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Chronic arsenic (As) exposure is a major environmental threat to human health affecting >100 million people worldwide. Low blood selenium (Se) increases the risk of As-induced health problems. Our aim was to reduce As toxicity through a naturally Se-rich lentil diet. In a randomized, double-blind, placebo-control trial in Bangladesh, 405 participants chronically exposed to As were enrolled. The intervention arm (Se-group) consumed Se-rich lentils (55 μg Se/day); the control arm received lentils of similar nutrient profile except with low Se (1.5 μg Se/day). Anthropometric measurements, blood, urine and stool samples, were taken at baseline, 3 and 6 months; hair at baseline and 6 months after intervention. Morbidity data were collected fortnightly. Measurements included total As in all biological samples, As metabolites in urine, and total Se in blood and urine. Intervention with Se-rich lentils resulted in higher urinary As excretion (p = 0.001); increased body mass index (p ≤ 0.01), and lower incidence of asthma (p = 0.05) and allergy (p = 0.02) compared to the control group. The Se-group demonstrated increased excretion of urinary As metabolite, dimethylarsinic acid (DMA) at 6 months compared to control group (p = 0.008). Consuming Se-rich lentils can increase As excretion and improve the health indicators in the presence of continued As exposure., (Copyright © 2019. Published by Elsevier Inc.)

Background: In patients with obesity-related hypertension (ORH), reaction to antihypertensive medication is likely influenced by patientcharacteristics., Methods: Effects of aliskiren, moxonidine and hydrochlorothiazide on 24-h blood pressure (BP) were compared to placebo. Linear mixed effect models were used to analyze the effect of patient characteristics on BP levels and treatment response., Results: Systolic BP response to aliskiren was higher in patients with a BMI > 30.7 kg/m2 compared to patients with a BMI ≤ 30.7 kg/m2 (-21 mmHg versus -4 mmHg). In patients with a hsCRP > 1.8 mg/L the systolic BP response to aliskiren was higher than in patients with a low hsCRP (-15 mmHg versus -7 mmHg). Hydrochlorothiazide (HCTZ) treatment effect on systolic BP was -13 mmHg when heart rate > 71 beats/min compared to -3 mmHg when heart rate was ≤ 71 beats/min., Conclusion: In patients with ORH, BP response to aliskiren is positively related to BMI and hsCRP. Systolic BP response to HCTZ is positively related to heart rate and negatively to renin levels., Trial Registration: NCT01138423. Registered June 4th, 2010.

Numerous studies conducted to assess welfare of domestic dogs housed in kennel facilities have reported that these dogs experience suboptimal living conditions. One important goal of improving welfare of kennelled dogs is to reduce their stress levels, and one recommended approach for improving welfare of kennelled dogs is group or social housing. The beneficial effects of management changes designed to achieve this goal should be measurable in individual animals. Stress is evident through behaviours exhibited, as well as via the concentration of cortisol, a key hormone reflecting stress. Using behavioural and hair cortisol measures, we conducted a pilot study to measure the impact of switching dogs housed in a long-term kennels facility from solitary to pair housing, using both within-subjects and between-groups comparisons. Considerable individual variation in dog responses was noted, with only two of eight pair-housed dogs showing significant declines in multiple stress-related behaviours once in pair housing. The most sensitive behaviours were active vigilance and repetitive movements (such as jumping and pacing). Barking was reduced overall in the facility following the housing change, even among dogs still in solitary housing. The long-term stress as reflected in hormone deposition in hair also provided encouraging indications that the dogs experienced lower stress levels when in paired housing; dogs showed a significant decline in hair cortisol levels from the first (prehousing change) to second (postintervention) samples. Domestic dogs are social animals, and numerous indications of potential benefit were recorded with no negative impacts seen. Based on our findings, we recommend pair or group housing of compatible dogs as a promising addition to the strategies available to those seeking to improve welfare of kennelled dogs. Future studies using higher numbers of animals and that include tracking of hair cortisol, vigilance behaviour, repetitive movements and barking would be desirable., Competing Interests: Competing interests: None declared.

Background: Millions of people worldwide are exposed to dangerous levels of arsenic (above the WHO water standard of 10 ppb) in drinking water and food. Lack of nutritious foods exacerbates the adverse health effects of arsenic poisoning. The micronutrient selenium is a known antagonist to arsenic, promoting the excretion of arsenic from the body. Studies are in progress examining the potential of using selenium supplement pills to counteract arsenic toxicity. We are planning a clinical trial to test whether high-selenium lentils, as a whole food solution, can improve the health of arsenic-exposed Bangladeshi villagers., Methods/design: A total of 400 participants (about 80 families) will be divided into two groups via computer-generated block randomization. Eligibility criteria are age (≥14) years) and arsenic concentration in the household tube well (≥100 ppb). In this double-blind study, one group will eat high-selenium lentils grown in western Canada; the other will consume low-selenium lentils grown in Idaho, USA. Each participant will consume 65 g of lentils each day for 6 months. At the onset, midterm, and end of the trial, blood, urine and stool, plus hair (day 1 and at 6 months only) samples will be collected and a health examination conducted including assessment of acute lung inflammation, body mass and height, and blood pressure. The major outcome will be arsenic excretion in urine and feces, as well as arsenic deposition in hair and morbidity outcomes as assessed by a biweekly questionnaire. Secondary outcomes include antioxidant status, lipid profile, lung inflammation status, and blood pressure., Discussion: Selenium pills as a treatment for arsenic exposure are costly and inconvenient, whereas a whole food approach to lower the toxic burden of arsenic may be a practical remedy for Bangladeshi people while efforts to provide safe drinking water are continuing. If high-selenium lentils prove to be effective in counteracting arsenic toxicity, agronomic partnerships between Canada and Bangladesh will work to improve the selenium content of the Bangladeshi-grown lentil crops. Results will be presented to the community to promote informed food choices, which may include increasing selenium in their diet., Trial Registration: ClinicalTrials.gov NCT02429921.

Background: Cardiovascular disease (CVD) is a major cause of death worldwide, and arsenic (As) intake, mainly through drinking water, is a well-known risk factor for CVD as well as other health problems. Selenium (Se) is a known antagonist to As toxicity., Objective: We tested the potential of high-Se lentils from the Canadian prairies as a therapeutic food to alter the outcome of As-enhanced atherosclerosis., Materials and Methods: Male ApoE(-/-) mice exposed to a moderate level of As (200ppb) in their drinking water, and control mice on tap water received one of three lentil diets: Se-deficient (0.009mg/kg), Se-adequate (0.16mg/kg) or Se-high (0.3mg/kg). After 13weeks, lesion formation in the aortic arch and sinus were assessed. Intralesional cellular composition, serum lipid levels and hepatic oxidative stress were assessed as well., Results: Arsenic-exacerbated plaque formation was reduced in the sinus and completely abolished in the aortic arch of mice on the Se-fortified lentil diet, whereas lesions were increased in As-exposed mice on both the Se-deficient and Se-adequate diets. Notably, Se deficiency contributed to proatherogenic composition of serum lipids in As-exposed mice as indicated by high-density lipoprotein:low-density lipoprotein. At least adequate Se status was crucial for counteracting As-induced oxidative stress., Conclusion: This study is the first to show the potential of high-Se lentils to protect against As-triggered atherosclerosis, and this invites further investigations in human populations at risk from As contamination of their drinking water., (Crown Copyright © 2015. Published by Elsevier Inc. All rights reserved.)

