Baseline and stress-induced corticosterone levels are heritable and genetically correlated in a barn owl population.

The hypothalamic-pituitary-adrenal (HPA) axis is responsible for the regulation of corticosterone, a hormone that is essential in the mediation of energy allocation and physiological stress. As a continuous source of challenge and stress for organisms, the environment has promoted the evolution of physiological adaptations and led to a great variation in corticosterone profiles within or among individuals, populations and species. In order to evolve via natural selection, corticosterone levels do not only depend on the strength of selection exerted on them, but also on the extent to which the regulation of corticosterone is heritable. Nevertheless, the heritability of corticosterone profiles in wild populations is still poorly understood. In this study, we estimated the heritability of baseline and stress-induced corticosterone levels in barn owl (Tyto alba) nestlings from 8 years of data, using a multivariate animal model based on a behavioural pedigree. We found that baseline and stress-induced corticosterone levels are strongly genetically correlated (r = 0.68-0.80) and that the heritability of stress-induced corticosterone levels (h2 = 0.24-0.33) was moderate and similar to the heritability of baseline corticosterone levels (h2 = 0.19-0.30). These findings suggest that the regulation of stress-induced corticosterone and baseline levels evolves at a similar pace when selection acts with the same intensity on both traits and that contrary to previous studies, the evolution of baseline and stress-induced level is interdependent in barn owls, as they may be strongly genetically correlated.

Effects of Isoflurane Anesthesia on the Hematologic Values of Rehabilitated Wild Owls.

Isoflurane anesthesia is commonly used for owls when they are being rehabilitated to minimize stress during treatments and procedures, as well as to ensure caretaker safety. However, the effects of isoflurane anesthesia on the hematologic response of owls are not known. To investigate the effects of isoflurane anesthesia on the hematology of owls, 3 phases of investigation were performed on the subject animals: 1) single, short manual- versus single, short isoflurane-restraint episodes (n = 12; 38%); 2) a single, prolonged isoflurane episode (n = 10; 31%); and 3) serial, short isoflurane episodes (n = 10; 31%). All owls were classified as adult, and the sex for most individuals was unknown. Twelve owls (38%) were included in phase 1: 5 great horned owls (Bubo virginianus; 42%), 2 eastern screech owls (Megascops asio; 17%), and 5 barred owls (Strix varia; 42%). A separate cohort of 10 novel owls (31%) were selected for inclusion in both phases 2 and 3: 4 great horned owls (40%), 2 eastern screech owls (20%), 2 barred owls (20%), 1 barn owl (Tyto alba; 10%), and 1 snowy owl (Bubo scandiacus; 10%). For each anesthetic episode, blood was collected within 3 minutes of capture and in 15-minute intervals according to the duration of the procedure. Phase 2 had additional blood collections with the patient awake at 2 and 24 hours after time 0 blood collection, whereas phase 3 had an additional blood collection at 24 hours after time 0 blood collection. Hematologic analyses included packed cell volume, total solids, total white blood cell count, heterophil to lymphocyte ratio, and absolute heterophil, lymphocyte, monocyte, eosinophil, and basophil counts. Total white blood cell count decreased significantly during phase 1; packed cell volume decreased significantly during phases 2 and 3; total solids decreased significantly in phase 2; phase 2 demonstrated a lymphopenia with a concurrent decrease in the heterophil to lymphocyte ratio; and phase 3 demonstrated a heteropenia and significant changes in the eosinophil count. All hematologic changes noted in the study were within appropriate reference intervals for the owls but do suggest that there are physiologic consequences of restraining and anesthetizing these avian patients.

Detecting Burrowing Owl Bloodmeals in Pulex irritans (Siphonaptera: Pulicidae).

