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Only limited data on concentrations of trace elements in the blood of avian species have been published. This information can play an important role in the conservation of endangered species and their protection from environmental pollutants and can also be clinically relevant in managed individuals. Some elements are essential for the health of the animals in human care, but little is known about expected concentrations for some of these elements. Therefore, 21 elements (silver, arsenic, gold, barium, cadmium, cobalt, chromium, copper, iron, iodine, mercury, magnesium, manganese, molybdenum, nickel, platinum, antimony, selenium, thallium, vanadium, and zinc) were measured by inductively coupled plasma mass spectrometry in lithium-heparinized plasma samples from Humboldt penguins ( Spheniscus humboldti ) from a single zoological collection in Germany ( n = 39). The plasma concentrations of cadmium, nickel, platinum, antimony, and thallium were under the limit of detection in most of the analyzed samples. The results also showed that the females had significantly ( P ≤ 0.05) higher concentrations of silver, barium, cobalt, chromium, iron, mercury, magnesium, vanadium, and zinc in their plasma in comparison with males. A correlation between the element plasma concentration and the age of the individuals could not be found. The differences in plasma concentrations of individual element between males and females could be due to hormonal influences on the metabolism. Further studies in different seasons and other age groups are needed in the future to investigate environmental and host effects in more detail and to understand physiological mechanisms responsible for observed differences.

Leopard sharks ( Triakis semifasciata ) are temperate, Eastern Pacific elasmobranchs popular in public aquariums. Blood analysis is commonly used for assessing animal health, yet reference values have not been established for this species. This study analyzed T. semifasciata population data to characterize blood reference values for a collection of T. semifasciata housed at a public aquarium. Twenty-one captive leopard sharks were sampled. Blood was collected during annual health examinations from sedated animals. After collection, blood samples were anticoagulated with lithium heparin, and hematocrit and plasma biochemistry values were analyzed. The minimum-maximum ranges were hematocrit 11-31%, buffy coat 1-2%, glucose 4.94-9.38 mM/L, sodium 244-272 mM/L, potassium 3.7-5.5 mM/L, chloride 214-246 mM/L, aspartate aminotransferase 5-31 U/L, creatine kinase 36-1,136 U/L, calcium 3.65-3.95 mM/L, phosphorus 1.13-2.23 mM/L, total protein 21-38 g/L, and total CO 2 12-18 mM/L. The values identified will contribute to a better understanding of captive leopard shark physiology and to improved veterinary care for captive leopard sharks. Further research can examine the validity of machines like the Vetscan VS2, which will expand the resources available to care professionals.

Acute phase proteins (APP) and protein electrophoresis (EPH) offer crucial insights into inflammation and overall health in various species. In this study, we validated serum amyloid A (SAA) and C-reactive protein (CRP) reagents for use with serum samples from gibbons (Hylobatidae, n = 50), spanning five species across four gibbon genera: eastern hoolock ( Hoolock leuconedys ), Javan ( Hylobates moloch ), pileated ( Hylobates pileatus ), siamang ( Symphalangus syndactylus ), and white-cheeked ( Nomascus leucogenys ). Preliminary reference intervals ( n = 50) were calculated for SAA (1.8-48.1 mg/L), CRP (0.1-11.1 mg/L), and EPH via capillary zone electrophoresis, in healthy gibbons. Comparing clinically normal ( n = 38) and abnormal ( n = 12) individuals, significant differences were observed in the albumin/globulin ratio ( P = 0.0003), prealbumin ( P = 0.0345), and albumin ( P = 0.0094), with abnormal individuals exhibiting statistically significantly higher γ-globulins ( P = 0.0224), SAA ( P = 0.0001), and CRP ( P = 0.0003). Despite significant chromosomal rearrangements among different gibbon species, we found no statistically significant differences of SAA and CRP levels across species. However, some differences between species were observed in EPH fractions. This study presents the first report of the evaluation of APP and EPH in gibbons, underscoring the potential use of these biomarkers in gibbon health monitoring. Further research with larger sample sizes of both normal and abnormal gibbons is recommended to solidify the clinical utility of these biomarkers in these species.

Protein electrophoresis (PEP) is an important tool in mammals to characterize specific dysproteinemias and detect acute and chronic inflammatory responses. In reptiles, PEP is the gold standard method for globulin fraction determination and albumin measurement. In this study, preliminary reference intervals were established for serum PEP in 22 clinically healthy adult Roti Island snake-necked turtles ( Chelodina mccordi ), a critically endangered species, kept in captivity and sampled over two monsoon seasons. The species has a prominent prealbumin fraction and β-globulins were the dominant globulin fraction. Significant differences between females and males were found in prealbumin ( P < 0.01), albumin ( P = 0.02), α1-globulin ( P = 0.05) and γ-globulin ( P = 0.01). Gravid females had significantly lower total protein ( P < 0.01), prealbumin ( P < 0.01), albumin ( P < 0.01) and albumin:globulin ratio ( P = 0.01). These preliminary reference intervals should aid in clinical investigation in this species as well as further research studies seeking to understand the application of PEP in reptilian species.

This study reports comprehensive clinical pathology data for hematology, serum, and plasma biochemistry reference intervals for 174 apparently healthy common bottlenose dolphins (Tursiops truncatus) and reference values for 27 Indo-Pacific bottlenose dolphins (Tursiops aduncus), 13 beluga whales (Delphinapterus leucas), and 6 Pacific white-sided dolphins (Lagenorhynchus obliquidens) in zoos and aquariums accredited by the Alliance for Marine Mammal Parks and Aquariums and the Association of Zoos & Aquariums. Blood samples were collected as part of a larger study titled "Towards understanding the welfare of cetaceans in zoos and aquariums" (colloquially called the Cetacean Welfare Study). Two blood samples were collected following a standardized protocol, and two veterinarian examinations were conducted approximately six months apart between July to November 2018 and January to April 2019. Least square means, standard deviations, and 95% confidence intervals were calculated for hematology, serum, and plasma biochemical variables. Comparisons by age, gender, and month revealed statistically significant differences (p < 0.01) for several variables. Reference intervals and values were generated for samples tested at two laboratories for up to 56 hematologic, serum, and plasma biochemical variables. To apply these data, ZooPhysioTrak, an iOS mobile software application, was developed to provide a new resource for cetacean management. ZooPhysioTrak provides species-specific reference intervals and values based on user inputs of individual demographic and sample information. These data provide a baseline from which to compare hematological, serum, and plasma biochemical values in cetaceans in zoos and aquariums., Competing Interests: The authors have read the journal’s policy, and the authors of the study have the following competing interests to declare: LJM and LKL are affiliated with the Chicago Zoological Society – Brookfield Zoo (an AZA and AMMPA accredited zoo), and Chicago Zoological Society provides a salary for LJM. Before the study, JDM was previously affiliated with Disney’s Animal Kingdom (an AZA accredited zoo). The Seas® Epcot® Walt Disney World® Resort, Dolphin Island – Resorts World Sentosa, and Loro Parque and Loro Parque Fundación provided funding to the Chicago Zoological Society for this study. This does not alter our adherence to PLOS ONE policies on sharing data and materials. There are no patents, products in development or marketed products associated with this research to declare.

Background: Zoological gardens contain unique configurations of exotic and endemic animals and plants that create a diverse range of developing sites and potential sources of blood meals for local mosquitoes. This may imply unusual interspecific pathogen transmission risks involving zoo vertebrates, like avian malaria to captive penguins. Understanding mosquito ecology and host feeding patterns is necessary to improve mosquito control and disease prevention measures in these environments., Methods: Mosquito sampling took place in Chester Zoo for 3 years (2017, 2018, and 2019) and for 1 year in Flamingo Land (2017) using different trapping methods. Blood-fed mosquitoes were identified and their blood meal was amplified by PCR, sequenced, and blasted for host species identification., Results: In total, 640 blood-fed mosquitoes were collected [Culex pipiens (n = 497), Culiseta annulata (n = 81), Anopheles maculipennis s.l. (n = 7), An. claviger (n = 1), and unidentifiable (n = 55)]. Successful identification of the host species was achieved from 159 blood-fed mosquitoes. Mosquitoes fed on birds (n = 74), non-human mammals (n = 20), and humans (n = 71). There were mixed blood meals from two hosts (n = 6). The proportions of blood-fed mosquitoes varied across sampling seasons and sites within the zoos. The use of resting traps and aspiration of vegetation were more efficient techniques for capturing blood-fed mosquitoes than traps for host-seeking or gravid mosquitoes. By relating the locations of zoo vertebrates to where fed mosquitoes were trapped, the minimum travelling distances were calculated (13.7 to 366.7 m). Temperature, precipitation, relative humidity, proximity to zoo vertebrate exhibits, and vegetation level were found to be significantly associated with the proportion of captured blood-fed mosquitoes by generalized linear modelling., Conclusions: Mosquito feeding behaviour in zoos is mainly influenced by time, location (sampling area), temperature, and host availability, which highlights the value of mosquito monitoring in complex settings to plan control strategies and potentially reduce inherent disease transmission risks for humans and threatened zoo vertebrates.

