一种轮状病毒特异抗体ELISA检测方法

在实验室条件下,建立较为快速的轮状病毒抗体ELISA检测方法。利用组织培养的轮状病毒SA11株经初步的浓缩和纯化后,作为包被抗原包被酶标板。SDS-PAGE分析表明,经浓缩纯化后,轮状病毒的主要抗原均存在。将测试血清用SA11中和后,轮状病毒或其抗原免疫血清(肠黏液,粪便悬液)的OD490值比中和前有明显的下降,下降比率大于45％,而对照组血清的OD490值与中和前相比没有明显的变化,下降比率小于9％。表明用此ELISA体系检测轮状病毒抗体具有特异性。     

长沙生态动物园野生动物消化道寄生虫感染情况调查

为了解长沙生态动物园动物感染的寄生虫种类及感染情况,采用饱和盐水漂浮法和沉淀法检查了57种动物的粪样。结果表明:长沙生态动物园动物寄生虫感染率为52.63%,其中食草类动物和灵长类动物阳性率为50%,食肉类动物感染率最高,感染率达76.9%。动物园动物常见感染的寄生虫有球虫、吸虫、线虫、绦虫等,其中线虫感染率最高。     

动物园动物体内寄生虫的调查

为了摸清我市龙沙公园内展出的野生观赏动物寄生虫的感染情况 ,1991年以来随机对齐齐哈尔市龙沙公园的 33种 2 2 5头 (只 )的不同动物进行了以肠道寄生虫为主的 (不包括皮肤寄生虫 )调查 ,基本查清了感染情况 ,对临床治疗、动物管理和人员防护将起到协助作用。报告如下。1　普查方法1.1　蠕虫学常规粪检法收集虫卵。饱和食盐浮集法、反复洗涤沉淀法等 ,对肠道寄生原虫除直接涂片碘着色观察外 ,并用铁苏木素染色法 ,制成标本保存 ,以便鉴定虫体。1.2　用蠕虫学剖检对尸体进行虫体收集。1.3　对临床检查寄生虫阳性的动物进行驱虫 ,对驱出的虫…     

上海动物园游客保护教育需求及满意度调查

本研究利用问卷调查收集了上海动物园游客类型与特征、游园目的、游客游览时获得野生动物科普知识的途径、游客对现有科普活动的评价、对科普教育的需求、游客满意度、意见与建议等,并进行了研究。结果表明,大部分(66.02%)游客为本市居民和暂居本市的中青年。近9成(89.51%)游客携带孩子游览动物园,其中超过半数(52.34%)为父母与孩子共同出行。游客游园目的主要为观看动物、游玩娱乐和让孩子学习动物知识。观察动物和阅读动物科普说明牌是游客学习动物科普知识的主要途径。互动类和深度游类项目,是游客最愿意参加的活动类型。游客更希望园方从动物科普设施、饲养员讲解、志愿者讲解这3方面提升其教育功能。总体而言,游客对上海动物园感到满意,超过半数(53.81%)表示很满意。具体而言,动物说明牌的满意度最高,其次是路面状况和路标指示牌。基于研究结果,提出以下建议:提升科普活动的内涵和互动性;提升游园体验,延长游览时间;寻找迎合和引导的平衡;完善基础设施。     

A群猪轮状病毒VP7蛋白单克隆抗体的制备及鉴定

To prepare the monoclonal antibodies against VP7 protein of group A porcine rotavirus (PoRV) serotype G11,VP7 gene of PoRV serotype G11 was cloned into the vector pGEX-6P-1. The recombinant plasmid was transformed into E.coli competent cells DH5α and induced by IPTG. Hybridomas were produced by fusing SP2/0 cells with spleen cells from mouse immunized with GST-VP7 recombinant protein. One hybridomas secreting MAb against VP7 protein was identified by indirect ELISA. Western blotting analysis showed that the MAb could recognize the recombinant VP7 protein and authentic VP7 protein of PoRV，and the specific immunoflurescence was detected in PoRV infected MA104 cells by indirect immunoflurescence assay.The result of MTT method showed that the MAb had the neutralizing activity. The subtype of the MAb was IgG2b. The results showed that VP7 protein was successfully expressed in E.coli， one MAb was preparated and identified，and it could be used for further study of prevention，diagnosis and treatment of porcine rotavirus disease.     

