Bartonella species and their ectoparasites: selective host adaptation or strain selection between the vector and the mammalian host?

A wide range of blood-sucking arthropods have either been confirmed or are suspected as important vectors in Bartonella transmission to mammals, including humans. Overall, it appears that the diversity of Bartonella species DNA identified in ectoparasites is much broader than the species detected in their mammalian hosts, suggesting a mechanism of adaptation of Bartonella species to their host-vector ecosystem. However, these mechanisms leading to the fitness between the vectors and their hosts still need to be investigated.     

1株犬副流感病毒的分离鉴定及生物学特性研究

试验旨在分离鉴定犬副流感病毒(canine parainfluenza virus,CPIV),并对其生物学特性进行研究。用Vero细胞接种感染CPIV阳性犬肺脏组织,盲传4代,收集72 h病毒液进行RT-PCR鉴定、电镜观察、血凝试验、热敏性试验、紫外照射试验及病毒一步生长曲线的测定,同时扩增N基因进行序列分析,并构建系统进化树。结果显示,试验成功从出现咳嗽、流鼻涕等呼吸系统疾病症状的病犬肺脏中分离出1株CPIV,命名为CPIV-BJ01;RT-PCR扩增结果发现,在534 bp处有特异性目的条带。病毒电镜观察发现,其超微结构呈圆形、有囊膜、直径在80～200 nm之间;血凝试验显示,病毒在4和37℃均能凝集1%猪红细胞,与报道的CPIV血凝特性一致;病毒对热敏感,长时间高温下病毒毒价会随之下降;紫外照射可使病毒在短时间内对细胞的感染性急剧下降。病毒一步生长曲线测定结果显示,在12～48 h病毒高速增殖,细胞培养液中病毒滴度急剧上升,之后趋于稳定。CPIV N基因序列与19株有代表性的副流感病毒N基因相比,其核苷酸序列同源性为97.1%～99.8%。遗传进化分析表明,CPIV-BJ01与PIV5 1168-1(登录号:KC237064.1)和PIV5 ZJQ 221(登录号:KX100034.1)位于同一分支上,亲缘关系较近。     

基于Cap蛋白研制的猪圆环病毒2型疫苗研究进展

猪圆环病毒相关疾病(PCVAD)是危害世界养猪业的主要疫病之一,主要由猪圆环病毒2型(PCV-2)引起。该病毒可引起感染猪免疫系统的破坏,造成严重的免疫抑制,继而引发多种病毒/细菌的混合感染与继发感染,给世界养猪业造成了巨大经济损失。近年来疫苗免疫接种是防控PCVAD的有效手段,由于衣壳蛋白(capside,Cap)是PCV-2的主要免疫保护性抗原,因此常被用作研制PCV-2基因工程疫苗的理想靶抗原。论文就PCV-2Cap蛋白基因工程疫苗的研究进展进行综述,以期为今后PCV-2新型疫苗的研究提供参考。     

O型口蹄疫病毒VP1嵌合基因的构建及原核表达

首先合成我国O型口蹄疫病毒2个疫苗毒株VP1基因的3个抗原袁位，与另外扩增的流行毒株VP1基因末端273bp片段相连,构建出O型VP1嵌合基因片段（VP1O）。然后，将VPIO基因连接到原核表达载体pET28a上，构建了重组表达质粒pET28a-VP1O，转化大肠杆菌BL21（DE3）进行诱导表达。SDS-PAGE电泳表明。VP1O基因在大肠杆菌中获得表达。Western blotting检测证实表达的VP1O蛋白具有良好的生物学活性。对表达蛋白通过包涵体洗涤的方法进行初步纯化，获得了较高纯度的VP1O蛋白。     

Calcium homeostasis and its relationship to superoxide production in blood and milk neutrophils of lactating goats