Increasing activity of oil sands extraction and processing in northern Alberta is marked by ongoing controversy about the nature and extent of associated environmental impacts. Bitumen contains a mixture of toxic chemicals, including metals and residual polycyclic aromatic hydrocarbons (PAHs), whose release into the environment poses a distinct risk to the surrounding environment, plus wildlife and human health. In the present study, the authors evaluated several subclinical biomarkers of exposure and effect to mixtures of metals (Pb, Cd, and Hg) and/or PAHs (3 alkylated forms) at environmentally relevant concentrations (100-fold and 10-fold higher than the maximum dissolved concentrations found in snow, to simulate a worst-case scenario), using laboratory mice as a model for future studies of small mammals in the wild. Both metals and alkyl-PAHs exposure were associated with 1) increased relative liver, kidney, and spleen size; 2) alterations in the homeostasis of the antioxidant vitamins A and E in liver; and 3) compromised glutathione redox status in testes, with results also indicating synergistic interactions from co-exposure. The combination of morphometric and oxidative stress biomarkers provide reliable and sensitive measures of the response to contaminant exposure in a mammalian model, suggesting associated physiological costs. Based on the present experimental study, the authors propose that wild small mammals will prove to be valuable sentinel species reflecting sublethal health effects from oil sands-related contaminants. The present study's results also present a basis for the interpretation of future field data., (© 2015 SETAC.)

Exposure to water containing petroleum waste products can generate both overt and subtle toxicological responses in wildlife, including birds. Such exposure can occur in the tailings ponds of the mineable oil sands, which are located in Alberta, Canada, under a major continental flyway for waterfowl. Over the 40 year history of the industry, a few thousand bird deaths have been reported following contact with bitumen on the ponds, but a new monitoring programme demonstrated that many thousands of birds land annually without apparent harm. This new insight creates an urgent need for more information on the sublethal effects on birds from non-bitumen toxicants that occur in the water, including naphthenic acids, polycyclic aromatic hydrocarbons, heavy metals and salts. Ten studies have addressed the effects of oil sands process-affected water (OSPW), and none reported acute or substantial adverse health effects. Interpretive caution is warranted, however, because nine of the studies addressed reclaimed wetlands that received OSPW, not OSPW ponds per se, and differences between experimental and reference sites may have been reduced by shared sources of pollution in the surrounding air and water. Two studies examined eggs of birds nesting >100 km from the mine sites. Only one study exposed birds directly and repeatedly to OSPW and found no consistent differences between treated and control birds in blood-based health metrics. If it is true that aged forms of OSPW do not markedly affect the health of birds that land briefly on the ponds, then the extensiveness of current bird-deterrent programmes is unwarranted and could exert negative net environmental effects. More directed research on bird health is urgently needed, partly because birds that land on these ponds subsequently migrate to destinations throughout North America where they are consumed by both humans and wildlife predators.

Epidemiological studies have shown that arsenic exposure increases atherosclerosis, but the mechanisms underlying this relationship are unknown. Monocytes, macrophages and platelets play an important role in the initiation of atherosclerosis. Circulating monocytes and macrophages bind to the activated vascular endothelium and migrate into the sub-endothelium, where they become lipid-laden foam cells. This process can be facilitated by platelets, which favour monocyte recruitment to the lesion. Thus, we assessed the effects of low-to-moderate arsenic exposure on monocyte adhesion to endothelial cells, platelet activation and platelet-monocyte interactions. We observed that arsenic induces human monocyte adhesion to endothelial cells in vitro. These findings were confirmed ex vivo using a murine organ culture system at concentrations as low as 10 ppb. We found that both cell types need to be exposed to arsenic to maximize monocyte adhesion to the endothelium. This adhesion process is specific to monocyte/endothelium interactions. Hence, no effect of arsenic on platelet activation or platelet/leukocyte interaction was observed. We found that arsenic increases adhesion of mononuclear cells via increased CD29 binding to VCAM-1, an adhesion molecule found on activated endothelial cells. Similar results were observed in vivo, where arsenic-exposed mice exhibit increased VCAM-1 expression on endothelial cells and increased CD29 on circulating monocytes. Interestingly, expression of adhesion molecules and increased binding can be inhibited by antioxidants in vitro and in vivo. Together, these data suggest that arsenic might enhance atherosclerosis by increasing monocyte adhesion to endothelial cells, a process that is inhibited by antioxidants.

In the oil sands of Alberta, Canada, toxicology research has largely neglected the effects of air contaminants on biota. Captive Japanese quail (Coturnix c. japonica) and American kestrels (Falco sparverius) were exposed to mixtures of volatile organic compounds and oxidizing agents (benzene, toluene, NO2 and SO2) in a whole-body inhalation chamber, to test for toxicological responses. Hepatic biotransformation measured through 7-ethoxyresorufin-O-dealkylase (EROD) tended to be increased in exposed kestrels (p=0.06) but not in quail (p=0.15). Plasma corticosterone was increased in the low dose group for quail on the final day of exposure (p=0.0001), and midway through the exposure period in exposed kestrels (p=0.04). For both species, there was no alteration of T and B-cell responses, immune organ mass, or histology of immune organs (p>0.05). This study provides baseline information valuable to complement toxicology studies and provides a better understanding of potential health effects on wild avifauna., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Changes in environmental and wildlife health from contaminants in tailings water on the Canadian oil sands have been well-studied; however, effects of air contaminants on wildlife health have not. A field study was conducted to assess biological costs of natural exposure to oil sands-related air emissions on birds. Nest boxes for tree swallows (Tachycineta bicolor) were erected at two sites; within 5 km of active oil sands mining and extraction, and ≥ 60 km south, at one reference site. Passive air monitors were deployed at the nest boxes to measure nitrogen dioxide, sulfur dioxide, ozone, volatile organic compounds, and polycyclic aromatic hydrocarbons (PAHs). Nestlings were examined at day 9 post hatching to assess T cell function and morphometry. At day 14 post hatching, a subset of nestlings was euthanized to measure detoxification enzymes, endocrine changes, and histological alterations of immune organs. Except for ozone, all air contaminants were higher at the two oil sands sites than the reference site (up to 5-fold). Adult birds had similar reproductive performance among sites (p>0.05). Nestlings from industrial sites showed higher hepatic ethoxyresorufin O-dealkylase (EROD) induction (p<0.0001) with lower relative hepatic mass (p=0.0001), a smaller T cell response to the phytohemagglutinin skin test (p=0.007), and smaller bursae of Fabricius (p<0.02); a low sample size for one site indicating lower body condition scores (p=0.01) at day 14 warrants cautious interpretation. There were no differences among nestlings for feather corticosterone (p>0.6), and no histological alterations in the spleen or bursa of Fabricius (p>0.05). This is the first report examining toxicological responses in wild birds exposed to air contaminants from industrial activity in the oil sands. It is also the first time that small, individual air contaminant monitors have been used to determine local contaminant levels in ambient air around nest boxes of wild birds., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Food availability can influence the nutritional and social dynamics within and among species. Our investigation focused on grizzly and black bears in coastal British Columbia, Canada, where recent and dramatic declines in their primary prey (salmon) raise concerns about potentially negative effects on bear physiology. We examined how salmon availability relates to stress and reproductive hormones in coastal grizzly (n = 69) and black bears (n = 68) using cortisol and testosterone. In hair samples from genotyped individuals, we quantified salmon consumption using stable isotope analysis and hormone levels by enzyme immunoassay. To estimate the salmon biomass available to each bear, we developed a spatially explicit approach based on typical bear home-range sizes. Next, we compared the relative importance of salmon consumption and salmon availability on hormone levels in male bears using an information theoretical approach. Cortisol in grizzly bears was higher in individuals that consumed less salmon, possibly reflecting nutritional stress. In black bears, cortisol was better predicted by salmon availability than salmon consumption; specifically, individuals in areas and years with low salmon availability showed higher cortisol levels. This indicates that cortisol in black bears is more strongly influenced by the socially competitive environment mediated by salmon availability than by nutritional requirements. In both species, testosterone generally decreased with increasing salmon availability, possibly reflecting a less competitive environment when salmon were abundant. Differences between species could relate to different nutritional requirements, social densities and competitive behaviour and/or habitat use. We present a conceptual model to inform further investigations in this and other systems. Our approach, which combines data on multiple hormones with dietary and spatial information corresponding to the year of hair growth, provides a promising tool for evaluating the responses of a broad spectrum of wildlife to changes in food availability or other environmental conditions.

