一株猫疱疹病毒Ⅰ型（FHV-1）的分离鉴定及攻毒模型建立

从一只临床表现为流脓涕、打喷嚏的猫身上分离出一株猫疱疹病毒Ⅰ型(Feline herpesvirus typeⅠ,FHV-1),使用F81细胞进行分离,并通过PCR、透射电镜和糖蛋白D(gD)基因序列分析来鉴定病毒。通过PCR方法从猫的鼻分泌物中扩增出1125 bp的gD基因。序列分析表明,该分离株与从GenBank获得的其他FHV-1毒株属于同一家族。随后在接种猫鼻分泌物的F81细胞的培养上清中观察到有包膜和直径约120 nm左右的疱疹样病毒颗粒。将分离获得的FHV-1病毒感染2～3月龄蓝猫,成功建立动物实验模型,感染后的猫出现脓性的眼鼻分泌物,打喷嚏等临床症状,且一周内持续排毒。该分离株为FHV-1的病原学研究提供了重要的数据,也为猫鼻气管炎疫苗候选株的筛选和疫苗的研发奠定了基础。     

A real‐time PCR for the detection of hepatopancreatic parvovirus (HPV) of penaeid shrimp

Hepatopancreatic parvovirus (HPV) causes a common shrimp disease that occurs in many shrimp farming regions, especially in the Indo Pacific, and infects most of the cultured penaeid species. There are seven geographic HPV isolates known, so a method to detect different HPV types is needed. We developed a sensitive and generic real‐time PCR assay for the detection of HPV. A pair of primers and TaqMan probe based on an HPV sequence obtained from samples of Fenneropenaeus chinensis from Korea were selected, and they were used to amplify a 92 bp DNA fragment. This real‐time PCR was found to be specific to HPV and did not react with other shrimp viruses. A plasmid (pHPV‐2) containing the target HPV sequence was constructed and used for determination of the sensitivity of this assay. The assay could detect a single copy of plasmid DNA, and it was used successfully in finding HPV in shrimp samples from the China‐Yellow Sea region, Taiwan, Korea, Thailand, Madagascar, New Caledonia and Tanzania.     

Aspergillus flavus keratomycosis in a cat treated with topical 1% voriconazole solution

An 8-year-old male castrated Domestic Short-haired cat was examined for a 1-week history of blepharospasm and mucoid ocular discharge OS. Examination revealed ulcerative keratitis with stromal loss, stromal infiltrate, corneal edema, perilimbal vascularization and miosis. Cytology of the cornea revealed multiple dichotomously branching, septate fungal hyphae and severe, predominantly neutrophilic inflammation. PCR of the cytology samples confirmed the presence of Aspergillus flavus while fungal and bacterial cultures were negative. Treatment with topical 1% voriconazole solution was successful in resolving the keratomycosis.     

A retrospective analysis of radiation therapy for the treatment of feline vaccine‐associated sarcoma*

We retrospectively evaluated predictive prognostic factors in 73 cats with vaccine‐associated sarcoma given postsurgical curative (n = 46, most with clean margins) or coarse fractionated radiotherapy (n = 27, most with either macroscopic disease or dirty margins). The former animals displayed a median survival of 43 months and a median progression free interval (PFI) of 37 months, the latter reached a median survival of 24 months and a median PFI of 10 months. In cats undergoing coarse fractionated therapy, factors predictive of a better outcome included lack of visible mass (n = 10) as opposed to macroscopic disease (n = 17, survival: 30 versus 7 months, P = 0.025; PFI: 20 versus 4 months, P = 0.01), adjuvant chemotherapy for gross disease (n = 5/17, survival: 29 versus 5 months, P = 0.04) and a smaller number of surgeries preceding radiation therapy (coeff = 0.41, P = 0.03). The Ki67 index was not predictive for survival. We concluded that postsurgical curative and coarse fractionated radiotherapy are effective legitimate options for managing vaccine‐associated sarcomas.     

Incidence and risk factors for development of malignant neoplasia after feline renal transplantation and cyclosporine-based immunosuppression

The objective of this retrospective study is to report and analyse the incidence of and risk factors for post-transplant malignant neoplasia (PTMN) in feline renal transplant recipients (cases, n  = 45) and compare incidence to a population of cats that did not receive a transplant (controls, n  = 79). Information from the medical records of cases regarding signalment, blood work and concomitant disease, post operative cyclosporine concentrations, survival time (ST), and whether PTMN developed, the type of PTMN, time to occurrence (TTO), and ST after diagnosis was gathered. PTMN occurred in 11 of 45 (24%) cases, of which, four were lymphoma. Median TTO of all PTMN was 1020 days. Median TTO of lymphoma was 454 days. Median ST after diagnosis of PTMN was 15 days. No risk factors were identified. Compared with control cats, cases had more than six times higher odds of developing PTMN compared with control cats ( P  < 0.01).     

