鱼类抗弹状病毒感染的免疫机制

弹状病毒属于单链RNA病毒,是威胁野生和养殖鱼类的重要致病因子.简要介绍了鱼类应对弹状病毒感染的天然性免疫和适应性免疫机制,以及病毒逃避免疫防御策略,以阐明鱼类与弹状病毒相互作用机制,为开发防御病毒感染的新疫苗和治疗措施提供参考.     

非洲猪瘟病毒p54蛋白单克隆抗体制备及其抗原表位鉴定

旨在制备非洲猪瘟病毒(ASFV)p54蛋白的特异性单克隆抗体。本研究利用大肠杆菌表达系统表达p54蛋白,免疫BALB/c小鼠,取其脾细胞与SP2/0细胞进行细胞融合。利用纯化的p54蛋白作为包被抗原,采用间接ELISA方法筛选获得阳性杂交瘤细胞。经4次亚克隆后,取杂交瘤细胞上清进行单克隆抗体亚型鉴定,利用体内诱生法制备单克隆抗体并进行纯化。间接ELISA方法检测单克隆抗体的效价,利用交叉反应性试验、间接免疫荧光试验和蛋白印迹对所获单克隆抗体的特异性进行鉴定。根据预测的p54蛋白二级结构,采用逐步截短法分析鉴定单克隆抗体识别的抗原表位区域,并在p54的三级结构中进行标注。结果显示：成功筛选了6株分泌p54单克隆抗体的杂交瘤细胞,分别命名为28G12-1、31G7-1、31G7-2、35F10-1、35F10-2、38D3-1。其中28G12-1、31G7-1、31G7-2重链为IgG2a型,35F10-1、35F10-2、38D3-1重链为IgG1型;轻链均为κ链。单克隆抗体的最低效价为1∶25 600,与猪繁殖与呼吸综合征病毒、猪伪狂犬病病毒、猪流行性腹泻病毒、猪细小病毒、猪急性腹泻综...     

长毛兔的行为观察

本文对长毛兔的行为的系统观察所得的结果作了较详细的记述。长毛兔的夜间活动多于白天,因此,无论其采食量和饮水量以及排粪量和排尿量等夜间均高于白天;并呈现明显的季节性变化。长毛兔的各种行为均受环境条件的影响,不良的环境会扰乱长毛兔的生理活动和行为表现,导致生产性能下降。通过行为观察,对长毛兔的饲养管理方面与其行为或生理变化的相互关系问题,在理论上和实际生产上都获得有益的启示和知识,对于制定高效率的生产措施具有重要意义。     

蜜蜂螺原体单克隆抗体的制备及其基本性状研究

用蜜蜂螺原体(CH-1)免疫的 BALB/C 小鼠脾细胞与SP2/0鼠骨髓瘤细胞融合,获得3株稳定分泌 CH-1单克隆抗体(简称单抗,下同)的杂交瘤细胞株。腹水单抗效价介于12800~(-1)～6553600~(-1)之间。其免疫球蛋白种类为 IgG1和IgG3。腹水单抗与同源抗原的反应能被兔多克隆抗血清阻断。杂交瘤细胞染色体为96条。反复冻融、低温保存以及 pH 试验对单抗有一定影响。单抗与中国的蜜蜂螺原体分离物有反应,而与美国蜜蜂分离物无反应。     

Two monoclonal antibodies for the recognition of Mytilus spp. larvae: studies on cultured larvae and tests on plankton samples

Monoclonal antibodies (mAbs) were generated against 2-day-old mussel larvae in an attempt to develop a rapid and rigorous method for the identification of mussel larvae in field plankton samples. Previously, we have shown that two of these mAbs recognised Galician Mytilus galloprovincialis obtained from monospecific cultures, but did not recognise the larvae of other bivalve species present in that area. To assess the possibility of using these mAbs in routine assays for measuring the abundance of mussel larvae in plankton, studies on cultured mussel larvae, at different stages of development, and tests on bivalve larvae from plankton samples were carried out. Initially, to see whether the two mAbs also recognise other mussel larval stages, they were tested against mussel larvae of different ages obtained from monospecific cultures. The results indicate that both antibodies stain all the stages tested, even 1-month-old postlarvae. In addition, we also demonstrate that these mAbs also recognise other forms of Mytilus. Both antibodies bind to M. galloprovincialis larvae from the Mediterranean Sea and M. edulis larvae. Finally, and more significantly, studies on field plankton samples were performed to confirm if both mAbs are really mussel-specific, and do not cross-react with larvae of any other bivalve species existing in the plankton. The results presented here clearly indicate that our two monoclonal antibodies specifically recognise the mussel larvae in field plankton samples from different geographical regions, but not the larvae of any other bivalve species. Thus, these monoclonal antibodies could be used for routine monitoring of mussel larvae in plankton samples from different sources.     

The effect of anti-Paramoeba antibodies on Paramoeba sp., the causative agent of amoebic gill disease

The effect of anti- Paramoeba antibodies produced in sheep on Paramoeba sp., the causative agent of amoebic gill disease (AGD), was tested. Incubation of Paramoeba sp. with antiserum at a concentration of 20% or more effectively coated the Paramoeba with antibodies. The antiserum had no effect on Paramoeba sp. survival in vitro or on its ability to cause AGD in Atlantic salmon.     

Evaluation of a West Nile virus surveillance and early warning system in Greece,based on domestic pigeons

In the summer of 2010 an epidemic of West Nile virus (WNV) occurred in Central Macedonia, Greece, with 197 human neuroinvasive disease (WNND) cases. In the following years the virus spread to new areas, with a total of 76 WNND cases in 2011, and 109 WNND cases in 2012 (14 and 12 WNND cases, respectively, in Central Macedonia). We established a surveillance system based on serological testing of domestic pigeons, using cELISA confirmed by serum neutralization test. In Central Macedonia, pigeon seroprevalence was 54% (95% CI: 49–59%) and 31% (95% CI: 24–37%) at the end of the 2010 and 2011 epidemic seasons, respectively. One serum was positive for neutralizing antibodies directed against Usutu virus. Pigeon WNV seroprevalence and incidence rates of human WNND after the 2010 epidemic were positively correlated (ρ = 0.94, at the regional unit level), while in 2011 the correlation (ρ = 0.56) was not statistically significant, possibly due to small number of human WNND cases recorded. To evaluate the efficacy of the system at alerting upon WNV enzootic circulation before the onset of human cases, we tested 270 pigeons in 2011 and 240 pigeons in 2012. In Central Macedonia, the first seroconversions in pigeons were recorded 44 and 47 days, respectively, before the first human WNND cases. Pigeon surveillance was used successfully for identification of areas with WNV enzootic transmission and for early warning. Timely diffusion of information to health authorities facilitated the implementation of preparedness plans to protect public health.