Chronic oral infections of cats and their relationship to persistent oral carriage of feline calici-, immunodeficiency, or leukemia viruses.

Two hundred and twenty-six cats from the Veterinary Medical Teaching Hospital (VMTH), a cat shelter, and a purebred cattery were tested for chronic feline calicivirus (FCV), feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) infections. Chronic oral carriage of FCV was present in about one-fifth of the cats in each of the groups. FIV infection was not present in the purebred cattery, was moderately prevalent (8%) in the pet population of cats examined at the VMTH for various complaints and was rampant in the cat shelter (21%). Unexpectedly high FeLV infection rates were found in the hospital cat population (28%) and in the purebred cattery (36%), but not in the cat shelter (1.4%). FCV and FeLV infections tended to occur early in life, whereas FIV infections tended to occur in older animals. From 43 to 100% of the cats in these environments had oral cavity disease ranging from mild gingivitis (23-46%), proliferative gingivitis (18-20%), periodontitis (3-32%) and periodontitis with involvement of extra-gingival tissues (7-27%). Cats infected solely with FCV did not have a greater likelihood of oral lesions, or more severe oral disease, than cats that were totally virus free. This was also true for cats infected solely with FeLV, or for cats dually infected with FeLV and FCV. Cats infected solely with FIV appeared to have a greater prevalence of oral cavity infections and their oral cavity disease tended to be more severe than cats without FIV infection. FIV-infected cats that were coinfected with either FCV, or with FCV and FeLV, had the highest prevalence of oral cavity infections and the most severe oral lesions.     

Shedding of feline immunodeficiency virus in semen of domestic cats during acute infection

OBJECTIVE: To examine shedding of cell-free and cell-associated feline immunodeficiency virus (FIV) in semen of domestic cats during acute infection. ANIMALS: 7 specific-pathogen-free sexually intact male cats. PROCEDURE: 6 cats were inoculated IV with 5 x 10(6) 50% tissue culture infective doses of FIV-NCSU1, and 1 cat served as an uninfected (control) cat. Infection was confirmed in the 6 cats. Periodically for up to 16 weeks after inoculation, cats were anesthetized and ejaculates obtained by use of electroejaculation. Virus was isolated from filtered seminal plasma and washed seminal cells by co-cultivation with a feline CD4+ T-cell line. Seminal cell lysates were also examined for a 582-base pair segment of FIV gag provirus DNA, using a nested polymerase chain reaction amplification. RESULTS: During the acute phase of FIV infection, virus was evident in semen of 5 inoculated cats. Five cats had virus-positive seminal plasma and 3 had virus-positive cellular constituents during the study. Virus was isolated from 8/22 (36%) seminal plasma samples and 2/17 (18%) seminal cell specimens. Provirus DNA was detected in 5/24 (21%) seminal cell lysates. Cell-free virus was isolated as early as 6 weeks after inoculation, whereas cell-associated virus was isolated as early as 12 weeks after inoculation. Provirus DNA was detected in seminal cells from one cat as early as 1 week after inoculation. CONCLUSIONS AND CLINICAL RELEVANCE: Cell-free and cell-associated FIV are shed in semen of cats early during the course of infection. Samples obtained before seroconversion may contain virus. Virus shedding in ejaculates varies between and within cats during acute infection.     

In vitro activity of VEGF-E produced by orf virus strains isolated from classical and severe persistent contagious ecthyma

Proliferative orf virus infections in adult sheep have increased in Italy in the past few years: these extreme cases are frequently fatal and difficult to differentiate from other infectious diseases of sheep such as blue tongue. A probable explanation for the proliferative and highly vascularized nature of the lesions was found in the expression of the VEGF-E gene encoded by the orf virus. To investigate a possible role of the viral VEGF in the pathogenesis of severe persistent orf virus lesions, the activity of four VEGF-E variants was compared by an angiogenesis in vitro model. Similar angiogenic activity was found between strains isolated from the classical and the proliferative forms of the disease, even if the latter was able to develop a higher number of vessels during the first 24 h of infection. Our in vitro findings seems to exclude that the VEGF variants encoded by the strain isolated from the atypical form of the disease could be the responsible for the histopathological aspect of the proliferative lesions.     

