Serum chemistry values for free-ranging ringed seals (Pusa hispida) in Svalbard

BACKGROUND: Diseases and abnormal physiologic conditions can alter the concentrations of enzymes, metabolites, minerals, and hormones in the blood of animals. The ringed seal (Pusa hispida) has been selected as a key species for environmental monitoring, but information on disease and health parameters for this species is scarce. OBJECTIVES: The aim of the study reported here was to obtain serum chemistry reference intervals for free-ranging ringed seals in Svalbard, and then to evaluate serum chemistry values in relation to age, body condition, and sex. METHODS: Blood samples were collected after death from ringed seals in Wijdefjorden and Billefjorden, Svalbard (2002-2003; n = 75). Serum was analyzed for 24 selected serum chemistry parameters (enzymes, protein, metabolites, minerals, and cortisol). RESULTS: Compared with younger or older animals, seals between 7 and 16 years of age had larger variations in the activities of alanine transaminase, aspartate transaminase, lactate dehydrogenase, and creatine kinase (CK). Animals classified as having low body condition status had more variation in the serum activity of these enzymes, compared with that in animals with higher condition scores. Serum cortisol concentration was higher in young animals (1-5 years) than in older animals. Serum CK activity was higher in males than in females. CONCLUSION: The data reported here may be useful in monitoring the health of ringed seals and for tracking the impact of environmental changes in the Arctic.     

Serum chemistry of free-ranging white whales (Delphinapterus leucas) in Svalbard

BACKGROUND: Abnormal physiological conditions and diseases can change the concentrations of enzymes, metabolites, and minerals in the body. Serum chemistry information may thus be indicative of a specific disease; interpretation of such information requires knowledge of serum chemistry reference intervals from a seemingly healthy population of the species. OBJECTIVE: The aim of this study was to obtain serum chemistry reference intervals for a population of white whales. METHODS: Blood samples were collected from 21 free-ranging white whales (beluga; Delphinapterus leucas). The whales were live-captured in nets during 1996-2001 in Storfjorden, Van Mijenfjorden, and Van Keulenfjorden (Svalbard, Norway). While the whales were briefly physically restrained, blood was collected from the caudal vein into vacuum tubes without anticoagulant. The blood was left to clot for 4-6 hours before serum was obtained by centrifugation. The serum samples were then kept at -20 degrees C until analysis. Enzymes (aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase [ALP], creatine kinase, lactate dehydrogenase [LDH], amylase, lipase), metabolites (urea, creatinine, bilirubin, cholesterol, triglycerides, nonesterified fatty acids, glucose), and minerals (calcium, phosphate, magnesium, sodium, potassium, chloride) were analyzed in an Advia 1650 System (Bayer, Tarrytown, NY, USA). Cortisol was analyzed in an Immulite One system (Diagnostic Products Corporation, Los Angeles, CA, USA). The major blood proteins (albumin and globulins) were separated by gel electrophoresis in a Beckman Paragon electrophoresis system (Beckman Coulter, Inc., Fullerton, CA, USA). RESULTS: Serum values for all analytes were reported as median and range, and reference intervals were calculated as 10-90th percentiles. Activities of ALP and LDH and cortisol concentration were higher, and protein and bilirubin concentrations were lower compared with those previously reported for white whales from Canada; remaining results were strikingly similar in these 2 white whale populations. CONCLUSIONS: These data provide valuable serum chemistry reference intervals for future health assessments of white whales in Svalbard and other white whale populations, as well as captive individuals.     

