牛鼻支原体、无乳支原体、牛支原体感染的细胞不发生荧光反应,特异性良好。本研究制备的MAb 3C4A1具有良好的特异性和免疫反应性,可作为CBPP病原免疫学诊断的工具,为进一步研制CBPP病原鉴别诊断试剂盒提供了基础材料。     

羊流产衣原体的分离鉴定及间接免疫荧光法检测

对收集的疑似衣原体感染的羊流产胎儿样本接种鸡胚卵黄囊进行分离培养,通过PCR-RFLP(限制性片段长度多态性分析)方法鉴定为流产衣原体。将收集到的流产衣原体阳性样本接种Hela229细胞,应用基于流产衣原体MOMP单克隆抗体的间接免疫荧光法,在Hela229细胞浆内可见呈绿色荧光的衣原体包涵体,进一步在病原学水平上确定发生在甘肃省肃南县羊流产的病原为流产衣原体。     

The consequence of low mannose-binding lectin plasma concentration in relation to susceptibility to Salmonella Infantis in chickens

Mannose-binding lectin (MBL) is a key protein in innate immunity. MBL binds to carbohydrates on the surface of pathogens, where it initiates complement activation via the lectin-dependent pathway or facilitates opsonophagocytosis. In vitro studies have shown that human MBL is able to bind to Salmonella, but knowledge in relation to chicken MBL and Salmonella is lacking. In order to study this relation day-old chickens from two selected lines L10H and L10L, differing in MBL serum concentration, were either orally infected with S. Infantis (S.123443) or kept as non-infected controls. The differences between healthy L10H and L10L chicken sublines were more profound than differences caused by the S. Infantis infection. The average daily body weight was higher for L10H than for L10L, regardless of infection, indicating beneficial effects of MBL selection on growth. Salmonella was detected in cloacal swabs and the number of Salmonella positive chickens during the experiment was significantly higher in L10L than L10H, indicating that MBL may affect the magnitude of Salmonella colonisation in day-old chickens. MBL expression was determined in ceca tissue by real-time RT-PCR. L10H chickens showed a significantly higher relative expression than L10L at days 1 and 41 pi, regardless of infection. Finally, flow cytometric analysis of whole blood from infected chickens showed that L10H had a significantly higher count of all assessed leucocyte subsets on day 5 pi, and also a higher count of monocytes on day 12 pi than L10L. No difference was observed between infected and non-infected L10L chicken.     

Effects of high‐fat and high‐carbohydrate diets on fat and carbohydrate oxidation and plasma metabolites in healthy cats

High‐fat (HF) or high‐carbohydrate (HC) diets (30% fat, 18.9% carbohydrate; HF and 10% fat, 46.3% carbohydrate; HC) and lengths of adaptation were investigated in cats (Felis catus; 10 ± 2 months, 3.6 ± 0.3 kg). Cats randomly received each treatment for 14 days in a crossover design with a 14‐day washout period between each diet. Three 22‐h indirect calorimetry studies were conducted after acute (day 0), semichronic (day 4) and chronic (day 13) dietary exposure. Blood samples were collected after a 24‐h fast on days 1, 5 and 14. When cats consumed the HC and HF diet, oxidation of the restricted nutrient exceeded intake while oxidation of the nutrient in excess matched intake. Mean max energy expenditure (EE) of cats consuming the HF and HC diet were 107 and 102 kcal/kg^0.67/day and occurred at a mean of 4 and 12 h post‐feeding respectively. Maximal fat (0.90 g/h) and carbohydrate (carbohydrate; 1.42 g/h) oxidation were attained at 26 min and 10.4 h post‐feeding respectively. The changes observed in macronutrient oxidation and EE suggest that cats adapt whole‐body nutrient metabolism in response to changes in dietary macronutrient content, but may require longer than 14 day to adapt to a macronutrient that is present at a lower concentration in the diet.     

草鱼巨噬细胞的集结及相关的组织变化

给草鱼腹腔注射细微炭粒（墨汁）后不同时间取样,比较观察了草鱼胸腺、肝、脾、头肾和肾组织中巨噬细胞集结（ｍ ａｃｒｏｐｈａｇｅ ａｇｇｒｅｇａｔｉｏｎ , Ｍ Ａ）状况及相应组织的变化。结果表明,在头肾和脾组织中, Ｍ Ａ 的数量和体积均较肝和肾中的多而大,胸腺中所产生的 Ｍ Ａ 数量较肝和肾中的少,体积也小;注射墨汁后 １～３ 周内,在胸腺、头肾、肝和脾组织中, Ｍ Ａ 的数量和体积均有不同程度的增加,但在注射墨汁后第 ４ 周的肾组织中, Ｍ Ａ 的体积缩小而且密度降低,同时肾组织结构有逐渐恢复的趋势,可能是由于 Ｍ Ａ 分解后通过肾小球和肾小管排泄到膀胱或体外的缘故。对草鱼 Ｍ Ａ 的组织分布、排除等分析认为, Ｍ Ａ 可作为草鱼非特异细胞防御作用的解剖学和细胞学的一种生物标记。