Bitumen extraction from the oil sands of northern Alberta produces large volumes of process-affected water that contains substances toxic to wildlife. Recent monitoring has shown that tens of thousands of birds land on ponds containing this water annually, creating an urgent need to understand its effects on bird health. We emulated the repeated, short-term exposures that migrating water birds are thought to experience by exposing pekin ducks (Anas platyrhynchos domestica) to recycled oil sands process-affected water (OSPW). As indicators of health, we measured a series of physiological (electrolytes, metabolites, enzymes, hormones, and blood cells) and toxicological (metals and minerals) variables. Relative to controls, juvenile birds exposed to OSPW had higher potassium following the final exposure, and males had a higher thyroid hormone ratio (T3/T4). In adults, exposed birds had higher vanadium, and, following the final exposure, higher bicarbonate. Exposed females had higher bile acid, globulin, and molybdenum levels, and males, higher corticosterone. However, with the exception of the metals, none of these measures varied from available reference ranges for ducks, suggesting OSPW is not toxic to juvenile or adult birds after three and six weekly, 1 h exposures, but more studies are needed to know the generality of this result.

Tree swallows (Tachycineta bicolor) inhabiting reclaimed wetlands on the oil sands in northern Alberta are potentially exposed to elevated levels of oil sands constituents such as polycyclic aromatic compounds (PAC) through diet. While increased detoxification enzyme activity as measured using 7-ethoxyresorufin O-deethylase in nestlings is a generally accepted indicator of exposure to oil sands constituents, there is no apparent method to detect dietary exposure specific to oil sands processed material (OSPM). In this study, stable C and N isotopes were analyzed from muscle and feathers of nestling tree swallows (15 d old) to distinguish dietary exposure of birds near reference and OSPM wetlands. High δ¹⁵N and low δ¹³C values in the nestling tissues differentiated those from the OSPM wetlands and reference sites. Lower δ¹⁵N values of nestlings compared to the δ¹⁵N values of larval chironomids from an earlier study suggested that the majority of the diet of the nestlings was derived from non-OSPM sources, despite residence near and on the OSPM wetlands. Our finding of limited utilization of OSPM resources by tree swallows indicates either low abundance or diversity of dietary items emerging from OSPM wetlands, or sensory avoidance of prey from those wetlands. Minimal consumption of OSPM-derived dietary sources may be attributed to published findings of limited adverse effects on tree swallow reproduction, or growth and development for these same nestlings. This study demonstrated that stable isotope analysis, particularly for N isotopes, may serve as a useful tool to trace dietary exposure to OSPM constituents as part of avian ecotoxicology assessments of reclaimed wetlands on the oil sands.

Background: The association between depression and metabolic syndrome is becoming more obvious. Waist circumference (WC) might be the most important metabolic syndrome (MetS) feature in relation to late-life depression, with a possible mediating role for adiponectin., Methods: Cross-sectional population based survey of 1277 participants (50-70 years). We measured all components of MetS, plasma adiponectin levels and depressive symptoms using Beck Depression Inventory (BDI). Principal components analysis on the BDI items revealed two factors, representing a cognitive-affective and a somatic-affective symptom-cluster. Multiple linear regression models with the BDI sum score and both depression symptom-clusters as dependent variables, respectively, were used to examine the association with each component of metabolic syndrome adjusted for confounders. We explored sex-differences as well as a hypothesised mediating effect of adiponectin., Results: The presence of MetS as well as number of metabolic risk factors were significantly associated with BDI sum score. In men WC, triglycerides and HDL cholesterol explained variance in depressive symptoms, whereas in women this effect was confined to WC. Moreover, irrespective of sex, all associations were primarily driven by the somatic-affective symptom-cluster. Adiponectin neither mediated nor moderated any of the associations found., Limitations: Cross-sectional design limits causal interpretation. Being a population-based survey, some selection bias might have occurred toward healthier part of population., Conclusions: Although pathophysiological mechanisms underlying the association between metabolic disturbances and depression remains to be elucidated, our study points to sex-differences as well as a specific phenotype of depression that is associated with metabolic disturbances., (© 2013 Elsevier B.V. All rights reserved.)

Physiological indicators of social and nutritional stress can provide insight into the responses of species to changes in food availability. In coastal British Columbia, Canada, grizzly bears evolved with spawning salmon as an abundant but spatially and temporally constrained food source. Recent and dramatic declines in salmon might have negative consequences on bear health and ultimately fitness. To examine broadly the chronic endocrine effects of a salmon niche, we compared cortisol, progesterone, and testosterone levels in hair from salmon-eating bears from coastal BC (n = 75) with the levels in a reference population from interior BC lacking access to salmon (n = 42). As predicted, testosterone was higher in coastal bears of both sexes relative to interior bears, possibly reflecting higher social density on the coast mediated by salmon availability. We also investigated associations between the amount of salmon individual bears consumed (as measured by stable isotope analysis) and cortisol and testosterone in hair. Also as predicted, cortisol decreased with increasing dietary salmon and was higher after a year of low dietary salmon than after a year of high dietary salmon. These findings at two spatial scales suggest that coastal bears might experience nutritional or social stress in response to on-going salmon declines, providing novel insights into the effects of resource availability on fitness-related physiology.

This study evaluated the impact of oral perfluorooctanoic acid (PFOA) on Japanese quail at concentrations found in American and Belgian workers at PFOA manufacturing facilities. Three arms of the immune system were tested; T cell, B cell, and innate immunity. After 6 weeks exposure, quail were challenged with E. coli infection to test the ultimate measure of immunotoxicity, disease resistance. The T cell response was lower in the high exposure groups. Antibody mediated, and innate immune responses were not different. Growth rate was higher, whereas thyroid hormone levels were lower in PFOA-exposed birds. Morbidity/mortality from disease challenge was not different among the control and PFOA-exposed groups, and no overt PFOA toxicity was observed pre-disease challenge. Although PFOA at 'worst case scenario' levels caused T cell immunosuppression, this did not translate into increased disease susceptibility, demonstrating that immunotoxicity testing must be interpreted with caution since disease resistance is the ultimate concern., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Boreal wetlands play an important role in global carbon balance. However, their ecosystem function is threatened by direct anthropogenic disturbance and climate change. Oil sands surface mining in the boreal regions of Western Canada denudes tracts of land of organic materials, leaves large areas in need of reclamation, and generates considerable quantities of extraction process-affected materials. Knowledge and validation of reclamation techniques that lead to self-sustaining wetlands has lagged behind development of protocols for reclaiming terrestrial systems. It is important to know whether wetlands reclaimed with oil sands process materials can be restored to levels equivalent to their original ecosystem function. We approached this question by assessing carbon flows and food web structure in naturally formed and oil sands-affected wetlands constructed in 1970-2004 in the postmining landscape. We evaluated whether a prescribed reclamation strategy, involving organic matter amendment, accelerated reclaimed wetland development, leading to wetlands that were more similar to their natural marsh counterparts than wetlands that were not supplemented with organic matter. We measured compartment standing stocks for bacterioplankton, microbial biofilm, macrophytes, detritus, and zoobenthos; concentrations of dissolved organic carbon and residual naphthenic acids; and microbial production, gas fluxes, and aquatic-terrestrial exports (i.e., aquatic insect emergence). The total biomass of several biotic compartments differed significantly between oil sands and reference wetlands. Submerged macrophyte biomass, macroinvertebrate trophic diversity, and predator biomass and richness were lower in oil sands-affected wetlands than in reference wetlands. There was insufficient evidence to conclude that wetland age and wetland amendment with peat-mineral mix mitigate effects of oil sands waste materials on the fully aquatic biota. Although high variability was observed within most compartments, our data show that 20-year-old wetlands containing oil sands material have not yet reached the same level of function as their reference counterparts.

