REFERENCE VALUES AND COMPARISON OF BLOOD CHEMISTRY AND PLASMA PROTEIN VALUES BETWEEN GOLD STANDARD ANALYZERS AND FOUR POINT-OF-CARE DEVICES IN FREE-RANGING CANVASBACKS (AYTHYA VALISINERIA).

Accurate, timely, and cost-effective blood chemistry analysis is an essential tool for directing emergency treatment, monitoring the health status of captive and free-ranging individuals and flocks, and improving the efficacy of conservation actions. Blood samples were obtained from 52 canvasbacks (Aythya valisineria) that were captured on San Francisco Bay, California, during December 2017 as part of a long-term study. Reference values and clinical agreement were determined for blood chemistry and plasma protein parameters among four commonly used point-of-care devices (VetScan® VS2, i-STAT®, AlphaTRAK®2 glucometer, refractometer) and two gold standard laboratory analyzers (Roche cobas® c501, Helena SPIFE 3000 system). Canvasback reference values were generally within expected ranges for Anatidae species with the exception of higher upper limits for sodium and chloride. Creatine kinase and aspartate transaminase values exceeded a published threshold for diagnosis of capture myopathy even though study birds were captured using low-stress techniques and successfully released. With the exception of higher alkaline phosphatase in hatch-year canvasbacks, no age or sex differences were observed for any analyte in this population that was captured during a nonbreeding period. Analysis of analyzer agreement found raw VetScan aspartate transaminase, calcium, glucose, and uric acid values; corrected VetScan albumin, potassium, sodium, and total protein values; raw i-STAT glucose and potassium values; and corrected i-STAT sodium and chloride values were clinically interchangeable with Roche cobas values. Raw VetScan and i-STAT glucose values were also interchangeable. However, none of the Roche or point-of-care analyzer plasma protein values were in clinical agreement with gold standard electrophoresis values. The findings of this study highlight the need for analyzer- or technique-specific reference values and provide biologists and veterinarians quantitative reference values using currently available analyzers to better assess and respond to the health of individuals and populations.

Early detection of wildlife morbidity and mortality through an event-based surveillance system.

The ability to rapidly detect and respond to wildlife morbidity and mortality events is critical for reducing threats to wildlife populations. Surveillance systems that use pre-diagnostic clinical data can contribute to the early detection of wildlife morbidities caused by a multitude of threats, including disease and anthropogenic disturbances. Here, we demonstrate proof of concept for use of a wildlife disease surveillance system, the 'Wildlife Morbidity and Mortality Event Alert System', that integrates pre-diagnostic clinical data in near real-time from a network of wildlife rehabilitation organizations, for early and enhanced detection of unusual wildlife morbidity and mortality events. The system classifies clinical pre-diagnostic data into relevant clinical classifications based on a natural language processing algorithm, generating alerts when more than the expected number of cases is recorded across the rehabilitation network. We demonstrated the effectiveness and efficiency of the system in alerting to events associated with both common and emerging diseases. Tapping into this readily available unconventional general surveillance data stream offers added value to existing wildlife disease surveillance programmes through a relatively efficient, low-cost strategy for the early detection of threats.

A review of the toxicology of oil in vertebrates: what we have learned following the Deepwater Horizon oil spill.

In the wake of the Deepwater Horizon (DWH) oil spill, a number of government agencies, academic institutions, consultants, and nonprofit organizations conducted lab- and field-based research to understand the toxic effects of the oil. Lab testing was performed with a variety of fish, birds, turtles, and vertebrate cell lines (as well as invertebrates); field biologists conducted observations on fish, birds, turtles, and marine mammals; and epidemiologists carried out observational studies in humans. Eight years after the spill, scientists and resource managers held a workshop to summarize the similarities and differences in the effects of DWH oil on vertebrate taxa and to identify remaining gaps in our understanding of oil toxicity in wildlife and humans, building upon the cross-taxonomic synthesis initiated during the Natural Resource Damage Assessment. Across the studies, consistency was found in the types of toxic response observed in the different organisms. Impairment of stress responses and adrenal gland function, cardiotoxicity, immune system dysfunction, disruption of blood cells and their function, effects on locomotion, and oxidative damage were observed across taxa. This consistency suggests conservation in the mechanisms of action and disease pathogenesis. From a toxicological perspective, a logical progression of impacts was noted: from molecular and cellular effects that manifest as organ dysfunction, to systemic effects that compromise fitness, growth, reproductive potential, and survival. From a clinical perspective, adverse health effects from DWH oil spill exposure formed a suite of signs/symptomatic responses that at the highest doses/concentrations resulted in multi-organ system failure.

