从云豹分离的一株猫泛白细胞减少症病毒的特性

从病死的云豹体内分离到1株猫泛白细胞减少症病毒,进行了鉴定,研究了它的血凝特性和对猫与水貂的致病力,分析了它与猫泛白细胞减少症病毒参考毒株(FNF-8)抗原的相关性。 用中国兽药监察所提供的猫肾传代细胞(FK)进行病毒的分离鉴定。同步接种和异步接种测得病毒的TCID50均为4.8/ml。病毒经乙醚、酸(pH3.0)、热(56℃1小时)     

虎源猫泛白细胞减少症病毒的分离鉴定

利用细胞培养方法从中国某虎园送检的腹泻虎肠内容物中分离出1 株细小病毒(TPV/ HT - 69),经系统的形态学、理化学、血清学试验、人工感染试验、PCR 扩增和VP2 基因序列分析,符合猫泛白细胞减少症病毒特征,证明该毒株为猫泛白细胞减少症病毒强毒。     

犬、猫、貂细小病毒通用快速诊断盒的研制

本文报道,由犬细小病毒灭活抗原、特异抗体、戊二醛醛化猪红细胞及微量血凝板、稀释棒等组成的犬、猫、貂细小病毒通用快诊盒,可用于犬细小病毒性肠炎、猫泛白细胞减少症和水貂肠炎的特异诊断,适合于基层和野外应用,并可在接到病料后4小时内报告结果,有关试剂的有效期在一年以上。     

四种动物病毒的细胞培养及血凝检测的比较研究

四种动物病毒的细胞培养及血凝检测的比较研究李天宪,赵林,罗怡珊,冯锋（中国科学院武汉病毒研究所,武汉４３００７１）关键词细小病毒,细胞培养,细胞病变,血凝试验云豹肠炎病毒（ＬＰＶ）、水貂肠炎病毒（ＭＥＶ）、犬肠炎病毒（ＣＰＶ）和猫泛白细胞减少症病毒（...     

虎源猫泛白细胞减少症病毒灭活疫苗的制备及初步应用

摘要：【目的】研制出虎源猫泛白细胞减少症病毒灭活疫苗并对其应用效果进行评价。【方法】以FPV-HLJ为种毒,按1×10-2接毒量,采用同步接毒的方法接于猫肾传代细胞系F81株,于37 ℃静置培养,待细胞病变（CPE）达到75 %以上时,进行收毒。病毒悬液经甲醛灭活24 h,加入佐剂氢氧化铝胶,制备成FPV-HLJ细胞培养灭活疫苗。【结果】皮下接种2月龄非免疫家猫,结果显示,实验组免疫猫FPV HI抗体水平随免疫次数的增加而呈上升趋势,三免后FPV HI抗体效价为1:1024～1：2048,且免疫猫能抵抗FPV强毒攻击,具有100 %的存活率。随后将该苗接种2月龄左右幼虎,三免后抗体效价多数能达到1：1024。【结论】表明此灭活苗可产生较为理想的免疫效果。     

犬细小病毒单克隆抗体的制备及其与免疫血清在血凝抑制中的比较

将用犬细小病毒免疫的ＢＡＬＢ／ｃ小鼠的脾细胞与Ｓｐ２／０骨髓瘤细胞在聚乙二醇作用下融合,经筛选、克隆,得到６株稳定分泌犬细小病毒单克隆抗体的杂交瘤。用杂交瘤腹水作血凝抑制试验,检测分别含有犬细小病毒、猫泛白细胞减少症病毒和水貂肠炎病毒的标本,及ＣＰＶ感染犬粪便标本,并与免疫血清作对比。结果表明,用单克隆抗体作血凝抑制试验,比用免疫血清作具有特异性高、操作简便省时等优点,而二者敏感性一致。     

犬细小病毒:从起源到进化

犬细小病毒(Canine parvovirus,CPV-2)首次分离于1978年,被认为是由遗传关系相近的猫泛白细胞减少症病毒(FPLV)或其他肉食兽细小病毒(FPLV-like virus)跨宿主感染犬产生的新病原.其感染能引起新生犬急性心肌炎或幼犬出血性肠炎,造成较高的发病率和死亡率.该病原爆发后短短一年时间内即广泛流行到世界各地.随着CPV-2对宿主的适应和变异,新抗原变异型(CPV-2a、CPV-2b和CPV-2e)不断产生并在世界各地逐步替代了CPV-2的流行.伴随CPV-2抗原变异的同时,其对宿主(犬、猫)嗜性、毒力等生物学特性也随之改变.本文综述了CPV-2过去30多年在世界流行和变异情况,并探讨了基因变异在病毒跨宿主传播中的意义.     