动物园部分动物舍内外环境中常见病原菌的调查

为了解动物园动物及其环境中常见病原菌的分布情况,采用菌落计数、细菌分离培养及动物致病性试验等方法,对南京市红山森林动物园的部分动物生存环境的空气、土壤、水共126份样品进行检测.结果表明,各种动物舍内菌落总数均大于舍外菌落总数.从126份样品中分离到16个菌种127个菌株,其中葡萄球菌有69株,占54.33%(69/127);肠杆菌有51株,占40.16%(51/127);链球菌有7株,占5.51%(7/127).通过动物试验得知,分离得到的金黄色葡萄球菌、大肠杆菌、臭鼻克雷伯氏菌、粘质沙雷氏菌、假结核耶氏菌、福氏志贺氏菌1-5型、聚团肠杆菌和血链球菌等8种致病性较强,松鼠葡萄球菌、表皮葡萄球菌、解糖葡萄球菌和变异链球菌等4种致病性较弱,耳葡萄球菌、头葡萄球菌、施莱福葡萄球菌和唾液链球菌等无致病性.动物舍内外环境中葡萄球菌是主要污染菌.     

A Simple, Inexpensive Metabolism Cage for Small Mammals

By utilizing ordinary laboratory equipment and a spherical feces-urine separator, a simple, inexpensive metabolism cage for small mammals can be constructed. A hardware cloth animal cage over a cylindrical battery jar containing a spherical feces-urine separator affords the following advantages not commonly found in commercial metabolism cages: 1) complete separation of feces and urine through minimal contact, 2) minimal evaporation of urine due to proximity of storage vessel and lack of exogenous air currents, and 3) extremely low cost of less than five dollars. The metabolism cage is designed to allow measurement of fluid intake, and to separate and collect feces and urine for numerous qualitative and quantitative determinations. In addition, the metabolism cage permits observation of the animal, feces, and urine at all times, is readily cleaned or sterilized, and is easily fashioned from common laboratory equipment.     

犬轮状病毒单克隆抗体制备及胶体金试纸条检测方法的建立

【目的】研发犬轮状病毒(Canine ratavirus, CRV)免疫学诊断试剂,制备并鉴定CRV特异性单克隆抗体,建立可用于检测CRV的胶体金免疫层析法。【方法】以CRV临床分离株SL006株为免疫原免疫BALB/c雌性小鼠,用杂交瘤细胞法进行细胞融合,通过间接免疫荧光法(IFA)筛选可稳定分泌CRV单克隆抗体的杂交瘤细胞,并制备腹水,亲和层析法进行纯化获得单克隆抗体。对获得的单克隆抗体进行鉴定,经条件优化建立胶体金试纸条检测方法,对试纸条的灵敏度、特异性、重复性和应用情况进行评价。【结果】获得了4株杂交瘤细胞,分别命名为2C12、4A5、1H3、5F3,IFA效价分别为1∶6 400、1∶12 800、1∶1 600和1∶3 200;重链亚类分别为IgG2b、IgG2a、IgG1和IgG2a,轻链亚类均为kappa;制备的胶体金试纸条检测线包被单克隆抗体4A5,对照线包被羊抗鼠IgG,金标垫包被胶体金标记的单克隆抗体2C12,对CRV病毒液的检测灵敏度为10^4.2 TCID₅₀/mL,检测犬源其他病毒犬细小病毒(Canine parvo...     

