Isolation of feline calicivirus and feline herpesvirus from domestic cats 1980 to 1989

Isolation rates of feline herpesvirus (FHV) and feline calicivirus (FCV) from oropharyngeal swabs, taken from 6866 cats in 1980 to 1989 were studied retrospectively. FCV was isolated from 1364 (19.9 per cent) and FHV from 285 (4.2 per cent). The ratio of FCV:FHV isolations varied from 1.3:1 to 15:1 in individual years with an overall ratio of 4.8:1. Isolation of both viruses was fairly uniform for each year and there was no breed or sex disposition to either virus. Of 872 cats shedding FCV and 213 cats shedding FHV, of known age, 447 (51.3 per cent) with FCV and 140 (65.7 per cent) with FHV were under one year old, compared to only 35.3 per cent of the whole population sampled. For the years 1985 to 1989, more information was obtained about the cases. Of 4626 cats tested, 1180 (25.5 per cent) had acute upper respiratory tract disease (URTD) of which 348 (29.5 per cent) were shedding FCV and 162 (13.7 per cent) FHV. A further 597 had chronic URTD and of these, 102 (17.1 per cent) were shedding FCV and 18 (3 per cent) FHV. In 120 cases of suspected vaccine reaction/breakdown, FCV was isolated from 34 (28.3 per cent) and FHV from only two (1.7 per cent). FHV was not isolated from any of 412 cases presenting with chronic gingivitis/stomatitis alone; 181 (43.9 per cent) were shedding FCV and when cats with other signs in addition to chronic gingivitis were included, this proportion increased to 70.4 per cent.(ABSTRACT TRUNCATED AT 250 WORDS)     

Efficacy of an inactivated feline panleucopenia virus vaccine against a canine parvovirus isolated from a domestic cat

Canine parvovirus type 2a (CPV-2a) and type 2b (CPV-2b) have recently been isolated from cats throughout the world, and CPV-2b strain FP84 has been reported to be virulent in domestic cats. Although live feline panleucopenia virus (FPLV) vaccines protect domestic cats from CPV infection, the efficacy of inactivated FPLV vaccines has not been established. In this study, two domestic cats were vaccinated with a commercial inactivated FPLV vaccine and challenged with CPV-2b strain FP84 isolated from a domestic cat. The cats were protected against CPV-2b strain FP84 infection and their clinical signs were suppressed, although the two unvaccinated cats showed the typical clinical signs of parvovirus infection.     

Molecular virology of feline calicivirus.

Caliciviridae are small, nonenveloped, positive-stranded RNA viruses. Much of our understanding of the molecular biology of the caliciviruses has come from the study of the naturally occurring animal caliciviruses. In particular, many studies have focused on the molecular virology of feline calicivirus (FCV), which reflects its importance as a natural pathogen of cats. FCVs demonstrate a remarkable capacity for high genetic, antigenic, and clinical diversity; "outbreak" vaccine resistant strains occur frequently. This article updates the reader on the current status of clinical behavior and pathogenesis of FCV.     

An isolated epizootic of hemorrhagic-like fever in cats caused by a novel and highly virulent strain of feline calicivirus

An isolated epizootic of a highly fatal feline calicivirus (FCV) infection, manifested in its severest form by a systemic hemorrhagic-like fever, occurred over a 1-month period among six cats owned by two different employees and a client of a private veterinary practice. The infection may have started with an unowned shelter kitten that was hospitalized during this same period for a severe atypical upper respiratory infection. The causative agent was isolated from blood and nasal swabs from two cats; the electron microscopic appearance was typical for FCV and capsid gene sequencing showed it to be genetically similar to other less pathogenic field strains. An identical disease syndrome was recreated in laboratory cats through oral inoculation with tissue culture grown virus. During the course of transmission studies in experimental cats, the agent was inadvertently spread by caretakers to an adjoining room containing a group of four normal adult cats. One of the four older cats was found dead and a second was moribund within 48-72h in spite of symptomatic treatment; lesions in these animals were similar to those of the field cats but with the added feature of severe pancreatitis. The mortality in field cats, deliberately infected laboratory cats, and inadvertently infected laboratory cats ranged from 33-50%. This new isolate of calicivirus, named FCV-Ari, was neutralized at negligible to low titer by antiserum against the universal FCV-F9 vaccine strain. Cats orally immunized with FCV-F9, and then challenge-exposed shortly thereafter with FCV-Ari, developed a milder self-limiting form of disease, indicating partial protection. However, all of the field cats, including the three that died, had been previously immunized with parenteral FCV-F9 vaccine. FCV-Ari caused a disease that was reminiscent of Rabbit Hemorrhagic Disease, a highly fatal calicivirus infection of older rabbits.     

