Identifying potential measures of stress and disturbance during a captive to wild African elephant reintegration.

There is increasing evidence of compromised welfare for elephants managed in captivity. Should such facilities eventually close, more elephants will need to be rehabilitated and reintegrated into the wild. The goal of such reintegration would be to restore any physical or psychological aspects of the elephant that may have been compromised in captivity, followed by introduction into a free-roaming system where they can interact with other elephants. However, to achieve this goal, the reintegration methods implemented need to be assessed to ensure that welfare remains the priority. The objective of this study was to test whether parameters generally associated with stress and disturbance in African elephants, respond to changes in potentially stressful environmental conditions, assessed at multiple temporal scales ranging from minutes to months. The main changes in environmental conditions that were investigated included the different phases of reintegration of a group of elephants from captivity into the wild. Stress and disturbance related parameters used for comparisons included physiological responses, namely the extent of temporal gland secretions (eTGS) and faecal glucocorticoid metabolite (fGCM) levels as well as behavioural responses, namely the display of stereotype and stress-related behaviours. Results showed that eTGS significantly increased during the initial release of the elephants compared to when in captivity. Stereotypic behaviours were only recorded during the captive phase and immediately ceased after release. Faecal GCM levels spiked in the first year after release before decreasing back to pre-reintegration levels during the third year. These findings indicate that fGCM levels, the eTGS and disturbance related behaviours all proved effective in explaining the changes in stress and disturbance experienced by elephants during the initial years after being reintegrated from captivity into the wild.

Factors affecting captive female giraffe stress response: Male presence, small enclosure, and low temperature.

To improve animal welfare based on suitable social housing conditions, it is important to understand the factors that trigger high-stress responses. Wild giraffes live in a fission-fusion society and males and females are rarely in the same herd for a long period. The captive condition of belonging to a herd with the same individuals for months or years is uncommon in nature. To understand the effect of male presence on female stress levels, fecal glucocorticoid metabolite (fGCM) levels and social interactions in two captive female giraffes were investigated. Additionally, the effect of enclosure size and temperature on fGCM level and social interactions were examined. The results showed no significant difference in the fGCM levels of females based on male presence. The frequency of agonistic behavior by the dominant female toward the subordinate female was significantly increased when a male was present. The subordinate female was significantly less likely to approach the dominant female and showed decreased affiliative and agonistic interactions toward the dominant female when a male was present. The frequencies of agonistic interactions between females were higher in the small enclosure regardless of male presence. Low temperature triggered higher fGCM levels and increased agonistic interaction in an aged female. The findings of this study suggest that these multiple factors should be considered individually to promote the welfare of captive giraffes.

Simultaneous determination of selected catechins and pyrogallol in deer intoxications by HPLC-MS/MS.

A selective and sensitive analytical method for the determination of selected catechins (catechin, epicatechin, gallocatechin, epigallocatechin) and pyrogallol in biological matrices by HPLC-MS/MS was developed. The utilized sample preparation technique was a two-stage liquid-liquid extraction using ethyl acetate. The HPLC-system was equipped with a Phenomenex Luna Pentafluorophenyl Column (150 × 2 mm, 5 µm) and operated with an acetonitrile-water gradient as a mobile phase system. Detection was performed with a 3200 Q Trap mass spectrometer. For analysis the mass spectrometer was used in the MRM-mode with negative ionization. The method validation was performed with serum as matrix. The selectivity of the method as well as the linearity of calibration was successfully proven for all analytes. The limits of quantification were between 5.3 and 11.2 ng/mL and the recovery rates were above 50 % for all analytes. Results from the samples of three deer poisoning cases demonstrated that the developed HPLC-MS/MS method is applicable to real biological samples.

Reference intervals for Cu, Mg, and Zn in captive gyrfalcons and other falcon species in the United Arab Emirates.