Haematological and biochemical parameters play important roles in safeguarding animal health and preventing disease, but the blood reference values of many wild animals are still unknown. Recently, few descriptions of the blood parameters of Siberian tigers (Panthera tigris altaica) have been reported because these tigers comprise an endangered species; however, it is extremely difficult to obtain blood samples necessary for these analyses. This study presents 14 haematological and 16 biochemical parameters of 133 Siberian tigers, of which 112 and 21 were from Heilongjiang Siberian Tiger Park (HB) and Hailin Siberian Tiger Park (HD), China, respectively. Our study is the first to determine the following parameters in Siberian tigers: red blood cell volume distribution width, platelet count, mean platelet volume, amylase (AMY), sodium/potassium, globulin and albumin/globulin levels. As the data for total bilirubin and AMY were not statistically significant, no statistical analysis was conducted for these parameters. Few parameters were significantly different according to sex and region (p < 0.05). The concentration of alkaline phosphatase decreased with age, whereas the creatinine (CREA) increased with age. The CREA concentration of tigers raised in HB was much lower than that of tigers raised in HD. The data obtained in this study provide a reference for monitoring the health of wild and captive Siberian tigers and will add important information to the standards for haematological and biochemical parameters of wild felines., (© 2020 The Authors Veterinary Medicine and Science Published by John Wiley & Sons Ltd.)

Background: The cardiovascular system of owl monkeys has been studied due to frequent postmortem findings of heart disease in asymptomatic animals. The silent aspect and the difficulty of early diagnosis intensify the importance of studying the cardiovascular system in this species., Methods: Echocardiogram evaluation was carried out on 60 animals, grouped into suspect or non-suspect of having heart diseases, and evaluated through electrocardiogram, hematology, and biochemical tests., Results: Doppler echocardiography indicated two animals with suspicion of left ventricular hypertrophy and eight with dilated cardiomyopathy. Suspect animals had higher cardiac measurements and reduced shortening fraction. Troponin I was detectable in two animals (0.128 ng/mL and 0.584 ng/mL), and serum albumin concentration was significantly higher in non-suspect animals (P < .05)., Conclusions: The importance of echocardiographic measurements of IVSd, IVSs, LVIDd, LVIDs, LVPWd, LVPWs, LA, EF, and FS in the cardiac evaluation of captive owl monkeys was evidenced., (© 2021 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Tramadol is used as an analgesic in humans and some animal species. When tramadol is administered to most species it undergoes metabolism to its main metabolites M1 or O-desmethyltramadol, and M2 or N-desmethyltramadol, and many other metabolites. This study describes the pharmacokinetic profile of tramadol when a single subcutaneous bolus of 2 mg/kg was initially administered to two koalas. Based on the results of these two koalas, subsequently 4 mg/kg as a single subcutaneous injection, was administered to an additional four koalas. M1 is recognised as an active metabolite and has greater analgesic activity than tramadol, while M2 is considered inactive. A liquid chromatography assay to quantify tramadol, M1 and M2 in koala plasma was developed and validated. Liquid chromatography-mass spectrometry confirmed that M1 had been identified. Additionally, the metabolite didesmethyltramadol was identified in chromatograms of two of the male koalas. When 4 mg/kg tramadol was administered, the median half-life of tramadol and M1 were 2.89 h and 24.69 h, respectively. The M1 plasma concentration remained well above the minimally effective M1 plasma concentration in humans (approximately 36 ng/mL) over 12 hours. The M1 plasma concentration, when tramadol was administered at 2 mg/kg, did not exceed 36 ng/mL at any time-point. When tramadol was administered at 2 mg/kg and 4 mg/kg the area under the curve M1: tramadol ratios were 0.33 and 0.50, respectively. Tramadol and M1 binding to plasma protein were determined using thawed, frozen koala plasma and the mean binding was 20% and 75%, respectively. It is concluded that when tramadol is administered at 4 mg/kg as a subcutaneous injection to the koala, it is predicted to have some analgesic activity., Competing Interests: The authors have declared that no competing interests exist.

Reference intervals of blood analytes are important diagnostic tools that clinicians use as a basis for making clinical decisions and that have been documented for many fish species. Although the effects of dietary supplementation on hematology and blood biochemistry variables in clownfish have been shown, there are no published reference intervals to date for routine hematology and blood biochemistry panels for these popular cultured marine fish (including Amphiprion spp.). The objective of this study was to establish de novo reference intervals for selected hemogram data and, using whole-blood analysis, commonly utilized chemistry analytes in two species of aquacultured clownfish: the Tomato Clownfish Amphiprion frenatus and Saddleback Clownfish A. polymnus. This study is based on blood samples from 25 clinically normal Tomato Clownfish and 38 clinically normal Saddleback Clownfish from two clownfish production facilities in Florida. Guidelines by the American Society for Veterinary Clinical Pathology were followed to generate robust reference intervals from the data collected. We report descriptive statistics and reference intervals for several hematology (n = 12) and biochemistry (n = 8) analytes using a small sample size and a small amount of blood per sample. The blood analyte data from this study are comparable with hematology and biochemistry data that have been reported in other teleost species. These results provide baseline health data for clownfish species that can be used in various clinical settings while also serving as a springboard for further research studies., (© 2020 American Fisheries Society.)

The zoos manage small populations of endangered big cat species like tiger, lion, and leopard for display, research, and conservation breeding. Genetic management of these populations is essential to ensure long term survival and conservation utility. Here we propose a simple and cost effective microsatellite based protocol for the genetic management of captive big cats. We sampled 36 big cat individuals from Seoul Grand Park Zoo (Republic of Korea) and amplified 33 published microsatellite loci. Overall, allelic richness and gene diversity was found highest for leopards, followed by lions and tigers. Twelve of the thirty-three markers showed a high degree of polymorphism across all target species. These microsatellites provide a high degree of discrimination for tiger (1.45 × 10 -8 ), lion (1.54 × 10 -10 ), and leopard (1.88 × 10 -12 ) and thus can be adopted for the genetic characterization of big cats in accredited zoos globally. During captive breeding, zoo authorities rely on pedigree records maintained in studbooks to ensure mating of genetically fit unrelated individuals. Several studies have reported errors in studbook records of big cat species. Microsatellites are simple and cost effective tool for DNA fingerprinting, estimation of genetic diversity, and paternity assessment. Our unified microsatellite panel (12-plex) for big cats is efficient and can easily be adopted by zoo authorities for regular population management.

The aim of this study was to objectively evaluate the biological variation of healthy Asian elephant ( Elephas maximus ) hematology and biochemistry parameters, therefore enabling evidence-based clinical decision-making to improve patient management. Ten clinically healthy elephants had blood samples collected weekly for 5 wk under standardized conditions. The analytical, between- and within-individual variation, index of individuality, and reference change values were calculated using previously reported methods. Large between-individual variation and small within-individual variation for almost all parameters indicated that individual normal values should be used for interpreting blood results from Asian elephants.