REVIEW: Assessing North Atlantic right whale health: threats, and development of tools critical for conservation of the species.

Whaling has decimated North Atlantic right whales Eubalaena glacialis (NARW) since the 11th century and southern right whales E. australis (SRW) since the 19th century. Today, NARWs are Critically Endangered and decreasing, whereas SRWs are recovering. We review NARW health assessment literature, NARW Consortium databases, and efforts and limitations to monitor individual and species health, survival, and fecundity. Photographs are used to track individual movement and external signs of health such as evidence of vessel and entanglement trauma. Post-mortem examinations establish cause of death and determine organ pathology. Photogrammetry is used to assess growth rates and body condition. Samples of blow, skin, blubber, baleen and feces quantify hormones that provide information on stress, reproduction, and nutrition, identify microbiome changes, and assess evidence of infection. We also discuss models of the population consequences of multiple stressors, including the connection between human activities (e.g. entanglement) and health. Lethal and sublethal vessel and entanglement trauma have been identified as major threats to the species. There is a clear and immediate need for expanding trauma reduction measures. Beyond these major concerns, further study is needed to evaluate the impact of other stressors, such as pathogens, microbiome changes, and algal and industrial toxins, on NARW reproductive success and health. Current and new health assessment tools should be developed and used to monitor the effectiveness of management measures and will help determine whether they are sufficient for a substantive species recovery.

Effects of Season, Location, Species, and Sex on Hematologic and Plasma Biochemical Values and Body Mass in Free-ranging Grebes (Aechmophorus species).

The effects of season, location, species, and sex on body weight and a comprehensive array of blood chemistry and hematology analytes were compared for free-ranging western (Aechmophorus occidentalis) and Clark's (Aechmophorus clarkii) grebes. Birds (n = 56) were collected from Puget Sound, WA, and Monterey Bay and San Francisco Bay, CA, from February 2007 to March 2011. The data supported generalization of observed ranges for most analytes across Aechmophorus grebe metapopulations wintering on the Pacific coast. Notable seasonal and location effects were observed for packed cell volume (winter 6% greater than fall; winter California [CA] 5% greater than Washington [WA]), total white blood cell count (CA 3.57 × 103 cells/µL greater than WA), heterophils (WA 10% greater than CA), lymphocytes (winter 19% greater than fall), heterophil to lymphocyte ratio (fall 5.7 greater than winter), basophils (CA greater than WA), plasma protein (WA about 10 g/L [1.0 g/dL] greater than CA), plasma protein to fibrinogen ratio (winter about 15 greater than fall), potassium (CA 2 mmol/L greater than WA), and liver enzymes (alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase: WA greater than CA). Within California, season had a greater effect on body mass than sex (mean winter weights about 200 g greater than fall), whereas within a season, males weighed only about 80 g more than females, on average. These data give biologists and veterinarians quantitative reference values to better assess health at the individual and metapopulation level.

Monitoring of fungal loads in seabird rehabilitation centers with comparisons to natural seabird environments in northern California.

