Deletions in the 7a ORF of feline coronavirus associated with an epidemic of feline infectious peritonitis

A population of Persian cats experienced an epidemic of feline infectious peritonitis (FIP) over 2 years. Twelve cases of FIP occurred in litters born during this period. Cats contracting FIP were all genetically related through the sire. Feline coronavirus (FCoV) genomic RNA was detected consistently in this study in biologic samples from adult cats, kittens suffering from FIP, and their siblings. Analysis of viral 7a/7b open reading frame (ORFs) were analyzed and revealed two distinct virus variants circulating in the population, one with an intact 7a ORF and one with two major deletions in the 7a ORF. The 7b ORFs were intact and similar among all virus isolates, although point mutations resulting in amino acid changes were present. The sire was determined to be infected with both variants, and was persistently virus-infected. We speculate the deletion variant arose from the non-deletion variant during viral replication in this population, possibly in the sire.     

An outbreak of feline infectious peritonitis in a Taiwanese shelter: epidemiologic and molecular evidence for horizontal transmission of a novel type II feline coronavirus

Feline infectious peritonitis (FIP) is a fatal disease caused by feline coronavirus (FCoV) infection. FCoV can be divided into serotypes I and II. The virus that causes FIP (FIPV) is believed to occur sporadically and spread infrequently from cat to cat. Recently, an FIP outbreak from an animal shelter was confirmed in Taiwan. FCoV from all the cats in this shelter were analyzed to determine the epidemiology of this outbreak. Thirteen of 46 (28.2%) cats with typical signs of FIP were identified. Among them, seven cats were confirmed by necropsy and/or histopathological examinations. Despite the fact that more than one FCoV was identified in this multi-cat environment, the eight FIP cats were invariably found to be infected with a type II FCoV. Sequence analysis revealed that the type II FIPV detected from fecal samples, body effusions and granulomatous tissue homogenates from the cats that succumbed to FIP all harbored an identical recombination site in their S gene. Two of the cats that succumbed to FIP were found to harbor an identical nonsense mutation in the 3c gene. Fecal shedding of this type II virus in the effusive form of FIP can be detected up to six days before death. Taken together, our data demonstrate that horizontal transmission of FIPV is possible and that FIP cats can pose a potential risk to other cats living in the same environment.     

Evaluation of antibodies against feline coronavirus 7b protein for diagnosis of feline infectious peritonitis in cats

OBJECTIVE: To determine whether expression of feline coronavirus (FCoV) 7b protein, as indicated by the presence of specific serum antibodies, consistently correlated with occurrence of feline infectious peritonitis (FIP) in cats. SAMPLE POPULATION: 95 serum samples submitted for various diagnostic assays and 20 samples from specific-pathogen-free cats tested as negative control samples. PROCEDURES: The 7b gene from a virulent strain of FCoV was cloned into a protein expression vector. The resultant recombinant protein was produced and used in antibody detection assays via western blot analysis of serum samples. Results were compared with those of an immunofluorescence assay (IFA) for FCoV-specific antibody and correlated with health status. RESULTS: Healthy IFA-seronegative cats were seronegative for antibodies against the 7b protein. Some healthy cats with detectable FCoV-specific antibodies as determined via IFA were seronegative for antibodies against the 7b protein. Serum from cats with FIP had antibodies against the 7b protein, including cats with negative results via conventional IFA. However, some healthy cats, as well as cats with conditions other than FIP that were seropositive to FCoV via IFA, were also seropositive for the 7b protein. CONCLUSIONS AND CLINICAL RELEVANCE: Expression of the 7b protein, as indicated by detection of antibodies against the protein, was found in most FCoV-infected cats. Seropositivity for this protein was not specific for the FCoV virulent biotype or a diagnosis of FIP.     