Pulex irritans L. is a cosmopolitan flea species that infests a wide variety of hosts. In North America it generally parasitizes large wild mammals, but in the Pacific Northwest an association has emerged between P. irritans and the western burrowing owl (Athene cunicularia hypugaea). While investigators have recognized this association for decades, it has not been clear if P. irritans feeds on burrowing owls, or if the owls serve exclusively as phoretic hosts. Here we describe using a real-time assay that was originally developed to identify bloodmeals in Ugandan cat fleas (Ctenocephalides felis Bouché) to detect burrowing owl DNA in P. irritans collected from burrowing owls in southern Idaho. Of 50 fleas tested, 12 had no detectable vertebrate bloodmeal. The remaining 38 (76%) contained burrowing owl DNA. The assay did not detect vertebrate DNA in unfed fleas exposed to owl or mouse pelts and is therefore unlikely to detect DNA in fleas from vertebrates that have served exclusively as phoretic hosts. We conclude that P. irritans feeds on burrowing owls. We discuss the potential implications of this finding for burrowing owl conservation and enzootic plague dynamics.

EXPLORATORY PLASMA BIOCHEMISTRY REFERENCE INTERVALS FOR URAL OWLS (STRIX URALENSIS, PALLAS 1771) FROM THE AUSTRIAN REINTRODUCTION PROJECT.

The Ural owl (Strix uralensis) is the biggest forest-living owl in Austria; however, it became extinct in Austria through poaching and habitat loss more than half a century ago. The birds examined in the present study were breeding pairs from the reintroduction project with the aim of determining exploratory plasma biochemistry reference intervals in Ural owls and evaluating the amount of biological variation between seasons, sexes, and ages. A total of 45 birds were sampled, including 13 adult males, 14 adult females, and 18 juvenile birds. Remarkably, almost all of the analytes showed significant differences between the subgroups, primarily between seasons, followed by age and sex. Only creatinkinase, glucose, lactatdehydrogenase, and triglycerides did not show any significant variations. Despite partitioning of reference values into subgroups according to biological variation diminishing the number of reference individuals in the respective groups, the resulting smaller reference intervals will improve medical assessment. The results of the present study once again demonstrate that significant seasonal fluctuations must be expected and considered in the interpretation. It can be assumed that these differences are probably even greater in free-range birds with considerable changes in food quantity and quality during and between years.

Agricultural land use and human presence around breeding sites increase stress-hormone levels and decrease body mass in barn owl nestlings.

Human activities can have a suite of positive and negative effects on animals and thus can affect various life history parameters. Human presence and agricultural practice can be perceived as stressors to which animals react with the secretion of glucocorticoids. The acute short-term secretion of glucocorticoids is considered beneficial and helps an animal to redirect energy and behaviour to cope with a critical situation. However, a long-term increase of glucocorticoids can impair e.g. growth and immune functions. We investigated how nestling barn owls (Tyto alba) are affected by the surrounding landscape and by human activities around their nest sites. We studied these effects on two response levels: (a) the physiological level of the hypothalamus-pituitary-adrenal axis, represented by baseline concentrations of corticosterone and the concentration attained by a standardized stressor; (b) fitness parameters: growth of the nestlings and breeding performance. Nestlings growing up in intensively cultivated areas showed increased baseline corticosterone levels late in the season and had an increased corticosterone release after a stressful event, while their body mass was decreased. Nestlings experiencing frequent anthropogenic disturbance had elevated baseline corticosterone levels, an increased corticosterone stress response and a lower body mass. Finally, breeding performance was better in structurally more diverse landscapes. In conclusion, anthropogenic disturbance affects offspring quality rather than quantity, whereas agricultural practices affect both life history traits.

PHARMACOKINETIC PROPERTIES OF A SINGLE ADMINISTRATION OF ORAL GABAPENTIN IN THE GREAT HORNED OWL (BUBO VIRGINIANUS).

Gabapentin (1-[aminomethyl] cyclohexane acetic acid) is a γ-aminobutyric acid analogue that has been shown to be efficacious for neuropathic pain control in humans. Plasma gabapentin concentrations >2 µg/ml are considered effective in treating epilepsy in humans and are suggested to provide analgesia for neuropathic pain. This study investigated the pharmacokinetics of a single oral dose of gabapentin suspension (11 mg/kg) in great horned owls ( Bubo virginianus ). Plasma gabapentin concentrations were determined in six healthy birds for 48 hr using high-performance liquid chromatography with mass spectrometric detection. Plasma gabapentin concentrations were estimated by noncompartmental pharmacokinetic analysis. The harmonic mean (±SD) maximum concentration (Cmax), time to maximum concentration (Tmax), and elimination half-life (tv2λZ) for gabapentin (11 mg/kg) were 6.17±0.83 µg/ml, 51.43±5.66 min, and 264.60±69.35 min, respectively. In this study, plasma gabapentin concentrations were maintained above 2 µg/ml for 528 min (8.8 hr), suggesting that gabapentin administered orally every 8 hr may be appropriate in great horned owls.