Aspergillosis remains a major cause of mortality in captive and rehabilitated seabirds. To date, there has been poor documentation of fungal (particularly Aspergillus spp.) burdens in natural seabird loafing and roosting sites compared with fungal numbers in rehabilitation or captive settings and the various microenvironments that seabirds are exposed to during the rehabilitation process. This study compares fungal, particularly Aspergillus spp., burdens potentially encountered by seabirds in natural and rehabilitation environments. Differences among the various microenvironments in the rehabilitation facility were evaluated to determine the risk of infection when seabirds are experiencing high stress and poor immune function. Aspergillus spp. counts were quantified in three wildlife rehabilitation centers and five natural seabird loafing and roosting sites in northern California using a handheld impact air sampler and a water filtration system. Wildlife rehabilitation centers demonstrated an increase in numbers of conidia of Aspergillus spp. and Aspergillus fumigatus in air and water samples from select aquatic bird rehabilitation centers compared with natural seabird environments in northern California. Various microenvironments in the rehabilitation facility were identified as having higher numbers of conidia of Aspergillus spp. These results suggest that periodic monitoring of multiple local areas, where the birds spend time in a rehabilitation facility, should be done to identify "high risk" sites, where birds should spend minimal time, or sites that should be cleaned more frequently or have improved air flow to reduce exposure to fungal conidia. Overall, these results suggest that seabirds may be more likely to encounter Aspergillus spp. in various microenvironments in captivity, compared with their native habitats, which could increase their risk of developing disease when in a debilitated state.

Molecular sub-typing suggests that the environment of rehabilitation centers may be a potential source of Aspergillus fumigatus infecting rehabilitating seabirds.

Aspergillosis remains a major cause of infection-related avian mortality in birds that are debilitated and undergoing rehabilitation for release into the wild. This study was designed to understand the source of avian aspergillosis in seabirds undergoing rehabilitation at selected northern California aquatic bird rehabilitation centers. Air, surface and water sampling was performed between August 2007 and July 2008 in three such centers and selected natural seabird loafing sites. Average air Aspergillus fumigatus counts were at least nine times higher in samples obtained from the rehabilitation sites (M = 7.34, SD = 9.78 CFU/m(3)), when compared to those found at natural sites (M = 0.76, SD = 2.24 CFU/m(3)), t (205) = -5.99, P < 0.001. A total of 37 A. fumigatus isolates from birds with confirmed aspergillosis and 42 isolates from environmental samples were identified using both morphological and molecular methods, and subsequently sub-typed using an eight-locus microsatellite panel with the neighbor joining algorithm. Results of the study demonstrated the presence of five clonal groups, 13 genotypically related clusters, and 59 distinct genotypes. Six of the 13 genotypically related clusters contained matching genotypes between clinical isolates and local environmental isolates from the rehabilitation center in which these birds were housed. We present evidence that the environment of rehabilitation centers may be a source for A. fumigatus infection in rehabilitated seabirds.

Evaluation of plasma (1-->3) beta-D-glucan concentrations in birds naturally and experimentally infected with aspergillus fumigatus.

Avian aspergillosis, most often caused by Aspergillus fumigatus, is a common and devastating disease affecting a range of bird species. Early diagnosis is difficult and often unreliable. The current study evaluated the utility of measuring (1-->3)-beta-D-glucan (BG) concentrations in avian plasma samples to aid in the diagnosis of aspergillosis. We evaluated a commercially available BG assay (Fungitell, Beacon Diagnostics) using 178 plasma samples from naturally infected, experimentally infected, and aspergillosis-free birds. Although there was variation in BG concentration, as reflected by high standard deviations, seabirds with confirmed aspergillosis had the highest mean BG concentrations (M = 3098.7 pg/dl, SD = 5022.6, n = 22) followed by companion avian species and raptors with confirmed aspergillosis (M = 1033.8 pg/dl, SD = 1531.6, n = 19) and experimentally infected Japanese quail (Coturnix japonica; M = 1066.5 pg/dl, SD = 1348.2, n = 17). Variation in severity of disease, differences among species of birds with and without disease, and also different levels in environmental exposure likely contribute to the differences among avian groups. The overall sensitivity and specificity of the BG test for diagnosis of aspergillosis in birds was 60.0 and 92.7%, respectively, with an overall optimized avian cut-offvalue of > or = 461 pg/dl for positive disease. Our findings suggest that, although BG concentrations are highly variable between and within different avian groups, it could serve as a useful adjunctive diagnostic test for aspergillosis that is applicable to multiple avian species in some settings, particularly as a negative predictor of infection.

Effects of freshwater housing and fluid types on aquatic bird serum electrolyte concentrations.