猪轮状病毒VP4基因重组腺病毒的构建及抗体水平评价

【目的】构建猪轮状病毒（Porcine rotavirus,PoRV）流行株PoRV G9P[23]型的VP4基因重组腺病毒,为开发PoRV候选基因工程疫苗奠定基础。【方法】参考GenBank中流行株PoRV G9P[23]型的VP4基因序列（登录号：MH898990.1）合成PoRV VP4基因,将得到的目的基因与腺病毒穿梭载体pAdTrack-CMV进行重组,转化大肠杆菌Top10感受态细胞,构建腺病毒穿梭载体pAdTrack-CMV-VP4;腺病毒穿梭载体经过Pme Ⅰ内切酶线性化处理后与含有腺病毒骨架pAdEasy-1的大肠杆菌BJ5183感受态细胞进行同源重组获得重组质粒pAd-VP4,对重组质粒进行Pac Ⅰ酶切鉴定,并转化大肠杆菌DH5α感受态细胞。将重组质粒转染HEK293A细胞获得重组腺病毒rAd-VP4,对该重组腺病毒进行扩大培养并测定重组腺病毒的半数组织培养感染剂量（TCID₅₀）;通过RT-PCR检测其体外表达情况,Western blotting检测其反应原性;将制备的重组腺病毒用不同病毒滴度和不同免疫次数对小鼠进行腹腔免疫,收集血清通过ELISA法测定IgG抗体水平。【结果】RT-PCR扩增出1条大小为2 343 bp的rAd-VP4重组腺病毒条带,测序结果正确,表明重组腺病毒rAd-VP4构建成功,测得rAd-VP4病毒滴度为10^6.5 TCID₅₀,Western blotting结果表明,重组腺病毒rAd-VP4在蛋白水平上得到了正确表达,蛋白的分子质量约为87 ku。小鼠IgG抗体检测结果表明,在用10^6.5 TCID₅₀ rAd-VP4免疫后的第35和42天,小鼠血清中的IgG抗体水平显著高于10^5.3 TCID₅₀ TGE-PED-PRV三联活疫苗IgG抗体水平（P<0.05）;10^6.5 TCID₅₀ rAd-VP4在免疫后第35和42天产生的抗体水平显著高于10^5.5和10^4.5 TCID₅₀ rAd-VP4（P<0.05）,而10^5.5 TCID₅₀ rAd-VP4在免疫后第28天产生的抗体水平显著高于10^6.5和10^4.5 TCID₅₀ rAd-VP4（P<0.05）。10^6.5 TCID₅₀ rAd-VP4的2次免疫和3次免疫产生的IgG抗体在不同免疫时间均差异不显著（P>0.05）。【结论】本研究成功构建了重组腺病毒rAd-VP4,其病毒滴度为10^6.5 TCID₅₀。10^6.5和10^5.5 TCID₅₀ rAd-VP4分别在第42和28天产生较高的IgG抗体水平,2次免疫和3次免疫对产生IgG抗体水平均无显著影响。试验结果可为开发PoRV重组腺病毒候选疫苗提供参考。     

动物园的科学发展观与市场化选择

本文从动物园的发展实践中，提出动物园的科学发展观；从发展机遇、体制创新、市场定位、功能发挥、文化建园、开拓市场、科学管理和依法经营8个方面论述了市场经济对我国动物园提出的要求。     

Intraperitoneal Insemination in Mammals: A Review

This review focuses on factors associated with the development of intraperitoneal insemination in mammals. Findings to date indicate that fertility improves as the sperm cell concentration rises, but that the optimal sperm number differs in each species. Sperm washing before intraperitoneal insemination favours fertility. Peritoneal fluid shows a variable effect on spermatozoa, depending on the hormonal status of the female. The optimal time for insemination appears to be just prior to ovulation. The technique may be performed either through the abdominal or the vaginal wall. Verification of sperm deposition in the proximity of the ovaries improves fertility rates. Although associated with some risk of infection and an immune reaction against spermatozoa, the intraperitoneal technique rarely gives rise to severe anaphylactic shock, peritonitis, adhesion formation and the production of anti‐sperm antibodies and these complications may be prevented by adequate sperm pretreatment and antibiotic therapy. The success of intraperitoneal insemination in humans, with results comparable with those of intrauterine insemination in the treatment of infertility, suggest the potential use of this technique in domestic mammals, especially in those in which intrauterine insemination poses practical difficulties. Some of the methods applied in human intraperitoneal insemination, such as confirming the position of the needle in the peritoneal cavity, and sperm pre‐treatments might also improve results in domestic species. Conversely, the use of the animal model should help to develop some aspects of this technique in humans.     

Survey of Zoonotic Dermatoses in Client‐Owned Exotic Pet Mammals in Southern Italy

Several ‘exotic’ mammalian species (e.g. rabbits, rodents, ferrets and hedgehogs) live in close proximity to humans as companion pets. Skin diseases (SD) are frequent causes of morbidity in exotic pet mammals, and most of those SDs have a zoonotic potential. The purpose of this study was to determine the frequencies and types of zoonotic dermatosis (ZD) in client‐owned, exotic pet mammals in Southern Italy. Six‐hundred and fifty‐five medical records of exotic pet mammals examined between 2011 and 2012, across twenty private practice veterinary clinics around the Naples area (Italy), were retrospectively evaluated and screened for animals diagnosed with SDs (rabbits n = 455, guinea pigs n = 93, ferrets n = 64, hedgehogs n = 19, chinchillas n = 13 and rats n = 11). The records of animals diagnosed with SD, whose causative agents had a zoonotic potential, were selected for analysis. The Mann–Whitney independent test was used for statistical analysis. A P value ≤0.05 was considered statistically significant. Eighty‐two records (12.5%) of animals with ZD were identified. Of those, 56.1% (46/82) were affected by fungal infections and 42.7% (35/82) by parasitic infections. No zoonotic bacterial or viral infections were diagnosed. Dermatophytosis was significantly diagnosed more frequently in younger animals. The results of this survey indicate that exotic pet mammals may serve as active carriers for many highly contagious pathogens with zoonotic potential. Awareness and vigilance by the veterinary practitioner is crucial in the prevention of occurrences of ZDs. Children frequently come in close contact with exotic pets. To prevent the unplanned transmission of pathogen from pet to human, an active routine screening examination and preventative treatments are strongly recommended for every newly purchased pet mammal.     