Oxidative stress biomarkers in Eurasian eagle owls (Bubo bubo) in three different scenarios of heavy metal exposure.

The main aim of the present study is the assessment of oxidative stress related to metals in the Eurasian eagle owl (Bubo bubo) from three areas (agricultural and rural area, industrial area, and mining area) of Murcia, Southern Spain. Mean blood metal concentrations were Cd=0.07±0.21, Pb=3.27±5.21, Cu=10.62±4.77, Zn=311.47±67.14, Hg=2.32±3.83 µg/dl wet weight. Although individuals from the mining area had significant higher Pb and Hg concentrations, and significant lower glutathione-S-transferase (GST) and catalase (CAT) activities in red blood cells (RBC); the lack of differences in oxidative damage to membrane lipids (TBARS) among areas suggests that the antioxidant capacity of the different populations is able to deal with oxidant species and maintain TBARS levels in the same amount. Despite the low levels of metals, several oxidative stress biomarkers were correlated with metal concentrations. This study provides threshold concentrations at which metals cause effects on the antioxidant system in Eagle owls. Blood Cd concentrations greater than 0.3 µg/dl produced an inhibition in GPx (32%) and CAT (26%) activity in RBC. However, Cd concentrations higher than 0.02 µg/dl were enough to produce an inhibition of these enzymes. Regarding Pb levels, blood concentrations above 2 µg/dl produced an inhibition of 8% and 10.5% in GPx and CAT activities, respectively, in RBC. A depletion of 16% and 4% in tGSH levels was associated with Pb concentrations higher than 15 and 3 µg/dl, respectively, in individuals from the ancient mine site. In addition, Pb concentrations above 2 and 10 µg/dl produced a TBARS induction of 10% and 28%, respectively, in individuals from both the industrial and the mining area. Finally, Hg concentrations greater than 3 and 10 µg/dl resulted in a TBARS induction of 102% and 190%, respectively, in Eurasian eagle owls from the industrial area. Our findings show that Pb may produce effects on oxidative stress biomarkers in Strigiformes at lower concentrations than those typically accepted for considering physiological effects in Falconiformes (20 µg/dl in blood). In addition, we provide new data on Hg and Cd concentrations related to effects in the antioxidant system.

Hematologic and plasma biochemistry reference intervals of healthy adult barn owls (Tyto alba).

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.

Pharmacokinetics of a single dose of intravenous and oral meloxicam in red-tailed hawks (Buteo jamaicensis) and great horned owls (Bubo virginianus).

Pharmacokinetic data were determined after a single dose of meloxicam in red-tailed hawks (RTH; Buteo jamaicensis) and great horned owls (GHO; Bubo virginianus). In a nonrandomized crossover design, individual birds of each species received 1 dose of intravenous meloxicam (0.5 mg/kg i.v.; n = 7 for each species) followed by a 2-week washout period, and then each received 1 dose of oral meloxicam (0.5 mg/kg PO; n = 5 for each species). Blood samples were collected intermittently after administration, and meloxicam was detected in plasma by high-performance liquid chromatography. Time versus plasma concentration data were subjected to noncompartmental analysis. Red-tailed hawks were determined to have the shortest elimination half-life for meloxicam (0.49 +/- 0.5 hours) of any species documented. Great horned owls also eliminated meloxicam very rapidly (0.78 +/- 0.52 hours). Great horned owls achieved higher plasma concentrations (368 +/- 87 ng/mL) of meloxicam than RTH (182 +/- 167 ng/mL) after oral administration, although RTH had a markedly higher volume of distribution (832 +/- 711 mL/kg) than GHO (137.6 +/- 62.7 mL/kg). The differences in meloxicam pharmacokinetics between these 2 raptor species supports the need for species-dependent studies and underlines the challenges of extrapolating drug dosages between species. Results of this study suggest that the current recommended once-daily dosing interval of oral meloxicam is unlikely to maintain plasma concentrations anticipated to be therapeutic in either RTH or GHO, and practical dosing options are questionable for this nonsteriodal anti-inflammatory drug in these raptor species.