Recent seabird mass morbidity events have highlighted the need to elucidate the effects of freshwater and hypertonic saline use for fluid therapy and housing in captive aquatic birds. Serum electrolyte concentrations of northern fulmars (Fulmarus glacialis; n = 9), western grebes (Aechmophorus occidentalis; n = 6), and common murres (Uria aalge; n = 25) housed on freshwater while undergoing rehabilitation at the San Francisco Bay Oiled Wildlife Care and Education Center in Fairfield, California, were compared to reference intervals from free-ranging populations. Additionally, northern fulmars were given intermittent oral boluses of hypertonic saline. In birds housed on freshwater, there were significant decreases in serum sodium and chloride, with 44% of northern fulmars and 72% of common murres falling below the established reference interval. All of the western grebes were able to maintain serum sodium and chloride within the reference intervals. The significance of these findings reflect the behavior and natural history of the species studied. The physiologic regulation of salt by the salt glands, gastrointestinal tract, and renal systems of seabirds, along with their behavior and natural history, should be considered when working with these birds in a rehabilitation or captive setting. Salt supplementation is necessary for some species of seabirds. Monitoring of serum electrolytes should be used for individual animals or salt supplementation should be considered in holopelagic species.

Pathogen surveillance and epidemiology in endangered Peninsular bighorn sheep (Ovis canadensis nelsoni).

Peninsular bighorn sheep (Ovis canadensis nelsoni) are found exclusively in Southern California and Baja Mexico. They are federally endangered due to multiple threats, including introduced infectious disease. From 1981 - 2017, we conducted surveillance for 16 pathogens and estimated population sizes, adult survival, and lamb survival. We used mixed effects regression models to assess disease patterns at the individual and population levels. Pathogen infection/exposure prevalence varied both spatially and temporally. Our findings indicate that the primary predictor of individual pathogen infection/exposure was the region in which an animal was captured, implying that transmission is driven by local ecological or behavioral factors. Higher Mycoplasma ovipneumoniae seropositivity was associated with lower lamb survival, consistent with lambs having high rates of pneumonia-associated mortality, which may be slowing population recovery. There was no association between M. ovipneumoniae and adult survival. Adult survival was positively associated with population size and parainfluenza-3 virus seroprevalence in the same year, and orf virus seroprevalence in the previous year. Peninsular bighorn sheep are recovering from small population sizes in a habitat of environmental extremes, compounded by infectious disease. Our research can help inform future pathogen surveillance and population monitoring for the long-term conservation of this population.

Zoonotic pathogens isolated from wild animals and environmental samples at two California wildlife hospitals.

OBJECTIVE: To determine types and estimate prevalence of potentially zoonotic enteric pathogens shed by wild animals admitted to either of 2 wildlife hospitals and to characterize distribution of these pathogens and of aerobic bacteria in a hospital environment. DESIGN: Cross-sectional study. SAMPLE: Fecal samples from 338 animals in 2 wildlife hospitals and environmental samples from 1 wildlife hospital. PROCEDURES: Fecal samples were collected within 24 hours of hospital admission. Environmental samples were collected from air and surfaces. Samples were tested for zoonotic pathogens via culture techniques and biochemical analyses. Prevalence of pathogen shedding was compared among species groups, ages, sexes, and seasons. Bacterial counts were determined for environmental samples. RESULTS: Campylobacter spp, Vibrio spp, Salmonella spp, Giardia spp, and Cryptosporidium spp (alone or in combination) were detected in 105 of 338 (31%) fecal samples. Campylobacter spp were isolated only from birds. Juvenile passerines were more likely to shed Campylobacter spp than were adults; prevalence increased among juvenile passerines during summer. Non-O1 serotypes of Vibrio cholerae were isolated from birds; during an oil-spill response, 9 of 10 seabirds screened were shedding this pathogen, which was also detected in environmental samples. Salmonella spp and Giardia spp were isolated from birds and mammals; Cryptosporidium spp were isolated from mammals only. Floors of animal rooms had higher bacterial counts than did floors with only human traffic. CONCLUSIONS AND CLINICAL RELEVANCE: Potentially zoonotic enteric pathogens were identified in samples from several species admitted to wildlife hospitals, indicating potential for transmission if prevention is not practiced.

Short-term survival and effects of transmitter implantation into western grebes using a modified surgical procedure.