In vivo T-cell subset depletion suggests that CD4+ T-cells and a humoral immune response are important for the elimination of orf virus from the skin of sheep

In vivo lymphocyte subset depletion offers a unique opportunity to study the roles of different cellular components of the immune system of sheep during infection with orf virus. Lambs were depleted of specific lymphocyte subsets by the intravenous administration of monoclonal antibodies against ovine lymphocyte surface markers and then challenged with orf virus. The skin lesions that developed were scored visually as to their severity. Blood samples were collected to monitor the lymphocyte depletions and to measure orf-virus-specific antibody levels. Skin biopsies were collected from the lesion site and studied to determine the course of the infection and the presence of various cell types and orf virus.All the sheep developed orf virus lesions after infection. All three of the CD4-depleted lambs were unable to clear virus from their skin and did not have an antibody response to the virus. Virus was also detected in the skin of one each of the three CD8-depleted, WC1-depleted and control sheep on the final day of the trial. CD8(+) lymphocytes did not appear to be essential for viral clearance later in the infection. Depletion of the majority of gammadelta(+) T-cells did not affect the outcome of orf virus infection. In sheep with high orf-virus-specific antibody titres at the time of infection, orf lesions healed faster than lesions in sheep with low antibody levels, and this occurred regardless of the lymphocyte depletion status of the animals.This study suggests that the presence of CD4(+) T-cells and orf-virus-specific antibodies are important for the control of viral replication in the skin of infected sheep.     

PATHOGENICITY OF A STRAIN OF FELINE CALICIVIRUS FOR DOMESTIC KITTENS

A strain of feline calicivirus, isolated from a cheetah exhibiting ulcerative glossitis and conjunctivitis, was administered by aerosol to 4 domestic cats and by contact to a fifth cat. Despite the limited number of animals available for the experiment, the pathogenicity of the virus strain for domestic cats was established. In aerosol-infected animals, clinical signs were referable to infection of both upper and lower respiratory tracts. The virus produced an interstitial pneumonia which, early in infection, was uncomplicated by secondary bronchopneumonia. The in-contact cat exhibited clinical signs referable to infection of the oral cavity only and no lesions were noted in the lower respiratory tract at autopsy. Ulcerative glossitis was a feature of the disease in aerosol-infected and in-contact cats. The virus was isolated from the pharynx of all cats for up to 21 days after infection and from the tonsils at autopsy. The tonsils were considered to be a probable site of multiplication of virus in "carrier" cats.     

Severe persistent orf in young goats.

Orf (contagious ecthyma) is a viral disease of small and wild ruminants, humans, and less frequently other species. In sheep and goats, the disease is characterized by the formation of vesiculo-proliferative lesions in the skin of lips and nostril. Here, a form of generalized orf in 16 goat kids from 2 different locations in west Texas is described. The disease was characterized by multifocal, severe, proliferative dermatitis that persisted from about 2 months of age until the goat kids were euthanized 3 months later. All affected goats were Boer or Boer crosses under 1 year of age. The mean immunoglobulin concentration in sera of affected goats was elevated compared with healthy control goats. Severe to moderate lymphadenomegaly of the nodes draining the areas of the skin affected with orf lesions was present in all 16 goat kids. Suppurative arthritis, chronic fibrinous pneumonia, and premature thymic involution were found in 3, 5, and 7 of the goat kids, respectively. The skin lesions of 3 goat kids were infested with larvae of the opportunistic black garbage fly (Ophira sp.). The orf virus was identified in skin lesions by isolation in Marbin-Darby ovine kidney cells, electron microscopy, and amplification of viral DNA by polymerase chain reaction. The orf virus was not detected in peripheral blood or lymph node mononuclear cells of any of the goats. Cross-neutralization experiments showed that an ovine orf virus antiserum raised in sheep was more effective in neutralizing a sheep orf virus isolate than a caprine orf virus isolate. The clinical and epidemiological characteristics of these orf cases may be the result of susceptibility factors within some individuals of the Boer breed of goats.     