Birds have been widely used as sentinels of ecosystem health reflecting changes in habitat quality, increased incidence of disease, and exposure to and effects of chemical contaminants. Numerous studies addressing these issues focus on the breeding period, since hormonal, behavioural, reproductive, and developmental aspects of the health can be observed over a relatively short time-span. Many body systems within individuals are tightly integrated and interdependent, and can be affected by contaminant chemicals, disease, and habitat changes in complex ways. Animals higher in the food web will reflect cumulative effects of multiple stressors. Such features make birds ideal indicators for assessing environmental health in areas of environmental concern. Five case studies are presented, highlighting the use of different species which have provided insight into ecosystem sustainability, including (i) the consequences of anthropogenic disturbances of sagebrush habitat on the greater northern sage grouse Centrocercus urophasianus; (ii) the high prevalence of disease in very specific passerine species in the Canary Islands closely paralleling deterioration of formerly productive desert habitat and ensuing interspecific stressors; (iii) fractures, abnormal bone structure, and associated biochemical aberrations in nestling storks exposed to acidic tailings mud from a dyke rupture at an iron pyrite mine near Sevilla, Spain; (iv) newly presented data demonstrating biochemical changes in nestling peregrine falcons Falco peregrinus and associations with exposure to major chemical classes in the Great Lakes Basin of Canada; and (v) the variability in responses of tree swallows Tachycineta bicolor to contaminants, biological and meteorological challenges when breeding in the Athabasca oil sands., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Cortisol measurements of hair are becoming a valuable tool in monitoring chronic stress. To further validate this approach in domestic dogs, we compared the variability of cortisol immunoreactivity in hair with that in saliva and feces of dogs housed under constant social and physical conditions. Fecal (n = 268), and hair (n = 21) samples were collected over 3 mo from 7 dogs housed in a kennel and kept for training veterinary students in minimally invasive procedures. Salivary samples (n = 181) were collected 3 times daily twice weekly during the last month of the study. Hair and salivary samples were analyzed by enzyme immunoassay and feces by radioimmunoassay. HPLC coupled with tandem mass spectrometry was used to confirm the presence of cortisol in 3 hair samples. Variability of cortisol was compared across sample types by using repeated-measures ANOVA followed by paired t tests. Within dogs, cortisol immunoreactivity was less variable in hair than in saliva or feces. Averaged over time, the variability of fecal samples approached that of hair when feces were collected at least 4 times monthly. As predicted, the stable social and environmental condition of the dogs maintained repeatability over time and supported the hypothesis that data from hair samples reflect baseline cortisol levels. These findings indicate that determining cortisol immunoreactivity in hair is a more practical approach than is using samples of saliva or feces in monitoring the effects of long-term stressors such as social or physical environments and disease progression.

Acanthocephalans can be pathogenic helminths of marine birds. Every year during the breeding season, there is variable mortality among prefledged chicks from the largest known Olrog's gull (Larus atlanticus) colony. Mortality has been associated with infection by the acanthocephalan Profilicollis chasmagnathi. Our aim was to study the role of chicks' size as a risk factor for intensity of infection and severe pathology, and to expand upon previous pathological findings reported in acanthocephalan-infected chicks. Size of the chick was associated with intensity of infection and number of intestinal perforations, which increased by 6.9% and 4.1%, respectively, for each millimetre increment in chick size. Infection was associated with inflammatory enteritis and granulomatous peritonitis. Complete intestinal perforations were observed in 85% and 97.3% of the studied chicks in 2005 and 2006, respectively, and they were observed very early during the post-hatching period. Our results show: (1) the presence of advanced pathology associated with acanthocephalan infections in chicks, beginning very early in the post-hatching period; and (2) significant increases in the intensity of infection and the associated pathology as a function of size of chicks, in dead chicks during this period.

Trematodes and nematodes can be pathogenic helminths of birds. Every year during the breeding season, there is variable mortality among chicks from the largest Olrog's gull (Larus atlanticus) colony in Argentina. During two consecutive breeding seasons of Olrog's gull, we studied epidemiological and pathological aspects of infections by digeneans (Microphallidae) and nematodes (Acuariidae) in Olrog's gull chicks. Prevalence of nematode infection was 80.3% in 2005 and 89.2% in 2006, and mean intensity was 23.7 in 2005 and 50.8 in 2006. The risk for infection rose 34.3% and the intensity of infection 6.7% for every increase of 1 mm in head-beak length. The nematodes occupied the proventricular glands and caused disruption of their structure and mild inflammatory proventriculitis. Prevalence of digenean infection was 97.0% in 2005 and 97.3% in 2006. In 10-day-old live chicks, prevalence was 98.0% in 2006 and 95.3% in 2007. Infection was associated with severe catarrhal enteritis, lymphocyte/eosinophil-rich inflammatory responses, extensive fibroblast proliferation around the parasites, and disruption of the architecture of the adjacent crypts.

Naphthenic acids (NAs) have been cited as one of the main causes of the toxicity related to oil sands process-affected materials and have recently been measured in biological tissues (fish). However, adverse effects have not been a consistent finding in toxicology studies on vertebrates. This study set out to determine two factors: 1) whether exposure to commercial NAs (Refined Merichem) resulted in detectable tissue residues in native amphibians (northern leopard frogs, Lithobates pipiens), and 2) whether such exposure would produce clinical or subclinical toxicity. Frogs were kept in NA solutions (0, 20, or 40 mg/L) under saline conditions comparable to that on reclaimed wetlands in the Athabasca oil sands for 28 days. These exposures resulted in proportional NA concentrations in muscle tissue of the frogs, estimated by gas chromatography-mass spectrometry analyses. Detailed studies determined if the increasing concentrations of NAs, and subsequently increased tissue NA levels, caused a proportional compromise in the health of the experimental animals. Physiological investigations included innate immune function, thyroid hormone levels, and hepatic detoxification enzyme induction, none of which differed in response to increased exposures or tissue concentrations of NAs. Body mass did increase in both the salt- and NA-exposed animals, likely related to osmotic pressure and uptake of water through the skin. Our results demonstrate that commercial NAs are absorbed and deposited in muscle tissue, yet they show few negative physiological or toxicological effects on the frogs., (Copyright © 2012. Published by Elsevier B.V.)

Parasites are increasingly recognized for their profound influences on individual, population and ecosystem health. We provide the first report of gastrointestinal parasites in gray wolves from the central and north coasts of British Columbia, Canada. Across 60 000 km(2), wolf feces were collected from 34 packs in 2005-2008. At a smaller spatial scale (3300 km(2)), 8 packs were sampled in spring and autumn. Parasite eggs, larvae, and cysts were identified using standard flotation techniques and morphology. A subset of samples was analysed by PCR and sequencing to identify tapeworm eggs (n=9) and Giardia cysts (n=14). We detected ≥14 parasite taxa in 1558 fecal samples. Sarcocystis sporocysts occurred most frequently in feces (43·7%), followed by taeniid eggs (23·9%), Diphyllobothrium eggs (9·1%), Giardia cysts (6·8%), Toxocara canis eggs (2·1%), and Cryptosporidium oocysts (1·7%). Other parasites occurred in ≤1% of feces. Genetic analyses revealed Echinococcus canadensis strains G8 and G10, Taenia ovis krabbei, Diphyllobothrium nehonkaiense, and Giardia duodenalis assemblages A and B. Parasite prevalence differed between seasons and island/mainland sites. Patterns in parasite prevalence reflect seasonal and spatial resource use by wolves and wolf-salmon associations. These data provide a unique, extensive and solid baseline for monitoring parasite community structure in relation to environmental change.

