Sylvia J. Tucker

Enteropathogens identified in cats entering a Florida animal shelter with normal feces or diarrhea

OBJECTIVE To determine the frequency of enteropathogens in cats entering an animal shelter with normal feces or diarrhea. DESIGN Cross-sectional study. ANIMALS 100 cats evaluated at an open-admission municipal animal shelter in Florida. PROCEDURES Fecal samples collected within 24 hours after admission from 50 cats with normal feces and 50 cats with diarrhea were tested by fecal flotation, antigen testing, PCR assay, and electron microscopy for selected enteropathogens. RESULTS 12 enteropathogens were identified. Cats with diarrhea were no more likely to be infected with ≥ 1 (84%) enteropathogens than were cats with normal feces (84%). Only feline coronavirus was significantly more prevalent in cats with diarrhea (58%) than in cats with normal feces (36%). Other enteropathogens identified in cats with and without diarrhea included Clostridium perfringens enterotoxin A (42% and 50%, respectively), Cryptosporidium spp (10% and 20%, respectively), Giardia spp (20% and 8%, respectively), Cystoisospora spp (14% and 10%, respectively), hookworms (10% and 18%, respectively), ascarids (6% and 16%, respectively), Salmonella spp (6% and 4%, respectively), astrovirus (8% and 2%, respectively), feline panleukopenia virus (4% and 4%, respectively), calicivirus (0% and 2%, respectively), and Spirometra spp (0% and 2%, respectively). CONCLUSIONS AND CLINICAL RELEVANCE In the present study, cats entered the shelter with a variety of enteropathogens, many of which are pathogenic or zoonotic. Most infections were not associated with diarrhea or any specific risk factors such as signalment, source, or body condition, making it difficult to predict which cats were most likely to be infected. It is not possible to test all shelter cats for all possible infections, so practical guidelines should be developed to treat routinely for the most common and important enteropathogens.

Infectious diseases of dogs and cats on Isabela Island, Galapagos.

Background: Vaccination and importation of dogs and cats are prohibited in the Galapagos, resulting in a uniquely isolated population. The purpose of this study was to determine the prevalence of infectious diseases of dogs and cats that impact their health, could spill over to native wildlife, or sentinel diseases of concern to humans. Hypothesis: The isolation of dogs and cats in the Galapagos protects them from diseases common in mainland populations. Animals: Ninety‐five dogs and 52 cats presented during a neutering campaign. Methods: A prospective cross‐sectional study was performed. Blood was collected for serological and DNA evaluation of a panel of infectious diseases. Results: Antibodies against parvovirus (100%), parainfluenza virus (100%), adenovirus 1/2 (66–67%), and distemper virus (22%) were present in dogs. Dirofilaria immitis was also common in dogs (34%), with lower prevalences of Wolbachia pipiens (22%), Bartonella sp. (13%), Ehrlichia/Anaplasma spp. (1%), and Mycoplasma haemocanis (1%) observed. Antibodies against panleukopenia virus (67%), Toxoplasma gondii (63%), calicivirus (44%), and herpesvirus 1 (10%) were detected in cats. Feline leukemia virus antigen, feline immunodeficiency virus antibody, or coronavirus antibodies were not detected. Bartonella sp. (44%) infections were common in cats, but only one was infected with M. haemofelis. Conclusions and Clinical Importance: Despite their relative seclusion from the rest of the world, cats and dogs of Isabela were exposed to many pathogens found in mainland South America. Parasite prophylaxis, neutering, and strict enforcement of animal movement restrictions would control a majority of the diseases. In the absence of vaccination, a reservoir of susceptible animals remains vulnerable to new disease introductions.

Long-term fertility control in female cats with GonaCon™, a GnRH immunocontraceptive.

The uncontrolled reproduction of free-roaming feral cats contributes to overpopulation and associated concerns regarding their welfare and impact on public health and the environment. Nonsurgical fertility control that could be administered to feral cats in the field would be a powerful tool for cat population control. The objective was to test the efficacy and duration of activity of a single-dose GnRH immunocontraceptive vaccine (GonaCon™) on the fertility of adult female laboratory cats. Vaccinated cats (n = 15) received a single injection of vaccine containing a GnRH-KLH conjugate (200 μg) emulsified in a mycobacterial and oil adjuvant on study Day 0. Sham-treated cats (n = 5) received a single injection containing all vaccine components except the GnRH-KLH conjugate. A breeding trial started on study Day 120. Vaccinated cats had a longer time to conception (median 39.7 mo) compared to sham-treated cats (4.4 mo; P < 0.001). A total of 93% of vaccinated cats remained infertile for the first year following vaccination, whereas 73, 53, and 40% were infertile for 2, 3, and 4 y, respectively. At study termination (5 y after a single GnRH vaccine was administered), four cats (27%) remained infertile. The GnRH antibody titers declined more rapidly in short-term responding cats with < 2 y of infertility (n = 4), compared to long-term responding cats that experienced fertility control for >2 y (n = 11) (P < 0.05). Non-painful but persistent late-onset granulomatous injection site masses appeared 2 y after initial vaccination in five cats. We concluded that GnRH immunocontraception is an ideal candidate for further development for feral cat control.