Validation of data collected in the Danish Veterinary Cancer Registry

The Danish Veterinary Cancer Registry (DVCR) was established in 2005 and prospectively collects information about neoplasms in Danish dogs and cats.
The present study evaluated the agreement between veterinary practice records and electronic submissions made to the DVCR from May 2005 through June 2008. The variables compared were gender, breed, diagnosis, date of birth, death and diagnosis, localization and biological behaviour of the neoplasms.
Agreement of 95% between DVCR and the original data were considered acceptable with regard to use of data in studies. Recorded proportions of agreement were: (1) breed: 97.4%; (2) diagnosis: 95.6%; (3) location: 95.6%; (4) biological behaviour: 93.0%; (5) gender: 92.5%; (6) date of death: 91.5%; (7) date of diagnosis: 80.1%; (8) date of birth: 76.7%.
All variables except date of death, birth, and diagnosis met the required 95% agreement limit. Data on dates of birth, death and diagnosis were considered less suitable for further studies.     

Topical tacrolimus (FK506) for the treatment of feline idiopathic facial dermatitis

A 3-year-old, neutered male Persian cat with chronic ulcerative facial dermatitis was diagnosed with feline idiopathic facial dermatitis based on signalment, clinical history and diagnostic test results, including dermatohistopathological evaluation. Initial treatment started with 4 weeks of oral antifungal/antibiotic medication for severe secondary infectious dermatitis of Malassezia and bacteria. As the lesions gradually improved, the oral medication was withdrawn, leaving only 0.1% topical FK506 (tacrolimus) ointment for the remaining lesions. Topical treatment was administered just in case any new lesions developed. The patient has been managed effectively with topical tacrolimus and no side-effects were observed during treatment. Feline idiopathic facial dermatitis is known as a difficult dermatosis to manage successfully, but our experience suggests that it may respond to topical tacrolimus.     

猪细小病毒在PK15细胞中增殖规律的研究

猪细小病毒毒株在不同细胞系上的生长情况不尽相同,如不能掌握其规律,将难以生产出优质的疫苗。对PPV（长春株）在PK15和ST细胞上生长情况进行了研究,在PK15细胞系中重点比较了不同的病毒接种方法、病毒接种量和病毒收获时间等因素对病毒产量的影响。研究结果显示：本毒株在PK15和ST两种细胞系上的病变特征不同,但生长规模相似。在PK15细胞上按2％接种量、分步接种、80h时收获病毒,最终得到的病毒滴度（TCID50）最高。提示这是一种比较好的生产方案。然而,进一步的大规模培养试验是有必要的。     

猪细小病毒SYBR Green Ⅰ模式实时定量PCR检测方法的建立

根据GenBank猪细小病毒(PPV)的NS1基因序列,设计一对特异性引物,采用SYBR Green Ⅰ随机结合渗入法,建立实时定量PCR检测方法,构建了检测PPV的标准DNA模版,循环阈值(Ct)与标准质粒DNA模板在7.8×101～7.8×108拷贝/μl浓度范围内呈良好的线性关系,相关系数为0.997。该方法用于猪细小病毒的检测具有很高的特异性,其敏感性与常规PCR相比可以提高100倍,可以用于猪细小病毒的快速检测。     

IN VITRO EVALUATION OF FELINE LEUKOCYTES RADIOLABELED IN WHOLE BLOOD WITH 99MTC STANNOUS COLLOID

Technetium-99m stannous colloid (^99mTcSnC) has been used to radiolabel human leukocytes to investigate various inflammatory disorders. We investigated the in vitro behavior of feline leukocytes labeled in whole blood with ^99mTcSnC. Heparinized blood samples were collected from healthy cats and divided into control and test aliquots. The latter were labeled with ^99mTcSnC using a standard procedure. Leukocyte viability was determined for each sample using a trypan blue exclusion test. Labeling efficiency was determined for test aliquots. Test aliquots were layered onto Histopaque-1077^® and centrifuged before measurement of radioactivity of the blood components. Leukocytes from radiolabeled and control samples were washed and incubated with opsonized zymosan particles to allow assessment of phagocytic function. Aliquots were taken from radiolabeled feline leukocyte samples at 1, 3, 4, and 7 h postlabelling. After centrifugation of each aliquot, radioactivity of the supernatant and pellet was measured and the labeling retention determined. Leukocyte viability in both radiolabeled and control samples was >98%. The labeling efficiency was 95.2±0.14%. The distribution of radioactivity in feline blood was found to be 3.4±0.18% in plasma, 39.0±0.37% in erythrocytes, and 57.6±0.38% in leukocytes. Labeled feline leukocytes had phagocytic activity of 90.9±0.18% (control 91.3±0.15%). The radiolabeled leukocytes retained 93.4±0.19% of the radioactivity up to 7 h postlabeling. ^99mTcSnC efficiently labeled feline leukocytes with no effect on viability and minimal effect on phagocytic function. The percentage retention of radioactivity by the leukocytes was still high at 7 h postlabeling.