Falconry is a long-standing tradition in the United Arab Emirates (UAE), and in 2016, falconry was included by UNESCO in the list of intangible cultural heritage of humanity. The health and wellness of falconry raptors is a priority for the local culture; plasma chemistry analysis plays an important role in monitoring, assessing, and managing diseases in avian patients. Imbalances of Cu, Mg, and Zn have been linked to several diseases in avian species; therefore, determining the reference intervals (RIs) of these minerals has important implications in the clinical management of falcons. We determined the RIs in plasma for Cu, Mg, and Zn in captive (falconry-managed) falcons according to the Quality Assurance and Laboratory Standards Committee of the American Society for Veterinary Clinical Pathology, and the Clinical Laboratory Standards Institute guidelines. Blood chemistry analyses were carried out on 252 clinically healthy falcons examined at the Dubai Falcon Hospital in the UAE: 124 gyrfalcons (Falco rusticolus) and 128 falcons of other species. We observed no significant variation in RIs of Cu (1.5-7.0 µmol/L), Mg (0.49-0.78 mmol/L), or Zn (11.8-34.1 µmol/L) based on different species or sex.

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.

Aflatoxin B1 and Sterigmatocystin Binding Potential of Non-Lactobacillus LAB Strains.

Research on the ability of lactic acid bacteria (LAB) to bind aflatoxin B1 (AFB1) has mostly been focusing on lactobacilli and bifidobacteria. In this study, the AFB1 binding capacities of 20 Enterococcus strains belonging to E. casseliflavus, E. faecalis, E. faecium, E. hirae, E. lactis, and E. mundtii, 24 Pediococcus strains belonging to species P. acidilactici, P. lolii, P. pentosaceus, and P. stilesii, one strain of Lactococcus formosensis and L.garviae, and 3 strains of Weissella soli were investigated in MRS broth at 37 °C at 0.2 µg/mL mycotoxin concentration. According to our results, among non-lactobacilli LAB, the genera with the best AFB1 binding abilities were genus Pediococcus, with a maximum binding percentage of 7.6% by P. acidilactici OR83, followed by genus Lactococcus. For AFB1 bio-detoxification purposes, beside lactobacilli, pediococci can also be chosen, but it is important to select a strain with better binding properties than the average value of its genus. Five Pediococcus strains have been selected to compare their sterigmatocystin (ST) binding abilities to AFB1 binding, and a 2-3-fold difference was obtained similar to previous findings for lactobacilli. The best strain was P. acidilactici OR83 with 18% ST binding capacity. This is the first report on ST binding capabilities of non-Lactobacillus LAB strains.

Gut microbiota and fecal metabolites in captive and wild North China leopard (Panthera pardus japonensis) by comparsion using 16 s rRNA gene sequencing and LC/MS-based metabolomics.

BACKGROUND: Gut microbes significantly contribute to nutrient digestion and absorption, intestinal health and immunity, and are essential for the survival and environmental adaptation of wild animals. However, there are few studies on the gut microbiota of captive and wild North China leopard (Panthera pardus japonensis). RESULTS: A total of 10 mainly bacterial phyla were identified in the fecal microbiota of North China leopard, Lachnoclostridium (p = 0.003), Peptoclostridium (p = 0.005), Bacteroides (p = 0.008), Fusobacterium (p = 0.017) and Collinsella (p = 0.019) were significantly higher than those of wild North China leopard. Distinct differences in the fecal metabolic phenotypes of captive and wild North China leopard were found, such as content of l-methionine, n-acetyl-l-tyrosine, pentadecanoic acid and oleic acid. Differentially abundant gut microbes were associated with fecal metabolites, especially the bacteria in Firmicutes and Bacteroidetes, involved in the metabolism of N-acetyl-L-alanine and D-quinovose. CONCLUSION: This study reports for the first time the differences in gut microbiota abundance between captive and wild North China leopard, as well as significant differences in fecal metabolic phenotypes between two groups.

Inflammatory and oxidative status in European captive black rhinoceroses: A link with Iron Overload Disorder?

