大熊猫犬冠状病毒的分离与鉴定

以犬肾传代细胞(MDCK)从1只病死大熊猫肝脏中分离到1株病毒,命名为DXMV。电镜观察磷钨酸负染的病毒细胞培养物，发现冠状病毒样粒子，超薄切片上可见细胞浆内病毒包涵体。理化学和生物学鉴定结果表明，DXMV核酸类型为RNA，对氯仿、乙醚敏感，可耐受1％胰蛋白酶的作用，具有一定的耐酸性和耐热性，可在MDCK、FCWF、F81细胞中增殖，无血凝性和血吸附特性。血清中和试验结果表明，DXMV可被犬冠状病毒阳性血清中和。采用犬冠状病毒间接荧光抗体法染色DXMV细胞飞片，可在细胞浆内见到特异性荧光。以冠状病毒通用引物和犬冠状病毒特异性引物，可从大熊猫肝脏和不同代次的病毒细胞培养物中扩增出与预期值251bp和515bp大小相同的核苷酸片段。由此说明，该病毒为犬冠状病毒，大熊猫可被犬冠状病毒感染。     

简述犬的冠状病毒病

当前出现的人类新型冠状病毒(COVID-19)病给整个社会生活和人民健康带来了巨大的影响。犬也会患冠状病毒病,犬冠状病毒病会不会传染给人呢?于此,就犬冠状病毒病做一个简要的论述。一、冠状病毒病的前世今生冠状病毒按病毒学分类属于尼多病毒目、冠状病毒科、冠状病毒亚科。之所以称为冠状病毒是因为其电镜外形类似皇冠样。     

从猝死症犬心肌分离出冠状病毒

应用电镜观察，在猝死犬的心肌中发现典型的冠状病毒粒子。用牛肾传代细胞（ＭＤＢＫ）分离培养该病毒获得成功，电镜负染色法跟踪观察，每一代细胞培养液中均观察到典型的冠状病毒粒子。其病毒形态呈圆形和椭圆形为主的多形性，直径７０～１６０ｎｍ，有囊膜，囊膜表面有间距较宽（长约２０ｎｍ）的突起，突起顶端膨大呈鼓槌样，负染色法观察时呈现花冠样结构。用第５代接毒培养的细胞作电镜超薄切片观察，在细胞质的空泡中发现有数量不等的典型冠状病毒粒子，即病毒包涵体。经鉴定为ＲＮＡ病毒，对乙醚敏感，３７℃７２ｈ可失去活性。在ｐＨ３的条件下，３７℃５ｈ仍保持一定活力     

犬冠状病毒研究进展

2020年2月,我国香港报道了新冠肺炎（COVID-19）患者家养的宠物犬感染了新型冠状病毒（SARS-CoV-2）,这引起了人们对犬冠状病毒的关注。为有助于人们全面了解犬冠状病毒,做好犬冠状病毒病防治,本文从病原学与流行病学、临床症状以及诊断和防治措施等方面进行了综述。犬冠状病毒在犬群中广泛存在,主要包括α冠状病毒属的犬肠炎冠状病毒和β冠状病毒属的犬呼吸道冠状病毒。犬冠状病毒致病性不强,引起的临床症状通常较轻,但对幼犬或与犬细小病毒、犬瘟热病毒等发生混合感染时,会引起较严重症状,且易发生突变,产生新的变异毒株,具有致病性增强的风险。目前,犬冠状病毒检测方法主要包括病原学检测、血清学检测以及RT-PCR、qRT-PCR、纳米PCR等分子生物学检测。犬冠状病毒感染的防治主要通过疫苗免疫以及科学饲养管理、卫生消毒和对症治疗等综合措施。虽然犬可感染SARS-CoV-2,但尚不清楚感染犬是否可将病毒再传染给其他动物或人类。因此,应加强犬冠状病毒的病原生态学研究和监测预警。     

