Comparison of three hematocrit measurement methods in the southern white rhinoceros (Ceratotherium simum simum)

BACKGROUND: Hematocrit (HCT) determination is an integral part of health and disease assessments in captive and wild white rhinoceroses. Several affordable automated hematology analyzers have been developed for in-clinic and field use and have the advantage of being able to measure a large number of additional measurands. However, the accuracy of these analyzers for rhinoceros HCT measurements has not yet been investigated. OBJECTIVES: We aimed to compare the HCT results generated by the EPOC portable analyzer system and the Abaxis VetScan HM5 with the gold standard of a manual packed cell volume (PCV) measured using the microhematocrit method. METHODS: Hematocrits were measured with the EPOC and the Abaxis VetScan HM5 (bovine setting) and compared with the PCVs of 69 white rhinoceros whole blood samples. Results were compared using Bland–Altman difference plots and PassingBablok regression analysis. A total allowable analytical error of 10% was set as the performance goal. RESULTS: A significant positive bias, with a mean of 7.7% for the EPOC and 17.9% for the Abaxis, was found compared with the manual PCV method. CONCLUSIONS: The allowable error goal of 10% was not exceeded with the EPOC analyzer. Although not analytically equivalent to the gold standard, the EPOC results could therefore be used as approximations in critical situations where manual measurements cannot be performed. The Abaxis exceeded this allowable error and overestimated HCTs in rhinoceroses. Therefore, method-specific reference intervals should be used.International Rhino Foundation; Veterinary Wildlife Services, Kruger National Park; Zebra Foundation for Veterinary Zoological Education; South African National Parks; University of Pretoriahttp://www.wileyonlinelibrary.com/journal/vcpdm2022Centre for Veterinary Wildlife StudiesCompanion Animal Clinical StudiesParaclinical Science

The effects of feeding and transport length on the welfare of white rhinoceroses (Ceratotherium simum simum) during long-distance translocations : a preliminary study

Translocation is a valuable conservation tool, but poses significant risks for the transported rhinoceroses. Interventions reducing these risks are required to ensure positive welfare during transportation. The aim of this study was to evaluate the effect of journey duration and feeding during the transport of white rhinoceroses (Ceratotherium simum simum). A total of 32 animals were transported by road during two events, five days apart. Fifteen rhinoceroses in the first transport event (37.0 ± 2.4 hr duration) were not fed, while 17 rhinoceroses in the second event (32.2 ± 1.5 hr duration) were offered lucerne. Blood samples were collected at capture and after transport for the evaluation of changes in serum clinical chemistry analytes. The Wilcoxon rank-sum test was used to compare differences between the groups. In all rhinoceroses, transport resulted in changes in serum electrolyte, metabolite and enzyme concentrations, indicating a loss in total body water, nutritional shifts, stress and fatigue. Fed rhinoceroses, transported over a shorter time, displayed greater changes in osmolality (p < 0.006), serum sodium and chloride concentrations (p = 0.005 and = 0.001, respectively) indicating a greater degree of total body water loss than non-fed rhinoceroses. Feeding and a shorter transport duration reduced, but did not prevent, nutritional challenges. A greater increase in the muscle enzymes CK and AST (p = 0.027 and = 0.001, respectively), indicated greater fatigue in non-fed rhinoceroses transported over a longer time. Further work to distinguish the effects of feeding and journey duration is required to better understand the role feeding may play in mitigating welfare challenges during rhinoceros translocation.The International Rhino Foundation rhino research grant, a donation from the South African Veterinary Association Wildlife Group Research Fund, as well as departmental funding.http://www.jsava.co.zaam2023Centre for Veterinary Wildlife Studie

Cardiopulmonary parameters and arterial blood gases during etorphine-medetomidine-midazolam immobilization in free-ranging black rhinoceroses (Diceros bicornis) undergoing electro-ejaculation : a preliminary study