Hyperprolactinemia is an endocrine disorder associated with infertility in many species, including elephants. In a recent survey of zoos accredited by the Association of Zoos and Aquariums (AZA), over half of African elephant females (N = 101) were not cycling normally, 30% of which exhibited hyperprolactinemia. We examined whether life experience and temperament predict ovarian cyclicity and circulating prolactin status in individual African elephant females. We hypothesized that, similar to humans, acyclicity and hyperprolactinemia in elephants will be associated with an apprehensive or fearful, anxious temperament, and an increased number of potentially challenging life events (transfers, deaths and births). Ninety-five adult African elephant females housed at 37 AZA institutions were included in this study. Blood samples were collected twice a month for 1 year to determine ovarian cycle (cycling, n = 44; irregular, n = 13; non-cycling, n = 38) and prolactin (normal, n = 44; low; n = 23; high; n = 28) status. Keeper ratings on a 6-point scale were obtained on 32 temperament traits in 85 of these elephants. We determined that giving birth and being exposed to herd mates entering the facility were positively associated with normal ovarian cycle and prolactin profiles. By contrast, age, serum cortisol, and an increased number of herd mates leaving a facility were negatively associated with both. Contrary to our hypothesis, hyperprolactinemia was associated with a popular and caring temperament rating, whereas consistently low prolactin was associated with a fearful, apprehensive temperament. These findings indicate that pituitary-ovarian function may be impacted by life history (cyclicity) and temperament (prolactin), which should be taken into consideration when making management decisions., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Blood samples collected from captive Southern lechwes ( Kobus leche ) were used to develop reference intervals for complete blood count and serum biochemistry values. The study population consisted in 108 healthy individuals, including 62 females and 46 males and 38 neonates (<2 days old), 27 juveniles (2 mo to 2 yr old), and 43 adults (>2 yr old) from the Réserve Africaine de Sigean, Sigean, France, between 2014 and 2016. No significant effects of sex and season of sampling were found. No significant differences were found for all hematology and biochemistry parameters between juveniles and adults, and their results were then combined. Significant differences were found between animals >2 mo old and neonates for most tested parameters; in particular, neonates had lower hemoglobin, eosinophil count, alanine transaminase, and total protein values, but higher alkaline phosphatase values. The reference intervals obtained were similar to values previously reported for other Bovidae species.

Pronghorn ( Antilocapra americana ) are native to western North America and are found in 24 Association of Zoos and Aquariums (AZA)-accredited institutions. Acute-phase proteins (APP) are a broad class of proteins that are stimulated in response to inflammation and have been shown to be a sensitive measure of inflammation in equids and ruminants. In this study, blood samples from clinically normal free-ranging and captive populations of pronghorn were analyzed using assays for protein electrophoresis (EPH) and APP, including serum amyloid A (SAA) and haptoglobin (HP), to develop preliminary ranges to gauge potential differences between these populations. Additional samples were taken from clinically abnormal captive pronghorn with facial abscesses. By EPH measurements, albumin: globulin ratio mean and SE were significantly different ( P <0.05) with 1.02 (0.08) for captive populations and 1.91 (0.05) for free-ranging populations. Total protein mean and SE were significantly different ( P <0.05) for captive and free-ranging populations, respectively 5.6 (0.3) g/dl and 6.9 (0.1) g/dl. Mean and SD of SAA for captive pronghorn were 1.4 (3.2) mg/L, and were significantly different from the free-ranging population, which was below the limits of detection for ( P <0.05). There was no difference in HP levels between these groups. In a case study of a pronghorn with facial abscesses, elevated levels of HP, but not SAA, suggested that HP maybe useful in certain disease states. Future studies should explore the use of these biomarkers as tools to monitor general health, prognosis, and subclinical disease.

This study assessed the in vitro temporal changes that occur in blood pH and lactate concentrations for an elasmobranch species and a chelonian species, as well as blood gases (partial pressures of carbon dioxide [pCO 2 ] and oxygen [pO 2 ]) for a chelonian species, with a portable clinical point-of-care analyzer. Blood samples were collected from 10 cownose rays ( Rhinoptera bonasus ) and 10 red-eared sliders ( Pseudemys scripta elegans ), stored on ice, and serially analyzed at six time points up to 90 min postcollection. Results indicate that analysis should be conducted as soon as possible after blood collection for these species, with immediate analysis being preferred. However, if analysis must be delayed, syringes may be capped, placed on ice, and analyzed at a later time. Analysis within 90 min provided clinically acceptable results for pH and lactate in both species and for pCO 2 in red-eared sliders, whereas substantial artifactual increases of pO 2 were seen in red-eared sliders.

The reference intervals of health parameters are valuable tools for veterinarians and conservationists to monitor the health status and viability of endangered species. Natural variation in the health of the long-lived Asian elephant ( Elephas maximus ) is poorly understood, particularly in relation to differences between males and females. Longitudinal health data were collected from clinical examination, hematology, and serum chemistry analyses over 3 yr from 227 healthy individually marked Asian elephants varying in age and sex. The study population was semicaptive and used in Myanmar's timber industry, but maintained natural feeding and breeding behavior. Body condition score (BCS) and blood pressure were investigated in clinical examinations. Hematological parameters included hematocrit, hemoglobin, total white blood cell count, and differential blood cell counts. Serum chemistry parameters included blood urea nitrogen, creatinine, total protein, albumin, globulins, aspartate aminotransferase, alkaline phosphatase, triglycerides, creatine kinase, glucose, calcium, potassium, sodium, and chloride. To the knowledge of the authors, this is the first description of BCS in an elephant population outside of zoos, and of blood pressure in this species using a novel adaptation of the Intelli Wrap Cuff pressure monitor. Several differences between the sexes were observed, with females generally having higher BCS and triglycerides, and males displaying higher alkaline phosphatase and glucose levels. This study provides important clinical tools that can be used to assess the health status and improve management in this endangered species.

Background: Culicoides biting midges are biological vectors of arboviruses including bluetongue virus (BTV), Schmallenberg virus (SBV) and African horse sickness virus (AHSV). Zoos are home to a wide range of 'at risk' exotic and native species of animals. These animals have a high value both in monetary terms, conservation significance and breeding potential. To understand the risk these viruses pose to zoo animals, it is necessary to characterise the Culicoides fauna at zoos and determine which potential vector species are feeding on which hosts., Methods: Light-suction traps were used at two UK zoos: the Zoological Society of London (ZSL) London Zoo (LZ) and ZSL Whipsnade Zoo (WZ). Traps were run one night each week from June 2014 to June 2015. Culicoides were morphologically identified to the species level and any blood-fed Culicoides were processed for blood-meal analysis. DNA from blood meals was extracted and amplified using previously published primers. Sequencing was then carried out to determine the host species., Results: A total of 11,648 Culicoides were trapped and identified (n = 5880 from ZSL WZ; n = 5768 from ZSL LZ), constituting 25 different species. The six putative vectors of BTV, SBV and AHSV in northern Europe were found at both zoos and made up the majority of the total catch (n = 10,701). A total of 31 host sequences were obtained from blood-fed Culicoides. Culicoides obsoletus/C. scoticus, Culicoides dewulfi, Culicoides parroti and Culicoides punctatus were found to be biting a wide range of mammals including Bactrian camels, Indian rhinoceros, Asian elephants and humans, with Culicoides obsoletus/C. scoticus also biting Darwin's rhea. The bird-biting species, Culicoides achrayi, was found to be feeding on blackbirds, blue tits, magpies and carrion crows., Conclusions: To our knowledge, this is the first study to directly confirm blood-feeding of Culicoides on exotic zoo animals in the UK and shows that they are able to utilise a wide range of exotic as well as native host species. Due to the susceptibility of some zoo animals to Culicoides-borne arboviruses, this study demonstrates that in the event of an outbreak of one of these viruses in the UK, preventative and mitigating measures would need to be taken.

In brown kiwi (Apteryx mantelli), the male is the primary incubator, a trait that is relatively rare among birds. The maintenance of avian incubation behavior is controlled by the protein hormone prolactin (PRL). Although steroid hormone concentrations in both wild and captive kiwi have previously been reported, this study is the first to report levels of PRL in captive and wild male and female kiwi through the prebreeding and breeding seasons, and to directly compare testosterone (T) concentrations between captive and wild males during the breeding and incubation periods. Female PRL concentrations increased at the time of oviposition, whereas male PRL concentrations rose gradually between the prebreeding and incubation periods. Although males are considered the main incubator, an increase in PRL levels could help females maintain behaviors such as nest guarding, or to take over incubation the event of mate loss. A gradual increase in PRL allows the male to be ready for incubation during the long breeding season. Interestingly, T concentrations in captive males did not decrease during incubation and was significantly higher than in wild males. Continual elevated T could have an impact on sperm production through negative feedback, thereby contributing to the low egg fertility seen in captive kiwi. Therefore, determining the underlying reason for the differences in hormone levels could be significant, if not vital, for improving the success of captive kiwi breeding programs., (© 2019 Wiley Periodicals, Inc.)

