Wild Bobcats Research Articles

Semen collection and characterization of normative reproductive traits in free-ranging ocelots (Leopardus pardalis) and bobcats (Lynx rufus) in southern Texas

Decreased genetic diversity and possible inbreeding depression have recently been documented in the last wild ocelot (Leopardus pardalis) population in the United States. One consequence of inbreeding depression in felids may be reduced semen quality which can adversely affect reproductive potential. Detailed assessments of reproductive parameters in wild individuals and populations can be conducted using assisted reproductive technologies, such as semen collection and analysis. For most felid species, semen has traditionally been collected via electroejaculation (EEJ22Electroejaculation); however, an alternative method has been developed using alpha-2 agonist drugs to induce direct sperm release into the urethra, allowing collection by catheterization without requiring specialized equipment. The goal of this study was to characterize normative reproductive traits in free-ranging ocelots and co-occurring bobcats (Lynx rufus) in southern Texas and assess the effectiveness of urethral catheterization (UC33Urethral Catheterization) for semen recovery in both species. For semen collection, free-ranging cats were live-captured and anesthetized using intramuscular ketamine and medetomidine/dexmedetomidine (alpha-2 agonist) with UC conducted 20–40 minutes post-induction. In ocelots only, EEJ was subsequently performed if UC failed to recover a viable sample. Semen collection was attempted in 31 felids (n=9 ocelots; n=22 bobcats), with sperm recovery by UC in seven of nine ocelots (78 %) and 14 of 22 bobcats (66 %), and by EEJ in four of five ocelots (80 %). For ocelots, the percentage of primary morphologic abnormalities was higher (p<0.001) for UC (47.75 ± 6.7; mean ± SEM) compared to EEJ (9 ± 2.7) but percent normal morphology (MORPH) did not differ between UC and EEJ (p=0.218). In wild ocelots, seminal parameters appeared lower relative to historical values reported for zoo-managed ocelots, possibly related to reduced heterozygosity. In wild bobcats, seminal traits were inferior to those of ocelots but similar to reports for other zoo-managed Lynx species. In conclusion, detailed male reproductive traits have been characterized for the first time in wild, free-ranging ocelots and bobcats in southern Texas. Although UC allowed semen recovery for assessment of seminal traits in both species, EEJ produced higher quality samples in ocelots when applied after UC while also mitigating the adverse impact of urine contamination observed frequently with both collection methods.

HIGH PREVALENCE OF CYTAUXZOON FELIS IN BOBCATS (LYNX RUFUS) ACROSS OKLAHOMA AND OCCURRENCE IN WEST TEXAS, USA.

Cytauxzoonosis is a fatal tick-borne disease in domestic cats caused by infection with the apicomplexan Cytauxzoon felis. Bobcats are the natural wild-vertebrate reservoirs for C. felis, and infections are typically subclinical and chronic in this species. The present study was done to determine the prevalence and geographic distribution of C. felis infection in wild bobcats from Oklahoma and the occurrence in northwestern Texas. Tongue samples from 360 bobcats were collected from 53 counties in Oklahoma and 13 samples from three counties in Texas. For DNA extracted from each tongue sample, a probe-based droplet digital PCR assay was performed targeting the C. felis mitochondrial gene cytochrome c oxidase subunit III (cox3). Prevalence of C. felis infection was calculated for each county sampled, and data from individual counties were combined according to geographic regions and compared using chi-square tests. Overall prevalence of C. felis in bobcats from Oklahoma was 80.0% (95% confidence interval [CI], 75.6-83.8). The prevalence of infection was >90% for bobcats from central, northeastern, south-central, and southeastern regions of Oklahoma, but <68% for bobcats from northwestern and southwestern regions. Bobcats from central counties in Oklahoma were 25.693 times more likely to be infected with C. felis compared to all other bobcats sampled from the state. Higher prevalence estimates of C. felis in bobcats appeared to be in counties where known tick vectors are most common. Occurrence of C. felis in bobcats from northwestern Texas was 30.8% (95% CI, 12.4%-58.0%) based on 13 samples. Results of this study support the utilization of bobcats as sentinel animals to identify geographic areas with risk of C. felis infection to domestic cats.