Pale and dark reddish melanic tawny owls differentially regulate the level of blood circulating POMC prohormone in relation to environmental conditions.

Knowledge of the hormonal pathway controlling genotype-specific norms of reaction would shed light on the ecological factors to which each genotype is adapted. Environmentally mediated changes in the sign and magnitude of covariations between heritable melanin-based colouration and fitness components are frequent, revealing that extreme melanin-based phenotypes can display different physiological states depending on the environment. Yet, the hormonal mechanism underlying this phenomenon is poorly understood. One novel hypothesis proposes that these covariations stem from pleiotropic effects of the melanocortin system. Melanocortins are post-translationally modified bioactive peptides derived from the POMC prohormone that are involved in melanogenesis, anti-inflammation, energy homeostasis and stress responses. Thus, differential regulation of fitness components in relation to environmental factors by pale and dark melanic individuals may be due to colour-specific regulation of the POMC prohormone. Accordingly, we found that the degree of reddish melanic colouration was negatively correlated with blood circulating levels of the POMC prohormone in female tawny owls (Strix aluco) rearing a brood for which the size was experimentally reduced, but not when enlarged, and in females located in rich but not in poor territories. Our findings support the hypothesis that the widespread links between melanin-based colouration and fitness components may be mediated, at least in part, by the melanocortin system.

Blood lead levels and δ-ALAD inhibition in nestlings of Eurasian Eagle Owl (Bubo bubo) to assess lead exposure associated to an abandoned mining area.

In order to biomonitor lead contamination in Southeastern Spain, 218 blood samples from 28 to 30-day old Eurasian Eagle Owl chicks (Bubo bubo) born between 2003 and 2007 were analysed. In general, mean lead levels showed that chicks were exposed to background concentrations. However, mean levels in chicks born in an ancient and abandoned mining site ("Sierra Minera Cartagena-La Union") or in their surroundings (Geometric mean (GM) = 5.83 µg/dl, range 0.49-25.61 µg/dl), an area highly polluted by lead and other metals, were significantly higher (p < 0.001) than the rest of the population (GM = 1.66 µg/dl, range = Non detected-18.37 µg/dl). Because δ-ALAD activity is considered the best biomarker for lead exposure and effect in birds, the activity of this enzyme was also evaluated and correlated with lead levels in blood. In this study, low levels of blood lead inhibited δ-ALAD, even when lead concentrations were lower than the limits described by other authors in raptors. Adverse effects caused by this inhibition may occur when blood lead levels were above 15 µg/dl, although only eight chicks presented these concentrations in their blood. Sampling site also influenced enzymatic activity, since it decreased about 60% in the polluted area in relation to the rest. For all these reasons, further research regarding risk assessment for lead exposure in Eagle Owls nesting in the polluted area is advisable. Our results suggest that the Eurasian Eagle Owl can be considered a suitable sentinel animal for monitoring lead contamination and δ-ALAD activity can be used as a sensitive biomarker for lead exposure and effect in this species.

Account and Utilization of Blood Lipid Profiles: Lipid Levels Predicted Hemosporidian Infection in Migrating Northern Saw-Whet Owls of Eastern North America.

