非洲猪瘟病毒无标签重组B602L抗原制备与鉴定

为了获得无标签重组抗原用于非洲猪瘟病毒(ASFV)抗体检测,本研究将B602L与类弹性蛋白多肽(ELP)基因进行融合表达,利用简单、经济的相变循环(ITC)纯化ELP-B602L融合蛋白,用烟草蚀纹病毒(TEV)蛋白酶活性包涵体切除ELP标签,再用ITC回收重组B602L蛋白;用抗体阳性猪血清对重组B602L进行鉴定;以重组B602L蛋白为包被抗原进行ELISA鉴定。结果显示重组大肠杆菌能正确表达ELP-B602L融合蛋白,纯化融合蛋白纯度大于85%;TEV蛋白酶活性包涵体切除ELP标签的效率大于90%,回收的重组B602L蛋白纯度大于90%,能与抗体阳性猪血清反应;以重组B602L蛋白为包被抗原建立的ELISA与抗体阳性血清反应为阳性,与抗体阴性血清反应为阴性,OD450值与血清稀释倍数具有线性相关性。     

屠宰环节检疫检验印章印油使用现状及问题分析

本文调研并整理了屠宰环节现行有效的检疫检验印章种类及印油的使用情况,分析了实际应用中存在的问题和潜在风险,并就进一步规范屠宰环节检疫检验印章及印油的使用和管理提出了建议。     

Dynamics of natural bovine herpesvirus-1 (BoHV-1) infection in a dairy herd

In this study, natural cycling of BoHV-1 infection was investigated in two groups of dairy cattle containing 2120 head. Group 1 comprised 127 animals and they were monitored for BoHV-1 infection virologically and serologically in six consecutive sampling periods. It consisted of naive heifers between 6 and 8 months of age, while in group 2, age, sex and the BoHV-1 serostatus of the animals were disregarded. The animals in group 1 were found to have seroconverted at the second sampling. Results of the serological study showed slight antibody response after natural BoHV-1 infection in the herd and neutralizing titres fell below protective levels in the 6–8 months after the peak. During the 2-year study period, one recurrence was detected after primary infection. Virus isolation studies revealed a cytopathic effect indicative of BoHV-1 in two nasal swabs taken during the fifth sampling period from animals with mild upper respiratory tract symptoms. As the study was carried out under natural conditions, it is not known whether the viruses isolated were from recurrences or re-infections. Data from cross-neutralization tests with herd isolates showed higher antibody response than those with the reference virus. The dynamics of BoHV-1 in both groups were found to be statistically similar.     

检测流感病毒的商业PCR试剂盒比对验证

利用8种不同流感病毒株及其核酸,结合自然感染组织样品与人工混合样,验证比较了8种商业PCR试剂盒的质量参数。结果表明,3号荧光PCR试剂盒漏检1株外,其他试剂盒均可以检测8种毒株,对其他类似病毒如猪繁殖与呼吸综合征病毒、猪伪狂犬病毒、猪传染性胃肠炎病毒、猪细小病毒和猪环状病毒核酸等无交叉反应,最高可检测到10-7稀释的目标核酸,重复性与再现性均为理想,在有效期内检测结果没有发生变化。     

Immunoadjuvant effects of hemagglutinating virus of Japan envelope (HVJ-E) on the inactivated H9 subtype avian influenza virus vaccine

Avian influenza viruses (AIV) of the H9 subtype cause serious health problems in chickens, resulting in great economic losses to the poultry industry worldwide. The killed vaccine (KV) against H9 subtype AIV has been widely used in China since 1998 but has been linked with side effects in chickens and only partial protection. A few studies have demonstrated the immunostimulatory effects of the hemagglutinating virus of Japan envelope (HVJ-E) in cancer therapy. In this study, the adjuvant efficacy and the protective effects of HVJ-E, in combination with H9N2 AI KV against AIV were evaluated. The maturation of murine dendritic cells treated by HVJ-E was verified by FACS in the current experiment, then the antibody hemagglutination inhibition (HI) titers and cytokines and the post-challenge virological profiles (oropharyngeal and cloacal virus shedding) were investigated to define the immune responses in chickens. Our findings indicate that HVJ-E could induce dendritic cell (DC) maturation in mice. Injection of HVJ-E in chickens resulted in raised levels of IFN-β and IFN-γ being present in sera suggesting a stimulatory effect in these animals. The antibody responses to AIV of chickens inoculated with HVJ-E adjuvanted killed H9-AIV were higher than those of chickens inoculated with oil adjuvanted H9-HIV. Furthermore, although inoculation of either HVJ-E or oil adjuvanted AIV reduced virus shedding following challenge, compared to controls, HVJ-E adjuvanted AIV was more effective in reducing shedding than oil adjuvant.     