Laboratory changes consistent with feline infectious peritonitis in cats from multicat environments

The present study describes the prevalence of haematological and electrophoretic changes consistent with the diagnosis of feline infectious peritonitis (FIP) in cats without FIP living in six multicat environments with different prevalence of FIP and of other diseases. The results allow designing haematological and electrophoretic profiles typical of each group, most likely depending on the management and on the health status of the group rather than on the prevalence of FIP. In fact, many cats from the colonies with open management and frequent outbreaks of infectious diseases other than FIP had one or more haematological and/or electrophoretical changes consistent with FIP, compared with the reference ranges. In the case of non-specific clinical signs such as fever or neurological signs because of diseases other than FIP, these cats would be erroneously considered as affected by FIP and euthanasized. The use of internal ranges designed on the basis of repeated samplings from non-symptomatic cats allows avoiding these misinterpretations. Results from cats with symptoms consistent with FIP living in the same colonies were also compared with both the reference ranges and the internal ones: such a comparison demonstrated that the use of internal ranges rarely affected the possibility to correctly diagnose the disease in cats with symptoms suggestive of FIP.     

Long-term follow-up study of cats vaccinated with a temperature-sensitive feline infectious peritonitis vaccine.

The long-term safety of a temperature-sensitive feline infectious peritonitis (FIP) vaccine was evaluated. Five hundred eighty-two healthy cats of various age groups were vaccinated with 2 doses of the vaccine. Seventy-eight percent, or 453 cats, were available for follow-up. The mean follow-up period was 541 days. At the end of the follow-up period, 427 cats (94%) were alive. FIP was not diagnosed in any cat during the follow-up period, but 1 cat died of FIP after completion of the follow-up period. Fifty cats (11%) presented with a problem during the follow-up period, but there were typical of those seen in a feline practice. The temperature-sensitive FIP vaccine appears to be safe for use in the general cat population. It does not appear to sensitize cats to develop FIP, nor do there appear to be any other systemic problems associated with use of the vaccine.     

Prevalence of feline coronavirus types I and II in cats with histopathologically verified feline infectious peritonitis

Feline coronaviruses (FCoV) vary widely in virulence causing a spectrum of clinical manifestations reaching from subclinical course to fatal feline infectious peritonitis (FIP). Independent of virulence variations they are separated into two different types, type I, the original FCoV, and type II, which is closely related to canine coronavirus (CCV). The prevalence of FCoV types in Austrian cat populations without FIP has been surveyed recently indicating that type I infections predominate. The distribution of FCoV types in cats, which had succumbed to FIP, however, was fairly unknown. PCR assays have been developed amplifying parts of the spike protein gene. Type-specific primer pairs were designed, generating PCR products of different sizes. A total of 94 organ pools of cats with histopathologically verified FIP was tested. A clear differentiation was achieved in 74 cats, 86% of them were type I positive, 7% type II positive, and 7% were positive for both types. These findings demonstrate that in FIP cases FCoV type I predominates, too, nonetheless, in 14% of the cases FCoV type II was detected, suggesting its causative involvement in cases of FIP.     

Ischaemic encephalopathy and focal granulomatous meningoencephalitis in the cat

Three cats were presented with neurological deficits compatible with cerebral disease. Two of the cats had temporal lobe infarction diagnosed as ischaemic encephalopathy; one cat had granulomatous inflammation, diagnosed as focal granulomatous meningoencephalitis. In all three cases the lesions were characterized by necrotic, cavitated areas. Vascular insult was the cause of the ischaemic encephalopathy and the focal granulomatous meningoencephalitis may have been caused by feline infectious peritonitis (FIP) virus. An infectious aetiology, possibly FIP virus, is proposed for the ischaemic encephalopathy. A relationship between these two neurological diseases may exist.     