Natural and experimental pox virus infection in the domestic cat

Three naturally occurring cases of cowpox virus infection in the domestic cat are described. Isolate L97 was identified as cowpox virus on the basis of morphology, serology and characteristic cytopathic effect in tissue culture and on the chorioallantoic membrane of embryonated eggs. All three cases showed multiple skin lesions, slight conjunctivitis or purulent ocular discharge but there were no respiratory signs. Two animals recovered, the third was put down as a stray. The disease was reproduced in experimental cats. Isolate L97 was inoculated into two cats intravenously and two cats by skin scarification. All four developed skin lesions at the site or sites of inoculation, and in one cat multiple lesions developed. The two intravenously inoculated animals also developed severe oedema of the neck and brisket around the site of inoculation into the jugular vein, and one cat died. Serological and pathological findings on both the natural and experimental infections are described. Serum neutralising antibody titres in both natural and experimental early convalescent cases were significantly enhanced by the addition of complement.     

The development of oral lesions in lambs naturally infected with orf virus

Stomatitis in sheep caused by orf virus can be confused with lesions of more economically significant diseases, including foot-and-mouth disease, but there is no published account of the sequential development of oral orf lesions in the sheep. This report describes the clinical appearance of such lesions during a natural outbreak of the disease in young lambs. Lesions were seen on the gingiva, the tongue and the dental pad/hard palate, and progressed from small erythematous papules to larger, often coalescing papules that in some cases were ulcerated. Resolution started within seven days and was complete within 22 days. The lambs continued to suck and thrive throughout the infection. Lesions at all stages were proliferative, providing a major differentiating factor between orf and other causes of stomatitis in sheep.     

Evaluation of praziquantel for treatment of experimentally induced paragonimiasis in dogs and cats

Praziquantel was used successfully for treatment of a small number of dogs and 1 cat infected with Paragonimus kellicotti. To further evaluate the usefulness of this drug in treating such infections, 7 cats and 7 dogs were inoculated orally with metacercariae (12 and 20 to 22, respectively) obtained from crayfish, then were treated after the infections became patent; 2 cats and 2 dogs served as noninfected controls. Beginning 1 week before infection, and continuing weekly thereafter, physical, hematologic, and fecal examinations were performed on each animal; thoracic radiography was performed every other week. By postinoculation week 6, all dogs given metacercariae had patent infection diagnosed on the basis of positive results of fecal examination. By postinoculation week 7, 5 cats had confirmed patent infection, but 2 cats given metacercariae never had patent infection or had signs of infection. Clinical signs of infection were minor and included increased respiratory tract noise, slight inducible cough, or mild dyspnea. Transient eosinophilia was detected in dogs around postinoculation week 3. Pretreatment radiography revealed cavitated lesions in cats only; pleural lines and patchy infiltrates in cats and dogs; or pneumothorax in dogs only. The treatment regimen consisted of 23 mg of praziquantel/kg of body weight given every 8 hours for 3 days; 1 infected cat and dog were not treated. By 11 days after treatment, eggs had disappeared from the feces of infected animals, and marked resolution of lung lesions was evident radiographically.(ABSTRACT TRUNCATED AT 250 WORDS)     

A severe outbreak of contagious ecthyma (orf) in a free-ranging musk ox (Ovibos moschatus) population in Norway