Experimental Transmission of a Herpesvirus in Greek Tortoises (Testudo graeca)

An experimental transmission study aimed at fulfilling Kochs postulates for a herpesvirus-associated stomatitis-rhinitis in Mediterranean tortoises is presented. Clinical, pathologic, serologic, and molecular studies were performed linking tortoise herpesvirus with the pathogenesis of stomatitis-rhinitis. Four adult Greek tortoises received either intranasally or intramuscularly two tortoise herpesvirus isolates by primary experimental infection and secondary challenge 11 months later. After the primary experimental infection and the secondary challenge, clinical signs of illness developed, which included conjunctivitis, diphtheritic oral plaques, and oral discharge. At 4 weeks after the secondary challenge, all tortoises were humanely euthanatized and evaluated. Although neutralizing antibodies developed after the primary experimental infection, they apparently did not prevent the later development of recurrent clinical signs. Polymerase chain reaction (PCR) and reverse transcription-PCR analyses allowed sensitive characterization of the systemic distribution of the herpesvirus DNA sequences and their presence in the cranial nerves and brains of the infected tortoises. Despite the failure to recover the herpesviruses used in the transmission study, the findings support the premise that tortoise herpes-virus is a primary pathogen of Greek tortoises.

Enzyme-Linked Immunosorbent Assay for Detecting Herpesvirus Exposure in Mediterranean Tortoises (Spur-Thighed Tortoise [Testudo graeca] and Hermann's Tortoise [Testudo hermanni])

ABSTRACT An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of antibodies to a herpesvirus associated with an upper respiratory tract disease in Mediterranean tortoises [spur-thighed tortoise (Testudo graeca) and Hermanns tortoise (Testudo hermanni)]. This serodiagnostic test was validated through a hyperimmunization study. The mean of theA405 readings of the plasma samples collected at time zero of the hyperimmunization study plus three times the standard deviation was used as the cutoff for seropositivity in tortoises. ELISA results were compared to serum neutralization (SN) values for the same samples by using the McNemar test. The results obtained by SN and ELISA were not significantly different (P > 0.05). This new ELISA could be used as an important diagnostic tool for screening wild populations and private and zoo collections of Mediterranean tortoises.

Pulmonary Lesions in Experimental Ophidian Paramyxovirus Pneumonia of Aruba Island Rattlesnakes, Crotalus unicolor

Histologic and ultrastructural changes were observed in the respiratory portions of lung in five 29-40-month-old Aruba Island rattlesnakes, Crotalus unicolor, that were inoculated with an Aruba Island Rattlesnake virus (ATV) strain of ophidian paramyxovirus (OPMV) isolated from an Aruba Island rattlesnake. Lungs from one non-infected and three mock-infected Aruba Island rattlesnakes were examined also. From 4 to 22 days following intratracheal inoculation, progressive microscopic changes were seen in the lung. Initially, increased numbers of heterophils were observed in the interstitium followed by proliferation and vacuolation of epithelial cells lining faveoli. The changes appeared to progress from cranial to caudal portions of the respiratory lung following inoculation. Beginning at 4 days postinoculation, viral antigen was demonstrated in epithelial cells lining faveoli with an immunofluorescent technique using a rabbit anti-AIV polyclonal antibody. Electron microscopy revealed loss of type I cells, hyperplasia of type II cells, and interstitial infiltrates of heterophils and mononuclear cells. Viral nucleocapsid material was seen within the cytoplasm and mature virus was seen budding from cytoplasmic membranes of infected type I and type II cells from 8 to 19 days after infection. A virus consistent with AIV was isolated from lung tissues of infected rattlesnakes, thus fulfilling Kochs postulates.