Iron Overload Disorder (IOD) is a syndrome developed by captive browsing rhinoceroses like black rhinoceroses (Diceros bicornis), in which hemosiderosis develops in vital organs while free iron accumulates in the body, potentially predisposing to various secondary diseases. Captive grazing species like white rhinoceroses (Ceratotherium simum) do not seem to be affected. The authors hypothesized that inflammation and oxidative stress may be implicated in the pathogenesis of IOD in captive black rhinoceroses, making this syndrome a potential common denominator to various diseases described in captivity in this species. In this prospective study, 15 black (BR) and 29 white rhinoceroses (WR) originating from 22 European zoos were blood-sampled and compared for their iron status (serum iron), liver/muscle biochemical parameters (AST, GGT, cholesterol), inflammatory status (total proteins, protein electrophoresis) and oxidative stress markers (SOD, GPX, dROMs). Results showed higher serum iron and liver enzyme levels in black rhinoceroses (P < 0.01), as well as higher dROMs (P < 0.01) and a trend for higher GPX (P = 0.06) levels. The albumin/globulin ratio was lower in black rhinoceroses (P < 0.05) due to higher α2-globulin levels (P < 0.001). The present study suggests a higher inflammatory and oxidative profile in captive BR than in WR, possibly in relation to iron status. This could be either a consequence or a cause of iron accumulation. Further investigations are needed to assess the prognostic value of the inflammatory and oxidative markers in captive black rhinoceroses, particularly for evaluating the impact of reduced-iron and antioxidant-supplemented diets.

Evaluation of vitamin D3 metabolites in Callithrix jacchus (common marmoset).

Vitamin D3 (cholecalciferol) is endogenously produced in the skin of primates when exposed to the appropriate wavelengths of ultraviolet light (UV-B). Common marmosets (Callithrix jacchus) maintained indoors require dietary provision of vitamin D3 due to lack of sunlight exposure. The minimum dietary vitamin D3 requirement and the maximum amount of vitamin D3 that can be metabolized by marmosets is unknown. Observations of metabolic bone disease and gastrointestinal malabsorption have led to wide variation in dietary vitamin D3 provision amongst research institutions, with resulting variation in circulating 25-hydroxyvitamin D3 (25(OH)D3 ), the accepted marker for vitamin D sufficiency/deficiency. Multiple studies have reported serum 25(OH)D3 in captive marmosets, but 25(OH)D3 is not the final product of vitamin D3 metabolism. In addition to serum 25(OH)D3, we measured the most physiologically active metabolite, 1,25-dihydroxyvitamin D3 (1,25(OH)2 D3 ), and the less well understood metabolite, 24,25-dihydroxyvitamin D3 (24,25(OH)2 D3 ) to characterize the marmoset's ability to metabolize dietary vitamin D3 . We present vitamin D3 metabolite and related serum chemistry value colony reference ranges in marmosets provided diets with 26,367 (Colony A, N = 113) or 8,888 (Colony B, N = 52) international units (IU) of dietary vitamin D3 per kilogram of dry matter. Colony A marmosets had higher serum 25(OH)D3 (426 ng/ml [SD 200] vs. 215 ng/ml [SD 113]) and 24,25(OH)2 D3 (53 ng/ml [SD 35] vs. 7 ng/ml [SD 5]). There was no difference in serum 1,25(OH)2 D3 between the colonies. Serum 1,25(OH)2 D3 increased and 25(OH)D3 decreased with age, but the effect was weak. Marmosets tightly regulate metabolism of dietary vitamin D3 into the active metabolite 1,25(OH)2 D3 ; excess 25(OH)D3 is metabolized into 24,25(OH)2 D3 . This ability explains the tolerance of high levels of dietary vitamin D3 by marmosets, however, our data suggest that these high dietary levels are not required.

THE VITAMIN D STATUS OF ASIAN ELEPHANTS (ELEPHAS MAXIMUS) MANAGED IN A NORTHERN TEMPERATE CLIMATE.