This experiment was conducted to study the effect of gradual replacement of dietary buffalo meat on the bone (BMB) with chicken carcass (CC) on nutrient utilization, serum cortisol, and total serum antioxidant profile of zoo-housed Indian leopard. Twelve adult leopards were randomly distributed into a replicated Latin square design comprising three treatments, three periods, four animals, and three sequences. Leopards in group T 1 were fed normal zoo diet of BMB. On the basis of dry matter, 10% and 20% of BMB was replaced with CC in groups T 2 and T 3 , respectively. Each experimental period comprised 21 days. During each period, a digestion trial of 4-day collection period was conducted after an adaptation period of 17 days. On Day 21 of each experimental period, blood was collected from all the animals by puncturing the ventral coccygial vein. Intake and apparent digestibility of major nutrients were similar among the groups. Replacement of 20% BMB with addition of CC increased (p < 0.001) the calculated supply of I, niacin, and vitamin A. Carotenoid intake increased (p < 0.01) with increased level of CC in the diet. Serum concentration of cortisol decreased (p < 0.01) whereas serum concentration of total carotenoids increased (p < 0.001) with increased level of CC in the diet. Serum concentration of antioxidant enzymes increased (p < 0.001) with increased level of CC in the diet. It was concluded that replacement of 20% of BMB with CC increased antioxidant profile. This may reduce oxidative stress in zoo-housed Indian leopards without any adverse effect on nutrient utilization., (© 2019 Wiley Periodicals, Inc.)

The black-fronted piping-guan ( Aburria jacutinga ) is and an endangered cracid species and little is known about its health. Hematology allows researchers to assess the health and physiological status of a population, but, to our knowledge, reference intervals are not available in the literature for A. jacutinga . Thus, the present study aimed to determine the hematological reference intervals of 31 captive black-fronted pipingguan adults in the state of Paraná, Brazil. Blood samples were collected from brachial vein and stored in heparinized tubes to be processed. The red blood cell (RBC) and white blood cell (WBC) counts were performed in a hemocytometer, and hemoglobin (HGB) concentration was obtained by the spectrophotometric cyanmethemoglobin method; packed cell volume (PCV) was determined by microhematocrit technique, and the mean corpuscular volume (MCV) and mean corpuscular hemoglobin concentration (MCHC) were calculated according to a previously established formula. The heterophil : lymphocyte ratio was calculated by dividing the number of heterophils by the number of lymphocytes. Determination of the 95% reference intervals and 90% confidence intervals were performed in accordance with American Society for Veterinary Clinical Pathology guidelines using Reference Value Advisor (version 2.0). The results obtained were as follows: RBC, 1.4-2.8 × 10 6 /µl; PCV, 34.3-51.2%; HGB, 8.0-12.6 g/dl; MCV, 157.3-261.4 fl; MCHC, 22.0-26.7 g/dl; WBC, 2.3-33.1 × 10 3 /µl; heterophils, 12.1-16.3%, 0.5-14.4 × 10 3 /µl; lymphocytes, 19.8-69.0%, 1.4-16.4 × 10 3 /µl; monocytes, 0.7-17.3%, 0.1-2.8×10 3 /µl; eosinophils, 0.4-12.8%, 0.0-3.9 × 10 3 /µl; basophils, 0.8-12.0%, 0.0-1.9 × 10 3 /µl; and the heterophil : lymphocyte ratio, 0.2-3.0. The present study fills gaps in the literature on hematological values for A. jacutinga , which can be used to assess the health of the species populations. In addition, it contributed to establishing the health status of captive black-fronted piping-guans, indicating the possible welfare of the birds., (© 2019 by American Association of Zoo Veterinarians.)

Ill and anorectic captive sharks present a unique challenge for husbandry and veterinary staff. Providing adequate fluid and nutritional support to sharks while minimizing handling remains difficult. This study aimed to evaluate the ability of a nutrient enema to alter blood analyte concentrations. Thirty-six healthy, fasted white-spotted bamboo sharks ( Chiloscyllium plagiosum ) were enrolled in the study with 18 sharks receiving a nutrient enema and 18 sharks receiving a non-nutrient saline enema. The metabolic state of sharks was evaluated via measurement of blood glucose, blood urea nitrogen, and β-hydroxybutyrate as well as other serum biochemistry parameters. Changes in sodium, chloride, calcium, β-hydroxybutyrate, glucose, total protein, and triglyceride concentrations were seen across time in both groups. Blood glucose absolute concentrations and changes over time differed between the nutrient and nonnutrient groups. This pilot study indicates that it is possible to influence the glucose metabolism of healthy sharks via nutrient enema. Further study is needed to better understand potential therapeutics for ill and anorectic sharks., (© 2019 by American Association of Zoo Veterinarians.)

Trace nutrients have been shown to play important health roles in both domestic and wild animals, but there has been little investigation into their values and effects in giant pandas ( Ailuropoda melanoleuca ). The goal of this study was to obtain serum vitamin and mineral values in a group of healthy, captive giant pandas ( n = 4). Twenty-seven samples were obtained from the group from 2008 to 2012. Serum analytes measured included retinol, α-tocopherol, hydroxyvitamin D, cobalt (Co), copper (Cu), iron (Fe), manganese (Mn), molybdenum (Mo), selenium (Se), and zinc (Zn). The diet was evaluated for protein, fat, fiber, Ca, P, magnesium (Mg), potassium (K), sodium (Na), Fe, Zn, Cu, Mn, and Mo. Comparison with other giant panda nutrient values were made. Further research with increased sample sizes and different populations are warranted., (© 2019 by American Association of Zoo Veterinarians.)

Pangolins are one of the most-trafficked endangered species and are rare in zoologic collections. Blood gas and select biochemical analysis was performed on nine African white-bellied tree pangolins ( Phataginus tricuspis). Paired blood samples were collected from the ventral tail vasculature prior to (T1) and after 10 min (T2) of sevoflurane anesthesia, and selected blood gas and biochemical parameters were measured using a portable blood gas analyzer. Median (min-max) values for corrected venous blood were pH = 7.372 (7.207-7.529) at T1 and 7.278 (7.124-7.418) at T2. Corrected values were PCO 2 = 51.7 (29.6-69.0) mmHg at T1 and 58.431 (38.1-90.8) mmHg at T2. Clinicians should be aware that this species may hypoventilate while anesthetized with sevoflurane and develop a respiratory acidosis. Concomitant administration of respiratory depressants could compound this effect in a species where oro-tracheal intubation has not been described.

Black rhinoceros ( Diceros bicornis spp.) are critically endangered species, with less than 65 individual animals housed in captivity within Association of Zoos and Aquariums-accredited zoos within the United States, and an estimated 5,500 individual animals of all subspecies surviving in the wild. Previously published reference values for circulating vitamin D 3 (25OHD 3 ; 55.7 ± 34.2 ng/ml) were based upon samples from free-ranging black rhinoceros in Africa. Recent research in human medicine has highlighted the importance of subclinical vitamin D deficiency, with links to increased risks for developing various health conditions. Serum samples collected opportunistically from two captive Eastern black rhinoceros ( Diceros bicornis michaeli) housed with seasonal access outdoors in a North American zoo were tested for 25-hydroxy-vitamin D (25OHD) levels over a 3-yr period. A commercially prepared pelleted diet containing vitamin D 3 was fed to both rhinos. This study correlates environmental ultraviolet (UV) index, dietary supplementation, and seasonal serum 25OHD levels to compare with known 25OHD 3 levels in free-ranging African black rhinoceros. Results in these two individuals suggest that D. bicornis spp. are dependent upon sunlight or UVB for measurable circulating 25OHD, and that current vitamin D 3 supplementation levels may have little effect for Diceros spp. in human care housed in northern latitudes.