Subchronic exposure to arsenic in rats was investigated to identify sensitive indicators of subclinical toxicity in rats. Immunological, pathological, and biochemical bioindicators were examined in rats exposed to arsenic in their drinking water. Juvenile male Wistar rats were allocated to four treatment groups receiving 0, 0.4, 4, and 40 ppm of arsenite in drinking water for 18 wks. Besides daily monitoring for clinical signs of adverse health effects, clinical biochemistry, B-cell-mediated and innate immune responses, plus gross, and histopathology were examined. In vitro tests of oxidative damage to basic cellular constituents, lipids, proteins, and nucleic acids, were measured using thiobarbituric acid reacting substances (TBARS) assays, protein carbonyl formation, and 8-hydroxydeoxyguanosine (8-OHdG), respectively. Clinical changes in the rats were limited to decreased feed and water intake in the high- (40 ppm) dose group (P < 0.05), however, growth rate was not affected. Serum biochemical changes occurred in blood urea nitrogen, K(+) , Cl(-) , and alanine aminotransferase (ALT) from arsenic exposure. Immunotoxicity was evident through a dose-dependent suppression of the secondary antibody-mediated response to a T-cell-dependent antigen, keyhole limpet hemocyanin (KLH). Histopathology of the liver revealed marked fatty infiltration and vacuolization particularly evident in periacinar hepatocytes. This pattern of toxicopathology in the high-exposure group may be related to the significantly higher (P < 0.05) oxidative stress, demonstrated through lipid peroxidation (TBARS assay) in the rats exposed to 40 ppm arsenite. The present study revealed that young, growing rats exposed to arsenic for 18 wks tolerated exposures up to 4 ppm. At higher doses, there was evidence of hepatotoxicity, humoral immunity was compromised, and an adverse effect on hepatic organelle and cell membranes was evident through a dose dependent increased in oxidative stress., (Copyright © 2010 Wiley Periodicals, Inc.)

Mercury (Hg) exposure was investigated through feathers of Olrog's Gull and related to health parameters in adults (hematocrit, total plasma proteins, morphometric measures, sex) and chicks (hematocrit, total plasma proteins, immunoglobulins G and M) from a colony located in estuary of Bahía Blanca, Argentina. Mercury concentrations were 5.50 ± 2.59 μg g⁻¹ (n = 44) in live adults, 1.85 ± 0.45 μg g⁻¹ (n = 45) in live chicks and 1.81 ± 0.41 μg g⁻¹ (n = 41) in dead chicks. Large differences were observed between live adults and live or dead chicks and small differences between live and dead chicks. In the adults, the sex of the birds was the variable that best explained Hg concentrations. Male birds had higher concentrations than females; this suggests that the clutch provides a sink for mercury during egg laying. Hg concentrations in both adults and live chicks were associated with higher hematocrits. This could be associated with upregulated erythropoiesis to compensate for increased rate of destruction of prematurely senescent, Hg-contaminated erythrocytes. Based on our results, on the levels of Hg pollution in the past in the study area, and on the dietary specialization of Olrog's Gull, we must be vigilant about potential negative effects of Hg pollution on this population and recommend continued monitoring on this threatened species.

This study established a disease model and protocol for bacterial challenge with Escherichia coli serogroup O2 strain EC317 in Japanese quail. Five groups of 10 birds each were injected subcutaneously in the breast with 200 μL of a brain-heart infusion (BHI) culture containing 1 × 10(8), 1 × 10(7), 1 × 10(6), 1 × 10(5), or 1 × 10(4) colony-forming units/mL of the test organism, which had been isolated from a turkey with cellulitis and septicemia. Birds in a 6th group were controls that received sterile BHI alone. Localized lesions of cellulitis developed in all of the birds that received E. coli. The morbidity and mortality rates were highest (100%) in the birds receiving the highest dose of E. coli and decreased linearly with decreasing dose (P < 0.05). Severity of disease, including lesions of pericarditis and perihepatitis, was also directly proportional to the dose of E. coli. These findings indicate that this disease challenge protocol can be used to study disease resistance and immunologic consequences of contaminant exposure or other stressors in birds.

The purpose of this study was to evaluate the impact of malathion on the immune system of wild birds, using Japanese quail (Coturnix coturnix japonica) as a model. Quail were exposed to malathion in drinking water at environmentally realistic concentrations (0 ppm, 1 ppm, and 10 ppm). In the fifth week, several arms of the immune response were tested using the T-cell based phytohemagglutinin (PHA) skin test, the B-cell mediated antibody response, and the chemiluminescence assay measuring innate immunity. After the sixth week of malathion exposure, quail were challenged with E. coli O2. The bursa of Fabricius and the spleen were assessed for histopathology. No clinical signs of malathion toxicity were observed. Morbidity or mortality subsequent to E. coli exposure tended (P = 0.08) to be higher in the high exposure group (50.0%) compared to the control (22.2%) group. There was no difference in the innate immune response in the malathion exposed birds, however, humoral immunity was suppressed (P = 0.03) with the higher malathion exposure. Histopathological evaluation revealed an immunosuppressive effect of malathion on the bursa of Fabricius; bursal atrophy, decreased B-cell density and increased apoptosis in the medulla, and increased connective tissue thickness of the follicular epithelium. Antibody suppression was correlated with bursal changes and peripheral blood lymphocyte count, the organ and cells involved in antibody production. Following the same pattern as other immunotoxicity tests, the PHA T-cell proliferative response also tended to be suppressed in the high exposure group. This study provides evidence that subchronic, moderate malathion exposure is immunotoxic to quail and that testing integrated, functional immunity using an infectious challenge is a better predictor of immunotoxicity than individual responses to immunotoxicity tests. The secondary antibody response, circulating lymphocyte populations, and bursal histopathology were the most sensitive indicators of immune status, as these predicted decreased disease resistance with malathion exposure.

The present study investigated the effects of lead (Pb) on immune responses in quail (Coturnix coturnix japonica) and the pathological impact of exposure to an infectious agent (E. coli O2). Fifty-four, 4-week-old quail were exposed to lead acetate in drinking water at 5 or 50 ppm. All birds were vaccinated with Newcastle Vaccine (NDV) during the third week of contaminant (Pb) exposure. In the fourth week, several arms of the immune response were tested using the T cell based phytohemagglutinin (PHA) skin test, the B cell mediated antibody response to NVD, and the chemiluminescence assay measuring innate immunity. Immunohistochemistry (IHC) was used to determine the expression of toll like receptor-3 (TLR-3) in the bursa of Fabricius. In the fifth week, quail were challenged with 200 μL of E. coli O2 (1×10(4) colony forming units (CFU)/mL). No clinical signs of Pb toxicity were observed. Morbidity/mortality subsequent to E. coli exposure was lowest in the high exposure group (27.8%) compared to low exposure (44.4%) and control (55.5%) groups. There was no difference in the T-cell-mediated PHA response, primary or secondary immune response or the innate response in Pb exposed groups; however, bursal TLR-3 increased (p<0.05) with higher Pb exposure. No evidence supported that subchronic Pb exposure was immunotoxic to quail at 5 or 50 ppm in drinking water. In contrast, our results provide evidence of a hormetic effect, with Pb exposed birds having lower morbidity and better survival than controls. Subchronic Pb exposure may be immunostimulatory rather than suppressive as predicted in earlier studies based on testing individual immune parameters., (Copyright © 2010 Elsevier Inc. All rights reserved.)

The wet landscape approach to oil sands tailings reclamation in the Athabasca Oil Sands region involves creating wetlands from fluid tailings in mined-out pits. We measured time to metamorphosis, thyroid hormone status, and detoxification enzyme (EROD) induction in Wood frog (Lithobates sylvaticus) tadpoles raised on reclaimed oil sands wetlands of different ages [young (≤ 7 yr) vs. old (> 7 yr)] and compared data with tadpoles raised on reference (control) wetlands. Metamorphosis was delayed or never occurred in tadpoles raised in young tailings; those exposed to older tailings developed similarly to those in reference wetlands. Thyroid hormone disruption likely played an important role in the metamorphosis delay as the T3:T4 ratio was lowest in tadpoles raised in young, tailings-affected wetlands. Our findings suggest tailings wetlands become less toxic with age, and that these amphibians will be able to complete their life cycle in tailing wetlands that have sufficiently detoxified with age., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Ranked among the top threats to conservation worldwide, infectious disease is of particular concern for wild canids because domestic dogs (Canis familiaris) may serve as sources and reservoirs of infection. On British Columbia's largely undeveloped but rapidly changing central and north coasts, little is known about diseases in wolves (Canis lupus) or other wildlife. However, several threats exist for transfer of diseases among unvaccinated dogs and wolves. To gain baseline data on infectious agents in this area, including those with zoonotic potential, we collected blood and stool samples from 107 dogs in 5 remote communities in May and September 2007. Serology revealed that the dogs had been exposed to canine parvovirus, canine distemper virus, Bordetella bronchiseptica, canine respiratory coronavirus, and Leptospira interrogans. No dogs showed evidence of exposure to Ehrlichia canis, Anaplasma phagocytophilum, Borrelia burgdorferi, Dirofilaria immitis, or Cryptococcus gattii. Of 75 stool samples, 31 contained at least 1 parasitic infection, including Taeniid tapeworms, the nematodes Toxocara canis and Toxascaris leonina, and the protozoans Isospora sp., Giardia sp., Cryptosporidium sp., and Sarcocystis sp. This work provides a sound baseline for future monitoring of infectious agents that could affect dogs, sympatric wild canids, other wildlife, and humans.

