Seroprevalence of Toxoplasma gondii in American free-ranging or captive pumas (Felis concolor) and bobcats (Lynx rufus)

Toxoplasma gondii is a major zoonotic agent infecting a wide range of mammals, including wild felids. Like domestic cats, wild felids are involved in the complete infective cycle of T. gondii, as they can host in their gastrointestinal tract sexually mature parasites and shed infective oocysts in their feces. In order to evaluate the importance of this wildlife reservoir, 438 serum samples collected between 1984 and 1999 from 438 pumas (Felis concolor) and from 58 bobcats (Lynx rufus) from North America, Central America and South America were screened for antibodies to T. gondii. The overall prevalence of T. gondii antibodies was 22.4% in pumas and 51.7% in bobcats, with regional variations. Adults were more likely to be seropositive than juveniles and kittens (prevalence ratio (PR) = 2.61; confidence interval (CI) = 1.15, 4.04). In the US, pumas from the southwestern states (Arizona, California and New Mexico) were more likely to be seropositive for T. gondii ( PR = 2.61; 95% CI = 1.32-5.18 ) than pumas from the northwestern and mountain states (Colorado, Idaho, Oregon, Utah and Wyoming). Male pumas from the US were more likely to be seropositive than females (PR = 2.08; 95% CI = 1.11-3.92), whereas female pumas from Mexico, Central America and South America were more likely to be seropositive than female pumas from Canada and the US (PR = 2.49; 95% CI = 1.09-5.69). Captive pumas were also more likely to be seropositive (21.7%, 29/92) for T. gondii than free-ranging animals (19.9%, 69/346) (PR = 1.85; 95% CI = 1.06, 3.17).     

Blood parameters of clinically normal captive bobcats (Felis rufus).

Data on blood parameters were obtained from 56 (32 female, 24 male) clinically healthy, captive bobcats (Felis rufus). The animals were housed under ambient conditions at the Mississippi State University Captive Bobcat Facility. Samples were collected opportunistically between 1982 and 1995. Differences (P < or = 0.05) in blood parameters existed between sexes, among age classes (kitten, subadult, adult, and geriatric), and among stages in the reproductive cycle (breeding, parturition, recycling, and anestrus). Blood parameters for bobcats were similar to reported values for domestic cats (Felis catus). These results can be used as as reference to indicate when blood parameter aberrations exist in bobcats.     

Experimental transmission of Cytauxzoon felis from bobcats (Lynx rufus) to domestic cats (Felis domesticus)

Freshly collected blood and/or spleen homogenate from an experimentally infected Florida bobcat (Lynx rufus floridanus), which had died of feline cytauxzoonosis, was inoculated into domestic cats. All inoculated cats had clinical signs of feline cytauxzoonosis and died within 2 weeks after they were inoculated. Similar material collected from an eastern bobcat (Lynx rufus rufus) carrying an experimentally infected Cytauxzoon felis parasitemia was inoculated into domestic cats. All inoculated cats developed a parasitemia, but none developed clinical signs of disease and none died of the disease. Cats subinoculated with parasitemic cat blood also developed parasitemias and they too did not develop clinical signs of infection nor died. After carrying the blood phase of Cytauxzoon felis for various periods, the domestic cats were then challenge exposed with proven lethal Cytauxzoon inoculum of domestic cat origin. All cats died of cytauxzoonosis.     