During this retrospective study, 18 plasma blood chemistry and 17 complete blood count (CBC) samples were analyzed from clinically healthy spotted eagle rays ( Aetobatus narinari) at Georgia Aquarium in order to generate hematological ranges for complete blood count (CBC) and biochemical profiles. Summary statistics were generated according to the American Society for Veterinary Clinical Pathology guidelines for the determination of reference intervals in veterinary species. 4 The mean packed cell volume (PCV) was 28.09% with a range of 23-35%. Mean total solids were 5.72 g/dl with a range of 5-7.0 g/dl. Lymphocytes were the dominant leukocyte observed on differential (67.35%), followed by fine eosinophilic granulocytes (FEGs) (15.41%), coarse eosinophilic granulocytes (CEGs) (10.24%), monocytes (1.88%), and basophils (1.24%). Chemistry samples were analyzed at two diagnostic laboratories, Michigan State University (MSU) and University of Miami (UMiami), and the results were compared. Both labs have the capacity to run blood chemistries on zoo and aquatic species, but utilize different methods to obtain chemistry analyte values. UMiami uses a thin-film dry-slide technology, whereas MSU uses an ion-selective electrode (ISE) and Beckman Coulter AU 640 analyzer. There is poor agreement between the analyzers used by the two laboratories for both alkaline phosphatase and BUN, because of proportional error. Establishing hematological ranges in spotted eagle rays and in elasmobranchs in general may enhance the understanding of the species and their health. This information may aid clinicians in deciding when and how to treat elasmobranchs.

Toxoplasma gondii is protozoan parasite with ability of causing disease in wide-spectrum of animals; many species of animals in captivity died of clinical toxoplasmosis. The monitoring of T. gondii antibodies in zoo animals can be an important indicator of T. gondii circulation in zoo. The aim of this study was to examine sera of animals from eight Czech zoos by latex agglutination test with statistical evaluation and detect T. gondii DNA in stray cats and rodents captured in the zoos. T. gondii antibodies were detected in 33% of 1043 zoo animals without statistical difference between birds (27%, n = 74) and mammals (33%, n = 969). In birds, the chance to be infected with T. gondii was higher in Accipitriformes (71%) compared to Pelecaniformes (6%) (p < 0.0001). In mammals, the chance to be infected with T. gondii was higher in Carnivora (63%) compared to Cetarodactyla (30%), Perissodactyla (26%), Primates (28%) and Rodentia (13%) (p < 0.0001) and higher in Felidae (70%) compared to Bovidae (28%) and Equidae (28%) (p < 0.0001). Mammals with carnivore/scavenger way of feeding were in a higher risk of T. gondii infection compared to herbivores and omnivores (p < 0.0001). T. gondii DNA was detected in tissue of one stray cat while in none of 77 rodents caught in zoo. This study is the first report on toxoplasmosis in zoos from the Czech Republic including seroepidemiology and molecular detection., (Copyright © 2018. Published by Elsevier B.V.)

To determine the effect of capture and restraint on hematologic, plasma biochemical, and venous blood gas values in Dalmatian pelicans ( Pelecanus crispus), blood samples were collected and temperature and heart rate were recorded in 13 pelicans (5 males and 8 females) immediately after capture from a large field exhibit. Repeat sampling was performed after transport, a mean of 74 minutes after initial evaluation. When compared with values at initial sampling to the time of capture, significant increases were documented in the heterophil : lymphocyte ratio (H : L), carbon dioxide (CO 2 ), bicarbonate (HCO 3 ), total carbon dioxide (Tco 2 ), base excess (BE), and partial pressure of carbon dioxide (Pco 2 ) values after 74 minutes. Significant decreases were documented in total protein, lactate, and phosphorus concentrations, percentage of oxygen saturation (So 2 ), partial pressure of oxygen (Po 2 ), total white blood cell count (WBC), absolute lymphocytes, absolute eosinophils, and absolute monocytes after 74 minutes. Those changes suggest capture and restraint can alter hematologic, plasma biochemical, and acid-base status in Dalmatian pelicans, which may influence clinical decision making and case management.

Increased glucocorticoids cause a characteristic stress leukogram in mammalian taxa. It is assumed that avians exhibit a similar response, but to date, there have been no controlled studies to correlate serial endogenous corticosterone levels to hematologic values. An established flock of 18 Hispaniolan Amazon parrots ( Amazona ventralis) was used as a model in a crossover study. The treatment group was subjected to the stress of transport, restraint, and common clinical procedures with serial blood samples collected at 20-min intervals for hematology and corticosterone levels; the control group was sampled at the same intervals. Longitudinal data analysis was performed with linear mixed modeling. For all hematologic analytes, the baseline value had a significant positive effect on subsequent values (all P < 0.001). The white blood cell, heterophil, and eosinophil counts and heterophil to lymphocyte ratio increased over time in the treatment group, whereas it remained stable in the control group ( P = 0.016, P < 0.001, P < 0.001, P = 0.02, respectively, for the time*treatment effect). Lymphocyte absolute counts decreased over time, although not significantly; the decrease was significant for the relative lymphocyte count in the treatment group. Monocytes and basophils were not significantly altered. The treatment group had a higher mean corticosterone level overall than the control group by approximately 60% ( P = 0.008). The mean corticosterone level also increased over time in both groups by three- to fourfold ( P < 0.001) by 20 min then plateaued. These results demonstrate that some significant hematologic changes may arise with routine handling and transportation of birds and should be accounted for in hematologic interpretation of cell counts.

Health evaluations were conducted on individuals ( n = 62, age < 1 to 11 yr) from a captive red panda ( Ailurus fulgens styani) population at the Chengdu Research Base for Giant Panda Breeding in Sichuan Province, People's Republic of China. Individuals were anesthetized using combinations of ketamine, dexmedetomidine, diazepam, isoflurane, and tiletamine/zolazepam. Whereas physical exams revealed no abnormalities for 40% of red pandas, 24% had dental abnormalities and 18% had orthopedic abnormalities. As the red panda is the only member of the family Ailuridae, complete blood count and serum biochemistry samples were collected and used to develop reference intervals for this species. When hematologic and serum chemistry values of male and female red pandas were compared within this A. f. styani population, males had higher mean cell hemoglobin, mean cell hemoglobin concentration, and blood urea nitrogen concentrations, whereas females had higher alkaline phosphatase, cholesterol, and iron. Adult red pandas (≥2 yr) had higher lymphocyte percentage, total protein, globulin, and chloride levels, whereas juveniles (<2 yr) had higher white blood cell concentrations, neutrophil percentage and absolute neutrophil values, alkaline phosphatase, creatine kinase, cholesterol, potassium, total and ionized calcium, and phosphorus levels. Reference values and expected age and sex differences will assist with health management of the captive A. f. styani population.

Iron overload disorder (IOD) can lead to organ dysfunction and may exacerbate other diseases in the critically endangered black rhinoceros ( Diceros bicornis). It is important to develop methods for monitoring the progression of iron storage (hemosiderosis), diagnosing the disease, and evaluating treatments in this species. Traditionally, an equine enzyme immunoassay (EIA) was used to measure rhinoceros ferritin, a serum protein correlated to iron stores. The goal of this study was to validate a rhinoceros-specific assay and investigate factors potentially associated with ferritin concentrations in black rhinoceros. A ferritin EIA developed for Sumatran rhinoceros was validated for black rhinoceros via Western blot analysis of liver ferritin and confirmed parallelism of serum samples to the EIA standard curve and used to analyze serum samples ( n = 943) collected from 36 black rhinoceros (<1-33 yr) at 14 U.S. institutions. Mean (±SEM) serum ferritin concentration was 6,738 ± 518 ng/ml (range: 85-168,451 ng/ml). Concentrations differed among individuals with eastern black rhinoceros (7,444 ± 1,130 ng/ml) having a higher mean ferritin than southern black rhinoceros (6,317 ± 505 ng/ml; P < 0.05) and higher mean values in wild-born (11,110 ± 1,111 ng/ml) than captive-born individuals (3,487 ± 293 ng/ml; P < 0.05). Ferritin concentrations did not differ between young rhinoceros (<5 yr old; 2,163 ± 254 ng/ml) and adults (7,623 ± 610 ng/ml) and were not correlated with age ( r = 0.143) or time in captivity ( r = 0.146, wild born; r = 0.104, all animals). Ferritin concentration was not impacted by sex (female: 2,086 ± 190 ng/ml; male: 8,684 ± 717 ng/ml), date, month, or season of collection ( P > 0.05). Data indicate ferritin concentrations are variable and not necessarily associated with IOD; ferritin is not recommended for diagnosing or monitoring IOD in black rhinoceros.

