The impact of vatinoxan on medetomidine-ketamine-midazolam immobilization in Patagonian maras (Dolichotis patagonum).

OBJECTIVE: To compare cardiovascular and ventilatory effects, immobilization quality and effects on tissue perfusion of a medetomidine-ketamine-midazolam combination with or without vatinoxan (MK-467), a peripherally acting α2-adrenoceptor antagonist. STUDY DESIGN: Randomized, blinded, crossover study. ANIMALS: A group of nine healthy Patagonian maras (Dolichotis patagonum). METHODS: Maras were immobilized twice with: 1) medetomidine hydrochloride (0.1 mg kg-1) + ketamine (5 mg kg-1) + midazolam (0.1 mg kg-1) (MKM) + saline or 2) MKM + vatinoxan hydrochloride (0.8 mg kg-1), administered intramuscularly. Drugs were mixed in the same syringe. At 20, 30 and 40 minutes after injection, invasive blood pressure, heart rate, respiration rate, end-tidal CO2, haemoglobin oxygen saturation, and muscle oxygenation were measured, arteriovenous oxygen content difference was calculated. Muscle tone, jaw tone, spontaneous blinking and palpebral reflex were evaluated. Times to initial effect, recumbency, initial arousal and control of the head were recorded. Paired t test, Wilcoxon matched-pairs signed rank test and analysis of variance were used to compare protocols; (p < 0.05). RESULTS: Vatinoxan significantly reduced systolic (p = 0.0002), mean (MAP; p < 0.0001) and diastolic (p < 0.0001) arterial blood pressures between 20 and 40 minutes. MAPs at 30 minutes (mean ± standard deviation) with MKM and MKM + vatinoxan were 105 ± 12 and 71 ± 14 mmHg, respectively. Without vatinoxan, four animals were hypertensive (MAP > 120 mmHg), whereas with vatinoxan, four animals were hypotensive (MAP < 60 mmHg). Muscle and jaw tone were significantly more frequently present with MKM (both p = 0.039). Other measurements did not significantly differ between protocols. CONCLUSIONS AND CLINICAL RELEVANCE: In Patagonian maras, vatinoxan attenuated the increase in blood pressure induced by medetomidine. Muscle and jaw tone were more frequently present with MKM, indicating that quality of immobilization with vatinoxan was more profound.

On the interplay between hypothermia and reproduction in a high arctic ungulate.

For free-ranging animals living in seasonal environments, hypometabolism (lowered metabolic rate) and hypothermia (lowered body temperature) can be effective physiological strategies to conserve energy when forage resources are low. To what extent such strategies are adopted by large mammals living under extreme conditions, as those encountered in the high Arctic, is largely unknown, especially for species where the gestation period overlaps with the period of lowest resource availability (i.e. winter). Here we investigated for the first time the level to which high arctic muskoxen (Ovibos moschatus) adopt hypothermia and tested the hypothesis that individual plasticity in the use of hypothermia depends on reproductive status. We measured core body temperature over most of the gestation period in both free-ranging muskox females in Greenland and captive female muskoxen in Alaska. We found divergent overwintering strategies according to reproductive status, where pregnant females maintained stable body temperatures during winter, while non-pregnant females exhibited a temporary decrease in their winter body temperature. These results show that muskox females use hypothermia during periods of resource scarcity, but also that the use of this strategy may be limited to non-reproducing females. Our findings suggest a trade-off between metabolically-driven energy conservation during winter and sustaining foetal growth, which may also apply to other large herbivores living in highly seasonal environments elsewhere.

Can acute stress be fatal? A systematic cross-disciplinary review.

