Characterization of Inflammasomes and Their Regulation in the Red Fox.

BACKGROUND: Inflammasomes recognize endogenous and exogenous danger signals, and subsequently induce the secretion of IL-1ß. Studying inflammasomes in the red fox (Vulpes vulpes) is crucial for wildlife veterinary medicine, as it can help control inflammatory diseases in foxes. METHODS: We investigated the activation and intracellular mechanisms of three inflammasomes (NLRP3, AIM2, and NLRC4) in fox peripheral blood mononuclear cells (PBMCs), using established triggers and inhibitors derived from humans and mice. RESULTS: Fox PBMCs exhibited normal activation and induction of IL-1ß secretion in response to representative inflammasome triggers (ATP and nigericin for NLRP3, dsDNA for AIM2, flagellin for NLRC4). Additionally, PBMCs showed normal IL-1ß secretion when inoculated with inflammasome-activating bacteria. In inhibitors of the inflammasome signaling pathway, fox inflammasome activation was compared with mouse inflammasomes. MCC950, a selective NLRP3 inhibitor, suppressed the secretion of dsDNA- and flagellin-mediated IL-1ß in foxes, unlike mice. CONCLUSIONS: These findings suggest that NLRP3 may have a common role in dsDNA- and flagellin-mediated inflammasome activation in the red fox. It implies that this fox inflammasome biology can be applied to the treatment of inflammasome-mediated diseases in the red fox.

Treatment of two Asiatic black bears (Ursus thibetanus) with severe injuries and their subsequent release into the wild: a case report.

BACKGROUND: The rehabilitation of injured wildlife and their subsequent release into the wild is a humane act as well as important in wildlife conservation. However, little is known about the animals' fate after release. Therefore, to address these uncertainties, it is essential to adequately describe how the injured animals were treated and managed before releasing into the wild; moreover, post-release monitoring should also be performed. Herein, we document for the first time the process of rescue, surgery, and rehabilitation of severely injured Asiatic black bears (Ursus thibetanus; endangered species in South Korea) and their fate after returning to the wild. CASE PRESENTATION: A six-year-old female (bear-01) and a three-year-old male (bear-02) bears were injured by an illegal snare and collision with a bus, respectively. Bear-01 had broad muscle necrosis and ruptures from the snared ankle on the right thoracic limb, with myiasis, and elbow disarticulation was performed. In bear-02, a non-reducible comminuted fracture of the left humerus was confirmed radiologically, and the operation was performed by using dual plate fixation with hydroxyapatite and recombinant human bone morphogenetic protein-2. The bear-01 and -02 were completely healed approximately 30 and 60 days after surgery, respectively. After that, they underwent rehabilitation for 8 and 25 days, respectively, in an outdoor enclosure similar to their natural habitat. Bear-01 and -02 were released into the wild after 45 and 99 days after surgery, respectively, and their mean daily movement distance during the first 30 days after releasing was 2.9 ± 2.1 and 1.3 ± 1.6 km, respectively. The annual mean 95% Kernel home-range size of bear-01 and bear-02 was 265.8 and 486.9 km2, respectively. They hibernated every winter, gained weight, gave birth to cubs (bear-01), were not found to have any abnormalities in the veterinary tests, and were not involved in any conflicts with humans after returning to the wild. CONCLUSIONS: Bears without one leg or those with dual plates could adapt well in their natural habitat, which shows that our surgical and post-operative treatments were effective. Additionally, minimizing human contact and observing/evaluating behavior during the rehabilitation is essential in reducing human-bear conflicts after release.

A Clonorchiasis Case of a Leopard Cat, Prionailurus bengalensis euptilurus, Diagnosed by Ultrasonography and Egg Detection in Republic of Korea.