A comparison of two field chemical immobilization techniques for bobcats (Lynx rufus)

Anesthetic protocols that allow quick induction, short processing time, and rapid reversal are necessary for researchers performing minimally invasive procedures (including morphometric measurements or attachment of radiocollars). The objective of this study was to evaluate the effectiveness of medetomidine and butorphanol as a substitute for xylazine in ketamine-based field immobilization protocols for bobcats (Lynx rufus) to reduce recovery and total field times. During 2008 and 2009, 11 bobcats were immobilized with an intramuscular combination of ketamine (10 mg/kg)-xylazine (0.75 mg/kg) (KX) or ketamine (4 mg/kg)-medetomidine (40 mcg/kg)-butorphanol (0.4 mg/kg) (KMB). Time to initial sedation, recumbency, and full anesthesia were recorded postinjection. Time to head up, sternal, standing, full recovery, and total processing times were recorded post-reversal. Throughout anesthesia, heart rate (HR), respiratory rate (RR), rectal temperature (RT), and noninvasive hemoglobin-oxygen saturation (SpO2) were recorded at 5-min intervals. The KX combination had a median time to full anesthesia of 10 min, a median recovery time of 46 min, and a median total processing time of 83 min. Alternatively, the KMB combination had a median time to full anesthesia of 21 min, a median recovery time of 18 min, and a median total processing time of 64 min. The KX protocol produced a median HR of 129 beats/min, RR of 25 breaths/min, RT of 38.3 degrees C, and SpO2 of 93%. The KMB protocol produced a median HR of 97 beats/min, RR of 33 breaths/min, RT of 38.4 degrees C, and SpO2 of 92%. Though both protocols provided safe and reliable sedation, the benefits of using medetomidine and butorphanol to lower ketamine doses and decrease processing time for brief nonsurgical sedation of bobcats in the field are presented.     

Observation of Helicobacter-like organisms in gastric mucosa of grey foxes (Urocyon cinereoargenteus) and bobcats (Lynx rufus)

Archival specimens of gastric mucosa of 10 raccoons (Procyon lotor), 9 porcupines (Erethizon dorsatum), 6 grey foxes (Urocyon cinereoargenteus), 6 bobcats (Lynx rufus), 4 skunks (Mephitis mephitis), and 3 black bears (Ursus americanus) were microscopically examined for evidence of Helicobacter-like organisms. Such organisms were seen in the specimens from the grey foxes and bobcats only. Histochemical stains (modified Steiner and carbol fuchsin methods) revealed long spiral organisms within lumina of gastric glands; however, neither gross nor microscopic lesions were observed. By electron microscopy (EM), the organisms were found to be free in the glandular lumina and were seen occasionally in the cytoplasm of gastric epithelial cells. Morphologically, 2 different phenotypes of spiral organisms were identified by EM. The organisms associated with bobcats appeared to be more tightly coiled than those seen in grey foxes. The presence of Helicobacter-like organisms in the gastric mucosa of grey foxes has not previously been described.     

Spongiform encephalopathy in a captive puma (Felis concolor).

A captive adult puma developed ataxia, a hypermetric gait and whole body tremor. The signs progressed over a period of six weeks. Histopathological examination following euthanasia demonstrated spongiform encephalopathy, gliosis and mild non-suppurative meningoencephalitis. Immunostaining with a polyclonal antiserum revealed prion protein (PrP) associated with these changes in sections of cervical spinal cord and medulla. This is the first confirmed case of a scrapie-like spongiform encephalopathy described in a non-domestic cat in the United Kingdom.     

Dermatophyte infections in free-ranging Florida panthers (Felis concolor coryi).

Three free-ranging Florida panthers (Felis concolor coryi) were diagnosed with clinical dermatophytosis; two were infected with Trichophyton mentagrophytes, and one was infected with Microsporum gypseum. Two of these panthers were juvenile males that were diagnosed with focal to focally coalescing dermatophytosis; one caused by M. gypseum and the other by T. mentagrophytes. These animals were not treated, and clinical signs resolved spontaneously over 6 mo. The third panther, an adult male from southern Florida, presented with a diffuse dermatophytosis due to T. mentagrophytes infection. Initially, the panther had alopecia, excoriations, ulcerations, and multifocal pyoderma of the head, ears, neck, rear limbs, and abdominal region that progressed to lichenification of the skin and loss of nails from two digits. When topical therapy applied in the field at 45-day intervals was ineffective in clearing the infection, the animal was placed in captivity for intensive oral therapy to prevent further development of dermal mycosis, loss of additional nails, and spread of infection to other panthers. The panther was treated orally with itraconazole (9.5 mg/ kg) in the food s.i.d. for 6 wk. After treatment, nail regrowth occurred but the multifocal areas of alopecia remained. The panther was released back into the wild after two skin biopsy cultures were negative for fungal growth. Temporary removal of a free-ranging animal of an endangered species from its habitat for systemic treatment of dermatophytosis requires consideration of factors such as age, reproductive potential, holding facilities, treatment regimen, and the potential for successful reintroduction of the animal.     