In this review it is discussed if acute stress can be fatal. The review is based on literature searches on PubMed, PsycINFO as well as Web of Science. Literature concerning the conditions excited delirium syndrome (ExDS), malignant catatonia, takotsubo cardiomyopathy (TCM), and capture myopathy (CM) is reviewed and compared. The aim of the article is to identify and discuss a possible fatalness as well as a common pathophysiology behind these conditions. This includes a deregulated autonomic nervous system, neurocardiac reasons for myocardial damage, and rhabdomyolysis. We conclude that these conditions could be different manifestations of the same pathophysiological phenomenon. In addition, we suggest that it is possible to die from acute stress, but that it requires a prior sensitization, as seen in cocaine abusers and certain psychiatric patients, to render individuals disposed to an extreme autonomic nerve reaction. Lay summary This article compares different conditions in humans and in other animals, where it appears as if the human or animal dies with no other reason than being submitted to an extreme condition of mental stress. The conditions examined via a literature search are excited delirium syndrome, malignant catatonia and takotsubo cardiomyopathy in humans, and a capture myopathy in different mammals. The article theoretically suggests that one can die solely from acute stress, but that different forms sensitization probably goes ahead of such a fatal stress reaction. E.g. in cocaine addicts, some psychiatric patients, and in wild animals formerly subjected to stress an extreme sympathetic stress response might be immediately fatal. The article also theorizes that excited delirium syndrome, malignant catatonia, and capture myopathy could be more severe and acute variants of the temporary condition seen in takotsubo patients, also known as patients with broken heart syndrome.

IMMOBILIZING MUSKOX ( OVIBOS MOSCHATUS) UNDER HIGH ARCTIC CONDITIONS.

Immobilizing and handling large, free-ranging animals without proper facilities in harsh environmental conditions poses significant challenges. During two field expeditions, a total of 29 female muskoxen ( Ovibos moschatus) were immobilized in Northeast Greenland (74°N, 20°E). Fixed doses of immobilizing drugs were used regardless of animal size: 2 mg etorphine, 30 mg xylazine, 0.3 mg medetomidine, and 40 mg ketamine. Physiologic and behavioral monitoring was performed during the second expedition on 15 female muskoxen. The observed heart rates were 35-58 beats/min and respiratory rates were 25-30 breaths/min. Mean arterial pressures measured using oscillometry ranged between 117-142 mmHg. Pulse oximeter readings ranged from 91-98% with oxygen supplementation, nasal end-tidal carbon dioxide values were 24-42 mmHg, and rectal temperature ranged from 38.9-39.6°C. Induction time was 6-8 min, recovery time 2-6 min after reversal, and duration of anesthesia was 50-100 min. This anesthetic regime thus provided reliable immobilization with minimal pathophysiologic alterations.

EVALUATION OF TWO ENZYME-LINKED IMMUNOSORBENT ASSAYS MEASURING PREGNANCY-ASSOCIATED GLYCOPROTEINS IN THE BLOOD OF MUSKOXEN ( OVIBOS MOSCHATUS).

Pregnancy-associated glycoproteins (PAGs) are expressed by the ruminal placenta, making their detection in blood an accurate indicator of pregnancy. This study aimed to evaluate two commercially available PAG enzyme-linked immunosorbent assays (ELISAs) in muskoxen ( Ovibos moschatus). The two tests are based on the same principles; however, one is evaluated photometrically and the other visually. Sixteen samples covering all trimesters of pregnancy, and 16 nonpregnant samples were included to evaluate test performance. Both tests reliably detected pregnancy. The photometric ELISA showed a sensitivity and specificity of 94% and 100%, respectively. Although the visual ELISA depends on somewhat subjective interpretations, it came up with a sensitivity of 81% and a specificity of 100%, and might thus provide a useful in-house tool when limited laboratory equipment is available. Analysis of additional samples showed consistent results during pregnancy and circulating PAGs for at least 18 days postpartum.

A Comparative Study on the Faecal Bacterial Community and Potential Zoonotic Bacteria of Muskoxen (Ovibos moschatus) in Northeast Greenland, Northwest Greenland and Norway.

