Health reference intervals and values for common bottlenose dolphins (Tursiops truncatus), Indo-Pacific bottlenose dolphins (Tursiops aduncus), Pacific white-sided dolphins (Lagenorhynchus obliquidens), and beluga whales (Delphinapterus leucas).

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.

Blood-feeding ecology of mosquitoes in two zoological gardens in the United Kingdom.

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.

Pharmacokinetic profile of injectable tramadol in the koala (Phascolarctos cinereus) and prediction of its analgesic efficacy.

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.

Biochemistry, Doppler echocardiography, and electrocardiography evaluation in captive owl monkeys (Aotus sp).

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.

Microsatellite characterization and development of unified STR panel for big cats in captivity: a case study from a Seoul Grand Park Zoo, Republic of Korea.

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.

Haematological and Biochemical Parameters of Captive Siberian Tigers (Panthera tigris altaica) from the Heilongjiang Province, China.

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.

Hematology and Biochemistry Panel Reference Intervals for Captive Saddleback Clownfish Amphiprion polymnus and Tomato Clownfish A. frenatus.

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.

Ovarian cyclicity and prolactin status of African elephants (Loxodonta africana) in North American zoos may be influenced by life experience and individual temperament.

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.

ESTABLISHMENT OF ACUTE-PHASE PROTEIN AND SERUM PROTEIN ELECTROPHORESIS PRELIMINARY REFERENCE VALUES FOR PRONGHORN (ANTILOCAPRA AMERICANA).

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.

HEMATOLOGY AND SERUM BIOCHEMISTRY VALUES OF CAPTIVE SOUTHERN LECHWES (KOBUS LECHE).

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.

SEX DIFFERENCES IN THE REFERENCE INTERVALS OF HEALTH PARAMETERS IN SEMICAPTIVE ASIAN ELEPHANTS (ELEPHAS MAXIMUS) FROM MYANMAR.

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.

TEMPORAL STABILITY OF IN VITRO VENOUS BLOOD GAS, pH, AND LACTATE VALUES OF COWNOSE RAYS (RHINOPTERA BONASUS) AND RED-EARED SLIDER TURTLES (PSEUDEMYS SCRIPTA ELEGANS).

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 [pCO2] and oxygen [pO2]) 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 pCO2 in red-eared sliders, whereas substantial artifactual increases of pO2 were seen in red-eared sliders.

Culicoides species composition and molecular identification of host blood meals at two zoos in the UK.

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.

BIOLOGICAL VARIATION OF HEMATOLOGY AND BIOCHEMISTRY PARAMETERS FOR THE ASIAN ELEPHANT (ELEPHAS MAXIMUS), AND APPLICABILITY OF POPULATION-DERIVED REFERENCE INTERVALS.

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.

EFFECT OF A NUTRIENT ENEMA ON SERUM NUTRIENT CONCENTRATIONS IN WHITE-SPOTTED BAMBOO SHARKS (CHILOSCYLLIUM PLAGIOSUM).

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 ß-hydroxybutyrate as well as other serum biochemistry parameters. Changes in sodium, chloride, calcium, ß-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.

SERUM TRACE NUTRIENT VALUES IN FOUR CAPTIVE GIANT PANDAS (AILUROPODA MELANOLEUCA).

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.

HEMATOLOGICAL REFERENCE INTERVALS OF ENDANGERED CAPTIVE BLACK-FRONTED PIPING-GUANS (ABURRIA JACUTINGA) IN THE STATE OF PARANÁ, BRAZIL.

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 × 106/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 × 103/µl; heterophils, 12.1-16.3%, 0.5-14.4 × 103/µl; lymphocytes, 19.8-69.0%, 1.4-16.4 × 103/µl; monocytes, 0.7-17.3%, 0.1-2.8×103/µl; eosinophils, 0.4-12.8%, 0.0-3.9 × 103/µl; basophils, 0.8-12.0%, 0.0-1.9 × 103/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.

Partial replacement of dietary buffalo meat on the bone with chicken carcass improves serum antioxidant profile of zoo-housed Indian leopards (Panthera pardus fusca).

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 T1 were fed normal zoo diet of BMB. On the basis of dry matter, 10% and 20% of BMB was replaced with CC in groups T2 and T3 , 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.

Serum prolactin and testosterone levels in captive and wild brown kiwi (Apteryx mantelli) during the prebreeding, breeding, and incubation periods.

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.

SEASONAL VARIATION OF SERUM 25-HYDROXY-VITAMIN D IN TWO CAPTIVE EASTERN BLACK RHINOCEROS ( DICEROS BICORNIS MICHAELI) HOUSED IN A NORTH AMERICAN ZOO.

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 D3 (25OHD3; 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 D3 was fed to both rhinos. This study correlates environmental ultraviolet (UV) index, dietary supplementation, and seasonal serum 25OHD levels to compare with known 25OHD3 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 D3 supplementation levels may have little effect for Diceros spp. in human care housed in northern latitudes.