Characterization of a wild-type strain of Francisella tularensis isolated from a cat.

Francisella tularensis type A is the primary cause of tularemia in animals and humans in North America. The majority of research on F. tularensis has been done with the attenuated live vaccine strain (LVS), which is a type B, but very few wild-type F. tularensis strains have been characterized. A gram-negative coccobacillus that was isolated in pure culture from the lungs of a cat that died after being lost for 5 days was received for identification at the Virginia-Maryland Regional College of Veterinary Medicine Teaching hospital. The isolate (strain TI0902) was not identified (or was misidentified) by commercial identification systems; however, it was identified as F. tularensis subspecies tularensis (type A) by sequencing a portion of the 16S ribosomal RNA gene. Furthermore, repetitive extragenic palindromic sequences-polymerase chain reaction amplified a 4-kb DNA fragment from TI0902 that was characteristic of F. tularensis type A but not type B. The electrophoretic profile of the lipopolysaccharide of strain TI0902 was identical to that of the LVS by Western blotting with antiserum to LVS. The protein-enriched outer membrane of strain TI0902 contained 6-8 proteins, which were similar in molecular size to those from the LVS. Electron microscopy of negatively stained and alcian blue-stained LVS and TI0902 cells showed that both strains were coccobacillary in shape and may be encapsulated. However, after mouse challenge, the TI0902 strain was clearly more virulent than the LVS strain. Results of this study indicate that the genotype and phenotype of wild-type F. tularensis type A strain TI0902 is similar, but not identical, to that of the LVS strain. Further studies will help determine whether pathogenesis and host-pathogen interactions are also similar between the 2 strains.     

Immunization against feline calicivirus infection.

Forty-three cats (experiments 1 and 2) were vaccinated (2 doses, 27 and 30 days between doses) with the F-9 strain of feline calicivirus by the intramuscular route. There was no untoward response in any of the cats to the administration of the vaccinal virus nor was there spread of the virus from 20 vaccinated cats to nonvaccinated cats held in contact during the next 6 months (experiment 2). The vaccinated cats developed serum-neutralizing antibodies that were increased further after the 2nd vaccination. The level of serum-neutralizing antibodies was related to the quantity of vaccinal virus administered. Twenty-three cats vaccinated with the F-9 strain were protected to a significant degree when challenge exposed to virulent calicivirus strain FPV-255 (experiment 1).     

Cessation of feline calicivirus shedding coincident with resolution of chronic gingivostomatitis in a cat

Feline calicivirus (FCV) shedding and oral bacterial flora were monitored over a period of 22 months in a case of feline gingivostomatitis (FGS). The cat was treated daily with 50 mg thalidomide capsules by mouth, and 200 mg lactoferrin powder was applied directly to the lesions. Clinical signs began to resolve after 11 months when, in addition to treatment, the diet had been changed to an additive-free cat food supplemented with antioxidant vitamins A, D3 and E. Resolution of clinical signs of FGS coincided with the cessation of FCV shedding, and this is the first report documenting such an association. Which part of the treatment, if any, contributed to the cure requires further investigation.     

The preparation of a polyvalent feline calicivirus antiserum.

A polyvalent antiserum capable of neutralizing 82 isolates of feline calicivirus made from cats in various parts of North America was produced by the sequential inoculation of SPF cats at three-week intervals with feline calicivirus strains F-9, 68-2024 and FS, followed by a final booster inoculation two weeks after the third inoculation with all three strains combined. Sera raised against the same strains but individually and then pooled failed to show such broad cross-neutralizing capacity. The polyvalent serum should prove useful for the confirmation of an isolation of feline calicivirus.     

The pathogenicity of canine parvovirus type-2b, FP84 strain isolated from a domestic cat, in domestic cats

Canine parvovirus type-2a (CPV-2a) and type-2b (CPV-2b) have recently been isolated from domestic cats. The pathogenicity of CPV-2b in domestic cats is still unclear. In this study, we performed infection tests to examine the pathogenicity of CPV-2b, FP84 strain, isolated from a domestic cat. The results demonstrated that the CPV strain FP84 is able to infect and replicate well in domestic cats. Two of the 3 cats used in the test died. They showed loss of appetite, diarrhea, leukopenia and dehydration. Since FP84 was found to be virulent to domestic cats, it is necessary to examine the efficacy of inactivated feline panleukopenia virus vaccines against CPV infection in domestic cats.     