Conservation management interventions for the critically endangered black rhinoceros (Diceros bicornis) require immobilization, which offer opportunities for semen collection and cryopreservation to establish genetic reservoirs. In free-ranging rhinoceroses, a combination of the potent opioid etorphine and the tranquilizer azaperone is routinely used for chemical immobilization but is associated with muscle rigidity and severe cardiopulmonary changes. Additionally, azaperone inhibits semen emission. Seven free-ranging, male, sexually mature black rhinoceroses were immobilized with an alternative protocol consisting of 4.5mg etorphine, 5mg medetomidine, 50mg midazolam and 2,500 IU hyaluronidase delivered remotely by darting from a helicopter. During the immobilization, electro-ejaculation was performed with a portable electro-ejaculator, and a species-specific rectal probe. Animals were observed for muscle tremors. Longitudinal changes in respiratory rate, heart rate and peripheral oxyhemoglobin saturation, measured at 5min intervals, were assessed using a general mixed model. Non-invasive oscillometric blood pressure and arterial blood gas variables were measured at first handling and before reversal and compared using the Wilcoxon rank sum test. All animals were successfully immobilized, showed no muscle tremors, presented with normal heart rates and lactate concentration (<5 mmol/L), recovered uneventfully, but experienced acidemia, hypoxemia and hypercapnia. Induction time and total time in recumbency were 4.2 ± 0.41 and 38.4 ± 6.9min, respectively. Electro-stimulation commenced after 11.7 ± 3.98min and completed after 24.3 ± 6.65min. Semen-rich fractions were successfully collected from six animals. Our observations indicate that etorphinemedetomidine- midazolam provides a promising immobilization protocol for free-ranging black rhinoceroses, that allows for successful electro-ejaculation.Rhino Force SA NPChttps://www.frontiersin.org/journals/veterinary-science#am2022Production Animal Studie

Arterial blood gases and cardiorespiratory parameters in etorphine-medetomidine-midazolam immobilized free-ranging and game-farmed Southern white Rhinoceroses (Ceratotherium simum simum) undergoing electro-ejaculation

With the rapid loss of individuals in the wild, semen cryopreservation has gained importance to safeguard the genetic diversity of white rhinoceroses (Ceratotherium simum). For semen collection via electro-ejaculation, immobilization of free-ranging individuals requires the potent opioid etorphine, which is routinely combined with azaperone, but causes hypoxemia, hypercarbia, acidemia, muscle rigidity, tachycardia, and systemic hypertension. In this study, the suitability of two alternative immobilization protocols including etorphine, medetomidine, and midazolam at different doses (high vs. low etorphine) was evaluated in adult white rhinoceros bulls in two different management systems (free-ranging vs. game-farmed) and undergoing electro-ejaculation. Fourteen free-ranging (Group 1) and 28 game-farmed rhinoceroses (Group 2) were immobilized with ≈2.5 µg/kg etorphine (high dose), ≈2.5 µg/kg medetomidine, ≈25 µg/kg midazolam and 1,500–1,700 IU hyaluronidase and received ≈2.5 µg/kg of butorphanol intravenously at first handling. Twenty game-farmed animals (Group 3) received ≈1 µg/kg etorphine (low dose), ≈5 µg/kg medetomidine, ≈25 µg/kg midazolam and 1,700 IU hyaluronidase. Respiratory rate, heart rate and peripheral hemoglobin oxygen saturation (SpO2) were measured at 5-min intervals; non-invasive oscillometric blood pressures and arterial blood gases at first handling and before reversal of the immobilization; serum clinical chemistry analytes and hematocrit at first handling. Generalized mixed models (fixed factors: group, time, recumbency; random factor: individual rhinoceros) were applied to compare longitudinal changes between free-ranging and game-farmed rhinoceroses immobilized with the higher etorphine dose (Groups 1 and 2), and between the two protocols tested in the game-farmed rhinoceroses (Groups 2 and 3). All animals were successfully immobilized, presented with normal lactate concentrations (<5 mmol/L), experienced no muscle tremors and recovered uneventfully. Hypoxemia and hypertension persisted throughout the immobilization in all groups. Acidemia and hypercarbia were absent in Group 1, but present in the game-farmed animals. The lower etorphine dose in Group 3 resulted in significantly longer induction times, however, tachycardia was not observed. SpO2 was higher for sternal vs. lateral recumbency. Semen-rich fractions were recovered following electro-stimulation in 46 out of the 62 animals. Our findings suggest that etorphine-medetomidine-midazolam provides effective immobilization with fewer side effects compared to previous reports in white rhinoceroses and is suitable for successful electro-ejaculation.Rhino Force SA NPChttps://www.frontiersin.org/journals/veterinary-sciencedm2022Mammal Research InstituteProduction Animal Studie

A comparison of hematological, immunological, and stress responses to capture and transport in wild White Rhinoceros Bulls (Ceratotherium simum simum) supplemented with azaperone or midazolam