We monitored annual fluctuations of gonadal steroid levels in three sexually mature captive finless porpoises (Neophocaena asiaeorientalis; two males and one female) from two different facilities over 56-91 months. Two animals (one male and one female) were held in an indoor tank with a sunroof (facility A) and the other male was held in an indoor tank without a sunroof (facility B). Water temperatures in both facilities reflected seasonal changes during the study period with a minor difference in the fluctuation pattern. Testosterone levels of the male in facility A were higher from spring to summer every year and exhibited a 12-month cycle. The female showed estrus cycles in 1-month intervals from summer to winter, excluding 2 anestrus years. In contrast, the period of higher testosterone levels of the male in facility B gradually initiated earlier over the years under a constant photoperiod (11.5L:12.5D) and exhibited a 9-month cycle during the first 52 months. After changing the light conditions to a natural photoperiod, its testosterone levels were high from early spring to summer for 3 consecutive years and exhibited a 12-month cycle. Our results showed that under a constant artificial photoperiod, the male in facility B failed to recognize the seasonal changes of a natural external environment, resulting in a 9-month, free-running hormone cycle.

Background: Hematologic and serum biochemical reference values obtained from captive or free-ranging wildlife populations may not be comparable as there can be significant variations due to preanalytic and analytic differences, including methods of capture and restraint, overall management in captivity including diet and composition of animal groups, and analytic methods being used. Hematology and serum biochemistry have never been studied in captive or free-ranging populations of Sechuran foxes (Lycalopex sechurae)., Objectives: The purposes of the study were to determine hematologic and serum biochemical RI in Sechuran foxes and to explore differences in these variables related to sex and overall life circumstances., Methods: Blood samples were obtained from 15 free-ranging and 15 captive Sechuran foxes. Hematology variables were assessed by blood smear examination and automated analyzer methodology. Serum biochemical analysis was performed by automated analyzer methodology. Descriptive statistics were calculated for each variable. Data obtained from free-ranging and captive groups were statistically compared and RIs were calculated., Results: Captive Sechuran foxes had significantly (P < .05) higher MCH, MCHC, and eosinophil counts and significantly lower band neutrophil counts than free-ranging foxes. Free-ranging Sechuran foxes had significantly (P < .05) higher serum lipase and globulins and significantly lower albumin, total bilirubin, and indirect bilirubin than captive foxes., Conclusions: These findings suggest that there are hematologic and serum biochemical differences between captive and free-ranging Sechuran fox populations. Hence, such differences should be considered when using these variables to assess the health status of this species., (© 2018 American Society for Veterinary Clinical Pathology.)

Fifteen giant pandas (Ailuropoda melanoleuca) from the Chinese Conservation and Research Center for the Giant Panda (CCRCGP) in Bifengxia, Sichuan, China were analyzed for blood lead concentrations (Pb-B) during the 2017 breeding season. Thirteen of the 15 bears showed Pb-B below the method detection limit (MDL) of 3.3 µg/dL. The two remaining bears, although above the MDL, contained very low concentrations of lead of 3.9 and 4.5 µg/dL. All 15 giant pandas in this analysis had Pb-B concentrations that were within normal background concentrations for mammals in uncontaminated environments. For a threatened species, whose native country is plagued by reports of extremely high air pollution, our findings suggest that giant pandas at the CCRCGP are not absorbing lead at concentrations that would adversely affect their health.

This experiment was conducted to study the effect of feeding graded levels of dietary crude protein (CP) on serum biochemical profile of Lady Amherst's pheasants (LAP). Eighteen male LAP were randomly distributed into three groups of six each in an experiment based on completely randomized design. The CP content of the diets was 13.4%, 16.5%, and 19.1%, in groups I, II, and III, respectively. Serum concentrations of uric acid was lowest (P < 0.05) in group I. Relationship between serum concentration of uric acid and nitrogen intake was linear (R(2)  = 0.39, P < 0.01). Concentrations of other serum metabolites and enzymes were similar among the groups. Serum concentration of triiodothyronine (T3 ) was highest (P < 0.05) in group I, followed by groups II and III. Total antioxidant capacity (TAC) of serum was lower (P < 0.007), whereas serum concentration of malondialdehyde (MDA) was higher (P < 0.001) in group I as compared to groups II and III. Regression of serum concentration of TAC (R(2)  = 0.74, P < 0.01) and MDA (R(2)  = 0.39, P < 0.05) was polynomial. Heterophil to lymphocyte ratio was higher (P < 0.007) in group I as compared to groups II and III. Relationship between H/L ratio and nitrogen intake was polynomial (R(2)  = 0.69, P < 0.05). Cell mediated immune response measured as foot web index was similar among the groups. Based upon the results, it was concluded that a diet containing 16.5% crude protein would be optimum for improving antioxidant defense and the ability of Lady Amherst's pheasant to combat stress. Zoo Biol. 35:346-354, 2016. © 2016 Wiley Periodicals, Inc., (© 2016 Wiley Periodicals, Inc.)

Early identification of illness and/or presence of environmental and/or social stressors in free-ranging and domestic cetaceans is a priority for marine mammal health care professionals. Incorporation of leukocyte gene transcript analysis into the diagnostic tool kit has the potential to augment classical diagnostics based upon ease of sample storage and shipment, inducible nature and well-defined roles of transcription and associated downstream actions. Development of biomarkers that could serve to identify "insults" and potentially differentiate disease etiology would be of great diagnostic value. To this end, a modest number of peripheral blood leukocyte gene transcripts were selected for application to a domestic killer whale population with a focus on broad representation of inducible immunologically relevant genes. Normalized leukocyte transcript values, longitudinally acquired from 232 blood samples derived from 26 clinically healthy whales, were not visibly influenced temporally nor by sex or the specific Park in which they resided. Stability in leukocyte transcript number during periods of health enhances their potential use in diagnostics through identification of outliers. Transcript levels of two cytokine genes, IL-4 and IL-17, were highly variable within the group as compared to the other transcripts. IL-4 transcripts were typically absent. Analysis of transcript levels on the other genes of interest, on an individual animal basis, identified more outliers than were visible when analyzed in the context of the entire population. The majority of outliers (9 samples) were low, though elevated transcripts were identified for IL-17 from 2 animals and one each for Cox-2 and IL-10. The low number of outliers was not unexpected as sample selection was intentionally directed towards animals that were clinically healthy at the time of collection. Outliers may reflect animals experiencing subclinical disease that is transient and self-limiting. The immunologic knowledge derived from longitudinal immunologic studies in killer whales, as was the target of the present study, has the potential to improve diagnostics and health related decision making for this and other domestic and free-ranging cetacean species., (Copyright © 2016. Published by Elsevier B.V.)

Monthly blood samples, daily mating observations from Galapagos tortoises (Chelonoidis nigra), and local rainfall and temperature were collected at the Honolulu Zoo as part of a fertility evaluation. Testosterone concentrations were measured for males (n = 6), two of which were seen copulating and were considered sexually active. Estrone sulfate and progesterone concentrations were measured for female tortoises (n = 9), two of which nested and only one had laid eggs. Testosterone profiles were similar for both sexually active and sexually inactive males, both of which were positively correlated with temperature but not rainfall. Peak testosterone concentrations (12.0 ± 1.4 ng/ml sexually active animals vs. 14.4 ± 2.4 ng/ml sexually inactive animals) occurred at the end of the nesting season, from April to July. Estrone sulfate concentrations were similar for nesting (n = 2) and non-nesting (n = 7) female tortoises, rising from non-detectable concentrations (September), and increasing to peak concentrations during the nesting season. Progesterone concentrations remained low and spiked (9.44 ng/ml) only for the female that nested and laid eggs. Testosterone was negatively correlated with mating behavior, and the male tortoises were likely capable of spermatogenesis even though only two of them engaged in mating behavior. The female tortoises were not senescent, as the estrone sulfate concentrations likely reflected waves of ovarian follicular activity. Endocrine parameters were not in synchrony with rainfall, and a disconnect between the timing of reproductive events and the environmental milieu may help to explain the poor fertility of these tortoises. Zoo Biol. 35:237-245, 2016. © 2016 Wiley Periodicals, Inc., (© 2016 Wiley Periodicals, Inc.)