Risk factors for feline infectious peritonitis in Australian cats

The objective of this study was to determine whether patient signalment (age, breed, sex and neuter status) is associated with naturally-occurring feline infectious peritonitis (FIP) in cats in Australia. A retrospective comparison of the signalment between cats with confirmed FIP and the general cat population was designed. The patient signalment of 382 FIP confirmed cases were compared with the Companion Animal Register of NSW and the general cat population of Sydney. Younger cats were significantly over-represented among FIP cases. Domestic crossbred, Persian and Himalayan cats were significantly under-represented in the FIP cohort, while several breeds were over-represented, including British Shorthair, Devon Rex and Abyssinian. A significantly higher proportion of male cats had FIP compared with female cats. This study provides further evidence that FIP is a disease primarily of young cats and that significant breed and sex predilections exist in Australia. This opens further avenues to investigate the role of genetic factors in FIP.     

Genetic determinants of pathogenesis by feline infectious peritonitis virus

Feline infectious peritonitis (FIP) is a fatal, immune-augmented, and progressive viral disease of cats associated with feline coronavirus (FCoV). Viral genetic determinants specifically associated with FIPV pathogenesis have not yet been discovered. Viral gene signatures in the spike, non-structural protein 3c, and membrane of the coronavirus genome have been shown to often correlate with disease manifestation. An "in vivo mutation transition hypothesis" is widely accepted and postulates that de novo virus mutation occurs in vivo giving rise to virulence. The existence of "distinct circulating avirulent and virulent strains" is an alternative hypothesis of viral pathogenesis. It may be possible that viral dynamics from both hypotheses are at play in the occurrence of FIP. Epidemiologic data suggests that the genetic background of the cat contributes to the manifestation of FIP. Further studies exploring both viral and host genetic determinants of disease in FIP offer specific opportunities for the management of this disease.     

Retrospective analysis of seizures associated with feline infectious peritonitis in cats

Seizures have been reported frequently in feline infectious peritonitis (FIP) but have not been studied in detail in association with this disease. The purpose of this study was to perform a retrospective analysis of neurological signs in a population of 55 cats with a histopathologically confirmed neurological form of FIP. Seizure patterns were determined and it was attempted to relate occurrence of seizures with age, breed, sex and neuropathological features. Fourteen cats had seizure(s), while 41 cats had no history of seizure(s). Generalised tonic-clonic seizures were seen in nine cats; and complex focal seizures were observed in four patients. The exact type of seizure could not be determined in one cat. Status epilepticus was observed in one patient but seizure clusters were not encountered. Occurrence of seizures was not related to age, sex, breed or intensity of the inflammation in the central nervous system. However, seizures were significantly more frequent in animals with marked extension of the inflammatory lesions to the forebrain (P=0.038). Thus, the occurrence of seizures in FIP indicates extensive brain damage and can, therefore, be considered to be an unfavourable prognostic sign.     

Identification and genotyping of feline infectious peritonitis-associated single nucleotide polymorphisms in the feline interferon-γ gene

Feline infectious peritonitis (FIP) is an immune-mediated, highly lethal disease caused by feline coronavirus (FCoV) infection. Currently, no protective vaccine or effective treatment for the disease is available. Studies have found that some cats survive the challenge of virulent FCoV isolates. Since cellular immunity is thought to be critical in preventing FIP and because diseased cats often show a significant decrease in interferon-γ (IFN-γ) production, we investigated whether single nucleotide polymorphisms (SNP) in the feline IFN-γ gene (fIFNG) are associated with the outcome of infection. A total of 82 asymptomatic and 63 FIP cats were analyzed, and 16 SNP were identified in intron 1 of fIFNG. Among these SNP, the fFING + 428 T allele was shown to be a FIP-resistant allele (p = 0.03), and the heterozygous genotypes 01C/T and +408C/T were found to be FIP-susceptible factors (p = 0.004). Furthermore, an fIFNG + 428 resistant allele also showed a clear correlation with the plasma level of IFN-γ in FIP cats. For the identification of these three FIP-related SNP, genotyping methods were established using amplification refractory mutation system PCR (ARMS-PCR) and restriction fragment length polymorphisms (RFLP), and the different genotypes could easily be identified without sequencing. The identification of additional FIP-related SNP will allow the selection of resistant cats and decrease the morbidity of the cat population to FIP.     