Hematology and serum clinical chemistry reference intervals for free‐ranging Scandinavian gray wolves (Canis lupus)

Background: Scandinavian free‐ranging wolves (Canis lupus) are endangered, such that laboratory data to assess their health status is increasingly important. Although wolves have been studied for decades, most biological information comes from captive animals. Objectives: The objective of the present study was to establish reference intervals for 30 clinical chemical and 8 hematologic analytes in Scandinavian free‐ranging wolves. Methods: All wolves were tracked and chemically immobilized from a helicopter before examination and blood sampling in the winter of 7 consecutive years (1998–2004). Seventy‐nine blood samples were collected from 57 gray wolves, including 24 juveniles (24 samples), 17 adult females (25 samples), and 16 adult males (30 samples). Whole blood and serum samples were stored at refrigeration temperature for 1–3 days before hematologic analyses and for 1–5 days before serum biochemical analyses. Reference intervals were calculated as 95% confidence intervals except for juveniles where the minimum and maximum values were used. Results: Significant differences were observed between adult and juvenile wolves for RBC parameters, alkaline phosphatase and amylase activities, and total protein, albumin, γ‐globulins, cholesterol, creatinine, calcium, chloride, magnesium, phosphate, and sodium concentrations. Conclusion: Compared with published reference values for captive wolves, reference intervals for free‐ranging wolves reflected exercise activity associated with capture (higher creatine kinase activity, higher glucose concentration), and differences in nutritional status (higher urea concentration).     

Clinical chemistry values and tissue enzyme activities in western barred bandicoots (Perameles bougainville)

BACKGROUND: The western barred bandicoot, Perameles bougainville, is an endangered Australian marsupial species whose survival is threatened by a papillomatosis and carcinomatosis syndrome. Investigations to characterize this syndrome would benefit from species-specific clinical chemistry data. OBJECTIVES: The purpose of this study was to determine plasma biochemical reference values and to determine enzyme activities in various tissues of P. bougainville. METHODS: Heparinized blood samples were collected by jugular venipuncture from 53 clinically healthy bandicoots of both sexes and at 3 geographic locations. Plasma was analyzed for routine clinical chemistry variables using an automated biochemistry analyzer. Tissues obtained following humane euthanasia of 3 bandicoots were analyzed for alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), creatine kinase (CK), alpha-amylase (AML), and gamma-glutamyltransferase (GGT) activities. RESULTS: Significant differences in the results were found for animals based on geographic location and sex; hence, results were expressed as minimum and maximum values. A population reference interval was calculated for AST activity (20-283 U/L). ALT was found mainly in the liver, with lower levels in cardiac and skeletal muscle and kidneys. AST was detectable in many tissues, including the heart, liver, kidneys, and central nervous system; CK was found in skeletal and cardiac muscle and central nervous system; AML was found in the pancreas; and GGT was found mainly in kidneys with lower levels in the intestines and pancreas. CONCLUSIONS: These findings will facilitate the interpretation of clinical chemistry results from P. bougainville and thereby inform population management and clinical decision-making.     

Plasma biochemical reference values in clinically healthy captive bearded dragons (Pogona vitticeps) and the effects of sex and season

Background: Bearded dragons are one of the most popular pet lizard species, and biochemical reference values are useful for health management of these reptiles. Objectives: The objectives of this study were to measure plasma biochemical values in healthy captive bearded dragons, determine reference values, and evaluate the effects of sex and season on the results. Methods: Blood samples were collected from 100 captive healthy bearded dragons in Tokyo during the summer and winter. Plasma biochemical measurements were performed using a dry‐slide automated biochemical analyzer. The data were then compared based on sex and season using 2‐way ANOVA. Results: Globulin, cholesterol, and calcium concentrations of females were higher in both summer and winter compared with the values obtained for males. Both males and females had higher uric acid concentrations in winter than in summer. When compared with males, females had a higher chloride concentration in summer and a higher total protein concentration and aspartate aminotransferase activity in winter. Potassium concentration in males was lower in winter than in summer, whereas in females cholesterol concentration was lower in winter than in summer. Conclusions: Biochemical values that differed based on sex and season in bearded dragons were similar to those in other lizards. These differences reflect physiologic differences in reproductive status in females and seasonal changes in temperature and hydration status. Plasma biochemical values established for bearded dragons in this study will be useful in the diagnostic assessment of captive animals.     