Bartonella species detection in captive, stranded and free-ranging cetaceans

We present prevalence of Bartonella spp. for multiple cohorts of wild and captive cetaceans. One hundred and six cetaceans including 86 bottlenose dolphins (71 free-ranging, 14 captive in a facility with a dolphin experiencing debility of unknown origin, 1 stranded), 11 striped dolphins, 4 harbor porpoises, 3 Risso's dolphins, 1 dwarf sperm whale and 1 pygmy sperm whale (all stranded) were sampled. Whole blood (n = 95 live animals) and tissues (n = 15 freshly dead animals) were screened by PCR (n = 106 animals), PCR of enrichment cultures (n = 50 animals), and subcultures (n = 50 animals). Bartonella spp. were detected from 17 cetaceans, including 12 by direct extraction PCR of blood or tissues, 6 by PCR of enrichment cultures, and 4 by subculture isolation. Bartonella spp. were more commonly detected from the captive (6/14, 43%) than from free-ranging (2/71, 2.8%) bottlenose dolphins, and were commonly detected from the stranded animals (9/21, 43%; 3/11 striped dolphins, 3/4 harbor porpoises, 2/3 Risso's dolphins, 1/1 pygmy sperm whale, 0/1 dwarf sperm whale, 0/1 bottlenose dolphin). Sequencing identified a Bartonella spp. most similar to B. henselae San Antonio 2 in eight cases (4 bottlenose dolphins, 2 striped dolphins, 2 harbor porpoises), B. henselae Houston 1 in three cases (2 Risso's dolphins, 1 harbor porpoise), and untyped in six cases (4 bottlenose dolphins, 1 striped dolphin, 1 pygmy sperm whale). Although disease causation has not been established, Bartonella species were detected more commonly from cetaceans that were overtly debilitated or were cohabiting in captivity with a debilitated animal than from free-ranging animals. The detection of Bartonella spp. from cetaceans may be of pathophysiological concern.     

Cytauxzoon felis infections are present in bobcats (Lynx rufus) in a region where cytauxzoonosis is not recognized in domestic cats

This study was performed to determine the prevalence of Cytauxzoon felis (C. felis) infections in bobcats (Lynx rufus) from a region where C. felis is recognized in domestic cats, North Carolina (NC), and a region where C. felis is not recognized in domestic cats, Pennsylvania (PA). Samples from NC (n=32) were obtained post-mortem via cardiac puncture from legally trapped bobcats. Samples from PA (n=70) were collected post-mortem onto Nobuto blood collecting strips by the PA Game Commission. Each sample was tested using a C. felis specific PCR assay as well as a PCR assay targeting host DNA to rule out the presence of PCR inhibitors. Three samples were excluded due to the presence of PCR inhibitors. Thirty-three percent (10/30) of the samples from NC and 7% (5/69) of the samples from PA tested positive for the presence of C. felis. The proportion of C. felis positive bobcats from NC was significantly different than that from PA (P<0.005). Despite the lower prevalence of C. felis infections in bobcats from PA this finding is unique and indicates the potential for C. felis infections in domestic cats in the northeastern USA if the appropriate tick vectors are present. Veterinary practitioners in PA should be on alert for cytauxzoonosis in domestic cats. Further studies about the epidemiology and transmission of C. felis infections among both domestic cats and bobcats are needed.     