Bilateral subcutaneous fibrosarcomas in a cat following feline parvo-, herpes- and calicivirus vaccination

A crossbred cat developed a subcutaneous fibrosarcoma on the left side of the thorax at the site of previous administration of a feline parvo-, herpes- and calicivirus vaccine. A few months later the cat developed a second mass on the right side of the thorax after a booster vaccine had been administered at this site. This unique case of bilateral fibrosarcomas in a cat shortly after vaccination with parvo-, herpes- and caliciviruses suggests an individual disposition for the development of vaccine-associated sarcomas and a possible triggering of this type of pathological response which could have precipitated the development of the second tumour. To the authors' knowledge, this is the first case of vaccine-induced fibrosarcomas occurring bilaterally after injection of a feline parvo-, herpes- and calicivirus containing vaccine at different sides of the thorax.     

Characterisation of canine parvovirus strains isolated from cats with feline panleukopenia

Unlike the original canine parvovirus type 2 (CPV-2), CPV-2 variants have gained the ability to replicate in vivo in cats but there is limited information on the disease patterns induced by these variants in the feline host. During 2008, two distinct cases of parvoviral infection were diagnosed in our laboratories. A CPV-2a variant was identified in a 3-month-old Persian kitten displaying clinical sign of feline panleukopenia (FPL) (acute gastroenteritis and marked leukopenia) and oral ulcerations, that died eight days after the onset of the disease. Two pups living in the same pet shop as the cat were found to shed a CPV-2a strain genetically identical to the feline virus and were likely the source of infection. Also, non-fatal infection by a CPV-2c strain occurred in a 2.5-month-old European shorthair kitten displaying non-haemorrhagic diarrhoea and normal white blood cell counts. By sequence analysis of the major capsid protein (VP2) gene, the feline CPV-2c strain showed 100% identity to a recent canine type-2c isolate. Both kittens had been administered multivalent vaccines against common feline pathogens including FPL virus. Whether and to which extent the FPL vaccines can protect cats adequately from the antigenic variants of CPV-2 should be assessed.     

Evaluation of a feline viral rhinotracheitis-feline calicivirus disease vaccine.

An attenuated respiratory disease vaccine against feline viral rhinotracheitis (FVR) and feline calicivirus (FCV) disease was evaluated for safety and efficacy in specific-pathogen-free cats. Twenty cats were vaccinated twice intramuscularly, with 28 days between vaccinations. Ten unvaccinated cats were used as contact controls. Adverse effects were not noticed after vaccination, and the vaccinal virus did not spread to contact controls. Arithmetical mean serum-neutralizing titers against vaccinal FCV strain F9 and challenge FCV strain 255 were 1:13 and 1:15 at 28 days after the 1st inoculation. These titers increased to 1:45 and 1:196 after the 2nd inoculation. After challenge exposure of vaccinated cats to virulent FCV 255 virus, mean titers increased to 1:129 and 1:865, respectively for F9 and 255 viruses. The F9 postchallenge mean titer for vaccinated cats was 21.5 times higher than that for the 8 contact controls that survived challenge exposure. The arithmetical mean serum neutralizing titer for FVR was low (1:2) after the 1st vaccination, but increased to 1:35 after the 2nd vaccination. Challenge exposure to virulent FVR virus resulted in a marked anamnestic immune response (mean titer of 1:207, compared with 1:12 for contact controls). In general, vaccinated cats remained alert and healthy after challenge exposure with FCV-255, whereas unvaccinated contact control cats developed definite signs of FCV disease, including central nervous system (CNS) depression (6 of 10) and dyspnea indicative of pneumonia (5 of 10). Two controls died of severe pneumonia. A mild fibrile response was detected in 28% of vaccinated cats, compared with a more severe febrile response in 78% of control cats. Some vaccinated cats developed minute lingual ulcers that did not appear to be detrimental to the health of the cat. After FVR challenge exposure, vaccinated cats were free of serious clinical signs. Five of 18 vaccinated cats had mild signs of FVR, including an occasional sneeze, low temperature, and mild serous lacrimation for 1 or 2 days. Contact controls developed definite clinical signs of FVR. The combined FVR-FCV vaccine appears to be safe and reasonably efficacious. Vaccination against FCV disease and FVR should be part of the routine feline immunization program.     