In the Athabasca oil sands region of northern Alberta, mining companies are evaluating reclamation using constructed wetlands for integration of tailings. From May to July 2008, reproductive performance of 40 breeding pairs of tree swallows (Tachycineta bicolor), plus growth and survival of nestlings, was measured on three reclaimed wetlands on two oil sands leases. A subset of nestlings was examined for i) feather corticosterone levels, ii) delayed-type hypersensitivity response, and iii) innate immune function. Nestlings on one of two wetlands created with oil sands process affected material (OSPM) were heavier and had greater wing-lengths, and mounted a stronger delayed-type hypersensitivity response compared those on the reference wetland. Corticosterone was significantly higher in male nestlings on one of two OSPM-containing wetland compared to the reference wetland. Body condition of 12-day-old female nestlings was inversely related to feather corticosterone. Under ideal weather conditions, reclaimed wetlands can support healthy populations of aerially-insectivorous birds., (Copyright (c) 2009 Elsevier Ltd. All rights reserved.)

First-stage nematode larvae with a dorsal-spine (DSL) were detected in five of 1,565 fecal samples from gray wolves (Canis lupus) collected in British Columbia, Canada, between 2005 and 2008. Molecular techniques were used to identify the DSL because it was not possible to determine their species identity using morphologic characters. The DSL were identified as Parelaphostrongylus odocoilei based on the results of single-strand conformation polymorphism (SSCP) analyses and DNA sequencing of the ribosomal DNA first and second internal transcribed spacers. Finding DSL of P. odocoilei in the feces of gray wolves was unexpected because P. odocoilei adults are parasites of cervids and bovids. The most likely explanation for the presence of DSL in wolf feces is that they were ingested along with the viscera of recently consumed prey. This was probably black-tailed deer (Odocoileus hemionus columbianus), which are known in the sampling area to be hosts of P. odocoilei. The present study demonstrates the use of SSCP and DNA sequencing for the identification, to the species level, of parasitic nematode larvae in feces.

Unlabelled: 1., Objectives: To validate the whole blood chemiluminescence (WBCL) assay in chickens, a simple and rapid method of measuring production of reactive oxygen species by circulating polymorphonuclear (PMN) leukocytes. To determine the physiological response and innate immune response associated with oral challenge with Clostridium perfringens in broiler chickens under different nutritional conditions. 2. In Experiment 1, birds were orally challenged with C. perfringens 1. type A or sham-challenged saline on days 14-21 post-hatch and fed protein-balanced diets containing 160 or 180 g crude protein/kg and 0.98 or 1.75% glycine in a 2 x 2 x 2 factorial arrangement of treatments. 2. Challenged birds had higher WBCL responses and more severe intestinal lesions than unchallenged birds. Birds fed diets containing 1.75% glycine had more intestinal lesions than those fed 0.98% glycine. 3. In Experiment 2 birds were fed protein-balanced diets containing 0.76, 2.10, 3.43 or 4.77% glycine. The birds fed 0.76% glycine diet had lower WBCL responses compared to birds fed the other three diets. Intestinal lesions were worse in the birds fed the highest, 4.77% glycine diet than in those fed the 0.76 or 2.10% glycine diets. 4. We conclude that the WBCL assay is a practical and sensitive means of assessing innate immune function in birds. The results suggest that both bacterial challenge and glycine content of chickens' diet influence their lesion scores and innate immune function.

One theory proposed to explain the global declines in amphibian populations involves contaminant-induced immune alteration and subsequent increased susceptibility to infectious disease. The goal of this study was twofold, to (1) study acute oral toxicity of deltamethrin (cyclopropanecarboxylic acid, 3-(2,2-dibromoethenyl)-2,2-dimethyl cyano(3-phenoxyphenyl)methyl ester) in tiger salamanders (Ambystoma tigrinum), and (2) evaluate whether the insecticide deltamethrin produces immunosuppression in these animals. In the acute toxicity study, tiger salamanders receiving single doses of deltamethrin ranging from 1 to 35 mg/kg displayed intention tremors, hypersalivation, ataxia, choreoathetosis (writhing), severe depression (immobility with minimal response to stimuli), and death. For acute effects, based on clinical signs, the median lethal dose (LD(50)) and lowest observed adverse effect level (LOAEL) were estimated to be 5 to 10 mg/kg and 1 mg/kg, respectively. The LOAEL in animals dosed 3 times per week for 4 wk was 400 microg/kg/d. The endpoints for the immunotoxicity study included lymphoid organ mass and histopathology, hematological variables, and functional assays of phagocytosis, oxidative burst, and lymphoblastic transformation. Tiger salamanders in 4 treatment groups (0, 4, 40, or 400 microg/kg/d) were dosed with deltamethrin via the diet 3 times per week for 4 wk. Deltamethrin exposure resulted in increased liver mass, packed cell volume, and total plasma protein concentration, but these effects were not dose dependent. The relative mass of kidney and spleen, plasma albumin and globulin concentrations, and circulating leukocyte numbers were not affected by deltamethrin exposure, nor were phagocytosis, oxidative burst, and lymphoblastic transformation. This study shows that at moderate levels of exposure, deltamethrin may be neurotoxic to tiger salamanders. However, based on the immune assays considered in this study there was no evidence of immunosuppression from dietary exposure to environmentally relevant concentrations of deltamethrin. In light of these findings, it is unlikely that exposure to environmental concentrations of deltamethrin has produced immunosuppression and contributed to the emergence of iridovirus outbreaks in tiger salamander populations.

Avian research involving examination of immune function or testing of immunocompetence in wild birds has been based upon information on Galliforms, (chicken and quail) even though they are precocial, whereas most wild species with which ecologists, biologists and toxicologists work are altricial; blind, naked and completely dependent at hatching. Here we begin to address this gap in knowledge, offering insight into the early, post-hatching, humoral immune response in an altricial bird, the American kestrel (Falco sparverius). Over two breeding seasons, nestling kestrels were immunized with a non-pathogenic antigen, dinitrophenol keyhole limpet hemocyanin (DNP-KLH), between 3 and 9 days post-hatching and boostered 6 days later. Background levels, primary and secondary immune responses were measured using an enzyme linked immunosorbent assay. The specificity of our laboratory produced rabbit, anti-kestrel antibody was determined using a double immunodiffusion assay. Results showed the rabbit antiserum to have specific anti-kestrel IgG activity. Birds as young as three days old could successfully mount an antibody response, the magnitude of which increased with age at first vaccination. Early immunization did not compromise growth rate, nor did it affect the maximum secondary response. Comparatively, adult kestrels immunized during the same season and following the same protocol, had antibody levels four times higher than those of the nestlings.