Radiological and osteological study of the pelvic limbs in free‐ranging capybaras (Hydrochoerus hydrochaeris)

This study aimed to describe the radiographic anatomy and osteology and to evaluate angular radiographic measurements—Norberg angle, inclination angle and anteversion angle—of the pelvic limbs in free‐ranging capybaras. Twenty cadavers of free‐ranging capybaras (Hydrochoerus hydrochaeris), including five adults and 15 subadults, were studied. Ventrodorsal, craniocaudal, dorsoplantar and mediolateral radiographic views of the pelvic limbs were obtained. The radiographic features were described together with bone samples. The hip bone (os coxae), shaped like an isosceles trapezoid, was elongated and narrow with the presence of an oblong obturator foramen, sagittal ilial wing and rectilinear ilial body. The femoral shaft was relatively straight, while the greater trochanter was projected above the femoral head. No sesamoid bones of the gastrocnemius and popliteus muscles were observed radiographically or for those animals used in gross macroscopy. The fibula was located lateral and parallel to the tibia. Eight tarsal bones, four metatarsal bones and three digits were identified. The mean radiographic measurements included Norberg angle of 125.9°; respective angles of femoral inclination by the Hauptman B and Tomlinson methods of 139.9 and 141°; anteversion angle of the femoral head and neck of 29.80°. The bones of the pelvic limbs in capybaras have several anatomical characters and radiological features that are shared with members of the caviomorph superfamily Cavioidea. The radiographic angles measured in this study help characterize the functional morphology of this species.     

Substrate use and postural behavior in free‐ranging snub‐nosed monkeys (Rhinopithecus bieti) in Yunnan

Extant colobines represent a predominantly arboreal primate radiation, but there are outliers such as some species of snub‐nosed monkeys (Rhinopithecus spp.) that utilize both arboreal and terrestrial substrates. We quantified the degree of terrestriality and the distribution of individuals across canopy levels, the extrinsic and intrinsic determinants and constraints of strata association as well as use of postures in relation to behavior and food sources in a wild population of Yunnan snub‐nosed monkeys (Rhinopithecus bieti) in the Samage Forest, China. The subjects spent 80.5% of the observation time in the trees and 19.5% on the ground. Ground use occurred most frequently in the context of travelling (61%) and less often in the context of feeding and resting (30% combined). Adult males frequented the ground significantly more often than other age‐sex classes. Age‐sex differences were also apparent in substrate preferences, which likely reflect body weight constraints. Adult males were significantly more associated with solid substrates and less with terminal branches than adult females and juveniles. Juveniles were seen more often on unstable ground (i.e. on thin and oblique branches) and in the upper canopy. Substrate use also varied with behavior: feeding was much more common in the upper stratum, outer canopy, on highly inclined and thin branches, as compared to resting. Severe weather also influenced canopy use in that tree crowns were used less often during periods of snowfall. The most frequently exhibited resting and feeding postures was sitting. Standing and suspensory postures were most often used in the context of feeding on lichens.     

Antimicrobial resistance genes present in the faecal microbiota of free‐living Galapagos tortoises (Chelonoidis porteri)

Antimicrobial resistance (AMR), encoded by plasmid‐mediated AMR genes (ARGs), is an increasing global public health threat. Wildlife play a fundamental role as sentinels, reservoirs and potential vectors of ARGs. For the first time in Galapagos, we have identified and quantified the presence of ARGs in free‐living giant tortoises (Chelonoidis porteri). We performed ARG analyses by quantitative PCR of faeces collected from the cloaca of 30 tortoises widely distributed across Santa Cruz Island. Validated samples (n = 28) were analysed by a panel of up to 21 different ARGs and all 28 tortoise samples were positive to one or more genes encoding resistance. Thirteen of 21 tested ARGs were present in at least one sample, and 10 tortoises (35.7%) had a multi‐resistant pattern. We recommend additional research so we may more fully understand resistance patterns across taxa and geographical locations throughout the Galapagos archipelago, and the implications of ARGs for the health of wildlife, domestic animals, and humans. In this study, we found 100% of sampled giant tortoises had ARGs present in their faeces, suggesting a large‐scale distribution of these genes within the archipelago.     