Published haematologic and serum biochemistry reference intervals are very scarce for captive cheetahs and even more for free-ranging cheetahs. The current study was performed to establish reference intervals for selected serum biochemistry analytes in cheetahs. Baseline serum biochemistry analytes were analysed from 66 healthy Namibian cheetahs. Samples were collected from 30 captive cheetahs at the AfriCat Foundation and 36 free-ranging cheetahs from central Namibia. The effects of captivity-status, age, sex and haemolysis score on the tested serum analytes were investigated. The biochemistry analytes that were measured were sodium, potassium, magnesium, chloride, urea and creatinine. The 90% confidence interval of the reference limits was obtained using the non-parametric bootstrap method. Reference intervals were preferentially determined by the non-parametric method and were as follows: sodium (128 mmol/L - 166 mmol/L), potassium (3.9 mmol/L - 5.2 mmol/L), magnesium (0.8 mmol/L - 1.2 mmol/L), chloride (97 mmol/L - 130 mmol/L), urea (8.2 mmol/L - 25.1 mmol/L) and creatinine (88 µmol/L - 288 µmol/L). Reference intervals from the current study were compared with International Species Information System values for cheetahs and found to be narrower. Moreover, age, sex and haemolysis score had no significant effect on the serum analytes in this study. Separate reference intervals for captive and free-ranging cheetahs were also determined. Captive cheetahs had higher urea values, most likely due to dietary factors. This study is the first to establish reference intervals for serum biochemistry analytes in cheetahs according to international guidelines. These results can be used for future health and disease assessments in both captive and free-ranging cheetahs.

Honey badgers (Mellivora capensis) prey upon and survive bites from venomous snakes (Family: Elapidae), but the molecular basis of their venom resistance is unknown. The muscular nicotinic cholinergic receptor (nAChR), targeted by snake α-neurotoxins, has evolved in some venom-resistant mammals to no longer bind these toxins. Through phylogenetic analysis of mammalian nAChR sequences, we show that honey badgers, hedgehogs, and pigs have independently acquired functionally equivalent amino acid replacements in the toxin-binding site of this receptor. These convergent amino acid changes impede toxin binding by introducing a positively charged amino acid in place of an uncharged aromatic residue. In venom-resistant mongooses, different replacements at these same sites are glycosylated, which is thought to disrupt binding through steric effects. Thus, it appears that resistance to snake venom α-neurotoxin has evolved at least four times among mammals through two distinct biochemical mechanisms operating at the same sites on the same receptor., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Arenaviruses are one of the largest families of human hemorrhagic fever viruses and are known to infect both mammals and snakes. Arenaviruses package a large (L) and small (S) genome segment in their virions. For segmented RNA viruses like these, novel genotypes can be generated through mutation, recombination, and reassortment. Although it is believed that an ancient recombination event led to the emergence of a new lineage of mammalian arenaviruses, neither recombination nor reassortment has been definitively documented in natural arenavirus infections. Here, we used metagenomic sequencing to survey the viral diversity present in captive arenavirus-infected snakes. From 48 infected animals, we determined the complete or near complete sequence of 210 genome segments that grouped into 23 L and 11 S genotypes. The majority of snakes were multiply infected, with up to 4 distinct S and 11 distinct L segment genotypes in individual animals. This S/L imbalance was typical: in all cases intrahost L segment genotypes outnumbered S genotypes, and a particular S segment genotype dominated in individual animals and at a population level. We corroborated sequencing results by qRT-PCR and virus isolation, and isolates replicated as ensembles in culture. Numerous instances of recombination and reassortment were detected, including recombinant segments with unusual organizations featuring 2 intergenic regions and superfluous content, which were capable of stable replication and transmission despite their atypical structures. Overall, this represents intrahost diversity of an extent and form that goes well beyond what has been observed for arenaviruses or for viruses in general. This diversity can be plausibly attributed to the captive intermingling of sub-clinically infected wild-caught snakes. Thus, beyond providing a unique opportunity to study arenavirus evolution and adaptation, these findings allow the investigation of unintended anthropogenic impacts on viral ecology, diversity, and disease potential.

Hematologic and plasma biochemistry parameters of barn owls (Tyto alba) were studied in collaboration by the Exotic Division of the Faculty of Veterinary Science of the Szent Istvan University and the Eötvös Loránd University, both in Budapest, Hungary. Blood samples were taken from a total of 42 adult barn owls kept in zoos and bird repatriation stations. The following quantitative and qualitative hematologic values were determined: packed cell volume, 46.2 +/- 4%; hemoglobin concentration, 107 +/- 15 g/L; red blood cell count, 3.2 +/- 0.4 x 10(12)/L; white blood cell count, 13.7 +/- 2.7 x 10(9)/L; heterophils, 56.5 +/- 11.5% (7.8 +/- 2 x 10(9)/L); lymphocytes, 40.3 +/- 10.9% (5.5 +/- 1.9 x 10(9)/L); monocytes, 1.8 +/- 2.1% (0.3 +/- 0.3 x 10(9)/ L); eosinophils, 1 +/- 1% (0.1 +/- 0.1 x 10(9)/L); and basophils, 0.6 +/- 0.5% (0.1 +/- 0.1 x 10(9)/L). The following plasma biochemistry values also were determined: aspartate aminotransferase, 272 +/- 43 U/L; L-gamma-glutamyltransferase, 9.5 +/- 4.7 U/L; lipase, 31.7 +/- 11.1 U/L; creatine kinase, 2228 +/- 578 U/L; lactate dehydrogenase, 1702 +/- 475 U/L; alkaline phosphatase, 358 +/- 197 U/L; amylase, 563 +/- 114 U/L; glutamate dehydrogenase, 7.5 +/- 2.5 U/L; total protein, 30.6 +/- 5.3 g/L; uric acid, 428 +/- 102 micromol/L; and bile acids, 43 +/- 18 micromol/L. These results provide reliable reference values for the clinical interpretation of hematologic and plasma biochemistry results for the species.

The ex situ Indian rhino population experienced a decrease in genetic diversity indicating that the breeding program could possibly benefit from novel reproductive management strategies to ensure population sustainability. We sought to determine how management tools used for reproductive management, specifically translocation and operant conditioning, impact physiological and behavioral measures of welfare in Indian rhinos. First, an adrenocorticotropic hormone challenge performed in an adult male resulted in a 38-fold increase in urinary and a 3.5-fold increase in fecal glucocorticoid metabolites (FGM). Mean and peak FGM differed among three females, but all demonstrated elevated (P < 0.0001) concentrations for variable durations after translocation that lasted up to 9 weeks. Lastly, behavioral and adrenal responses of two females to operant conditioning to stand during transrectal ultrasound exams were monitored and rhinos differed in their mean and peak FGM concentrations. However, FGM were not different before versus during training or on pasture versus in the barn. One female exhibited more stereotypic behavior during training in the barn than on pasture (P < 0.05); although, stereotypies (1.73% of time) were relatively uncommon overall. In summary, individual variation exists in FGM both at baseline levels and in response to a stressor. In addition, while a transient rise in glucocorticoid activity post-translocation indicated that Indian rhinos have a physiological response to changes in their environment, minor alterations in daily routines using operant conditioning only resulted in minimal changes in behaviors and FGM., (© 2014 Wiley Periodicals Inc.)

The horned guan (Oreophasis derbianus) is an endangered species with small and seriously fragmented wild populations. Breeding efforts during the last decade have maintained a slowly increasing captive population with the potential to play an important role in the recovery of the species. Clinical hematology and biochemistries are powerful tools to diagnose and monitor diseases in captive birds. Therefore, establishing hematologic and plasma biochemistry reference values will improve the medical management of this species. This study determines the reference values for 9 hematologic and 15 plasma biochemical variables for 27 male and 12 female, apparently healthy, captive horned guans from three institutions. Differences related to age, sex, and husbandry were identified but were usually small and clinically insignificant. These results should improve veterinary care of captive individuals of this species and provide comparative data for other species of cracids.