敏感性不同细胞系交替选育获得超高增殖率细小病毒株

迄今为止未见国际上有组织培养水貂肠炎病毒(MEV)、猫泛白细胞减少症病毒(FPLV)、犬细小病毒(CPV)的TCID_(50)高于log_(10)7.0或MEV的HA高于32~或CPV的HA高于4096~的报道。又据Goto(1986)报道,TCID_(50)为log10~(4-6)的CPV比MEV。FPLV的HA高20多倍,故HA为4096~的CPV液比HA为32~的MEV液中病毒含量还低。由此可见,PV培养效价低是目前国际上急待解决的重大问题。     

猫肾F81传代细胞中犬细小病毒原位PCR检测方法

目的建立在猫肾F81细胞中犬细小病毒原位PCR的检测方法.方法在猫肾F81细胞上感染犬细小病毒,设计特异性引物,用直接原位PCR法在染毒12h,24h,48h细胞片上检测出犬细小病毒,并与常规免疫组化的方法进行了比较.结果在染毒48h的细胞片上,用阳性记分法将两种检测方法得到的阳性细胞进行统计比较,差异极显著(P＜0.001),用原位PCR法所得出的阳性率高.结论原位PCR法检测犬细小病毒具有敏感性高和组织定位的优点.     

北京地区猫疱疹病毒Ⅰ型的分离及鉴定

了解北京地区猫疱疹病毒Ⅰ型(FHV-1)毒株特点,为猫疱疹病毒Ⅰ型的进一步研究及对猫的感染预防控制奠定基础。经特异性PCR检测为FHV-1阳性临床猫眼鼻拭子,经过处理,接种CRFK细胞,通过细胞培养分离病毒。通过间接免疫荧光试验、电镜形态观察、病毒毒力测定及基因序列分析等对分离株进行鉴定。PCR检测FHV-1阳性的14只猫眼鼻拭子接种CRFK细胞,有6只猫样本能使CRFK细胞发生圆缩、拉网集聚等病变;6个样本接种的CRFK病变细胞滴片进行免疫荧光试验,均产生绿色荧光反应;经传代培养,有5株分离株能够稳定传代,其中有2株弱毒,3株强毒;电镜下5株分离株均可见直径100~160nm的球型病毒粒子;5株分离株与FHV-1标准株C-27及2个不同公司来源疫苗株gB、gD基因核苷酸比较同源性均为99%以上。实验室成功分离保存5株能稳定传代的FHV-1毒株,5株分离株与FHV-1标准株及2个疫苗株亲缘关系很近。     

老虎感染猫传染性鼻—结膜炎病毒的研究

用猫肾传代细胞从发病虎的病料中分到了一株杯状病毒粒子样病毒。该病毒大小为３５～３９ｎｍ、无囊膜、在胞浆中病毒前体呈晶格状排列,抗乙醚、不能被５－ＩＵＤＲ所抑制,能被来源于标准疫苗的猫传染性鼻－结膜炎病毒（或杯状病毒Ｆｅｌｉｎｅ ｃａｌｉｃｉｖｉｒｕｓ,ＦＣＶ）抗血清所中和,人工感染猫出现典型的ＦＣＶ感染症状。ＲＴ－ＰＣＲ能扩增出与设计值相符的电泳带,ＰＣＲ产物直接测序结果与Ｇｅｎｅｂａｎｋ发表的Ｆ     

Cytotoxic effects induced by interferon-ω gene lipofection through ROS generation and mitochondrial membrane potential disruption in feline mammary carcinoma cells

Progress in comparative oncology promises advances in clinical cancer treatments for both companion animals and humans. In this context, feline mammary carcinoma (FMC) cells have been proposed as a suitable model to study human breast cancer. Based on our previous data about the advantages of using type I interferon gene therapy over the respective recombinant DNA derived protein, the present work explored the effects of feline interferon-ω gene (fIFNω) transfer on FMC cells. Three different cell variants derived from a single spontaneous highly aggressive FMC tumor were successfully established and characterized. Lipofection of the fIFNω gene displayed a significant cytotoxic effect on the three cell variants. The extent of the response was proportional to ROS generation, mitochondrial membrane potential disruption and calcium uptake. Moreover, a lower sensitivity to the treatment correlated with a higher malignant phenotype. Our results suggest that fIFNω lipofection could offer an alternative approach in veterinary oncology with equal or superior outcome and with less adverse effects than recombinant fIFNω therapy.     