During July-October 2004, 19 (18 calves, 1 yearling) free-ranging musk oxen (Ovibos moschatus) at Dovre, Norway, were observed with contagious echtyma-like lesions, and 16 of them were euthanized. Six musk oxen were subjected to necropsy, histopathological and microbiological examinations. All euthanized animals had lesions consistent with contagious ecthyma presenting as wart-like, scabby lesions on the muzzle, lips, oral mucosa and limbs to a variable extent. The histopathological examination showed pustular dermatitis characterized by epidermal proliferation, reticular degeneration, degenerating keratinocytes with intracytoplasmic eosinophilic inclusion bodies, vesicopustules, microabscesses and multifocal ulcerations in the epidermis which was covered by a serocellular crust. Pathology and bacteriology showed evidence of secondary infections in the skin and draining lymph nodes. Electron microscopy (negative staining) of lesions from four animals detected parapoxvirus with the typical arrangement of the outer protein filaments. Parapoxvirus DNA was detected in tissue samples from two examined animals by polymerase chain reaction (PCR) with primers from the B2L-gene. A DNA sequence of 326 nucleotides from the amplicon was compared with similar DNA sequences from parapoxvirus isolated from sheep, reindeer, musk ox and cattle. The outbreak was caused by a virus similar to other circulating orf virus variants in Norway. Antibodies against parapoxvirus were detected with a virus neutralization test in 3 of 35 musk oxen (8.6%) sampled at Dovre between 2004 and 2006. This is the first report of a severe outbreak of contagious ecthyma in free-ranging musk oxen.     

Susceptibility of cats, sheep, and swine to a rabbit isolate of Encephalitozoon cuniculi

Newborn cats, pigs, and sheep (3 to 14 days old) and postweanling cats (2.5 months old) that had been inoculated with Gardner feline sarcoma virus and feline leukemia virus at 10 days of age were infected experimentally with a rabbit isolate of the mammalian protozoan parasite Encephalitozoon cuniculi. Infection occurred in all cats and in some sheep, but was questionable in pigs. Brain and kidney were the 2 major target organs in cats. The lesions were compatible with, but less severe than, those of naturally infected cats and other carnivores. Of 13 cats, E cuniculi could be detected morphologically in the kidneys of 12 cats and in the brain of 1 cat. The organisms were reisolated from 2 cats with ground tissue suspension of kidney or urine sediment. The indirect immunofluorescence antibody (IFA) titers were 1:20 to 1:1,280 at the time the animals were killed, but antibodies were not detected before inoculation. Lesions were seen in the kidneys of 2 of 4 sheep. These lesions were mild, but were compatible with those in a spontaneously affected goat. Encephalitozoon cuniculi were found morphologically in the kidney of 1 sheep with lesions. All sheep had IFA titers of 1:10 to 1:20 before inoculation, and the titers were 1:20 to 1:320 when they were killed. Vasculitis, similar to the subacute-to-chronic stage of polyarteritis nodosa, was observed in 1 of 8 pigs. The lesions were primarily present in the kidney; comparable but milder lesions were also seen in the heart and brain. Antibody was not detected before inoculation.(ABSTRACT TRUNCATED AT 250 WORDS)     

Genetic characterization of orf viruses isolated from various ruminant species of a zoo

In the present study, an outbreak of proliferative dermatitis in musk ox (Ovibos moschatus), Sichuan takin (Budorcas taxicolor tibetana) and domestic Shetland sheep (Ovis aries) in a zoo is described. Skin lesions consisted of severe, persistent, multifocal, proliferative dermatitis in musk ox, and mild, transient, focal, dermatitis in the Sichuan takin and Shetland sheep. Parapoxviruses were isolated from skin lesions, and characterized by restriction enzyme analysis and partial gene sequencing. The results of this investigation indicate that the outbreak of proliferative dermatitis was due to infection by a single parapoxvirus, which is genetically closely related to other orf virus (ORFV) strains but distant to bovine papular stomatitis virus (BPSV) and pseudocowpox virus (PCPV).     