Polymorphonuclear neutrophils (PMN), which comprise over 70% of the somatic cells in goat milk, are a major cellular component of innate immunity in the goat mammary gland. However, the function of milk PMNs is modified after diapedesis compared to PMNs in blood. As many aspects of PMN activity depend directly on intracellular Ca²⁺ concentration ((Ca²⁺)_i), the present study aimed to determine the changes in Ca²⁺ homeostasis of milk PMNs from lactating goats compared to autologous blood PMNs, and to examine the significance of these variations to the immuno-competency of milk PMNs. The intracellular Ca²⁺ store of freshly prepared milk cells was estimated from the elevation of (Ca²⁺)_i after ionomycin treatment, which was found to be significantly less than blood PMNs. Replenishment of the intracellular Ca²⁺ store in milk cells after intracellular Ca²⁺ depletion by Bapta-AM followed by spiking with 2.5 mM Ca²⁺ for 20 min was also compared to that of blood PMNs, showing that after depletion/spiking the intracellular Ca²⁺ store in milk cells was much less than blood PMNs. The production of superoxide anion (O₂^?) in vitro in response to (Ca²⁺)_i-dependent or (Ca²⁺)_i-independent modulators was used to evaluate the relevance of altered Ca²⁺ homeostasis on the immuno-competency of milk cells compared to blood PMNs. The results indicated that milk cells produced similarly low levels of O₂^? as blood PMNs when treated with ionomycin. However, the amount of O₂^? produced by milk cells in response to phorbol 12-myristate 13-acetate (PMA) stimulation, although greater than ionomycin treatment, was significantly less than that of blood PMNs. The capacity for O₂^? production by both cell types in response to PMA reverted to the resting state with use of the protein kinase C (PKC) inhibitor, staurosporine. In conclusion, the current study demonstrated an irreversible shortage of intracellular Ca²⁺ in the milk PMNs of lactating goats compared to blood PMNs. It also showed that preliminary O₂^–production, primed by ionomycin treatment, remained unchanged in milk PMNs, despite the shortage in intracellular Ca²⁺, but decreased O₂^? production capacity, mediated via the PKC pathway, in milk PMN. It is suggested that the defects in Ca²⁺ homeostasis in milk PMNs of lactating goats is partially attributable for the post-diapedesis functionality modifications.     

我国青藏高原地区牦牛草地放牧系统管理及优化

基于牦牛所处地理环境,即青藏高原的特殊性,以及牦牛放牧这一生产方式在该地区第一产业中的重要地位,国内外学者从牦牛放牧系统的管理出发,对该系统中各因子之间的相互响应进行了研究,主要包括当放牧制度不同时,系统中的初级生产力和次级生产力的表现特征。从不同放牧制度中草地和家畜的特征表现,再到人为管理对这两种生产力的影响,研究者通过对比和试验进行了系统优化方法的探索。笔者通过对现阶段已有的研究成果进行整理和分析,概括在不同的放牧管理方式下草地放牧系统中的牦牛在生产力水平方面的差异,以及草地对不同放牧强度的响应,探究青藏高原地区牦牛放牧方式如何影响草畜之间的互作关系。在此基础上,分析其在实际生产活动中存在的问题,以及如何通过改进管理制度优化生产的办法,并对牦牛放牧系统发展进行了展望。     

Effects of road transportation on metabolic and immunological responses in Holstein heifers

This study examined the effects of road transportation on metabolic and immunological responses in dairy heifers. Twenty Holstein heifers in early pregnancy were divided into non‐transported (NT; n = 7) and transported (T; n = 13) groups. Blood was collected before transportation (BT), immediately after transportation for 100 km (T1) and 200 km (T2), and 24 h after transportation (AT). The T heifers had higher (P < 0.05) blood cortisol and non‐esterified fatty acid concentrations after T1 and T2 than did NT heifers. By contrast, the T heifers had lower (P < 0.05) serum triglyceride concentrations after T1 and T2 than had the NT heifers. The serum cortisol and triglyceride concentrations returned (P > 0.05) to the BT concentrations at 24 h AT in the T heifers. The granulocyte‐to‐lymphocyte ratio and the percentage of monocytes were higher (P < 0.05) after T2 in the T heifers than in the NT heifers, suggesting that transportation stress increased the numbers of innate immune cells. T heifers had higher (P < 0.01) plasma haptoglobin concentrations than NT heifers 24 h AT. In conclusion, transportation increased cortisol secretion and was correlated with increased metabolic responses and up‐regulation of peripheral innate immune cells in dairy heifers.     