A serological survey of common feline pathogens in free-living European wildcats (<Emphasis Type="Italic">Felis silvestris</Emphasis>) in central Spain

Twenty-five serum samples of 22 free-living European wildcats (Felis silvestris) captured from 1991 to 1993 in central Spain were tested for evidence of exposure to seven feline pathogens. All the wildcats but one (95.4%) presented evidence of contact with at least one of the agents (mean = 2.2). Contact with feline leukemia virus (FeLV) was detected in 81% of the wildcats (antibodies, 77%; antigen p27, 15%). Antibodies to feline calicivirus (FCV, 80%), feline herpesvirus (FHV, 20%), feline parvovirus (FPV, 18%), and Chlamydophila sp. (27%) were also detected. Analyses were negative for feline immunodeficiency virus and feline coronavirus. The probability of having antibodies to FPV was inversely related with the concentration of serum cholesterol and with a morphometric index of body condition. Similarity in the composition of antibodies against disease agents (number and identity of detected and undetected antibodies) was significantly higher in pairs of female wildcats than in pairs of males or heterosexual pairs, suggesting that females had a more homogeneous exposure to pathogens. Seroprevalence for FHV was higher in males than in females. Antibodies to FHV and Chlamydophila sp. were more frequent in winter than in other seasons. In addition, the mean similarity of the pathogen community between pairs of serum samples was higher if both wildcats were caught during the same season than if they were not. Mean similarity was lowest when serum samples obtained in winter were compared with those from spring or summer. The results suggest that some agents probably had a reservoir in domestic cats and may cause some undetected morbidity/mortality in the studied wildcat population, whereas others, such as FeLV and FCV, may be enzootic.     

The Relationship Between Coat Color and Aggressive Behaviors in the Domestic Cat

The authors explored a possible relationship between coat color and aggressive behaviors in the domestic cat. This study used an Internet-based survey to collect information on coat color, affiliative behaviors toward cats/humans, agonistic behaviors toward cats/humans, other “problem” behaviors, and cat and guardian demographic data. A total of 1,432 cat guardians completed the online survey; after exclusions based on study protocol, data analysis included 1,274 completed surveys. Guardians reported sex-linked orange female (tortoiseshells, calicos, and “torbies”), black-and-white, and gray-and-white cats to be more frequently aggressive toward humans in 3 settings: during everyday interactions, during handling, and during veterinary visits. Kruskal-Wallis 1-way analysis of variance was used to compare possible differences between the 2 sexes and among different coat colors. Analyses of aggression due to handling, as well as aggression displayed during veterinarian visits, showed little difference among coat colors in these settings.     

表达猫瘟热病毒VP2蛋白重组腺病毒的构建及其免疫原性研究

为构建能表达FPV VP2蛋白的重组犬2型腺病毒(CAV-2)载体.首先用PCR方法从FPV GT-2株细胞培养物中扩增出了VP2蛋白基因,将其克隆到真核表达质粒pVAX1中构建了含有FPV vp2基因的表达盒(CMV-VP2-PolyA),将该表达盒酶切后定向克隆到含有CAV-2 E3区的穿梭质粒pVAX△E3中,构建出pVAX△E3VP2.用Sal I+Nru I双酶切pVAX△E3VP2,回收含有目的基因表达盒部分,将其定向克隆入含有CAV-2全基因组的骨架质粒pPoly2-CAV-2中,构建了重组质粒pCAV-2-FPV-VP2.Cla I+Asc I酶切pCAV-2-FPV-VP2释放出重组基因组,以此转染MDCK细胞,获得了重组病毒CAV-2-VP2.该重组病毒能使MDCK细胞产生腺病毒样细胞病变.Western blot检测证实,该重组病毒能表达具有免疫学活性的VP2蛋白.该重组病毒可以有效地诱导免疫猫产生抗FPV和CAV-2抗体.本实验表明该重组病毒有可能成为一个FPV的疫苗株.     