Capture and transport are essential procedures for the management and conservation of southern white rhinoceroses (Ceratotherium simum simum), but are associated with stress-induced morbidity and mortality. To improve conservation efforts, it is crucial to understand the pathophysiology of rhinoceros stress responses and investigate drug combinations that could reduce these responses. In this study we measured rhinoceros stress responses to capture and transport by quantifying hematological and immunological changes together with adrenal hormone concentrations. We investigated whether the potent anxiolytic drug midazolam was able to mitigate these responses compared to azaperone, which is more commonly used during rhinoceros transport. Twenty three wild white rhinoceros bulls were transported for 6 h (280 km) within the Kruger National Park for reasons unrelated to this study. Rhinoceroses were immobilized with either etorphine-azaperone (group A, n = 11) or etorphine-midazolam (group M, n = 12) intramuscularly by darting from a helicopter. Azaperone (group A) or midazolam (group M) were re-administered intramuscularly every 2 h during transport. Serial blood samples were collected at capture (TC), the start of transport (T0) and after 6 h of transport (T6). Changes in hematological and immunological variables over time and between groups were compared using general mixed models. Increases in plasma epinephrine and serum cortisol concentrations indicated that rhinoceroses mounted a stress response to capture and transport. Packed cell volume decreased from TC to T6 indicating that stress hemoconcentration occurred at TC. Neutrophils progressively increased and lymphocytes and eosinophils progressively decreased from T0 to T6, resulting in an increase in neutrophil to lymphocyte ratio; a characteristic leukocyte response to circulating glucocorticoids. A reduction in serum iron concentrations may suggest the mounting of an acute phase response. Rhinoceroses experienced a decrease in unsaturated fatty acids and an increase in lipid peroxidation products at capture and toward the end of transport indicating oxidative stress. Midazolam, at the dose used in this study, was not able to mitigate adrenal responses to stress and appeared to directly influence leukocyte responses.The International Rhino Foundation, the Cayman Supporting Women in Research Grant, the Wildlife Group of the South African Veterinary Association, the South African National Research Foundation and the research fund of the Department of Paraclinical Sciences of the Faculty of Veterinary Science, University of Pretoria.https://www.frontiersin.org/journals/veterinary-science#pm2021Centre for Veterinary Wildlife StudiesCompanion Animal Clinical StudiesParaclinical Science

Isoflurane maintenance of neuroanaesthesia in two dogs with hydrocephalus and syringohydromyelia undergoing ventriculoperitoneal shunt surgery

A two-year-old spayed female great dane crossbreed dog and a three-year-old spayed female Yorkshire terrier were presented for surgical correction of hydrocephalus and syringohydromyelia. Obstruction of cerebrospinal fluid flow and associated increase in cerebrospinal fluid volume commonly result in an elevated intracranial pressure. During anaesthesia, maintaining an optimal balance between intracranial pressure and cerebral perfusion is crucial to avoid neuronal damage. Volatile anaesthetics are the most commonly used drugs for maintaining anaesthesia in veterinary practice but have the potential to cause an increase in intracranial pressure through vasodilation. This case series describes inhalational maintenance of neuroanaesthesia for ventriculoperitoneal shunt surgery with isoflurane. Interventions applied to manipulate the anaesthetic drug’s effects on intracranial pressure to maintain optimal cerebral perfusion are described.https://bvajournals.onlinelibrary.wiley.com/journal/20526121hj2021Companion Animal Clinical Studie

Midazolam alters acid-base status less than azaperone during the capture and transport of Southern White Rhinoceroses (Ceratotherium simum simum)

Acidemia represents a major life-threatening factor during rhinoceros capture. The acid-base status during rhinoceros transport is unknown. The purpose of this study was to describe changes in acid-base status during rhinoceros capture and transport and compare these changes between rhinoceroses sedated with azaperone or midazolam. Twenty-three wild white rhinoceros bulls were road-transported 280 km for reasons unrelated to this study. Rhinoceroses were captured with etorphine-azaperone (Group A) or etorphine-midazolam (Group M). During transport, azaperone (Group A) or midazolam (Group M) was re-administered every 2 h and venous blood collected. Changes in blood pH and associated variables were compared over time and between groups using a general linear mixed model. Rhinoceroses of both groups experienced a respiratory and metabolic acidosis during capture (pH 7.109 ± 0.099 and 7.196 ± 0.111 for Group A and Group M, respectively) that was quickly compensated for by the start of transport (pH 7.441 ± 0.035 and 7.430 ± 0.057) and remained stable throughout the journey. Rhinoceroses from Group M showed a smaller decrease in pH and associated variables at capture than rhinoceroses from Group A (p = 0.012). The use of midazolam instead of azaperone could therefore improve the success of rhinoceros capture and thus, contribute to the outcome of important conservation translocations.International Rhino Foundation (IRF); Wildlife Group of the South African Veterinary Association; South African National Research Foundation; Department of Paraclinical Sciences of the Faculty of Veterinary Science, University of Pretoria.http://www.mdpi.com/journal/animalspm2020Centre for Wildlife ManagementCompanion Animal Clinical StudiesParaclinical Science

Seasonal Variations in Heart Rate Variability as an Indicator of Stress in Free-Ranging Pregnant Przewalski's Horses (E. ferus przewalskii) within the Hortobágy National Park in Hungary

