山东省鸡新城疫病毒弱毒株的初步分离

某鸡场饲养的罗曼产蛋鸡群爆发了典型的新城疫,采集病料接种１１日龄ＳＰＦ鸡胚进行病毒分离。经致鸡胚病变,回归试验、血凝和血凝抑制试验结果表明,所分离到的病毒系鸡新城疫病毒弱毒株,定名为ＮＤＶ－Ｓ１。     

新城疫病毒强毒株的分离与鉴定

从发病率为１００％、病死率为９７％的鸡群中分离到１株病毒。该病毒凝集鸡红细胞的作用可被新城疫标准阳性血清所抑制,证明所分离的毒株为新城疫病毒。鸡胚半数致死量、最小致死量致死鸡胚的平均时间、１日龄雏鸡脑内致死指数、６周龄雏鸡静脉致死指数、血凝解脱及血凝素热稳定性等试验表明,分离毒为新城疫病毒强毒株,其毒力比标准强毒Ｆ４８Ｅ８株还强。血凝抑制试验和中和试验表明,分离毒的血凝性与Ｆ４８Ｅ８相同,但中和特性与Ｆ４８Ｅ８有较大差异     

猪源新城疫病毒的分离鉴定

从发生呼吸道症状的病猪鼻腔中分离到1株病毒，经电镜观察、血凝和血凝抑制试验，确定为新城疫病毒。经对鸡胚平均致死时间、鸡胚半数致死量、对鸭胚和鸡、鸭的致死性测定及荧光PCR检测，证明分离毒为温和型毒株。     

犬细小病毒四川株的分离与鉴定

采集5份患出血性肠炎犬的粪便,通过猫肾细胞(F81)传代培养后,成功分离到2株病毒。对2株病毒进行病毒形态学、理化学、血凝试验、PCR鉴定与分子生物学系统鉴定后,证实分离株是犬细小病毒,通过VP2结构蛋白测序分析表明,新分离到的2株细小病毒抗原型分别属于CPV-2a和CPV-2b。     

腺胃型鸡传染性支气管炎病毒的初步分离与鉴定

为了获得更多的腺胃型鸡传染性支气管炎病毒分离株并研究其生物学特性,试验从黑龙江省某鸡场发生的以腺胃肿大为特征的病死鸡中分离到1株具有冠状病毒特性的病毒,通过鸡胚传代、血凝性测定、电镜观察、RT-PCR检测和动物回归试验对其进行了初步鉴定。结果表明:该病毒分离株在SPF鸡胚上传至第5代开始出现死亡或典型的侏儒胚;病毒不能直接凝集鸡红细胞,经1%胰酶处理后表现出血凝活性,血凝效价达28;透射电镜下可见有囊膜和纤突,直径为80～120nm的病毒粒子,具有冠状病毒的典型形态特点;分离株病毒第5代尿囊液经N基因特异性RT-PCR获得大小约438bp的目的片段;分离株病毒第5代尿囊液接种于14日龄SPF雏鸡,复制出与临床中相似的病理变化,同时从肿大的腺胃中能重新分离到该病毒。初步确定所分离病毒为腺胃型鸡传染性支气管炎病毒。     

果子狸源细小病毒分离鉴定及VP2基因遗传进化分析

【目的】探究果子狸源细小病毒流行特点及其VP2基因遗传变异情况,为细小病毒防治提供理论依据和技术支撑。【方法】采用猫肾细胞(CRFK细胞)对患有肠炎的果子狸肠道组织样品进行病毒分离,通过血凝试验、免疫荧光试验鉴定分离的病毒,并对病毒VP2基因进行PCR扩增和遗传进化分析。【结果】分离到的毒株在CRFK细胞中培养出现细胞脱落、崩解和破碎等典型细小病毒细胞病变效应(CPE);血凝试验结果显示,此病毒可凝集猪红细胞,血凝效价为1∶512;免疫荧光试验结果显示,在接种分离毒株的CRFK细胞内可观察到亮绿色荧光,而对照细胞没有荧光。将分离毒株命名为MPCPV-SX。VP2基因进化树分析发现,其与犬细小病毒处于同一分支,位于CPV-2和CPV-2a型之间,同时VP2氨基酸的87和101位显示为CPV-2型,而300和305位氨基酸则与CPV-2a型一致。【结论】本研究成功从果子狸肠道组织样品中分离出1株细小病毒MPCPV-SX,其VP2基因是介于CPV-2和CPV-2a之间的中间型,表明细小病毒通过果子狸等野生动物跨越宿主流行传播,可能在细小病毒遗传进化方面起到一定的过渡作用。     