Two pilot trials and one study in a closely related grebe species suggest that Western grebes (Aechmophorus occidentalis) will not tolerate intracoelomic transmitter implantation with percutaneous antennae and often die within days of surgery. Wild Western grebes (n = 21) were captured to evaluate a modified surgical technique. Seven birds were surgically implanted with intracoelomic transmitters with percutaneous antennae by using the modified technique (transmitter group), 7 received the same surgery without transmitter implantation (celiotomy group), and 7 served as controls (only undergoing anesthesia). Modifications included laterally offsetting the body wall incision from the skin incision, application of absorbable cyanoacrylate tissue glue to the subcutaneous space between the body wall and skin incisions, application of a waterproof sealant to the skin incision after suture closure, and application of a piece of porcine small intestine submucosa to the antenna egress. Survival did not differ among the 3 groups with 7 of 7 control, 6 of 7 celiotomy, and 6 of 7 transmitter birds surviving the 9-day study. Experimental birds were euthanized at the end of the study, and postmortem findings indicated normal healing. Significant differences in plasma chemistry or immune function were not detected among the 3 groups, and only minor differences were detected in red blood cell indices and plasma proteins. After surgery, the birds in the transmitter group spent more time preening tail feathers than those in the control and celiotomy groups. These results demonstrate that, in a captive situation, celiotomy and intracoelomic transmitter implantation caused minimal detectable homeostatic disturbance in this species and that Western grebes can survive implantation of intracoelomic transmitters with percutaneous antennae. It remains to be determined what potential this modified surgical procedure has to improve postoperative survival of Western grebes that are intracelomically implanted with transmitters with percutaneous antennae and released into the wild.

Fishing gear-related injury in California marine wildlife.

We reviewed medical records from select wildlife rehabilitation facilities in California to determine the prevalence of injury in California Brown Pelicans (Pelecanus occidentalis), gulls (Larus spp.), and pinniped species (Zalophus californianus, Mirounga angustirostris, and Phoca vitulina) due to fishing gear entanglement and ingestion from 2001 to 2006. Of 9,668 Brown Pelican, gull, and pinniped cases described during the 6-yr study period (2001-06), 1,090 (11.3%) were fishing gear-related. Pelican injuries caused by fishing gear were most common in the Monterey Bay region, where 59.6% of the pelicans rescued in this area and admitted to a rehabilitation center were injured by fishing gear over the 6-yr period. The highest prevalence of fishing gear-related injury in gulls was documented in the Los Angeles/Orange County region (16.1%), whereas the highest prevalences in pinnipeds were seen in the San Diego region (3.7%). Despite these higher prevalences of gull and pinniped fishing gear-related injuries in these specific regions, there was no statistical significance in these trends. Juvenile gulls and pinnipeds were more commonly injured by fishing gear than adults (gulls: P = 0.03, odds ratio = 1.29; pinnipeds: P = 0.01, odds ratio = 2.07). Male pinnipeds were twice as likely to be injured by fishing gear as females (P < 0.01, odds ratio = 2.19). The proportion of fishing gear-related injury cases that were successfully rehabilitated and released (percentage of cases successfully rehabilitated to the point of release out of the total number of fishing gear-related injury cases) was high in all three species groups (pelicans: 63%; gulls: 54%; pinnipeds: 70%). Fishing gear-related injuries in Brown Pelicans and gulls were highest in the fall, but there was only a significant difference between seasons for fishing gear-related injuries in pelicans. Fishing gear-related injuries in pinnipeds most commonly occurred in summer; however, a statistical difference was not detected between seasons for pinnipeds. Derelict fishing gear-lost, abandoned or discarded sport and commercial line, nets, traps, etc.-in the marine environment is a significant cause of injury in California coastal marine wildlife. We evaluated data for stranded animals only; our results may underestimate the true number of coastal marine animals injured by lost or discarded fishing gear in California.

Use of plumage and gular pouch color to evaluate condition of oil spill rehabilitated California brown pelicans (Pelecanus occidentalis californicus) post-release.