Polymorphisms in the feline TNFA and CD209 genes are associated with the outcome of feline coronavirus infection

Feline infectious peritonitis (FIP), caused by feline coronavirus (FCoV) infection, is a highly lethal disease without effective therapy and prevention. With an immune-mediated disease entity, host genetic variant was suggested to influence the occurrence of FIP. This study aimed at evaluating cytokine-associated single nucleotide polymorphisms (SNPs), i.e., tumor necrosis factor alpha (TNF-α), receptor-associated SNPs, i.e., C-type lectin DC-SIGN (CD209), and the five FIP-associated SNPs identified from Birman cats of USA and Denmark origins and their associations with the outcome of FCoV infection in 71 FIP cats and 93 FCoV infected non-FIP cats in a genetically more diverse cat populations. A promoter variant, fTNFA - 421 T, was found to be a disease-resistance allele. One SNP was identified in the extracellular domain (ECD) of fCD209 at position +1900, a G to A substitution, and the A allele was associated with FIP susceptibility. Three SNPs located in the introns of fCD209, at positions +2276, +2392, and +2713, were identified to be associated with the outcome of FCoV infection, with statistical relevance. In contrast, among the five Birman FIP cat-associated SNPs, no genotype or allele showed significant differences between our FIP and non-FIP groups. As disease resistance is multifactorial and several other host genes could involve in the development of FIP, the five genetic traits identified in this study should facilitate in the future breeding of the disease-resistant animal to reduce the occurrence of cats succumbing to FIP.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0123-6) contains supplementary material, which is available to authorized users.     

Disease outcome and cytokine responses in cats immunized with an avirulent feline infectious peritonitis virus (FIPV)-UCD1 and challenge-exposed with virulent FIPV-UCD8

Eight cats were immunized with an avirulent strain of feline infectious peritonitis virus (FIPV)-UCD1, then challenge-exposed to a highly virulent cat passaged strain (FIPV-UCD8). Th1 and Th2 cytokine profiles in the peripheral blood mononuclear cells (PBMCs) were measured throughout in the experiment. No clinical signs of FIP were evident in the experimental cats after immunization. After challenge, the immunized cats demonstrated one of four clinical outcomes: (1) classical effusive FIP; (2) accelerated FIP; (3) non-effusive FIP, or (4) resistance to challenge. Only minor cytokine changes were observed following immunization, however, several cytokine changes occurred following challenge-exposure. The most noteworthy changes were in tumor necrosis factor-alpha (TNF-alpha) and interferon gamma (IFN-gamma) levels. Our preliminary findings suggest that immunity against FIP is associated with TNF-alpha and IFN-gamma response imbalance, with high TNF-alpha/low IFN-gamma mRNA responses favouring disease and low TNF-alpha/high IFN-gamma mRNA responses being indicative of immunity.     

Mutations in feline immunodeficiency (FIV) virus envelope gene V3-V5 regions in FIV-infected cats

The envelope (Env) gene V3-V5 regions of the feline immunodeficiency virus (FIV) encode the neutralizing epitopes. Since mutations in these regions induce resistance to viral neutralizing antibodies, they may influence the effects of vaccines. To examine the in vivo mutation rate in these regions, we cloned cDNA for the Env gene V3-V5 regions from the PBMC of experimentally FIV-infected cats, and compared the deduced amino acid sequences. Blood or plasma from an FIV Shizuoka strain-infected cat was inoculated into a second group of SPF cats, and their blood or plasma was inoculated into the third group. The amino acid sequence encoded by the viral gene of the first cat was compared with those encoded by the viral genes of a total of eight cats in the second and third groups (two and six cats, respectively). The amino acid sequences in two cats in the second and third groups were 100% homologous and in one cat in the third group was 98.3% homologous to that in the first infected cat. Five cats had the same sequence, which was 97.8% homologous to that in the first infected cat. Three kittens, born 2 months after the inoculation of the FIV Aomori-2 strain into the mother cat, were anti-FIV negative at 4 weeks after birth, but became seropositive at 33 weeks after birth, confirming FIV infection. Comparison of the encoded amino acid sequences of the viral gene in two cats at 48 weeks after birth showed 100% homology to that of the virus inoculated into the mother cat, and the remaining one cat had a single residue substitution, resulting in 99.4% homology. These results suggest that the FIV Env gene V3-V5 regions are stably maintained for at least 1-2 years after infection.     