Muskoxen (Ovibos moschatus) are ruminants adapted to a high-fibre diet. There is increasing interest in the role that gut microbes play in the digestion and utilization of these specialized diets but only limited data available on the gut microbiome of high-Arctic animals. In this study, we metabarcoded the 16S rRNA region of faecal samples from muskoxen of Northeast Greenland, Northwest Greenland and Norway, and quantified the effects of physiological and temporal factors on bacterial composition. We found significant effects of body mass, year of sampling and location on the gut bacterial communities of North East Greenland muskoxen. These effects were however dwarfed by the effects of location, emphasizing the importance of the local ecology on the gut bacterial community. Habitat alterations and rising temperatures may therefore have a considerable impact on muskoxen health and reproductive success. Moreover, muskoxen are hunted and consumed in Greenland, Canada and Alaska; therefore, this study also screened for potential zoonoses of food safety interest. A total of 13 potentially zoonotic genera were identified, including the genera Erysipelothrix and Yersinia implicated in recent mass die-offs of the muskoxen themselves.

Immune function in arctic mammals: Natural killer (NK) cell-like activity in polar bear, muskox and reindeer.

Natural killer (NK) cells are a vital part of the rapid and non-specific immune defense against invading pathogens and tumor cells. This study evaluated NK cell-like activity by flow cytometry for the first time in three ecologically and culturally important Arctic mammal species: polar bear (Ursus maritimus), muskox (Ovibos moschatus) and reindeer (Rangifer tarandus). NK cell-like activity for all three species was most effective against the mouse lymphoma cell line YAC-1, compared to the human leukemia cell line K562; NK cell response displayed the characteristic increase in cytotoxic activity when the effector:target cell ratio increased. Comparing NK activity between fresh and cryopreserved mouse lymphocytes revealed little to no difference in function, highlighting the applicability of cryopreserving cells in field studies. The evaluation of this important innate immune function in Arctic mammals can contribute to future population health assessments, especially as pollution-induced suppression of immune function may increase infectious disease susceptibility.

ACCURACY OF NONINVASIVE ANESTHETIC MONITORING IN THE ANESTHETIZED GIRAFFE (GIRAFFA CAMELOPARDALIS).

This study evaluated the accuracy of pulse oximetry, capnography, and oscillometric blood pressure during general anesthesia in giraffes (Giraffa camelopardalis). Thirty-two giraffes anesthetized for physiologic experiments were instrumented with a pulse oximeter transmittance probe positioned on the tongue and a capnograph sampling line placed at the oral end of the endotracheal tube. A human size 10 blood pressure cuff was placed around the base of the tail, and an indwelling arterial catheter in the auricular artery continuously measured blood pressure. Giraffes were intermittently ventilated using a Hudson demand valve throughout the procedures. Arterial blood for blood gas analysis was collected at multiple time points. Relationships between oxygen saturation as determined by pulse oximetry and arterial oxygen saturation, between arterial carbon dioxide partial pressure and end-tidal carbon dioxide, and between oscillometric pressure and invasive arterial blood pressure were assessed, and the accuracy of pulse oximetry, capnography, and oscillometric blood pressure monitoring evaluated using Bland-Altman analysis. All three noninvasive methods provided relatively poor estimates of the reference values. Receiver operating characteristic curve fitting was used to determine cut-off values for hypoxia, hypocapnia, hypercapnia, and hypotension for dichotomous decision-making. Applying these cut-off values, there was reasonable sensitivity for detection of hypocapnia, hypercapnia, and hypotension, but not for hypoxemia. Noninvasive anesthetic monitoring should be interpreted with caution in giraffes and, ideally, invasive monitoring should be employed.

Effects of Polar Bear and Killer Whale Derived Contaminant Cocktails on Marine Mammal Immunity.