CHARACTERIZATION OF PARAMYXOVIRUSES ISOLATED FROM THREE SNAKES

Multiple epizootics of pneumonia in captive snakes have been attributed to viruses which have been tentatively placed in the family Paramyxoviridae. Viruses isolated from an ill Neotropical rattlesnake (Crotalus durissus terrificus), from an Aruba Island rattlesnake (Crotalus unicolor), and from a bush viper (Atheris sp.) were propagated in Vero cells and characterized. Viral particles produced in Vero cells were pleomorphic, enveloped, and contained helical nucleocapsids. The viruses were sensitive to ether and to acidic and basic pH. Moreover, they had neuraminidase activity and were able to agglutinate erythrocytes from chicken and a variety of species of mammals. Hemagglutination was inhibited with rabbit antiserum raised against each virus. The buoyant densities of the three isolates ranged from 1.13/cm3 to 1.18/cm3, values consistent with that for an enveloped virus. The nucleic acid in the virion was determined to be RNA by [3H]uridine incorporation. Viral proteins characteristic of paramyxoviruses were immunoprecipitated from cells infected with each of the three isolates using rabbit anti-Neotropical virus serum. The morphologic appearance, physico- and biochemical properties, and cytopathologic effects of these snake viruses were consistent with those of certain members of the family Paramyxoviridae.

Immunoglobulin concentrations in feline colostrum and milk, and the requirement of colostrum for passive transfer of immunity to neonatal kittens.

The purpose of this study was to clarify whether cats have a colostral and milk phase of lactation differentiated by concentrations of immunoglobulins, and whether colostrum ingestion by newborn kittens is essential for optimal transfer of passive immunity. Milk from specific pathogen-free queens was analyzed for IgG and IgA concentrations from parturition through 6 weeks of lactation. Serum IgG and IgA concentrations from birth through 8 weeks of age were determined for colostrum-fed kittens, colostrum-deprived kittens that were fed a milk replacer, and colostrum-deprived kittens that were fostered onto queens in the milk phase of lactation. The total IgG and IgA concentrations in milk were significantly higher on the day of parturition than on day 7 of lactation, indicating cats do have a colostral phase of lactation. The predominant immunoglobulin in both colostrum and milk was IgG. The serum IgG concentrations in colostrum-deprived kittens fostered on queens in the milk phase of lactation were similar to colostrum-deprived kittens fed a milk replacer, and the concentrations were significantly lower than in colostrum-fed kittens for the first 4 weeks of life. The serum IgA concentrations in both colostrum-deprived groups were significantly lower than colostrum-fed kittens on day 2 after parturition, but were similar thereafter. Colostrum-deprived kittens fostered onto queens in the milk phase of lactation had failure of passive transfer of maternal antibodies. Protective concentrations of immunoglobulins can be restored in kittens with failure of passive transfer of immunity by parenteral administration of adult cat serum, but not by fostering on queens in mid-lactation.

Validation and use of an indirect enzyme-linked immunosorbent assay for detection of antibodies to West Nile virus in American alligators (Alligator mississippiensis) in Florida

In October 2002, West Nile virus (WNV) was identified in farmed American alligators (Alligator mississippiensis) in Florida showing clinical signs and having microscopic lesions indicative of central nervous system disease. To perform seroepidemiologic studies, an indirect enzyme-linked immunosorbent assay (ELISA) was developed to determine exposure of captive and wild alligators to WNV. To validate the test, a group of WNV-seropositive and -seronegative alligators were identified at the affected farm using hemagglutination inhibition (HAI) and the plaque reduction neutralization test (PRNT). The indirect ELISA utilized a rabbit anti-alligator immunoglobulins polyclonal antibody as the secondary antibody, and inactivated WNV-infected Vero cells were used as the coating antigen. For all samples (n=58), the results of the ELISA were consistent with the HAI and PRNT findings. Plasma was collected from 669 free-ranging alligators from 21 sites across Florida in April and October 2003. Four samples collected in April and six in October were positive for WNV antibodies using HAI, PRNT, and the indirect ELISA. This indicated that wild alligators in Florida have been exposed to WNV. These findings can be used as a baseline for future surveys.

Effects of maternally-derived antibodies on serologic responses to vaccination in kittens

The optimal vaccination protocol to induce immunity in kittens with maternal antibodies is unknown. The objective of this study was to determine the effects of maternally-derived antibody (MDA) on serologic responses to vaccination in kittens. Vaccination with a modified live virus (MLV) product was more effective than an inactivated (IA) product at inducing protective antibody titers (PAT) against feline panleukopenia virus (FPV). IA vaccination against feline herpesvirus-1 (FHV) and feline calicivirus (FCV) was more effective in the presence of low MDA than high MDA. Among kittens with low MDA, MLV vaccination against FCV was more effective than IA vaccination. A total of 15%, 44% and 4% of kittens had insufficient titers against FPV, FHV and FCV, respectively, at 17 weeks of age. Serologic response to vaccination of kittens varies based on vaccination type and MDA level. In most situations, MLV vaccination should be utilized and protocols continued beyond 14 weeks of age to optimize response by all kittens.