Effect of feline interferon-omega on the survival time and quality of life of cats with feline infectious peritonitis

BACKGROUND: There is no therapy with proven efficacy to treat cats with feline infectious peritonitis (FIP). HYPOTHESIS: Feline interferon-omega (FeIFN-omega) prolongs survival time and increases quality of life in cats with FIP. ANIMALS: Thirty-seven privately owned cats were subjects of this study. METHODS: The study was performed as a placebo-controlled double-blind trial. Feline infectious peritonitis was confirmed by histology or immunostaining of feline coronavirus (FCoV) antigen in effusion or tissue macrophages or both. The cats were randomly selected for treatment with either FeIFN-omega or a placebo. All cats received adjunctive treatment with glucocorticoids and antibiotics and passive immunization with Feliserin. RESULTS: There was no statistically significant difference in the survival time of cats treated with FeIFN-omega versus placebo or in any other variable evaluated (with the exception of the lymphocyte count). The cats survived between 3 and 200 days (median, 9 days). There was only 1 long-term survivor (> 3 months), and the cat was in the FeIFN-omega group. CONCLUSION AND CLINICAL RELEVANCE: No effect of FeIFN-omega on survival time or quality of life could be demonstrated in this study.     

Prevalence of feline infectious peritonitis in specific cat breeds

Although known that purebreed cats are more likely to develop feline infectious peritonitis (FIP), previous studies have not examined the prevalence of disease in individual breeds. All cats diagnosed with FIP at a veterinary teaching hospital over a 16-year period were identified. Breed, sex and reproductive status of affected cats were compared to the general cat population and to mixed breed cats evaluated during the same period. As with previous studies sexually intact cats and purebreed cats were significantly more likely to be diagnosed with FIP; males and young cats also had a higher prevalence of disease. Abyssinians, Bengals, Birmans, Himalayans, Ragdolls and Rexes had a significantly higher risk, whereas Burmese, Exotic Shorthairs, Manxes, Persians, Russian Blues and Siamese cats were not at increased risk for development of FIP. Although additional factors doubtlessly influence the relative prevalence of FIP, this study provides additional guidance when prioritizing differentials in ill purebreed cats.     

Clinicopathological findings and disease staging of feline infectious peritonitis: 51 cases from 2003 to 2009 in Taiwan

Fifty-one cats histopathologically confirmed to have been naturally infected by feline infectious peritonitis (FIP), were collected to analyse the clinical and laboratory findings and to characterise disease staging. Effusive FIP was found in 33 cats, non-effusive FIP in 12 cats, and mixed-type in six cats. Highly significant decreases in haematocrit and albumin levels and an increase in total bilirubin level were noted in both effusive and non-effusive FIP, at first presentation and before death. In serial blood examinations of the effusive group, anaemia and increases in bilirubin and aspartate aminotransferase (AST) were observed from 2 weeks to 0-3 days before death. The packed cell volume, bilirubin, AST, potassium, and sodium levels were established to predict disease staging and survival time. Cumulative points ranging from 0 to 4, 5 to 11 and excess of 12, indicate that the cat can survive for at least 2 weeks, less than 2?weeks and less than 3 days, respectively.