Sublethal effects of oil spills may dampen seabird rehabilitation success due to lingering negative impacts of contamination and stress on reproduction and long-term survival. These effects can be difficult to measure while birds are in care as well as once birds are released. Expression of sexually selected traits that are sensitive to condition can provide information on physiological status of birds. We evaluated plumage molt and gular pouch skin color of California brown pelicans (Pelecanus occidentalis californicus) following oil contamination and rehabilitation to test for differences between previously oiled and rehabilitated (post-spill) and presumably uncontaminated pelicans. Post-spill pelicans released with either color leg bands alone, or bands plus harness-mounted satellite GPS tags, were relocated and visually assessed in the field at non-breeding communal roosts and compared to surrounding unmarked pelicans in the general population. Non-oiled pelicans bearing GPS tags were also included in the study. Post-spill pelicans lagged the general population in molt of ornamental yellow crown feathers but hind neck transition into white plumage was not significantly different. Both post-spill and non-oiled pelicans wearing GPS tags had lower gular redness scores than the unmarked, non-oiled population. Pre-breeding gular pouch redness of post-spill pelicans was more strongly influenced by wearing of a GPS tag than a history of oil contamination and rehabilitation. Gular pouch redness of post-spill pelicans in the first 18 months after release was positively correlated with long term survivorship. If gular pouch color is a condition-dependent sexual signal and overall health influences plumage molt progression, our results indicate that many post-spill pelicans marked with bands alone were in relatively good condition going into the next breeding season, but those released with electronic tags experienced additional stress due to wearing the equipment, introducing a confounding variable to the post-release study.

Health risks for marine mammal workers.

Marine mammals can be infected with zoonotic pathogens and show clinical signs of disease, or be asymptomatic carriers of such disease agents. While isolated cases of human disease from contact with marine mammals have been reported, no evaluation of the risks associated with marine mammal work has been attempted. Therefore, we designed a survey to estimate the risk of work-related injuries and illnesses in marine mammal workers and volunteers. The 17-question survey asked respondents to describe their contact with marine mammals, injuries sustained, and/or illnesses acquired during their period of marine mammal exposure. Most respondents, 88% (423/483), were researchers and rehabilitators. Of all respondents, 50% (243/483) reported suffering an injury caused by a marine mammal, and 23% (110/483) reported having a skin rash or reaction. Marine mammal work-related illnesses commonly reported included: 'seal finger' (Mycoplasma spp. or Erysipelothrix rhusiopathiae), conjunctivitis, viral dermatitis, bacterial dermatitis, and non-specific contact dermatitis. Although specific diagnoses could not be confirmed by a physician through this study, severe illnesses were reported and included tuberculosis, leptospirosis, brucellosis, and serious sequelae to seal finger. Risk factors associated with increased odds of injury and illness included prolonged and frequent exposure to marine mammals; direct contact with live marine mammals; and contact with tissue, blood, and excretions. Diagnosis of zoonotic disease was often aided by veterinarians; therefore, workers at risk should be encouraged to consult with a marine mammal veterinarian as well as a physician, especially if obtaining a definitive diagnosis for an illness becomes problematic.

AN EXPERIMENTAL STUDY OF THE EFFECTS OF CHEMICALLY DISPERSED OIL ON FEATHER STRUCTURE AND WATERPROOFING IN COMMON MURRES ( URIA AALGE).

Following an oil spill in the marine environment, chemical dispersants, which increase oil droplet formation and distribution into the water column, are assumed to provide a net benefit to seabirds by reducing the risk of exposure to oil on the water surface. However, few data are available regarding acute, external impacts of exposure to dispersed oil. We evaluated the effects of known concentrations of dispersant and crude oil in artificial seawater on live Common Murres ( Uria aalge). Waterproofing and microscopic feather geometry were evaluated over time and compared to pre-exposure values. Birds exposed to a high concentration of dispersant experienced an immediate, life-threatening loss of waterproofing and buoyancy, both of which resolved within 2 d. Birds exposed to oil, or a dispersant and oil mixture, experienced dose-dependent waterproofing impairment without resolution over 2 d. Alterations in feather geometry were observed in oil-exposed or dispersant- and oil-exposed birds and were associated with increased odds of waterproofing impairment compared to control birds. At a given contaminant concentration, there were no significant differences in waterproofing between oil-exposed and dispersant- and oil-exposed birds. We found that acute, external effects of oil and dispersed oil exposure are comparable and dose-dependent. Our results also indicate that a zero-risk assumption should not be used when seabirds are present within the dispersant application zone.