Embryo transfer and embryonic capsules in the bobcat (Lynx rufus)

Bobcats (Lynx rufus) (n=22) were used to test a surgical embryo transfer protocol for wild felines. Five blastocysts were collected 8-14 days post-initial copulation (PIC). Translucent capsule-like structures were recovered at 12 days PIC and are the first report of such a structure in a felid. Endometrial fibrosis was observed in one cat but, in general, post-surgical fibrosis of the uterus did not seem to impede ova or embryo transport. One embryo underwent cryopreservation and this embryo plus two other transferrable embryos were placed in recipient cats during the course of the study. No pregnancies were maintained; but one non-cryopreserved embryo was detected by ultrasound examination at 2 weeks post-transfer. This study provides valuable groundwork for future studies and warrants optimism for continued research in this area.     

Distribution and prevalence of Cytauxzoon felis in bobcats (Lynx rufus), the natural reservoir, and other wild felids in thirteen states

Cytauxzoon felis, a protozoan parasite of wild and domestic felids, is the causative agent of cytauxzoonosis in domestic and some exotic felids in the United States. The bobcat (Lynx rufus) is the natural reservoir for this parasite, but other felids such as Florida panthers (Puma concolor coryii) and domestic cats may maintain long-term parasitemias and serve as reservoirs. Experimentally, two tick species, Dermacentor variabilis and Amblyomma americanum, have demonstrated the ability to transmit C. felis. These two tick species have overlapping distributions throughout much of the southeastern United States. The objective of the current study was to determine the distribution and prevalence of C. felis in free-ranging bobcat populations from 13 states including California, Colorado, Florida, Georgia, Kansas, Kentucky, Missouri, North Carolina, North Dakota, Ohio, Oklahoma, South Carolina, and West Virginia. These states were selected because of differential vector presence; D. variabilis is present in each of these states except for the region of Colorado sampled and A. americanum is currently known to be present only in a subset of these states. Blood or spleen samples from 696 bobcats were tested for C. felis infection by a polymerase chain reaction (PCR) assay which targeted the first ribosomal internal transcribed spacer region (ITS-1). Significantly higher prevalences of C. felis were detected from Missouri (79%, n=39), North Carolina (63%, n=8), Oklahoma (60%, n=20), South Carolina (57%, n=7), Kentucky (55%, n=74), Florida (44%, n=45), and Kansas (27%, n=41) compared with Georgia (9%, n=159), North Dakota (2.4%, n=124), Ohio (0%, n=19), West Virginia (0%, n=37), California (0%, n=26), and Colorado (0%, n=67). In addition to bobcats, seven cougars (Puma concolor) from Georgia, Louisiana, and North Dakota and one serval (Leptailurus serval) from Louisiana were tested for C. felis. Only one cougar from Louisiana was PCR positive, which represents the first report of an infected cougar outside of the Florida panther population. These data also indicate that C. felis is present in North Dakota where infection has not been reported in domestic cats. Based on a nonparametric analysis, prevalence rates were significantly higher in states where there are established populations of A. americanum, which supports recent data on the experimental transmission of C. felis by A. americanum and the fact that domestic cat clinical cases are temporally associated with A. americanum activity. Collectively, these data confirm that bobcats are a common reservoir for C. felis and that A. americanum is likely an epidemiologically important vector.     

Diagnosis and treatment of primary hyperparathyroidism in a bobcat (Lynx rufus)

An 18-yr-old male bobcat (Lynx rufus) presented with chronic moderate weight loss and acute onset of anorexia and lethargy. Hypercalcemia and azotemia were present on the serum chemistry panel. Abdominal ultrasound revealed hyperechoic renal cortices, but no evidence of neoplasia. Ionized calcium and 25-hydroxyvitamin D were mildly elevated, intact parathyroid hormone was severely elevated, and parathormone-related protein was undetected, suggesting primary hyperparathyroidism with possible renal dysfunction. Azotemia lessened in severity following diuresis, but hypercalcemia persisted; thus primary hyperparathyroidism was considered the most probable differential diagnosis. A second ultrasound including the cervical region revealed a solitary intraparenchymal left thyroid nodule. The nodule was surgically excised; histopathology confirmed a parathyroid adenoma. Although primary hyperparathyroidism was suspected, diagnosis was not achieved from serum chemistry values alone. This case emphasizes the importance of diagnostic imaging and histopathology in the investigation of persistently abnormal laboratory values.     