Knowledge about the normal metabolism and involvement of vitamin D in elephant calcium homeostasis is essential to understanding the possible role of vitamin D in Asian elephant (Elephas maximus) health, as well as to informing accurate diet formulation. This study provides an evaluation of analytes involved in vitamin D metabolism, in conjunction with dietary intake and ultraviolet light (UV) exposure, in Asian elephants managed in a northern temperate climate. Once monthly, for a total of 12 mo, serum from six adult Asian elephants was analyzed for 25-hydroxyvitamin D [25(OH)D], 24,25-dihydroxyvitamin D [24,25(OH)2D], 1,25-dihydroxyvitamin D [1,25(OH)2D], parathyroid hormone (PTH), total calcium (Ca), ionized calcium (iCa), phosphorus (P), and magnesium (Mg). The diet was analyzed monthly for vitamin D, Ca, and P. Monthly average vitamin D-weighted UV daily sums were determined to gauge average UV light exposure within the vitamin D action spectrum. No serum or diet parameters were affected by time or season. Average serum 25(OH)D2 was 7.02 ± 0.85 ng/ml. 25(OH)D3 levels were nondetectable in all samples despite supplementation of the diet with recommended levels of vitamin D3, and UV exposure was at sufficient levels for cutaneous vitamin D synthesis for 6 mo of the year. Levels of 24,25(OH)2D averaged 31.7% higher than 25(OH)D, and average 1,25(OH)2D2 was 11.24 ± 1.04 pg/ml. Values for PTH, Ca, iCa, P, and Mg were within expected ranges for Asian elephants. The information gained from this research expands the knowledge base for these analytes, evaluates 24,25-dihydroxyvitamin D for the first time, and provides new information regarding vitamin D metabolism and test interpretation in the Asian elephant.

PHARMACOKINETICS OF ORAL MAVACOXIB IN CARIBBEAN FLAMINGOS (PHOENICOPTERUS RUBER RUBER).

Mavacoxib is a selective cyclooxygenase-2 nonsteroidal anti-inflammatory drug that has been used for management of osteoarthritis and other inflammatory conditions in dogs. The main advantage of mavacoxib over other nonsteroidal anti-inflammatory drugs is its longer plasma half-life, leading to decreased dosing frequency. This study determined the pharmacokinetics of mavacoxib in Caribbean flamingos (Phoenicopterus ruber ruber) after a single-dose oral administration of 6 mg/kg (n = 6). Plasma mavacoxib concentrations were determined using liquid chromatography with mass spectrometry, and pharmacokinetic analysis was performed using noncompartmental methods. Mean peak plasma concentration (Cmax) was (mean; range) 2.97 (2.19--4.06) µg/ml; mean time to peak plasma concentration (Tmax) was 18.68 (4.00-48.00) hr; mean area under the curve (AUC) was 455 (292-637) hr * µg/ml; and mean terminal half-life (T1/2) was 74.47 (49.57-161.43) hr. Based on the results of this study, mavacoxib dosed at 6 mg/kg orally in Caribbean flamingos reaches plasma concentrations above the therapeutic concentration established for dogs, but further studies are needed to determine appropriate dosing recommendations in flamingos.

Longitudinal fecal hormone monitoring of adrenocortical function in zoo housed fishing cats (Prionailurus viverrinus) during institutional transfers and breeding introductions.

The ex situ population of fishing cats (Prionailurus viverrinus) has become increasingly important for the conservation of this species. Unfortunately, captivity-induced stress is a concern and potential factor for lack of breeding success in this small felid, resulting in an unsustainable population. The objectives of this study were to: 1) validate an enzyme immunoassay for monitoring of fecal glucocorticoid metabolite (FGM) concentrations in the fishing cat; 2) identify potential exogenous stressors in the captive environment; 3) pinpoint management techniques that may lower FGM concentrations; and 4) determine if FGM concentrations are related to breeding success. Through a successful adrenocorticotrophic hormone challenge and additional laboratory methods, a cortisol enzyme immunoassay was validated as an effective tool for detecting FGM in this species. Between 2010 and 2013, longitudinal FGM monitoring was conducted in 26 fishing cats in the North American Species Survival Plan®. Exogenous stressors that elevated FGM concentrations included: chemical immobilizations; permanent transfers between facilities; construction; facility events; and fights/aggression among breeding pairs. Management factors that lowered FGM concentrations included: increased animal-keeper interaction through formal training; and providing indoor, off-exhibit, holding areas. In addition, social housing of individuals (either established breeding pairs or same sex pairs) decreased FGM concentrations. Individuals with breeding success (defined as observed copulations during the study period) also had lower FGM concentrations than unsuccessful individuals. Findings indicate that management factors play a role in lowering glucocorticoid (stress) levels in fishing cats, which may ultimately affect breeding success in the ex situ population.