Field immobilization of Desert Bobcats (Lynx rufus) using a Tiletamine-Zolazepam anesthetic combination

Chemical immobilization is used to minimize stress and pain while safely handling wild animals under field conditions. Eight wild bobcats (Lynx rufus) were immobilized for radio collar fittings between March 2005 and March 2007, using a commercial tiletamine hydrochloride-zolazepam hydrochloride combination (Zoletil® Virbac, Carros, France). Bobcats were captured using soft catch spring traps (Victor No. 3) in the Sonoran Desert of Baja California Sur, Mexico. A mean (±SD) intramuscular total dose of 44.3 ± 2.9 mg (6.0 mg kg–1) produced an induction time of 9 ± 4.9 min, and a duration of cataleptic anesthesia of 46.5 ±11.4 min. No adverse reactions were observed in animals during handling, and telemetry monitoring indicated that all individuals were alive for at least one year after the procedure. The results of this study indicate that use of tiletamine-zolazepam administered at 6 mg/kg dose is suitable for short-term chemical immobilization of bobcats in the wild.

High prevalence of Lynx rufus gammaherpesvirus 1 in wild Vermont bobcats.

Gammaherpesviruses (GHVs) are host specific DNA viruses that infect a large range of mammalian species. These viruses preferentially target host lymphocyte cell populations and infection may lead to morbidity or mortality in immunocompromised, co-infected, or non-adapted hosts. In this study, we tested for the presence of Lynx rufus gammaherpesvirus 1 (LruGHV1) in a northeastern United States population of wild bobcats (L. rufus). We estimated prevalence of infection and viral load in infected individuals using quantitative real-time PCR analysis of spleen DNA from 64 Vermont bobcats. We observed an overall prevalence of 64% using this methodology. Bobcat age was significantly positively associated with GHV infection status, and we noted a trend for higher viral loads in young animals, but prevalence and viral load were similar in male and female bobcats. A single LruGHV1 variant was identified from the sequencing of the viral glycoprotein B gene of Vermont bobcats. This gene sequence was 100% similar to that reported in Florida bobcats and slightly variant from other isolates identified in the Western USA. Our work suggests broad geographic distribution and high prevalence of LruGHV1 in bobcat populations across the United States with infection attributes that suggest horizontal transmission of the agent. Geographic differences in viral genotype may reflect historical migration and expansion events among bobcat populations.

Bobcats (Lynx rufus) are natural definitive host of Besnoitia darlingi

Bovine besnoitiosis, caused by Besnoitia besnoiti, is an economically important disease of cattle in many countries but its transmission remains a mystery. Wild felids are suspected to be its definitive hosts. The domestic cat (Felis catus) is known experimental definitive host for Besnoitia species of rodents. Here, we report for Besnoitia darlingi the first identification of a natural definitive host, the bobcat (Lynx rufus). Oocysts resembling Toxoplasma gondii (unsporulated; 10.9±0.8×12.1±0.2μm; n=5) were detected microscopically in the feces of two of 25 free ranging wild bobcats from Mississippi, USA. After detailed investigation, we identified these oocysts as B. darlingi and not T. gondii. The IFN-γ gene knockout (KO) mice fed oocysts from bobcats died of acute besnoitiosis and tachyzoites were found in their tissues. Oocysts were also mildly pathogenic to outbred Swiss Webster mice (SW) (Mus musculus). The SW mice fed oocysts became ill but generally survived and developed characteristic thick-walled Besnoitia tissue cysts in their tongue and heart muscles and brains. Two laboratory-raised domestic cats (Felis catus) excreted B. darlingi oocysts after ingesting murine tissues infected with bobcat-derived oocysts. The parasite was successfully cultivated in African green monkey kidney fibroblast cells (CV-1 cell line) seeded with infected murine tissue homogenate. The multilocus PCR-DNA sequencing (18S rDNA, 28S rDNA, and ITS-1) from culture-derived tachyzoites confirmed the parasite as B. darlingi. Our results suggest that bobcats may be an important link in the sylvatic cycle of Besnoitia species and bioassay or molecular tests are needed to differentiate Toxoplasma gondii-like oocysts in feces of felids, both domestic and wild cats.