Prevalence of Brucella pinnipediae in healthy hooded seals (Cystophora cristata) from the North Atlantic Ocean and ringed seals (Phoca hispida) from Svalbard

Investigations for Brucella-infections were conducted in 29 hooded seals (Cystophora cristata) caught between Svalbard and Greenland (North Atlantic Ocean; Greenland Sea) autumn 2002, and from 20 ringed seals (Phoca hispida) caught in Billefjord, Svalbard, spring 2003. All animals were apparently healthy and were caught in their natural habitat. Bacteriology on tissue samples from ringed seals was negative, whereas Brucella sp. were recovered in tissues from 11 of the 29 hooded seals (38%), with the highest tissue prevalence in spleen (9/29) and lung lymph nodes (9/24). Anti-Brucella antibodies were detected in sera from 9 hooded seals (31%) (EDTA-modified Slow Agglutination test of Wright, Rose Bengal test, Complement Fixation Test, and Protein-A ELISA). The bacterial isolates all belonged to the genus Brucella according to classical biotyping and PCR analysis based on Insertion Sequence IS711, and were shown to be typical marine mammal strains, based on the occurrence of an IS711 element downstream of the bp26 gene. Their dependency on CO2 for growth, and the presence of one copy each of the omp2a and omp2b gene finally classified them as Brucella pinnipediae. Furthermore, all the hooded seal isolates showed an A+ M+ agglutination profile, which is different from the profile of reference seal strain 2/94 (harbour seal, Phoca vitulina). Thus, these results indicate that B. pinnipediae may contain different biovars. The present results suggest that infection with B. pinnipediae is enzootic in this population. Since the hooded seal is commercially hunted and consumed in Norway, the pathological impact of such infections and their zoonotic potential should be further addressed.     

Pharmacokinetics and bioavailability after intramuscular injection of the 5‐HT1A serotonin agonist R‐8‐hydroxy‐2‐(di‐n‐propylamino) tetralin (8‐OH‐DPAT) in domestic goats (Capra aegagrus hircus)

To determine the bioavailability and pharmacokinetic properties of the serotonin 5‐HT_1A receptor agonist R‐8‐OH‐DPAT in goats, and 0.1 mg kg^?1 R‐8‐OH‐DPAT hydrobromide was administered intramuscularly (i.m.) and intravenously (i.v.) to six goats in a two‐phase cross‐over design experiment. Venous blood samples were collected from the jugular vein 2, 5, 10, 15, 20, 30, 40 and 60 min following treatment and analysed by liquid chromatography tandem mass spectrometry. Bioavailability and pharmacokinetic parameters were determined by a one‐compartment analysis. Mean bioavailability of R‐8‐OH‐DPAT when injected i.m. was 66%. The mean volume of distribution in the central compartment was 1.47 L kg^?1. The mean plasma body clearance was 0.056 L kg^?1 min^?1. All goats injected i.v. and two of six goats injected i.m. showed signs of serotonin toxicity. In conclusion, R‐8‐OH‐DPAT is well absorbed following i.m. injection and the observed pharmacokinetics suggest that administration via dart is feasible. Administration of R‐8‐OH‐DPAT hydrobromide, at a dosage of 0.1 mg kg^?1, resulted in the observation of clinical signs of serotonin toxicity in the goats. It is suggested that dosages for the clinical use of the compound should be lower in order to achieve the desired clinical effect without causing serotonin toxicity.     

Comparison of plasma vitamin A and E,copper and zinc levels in free‐ranging and captive greater flamingos (Phoenicopterus roseus) and their relation to pododermatitis

Pododermatitis is a worldwide problem in captive flamingos. Studies in domestic poultry showed that nutrition is a possible influencing factor for pododermatitis. Vitamin A and E, copper and zinc levels were analysed in two different diets (diet 1 = in‐house mix and diet 2 = commercial diet) and in plasma of captive greater flamingos fed these diets and compared to those of free‐ranging greater flamingos. Results were analysed with respect to type and severity of foot lesions of the individuals from the different groups. Juvenile and subadult/adult captive flamingos on diet 1 showed various types and severities of foot lesions, whereas no foot lesions were found at the time of blood sampling in juvenile captive flamingos on diet 2. Juvenile captive flamingos on diet 1 had significantly lower plasma zinc levels than juvenile captive flamingos on diet 2 and juvenile free‐ranging flamingos; data were also lower than reference ranges for flamingos, poultry and cranes. There were no significant differences in plasma vitamin A, vitamin E, copper or zinc levels between animals with different types of foot lesions or with different severity scores. Shortly after the change to diet 2 (fed to juvenile captive flamingos that did not show any foot lesion), the flooring of the outdoor water pools was covered with fine granular sand. Because both factors (nutrition and flooring) were changed during the same evaluation period, it cannot be concluded which factor contributed in what extent to the reduction of foot lesions. While it is assumed that low plasma zinc levels identified in the group of juvenile captive flamingos on diet 1 were not directly responsible for foot lesions observed in these animals, they may have played a role in altering the skin integrity of the feet and predisposing them to pododermatitis.     