新牧1号苜蓿两个抗逆相关基因启动子的克隆及分析

采用实时荧光定量PCR分析了新牧1号苜蓿（Medicago varia Xinmu 1）MvNHX1和MvDREB1基因在盐胁迫下的表达情况。此外,根据已获得的MvDREB1和MvNHX1基因序列设计特异引物,并以新牧1号苜蓿的基因组DNA为模板,克隆得到了这两个基因的启动子。利用生物信息学方法,分析这两个基因启动子的类型和结构,结果表明,MvNHX1和MvDREB1基因的启动子序列中均含有通用启动元件和上游调控元件,如CAAT框、TATA框、光响应元件、低温响应元件等,但部分响应元件的种类和数量不同。通过对两个基因启动子的克隆、分析及比较为进一步研究这两个基因的表达调控机制奠定了基础。     

基于猪丁型冠状病毒重组M蛋白间接ELISA抗体检测方法的建立与应用

猪丁型冠状病毒(porcine deltacoronavirus,PDCoV)是近年新发现的一种猪肠道病原体,主要感染仔猪并引起仔猪腹泻甚至死亡,针对该病建立快速、准确的血清学检测方法尤为重要.本研究以PDCoVHNZK-04毒株为模板扩增M基因并将其克隆至原核表达载体pET-32a中,构建了重组质粒pET-32a-P...     

Frequent use of colistin-based drug treatment to eliminate extended-spectrum beta-lactamase-producing <Emphasis Type="Italic">Escherichia coli</Emphasis> in backyard chicken farms in Thai Binh Province,Vietnam

Reports of livestock infections with extended-spectrum beta-lactamase-producing Escherichia coli (ESBL-E) are increasing. Based on interviews conducted over a 6-month period, we found that veterinarians in the Vietnamese province of Thai Binh prefer to prescribe colistin-based drugs (CBD) in chicken farms. We aimed to clarify whether CBD use selects for strains of colistin-resistant ESBL-E. With the cooperation of seven local households, we detected ESBL-E in chickens’ feces after treating chickens with CBD. Phylogenetic groupings and the presence of CTX-M/AmpC genes were determined, and the multi-antibiotic susceptibility of isolates was analyzed. Our results showed that ESBL-E presented in seven chickens’ feces from two households. Seventy-two percent of ESBL-E isolates harbored CTX-M9 and the phylogenetic group A; the colistin minimum inhibitory concentration (MIC) of all isolated ESBL-E ranged from 0.064 to 1 μg mL^?1. Moreover, ESBL-E isolates were used to experimentally select for colistin resistance, and the effect of commercial CBD on ESBL-E was investigated. The results showed that an ESBL-E strain with a colistin MIC of 4 μg mL^?1 was able to grow in media with CBD. Although CBD treatment was effective, in vitro experiments demonstrated that ESBL-E can easily acquire colistin resistance. Therefore, restrictions on colistin use are necessary to prevent the emergence of colistin-resistant bacteria.     

Effect of herbicides with different modes of action on physiological and cellular traits of Anabaena fertilissima

Cyanobacteria are important components of the lowland rice ecosystem. Therefore, it is important to examine the effect of herbicides (commonly used against weeds of rice crop) on the performance of cyanobacteria. We studied the toxic effects of three herbicides often used in rice field, viz. propanil, pretilachlor and glyphosate, on the performance traits of Anabaena fertilissima C.B. Rao. Pretilachlor [0–40 active ingredient (ai) mg/L] and glyphosate (0–80 ai mg/L) exhibited toxicity to A. fertilissima at higher doses than propanil (0–1.5 mg/L). Propanil had severe damaging effects on cellular characteristics of A. fertilissima when compared to pretilachlor or glyphosate. Propanil treatment of A. fertilissima resulted in the leakage of protoplast from the heterocyst due to the breakage of the plasma membrane and surrounding wall. Our study shows that photosystem II herbicides such as propanil could have deleterious effects on phototrophic (cyanobacterial) communities, which are an integral part of the rice ecosystem.     

Ethnoveterinary health management practices using medicinal plants in South Asia – a review

Animal rearing is the major occupation of most population of South Asian countries. Due to lack of resources and limited approach to modern medicine, most of the livestock raisers prefer to use plant-based traditional medicine also referred to as ethnoveterinary medicine (EVM). Indeed, the use of medicinal plants in South Asia dates back to several centuries with documented evidences. However, there is currently a dearth of documentation and compilation of use of medicinal plants for animal diseases in this part of the world. This review aims to provide an up-to-date compilation of common medicinal plants used for the treatment and/or management of common animal diseases in South Asian countries. Extensive literature search was conducted online and relevant data was retrieved from well-known scientific databases. A total of 276 plants belonging to 95 families have been documented to be in common use for managing 14 different categories of animal diseases. Solanaceae, Lamiaceae, Fabaceae, and Leguminosae were most common plant families in terms of their plant species used for EVM. Gastric diseases were commonly reported and accounted for 72 species of plants used for its treatment followed by the miscellaneous disorders category and skin diseases comprising of 65 and 39 plant species respectively. Herbs accounted for 46% of the total plant species, followed by trees (33%), and shrubs (18%). The EVM were applied through different routes of administration; oral administration accounted for 72% followed by topical application 27%, while burning of plant parts to create smoke around animals to repel insects was less common (1%). It is anticipated that the present review will stimulate further ethnoveterinary research among livestock disease management practices in South Asia.