Ulcerative facial and nasal dermatitis and stomatitis in cats associated with feline herpesvirus 1

Ulcerative dermatitis of the nasal planum or haired skin of the face, associated with intranuclear inclusion bodies compatible with herpesvirus, was identified in nine cats. Clinically, lesions were ulcerative and crusted, and often persistent. A tenth cat had focal proliferative ulcerative stomatitis, also associated with intranuclear inclusion bodies. Microscopically, there was necrosis and ulceration associated with prominent eosinophilic inflammation. Intranuclear inclusion bodies were noted in all cases, within the surface or adnexal epithelium. Ultrastructural examination of skin from two cats revealed virions morphologically compatible with a herpesvirus. Polymerase chain reaction (PCR) specific for feline herpesvirus 1 on DNA extracted from fresh-frozen or formalin-fixed paraffin-embedded biopsy samples and/or consensus primer PCR with DNA sequencing performed on DNA extracted from formalin-fixed paraffin-embedded biopsy samples from seven cats revealed that the virus was indistinguishable from feline herpesvirus 1. PCR was negative in one of eight cats tested.     

Is feline foamy virus really apathogenic?

Feline foamy virus (FFV) is a retrovirus commonly found in cats. It is generally thought to be apathogenic, making it a suitable candidate as a gene therapy vector. However, there have been reports of association of FFV with chronic progressive arthritis and a cofactor effect with feline immunodeficiency virus. This study investigated experimental FFV infection and whether this was associated with signs of disease.

Eight young specific pathogen free cats were inoculated intramuscularly with FFV. The cats were examined twice weekly and blood and pharyngeal samples were taken. Haematology, biochemistry and FFV quantitative polymerase chain reaction (qPCR) were performed. Tissue samples were also collected throughout the six month period.

FFV was initially detected by qPCR in the blood within the first two weeks of infection and viraemia persisted throughout the study. Two peaks of viraemia were observed, at day 20 (80–170 FFU/ml blood) and day 155 (332–415 FFU/ml blood). FFV was also consistently detected in oropharyngeal samples after day 36. Anti-FFV IgG was detected in all cats by ELISA; antibody levels had an early peak around day 35 and then increased again following the second rise in circulating viral load. All cats remained clinically normal, except for one cat with an unrelated gingivitis. None of the cats developed pyrexia. The biochemical profile and blood cell counts remained within normal limits except for one cat with a persistent eosinophilia. Initial fluctuations in white cell counts settled within three weeks and did not deviate outside of the normal ranges. All tissue samples contained FFV DNA; lymphoreticular tissues, salivary gland and lung had the highest viral loads. Although there were no gross pathological lesions on post mortem examination, histologically a mild glomerulonephritis and a moderate interstitial pneumonia were observed in all cats.

We conclude that during the six month period of infection, although cats appeared clinically normal, histopathological changes were observed in the lungs and kidneys. Further investigation of the significance of these changes is warranted before FFV is developed as a vector for gene delivery.     

Detection of cellular cytokine mRNA expression during orf virus infection in sheep: differential interferon-gamma mRNA expression by cells in primary versus reinfection skin lesions.

In sheep infected with the parapoxvirus orf virus, primary infection orf skin lesions developed and resolved within 8 weeks. Reinfection lesions were smaller and resolved within 3 weeks. The host response in the skin was characterized by an accumulation of neutrophils, dendritic cells, CD4+ T cells, CD8+ T cells, B cells and T19+ gammadelta T cells. The magnitude of this accumulation paralleled orf virus replication in the skin. In situ hybridization was used to detect cells expressing interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha) and interleukin-4 (IL-4) mRNAs in orf skin. Cells expressing IL-4 mRNA were not detected at any time after infection. Cells expressing IFN-gamma mRNA were detected after reinfection but not after primary infection. Cells expressing TNF-alpha mRNA included epidermal cells, vascular endothelium and uncharacterized cells that increased more rapidly in the skin after reinfection compared to primary infection. The results are consistent with a prominent role for IFN-gamma in the host immune response controlling the severity of the disease.     