Seroprevalence, isolation and co-infection of multiple Toxoplasma gondii strains in individual bobcats (Lynx rufus) from Mississippi, USA

Toxoplasma gondii causes lifelong chronic infection in both feline definitive hosts and intermediate hosts. Multiple exposures to the parasite are likely to occur in nature due to high environmental contamination. Here, we present data of high seroprevalence and multiple T. gondii strain co-infections in individual bobcats (Lynx rufus). Unfrozen samples (blood, heart, tongue and faeces) were collected from 35 free ranging wild bobcats from Mississippi, USA. Toxoplasma gondii antibodies were detected in serum by the modified agglutination test (1:≥200) in all 35 bobcats. Hearts from all bobcats were bioassayed in mice and viable T. gondii was isolated from 21; these strains were further propagated in cell culture. Additionally, DNA was extracted from digests of tongues and hearts of all 35 bobcats; T. gondii DNA was detected in tissues of all 35 bobcats. Genetic characterisation of DNA from cell culture-derived isolates was performed by multiplex PCR using 10 PCR-RFLP markers. Results showed that ToxoDB genotype #5 predominated (in 18 isolates) with a few other types (#24 in two isolates, and #2 in one isolate). PCR-DNA sequencing at two polymorphic markers, GRA6 and GRA7, detected multiple recombinant strains co-infecting the tissues of bobcats; most possessing Type II alleles at GRA7 versus Type X (HG-12) alleles at GRA6. Our results suggest that individual bobcats have been exposed to more than one parasite strain during their life time.

Metal accumulation in bobcats in the Southeastern USA.

Bobcats (Lynx rufus) are wide-ranging mammals found throughout the continental USA. As carnivores near the top of their food chain, bobcats would seem to be a useful bioindicator of metal pollution in terrestrial environments. However, there is very limited research on bobcats in toxicology studies. Here, we offer the first analysis of metal (copper, selenium, silver, and zinc) contaminants in the livers of wild bobcats. Liver tissues from 120 adult bobcats (i.e., estimated to be ≥1year old) were collected from 2003 to 2006 at four sites in Georgia and Florida, USA that experienced relatively similar levels of human disturbance. We found no differences in metal concentrations between males and females. At two of the sites sampled over three consecutive years, there was substantial year-to-year variation in the concentrations of Cu, Se, and Zn. We also documented some variation between sites, but only between sites sampled in different years, which may reflect additional temporal, rather than spatial, variation. Concentrations of Cu and Ag were significantly positively correlated with one another, as were concentrations of Se and Zn. Contrary to expectation, there were no significant relationships between body weight and metal concentrations. Finally, comparison with results from previous metal toxicology studies of nine-banded armadillos (Dasypus novemcinctus) and Virginia opossums (Didelphis virgianus), collected from the same sites during the same years, showed differential patterns of accumulation across species, suggesting that ecological lifestyle is an important influence on metal accumulation. This study provides reference levels of metal contaminants in the liver of bobcats as well as insight into metal accumulation in a top level carnivore.

Antibody Detection and Molecular Characterization of Toxoplasma gondii from Bobcats (Lynx rufus), Domestic Cats (Felis catus), and Wildlife from Minnesota, USA.