A clonorchiasis case in a captive leopard cat, Prionailurus bengalensis euptilurus, was confirmed by ultrasonographic findings and egg morphologies found in the bile juice sample in the Korea. The leopard cat was introduced from the wild habitat of Gyeongsangnam-do, to Cheongju Zoo in Cheongju-si, Chungcheongbuk-do, Korea in August 2014. Physical examinations were basically performed for quarantine and check-up health. The cat was comparatively good in health except anorexia. The cyst-like bile duct dilation and the increased echogenicity of gall bladder wall and hepatic parenchyma were observed by ultrasonography. Ultrasound-guided needle biopsy was conducted for collecting bile juice and the specimens were observed under light microscope. The numerous small trematode eggs were detected in the bile juice sample of the light microscopy. The eggs were 25-33 (28±3) µm by 18-22 (20±1) µm in size and showed typical characteristics of Clonorchis sinensis egg, i.e., a dominantly developed operculum, shoulder rim and dust-like wrinkles in surface. To treat the liver fluke infection, 20 mg/kg of praziquantel was orally administered only once to the case. Follow-up studies including fecal examinations were conducted during 2 years after treatment. But no more eggs were detected from the case. In the present study, we described the first clonorchiasis case of leopard cat, which was confirmed by ultrasonographic findings and egg morphologies from the bile juice sample in Korea.

Effectiveness of urethral catheterization under ultrasound guidance for semen collection from Asiatic black bears (Ursus thibetanus).

The Asiatic black bear (ABB; Ursus thibetanus ussuricus) is a globally endangered species, and measures to help increase their population are necessary. For the successful restoration of this species, artificial breeding as well as conservation translocation are considered important. The aims of the present study were to evaluate the feasibility and effectiveness of urethral catheterization (UC), which is effectively used in feline species, for semen collection from ABBs and establish the optimal protocol for semen collection via this technique. Seven clinically healthy, adult male ABBs (age, 6-13 years; weight, 130-180 kg) housed at the Species Restoration Technology Institute, Korea were included in this study. All study procedures were performed during the breeding season (June to August) over 3 consecutive years. Semen samples were collected once or three times from all bears by ultrasound-guided UC or electroejaculation (EE) under general anesthesia, and their characteristics, including sperm motility, were evaluated. The day of semen collection was defined as Day 0. The semen collected by the UC method was stored at 4 °C, and sperm motility was evaluated at the same time every day for 16 days. The successful collection rates for the UC and EE methods were 92.3% and 53.8%, respectively. The sperm concentration (4718.9 ±â€¯1526.1 vs. 185.0 ±â€¯34.2 × 106/ml), total sperm count (1196.6 ±â€¯955.5 vs. 100.9 ±â€¯70.0 × 106), sperm motility score (4.39 ±â€¯0.78 vs. 3.00 ±â€¯1.73), viability (98.2 ±â€¯2.3 vs. 82.7 ±â€¯19.6), and the proportion of spermatozoa with intact acrosomes (92.2% ±â€¯9.3% vs. 75.6% ±â€¯10.6%) were higher with the UC method than with the EE method, whereas the proportion of spermatozoa with an abnormal morphology (23.1% ±â€¯4.6% vs. 45.6% ±â€¯19.5%) was lower with the former than with the latter. Over the course of cool storage, there was an overall decrease in the total motility, progressive motility, and viability, although viability was >50% until Day 10. These findings suggest that ultrasound-guided UC is a useful and feasible tool for the collection of high-quality semen from ABBs. The collected semen remains viable for up to 10 days, with high sperm motility maintained for up to 7 days, when stored at 4 °C.

FLUOXETINE THERAPY TO DECREASE STEREOTYPIC BEHAVIOR IN THE ASIATIC BLACK BEAR (URSUS THIBETANUS).

The present study evaluated the effects of fluoxetine on decreasing stereotypic behaviors in Asiatic black bears (ABB). Four captive ABB exhibiting stereotypic behaviors were administered fluoxetine (0.25-1 mg/ kg orally every 24 hr) for 91 days. The animals were monitored twice weekly for 213 days (April to October) using a scan sampling method (preadministration, 30 day; administration, 91 days; postadministration, 92 days). Five blood samples were collected from the jugular vein of each bear for serum chemistry analysis (one during preadministration; two each during administration and postadministration periods). Fluoxetine was safe and effective in decreasing stereotypies, but it may be difficult to decrease long-established stereotypies in ABB using fluoxetine alone. To decrease stereotypies in ABB effectively, it is necessary to identify underlying problems that contribute to stereotypic behavior and apply comprehensive interventions that can include fluoxetine treatment.