免疫鸡发生肾型传染性支气管炎

湖北某蛋鸡场曾用ＭＡ５（英特威）和肾型传支（华中农大）免疫过的鸡群发生了以肾脏肿大、尿酸盐沉积为主要特征的传染病 ,从中分离到一株肾型传染性支气管炎病毒株 ,命名为ＨＰ２。对其培养特性、血凝性、对新城疫病毒干扰性、动物回归等方面进行了初步研究。结果表明 :该毒株能很快适应于９～１１日龄鸡胚 ,并引起鸡胚特征性病变 ;该病毒对鸡红细胞无凝集作用 ,经１ %胰酶处理后表现血凝性 ;病毒能抑制鸡新城疫病毒ＬａＳｏｔａ株在９～１１日龄鸡胚上增殖 ;分离毒株在未免疫鸡群中有致病性 ;并应用自家组织灭活苗有效地控制了本病的流行…     

犬细小病毒四川株的分离与鉴定

为更好地了解我国细小病毒流行情况,笔者从四川农大动物医院采集5份患出血性肠炎犬的粪便,通过猫肾细胞(F81)传代培养后,成功分离到两株犬细小病毒。对两株病毒进行病毒形态学、理化学、血凝试验、PCR鉴定与分子生物学系统鉴定后,证实分离株是犬细小病毒,通过VP2结构蛋白测序分析表明,新分离到的两株细小病毒抗原型分别属于CPV-2a和CPV-2b。     

禽肺病毒的分离鉴定

禽肺病毒（ＡＰＶ）于１９７８年首次发现于南非,可引起火鸡鼻气管炎和鸡肿头综合症,至今未见其在中国存在的报道。我们从黑龙江省某肉种鸡场患肿头综合症的鸡中分离到了１株病毒。经鉴定,该病毒分离株缺乏血凝活性,具有典型的副粘病毒形态特征,可诱导特异性抗ＡＰＶ抗体的产生。因此可认定该病毒为１个ＡＰＶ分离株（命名为ＡＰＶ／Ｃｈｉｃｋｅｎ／Ｃｈｉｎａ／１／９８）。     

一株鸡肾型传染性支气管炎病毒的分离与鉴定

从山东某鸡场疑似肾型传染性支气管炎病死鸡的肾脏中分离到一株病毒,经SPF鸡胚连续传代、血凝试验、鸡新城疫干扰试验、鸡胚矮小化试验、RT—PCR鉴定和动物回归试验,初步确定该病毒为肾型传染性支气管炎病毒,并命名为SD1105株。     

Isolation and characteristics of an equine reovirus type 3 and an antibody prevalence survey to reoviruses in horses located in New York State

Reoviruses have been isolated from a number of species including human, bovine, feline, canine and equine. In most species they seem to produce mild to inapparent disease. We have isolated a reovirus type 3 from a foal with diarrhea. The virus designated the Ralph strain has been propagated in both the MA-104 and A-72 cell lines. The strain produced cytoplasmic inclusion bodies in these cell cultures. Tissue-cultured virus fixed complement in the presence of reovirus antibodies, but failed to do so in the presence of rotavirus antiserum. By electron microscopy the viral particle measured +/- 65 nm. The virus hemagglutinated pig erythrocytes, but not human O, human A, calf, cow, chicken or guinea pig erythrocytes. In the hemagglutination test there was complete reciprocal crossing between the Ralph strain and the NIH reovirus type 3, but there was no crossing with the NIH reovirus types 1 and 2. A limited serological survey was completed on serum samples from New York State horses collected in 1976-1977 and 1981 using the hemagglutination-inhibition test. The percentage with antibodies to reovirus types 1, 2 and 3 for 1976-1977 was 24.5, 42.2 and 3.9% and in 1981, 8.8, 9.8 and 3.9%, respectively.     

