Anaplasma sp. and hemoplasma infection in leopard cats (Prionailurus bengalensis euptilurus) from Korea

This study examined the occurrence of Anaplasma spp. and hemoplasma infection in leopard cats, Prionailurus bengalensis euptilurus, in Korea. Twenty-nine biological samples were tested by molecular analysis. Two (6.9%) and eight (27.6%) tested specimens were positive for Anaplasma bovis and hemoplasma infection, respectively. Based on our results, Anaplasma/Ehrlichia spp. and hemoplasma are regularly infecting leopard cat populations of Korea. Considering their endangered status, regular monitoring of infection by arthropod-borne pathogens known to cause clinical symptoms in feline hosts such as Anaplasma/Ehrlichia spp. and hemoplasma would be crucial as part of ongoing conservation efforts.     

Genetic and biological characterization of feline foamy virus isolated from a leopard cat (Prionailurus bengalensis) in Vietnam

Foamy viruses have been isolated from various mammals and show long-term co-speciation with their hosts. However, the frequent inter-species transmission of feline foamy viruses (FFVs) from domestic cats to wild cats across genera has been reported. Because infectious molecular clones of FFVs derived from wild cats have not been available, whether there are specific characteristics enabling FFVs to adapt to the new host species is still unknown. Here, we obtained the complete genome sequences of two FFV isolates (strains NV138 and SV201) from leopard cats (Prionailurus bengalensis) in Vietnam and constructed an infectious molecular clone, named pLC960, from strain NV138. The growth kinetics of the virus derived from pLC960 were comparable to those of other FFVs derived from domestic cats. Phylogenetic analysis revealed that these two FFVs from leopard cats are clustered in the same clade as FFVs from domestic cats in Vietnam. Comparisons of the amino acid sequences of Env and Bet proteins showed more than 97% identity among samples and no specific amino acid substitutions between FFVs from domestic cats and ones from leopard cats. These results indicate the absence of genetic constraint of FFVs for interspecies transmission from domestic cats to leopard cats.     

Characterization of blood cells in the Leopard Cat (Prionailurus bengalensis)

Background: The Leopard Cat (Prionailurus bengalensis) is the most frequently encountered wild cat in most of Southeast Asia. Limited hematologic investigation exists for this species. Objectives: The objectives of this study were to assess routine hematologic measurements and parameters and characterize the morphology, cytochemical staining, and ultrastructural features of blood cells in Leopard Cats. Methods: Blood samples were collected from 12 adult healthy captive Leopard Cats (7 males and 5 females). Complete blood counts were performed using an automated hematology analyzer and manual differential counts. Cytochemical staining (Sudan black B [SBB], peroxidase [PO], periodic acid‐Schiff [PAS], α‐naphthyl acetate esterase [ANAE], and β‐glucuronidase [BG]) and scanning and transmission electron microscopy were performed using standard methods. Results: Median (range) hematologic results were as follows: PCV 0.46 L/L (0.30–0.55 L/L), hemoglobin 136.5 g/L (100–183 g/L), WBC 9.0 × 10⁹/L (6.9–15.2 × 10⁹/L), band neutrophils 0.07 × 10⁹/L (0–0.30 × 10⁹/L), segmented neutrophils 2.9 × 10⁹/L (1.2–6.34 × 10⁹/L), lymphocytes 5.3 × 10⁹/L (2.7–8.1 × 10⁹/L), eosinophils 0.14 × 10⁹/L (0–0.73 × 10⁹/L), basophils 0/L (0–0.22 × 10⁹/L), and monocytes 0.08 × 10⁹/L (0–0.30 × 10⁹/L). Neutrophils stained strongly positive for SBB, PO, and PAS; lymphocytes had fine granular positivity for ANAE and BG; monocytes were weakly positive for ANAE and BG; and basophils were strongly positive for BG. Ultrastructurally, eosinophils contained many large rod‐shaped granules with prominent crystalloid core structures, ribosomes, and mitochondria. Basophils contained many round to oval specific granules with homogeneous contents. Low number of basophils contained a few small vacuoles that usually were not detected by light microscopy. Conclusion: These findings will facilitate interpretation of hematologic results for future investigative and diagnostic studies of this species.     