Noting lipidomic changes following the parasitism of migrating birds, the metabolic needs of which are primarily fueled by lipids, can deepen our understanding of host-parasite interactions. We identified lipids of migrating Northern saw-whet owls (Aegolius acadicus) using collision-induced dissociation mass spectrometry, compared the lipidomic signatures of hemoparasite-infected and noninfected individuals, and performed cross-validation analyses to reveal associations between parasite infection and lipid levels. We found significantly lower levels of lipid classes phosphatidic acid (PA), phosphatidylglycerol (PG), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidylethanolamine (PE), phosphatidylcholine (PC), and sphingomyelin (SM) in infected Northern saw-whet owls than in the noninfected individuals. Conversely, we found higher levels for certain lysoPS and lysoPE species, and variable lipid level changes for free fatty acid (FFA) species. Reporting lipidomic changes observed between hemosporidian-infected and noninfected Northern saw-whet owls can strengthen our understanding of the mechanisms governing parasite proliferation in this species. Furthermore, our analysis indicated that lipidomic signatures are better predictors of parasite infection than the log-adjusted mass/wing chord body index, a metric commonly used to assess the influence of hemosporidia infection on the health of birds. Establishing a lipidomic profile for Northern saw-whet owls that provides baseline lipid levels during fall migration may assist future studies assessing causes of reductions in breeding brought about from subtle differences in behaviors such as delayed migration.

Temporal variation in glucocorticoid levels during the resting phase is associated in opposite way with maternal and paternal melanic coloration.

Sex-dependent selection can help maintain sexual dimorphism. When the magnitude of selection exerted on a heritable sex trait differs between the sexes, it may prevent each sex to reach its phenotypic optimum. As a consequence, the benefit of expressing a sex trait to a given value may differ between males and females favouring sex-specific adaptations associated with different values of a sex trait. The level of metabolites regulated by genes that are under sex-dependent selection may therefore covary with the degree of ornamentation differently in the two sexes. We investigated this prediction in the barn owl, a species in which females display on average larger black spots on the plumage than males, a heritable ornament. This melanin-based colour trait is strongly selected in females and weakly counter-selected in males indicating sex-dependent selection. In nestling barn owls, we found that daily variation in baseline corticosterone levels, a key hormone that mediates life history trade-offs, covaries with spot diameter displayed by their biological parents. When their mother displayed larger spots, nestlings had lower corticosterone levels in the morning and higher levels in the evening, whereas the opposite pattern was found with the size of paternal spots. Our study suggests a link between daily regulation of glucocorticoids and sex-dependent selection exerted on sexually dimorphic melanin-based ornaments.

Regulation of free corticosterone and CBG capacity under different environmental conditions in altricial nestlings.

The concentration of circulating glucocorticoids is regulated in response to environmental and endogenous conditions. Total circulating corticosterone, the main glucocorticoid in birds, consists of a fraction which is bound to corticosterone-binding globulins (CBG) and a free fraction. There is increasing evidence that the environment modulates free corticosterone levels through varying the concentration of CBG, but experimental evidence is lacking. To test the hypothesis that the regulation of chronic stress in response to endogenous and environmental conditions involves variation in both corticosterone release and CBG capacity, we performed an experiment with barn owl (Tyto alba) nestlings in two different years with pronounced differences in environmental conditions and in nestlings experimentally fed ad libitum. In half of the individuals we implanted a corticosterone-releasing pellet to artificially increase corticosterone levels and in the other half we implanted a placebo pellet. We then repeatedly collected blood samples to measure the change in total and free corticosterone levels as well as CBG capacity. The increase in circulating total corticosterone after artificial corticosterone administration varied with environmental conditions and with the food regime of the nestlings. The highest total corticosterone levels were found in nestlings growing up in poor environmental conditions and the lowest in ad libitum fed nestlings. CBG was highest in the year with poor environmental conditions, so that, contrary to total corticosterone, free corticosterone levels were low under poor environmental conditions. When nestlings were fed ad libitum total corticosterone, CBG and free corticosterone did not increase when administering corticosterone. These results suggest that depending on the individual history an animal experienced during development the HPA-axis is regulated differently.

Changes in Detected Anticoagulant Rodenticide Exposure in Barn Owls ( Tyto alba) in Kentucky, USA, in 2012-16.