ANALYSIS OF FATTY ACID PROFILES FOR EASTERN BOX (TERRAPENE CAROLINA CAROLINA) AND COMMON SNAPPING (CHELYDRA SERPENTINE) TURTLES IN WILD AND MANAGED CARE ENVIRONMENTS.

The housing of wild animals in managed care facilities requires attention to all aspects of husbandry. Diets of wild animals often differ in composition, consistency, and quantity when compared with those in managed care settings including zoos, rehabilitation facilities, and aquaria. It was hypothesized that dietary differences from wild versus managed care would be reflected in data of circulating fatty acids based on previous studies. The current study examined the effect of species and environment on fatty acid concentrations in two omnivorous species of chelonians: Eastern box turtles, Terrapene carolina carolina, and common snapping turtles, Chelydra serpentina, located in the wild and managed care. Whole blood was collected and placed on spot cards for analysis of 26 fatty acids in a total lipid fatty acid profile. The present research indicated that Eastern box turtles have significantly (P < 0.05) higher percentages of linoleic acid (18:2n6), eicosadienoic acid (20:2n6), and mead acid (20:3n9). Common snapping turtles have significantly (P < 0.05) higher percentages of myristic acid (14:0), dihomo-γ-linolenic acid (20:3n6), erucic acid (22:1n9), and n-6 docosapentaenoic acid (22:5n6). Environmental effects also were noted; wild turtles had higher percentages of α-linolenic acid (18:3n3), arachidic acid (20:0), eicosadienoic acid (20:2n6), and eicosatrienoic acid (20:3n3) (P < 0.05), whereas n-6 docosapentaenoic acid (22:5n6) was higher for the managed care group. Eicosadienoic acid (20:2n6), behenic acid; 22:0), adrenic acid (22:4n6), n-6 docosapentaenoic acid (22:5n6), and nervonic acid (24:1) were significantly different (P < 0.05) in species-environment interactions without any noted species or environment patterns. Fatty acids are useful for many important biological functions including proper immune system regulation, and therefore, the present research provides medically relevant data for reptile diagnostics. This research may help further improve diets of all chelonians kept in managed care, regardless of species.

Characterization and correlations of behavioral and adrenocortical activities of zoo-housed lesser anteaters (Tamandua tetradactyla).

We characterized behavioral and adrenocortical activities of Tamandua tetradactyla under human care driven by the hypothesis that they vary between males and females. We also assessed the potential association between natural or abnormal behaviors and adrenocortical activity. We kept females and males T. tetradactyla in individual, contiguous enclosures at Córdoba Zoo (Argentina), under natural photoperiod and temperature. During 29 consecutive days we monitored the animals' behavior by recording their activity pattern every 5 min using infrared cameras (8352 records/individual). We collected all feces and measured fecal glucocorticoid metabolites (FGM) with an 11-oxoaetiocholanolone enzyme immunoassay. We found individual differences in all behavioral variables. We detected that females exhibited lower total activity than males (23.8 ± 0.2% and 32.3 ± 0.3%, respectively; p = .005). Females were more active at night and males during the day (p < .05) and exhibited less abnormal behaviors than males (p = .05). Although we did not find sex-related differences for average FGM, we detected individual differences (p < .0001). We found that daily FGM showed negative (-0.39) and positive (0.38) correlations with natural and abnormal behaviors, respectively (p < .0001). Thus, we consider that individual input and sex are factors to be considered in stress responses of the species in captivity. Natural and abnormal behaviors may demand different levels of adrenocortical activity. Our findings may prove useful as normative data for ex situ management of conservation programs.