Hematology and serum biochemistry of free-ranging nutria (Myocastor coypus)

Information on reference blood values in the literature is lacking for many wild rodents. In this study, comprehensive reference intervals (RIs) for a wide range of analytes from 101 healthy free-ranging nutria were determined. Animals were captured in Buenos Aires, Argentina (37degrees 50'S, 57 degrees 34'W), and southward (38 degrees 60'S, 58 degrees 23'W), encompassing major biotopes of agricultural pampas with dunes and grassland steppes on the east coast. Traps were set at locations with high-density nutria populations (i.e., those areas that showed signs of movement, territorial marking, or feeding activities). Although the small sample size limits the interpretation of these findings, RIs were determined by a robust method using the central 95th percentile. In nutria, the RI range varied greatly for the leukocyte differentials, with mature neutrophils: 3,907-5,544/mmicrol for females and 3,744-5,900/microl for males; band neutrophils: 0-10/ll for females and 3-18/microl for males; lymphocytes: 4,213 5,940/microl for both sexes combined; monocytes: 165-402/microl for both sexes combined; eosinophils: 13-91/microl for females and 108-165/microl for males; and basophils: 0-87/microl for both sexes combined. Platelet concentration was 543-727 x 10(9)/L for both sexes combined. There was also a wide RI range for biochemistry values for some enzymes, such as alkaline phosphatase: 200-399 IU/L for both sexes combined; cholinesterase: 762-1,407 IU/L for females and 763-1,284 IU/L for males; creatine kinase: 182-552 IU/L for females and 162-451 IU/L for males; amylase: 853-1,865 IU/L for females and 779-1,293 IU/L for males; and glucose concentration 120.2-180.6 mg/dl for both sexes combined. Conversely, there was not a wide pooled RI range for calcium: 7.0-11.2 mg/dl; phosphorous: 6.1-9.3 mg/dl; sodium: 133.0-159.0 mEq/L; potassium: 3.0-8.2 mEq/L; chloride: 101.4-143.0 mEq/L; and urea: 11.3-36.8 mg/dl. The red blood cell indices had a narrow range, with mean corpuscular volume: 84.0 -102.5 fl and mean corpuscular hemoglobin concentration: 18.2-28.8 g/dl, and which was most likely due to strict physiologic controls. The results from this study were similar to those previously reported for farmed nutria.     

Sperm cryopreservation in the Far‐Eastern wildcat (Prionailurus bengalensis euptilurus)

The Far‐Eastern wildcat (Prionailurus bengalensis euptilurus) is a rare and poorly investigated nondomestic felid species. An attempt of freezing and cryopreserving Far‐Eastern wildcat spermatozoa in CaniPlus Freeze (CPF) medium is reported. Sperm was collected by electroejaculation from five adult Far‐Eastern wildcat captive‐born males. Epididymal spermatozoa from five adult randomly bred domestic cat males were used as a reference. The viability of frozen–thawed spermatozoa evaluated by double staining with SYBR Green I and PI followed by the subsequent confocal laser scanning microscopy (CLSM) was 38.2% ± 3.0% for the domestic cat and 38.0% ± 10.2% for the Far‐Eastern wildcat. The motility of frozen–thawed spermatozoa was 30.8% ± 9.8% for the domestic cat and 33.7% ± 15.1% for the Far‐Eastern wildcat. Sperm morphology was assessed by light microscopy. The total percentage of normal spermatozoa after freezing and thawing was 51.9 ± 5.9 for the domestic cat and 55.0% ± 6.4% for the Far‐Eastern wildcat. Defects of flagella were the most frequently observed abnormalities in both species (32.2% ± 4.8% and 30.8% ± 4.4% of all reported anomalies for the domestic cat and Far‐Eastern wildcat, respectively). Domestic cat epididymal and Far‐Eastern ejaculatory spermatozoa fertilized in vitro‐matured oocytes of the domestic cat (30.0% ± 5.5% and 35.5% ± 15.0%, respectively). Taken together, these results suggest that the freezing of Far‐Eastern wildcat spermatozoa with CPF medium is a suitable method for Felidae cryopreservation.     

Hematologic and serum biochemistry values in two species of free-ranging porcupines (Coendou prehensilis, Coendou melanurus) in French Guiana.

Individuals of two porcupine species (Coendou prehensilis and Coendou melanurus) were translocated during the flooding of a forest at a hydroelectric dam site in French Guiana. Blood samples were collected for 11 mo to determine hematologic and serum chemistry reference mean values and ranges and to look for blood parasites. Male C. prehensilis had significantly higher hemoglobin levels, packed cell volumes, and creatinine and potassium values than did females of the same species. Coendou prehensilis had significantly lower amylase levels but higher creatinine, calcium, lactate dehydrogenase, and gamma glutamyl transferase levels than C. melanurus.     

Cholesterol values in free-ranging gorillas (Gorilla gorilla gorilla and Gorilla beringei) and Bornean orangutans (Pongo pygmaeus).

