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.

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.

Herpesviruses are well-known infectious agents with remarkably wide host ranges. Starting in 1975 (33), several reports have documented the presence of herpesvirus-like particles in land tortoises and freshwater and marine turtles (5, 7-9, 11, 15, 17, 19-23, 27-29, 30, 38). Recent investigations have revealed an association between the presence of herpesvirus and an upper respiratory tract disease in Mediterranean tortoises [spur-thighed tortoise (Testudo graeca) and Hermann's tortoise (T. hermanni)] (5,8,9,17,20,23,(27)(28)(29)30).In tortoises with herpesvirus infection, clinical signs range from a mild conjunctivitis to a severe stomatitis-glossitis and pharyngitis. Diphtheritic plaques can be observed on the dorsal surface of the tongue and on the hard palate of infected tortoises. Frequently, a clear serous to a mucopurulent nasal discharge is present. Signs of central nervous system disease have also been reported in Mediterranean tortoises with herpesvirus infection (17).Eosinophilic intranuclear inclusions, often seen in multiple tissues, are particularly prominent in tortoises with pharyngitis and glossitis. As seen with transmission electron microscopy, inclusions consist of numerous viral particles. The morphology and morphogenesis have been used to categorize the virus as herpesvirus.A diagnosis of herpesvirus infection is often made based solely upon light or electron microscopy findings. Antemortem diagnosis can be made using biopsy specimens of oral lesions. A serum neutralization (SN) test has been developed but is limited in its application since it is only available in a few research laboratories in Europe (10). In addition, time is a limiting factor with the SN test. Ten to 14 days are required to obtain the final reading and a laborious procedure is required. An easier and faster but equally reliable serodiagnostic test is needed. In this report, we describe the development of an enzyme-linked immunosorbent assay (ELISA) that can be used to monitor the exposure to herpesvirus of free-range, private, and zoo collections of tortoises. MATERIALS AND METHODSViruses. Herpesvirus isolates HV1976 and HV4295/7R/95 were used as antigens in the ELISAs and immunoblotting. HV1976 was isolated from a captive Hermann's tortoise from the United States (Washington), while HV4295/7R/95 was isolated from a captive Hermann's tortoise in Germany during a herpesvirus outbreak in a private collection (27).Antigen preparation for ELISA. The herpesvirus isolates were grown in terrapene heart cell monolayers (TH-1; ATCC-CCL 50 Sub-line B1; American Type Culture Collection, Rockville, Md.) in T-150 plastic flasks with ventilated caps (Corning, Rochester, N.Y.) for use as ELISA antigens. The TH-1 cells were grown in Dulbecco's modified Eagle's medium F12 (Gibco BRL, Grand Island, N.Y.) with 5% fetal bovine serum (Sigma, St. Louis, Mo.), gentamicin (60 mg/liter) (Sigma), penicillin G (120,000 U/liter), streptomycin (120,000 U/liter), and amphotericin B (300 g/liter) (ABAM; Sigma). Infected cell monolayers were scraped a...

Abstract. An experimental transmission study aimed at fulfilling Koch's 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 herpesvirus is a primary pathogen of Greek tortoises.

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.