Long-term impact on a closed household of pet cats of natural infection with feline coronavirus, feline leukaemia virus and feline immunodeficiency virus

A closed household of 26 cats in which feline coronavirus (FCoV), feline leukaemia virus (FeLV) and feline immunodeficiency virus (FIV) were endemic was observed for 10 years. Each cat was seropositive for FCoV on at least one occasion and the infection was maintained by reinfection. After 10 years, three of six surviving cats were still seropositive. Only one cat, which was also infected with FIV, developed feline infectious peritonitis (FIP). Rising anti-FCoV antibody titres did not indicate that the cat would develop FIP. The FeLV infection was self-limiting because all seven of the initially viraemic cats died within five years and the remainder were immune. However, FeLV had the greatest impact on mortality. Nine cats were initially FIV-positive and six more cats became infected during the course of the study, without evidence of having been bitten. The FIV infection did not adversely affect the cats' life expectancy.     

Hematologic and radiographic changes in cats after inoculation with infective larvae of Dirofilaria immitis.

Hematologic and radiographic findings in 15 domestic cats inoculated with 25, 100, or 200 to 400 infective larvae of Dirofilaria immitis and in 5 uninoculated cats were compared. Eosinophilia and leukocytosis developed 3 to 5 months after inoculation; packed cell volume and hemoglobin values were similar for inoculated cats and controls. Changes detected in radiographs of inoculated cats were: (1) increased visualization of the pulmonary arteries; (2) right-side cardiac enlargement; and (3) diffuse to focal areas of density in the pulmonary parenchyma. At least one of these lesions was visible radiographically by 3 to 7 months after inoculation in all 15 cats. The severity of changes was not directly proportional to the number of infective larvae inoculated, adult worm burden, or age of cat. However, male cats had more severe cardiopulmonary lesions than did females, and cats that had microfilaremia during the course of infection had more severe pulmonary lesions, as determined radiographically. Parenchymal densities decreased in 8 of the 15 inoculated cats 6 to 14 months after inoculation. Cardiac enlargement was detected radiographically in 10 of the inoculated cats. Enlargement of the pulmonary arteries was the most consistent sign of D immitis infection; it developed and persisted in 12 cats found to be infected at necropsy.     

Isolation of parapoxvirus from a cow treated with interferon-gamma

A virus was isolated from peripheral blood leukocytes of a cow which was kept in an isolated pen after it was injected with recombinant bovine interferon-γ. The virus was identified as a member of genus Parapoxvirus in the family Poxviridae on the basis of electron microscopic observations and serological tests. Parapoxvirus has seldom been isolated other than from papular lesions, the characteristic sign of parapoxvirus infection. This is the first report of parapoxvirus isolation from the peripheral blood of a cow without any clinical signs. These results show that parapoxviruses are capable of causing persistent infection in cattle without clinical signs and can be activated by stress factors that induce modification of immune reactions. Relationships between the isolated virus and other parapoxviruses isolated previously from cattle in Japan were investigated and discussed.     

Response of efferent lymph and popliteal lymph node to epidermal infection of sheep with orf virus

Functional and phenotypic changes in the cell populations were monitored in the popliteal efferent lymph of sheep following experimental epidermal infection with orf virus. In another group of sheep, cells from the popliteal lymph node draining the site of infection were similarly monitored and compared with the cells from contralateral popliteal and mesenteric lymph nodes. All sheep showed serological evidence of previous exposure to orf virus. Following infection, anti-orf antibody titres rose and efferent lymphocyte and blast cell output increased. Interferon-like activity was detected in efferent lymph early after orf virus but not mock infection. Lymphocytes from the draining popliteal lymph node showed antigen-specific lymphoproliferation on Days 3-7 while cells in the efferent lymph demonstrated proliferative activity on Days 4-6. The requirement for exogenous antigen-presenting cells in the culture of efferent lymphocytes varied between individual sheep. The culture supernatant from proliferating lymph node cells contained interferon-like activity but no anti-orf antibodies, the reverse of that from cultured efferent lymphocytes, perhaps indicating a different reactive T cell population. During the course of the experiment there was an increase in the percentage of efferent lymphocytes expressing MHC Class II antigens and surface immunoglobulins, the latter being recorded as a double peak. The short-term nature of the local T cell response may in part explain the incompleteness of immunity to orf virus in sheep.