Environmental risk assessments are broadening to include evaluations of avian species exposed to gaseous and particulate materials (Mineau, 2002b; Irvine, 2004; Carmalt, 2005). Since the avian respiratory tract is fundamentally different from the respiratory tract of rodents, the effects of gaseous materials on birds cannot validly be extrapolated from data derived from rodent exposure studies (Briant & Driver, 1992; Brown et al., 1997). To address the lack of avian-specific lowest observable effect levels used to calculate reference concentrations for airborne pollutants, a system was designed to facilitate research on inhalation toxicology in small birds. Birds have long been used as early indicators of poor air quality (Brown et al., 1997), and various chambers have been designed for head only exposures of larger birds (Briant & Driver, 1992). Smaller birds with short tracheal lengths and hooked beaks however require less restrictive exposure apparatus, thus warranting the proposed design. The chamber described in this article was designed to accommodate a small falcon, the American kestrel, a species frequently used in toxicological risk assessments (Wiemeyer & Lincer, 1987a; Smits & Bortolotti, 2001; Bortolotti et al., 2003; Fisher et al., 2006). To accomplish this, a 41-L closed inhalation system capable of exposing 12 adult American kestrels was constructed primarily of galvanized steel, polyvinyl chloride, and copper tubing. Humidified air was passed over the birds and subsequently decontaminated by an activated carbon filter and released to a HEPA filtration system. The proposed inhalation chamber was successfully used in 2005 and 2006 to expose a total of 55 male American kestrels to benzene and toluene. Measurements of various biochemical endpoints associated with benzene and toluene toxicity allowed us to study the effects of airborne pollutants on small nondomesticated birds in a controlled laboratory setting.

Benzene and toluene are representative volatile organic compounds (VOC) released during production, storage, and transportation associated with the oil and gas industry and are chemicals of concern, as they are released in greater and possibly more biologically significant concentrations than other compounds. Most studies of air pollution in high oil and gas activity areas have neglected to consider risks to birds, including top-level predators. Birds can be used as highly sensitive monitors of air quality and since the avian respiratory tract is physiologically different from a rodent respiratory tract, effects of gases cannot be safely extrapolated from rodent studies. Wild and captive male American kestrels were exposed for approximately 1 h daily for 28 d to high (rodent lowest-observed-adverse-effect level [LOAEL] of 10 ppm and 80 ppm, respectively) or environmentally relevant (0.1 ppm and 0.8 ppm, respectively) levels of benzene and toluene. Altered immune responses characteristic of those seen in mammalian exposures were evident in kestrels. A decreased cell-mediated immunity, measured by delayed-type hypersensitivity testing, was evident in all exposed birds. There was no effect on humoral immunity. Plasma retinol levels as measured by high-performance liquid chromatography (HPLC) analysis were decreased in wild and captive kestrels exposed to the rodent LOAEL for combined benzene and toluene. This study indicates that American kestrels are sensitive to combined benzene and toluene. The study also illustrates the need for reference concentrations for airborne pollutants to be calculated, including sensitive endpoints specific to birds. Based on these findings, future studies need to include immune endpoints to determine the possible increased susceptibility of birds to inhaled toxicants.

The whole-blood chemiluminescence (WBCL) assay is a simple and rapid method of measuring production of reactive oxygen species by circulating leukocytes, particularly heterophils (birds) and neutrophils (other vertebrates). In the interest of substantiating a broadly applicable measure of innate immunity, we investigated the microplate WBCL method for several wildlife species as well as domestic broiler chickens. Lucigenin as a light enhancer was used for all avian blood, wild and domestic, while luminol was used in bear and frog blood. Use of ethylenedinitrilotetraacetic acid (EDTA) as the anticoagulant caused hemolysis of frog blood and decreased WBCL responses in all animals tested. Heparin, even in high concentrations, caused modest to no decrease of WBCL responses. The WBCL response correlated highly with heterophil or neutrophil numbers in all species tested. The WBCL response was tested in freshly collected blood as well as in blood one to five days postcollection in order to determine the utility of this assay for field studies when immediate access to laboratory facilities is not possible. One to three days of delay in performing the test after blood collection caused no, or only a slight, decay of the chemiluminescence response in most animals. Using domestic chickens, we tested the sensitivity of the WBCL method to detect differences between treatment groups and looked for loss of chemiluminescence response over several days using the original blood samples. Significant differences in the WBCL response between experimental groups of broiler chicken were detectable in freshly collected blood, as well as one- to four-day-old blood. Our results show that the innate immune response of populations of wild animals may be successfully compared using this assay, even when blood cannot be tested until a few days after collection.

The oil sands of Alberta, Canada are one of the world's largest reserves of crude oil. Oil sands mining companies are now investigating the ecological impacts of reclamation strategies in which wetlands are used for the bioremediation of waste materials. To examine the endocrine disrupting potential of chemicals in Oil Sands Process Materials (OSPM), thyroid hormone concentrations were measured in plasma and thyroid glands of nestling tree swallows (Tachycineta bicolor) from wetlands partly filled with mine tailings. Plasma triiodothyronine (T(3)) concentrations and thyroxine (T(4)) content within thyroid glands were elevated in nestlings from OSPM sites compared to those from the reference site. Results suggested enhanced hormone synthesis by the thyroid glands independently of activation of the pituitary-thyroid axis, as well as increased deiodination of T(4) into T(3) in peripheral tissues. This might have resulted from exposure to oil sands associated chemicals such as polycyclic aromatic hydrocarbons and from environmental factors such as food availability. Modulation of thyroid function might have negative effects on metabolism, behavior, feather development, and molt, which could compromise postfledging survival.

Naphthenic acids (NAs) are a group of carboxylic acids that are of particular concern to the steadily growing oil sands mining industry of Alberta, Canada, because they become highly concentrated in the water used for oil sands extraction and are toxic to aquatic biota and mammals. Upon mine closure, vast amounts of process-affected water will need to be reclaimed and proven safe for wildlife colonizing reclaimed areas. The effects of exposure to NAs have not been investigated in avian species. To address this void, tree swallow (Tachycineta bicolor) nestlings were dosed with NAs while being reared normally by their free-ranging parents on a site in the vicinity of the oil sands. Nestlings received 1.5 mg NAs/day (approximately 0.075 g/kg body mass) from d 7 to d 13 of age, which represented a 10-fold "worst exposure" scenario. Nestling growth, hematocrit, blood biochemistry, organ weights, and ethoxyresorufin O-deethylase (EROD) activity were unaffected by NAs. The only change detected on histopathological evaluation of major organs was an increase in extramedullary erythropoiesis in the liver. These findings indicate that nestling tree swallows can successfully tolerate short-term exposures to environmentally realistic concentrations of NAs. However, this study did not investigate the chronic or reproductive toxicity of NAs. More research needs to be conducted to complete this initial assessment, to determine environmental risks on reclaimed areas where birds will be breeding and where their exposure to NAs could extend for several weeks.

Oil sands mining is steadily expanding in Alberta, Canada. Major companies are planning reclamation strategies for mine tailings, in which wetlands will be used for the bioremediation of water and sediments contaminated with polycyclic aromatic hydrocarbons and naphthenic acids during the extraction process. A series of experimental wetlands were built on companies' leases to assess the feasibility of this approach, and tree swallows (Tachycineta bicolor) were designated as upper trophic biological sentinels. From May to July 2004, prevalence and intensity of infestation with bird blow flies Protocalliphora spp. (Diptera: Calliphoridae) were measured in nests on oil sands reclaimed wetlands and compared with those on a reference site. Nestling growth and survival also were monitored. Prevalence of infestation was surprisingly high for a small cavity nester; 100% of the 38 nests examined were infested. Nests on wetlands containing oil sands waste materials harbored on average from 60% to 72% more blow fly larvae than those on the reference site. Nestlings on reclaimed sites suffered mean parasitic burdens about twice that of those on the reference site; and for comparable parasitic load, they exhibited greater pathologic effects (e.g., decreased body mass) than control nestlings. The heavy blow fly infestation on oil sands-impacted wetlands suggests that oil sands mining disturbs several components of the local ecosystem, including habitat characteristics, blow fly predators, and host resistance to parasites.