犬冠状病毒YS1株人工感染犬试验

使用从腹泻犬体内分离获得的犬冠状病毒ＹＳ１第５代ＣＲＦＫ细胞培养物,以不同剂量人工感染５０只２￣３月龄健康幼犬 ,１４ｄ内计有１６只犬发病,其中１４只耐过,２只死亡,发病率为３２％,死亡率为１２．５％,患犬粪便经电镜检查和ＲＴ－ＰＣＲ检测,前者见有ＣＣＶ样病毒,后者扩增出ＣＣＶ特异核酸片段。４８只试验犬血清对该病毒的ＳＮ抗体效价随接毒量的不同分别平均为１：３１０、１：２４５、１：１８５、１：１１５     

犬冠状病毒病的诊断与控制

犬冠状病毒病是由犬冠状病毒引起的一种犬的急性胃肠道传染病。它以传播迅速、致死率高,发病后呈不同程度的胃肠道炎症为特征。1病原犬冠状病毒(CCV)是冠状病毒科冠状病毒属中的一个新成员。电镜下病毒呈圆形(直径80～100nm),或椭圆形(长180～200nm,宽75～80nm),有囊膜,表面有长约20nm的纤突,纤突末端呈球状,含单股RNA。CCV对热、紫外线、福尔马林、石炭酸、乙醚、氯仿及去氧胆酸盐、来苏尔、0.1%过氧乙酸、1%克辽林等敏感。病毒能在许多犬的原代细胞或传代细胞上生长。病毒主要存在于感染犬的肠内容物中和肠上皮内,在肠系膜淋巴结及其…     

1例犬冠状病毒病并发胰腺炎的诊治

1只1岁比熊犬进食后多次呕吐,排便先硬后稀,精神沉郁,通过临床检查、试剂盒检测及血常规检查进行诊断,初步确诊为犬冠状病毒病,对症治疗3 d,呕吐症状仍存在,遂进行血常规检查、生化检查及cPL检测,确诊该犬还患有胰腺炎,并及时调整治疗方案。结果表明:患犬为犬冠状病毒病并发胰腺炎,根据该犬的实际情况采取对症治疗和对因治疗的措施,患犬预后良好。     

犬猫的冠状病毒病与“非典”无关

目前,提起“非典型性肺炎”令人紧张异常。由于报道说“人体中以前没有发现过冠状病毒”、“冠状病毒可能来源于动物”等说法,使养动物的人们感到困惑,已有许多犬、猫的主人打电话到中国农业大学动物医院,询问动物的冠状病毒事宜,在此,作为临床兽医工作者,我认为有必要将犬、猫的冠状病毒病与这次人的“非典”作个对比,并将犬、猫的冠状病毒病加以说明。 犬的冠状病毒病不是什么新病。至今为止,世界各国的犬冠状病毒仅感染犬,从来没有发生过感染犬的主人或者兽医的情况,这是不容置疑的事实。 犬的冠状病毒感染犬后引起犬的胃肠炎,特征包…     

一例犬冠状病毒病的病例报告

犬冠状病毒病是由犬冠状病毒引起的一种临床上以呕吐、腹泻、脱水及易复发为特性的高度接触性传染病。该病对幼犬危害严重,如诊断和治疗不及时,容易导致犬只死亡。就一例犬冠状病毒病的临床诊断及治疗方案进行总结,为提高该病的诊断及治疗水平提供参考。     

犬冠状病毒感染

犬冠状病毒感染是由犬冠状病毒引起犬的一种以急性胃肠炎为主要症状的高度接触性传染病 ,对养犬业危害严重。本文对近年来该病的病原、流行病学、诊断和防治等方面的国内外研究进展进行了综述。     

Comparison of the prevalence of enteric viruses in healthy dogs and those with acute haemorrhagic diarrhoea by electron microscopy