Infections of Two Isthmiophora Species (Digenea: Echinostomatidae) in Wild Mammals from Republic of Korea with Their Morphological Descriptions.

Isthmiophora hortensis (Digenea: Echinostomatidae) is a dominant echinostome in animal reservoir hosts and humans in the Republic of Korea (Korea). We intended to investigate the infection status with this echinostome species in the several species of wild animals and describe the morphological characteristics in the faunistic view point. A total of 175 animal carcasses belonging to 3 families, i.e., Canidae, Felidae and Mustelidae, were collected from the southern regions of Korea from March 2010 to July 2017. Isthmiophora spp. worms were recovered from the small intestines of each animal under a stereomicroscope after washing of intestinal contents. Isthmiophora hortensis was recovered from 4 species of wild carnivores, i.e., Nyctereutes procyonoides (3/107: 2.8%), Mustela sibirica (11/31: 35.5%), Meles lucurus (2/3: 33.3%) and Martes flavigula (1/2: 50%). The other 3 carnivores comprising stray dogs, cat and leopard cat were negative for I. hortensis infection (0/2, 0/10 and 0/12, respectively). Specimens obtained from the Lutra lutra (6/8: 75%) were identified as a distinct species, I. inermis, by morphological comparison. Isthmiophora inermis has thinner body, elongate testes and different anterior limits of vitelline fields. Detailed morphological descriptions and comparisons with the morphological characteristics are provided. Conclusively, it was confirmed for the first time that 3 species of mustelid mammals, i.e., M. sibirica, M. lucurus, and M. flavigula, are to be the new definitive hosts of I. hortensis in Korea. Additionally, I. inermis is to be newly added in the Korean echinostome fauna.

Blood Chemistry Reference Values for Free-Ranging Asiatic Black Bears ( Ursus thibetanus) by Season, Age, and Sex.

Physiological characteristics, such as blood chemistry values, are valuable for evaluating the health of the animals. To our knowledge, these values have never been reported for the free-ranging Asiatic black bear ( Ursus thibetanus; ABB). Thus, 28 blood chemistry values from 50 free-ranging ABBs captured in Jirisan National Park, Republic of Korea, from 2005 to 2016 were evaluated. The aim of this study was to establish blood chemistry reference values for the free-ranging ABBs during both the hibernating and nonhibernating seasons. During hibernation, mean values of creatinine (CRE), total cholesterol, total protein (TP), albumin (ALB), triglycerides, and magnesium were significantly higher than those during nonhibernation; however, mean values of blood urea nitrogen, urea nitrogen to creatinine (U/C) ratio, inorganic phosphorous (IP), aspartate aminotransferase, alanine aminotransferase, gamma-glutamyl transferase, lactate dehydrogenase (LDH), high-density lipoprotein cholesterol, and alkaline phosphatase (ALP) were significantly lower. Age differences (young vs. adult) were found in IP, LDH, TP, and ALB values during hibernation and in the U/C ratio, calcium, IP, ALP, creatine kinase myocardial band, CRE, total bilirubin, and uric acid values during nonhibernation. However, there were no sex differences (male vs. female).

Prediction of arterial blood gas values from venous blood gas values in Asiatic black bears (Ursus thibetanus) anesthetized with intramuscular medetomidine and zolazepam-tiletamine.