Feathers of Humboldt penguin are suitable bioindicators of Rare Earth Elements.

Rare earth elements (REEs), also called lanthanides, are emerging contaminants worldwide, due to their unique physical and chemical characteristics that make them essential in a variety of industrial applications. However, there is still a gap in the knowledge of occurrence and accumulation of REEs in biota, and no investigations have yet been performed in penguin feathers, which have already been widely utilized as a non-invasive tool for the biomonitoring of trace elements. The concentrations of 16 REEs were investigated in a colony of Humboldt penguins (Spheniscus humboldti) housed at the Acquario di Cattolica (Italy). Multielement determination of REEs was performed by an Inductively Coupled Plasma-Mass Spectrometer after a microwave digestion of feathers. As this colony lives indoors in a controlled environment, it was the ideal choice for studying lanthanide occurrence in penguin feathers. Since there is a strict link between metal levels in feathers and the diet of penguins, their food (capelin) was also tested for REEs. Chondrite normalized values revealed the same pattern for REEs in feathers and fish, but REE concentrations were an order of magnitude higher in penguin feathers, demonstrating the suitability of this tissue as a bioindicator of REEs.

SURVEY OF ANTITUBERCULOSIS DRUG ADMINISTRATION AND ADVERSE EFFECTS IN ELEPHANTS IN NORTH AMERICA.

Tuberculosis, caused by Mycobacterium tuberculosis, is a disease causing morbidity and mortality in captive elephants (Elephas maximus and Loxodonta africana) as well as free-ranging individuals. Elephants in North America diagnosed with tuberculosis are often treated with antituberculosis drugs, unlike livestock species, which has necessitated the development of treatment guidelines adapted from recommendations for humans. There are few published reports describing empirical treatment, which may be complicated by poor patient compliance, interruptions in drug administration, and adverse effects. A survey of elephants in North America was conducted to compile information on treatment protocols, including drugs, dosages, routes of administration, serum drug concentrations, and adverse effects of antituberculosis treatment. Responses were received regarding 182 elephants, 12 of which were treated prophylactically or therapeutically with antituberculosis drugs. Treatment protocols varied among elephants, and included various combinations of isoniazid, rifampin, pyrazinamide, ethambutol, enrofloxacin, levofloxacin, and ethionamide. Serum drug concentrations also varied considerably among and within individuals. Facility staff reported 5 elephants (out of 7 treated elephants with responses) that exhibited clinical signs that may have been associated with antituberculosis drugs or treatment procedures. Anorexia, decreased water intake, constipation, depression, ataxia, limb paresis, and tremors were among the signs observed. Most adverse effects were reported to be moderate or severe, resulting in interruption of the treatment. The results from this survey provide veterinarians and elephant managers with valuable historical data to make informed clinical management decisions regarding antituberculosis therapy in elephants.

Humboldt penguins' feathers as bioindicators of metal exposure.

Avian feathers have the potential to accumulate trace elements originating from contaminated food and polluted environments. In fact, in feathers, metals bind to keratin, a sulphur-containing protein for which several metals have a strong affinity. Here, the concentrations of 18 essential and non-essential elements were investigated in a Humboldt penguin (Spheniscus humboldti) colony housed at the Acquario di Cattolica (Italy). This species is listed as vulnerable in the Red List of the International Union for Conservation of Nature. According to the literature, there is usually a link between metal levels in the diet of birds and levels detected in their feathers. Thus, metals were also determined in the penguins' food (capelin, Mallotus villosus). We hypothesize that the controlled conditions in which birds are kept in captivity, and the homogeneous diet that they follow could allow a better understanding of metal bioaccumulation (such as mercury) or bio-dilution (such as arsenic) in the marine food chain, indicated by penguins' feathers. Moreover, comparisons with our previous investigations performed on an ex-situ African penguin (Spheniscus demersus) colony suggest that penguins living indoors have lower body burden of metals than those living outdoors. Indeed, environmental contaminants usually found in areas subjected to anthropogenic impact, where zoos and aquaria are often located, are not accumulated to levels of concern.