Most controlled toxicity studies use single chemical exposures that do not represent the real world situation of complex mixtures of known and unknown natural and anthropogenic substances. In the present study, complex contaminant cocktails derived from the blubber of polar bears (PB; Ursus maritimus) and killer whales (KW; Orcinus orca) were used for in vitro concentration-response experiments with PB, cetacean and seal spp. immune cells to evaluate the effect of realistic contaminant mixtures on various immune functions. Cytotoxic effects of the PB cocktail occurred at lower concentrations than the KW cocktail (1 vs 16 µg/mL), likely due to differences in contaminant profiles in the mixtures derived from the adipose of each species. Similarly, significant reduction of lymphocyte proliferation occurred at much lower exposures in the PB cocktail (EC50: 0.94 vs 6.06 µg/mL; P < 0.01), whereas the KW cocktail caused a much faster decline in proliferation (slope: 2.9 vs 1.7; P = 0.04). Only the KW cocktail modulated natural killer (NK) cell activity and neutrophil and monocyte phagocytosis in a concentration- and species-dependent manner. No clear sensitivity differences emerged when comparing cetaceans, seals and PB. Our results showing lower effect levels for complex mixtures relative to single compounds suggest that previous risk assessments underestimate the effects of real world contaminant exposure on immunity. Our results using blubber-derived contaminant cocktails add realism to in vitro exposure experiments and confirm the immunotoxic risk marine mammals face from exposure to complex mixtures of environmental contaminants.

Conjunctival mucinous adenocarcinoma in an ostrich (Struthio camelus).

A 2-year-old male ostrich (Struthio camelus) presented with a rapidly growing soft tissue mass protruding from the ventromedial aspect of the right eye (OD). The initial physical examination revealed a soft tissue mass attached to the medial inferior conjunctiva. The mass was excised with cryosurgery, and the conjunctival tissue margins were treated with cryoablation. Histopathological examination diagnosed granulomatous inflammation associated with scattered acid-fast bacteria. The ostrich recovered uneventfully and appeared healthy until recurrence of a grossly similar mass 2 months later. Gross examination revealed a botryoid mass attached to the inferior palpebral conjunctiva and extending onto the palpebral aspect of the nictitating membrane. Euthanasia was selected, and the histological diagnosis of the second mass was a mixed mucinous adenocarcinoma; however, no acid-fast bacteria were seen. Granulomatous conjunctival lesions have been previously described in the ostrich, while, to the authors' knowledge, neoplastic conjunctival lesions have not. Neoplasia should be considered as a differential diagnosis for a rapidly recurring, granulomatous conjunctival mass in this species.

The thick left ventricular wall of the giraffe heart normalises wall tension, but limits stroke volume and cardiac output.

Giraffes--the tallest extant animals on Earth--are renowned for their high central arterial blood pressure, which is necessary to secure brain perfusion. Arterial pressure may exceed 300 mmHg and has historically been attributed to an exceptionally large heart. Recently, this has been refuted by several studies demonstrating that the mass of giraffe heart is similar to that of other mammals when expressed relative to body mass. It thus remains unexplained how the normal-sized giraffe heart generates such massive arterial pressures. We hypothesized that giraffe hearts have a small intraventricular cavity and a relatively thick ventricular wall, allowing for generation of high arterial pressures at normal left ventricular wall tension. In nine anaesthetized giraffes (495±38 kg), we determined in vivo ventricular dimensions using echocardiography along with intraventricular and aortic pressures to calculate left ventricular wall stress. Cardiac output was also determined by inert gas rebreathing to provide an additional and independent estimate of stroke volume. Echocardiography and inert gas-rebreathing yielded similar cardiac outputs of 16.1±2.5 and 16.4±1.4 l min(-1), respectively. End-diastolic and end-systolic volumes were 521±61 ml and 228±42 ml, respectively, yielding an ejection fraction of 56±4% and a stroke volume of 0.59 ml kg(-1). Left ventricular circumferential wall stress was 7.83±1.76 kPa. We conclude that, relative to body mass, a small left ventricular cavity and a low stroke volume characterizes the giraffe heart. The adaptations result in typical mammalian left ventricular wall tensions, but produce a lowered cardiac output.

COMPARISON OF ETORPHINE-ACEPROMAZINE AND MEDETOMIDINE-KETAMINE ANESTHESIA IN CAPTIVE IMPALA (AEPYCEROS MELAMPUS).