Objectives: To evaluate prevalence of enteric viruses in healthy dogs and to compare it with prevalences in dogs with acute haemorrhagic diarrhoea. Methods: Faecal samples were collected from 200 healthy dogs and examined by electron microscopy for presence of viral particles. Data were compared with viral prevalences that had been determined retrospectively by electron microscopy for 936 dogs with acute haemorrhagic diarrhoea. Results: There were significantly more negative faecal samples among the healthy dogs (82.0 per cent) compared with 55.8 per cent in dogs with acute haemorrhagic diarrhoea (P<0.001). With a prevalence of 17.5 per cent, significantly more healthy dogs were shedding coronavirus compared with 11.6 per cent in dogs with acute haemorrhagic diarrhoea (P=0.034). Parvovirus was only detected in one healthy dog (0.5 per cent), thus with a prevalence that was significantly lower than 16.0 per cent detected in the dogs with acute haemorrhagic diarrhoea (P<0.001). Paramyxovirus was not found in any of the healthy dogs but was found in 9.3 per cent of dogs with acute haemorrhagic diarrhoea (P<0.001). Clinical Significance: Results suggest that shedding of parvovirus and paramyxovirus is strongly associated with acute haemorrhagic diarrhoea. However, coronavirus seems to be even more prevalent among healthy dogs, raising the need for further studies to investigate the strain-associated pathogenicity of this virus.     

Electron microscopic detection rate of enteral viruses in diarrhea of dogs, cats, calves, swine and foals in the year 1988--electron microscopic study results

During 1988 fecal and gut samples of 641 dogs, 198 cats, 576 calves, 108 piglets and 64 foals with diarrhoea were investigated for virus infections by electron microscopy. In samples of dogs and cats parvovirus was detected at a proportion of 21.9% and 16.7%, respectively; rotavirus alone or together with coronavirus was found only in 0.3-1.5% of the specimens. In samples of calves rotavirus, as well as coronavirus dominated with a detection rate amounting to 17.4% and 26.6% respectively (including 4.5% of mixed infections); parvovirus was present in a ratio of 0.5%. Specimens of piglets mainly contained coronavirus (25.0%), and in lower percentages rotavirus (2.8%), rota- and coronaviruses (0.9%) and parvovirus (0.9%). In feces of foals rotavirus was detected in 6.3% and particles resembling picornavirus in 4.7% of cases. Not identifiable virus particles resembling corona-or picornaviruses were rarely found (between 0.6-2.5) also in specimens of the other animal species.     

Viruses and virus-like particles in the faeces of dogs with and without diarrhoea

Negative staining electron microscopy was used to identify viruses in 157 normal and 29 diarrhoeal faecal samples collected from 156 dogs admitted to an animal shelter during an 8 month period (March to October) in 1982. Seven distinct viral types were detected: 21-26 nm parvovirus-like particles, 28-31 nm astrovirus-like particles, a previously undescribed 34-35 nm "round" virus particle, coronavirus, coronavirus-like particles ( CVLP ), rotavirus and papova-like virus. Parvovirus-like particles alone were detected in 14 diarrhoeal and 50 normal faeces, astrovirus-like particles in 3 normal faeces, "round" viruses in 4 normal faeces, coronavirus in 2 diarrhoeal and 5 normal faeces, CVLP in one diarrhoeal and one normal faeces, rotavirus in 2 normal faeces, papova-like virus in one normal faeces, both parvovirus-like particles and coronavirus in 2 diarrhoeal and 2 normal faeces, parvovirus-like particles and rotavirus in one normal faeces and parvovirus-like and papova-like virus in one normal faeces. The significance of these findings in canine and human disease is discussed.     

Isolation of coronaviruses from neonatal calf diarrhoea in Great Britain and Denmark

Coronaviruses were isolated from neonatal calves with diarrhoea in Great Britain and Denmark. They were serially passed in gnotobiotic calves which developed acute diarrhoea. Pathological lesions were found in the small and large intestines. Coronaviruses were demonstrated by electron microscopic examination of the faeces and intestinal contents, immunofluorescent staining of sections of small and large intestine and by isolation in tracheal organ cultures. In early passages of the British coronavirus, particles of about 30 nm in diameter were observed in the faeces by electron microscopy. These particles were removed from the coronavirus preparations by cross-protection experiments in calves. The coronaviruses were morphologically and antigenically similar to the bovine coronavirus isolated in the United States and the British virus was adapted to replicate in calf kidney cell cultures.     