应用单克隆抗体酶联免疫斑点试验检测抗传染性喉气管炎抗体

本文提出以硝酸纤维膜作为固相载体，辣根过氧化物酶标记的抗传染性喉气管炎病毒（ＩＬＴＶ）的单克隆抗体（ＭｃＡｂ），邻苯二胺为底物，检测ＩＬＴＶ抗体的免疫斑点试验方法（Ｄｏｔ－ＥＬＩＳＡ）。该方法较常规方法敏感，特异、省时。该方法的建立为鸡群疫病监测，免疫鸡群抗体水平的检测提供了新的可靠方法。     

猪A群轮状病毒的分离与鉴定

目前国内对猪轮状病毒的研究较少,作者旨在对猪A群轮状病毒进行分离与鉴定,为后续猪轮状病毒致病机理和分子生物学特性研究奠定基础.用胰蛋白酶处理RT-PCR检测猪A群轮状病毒病原阳性的临床腹泻粪样,然后接种长成单层的MA-104细胞,进行病毒分离传代.再对分离毒株进行常规RT-PCR鉴定和电镜观察,并对分离毒株的VP6、VP7和VP8基因进行测序及序列分析.结果成功分离猪A群轮状病毒TM-a株,经序列同源性分析发现,与TM-a株VP6、VP7和VP8基因同源性最高的毒株分别为中国北京人源LL3354株、印度人源RMC321株和日本野猪源GUB-71株,其相似性为96.0％、95.1％和97.0％.该TM-a株轮状病毒不同基因表现出与人源轮状病毒和猪源轮状病毒的高度同源性,推测猪A群轮状病毒可能在人畜间传播,发生基因重组现象.     

A Quantitative Flurometric Assay for the Measurement of Antibody to Pasteurella haemolytica in Cattle

A rapid, simple fluorometric method is described for measuring antibody to Pasteurella haemolytica in sera of cattle. Various antigen preparations were compared for the test including live, formalin-killed and phenol-killed P. haemolytica. A preparation composed of formalin-killed organisms from a 22 hour culture gave consistent results and was used in the studies. The test was reproduciable with percent coefficients of variation for fluorescent signal unit values on ten or more replicate samples ranging from 5.7 to 28.0. Sera from calves vaccinated by aerosol exposure to live P. haemolytica had up to a five-fold increase in antibody titer as measured by the flurometric method test during a 21 day period. Fluorometric method titers were comparable to those obtained by the indirect bacterial agglutination test. There was no seroconversion to P. haemolytica in calves vaccinated by aerosol exposure of P. multocida. The major advantages of the fluorometric method test over conventional methods are that the assay does not require serial dilutions of serum samples and thus limits time and effort to determine antibody titers.     

动物园动物的应激行为与动物福利管理

动物园动物是人工驯养的动物,动物的应激行为表现是动物与饲养管理人员、游人之间和与笼舍环境的应激,是人为造成的。分析广州动物园动物应激行为表现,与广州动物园动物的福利水平,提出动物园动物福利管理思路和方法。减少动物园动物的应激行为,在饲养管理中定人、定点、定时、定量。在动物繁殖期分笼,设立专门的产房和育幼室。应用动物行为丰容技术和动物园笼舍建设生态化。让动物康乐生活,防疫好,疾病少,寿命长,合理繁殖。     

广州动物园草食动物饲用木本植物的筛选

象草是广州动物园草食动物的主要青饲料,但在冬季和早春季节象草供应量不足,且营养价值明显下降。洋蹄甲、山指甲和小叶榕是广州常见的3种常绿木本植物,但没有作为饲料的报道。我们通过动物的喜食程度比较,饲养效果观察,植物冬季营养成分分析以及抗营养因子胰蛋白酶抑制剂活性的测定,发现这3种植物适宜广州动物园草食类动物的食用,特别是在冬季和早春可以用来替代部分象草,以弥补象草供应的不足。