Seroprevalence of retrovirus in North American captive macropodidae

Laboratory records of serology results from captive macropodidae sampled between 1997 and 2005 were reviewed to assess the seroprevalence of retrovirus exposure. Serum samples from 269 individuals (136 males, 133 females) representing 10 species of macropods housed in 31 North American captive collections were analyzed for retrovirus antibody using an indirect immunofluorescent assay. The prevalence of positive antibody titers comparing male versus female, between species, between age groups, and among animals with identified parentage was examined by nonparametric statistical analyses. Median age of animals at time of sample collection was 36 mo (range 2-201 mo). Total percentage seropositive was 20.4%. Serum antibody was detected in 31 of 47 (66.0%) tammar wallaby (Macropus eugenii), nine of 24 (37.5%) yellow-footed rock wallaby (Petrogale xanthopus), four of 11 (36.4%) swamp wallaby (Wallabia bicolor), 10 of 80 (12.5%) red-necked wallaby (Macropus rufogriseus), and one of 54 (1.9%) parma wallaby (Macropus parma). No individuals of western gray kangaroo (n=3) (Macropus fuliginosus), eastern gray kangaroo (n=19) (Macropus giganteus), common wallaroo (n=6) (Macropus robustus), red kangaroo (n=11) (Macropus rufus), or Matschie's tree kangaroo (n=14) (Dendrolagus matschiei) were positive for retrovirus antibody. These results demonstrate that five species of captive macropods have a history of exposure to retrovirus, with the highest percentage seropositive and highest statistical correlation in M. eugenii (pair-wise Fisher's exact test, alpha = 0.05). Additionally, one wild-caught M. eugenii was confirmed seropositive during quarantine period, indicating that retrovirus exposure may exist in wild populations.     

Ectoparasites of free-ranging pumas and jaguars in the Paraguayan Chaco

Ectoparasites were collected from seven puma (Puma concolor) and seven jaguar (Panthera onca) live-captures (each representing six different animals) in the Paraguayan Chaco from 2002 to 2004. The same five species of ectoparasites were recovered from both host species: the flea, Pulex simulans (total on both hosts combined=30 male, 49 female), and the ticks, Amblyomma cajennense (1 male, 4 female, 46 nymphs, 241 larvae), Amblyomma parvum (42 male, 25 female), Amblyomma tigrinum (1 male, 7 female, 34 larvae) and Amblyomma triste (4 male). There were no statistical differences between the prevalences (percent of hosts infested) for any of these ectoparasite species between the two host species, and only P. simulans showed a significantly higher mean intensity (mean no. of each ectoparasite species per infested host) on one of the host species (jaguar). Clearly, these two large carnivores share similar ectoparasite faunas in the Paraguayan Chaco. However, both carnivores occupy large geographical distributions in the New World and this study, combined with the few previous studies, suggests that their ectoparasite faunas differ slightly in different parts of their respective ranges.     

Virological Survey in free-ranging wildcats (Felis silvestris) and feral domestic cats in Portugal

To determine the presence of viral pathogens in natural areas a survey was conducted on an opportunistic sample of fifty eight wild (Felis silvestris silvestris) and feral cats (F. s. catus). The biological materials included serum, lung tissue extract and stool. Feline leukemia virus p27 antigen was detected in 13/50 serum/lung tissue extract samples (26%), canine distemper virus antibodies were detected in 2/26 serum/lung tissue extract samples (7.7%), feline coronavirus RNA was present in 6/29 stool samples (20.7%) and feline parvovirus DNA in 2/29 stool samples (6.9%). Canine distemper virus RNA was not detected. Feline immunodeficiency virus and feline coronavirus antibodies were not detected. Evidence of exposure to feline leukemia virus, canine distemper virus, feline coronavirus and feline parvovirus was found in wild and feral cats raising the importance of performing a comprehensive survey to correctly evaluate the potential threat of infectious diseases to endangered species, namely to the wildcat and to the Iberian lynx, which is meant to be reintroduced after 2012 in Portugal.     