从猝死症黄牛分离出冠状病毒

应用电镜观察，在吉林省猝死黄牛的心，肝，脾和肾中发现典型的冠状病毒颗粒，其中心，肝样品中的冠状病毒较多。用牛胎肾传代细胞（ＭＤＢＫ）分离培养该病毒获得成功，并进行了电镜负染跟踪观察和电镜超薄切片观察。该病毒呈圆形和椭圆形为主的多形性，直径７０～１６０ｎｍ有囊膜，囊膜表面有突起，突起呈间隙较宽的顶端膨大成球形的２０ｎｍ长的柄状结构，即呈鼓锤样结构。     

654—2对山羊内毒素性病理损害的保护效应

采用病理学和电镜技术观察了山莨菪碱（６５４－２）对内毒素（ＥＴ）诱导山羊病理损伤的影响。结果表明：ＥＴ处理组各组织器官充血、淤血,血管内有中性粒细胞和血小板聚集,并见透明血栓形成,血管内皮细胞肿胀,管壁透明变性、肝、肾等实质器官退行性变化,具有明显的超微结构变化。６５４－２能有效地缓解或减ＥＴ诱导的病理损伤。     

Lymphadenopathy in dogs associated with aluminosilicate

Granulomatous lymphadenopathy, associated with the presence of needle-like refractile particles, was recognised in two dogs. The material was detected in macrophages, either free within the cytoplasm or in membrane-bound lysosomes. By mineral analysis under direct vision in an electron microscope microanalyser (EMMA 4) the particles were found to contain aluminosilicate.     

鸡肾型传染性支气管炎病毒BK株的分离与鉴定

通过ＳＰＦ鸡胚传代培养、ＳＰＦ鸡胚气管环组织培养和鸡胚肾原代细胞培养,从一起有明显呼吸症状且又表现严重肾病变的鸡传染性支气管炎病例中分离到一株肾型传支病毒ＢＫ株。电镜观察表明,病毒粒子呈球状,直径９０～１００ｎｍ,纤突长１５ｎｍ,为典型冠状病毒。在ＳＰＦ鸡胚上盲传至５代,在ＴＯＣ上测其ＣＤ５０为１０－６．２。动物回归试验结果,试验鸡出现典型“花斑肾”,表明ＢＫ株具有较强嗜肾脏致病性。     

鸡传染性腺胃炎病原的分离与鉴定

自泰安地区的腺胃炎病鸡群中分离到TA毒株,在SPF鸡胚稳定传至第6代,病毒具有致鸡胚规律性死亡和典型致胚胎病变特征。TA株感染鸡胚尿囊液提纯的病毒经负染后电镜观察:病毒粒子大小为80 nm~160 nm,有囊膜,表面有纤突,呈典型冠状病毒形态。病毒无直接血凝性,但经10 g/L胰酶处理后能凝集鸡红细胞。对病毒进行了ELD50测定、对NDV的干扰试验和致病性试验。结果表明,病毒均呈现冠状病毒的特性和与自然病例相似的病理症状。     

鸭坦布苏病毒自然感染病例的诊断与病理学观察

为探究鸭坦布苏病毒(DTMUV)在贵州省鸭群的流行,对疑似DTMUV感染肉种鸭进行剖检,观察发病鸭内脏器官病变。取病鸭内脏组织进行RT-PCR检测确诊为DTMUV感染,将病料处理后接种BHK-21细胞,进行RT-PCR鉴定和透射电镜观察,确定分离到DTMUV,并将病料组织制作病理切片。结果显示,接种BHK-21出现明显CPE,电镜观察见病毒颗粒。将分离毒株的E基因经PCR扩增后测序,分析E基因氨基酸序列的同源性,发现所分离毒株与中国北京鸭源参考株同源性最高达99.39%,而与我国早期分离参考株(FX2010)同源性较低,将病料制作切片并进行HE染色,可见脑组织非化脓性脑炎,脾脏淋巴细胞减少,肝细胞变性坏死。本研究成功在贵州省分离到一株DTMUV,对其遗传进化进行分析,为探究贵州省养鸭地区DTMUV的感染和致病提供参考。