Tree swallows (Tachycineta bicolor) are widely used as indicators of local polychlorinated biphenyl (PCB) contamination in North America. Although determining total PCB residues in tissues is useful in environmental monitoring, analysis of PCB congener profiles may reveal sources of contamination and thus prove to be a more refined tool to track contaminants through the food web. To show how differences in PCB congener patterns in birds can be linked to the PCB patterns in their prey, we evaluated PCB congeners in tissues of tree swallow nestlings and their insect prey using principal component analysis and Euclidean similarities. The PC1 scores for PCB residues in nestlings fell between those of the Hexagenia (Ephemeroptera, mayflies) and Chironomidae (Diptera, midges), the two major prey groups of the nestlings. The congener pattern was not related to the location of nest boxes within the study area. However, Hexagenia insects and the nestlings that consumed them were richer in less chlorinated congeners and had higher PC1 scores than Chironomidae insect. In concordance, congener pattern of nestlings that hatched earlier and consumed more mayflies was more similar than that of other nestling to the pattern of nestings texagenia as calculated by Euclidean similarities. We point to the importance of understanding the seasonal availability of specific types of insect prey and their PCB congener pattern before these data are applied in models of trophic transfer of individual PCB congeners within a food web.

In 1998, the Aznalcóllar mine tailings dyke in southwestern Spain broke, flooding the Agrio-Guadiamar river system with acid tailings up to the borders of one of the largest breeding colonies of white storks in the western Palearctic, Dehesa de Abajo. Over the following years, a high proportion of nestlings developed leg defects not seen before the spill, prompting this study. Nestlings with deformed legs had significantly lower plasma phosphorous (P) and higher Ca:P ratios than non-deformed cohorts in the first two years, but in the third year, when more, younger birds were studied, plasma P ranged from much higher to much lower in the affected colony compared with reference birds. Coefficients of variation for phosphorous were 19% and 60%, in reference and contaminated colonies, respectively. Storks from the contaminated colony were unable to control P levels and Ca:P ratios within the narrow limits necessary for normal bone development.

Captive breeding (n = 25 pairs) and nonbreeding (n = 25) American kestrels were exposed to a mixture of polychlorinated biphenyls (PCBs) (Aroclor 1248:1254:1260) through their diet of day-old cockerels. Kestrels ingested approximately 7 mg/kg body weight each day of PCBs, and this dosage resulted in environmentally relevant total PCB residues in eggs (geometric mean of 34.1 microg/g). An equal number of unexposed birds served as controls. Bare areas of skin known as brood patches function during incubation to warm eggs; therefore, brood patch size could potentially influence hatching success, or patches may be a confounding factor in the relationship between poor incubation behavior and hatching failure observed in birds in toxicological studies. Exposure to PCBs altered the size of brood patches in American kestrels. PCB-exposed male and female nonbreeders had two of three brood patches that were larger than those of control nonbreeders. Breeding males exposed to PCBs had smaller patches than controls, whereas PCB-exposed female kestrels had one larger and one smaller patch than controls. Patch sizes were not related to total PCB residue levels in eggs of exposed birds. Brood patches were not related to various incubation behaviors or hatching success in either control or PCB-exposed kestrels.

Oil sands mining companies in Alberta, Canada, are evaluating the feasibility of using wetlands to detoxify oil sands process material (OSPM) as a reclamation strategy. Reproductive success, nestling growth, survival and ethoxyresorufin-o-deethylase (EROD) activity were measured in tree swallows (Tachycineta bicolor) on experimental wetlands. In 2003, harsh weather triggered a widespread nestling die-off. Mortality rates on the control site reached 48% while they ranged from 59% to 100% on reclaimed wetlands. The odds of dying on the most process-affected sites were more than ten times higher than those on the control site. In 2004, weather was less challenging. Mortality rates were low, but nestlings on reclaimed wetlands weighed less than those on the control site, and had higher EROD activity. These results indicate that compared with reference birds, nestlings from OSPM-impacted wetlands may be less able to withstand additional stressors, which could decrease their chances of survival after fledging.

We investigated whether polychlorinated biphenyl (PCB) exposure in American kestrels (Falco sparverius) influenced incubation behavior and whether altered behavior could lead to poor reproductive success. Captive kestrels were fed a mixture of PCBs (Aroclors 1248:1254:1260) at an approximate daily dose of 7 mg/kg body weight, 1 month prior to pairing and throughout incubation. Behaviors of 23 control and 23 PCB-exposed pairs were monitored throughout incubation using an electronic balance in the nest box. PCB exposure resulted in longer incubation periods and in altered incubation behaviors. Seven of 14 behavioral variables showed some association with treatment, with sex-specific effects largely biased toward disrupted male behavior. For most behaviors, the treatment effect was explained by the delayed clutch initiation induced by PCBs rather than by a direct physiological impact of the contaminants. PCB-exposed pairs with greater attendance to their eggs and better coordination of incubation duties had improved hatching success.

Point Pelee National Park of Canada in southwestern Ontario, an important migratory route and vital breeding area for many birds, has localized areas of organochlorine (OC) pesticide contamination from agricultural production during the 1950s and 1960s. During 2001 and 2002, we investigated movement of persistent contaminants through the food web with the insectivorous tree swallow (Tachycineta bicolor) as a sentinel. The a priori site classifications, contaminated or reference, were based on soil residues of dichlorodiphenyltrichloroethane (DDT) and its breakdown products (sigmaDDT), dieldrin, and other OC pesticides. In 2001, all nestling tissue samples were pooled by site, and residue levels did not reflect the soil contaminant status. To improve sampling accuracy in 2002, tissue residues were determined from birds in individual nests. This showed OC pesticides to be higher in samples from contaminated sites compared with reference sites (p = 0.031). Polychlorinated biphenyls (PCBs), which were not detected in soil samples, were present in the nestlings and were not related to site of origin (p = 0.422). In 2002, dietary samples were collected from nestlings and identified to taxon, and representative insects collected from nesting sites were analyzed for PCBs and other OCs. Consumption of terrestrial prey was positively correlated with tissue residues of sigmaDDT (p = 0.006), whereas PCBs came from aquatic prey, Hexagenia mayflies (p = 0.003). Dietary details proved valuable in this study of contaminant transfer in insectivorous vertebrates.

Point Pelee National Park in southwestern Ontario, Canada--a major migratory route and vital breeding area for many birds--has localized areas of organochlorine (OC) contamination from the 1950s and 1960s. During 2002, we investigated the effect of tissue OC contaminant levels on the physiology and growth of nestling tree swallows (Tachycineta bicolor). Polychlorinated biphenyl (PCB) contamination in the nestlings positively correlated with hepatic ethoxy-, benzyloxy-, and pentoxyresorufin-o-dealkylase enzyme activities and liver size. Despite detectable physiologic changes associated with body burdens of PCBs, reproductive success of breeding pairs was not affected. Hatching date was negatively correlated with PCB levels, alkoxyresorufin-O-dealkylase (alkROD) activities, liver size, and serum protein levels. alkROD activities were largely dependent on hatching date because insect prey contaminated with PCBs were only abundant during a limited period of time during the breeding season.

Identifying the potential effects of industrially formed wetlands on waterfowl populations is important for assessing the suitability of such wetlands in industrial reclamation strategies. Mallard ducklings were held in situ on two industrially formed wetlands and one reference wetland in northern Alberta, Canada. Duckling mass and skeletal size were measured at regular intervals over 33 d, and blood was collected to investigate the analysis of plasma metabolites (triglyceride and glycerol) as an indicator of physiological condition. In repeated-measures analysis of variance (ANOVA), multivariate ANOVA, and subsequent multiple-comparisons tests, body mass and skeletal size were significantly lower in ducklings maintained on the industrial wetland after 2, 5, 9, and 13 d of exposure. In this situation, plasma metabolite analysis did not provide additional information on mass-independent condition. We conclude that if the observed differences in growth and size translate into a decreased survival of juvenile waterfowl inhabiting these wetlands, then populations of these birds in the area could be negatively affected. We emphasize the importance of field-based ecological research in toxicological studies of wildlife.