Update on feline calicivirus: new trends.

In addition to being important upper respiratory tract pathogens of cats, FCVs are increasingly reported as a cause of a highly contagious febrile hemorrhagic syndrome. Strains causing this syndrome are genetically different from the vaccine strain and other nonhemorrhagic FCV isolates. They apparently differ from one outbreak to another. The syndrome is characterized variably by fever; cutaneous edema and ulcerative dermatitis; upper respiratory tract signs; anorexia; occasionally icterus, vomiting, and diarrhea; and a mortality that approaches 50%. Adult cats tend to be more severely affected than kittens, and vaccination does not appear to have a significant protective effect. Rapid recognition of the disease through identification of clinical signs and appropriate testing, followed by strict institution of disinfection, isolation, and quarantine measures, are essential to prevent widespread mortality resulting from the infection.     

1株猫杯状病毒的分离鉴定及其ORF2序列的遗传演化分析

为了解猫杯状病毒形态特征及遗传演化情况,采用F81细胞从患病宠物猫的鼻拭子样品中分离获得1株猫杯状病毒(feline calicivirus,FCV),命名为SH1.经电镜观察,病毒粒子呈球形,无囊膜,符合FCV的形态特征.采用RT-PCR方法扩增了该毒株的全基因组,并进行了序列测定和衣壳蛋白基因(ORF2)序列的分析...     

Carrier-state infection of feline T-lymphoblastoid cells with feline calicivirus

The susceptibility of feline T lymphocytes to feline calicivirus (FCV) in vitro was investigated using feline T-lymphoblastoid cell lines, namely MYA-1 and FL74 cells. The virus titers of supernatants in FCV-infected MYA-1 and FL74 cell cultures increased rapidly, and FCV antigens were also detected in the FCV-infected cells. There were slight differences in the molecular weights of capsid proteins expressed in FCV-infected MYA-1, FL74 and Crandell feline kidney cells. MYA-1 and FL74 cells were productively and persistently infected with FCV, and FCV antigens were observed in the FCV-infected cells for more than one month. At 3 months post infection, FCV-infected FL74 cells that stopped producing infectious FCV could be reinfected with FCV. However, no cytopathic effects were observed.     

Sites of persistence of feline calicivirus

Various tissues were collected from eight cats persistently infected with feline calicivirus (FCV) strain 255 to determine the sites of viral persistence. Tissues were tested by virus isolation and an immunohistochemical technique in which infected cells were detected in formalin-fixed, paraffin-embedded tissue sections using rabbit antiserum to FCV 255, a biotinylated second antibody and streptavidin-peroxidase. Virus was detected by one or both techniques in tonsillar tissues of each animal, and not in other samples. Infected cells were detected in samples from six of eight kittens, and in each animal were few in number, and were cells of the superficial tonsillar epithelium or the stratum germinativum of the adjacent fossa mucosa. Transmission electron microscopic examination of tissues from three of the cats revealed calicivirus-like particles in cells similar to those identified immunohistochemically. These results confirm that the tonsillar region is the major site of FCV persistence and indicate that virus replication during persistence is confined to the surface epithelium of the tonsil and adjacent fossa mucosa.     

Molecular analysis of isolates of feline calicivirus from a population of cats in a rescue shelter.

Two visits, six weeks apart, were made to a cat rescue shelter and single oropharyngeal swabs were taken from all the compliant cats. Feline calicivirus was isolated from 14 of 45 swabs (31 per cent) taken on the first visit and 12 of 46 swabs (26 per cent) taken on the second visit. Nucleotide sequences were obtained for nine isolates from the first visit, six isolates from the second visit, and for the vaccine virus used in the cattery. Distance analysis showed that the majority of the isolates could be assigned to one of two groups. All the isolates obtained from cats sharing the same pen or isolates obtained from the same cat on successive visits, were less than 5 per cent distant, whereas most of the isolates from cats in different pens were more than 20 per cent distant. Phylogenetic analysis showed that at least seven distinct field isolates were present in the cattery. The only good evidence for virus transmission within the cattery was a case in which two viruses isolated from cats in different pens had sequences that were less than 5 per cent distant.