Background: Ecosystems with seasonal fluctuations in climate and food availability present physiological challenges to resident mammals and may cause “stress.” The two predominant physiological responses to stressors are (1) the activation of the hypothalamic-pituitary-adrenal axis and (2) the modulation of the autonomic nervous system. To date, the primary indicator for “stress” in wildlife- and zoo animal research are glucocorticoid levels. By measuring the autonomic regulation of cardiac activity, particularly the vagal tone, heart rate variability (HRV) is presently emerging as a suitable indicator of “stress” in farm- and domestic animal research.Objective: The aim of this study was to use HRV, a novel method in wildlife research, to assess seasonal patterns of “stress” in a group of free-ranging Przewalski's horses (Equus ferus przewalskii).Methods: Six pregnant Przewalski's horses from one harem within the Hortobágy National Park in Hungary were subjected to the study. We used a dedicated telemetry system consisting of a subcutaneously implanted transmitter and a receiver and storage unit in a collar to record HRV, heart rate (HR), subcutaneous body temperature, and activity throughout a one-year study period—climate data was also collected. We defined “stress” as a decrease in parasympathetic nervous system tone and calculated RMSSD (root mean square of successive differences) as a measure of HRV. Linear mixed effects models with random intercept per individual were used for statistical analysis.Results: HRV and HR varied considerably throughout the year. Similar to temperate ruminants and hibernating mammals, Przewalski's horses experienced lower HR and HRV during winter, when resources are limited indicating decreased metabolic rates coupled with “stress.” In spring, we observed a drop of HRV along with a peak in HR indicating an increase of allostatic load that is most likely associated with increased energy demands during pregnancy and/or seasonal routines such as the adjustment of the gastrointestinal system to better quality diet.Conclusion: Measuring telemetric HRV is a proven method to study undisturbed reactions of wild animals to their changing environment over the long term. Przewalski's horses experience a loss of complexity in cardiovascular dynamics over the winter and particularly during spring, indicating seasonal “stress.

Etorphine-Ketamine Constant Rate Infusion for Maintenance of Anaesthesia in a Compromised White Rhinoceros (Ceratotherium simum)

A subadult white rhinoceros bull presented for oesophageal endoscopic evaluation and foreign body removal under general anaesthesia. The animal had a history of nasal and oral regurgitation of water and ingesta with weight-loss for 6 days prior to the procedure and had been diagnosed with oesophageal obstruction caused by a bailing wire. Anaesthesia was induced with intramuscular etorphine and azaperone delivered remotely by dart, followed by an intravenous bolus of ketamine. The trachea was intubated, and anaesthesia was maintained with an etorphine-ketamine constant rate infusion (CRI). The rhinoceros did not respond predictably to induction of anaesthesia and developed life-threatening systemic hypotension throughout the 90-minute procedure. A mega-vertebrate demand ventilator was successfully used to provide intermittent positive pressure ventilation when the rhinoceros developed apnoea. This case report describes the maintenance of anaesthesia of a white rhinoceros using an etorphine-ketamine CRI and the causes and management of hypotension and respiratory impairment observed in this patient

Challenges to animal welfare associated with capture and long road transport in boma-adapted black (Diceros bicornis) and semi-captive white (Ceratotherium simum) rhinoceroses

Capture and transport are part of translocation and expose animals to a variety of stressors that can lead to morbidity and mortality. We aimed to establish a better understanding of the physiologic responses to capture and transport in black (Diceros bicornis) and white (Ceratotherium simum) rhinoceroses in Southern Africa. Fourteen adult black rhinoceroses were transported 600 km by vehicle and 32 white rhinoceroses (24 adults and 8 juveniles) were transported 1,300 km by vehicle. The black rhinoceroses had been wild-caught and boma-adapted over 6 wk prior to the translocation and were only sedated to allow for loading into the transport crates. The white rhinoceroses originated from a game farm and were chemically immobilized from a helicopter and then loaded. Paired blood samples were collected from animals at loading (capture) and after transport and evaluated for changes in clinical chemistry analytes, acute phase reactants, and oxidative stress biomarkers. The Wilcoxon rank sum test was used to compare changes in measured analytes from capture and after transport. All rhinoceroses survived capture and transport. Rhinoceroses experienced total body water loss, mobilization of energy reserves, and muscular damage. Alterations in acute phase reactants suggested that animals mounted a stress response. Oxidative stress was observed in black rhinoceroses. We identified the following challenges to animal welfare during transport: hydration status, energy balance, skeletal muscle fatigue, and stress-induced immunomodulation. Measures to mitigate these challenges, such as administration of fluids, need to be included in the planning of future translocations.The Department of Paraclinical Sciences of the University of Pretoria, National Research Foundation, and the Wildlife Group of the South African Veterinary Association.http://www.jwildlifedis.orgam2021Centre for Veterinary Wildlife StudiesCompanion Animal Clinical StudiesParaclinical Science