Background: Usutu virus (USUV), a mosquito-borne flavivirus of the Japanese encephalitis virus antigenic group, caused bird die-offs in Austria, Hungary and Switzerland between 2001 and 2009. While the zoological gardens of Vienna and Zurich recorded USUV-associated mortality in different species of birds during this period, incidences in Budapest were limited to areas outside the zoo, and in the greater Basel area avian mortality due to USUV infection was not observed at all. The objectives of this investigation were to gain insight into USUV infection dynamics in captive birds in zoos with varying degrees of virus exposure and to study differences in susceptibility to USUV of different species of birds., Results: 372 bird sera were collected between October 2006 and August 2007. The samples were tested in parallel by hemagglutination inhibition (HI) and 90% plaque reduction neutralization tests (PRNT-90). 8.75%, 5.3% and 6.59% of birds in the zoos of Vienna, Zurich and Basel, respectively, showed USUV-specific antibodies by PRNT-90. No antibodies to USUV were detected in birds of the Budapest zoo. The order Strigiformes (owls) exhibited the highest USUV-seroprevalence, compared to other orders of birds., Conclusions: USUV seems not to pose an imminent threat to zoo bird populations in central Europe at the moment. Depending on a variety of especially environmental factors, however, this may change at any time in the (near) future, as experienced with West Nile virus (WNV). It is therefore strongly suggested to continue with combined WNV and USUV surveillance activities in affected areas.

Golden conures or ararajubas (Guaruba guarouba) are endangered parrots endemic to the Brazilian Amazon forest. Body mass, blood cell counts, and total plasma protein were determined for 70 clinically healthy golden conures captive at zoologic parks and private breeder facilities in Brazil. Hematologic results (mean +/- SD) were: Erythrocytes 3.6 +/- 0.5 x 10(6) cells/mm3, hemoglobin 12.8 +/- 1.4 g/dl, packed cell volume 46 +/- 3.8%, mean corpuscular volume 132 +/- 20 fl, mean corpuscular hemoglobin (MCH) 36 +/- 5.7 pg, mean corpuscular hemoglobin concentration (MCHC) 28 +/- 3.5%, thrombocytes 26.3 +/- 9.3 x 10(3) cells/mm3, leukocytes 11.9 +/- 4.5 x 10(3) cells/mm3, heterophils 6284 +/- 2715 cells/mm3, lymphocytes 5473 +/- 2408 cells/ mm3, monocytes 113 +/- 162 cells/mm3, eosinophils 10 +/- 42 cells/mm3, basophils 27 +/- 64 cells/mm3. Body mass was 254 +/- 24.9 g and total plasma protein (TPP) was 3.54 +/- 0.58 g/dl. No statistical differences were observed between genders within age groups. Differences between juveniles (J) and adults (A) were identified for TPP (J < A), MCH (J > A), and MCHC (J > A). These results provide reliable reference values for the clinical interpretation of hematologic results for the species. Hematology may be an important tool for population health investigations on free-ranging golden conure populations and will also be essential to survey the health of release candidates in future reintroduction programs.

Bottlenose dolphins can have iron overload (that is, hemochromatosis), and managed populations of dolphins may be more susceptible to this disease than are wild dolphins. Serum iron, total iron-binding capacity (TIBC), transferrin saturation, and ferritin were measured in 181 samples from 141 dolphins in 2 managed collections and 2 free-ranging populations. Although no iron indices increased with age among free-ranging dolphins, ferritin increased with age in managed collections. Dolphins from managed collections had higher iron, ferritin, and transferrin saturation values than did free-ranging dolphins. Dolphins with high serum iron (exceeding 300 μg/dL) were more likely to have elevated ferritin but not ceruloplasmin or haptoglobin, demonstrating that high serum levels of iron are due to a true increase in total body iron. A time-series study of 4 dolphins with hemochromatosis that were treated with phlebotomy demonstrated significant decreases in serum ferritin, iron, and TIBC between pre- and posttreatment samples; transferrin saturation initially fell but returned to prephlebotomy levels by 6 mo after treatment. Compared with those in managed collections, wild dolphins were 15 times more likely to have low serum iron (100 μg/dL or less), and this measure was associated with lower haptoglobin. In conclusion, bottlenose dolphins in managed collections are more likely to have greater iron stores than are free-ranging dolphins. Determining why this situation occurs among some dolphin populations and not others may improve the treatment of hemochromatosis in dolphins and provide clues to causes of nonhereditary hemochromatosis in humans.

Objective: Use haematology, biochemistry and protein electrophoresis analyses to establish reference values for, and describe the health status of, wild and captive colonies of critically endangered warru (black-footed rock-wallaby: Petrogale lateralis MacDonnell Ranges race)., Methods: Blood samples were taken from warru in three wild colonies (Alalka, Kalka, New Well) in the A nangu Pitjantjatjara Yankunytjatjara Lands in north-west South Australia (SA) and from captive animals at Monarto Zoo, SA. General haematology, serum biochemistry and protein electrophoresis analyses were conducted and results used to establish reference ranges. For the parameters that are indicative of a population's health, comparisons among the study sites were completed using analysis of variance., Results: General haematology results suggested that warru were not experiencing chronic anaemia and the protein electrophoresis values indicated that colonies were not suffering from population-wide disease. However, the lower superoxide dismutase, retinol, total carotenoids and ascorbic acid values for New Well warru suggested those animals had a lower plane of nutrition than those at Kalka and Alalka. Higher urea concentrations in New Well and Alalka warru could be a reflection of the absence of reliable free water at these sites., Conclusion: The results have implications for the management of in situ colonies, including potentially using supplementary feeding to improve nutrition, and suggested that these animals were not suffering from disease. The study presents the first blood reference values for P. lateralis and, potentially, a methodology for other threatened species recovery programs to follow to establish the health of their populations., (© 2012 The Authors. Australian Veterinary Journal © 2012 Australian Veterinary Association.)

We obtained whole-blood hematologic and serum biochemical values from 38 captive-bred cotton-top tamarins (Saguinus oedipus). Data were analyzed to determine the effect of sex on blood parameters. Significant differences between either the means or medians of male and female tamarins were found for creatinine, hematocrit, hemoglobin, RBC count, and PCV. These results establish baseline hematologic and serum biochemical values and provide a useful resource not previously available in the peer-reviewed literature for the clinical care of cotton-top tamarins, a critically endangered New World primate, in a captive setting.

Hypocalcemia is a well known cause of dystocia in animals, including elephants in captivity. In order to study calcium metabolism in elephants, it is of utmost importance to use properly validated assays, as these might be prone to specific matrix effects in elephant blood. The aim of the current study was to conduct preliminary work for validation of various parameters involved in calcium metabolism in both blood and urine of captive elephants. Basal values of these parameters were compared between Asian elephants (Elephas maximus) and African elephants (Loxodonta africana). Preliminary testing of total calcium, inorganic phosphorus, and creatinine appeared valid for use in plasma and creatinine in urine in both species. Furthermore, measurements of bone alkaline phosphatase and N-terminal telopeptide of type I collagen appeared valid for use in Asian elephants. Mean heparinized plasma ionized calcium concentration and pH were not significantly affected by 3 cycles of freezing and thawing. Storage at 4 °C, room temperature, and 37 °C for 6, 12, and 24 hr did not alter the heparinized plasma ionized calcium concentration in Asian elephants. The following linear regression equation using pH (range: 6.858-7.887) and ionized calcium concentration in heparinized plasma was utilized: iCa(7.4) (mmol/l) = -2.1075 + 0.3130·pH(actual) + 0.8296·iCa(actual) (mmol/l). Mean basal values for pH and plasma in Asian elephant whole blood were 7.40 ± 0.048 and 7.49 ± 0.077, respectively. The urinary specific gravity and creatinine concentrations in both Asian and African elephants were significantly correlated and both were significantly lower in Asian elephants., (© 2011 The Author(s))