表达猫瘟热病毒VP2蛋白重组腺病毒的构建及其免疫原性研究

为构建能表达FPV VP2蛋白的重组犬2型腺病毒(CAV-2)载体。首先用PCR方法从FPV GT-2株细胞培 养物中扩增出了VP2蛋白基因,将其克隆到真核表达质粒pVAX1中构建了含有FPV vp2基因的表达盒(CMV- VP2-PolyA),将该表达盒酶切后定向克隆到含有CAV-2 E3区的穿梭质粒pVAX△E3中,构建出pVAX △E3VP2。用Sal I Nru I双酶切pVAX△E3VP2,回收含有目的基因表达盒部分,将其定向克隆入含有CAV-2 全基因组的骨架质粒pPoly2-CAV-2中,构建了重组质粒pCAV-2-FPV-VP2。Cla I Asc I酶切pCAV-2-FPV- VP2释放出重组基因组,以此转染MDCK细胞,获得了重组病毒CAV-2-VP2。该重组病毒能使MDCK细胞产生 腺病毒样细胞病变。Western blot检测证实,该重组病毒能表达具有免疫学活性的VP2蛋白。该重组病毒可以有 效地诱导免疫猫产生抗FPV和CAV-2抗体。本实验表明该重组病毒有可能成为一个FPV的疫苗株。     

Low temperature compartment formation in feline immunodeficiency virus-infected and uninfected feline kidney cells

Summary This study was to determine if feline immunodeficiency virus (FIV)-infected and uninfected Crandall feline kidney (CRFK) cells exhibited a low temperature (16°C) block in membrane trafficking between transitional endoplasmic reticulum and Golgi apparatus represented by intermediate compartment formation. Cells were cultured at different temperatures and membrane changes involving the Golgi apparatus and Golgi apparatus-associated membrane structures were monitored by electron microscopy and quantitated. With 30 min of incubation, membranes of the Golgi apparatus stack increased in amount at temperatures of 16°C and below compared to temperatures above 18°C. The increase was greatest along the major polarity axis as evidenced by an increased stack height. Neither the number of cisternae per stack nor the average stack diameter (width) was affected by temperature. The response was maximal between 15 and 30 min of low temperature treatment of the cells. Results with cells infected and uninfected with feline immunodeficiency virus were similar. The increase in stack height was due primarily to an increase of membranes at the cis face (cis Golgi apparatus network). At 18°C, membranes of the trans Golgi apparatus network accumulated suggesting that import from the cis Golgi network could proceed at this temperature, whereas exit from the trans Golgi network was still at least partially blocked. Also increased at 16°C and below were numbers of transition vesicles in the space between the Golgi apparatus and the transitional endoplasmic reticulum associated with the cis Golgi apparatus face. The results suggested interruption of the orderly flux of membranes into the Golgi apparatus at 16°C and below. Moreover, the block appeared to be reversible. Upon transfer from 16°C to 37°C, there was a time-dependent decrease in the accumulations of cis compartment membrane accompanied by a corresponding equivalent increase in the membranes of the trans Golgi apparatus compartment.     

Long-term melatonin treatment prolongs interestrus, but does not delay puberty, in domestic cats

The objective was to assess the efficacy and safety of long-term administration of melatonin (either as an implant or given orally) on interestrus intervals in domestic cats. Additionally, the effect of melatonin implants on puberty postponement was studied. For these purposes, two randomized controlled trials were conducted. In the first, 68 interestrus intervals (in 28 postpubertal queens) were studied, and in the second, 32 prepubertal female cats were used. During anovulatory interestrus intervals (27 ovulatory interestrus intervals were excluded), postpubertal cats were assigned to the following three treatments: melatonin implant 18 mg/cat SC (n = 17; MEI); melatonin tablets, 4 mg/cat/d orally until the onset of estrus (n = 12; MEO); or control (n = 12; CTL). Prepubertal females were randomly assigned to the following three treatments: melatonin 18 mg/cat sc implants at either 1.9 ± 0.3 kg (MEI-A; n = 12) or 1.5 ± 0.3 kg (MEI-B; n = 10) body weight; or control (CTL; n = 10). Interestrus intervals in postpubertal MEI, MEO, and CTL groups were 63.8 ± 5.4, 63.0 ± 5.3 and 19.2 ± 1.4 d (P < 0.05), respectively. In these groups, intervals between onset of treatment and the first estrus cycle were 51.0 ± 4.7, 50.0 ± 6.1, and 12.6 ± 1.1 d (P < 0.05). In the second experiment, neither age (MEI-A: 232.4 ± 10.5, MEI-B: 208.6 ± 13.0 and CTL: 192.4 ± 20.1 d; P > 0.1) nor body weight (P > 0.1) at puberty differed among groups. None of the cats in either study had clinically apparent side effects. We concluded that long-term melatonin treatment of domestic cats slightly prolonged interestrus intervals, but did not postpone puberty.