Seroprevalence of Neospora caninum and Toxoplasma gondii in captive and free-ranging nondomestic felids in the United States.

Seven serum samples of 101 samples from nondomestic, captive and free-ranging felids from the United States were indirect fluorescent antibody positive for antibodies to Neospora caninum, whereas 44 samples were positive for antibodies to T. gondii. Although none of the captive animals displayed clinical signs of disease, nondomestic felids in the United States have been exposed to, and are likely infected with, N. caninum and T. gondii. This may have serious implications for zoological gardens exhibiting susceptible animals, such as kangaroos, close to felids.     

Development and validation of puma (Felis concolor) cytokine and lentivirus real-time PCR detection systems

Studies of immune correlates of disease outcome associate humoral immune response mediated by T-helper 2 cytokines (IL-4, IL-10) with more virulent disease relative to a cell-mediated response driven by T-helper 1 cytokines (IL-2, IFN-gamma), particularly in viral and other intra-cellular infections. Specifically, the kinetics of both human immunodeficiency virus (HIV) and feline immunodeficiency virus (FIV) infection are closely associated with Type 1 versus Type 2 cytokine profiles. Puma (Felis concolor) lentivirus (PLV) is closely related to FIV, but based on phylogenetic and clinical studies, is more ancient and less pathogenic. The aims of this study were to validate feline real-time PCR primer/probe systems for puma cytokines and PLV as sensitive, quantitative assays for use in investigations of PLV pathogenicity. We demonstrate that primer/probe systems for IL-4, IL-10, IFN-gamma, TNF-alpha, GAPDH, and the pol region of PLV-1695 amplify puma cytokines and PLV-1695 with high amplification efficiency and sensitivity. Detection of PLV-1695 provirus in experimentally inoculated domestic cats proved to be of equivalent sensitivity, specificity, and positive and negative predictive value to co-culture of one million peripheral blood mononuclear cells (PBMC). Evaluation of cytokine induction during naturally occurring PLV infection will allow insight into mechanisms of host control associated with apathogenic infection. In addition, determination of viral loads during different stages of PLV infection or in different tissues from domestic cats or pumas will further elucidate capacity of these viruses to replicate and establish infection.     

Clinical Parelaphostrongylus tenuis infection in two captive American bison (Bison bison)

CASE DESCRIPTION: 2 juvenile (17 and 19 months of age) male American bison (Bison bison) were examined because of acute bilateral hind limb weakness and ataxia; 1 animal also had urinary incontinence. TREATMENT AND OUTCOME: Given the clinical signs and rapid deterioration in the condition of these 2 animals, obtaining a definitive diagnosis was considered essential to minimizing the risk of disease in the remaining bison herd and among other animals at the facility. Therefore, both affected animals were euthanized. At necropsy, no gross abnormalities were seen. Histologic examination of sections of the brains from both animals revealed mild to moderate multifocal aggregates of eosinophils and mononuclear cells in perivascular regions of the meninges and gray matter of the cerebrum, cerebellum, and brainstem. Systematic examination of multiple sections of brain and spinal cord revealed evidence of nematode sections and aberrant parasite migration. CLINICAL RELEVANCE: Findings suggested that CNS migration of Parelaphostrongylus tenuis in American bison may cause clinical signs. These findings have implications for the management of captive bison and free-ranging bison sharing ranges with white-tailed deer (Odocoileus virginianus), the definitive host, and elk (Cervus elaphus canadensis).