Cholesterol concentrations in captive gorillas and orangutans vary widely within species and average approximately 244 mg/dl for gorillas and 169 mg/dl for orangutans as published previously. The International Species Inventory System reports higher concentrations of 275 and 199 mg/dl for gorillas and orangutans, respectively. It is unknown whether these values were typical, influenced by captive management, or both. To answer this question, banked serum samples from free-ranging mountain gorillas (Gorilla beringei), western lowland gorillas (Gorilla gorilla gorilla), and Bornean orangutans (Pongo pygmaeus) were analyzed for total cholesterol, triglyceride, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol concentrations. Mountain gorillas did not differ significantly from free-ranging western lowland gorillas in cholesterol, triglyceride, high-density lipoprotein cholesterol, or low-density lipoprotein cholesterol concentrations, indicating mountain gorilla values could be a model for western lowland gorillas. Captive gorilla total cholesterol and low-density lipoprotein cholesterol concentrations were significantly higher (P < 0.05) than in free-ranging groups. Triglyceride concentrations for captive gorillas were significantly higher (P < 0.05) than the male mountain and western lowland gorillas, but they were not significantly different from the female mountain gorillas. Captive orangutan total cholesterol concentrations were only higher (P < 0.05) than the free-ranging female orangutans, whereas captive orangutan low-density lipoprotein cholesterol concentrations were significantly higher (P < 0.05) than both free-ranging male and female orangutans. Calculated and measured low-density lipoprotein cholesterol concentrations were compared for all free-ranging animals and were significantly different (P < 0.05) for all groups, indicating Friedewald's equation for calculating low-density lipoprotein cholesterol is not appropriate for use with nonfasted apes. The higher total cholesterol and low-density lipoprotein cholesterol concentrations in captive apes may predispose them to cardiovascular disease and might be attributed to diets, limited energy expenditure, and genetics.     

Molecular characterization of Hepatozoon species in a Leopard Cat (Prionailurus bengalensis) from Thailand

Abstract: Hepatozoon gamonts were observed by light microscopy in neutrophils of a male, wild‐caught Leopard Cat. Complete blood counts at presentation and 6 months later were unremarkable. Serologic tests were negative for both FIV and FeLV. A partial sequence of the 18S rRNA gene from the Hepatozoon found in the cat indicated that, compared with all species examined, the protozoan had the closest relationship (99.2% sequence similarity) with the Hepatozoon of the water python (Stegonotus cucullatus). The cat was clinically healthy at last report. Although Hepatozoon has been found in another wild cat in Thailand, this is the first report in a Leopard Cat. The pathogenicity of Hepatozoon in these cats remains uncertain.     

Hematology and serum biochemistry values of trapped, healthy, free-ranging rock hyraxes (Procavia capensis) and their association with age, sex, and gestational status

BACKGROUND: The rock hyrax (Procavia capensis) is an herbivore prevalent from South Africa to Turkey, and a most common zoo animal. Although many studies of hyrax diseases and physiology are available, clinicopathologic data are limited. OBJECTIVES: The purpose of this study was to establish comprehensive hematologic and biochemical reference intervals for trapped, apparently healthy, free-ranging rock hyraxes using modern laboratory methods and to assess differences related to sex, gestation, and age. METHODS: Blood samples were obtained from 27 healthy, free-ranging hyraxes under anesthesia. Gender, body weight, and gestational status were recorded. Hematologic (n = 25) and serum biochemical (n = 22) analyses were performed using standard automated methodology. Data for male vs female, adult vs juvenile, and pregnant vs nonpregnant female hyraxes were compared using the Mann-Whitney U-test. Associations between variables were assessed using Pearson's or Spearman rank correlation tests. RESULTS: Significant age- and sex-related, but not gestation-related differences were observed in several variables. Serum alkaline phosphatase activity and phosphorus concentration were significantly higher in juveniles compared with adults. A unique type of monocyte comprised 1-3% of leukocytes in 4 hyraxes. Markedly high serum creatine kinase (CK) activity was observed in most hyraxes. CONCLUSIONS: The large number of animals and the availability of sex, age, and gestational data in this study will be useful to zoo and wildlife veterinarians working with rock hyraxes. High serum concentrations of betahydroxybutyric acid in the rock hyrax, compared with dogs, cats, and ruminants, may be related to its unique digestive system. High CK activity may have been the result of a capture myopathy-like syndrome. The unique monocytes in hyraxes resemble those of elephants and are a novel finding in this species.     

Effects of in vitro hemolysis on serum biochemistry values of the bottlenose dolphin (Tursiops truncatus).