Little is known of the epidemiology of toxoplasmosis in Minnesota. Here, we evaluated Toxoplasma gondii infection in 50 wild bobcats (Lynx rufus) and 75 other animals on/near 10 cattle farms. Antibodies to T.gondii were assayed in serum samples or tissue fluids by the modified agglutination test (MAT, cut-off 1:25). Twenty nine of 50 bobcats and 15 of 41 wildlife trapped on the vicinity of 10 farms and nine of 16 adult domestic cats (Felis catus) and six of 14 domestic dogs resident on farms were seropositive. Toxoplasma gondii oocysts were not found in feces of any felid. Tissues of all seropositive wild animals trapped on the farm were bioassayed in mice and viable T.gondii was isolated from two badgers (Taxidea taxus), two raccoons (Procyon lotor), one coyote (Canis latrans), and one opossum (Didelphis virginiana). All six T.gondii isolates were further propagated in cell culture. Multi-locus PCR-RFLP genotyping using 10 markers (SAG1, SAG2 (5'-3'SAG2, and alt.SAG2), SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico), and DNA from cell culture derived tachyzoites revealed three genotypes; #5 ToxoDataBase (1 coyote, 1 raccoon), #1 (1 badger, 1 raccoon, 1 opossum), and #2 (1 badger). This is the first report of T.gondii prevalence in domestic cats and in bobcats from Minnesota, and the first isolation of viable T.gondii from badger.

Dental Eruption in Bobcats

We documented tooth eruption in 9 hand-raised bobcat (Felis rufus) kittens. Tooth lengths were measured to the nearest mm. Deciduous dentition began to erupt at 11-14 days of age and was completed by 9 weeks of age. Permanent dentition began to erupt at 16-19 weeks of age and was completed by 34 weeks of age. The sequence and timing of deciduous and permanent dentition will aid in estimating parturition and copulation dates from bobcat kittens in the wild. J. WILDL. MANAGE. 52(3):515-517 Information on the dental eruption patterns of known-age bobcats has not been published. Data on sex ratios, age structures, and age-specific reproductive rates have been collected and analyzed by states to assess the status of resident bobcat populations (Gluesing 1982). These data have been compared to data collected and analyzed in a similar manner from prior studies (Gluesing et al. 1986). Therefore, researchers have compared 1 unknown to another when estimating age structure and periods of reproductive activity. Estimates of breeding and parturition dates of the bobcat have been based on dentition-eruption schedules and growth rates of the domestic cat. Crowe (1975) developed a dentition eruption schedule for bobcats using the tooth eruption schedule of the domestic cat presented by McClure et al. (1973:211). Crowe's (1975) schedule has been used to age wild bobcat kittens (Blankenship 1979). The purpose of our study was to determine eruption schedules for deciduous and permanent teeth from knownage bobcat kittens. An accurate knowledge of the tooth eruption schedules of bobcat kittens will aid in elucidating parturition and conception dates of wild bobcats. This manuscript is dedicated to E. A. Gluesing, who initiated the study and whose efforts and guidance made this study possible. We thank J. R. Jackson, V. M. Woshner, and others who helped raise the kittens. We also thank B. D. Leopold, E. P. Hill, and R. M. Kaminski for their help in editing this manuscript, and K. M. Vandevelde for typing it. Financial support was p ovided by Mississippi Federal Aid in Wildlife Restoration Project W-48-30 and the Mississippi Agricultural and Forestry Experiment Station.

Food Intake and Assimilation by Bobcats under Laboratory Conditions

Food Intake and Assimilation by Bobcats under Laboratory Conditions

Odd Items in Bobcat Stomachs

These brief remarks on stomach contents of wild bobcats, Lynx rufus, are occasioned by my recent unearthing of eight poorly preserved specimens, each a stomach and small intestine, cryptically tagged “Cat Stomachs.” Two of them were further labeled, “Wild Cat—shot ahead of dogs in Montmorency County, Mich. Dec. 12, 1932. Reported to be eating snowshoe hares.” Measurements of the animals were appended. No data accompanied the others, but it seems probable that all eight represent approximately the same locality and date. Three of the specimens were practically empty (moist contents less than 10 cc); contents of one measured 160 cc, of another 358 cc, and of the remaining three from 55 to 75 cc. The contents of all were, for the most part, unrecognizable. However, one stomach and one intestine contained tightly packed masses of the nematode Physaloptera praeputialis?, kindly identified by William D. Lindquist. This genus has previously been reported by Hamilton and Hunter (Jour. Wildl. Mgt., 3: 99–103, 1939) as common in Vermont bobcats.