Virological and molecular characterization of a mammalian orthoreovirus type 3 strain isolated from a dog in Italy

A mammalian orthoreovirus (MRV) strain was isolated from a pup with fatal diarrhea, which had a concurrent infection by canine parvovirus type 2. The reovirus isolate showed an atypical hemagglutination pattern and a retarded electrophoretic mobility of the S1 segment, which is characteristic of MRV type 3 (MRV-3). Assignment of the isolated virus to MRV-3 was confirmed by type-specific RT-PCR assays, targeting the S1 gene, and by subsequent sequence analysis of the PCR product. By phylogeny based on the S1 gene of several MRVs, the isolate fell into lineage E, along with the murine strain T3C9/61 and the bovine strains T3C18/61 and T3C31/59. Conversely, L1 sequences were found to segregate regardless of the viral type. A total of 110 fecal samples, 56 nasal and 31 ocular swabs from dogs with diarrhea or nasal/ocular discharge were tested by a nested-PCR assay specific for reoviruses, and no sample was found to contain MRV RNA, a finding that is apparently in contrast with the seroprevalence (25.77%) observed in dogs.     

犬呼肠病毒3型在BHK-21细胞上增殖的研究

采用酶标ＳＰＡ法进行了犬呼肠３ 型病毒在ＢＨＫ２１ 细胞上定位的研究。结果表明：接毒后６ ｈ 即可在胞浆见到絮状阳性反应,２４ ｈ 后呈强阳性着染;Ｈ．Ｅ．染色在胞浆内可见强嗜酸性着染或嗜酸性包涵体,提示可能是病毒感染引起的特征性病变,感染后细胞病变发展较快,６０ ｈ 细胞大部脱落;病毒血凝滴度结果也证实了细胞病变与病毒增殖有关。     

Oncolytic reovirus synergizes with chemotherapeutic agents to promote cell death in canine mammary gland tumor

The oncolytic effects of reovirus in various cancers have been proven in many clinical trials in human medicine. Oncolytic virotherapy using reovirus for canine cancers is being developed in our laboratory. The objective of this study was to examine the synergistic anti-cancer effects of a combination of reovirus and low doses of various chemotherapeutic agents on mammary gland tumors (MGTs) in dogs. The first part of this study demonstrated the efficacy of reovirus in canine MGTs in vitro and in vivo. Reovirus alone exerted significant cell death by means of caspase-dependent apoptosis in canine MGT cell lines. A single injection of reovirus impeded growth of canine MGT tumors in xenografted mice, but was insufficient to induce complete tumor regression. The second part of this study highlighted the anti-tumor effects of reovirus in combination with low doses of paclitaxel, carboplatin, gemcitabine, or toceranib. Enhanced synergistic activity was observed in the MGT cell line treated concomitantly with reovirus and in all the chemotherapeutic agents except toceranib. In addition, combining reovirus with paclitaxel or gemcitabine at half dosage of half maximal inhibitory concentration (IC₅₀) enhanced cytotoxicity by activating caspase 3. Our data suggest that the combination of reovirus and low dose chemotherapeutic agents provides an attractive option in canine cancer therapy.     

The oncolytic effects of reovirus in canine solid tumor cell lines

Oncolytic virotherapy is a new strategy for cancer treatment for humans and dogs. Reovirus has been proven to be a potent oncolytic virus in human medicine. Our laboratory has previously reported that canine mast cell tumor and canine lymphoma were susceptible to reovirus. In this study, canine solid tumor cell lines (mammary gland tumor, osteosarcoma and malignant melanoma) were tested to determine their susceptibility towards reovirus. We demonstrated that reovirus induces more than 50% cell death in three canine mammary gland tumors and one canine malignant melanoma cell line. The reovirus-induced cell death occurred via the activation of caspase 3. Ras activation has been shown to be one of the important mechanisms of reovirus-susceptibility in human cancers. However, Ras activation was not related to the reovirus-susceptibility in canine solid tumor cell lines, which was similar to reports in canine mast cell tumor and canine lymphoma. The results of this study highly suggest that canine mammary gland tumor and canine malignant melanoma are also potential candidates for reovirus therapy in veterinary oncology.     