COMPARISON OF TWO IODINE QUANTIFICATION METHODS IN AN ARTIFICIAL SEAWATER SYSTEM HOUSING WHITE-SPOTTED BAMBOO SHARKS ( CHILOSCYLLIUM PLAGIOSUM).

Iodine is an essential micronutrient for elasmobranchs in order to prevent goiter. Preventing goiter requires bioavailable iodide: either oral iodide or maintaining adequate aquarium water iodide concentrations. The objective of this study was to determine how oral and water supplementation affected iodine (I2) and iodide (I-) concentrations in artificial seawater aquaria housing captive white-spotted bamboo sharks ( Chiloscyllium plagiosum). Daily water samples were collected and free iodine (I2) was determined using ultraviolet-absorbance spectrophotometry (a relatively simple in-house assay) and total iodide (I-) via liquid chromatography (a more time- and expertise-intense quantification method) to learn the effects of supplementation. One water system received iodine and iodide supplementation in the form of 5% Lugol's iodine solution added directly to the water, while a second water system received no supplementation. In addition, one tank of sharks in each water system received oral iodide supplementation. Results indicated that oral supplementation provides greater increases in water concentrations of bioavailable iodide (I-) than direct water supplementation. In addition, the chromatographic results suggested that iodide is present in higher concentrations in the systems not receiving water supplementation. Increased iodide concentrations were detected in water samples after water changes and after oral iodide supplementation was administered, but total iodine (I2) concentration changes were not detectable within the same time frame.

Metals in Feathers of African Penguins (Spheniscus demersus): Considerations for the Welfare and Management of Seabirds Under Human Care.

Bird feathers have been proven to be reliable indicators of metal exposure originating from contaminated food and polluted environments. The concentrations of 15 essential and non-essential metals were investigated in African penguins (Spheniscus demersus) feathers from a Northwestern Italian zoological facility. These birds are exclusively fed with herring from the northeast Atlantic Ocean. Certain elements, such as Hg and Cd, reflected the bioaccumulation phenomena that occur through the marine food chain. The levels of Cr, Mn, and Ni were comparable to those registered in feathers of birds living in polluted areas. These results are important for comparative studies regarding the health, nutrition and welfare of endangered seabirds kept under human care.

One year daily changes in fecal sexual steroids of two captive female cheetahs (Acinonyx jubatus) in Italy.

The present study evaluated changes of fecal sexual steroids in two female cheetahs (Geijsha and Duchessa) in Northern Italy throughout one year. Wet feces were collected daily from two sibling animals of the same age, housed with conspecific males and managed in the same conditions, and estrogens and progestogens concentrations were analyzed by radioimmunoassay (RIA). Evidence of ovarian activity based on regular fluctuation in estrogen excretion was demonstrated in both females. None of the animals was continuously cycling, as follicular activity was interrupted by anestrous periods, during the spring and early winter. No significant increases of progestogens were recorded after the estrogen peaks, indicating that induced or spontaneous ovulations did not occur during the observation period. The wavelet decomposition evidenced the temporal pattern of ovarian activity in the two females, underlying throughout the year a more pronounced rhythmical ovarian estrogenic activity in Geijsha than in Duchessa. However, this statistical approach had a smoothing effect in depicting the hormonal patterns and the number of follicular phases might be lower than that revealed by the iterative method. In this study, RIA on wet feces performed very well to determine sexual steroid concentrations, and an ovarian activity interrupted by anestrous periods along the year in captive cheetahs co-housed in a small group was demonstrated. More information on estrous behavior of captive cheetahs were obtained in this study, but the effects of husbandry and management conditions on natural reproductive physiology of this species remain to elucidate.