The objective of this study was to measure differences between arterial and venous blood gas parameters and to evaluate whether arterial blood gas values can be estimated from venous blood in Asiatic black bears (ABBs). Twelve healthy captive ABBs (8 males and 4 females; 8-16 years; 76.8-220 kg) were included in this study. The bears were immobilized with medetomidine and zolazepam-tiletamine using a dart gun. Arterial and venous samples were collected simultaneously at 5 and 35 min after recumbency (5- and 35-min points). Partial pressure of oxygen (PO2), partial pressure of carbon dioxide (PCO2), pH, bicarbonate (HCO3-), total carbon dioxide (TCO2), oxygen saturation of hemoglobin (SO2) and base excess (BEecf) were analyzed using a portable blood gas analyzer. There was no marked difference in measured and calculated variables over time in both venous and arterial blood except for PO2. However, arterial PO2, SO2 and pH were significantly higher and arterial PCO2, TCO2 and HCO3- were lower than those of venous samples at both 5- and 35-min points. In the regression analysis to estimate arterial values from venous values, PCO2, TCO2, HCO3-, BEecf and pH significantly showed over 0.45 in coefficient of determination value (R2), and there were little differences between actual and predicted arterial values. Although there were limits in venous gas values replaced those of arterial blood, if we could not get the arterial samples, the regression formulas for arterial values from venous blood in this study would be useful clinically, except for PO2 and SO2.

Immobilization of Asiatic Black Bears ( Ursus thibetanus ) with Medetomidine-Zolazepam-Tiletamine in South Korea.

The Asiatic black bear ( Ursus thibetanus ; ABB) is a globally endangered species for which a restoration program has been ongoing in South Korea since 2001. However, there is little information on immobilization protocols for ABBs. We evaluated the use of medetomidine-zolazepam-tiletamine for their immobilization. During 2005-13, we anesthetized 60 ABBs (32 males, 28 females; 7 mo to 12 yr old) with medetomidine 0.03-0.045 mg/kg and zolazepam-tiletamine 1.54-2.3 mg/kg; reversal of anesthesia was done with atipamezole 0.15-0.225 mg/kg administered intravenously alone or intravenously and intramuscularly (50:50). Mean (and SD) for physiologic collected for 373 immobilizations of at least 60 min were: time to sedation, 7.8 (5.4) min; anesthesia induction time, 13.7 (8.1) min; complete recovery time, 14.8 (12.4) min; respiratory rate, 14 (7) breaths/min; heart rate, 51 (16) beats/min; rectal temperature, 37.3 (1.3) C; and hemoglobin oxygen saturation, 88% (6%). Few cardiopulmonary side effects occurred during immobilization and adequate depth of anesthesia was maintained for >60 min without need for supplementation. The dosage and drug combination used was effective for immobilization of ABBs with minimal adverse effects on vital signs and can be recommended in most clinical applications.

Cardiorespiratory effects of isoflurane in Asiatic black bears (Ursus thibetanus) anesthetized with intramuscular medetomidine and zolazepam/tiletamine.

The objective of this study was to determine the dose-dependent effects of isoflurane on various cardiovascular parameters and the stable range of isoflurane concentrations in Asiatic black bears (Ursus thibetanus). Seven Asiatic black bears were intramuscularly injected with medetomidine, zolazepam and tiletamine (MZT) to induce anesthesia, and anesthesia was maintained by administering isoflurane in 100% oxygen (4 l/min) without mechanical ventilation. Several cardiovascular parameters were measured at five end-tidal isoflurane concentrations (0.5, 1.0, 1.5, 2.0, and 2.5%). Blood was collected from the femoral artery before administration of isoflurane and after each administration for immediate blood gas analysis. Isoflurane produced dose-dependent increases in heart rate, respiratory rate, minute volume, end-tidal carbon dioxide (CO2) partial pressure and the partial pressure of arterial CO2, and dose-dependent decreases in non-invasive blood pressure and tidal volume. Rectal temperature, oxygenation and acid-base balance were unaffected by isoflurane. All parameters in this study were in a clinically acceptable range at all times. The data show that the combination of MZT and isoflurane is suitable for general anesthesia in Asiatic black bears with spontaneous breathing during prolonged procedures. End-tidal isoflurane concentrations of 0.5 to 2.5% can be used in Asiatic black bears without adverse side effects.