The effects of in vitro hemolysis on 23 biochemical analytes were assessed in sera from 14 clinically healthy Atlantic bottlenose dolphins (Tursiops truncatus). Each serum sample was divided into three portions for analysis: 1) nonhemolyzed control; 2) moderate hemolysis, simulated by adding hemolyzed serum to a final concentration of approximately 150 mg/dl Hb; and 3) severe hemolysis, simulated by adding hemolyzed serum to a final concentration of approximately 500 mg/dl Hb. Moderate hemolysis resulted in statistically significant increases in the mean values of iron, lactate dehydrogenase, potassium, and uric acid and a decrease in creatinine (P < 0.001). Severe hemolysis resulted in statistically significant changes in the mean values of the above analytes in addition to the following increases: alanine aminotransferase, calcium, and serum globulins (P < 0.001) and albumin and total protein (P < 0.01). Total bilirubin and gamma glutamyl transferase levels were lower in the severely hemolyzed sample (P < 0.001). Differences in mean values for alkaline phosphatase between nonhemolyzed and hemolyzed serum were not significant but did show a downward trend in the hemolyzed sera. The presence and severity of hemolysis must be considered in the interpretation of the serum chemistry values.     

Hematology and clinical chemistry values of free-ranging basilisk lizards (Basiliscus plumifrons) in Costa Rica

Twenty-three lizards were captured for this study, both males and females (12 males, 10 females, 1 undetermined), with a large range in body weights (40-286 g) appeared to be healthy based on activity level, physical examinations, and body condition scores. Heparinized blood samples from 20 free-ranging basilisk lizards (Basiliscus plumifrons) in Costa Rica were used for determining complete blood cell counts, plasma, and heparinized whole blood biochemical analysis. This information will serve as baseline reference data for future health assessment studies of free-ranging and captive basilisk lizards, as well as epidemiologic, conservation, and captive-breeding studies. A point-of-care analyzer was useful for this field study, and clinical chemistry values from heparinized whole blood samples were similar to values from plasma, which indicates that separation of plasma may not be necessary to process blood samples on site in remote areas. To the authors' knowledge, this is the first report of hematologic and plasma biochemical data from free-ranging B. plumnifrons.     

Cross‐sectional observational survey of serum biochemistry values in a population of 69 adult female alpacas (Vicugna pacos) in South Australia

Blood samples were collected from 69 ‘healthy’ female alpacas aged ≥12 months from 11 properties in South Australia. The 10–90 percentile ranges of the 16/19 analytes measured in this sample population were within the published ranges of four healthy alpaca populations from other geographic locations. Marginal exceptions were glutamate dehydrogenase and bicarbonate. Potassium was notably elevated, probably because of haemolysis of some samples. The sample size was insufficient to provide the appropriate statistical power to define diagnostic references ranges according to international standards. The health status of the sample population of alpacas was presumptive based on a physical examination.     

Serum thyroxine (T4) and triiodothyronine (T3) uptake values in normal adult cats as determined by radioimmunoassay

Radioimmunoassay procedures employing highly specific antibodies for either serum T4 or T3 were utilized to determine T4 concentrations and T3 uptake values in normal cats. Mean values for serum T4 and for T3 uptake were 30.9 ng/ml +/- 19 ng/ml and 59.67% +/- 12.2%, respectively.     

Transitional cell carcinoma in fishing cats (Prionailurus viverrinus): pathology and expression of cyclooxygenase-1, -2, and p53

A high prevalence of urinary bladder transitional-cell carcinoma (TCC) has been noted in captive fishing cats (Prionailurus viverrinus). Of the 91 adult deaths between 1995 and 2004, 12 (13%) were attributed to TCC. To help elucidate mechanisms of carcinogenesis, archival sections of urinary bladder from 14 fishing cats were examined histologically and immunohistochemically for p53, cyclooxygenase (COX)-1, and COX-2 expression. Ten cats had TCC, and 4 were unaffected. The average age at death was 10.8 years in affected individuals and 10.5 years in unaffected individuals. There was no sex predilection. Fishing cat TCCs were characterized histologically as papillary and infiltrating (n = 6), nonpapillary and infiltrating (n = 3), or carcinoma in situ (n = 1). Glandular and squamous metaplasia, necrosis, and lymphatic invasion were prominent histologic features. Two individuals had documented metastasis. p53 nuclear immunolabeling was detected in 4/10 (40%) TCCs. In two cases, immunolabeling was limited to less than 10% of the neoplastic cellular population and was comparable to staining of normal fishing cat bladder. Therefore, p53 gene mutation did not appear to be an essential component of TCC carcinogenesis in examined fishing cats. COX-1 immunohistochemistry was negative in all cases. All TCCs had some degree of COX-2 cytoplasmic immunolabeling, which was exclusively within the invasive portions of the neoplasms. Papillary portions were uniformly negative. COX-2 overexpression was a prominent feature in the majority of the examined fishing cat TCCs, suggesting that COX-2-mediated mechanisms of carcinogenesis are important in this species and that COX-inhibiting drugs may be of therapeutic benefit.     