三聚氰胺对小鼠睾丸病理损伤的观察

本研究采用组织病理学和免疫组织化学技术观察了三聚氰胺（Melamine,MA）对雄性小鼠睾丸的毒性作用,以深入揭示三聚氰胺的毒性作用及其机理。将25只28日龄SPF雄性昆明小鼠随机均分为5组,1组为对照组,其他4组为试验组。4个试验组分别按下述不同剂量灌服三聚氰胺：0.6 mg/kg（bw）/d（每日每公斤体重0.6 mg）、3.0 mg/kg（bw）/d、15.0 mg/kg（bw）/d和17.5 mg/kg（bw）/d,用玉米油做溶剂;空白对照组每日灌服等量玉米油。连续灌胃30天后剖检小鼠并取其睾丸组织进行固定、切片和染色,观察睾丸的组织病理变化。同时,采用免疫组织化学方法检测睾丸组织中增殖细胞核抗原（PCNA）的表达量。观察结果表明,与对照组相比,试验组不同剂量浓度的三聚氰胺对小鼠睾丸组织造成不同程度的病理损伤;三聚氰胺也会在一定程度上影响睾丸组织中PCNA的表达。研究结果表明三聚氰胺对于小鼠生殖系统有明显毒性作用。     

Development of a disperse dye immunochromatographic test for the detection of antibodies against infectious bursal disease virus

For investigating the feasibility of using disperse dyes as an immunoassay chromogenic marker, a disperse dye, DADISPERSE NAVY BLUE SP, was selected in analyzing antibody against infectious bursal disease virus (anti-IBDV). With the color intensity revealed in the disperse dye immunochromatographic test (DICT) strip as the objective function, the optimal dyeing conditions were found as follows: dye concentration absorbance (at lambda(max)=587nm)=3, pH 7, 50 degrees C, for 10min. Under these conditions, the resultant dyed-antibody (rabbit anti-chicken) can produce an optimal color intensity reading of 55,054 on the strip. For performing qualitative immunoassay, chicken sera samples taken from different farms were used for the anti-IBDV titre assessment. The results of DICT strips showed very high sensitivity and specificity as compared to that analyzed by FlockChek enzyme linked immunosorbent assay (F-ELISA) kits. For quantitative immunoassay, it was found that the color intensity measured with DICT was linearly correlated to that of F-ELISA titre (r(2)=0.9687). Therefore, DICT was further applied to the detection of chicken anti-IBDV sera under vaccination in the farms. The average titres of the sampling groups exhibited a strong agreement to that of F-ELISA. Accordingly, the DICT method developed in this study, shown to be reliable, cheap and simple in both qualitative and quantitative immunoassays, is particularly suitable for point-of-need testing (PONT) in agricultural applications.     

西镇牛种质特性研究 三、西镇牛毛色遗传分析

本文从群体遗传学角度对西镇牛的毛色进行了系统分析,确定其毛色遗传由8个位点的等位基因支配,毛色以红黄为主,混有晕毛、有色皮斑、胁及四肢内淡化、黑色背线及黑缟纹等杂色,黑色及白斑在群体中分布频率极低。估测的各位点基因型频率误差小、可靠性高,其毛色分布规律可以做为西镇牛品种特征的遗传标记。同时确认西镇牛毛色形成与中原黄牛和南方黄牛密切相关。