Impala (Aepyceros melampus) are a notoriously difficult species to manage in captivity, and anesthesia is associated with a high risk of complications including mortality. The aim of this study was to compare an opioid-based protocol with an α-2 agonist-based protocol. Nine female impala were studied in a random cross-over design. Subjects received either an etorphine-acepromazine (EA) protocol: 15 µg/kg etorphine and 0.15 mg/kg acepromazine, or a medetomidine-ketamine (MK) protocol: 109 µg/kg medetomidine and 4.4 mg/kg ketamine on day 1. Anaesthesia was repeated 3 days later with the alternative protocol. Subjective assessments of the quality of induction, muscle relaxation, and recovery were made by a blinded observer. Objective monitoring included blood pressure, end-tidal CO2, regional tissue oxygenation, and blood gas analysis. EA provided a significantly quicker (mean EA, 7.17 mins; MK, 17.6 mins) and more-reliable (score range EA, 3-5; MK, 1-5) induction. Respiratory rates were lower for EA with higher end-tidal CO2, but no apnoea was observed. As expected, blood pressures with EA were lower, with higher heart rates; however, arterial oxygenation and tissue oxygenation were equal or higher than with the MK protocol. In conclusion, at these doses, EA provided superior induction and equivalent muscle relaxation and recovery with apparent improved oxygen tissue delivery when compared to MK.

Protection against high intravascular pressure in giraffe legs.

The high blood pressure in giraffe leg arteries renders giraffes vulnerable to edema. We investigated in 11 giraffes whether large and small arteries in the legs and the tight fascia protect leg capillaries. Ultrasound imaging of foreleg arteries in anesthetized giraffes and ex vivo examination revealed abrupt thickening of the arterial wall and a reduction of its internal diameter just below the elbow. At and distal to this narrowing, the artery constricted spontaneously and in response to norepinephrine and intravascular pressure recordings revealed a dynamic, viscous pressure drop along the artery. Histology of the isolated median artery confirmed dense sympathetic innervation at the narrowing. Structure and contractility of small arteries from muscular beds in the leg and neck were compared. The arteries from the legs demonstrated an increased media thickness-to-lumen diameter ratio, increased media volume, and increased numbers of smooth muscle cells per segment length and furthermore, they contracted more strongly than arteries from the neck (500 ± 49 vs. 318 ± 43 mmHg; n = 6 legs and neck, respectively). Finally, the transient increase in interstitial fluid pressure following injection of saline was 5.5 ± 1.7 times larger (n = 8) in the leg than in the neck. We conclude that 1) tissue compliance in the legs is low; 2) large arteries of the legs function as resistance arteries; and 3) structural adaptation of small muscle arteries allows them to develop an extraordinary tension. All three findings can contribute to protection of the capillaries in giraffe legs from a high arterial pressure.

Sensorimotor responsiveness and resolution in the giraffe.

The ability of an animal to detect and respond to changes in the environment is crucial to its survival. However, two elements of sensorimotor control - the time required to respond to a stimulus (responsiveness) and the precision of stimulus detection and response production (resolution) - are inherently limited by a competition for space in peripheral nerves and muscles. These limitations only become more acute as animal size increases. In this paper, we investigated whether the physiology of giraffes has found unique solutions for maintaining sensorimotor performance in order to compensate for their extreme size. To examine responsiveness, we quantified three major sources of delay: nerve conduction delay, muscle electromechanical delay and force generation delay. To examine resolution, we quantified the number and size distribution of nerve fibers in the sciatic nerve. Rather than possessing a particularly unique sensorimotor system, we found that our measurements in giraffes were broadly comparable to size-dependent trends seen across other terrestrial mammals. Consequently, both giraffes and other large animals must contend with greater sensorimotor delays and lower innervation density in comparison to smaller animals. Because of their unconventional leg length, giraffes may experience even longer delays compared with other animals of the same mass when sensing distal stimuli. While there are certainly advantages to being tall, there appear to be challenges as well - our results suggest that giraffes are less able to precisely and accurately sense and respond to stimuli using feedback alone, particularly when moving quickly.

Pheochromocytoma in a white rhinoceros (Ceratotherium simum).