Coagulation values in normal ferrets (Mustela putorius furo) using selected methods and reagents

Background: Accurate determination of commonly measured coagulation values would be useful in the diagnosis and management of coagulopathies in domestic ferrets (Mustela putorius furo). We are unaware of reports of coagulation times in this species. Objectives: The purpose of this study was to determine reference values for prothrombin time (PT), activated partial thromboplastin time (PTT), fibrinogen concentration, and antithrombin (AT) activity in ferrets using selected methods and reagents. Methods: Blood samples obtained from 18 clinically healthy ferrets were anticoagulated with 0.129 M sodium citrate in a ratio of 9 parts blood to 1 part anticoagulant. Plasma was collected and stored at -70 degrees C until analysis. PT and PTT were measured with a fibrometer and with an ACL 3000 automated system. PTT was measured with and without the addition of ellagic acid. Fibrinogen was assayed by a turbidimetric method. AT activity was determined using a chromogenic assay and pooled ferret plasma (100% activity). Differences in methods and reagents were evaluated using paired t tests. Results: PT was significantly longer using the fibrometer (12.3+/-0.3, 11.6-12.7 seconds) compared with the ACL (10.9+/-0.3, 10.6-11.6 seconds) (P<.01). PTT was not significantly different with the fibrometer (18.7+/-0.9, 17.5-21.1 seconds) vs the ACL (18.1+/-1.1, 16.5-20.5 seconds), but was significantly longer on both analyzers when ellagic acid was added (fibrometer 20.4+/-0.8, 18.9-22.3 seconds; ACL 20.0+/-1.0, 18.6-22.1 seconds) (P<.01). Fibrinogen concentration was 107.4+/-19.8 mg/dL (90.0-163.5 mg/dL), and AT activity was 96%+/-12.7% (69.3-115.3%). Conclusion: These coagulation results for healthy ferrets will be useful in the evaluation of ferrets with coagulopathies, provided similar reagents and methods are used.     

Blood values in free-ranging nesting leatherback sea turtles (Dermochelys coriacea) on the coast of the Republic of Gabon.

Leatherback sea turtles (Dermochelys coriacea) are the most endangered of the seven species of sea turtles. The health status of leatherbacks is largely unknown, although the number of nesting females recorded throughout the world has decreased precipitously in the last few decades. Central African beaches may provide one of the last strongholds for nesting leatherback females. In the region, oil extraction and incidental capture pose significant threats to the health of the population. Physical examinations, hematology, plasma biochemistry, plasma corticosterone concentration, plasma protein electrophoresis, plasma vitamin concentrations, and toxicological parameters were evaluated in nesting female leatherbacks in the Republic of Gabon. The general clinical condition of the 35 turtles examined in this study was rated as good. The blood value results for a subset of these turtles are presented and compared to published results from other sea turtles. To the authors' knowledge, these are the first published baseline hematology, plasma biochemistry, and plasma protein electrophoresis values from clinically healthy nesting leatherback turtles.     

Elusive cats in our backyards: persistence of the North Chinese leopard (Panthera pardus japonensis) in a human-dominated landscape in central China

The North Chinese leopard (Panthera pardus japonensis), the least-known big cat, disappeared in most historical range for decades, following the development of modern civilization. Unfortunately, we have scarce knowledge about the status of this big cat so far, apart from anecdotal reports. In this study, we investigated density, distribution, and habitat use of the leopard, the apex predator, in a complex forest landscape in the Loess Plateau. We used a camera-trapping network to obtain population estimates for leopards over 2 years through spatially explicit capture–recapture models. Our results, based on maximum likelihood and Bayesian/MCMC methods, reveal that the largest wild population of the leopard was found widely distributed in remnant forests in central Loess plateau. The population is increasing in our study area, and the density of leopards (1.70 (SE = 0.48) − 2.40 (SE = 0.67)/100 km²) is higher than other areas of China. According to the analysis of 2 seasonal occupancy models, prey species drive partially the leopard habitat use, predicting that the big cat thrives from the recovery of prey community. However, human disturbances, especially oil wells, seem to have negative impacts on the habitat use of leopards. Specifically, it is necessary to have joint efforts by the government and researchers to improve human disturbances management and prey species population density, as well as strengthen the investment in research on the North Chinese leopard, which could all further strengthen protection ability and ensure the long-term survival of this species.