Successful eradication of the European rabbit (Oryctolagus cuniculus) and house mouse (Mus musculus) from the island of Selvagem Grande (Macaronesian archipelago), in the Eastern Atlantic

The Portuguese island of Selvagem Grande (Great Salvage) in Macaronesia is an important seabird breeding station in the eastern Atlantic. Significant populations of Cory's shearwater Calonectris diomedea (Scopoli, 1769), Bulwer's petrel Bulweria bulweria (Jardine & Selby, 1828) and little shearwater Puffinus assimilis baroli (Bonaparte, 1857) are present, and white-faced storm-petrel Pelagodroma marina (Latham, 1790) and Madeiran storm-petrel Oceanodroma castro (Harcourt, 1851) populations are of global significance. Selvagem Grande also provides diverse habitats for an extensive flora, including 11 endemic species. The 270-ha island was also inhabited by two alien invasive mammals: the European rabbit Oryctolagus cuniculus (Linnaeus, 1758) and the house mouse Mus musculus (Linnaeus, 1758). Both are known to have had adverse impacts on breeding seabirds and island vegetation. In 2002, the Natural Park of Madeira conducted a program using brodifacoum bait formulations aimed at rabbit and mouse eradication. Approximately 17 000 individual baiting points were established on a 12.5 × 12.5 m grid. Baits were also applied by hand “seeding” on steep slopes and cliffs where bait stations could not be placed. Rabbits were removed after a month. However, mice persisted for considerably longer and strategic bait applications against them continued for a further six months. Subsequent assessments by trapping, bait takes and systematic observation of signs over three years, has confirmed the removal of both alien invasive species. This paper presents information on these operations, on measures adopted to mitigate adverse impacts of the eradication program on important vertebrate non-target species, including Berthelot's pipit Anthus berthelotii Bolle, 1862 and a species of gecko Tarentola bischoffi Joger, 1984 and on the initial response of the island's ecosystem to the eradication of rabbits and mice.     

Age‐dependent baseline values of faecal cortisol metabolites in the American mink (Neovison vison) under semi‐natural housing conditions

The welfare of an animal is ensured if it is able to fully satisfy its essential species‐typical needs in all functional aspects of behaviour. In mink, stereotypies and apathy, internal and/or external injuries as well as increased susceptibility to disease have been known to occur as a result of chronic stress. The non‐invasive method of analysing faecal cortisol metabolites (FCM) allows conclusions to be drawn about the stress level in the respective housing system. The objective of this study is to find out how the cortisol metabolites content in the faecal changes with increasing age of the mink under semi‐natural housing conditions. Thus, 40 American mink (Neovison vison) were housed in two outdoor enclosures imitating natural conditions. Throughout the entire study (13th to 32nd week of life), faecal samples were collected to measure cortisol metabolites. No differences in FCM concentrations between the two outdoor enclosures were found. In the young mink lower, less fluctuating FCM levels were found than in older animals. After the first faecal collection in the 13th/14th week of life, the level of metabolites decreased slightly (p = 0.032; 17th/18th week). From the 22nd/23rd week onwards until the 30th/31st week, shortly before the animals were pelted, continuously increasing concentrations were then measured. Increasing FCM levels with advancing age of the animals are probably attributable to the onset of sexual maturity and/or the respective season. This has to be taken into account in future studies using this method for assessing welfare and when comparing different mink housing systems.