Anticoagulant rodenticides (ARs) are widely used across North America to control rodent infestations but may cause direct mortality or nonlethal effects when secondarily consumed by raptors. Barn Owls ( Tyto alba) are at high risk for secondary consumption because they specialize in rodent prey and often live in human-made structures. We investigated the exposure of Barn Owls in Kentucky, US, to ARs and to dicoumarol, an anticoagulant compound naturally found in certain moldy forages. We tested the liver tissue of 48 Barn Owl carcasses collected during 2012-16. We confirmed exposure to one or more ARs in 33% of the birds examined and detected dicoumarol in 13% of the samples. Rodenticides detected included brodifacoum, coumachlor, and bromadiolone. The prevalence of detected exposure to brodifacoum for after-hatch-year birds (65%) was significantly ( P=0.012) higher than hatch-year birds (22%). Brodifacoum was the most commonly detected AR, found in 88% of AR-positive birds. The pesticide registration for this chemical in the US was canceled in 2015 for general consumer products, which likely resulted in a decreasing rate of detected exposure to brodifacoum during our study. We present these results as an example of secondary exposure rates during a period when a pesticide has been restricted and then removed from the consumer market.

Haemosporidian blood parasites in European birds of prey and owls.

Avian blood parasites have been intensively studied using morphological methods with limited information on their host specificity and species taxonomic status. Now the analysis of gene sequences, especially the mitochondrial cytochrome b gene of the avian haemosporidian species of Haemoproteus, Plasmodium, and Leucocytozoon, offers a new tool to review the parasite specificity and status. By comparing morphological and genetic techniques, we observed nearly the same overall prevalence of haemosporidian parasites by microscopy (19.8%) and polymerase chain reaction (PCR) (21.8%) analyses. However, in contrast to the single valid Leucocytozoon species (L. toddi) in the Falconiformes we detected 4 clearly distinctive strains by PCR screening. In the Strigiformes, where the only valid Leucocytozoon species is L. danilewskyi, we detected 3 genetically different strains of Leucocytozoon spp. Two strains of Haemoproteus spp. were detected in the birds of prey and owls examined, whereas the strain found in the tawny owl belonged to the morphospecies Haemoproteus noctuae. Three Plasmodium spp. strains that had already been found in Passeriformes were also detected in the birds of prey and owls examined here, supporting previous findings indicating a broad and nonspecific host spectrum bridging different bird orders.

Pharmacokinetics of butorphanol tartrate in red-tailed hawks (Buteo jamaicensis) and great horned owls (Bubo virginianus).

OBJECTIVE: To determine the pharmacokinetics of butorphanol tartrate after IV and IM single-dose administration in red-tailed hawks (RTHs) and great horned owls (GHOs). ANIMALS: 6 adult RTHs and 6 adult GHOs. PROCEDURES: Each bird received an injection of butorphanol (0.5 mg/kg) into either the right jugular vein (IVj) or the pectoral muscles in a crossover study (1-week interval between treatments). The GHOs also later received butorphanol (0.5 mg/kg) via injection into a medial metatarsal vein (IVm). During each 24-hour postinjection period, blood samples were collected from each bird; plasma butorphanol concentrations were determined via liquid chromatography-mass spectrometry. RESULTS: 2- and 1-compartment models best fit the IV and IM pharmacokinetic data, respectively, in both species. Terminal half-lives of butorphanol were 0.94 +/- 0.30 hours (IVj) and 0.94 +/- 0.26 hours (IM) for RTHs and 1.79 +/- 1.36 hours (IVj), 1.84 +/- 1.56 hours (IM), and 1.19 +/- 0.34 hours (IVm) for GHOs. In GHOs, area under the curve (0 to infinity) for butorphanol after IVj or IM administration exceeded values in RTHs; GHO values after IM and IVm administration were less than those after IVj administration. Plasma butorphanol clearance was significantly more rapid in the RTHs. Bioavailability of butorphanol administered IM was 97.6 +/- 33.2% (RTHs) and 88.8 +/- 4.8% (GHOs). CONCLUSIONS AND CLINICAL RELEVANCE: In RTHs and GHOs, butorphanol was rapidly absorbed and distributed via all routes of administration; the drug's rapid terminal half-life indicated that published dosing intervals for birds may be inadequate in RTHs and GHOs.

Plasma biochemistry RIs and age effect in European Strigiformes.