A 46-yr-old male white rhinoceros (Ceratotherium simum) died during anesthesia following agonal excitation. On postmortem, a well-demarcated 2.5-cm tan mass was identified in the right adrenal gland. Histopathology confirmed the presence of a pheochromocytoma, and elevated levels of epinephrine in serum collected shortly prior to the animal's death, as compared with sera from healthy controls, demonstrated the functional nature of the tumor. Although rare, pheochromocytoma should be considered as a differential diagnosis in cases of suspected hypertension and acute death in rhinos.

Identification of acute phase proteins and assays applicable in nondomesticated mammals.

The serum concentration of acute phase proteins (APPs) increases dramatically in response to inflammation and tissue injury. APPs are clinically useful in a range of domesticated mammals; however, knowledge is limited in nondomesticated mammals. The detective ability of two assays for each of three potential APPs--serum amyloid A (SAA), C-reactive protein (CRP), and haptoglobin (Hp)--was evaluated in eight species. For SAA, a turbidimetric immunoassay (TIA) demonstrated significant detective abilities in the Asian elephant (Elaphas maximus), impala (Aepyceros melampus), musk ox (Ovibos moschatus), and chimpanzee (Pan troglodytes), as did an SAA enzyme-linked immunosorbent assay (ELISA) in the impala. For CRP, both TIA and ELISA had significant detective abilities in the chimpanzee. For Hp, a colorimetric assay demonstrated significant detective abilities in impala, musk ox, sitatunga (Tragelaphus spekeii), and chimpanzee, as did the Hp ELISA in the impala, musk ox, and sitatunga. In conclusion, these results suggest that assays for detection of relevant APPs in several nondomesticated animals are available.

Serological response to vaccination against avian influenza in zoo-birds using an inactivated H5N9 vaccine.

Five hundred and forty birds in three zoos were vaccinated twice against avian influenza with a 6-week interval using an inactivated H5N9 vaccine. Serological response was evaluated by hemagglutination inhibition test 4-6 weeks following the second vaccine administration. 84% of the birds seroconverted, and 76% developed a titre > or =32. The geometric mean titre after vaccination was 137. A significant species variation in response was noted; penguins, pelicans, ducks, geese, herons, Guinea fowl, cranes, cockatiels, lovebirds, and barbets showed very poor response to vaccination, while very high titres and seroconversion rates were seen in flamingos, ibis, rheas, Congo peafowl, black-winged stilts, amazon parrots, and kookaburras.

Disseminated Mycobacterium celatum infection in a white-tailed trogon (Trogon viridis).

An adult female white-tailed trogon (Trogon viridis) was presented with abdominal enlargement and hard subcutaneous masses. Necropsy findings included bony masses extending from skeletal structures, disseminated pale foci in the liver, and a pale mass in the kidney. Histological examination revealed multifocal to coalescing granulomatous inflammation in the bone, liver, kidney, lung and spleen. Mycobacterium celatum was isolated from the liver and identified by DNA sequencing. This is the first report of M. celatum infection in an avian species.

Molecular evidence for the infection of zoo chimpanzees by pig Ascaris.

We here describe the transmission of the pig roundworm, Ascaris suum to chimpanzees maintained in the Copenhagen Zoo, Denmark. Using a technique for whole genome fingerprinting, amplified fragment length polymorphism (AFLP) and the technique PCR restricted fragment length polymorphism (PCR-RFLP) of the internal transcribed spacer region (ITS) of the nuclear ribosomal DNA, the worms from the chimpanzees were compared with Ascaris spp obtained from humans and pigs in order to identify the source of the infection. By the use of different distance and clustering based methods on the AFLP data set the worms from the chimpanzees were assigned to the same cluster as that of the worms from pigs. The PCR-RFLP analysis supported the AFLP results. Therefore, the zoo chimpanzees have required Ascaris infections by cross-infection from pigs. Pigs as a potential source of Ascaris infections for both captive and wild chimpanzees and other animals, therefore needs to be considered and appropriate steps taken to prevent such infection.