Anti‐tumour activity of oncolytic reovirus against canine histiocytic sarcoma cells

Canine histiocytic sarcoma is an aggressive, fatal neoplastic disease with a poor prognosis. Lomustine is generally accepted as the first‐line systemic therapy, although this compound does not provide complete regression. Therefore, research into a novel approach against canine histiocytic sarcoma is needed. However, anti‐tumour effects of oncolytic therapy using reovirus against histiocytic sarcoma are unknown. Here, we showed that reovirus has oncolytic activity in canine histiocytic sarcoma cell lines in vitro and in vivo. We found that reovirus can replicate and induce caspase‐dependent apoptosis in canine histiocytic sarcoma cell lines. A single intra‐tumoural injection of reovirus completely suppressed the growth of subcutaneously grafted tumours in NOD/SCID mice. Additionally, we demonstrated that susceptibility to reovirus‐induced cell death was attributable to the extent of expression of type I interferons induced by reovirus infection in vitro. In conclusion, oncolytic reovirus appears to be an effective treatment option for histiocytic sarcoma, and therefore warrants further investigation in early clinical trials.     

中国畜牧兽医2013年第40卷第2期犬流感(H3N2亚型)血清学调查

为了解广东地区犬流感(CI)流行情况,以期为犬流感的预防提供参考依据,本研究采用ELSIA方法及血凝抑制试验(HI),对广东省内6地市的1440份犬血清样品进行抗体检测。结果显示,1440份犬血清中犬流感抗体平均阳性率为7.92%(ELISA)和6.25%(HI)。其中广州、深圳和惠州地区的犬流感抗体阳性率较高。1～3岁犬的犬流感抗体阳性率高于其他年龄段犬,雄性犬犬流感抗体阳性率高于雌性。结果表明,H3N2亚型犬流感病毒在广东局部地区流行。鉴于当前在部分地区CIV阳性率较高,结合CIV在犬群中迅速传播的流行特点,要避免CIV暴发流行及对人类的健康造成威胁,对于CIV的防制应结合当地实际情况,制定切实有效的防制措施。     

Serotype, hemolysin production, and adherence characteristics of strains of Escherichia coli causing urinary tract infection in dogs.

Virulence factors were studied in 82 strains of Escherichia coli isolated from the urine of dogs with urinary tract infections. The most frequently expressed O antigens were 2, 4, 6, 25, and 22/83. Most strains were K nontypeable. Mannose-sensitive hemagglutination (MSH) with canine erythrocytes was observed in 71 strains and mannose-resistant hemagglutination (MRH) was observed in 32 strains. Strains that caused MSH of erythrocytes from dogs also caused MSH of erythrocytes from guinea pigs. Most strains that caused MRH of human A1P1 erythrocytes also reacted with erythrocytes of dogs. Of 22 strains (27%) that agglutinated human A1P1 erythrocytes, but not A1p erythrocytes, 17 (77%) had specificity for globo A, but did not react with the galactose alpha 1----4galactose beta disaccharide receptor. The remaining 5 strains and 2 others that simultaneously expressed an X adhesin agglutinated galactose alpha 1----4galactose beta-coated latex beads. Bacterial adherence to canine uroepithelial cells from the bladder was most often observed in strains expressing MSH, less often observed in strains expressing MRH, and least often observed in strains that failed to induce hemagglutination. Adherence of MSH strains to canine uroepithelial cells was inhibited by alpha-methyl-D-mannoside. As a group, MRH strains expressing globo-A- and galactose alpha 1----4galactose beta-specific adhesins did not have strong adherence. Strains of E coli isolated from dogs with urinary tract infections most commonly expressed type-1 fimbriae, and the main mechanism of in vitro adherence to canine uroepithelial cells involved a mannose-sensitive mechanism. Overrepresentation of globo-A-specific adhesins did not appear to be related to adherence of canine uroepithelial cells.     

一株貉源犬瘟热病毒的分离与鉴定

从临床疑似犬瘟热的病貉组织中分离到1株病毒,经形态特征、血凝试验、中和试验、间接免疫荧光试验和分子生物学鉴定,该分离病毒为犬瘟热病毒。本研究为研究犬瘟热病毒貉株分子生物学特性、致病机理及构建基因工程疫苗奠定了基础。     

犬传染性喉气管炎病毒的分离及鉴定

用犬肾细胞 (MDCK)从疑似犬传染性喉气管炎病毒感染犬的急性期血清中分离到 1株病毒 ,经血凝试验、免疫电镜观察、中和试验、免疫荧光抗体试验、细胞培养特征观察和回归动物试验等鉴定 ,证明所分离的病毒为犬传染性喉气管炎病毒 ,命名为 BJ- JB- 3。