BACKGROUND: Blood biochemistry and hematology are essential in the laboratory diagnosis of disease. In Strigiformes, little information regarding reference values and influence of different preanalytic factors is available, while age is known to have an effect on some biochemistry analytes, especially in early life characterized by a rapid growth rate and increase in body mass. OBJECTIVE: The objective of this study was to determine baseline data for 29 blood biochemistry variables in 5 species of Iberian Strigiformes assigned to different age classes. METHODS: Healthy nocturnal birds living in wildlife health centers of Catalonia, Northeastern Spain were assigned to different age classes (chicks, juveniles, adults where possible and available) and their blood was collected and analyzed for different standard biochemistry variables. Species included Tawny owls, Little owls, Long-eared owls, Scop owls, and Barn owls. RESULTS: A total of 276 clinically healthy animals were sampled, including between 179 chicks, 52 juveniles, and 45 adults. The RIs of cholesterol, albumin, and osmolality were the only variables that did not show interspecific variability. Common trends between age class groups were observed in 4 species. Phosphorus and calcium concentrations and ALP activities decreased with increasing age, while concentrations of sodium and γ-globulins tended to increase in the Strigidae family. Differences were concentrated in chicks and no differences were observed between juveniles and adults in any variable. CONCLUSIONS: Our results demonstrate the importance of defining biochemical RIs specific for certain age classes.

Validation of a QuECheRS method for analysis of neonicotinoids in small volumes of blood and assessment of exposure in Eurasian eagle owl (Bubo bubo) nestlings.

Neonicotinoid pesticides have gained great interest in the last years both for agricultural and domestic use. Since the information on their environmental distribution or the effects derived from exposure to ecosystems and biota is scarce, new analytical techniques are being developed for monitoring studies. In this sense, two extraction techniques based on QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) methodology to detect the neonicotinoids authorised in Spain (acetamiprid, clothianidin, dinotefuran, imidacloprid, thiacloprid, nitenpyram and thiamethoxam) were adapted and compared: a) using acetate buffer (AB); and b) using citrate buffer (CB). For detection and quantification, high performance liquid chromatography coupled with time of flight mass spectrometry (HPLC/TOF-MS) was used. The CB method provided a wide range of recoveries (68-134%) and accuracy (4-9%). The AB method provided good recoveries (59-76%, 59% corresponded to clothianidin) precision (4-11%) linearity (0.987-0.998%) and limit of quantification (2-10ng/mL) for all the compounds. To test the effectiveness of the technique, we analysed 30 blood samples of free-ranging nestlings of Eurasian eagle owl (Bubo bubo). The only compound detected, in one nestling from a dry land farming area, was imidacloprid, with a concentration of 3.28ng/mL. To our knowledge, this is the first study of neonicotinoid pesticides in free-ranging birds of prey using non-destructive samples, providing the first insight for biomonitoring studies. Further studies, including toxicokinetics and toxicodynamics, are recommended to assess the risk for these species.

Hematologic reference intervals and age effect in European Strigiformes.

BACKGROUND: The clinical importance of hematologic testing in avian veterinary medicine is reflected in the increasing number of studies for the establishment of hematologic RIs of Strigiformes and other species. Age is an important physiologic factor in birds and the effect on hematology variable should be understood. OBJECTIVES: The objective of this study was to determine baseline data of hematologic variables in 5 species of Iberian Strigiformes in different age classes. METHODS: Nocturnal birds of prey were sampled at Wildlife Health Centers. Packed cell volume was determined by the microhematocrit centrifugation method, and RBC and WBC counts were determined using the direct hemocytometer count method with Natt and Herrick solution. Hemoglobin concentration was measured spectrophotometrically. The MCV, MCHC, and MHC were calculated using the standard formulas. The differential WBC count was performed by the routine microscopic evaluation of 200 cells on a blood smear manually stained with Wright stain. Thrombocyte blood count estimate was obtained from the blood film. RESULTS: No differences were observed between juveniles and adults for any variable evaluated in Tawny owl, Little owl, Scops owl, Long-eared owl, and Barn owl. In addition, PCV, RBC, and HGB of chicks were statistically significantly lower than in juveniles and adults, and total WBC was significantly higher in Tawny owl, Little owl, Scops owl, and Long-eared owl. CONCLUSIONS: Our findings provide evidence that laboratory data from chicks of Strigiformes are different compared to juveniles and adults; therefore, separate RIs were defined.