Mycobacterium avium subspecies paratuberculosis and Mycobacterium avium subsp. avium infections in a tule elk (Cervus elaphus nannodes) herd.

Between 2 August and 22 September 2000, 37 hunter-killed tule elk (Cervus elaphus nannodes) were evaluated at the Grizzly Island Wildlife Area, California, USA, for evidence of paratuberculosis. Elk were examined post-mortem, and tissue and fecal samples were submitted for radiometric mycobacterial culture. Acid-fast isolates were identified by a multiplex polymerase chain reaction (PCR) that discriminates among members of the Mycobacterium avium complex (MAC). Histopathologic evaluations were completed, and animals were tested for antibodies using a Johne's enzyme-linked immunosorbent assay (ELISA) and agar gel immunodiffusion. In addition, 104 fecal samples from tule elk remaining in the herd were collected from the ground and submitted for radiometric mycobacterial culture. No gross lesions were detected in any of the hunter-killed animals. Mycobacterium avium subsp. paratuberculosis (MAP) was cultured once from ileocecal tissue of one adult elk and was determined to be a strain (A18) found commonly in infected cattle. One or more isolates of Mycobacterium avium subsp. avium (MAA) were isolated from tissues of five additional adult elk. Gastrointestinal tract and lymph node tissues from 17 of the 37 elk (46%) examined had histopathologic lesions commonly seen with mycobacterial infection; however, acid-fast bacteria were not observed. All MAC infections were detected from adult elk (P = 0.023). In adult elk, a statistically significant association was found between MAA infection and ELISA sample-to-positive ratio (S/P) > or = 0.25 (P=0.021); four of five MAA culture-positive elk tested positive by ELISA. Sensitivity and specificity of ELISA S/P > or = 0.25 for detection of MAA in adult elk were 50% and 93%, respectively. No significant associations were found between MAC infection and sex or histopathologic lesions. Bacteriologic culture confirmed infection with MAP and MAA in this asymptomatic tule elk herd. The Johne's ELISA was useful in signaling mycobacterial infection on a population basis but could not discriminate between MAA and MAP antibodies. The multiplex PCR was useful in discriminating among the closely related species belonging to MAC.

MORTALITY DURING TREATMENT: FACTORS AFFECTING THE SURVIVAL OF OILED, REHABILITATED COMMON MURRES (URIA AALGE).

After major oil spills, hundreds to thousands of live stranded birds enter rehabilitative care. To target aspects of rehabilitative efforts for improvement and to evaluate which initial physical examination and biomedical parameters most effectively predict survival to release, medical records were examined from 913 Common Murres ( Uria aalge ; COMUs) oiled during the November 2001-January 2003 oil spill associated with the sunken S.S. Jacob Luckenbach off San Francisco, California, US. Results showed that 52% of all deaths occurred during the first 2 days of treatment. Birds stranding closest to the wreck had greater amounts of oil on their bodies than birds stranding farther away. More heavily oiled birds were in better clinical condition than birds with lesser amounts of oil, as shown by higher body mass (BM), packed cell volumes (PCV), total plasma protein (TP), and higher survival proportions. Additionally, BM, PCV, TP, and body temperature were positively correlated. For comparison, medical records from all nonoiled COMUs admitted for rehabilitation at the same facility during 2007-09 (n=468) were examined, and these variables were also found to be positively correlated. Oiled birds with BM under 750 g had approximately 5% lower PCV than BM-matched nonoiled COMUs. More heavily oiled COMUs may be in better condition than less oiled birds because heavily oiled birds must beach themselves immediately to avoid drowning and hypothermia, whereas lightly oiled birds may postpone beaching until exhausted due to extreme body catabolism. The strong relationship of PCV to BM regardless of oiling provides evidence that anemia commonly encountered in oiled seabirds may be a sequela to overall loss of body condition rather than solely due to toxic effects of oiling. Clinical information garnered in this study provides guidance for triage decisions during oil spills.

Ophthalmic effects of petroleum dispersant exposure on common murres (Uria aalge): An experimental study.

The safety of chemical dispersants used during oil spill responses is largely unknown in birds. We captured common murres in Monterey Bay, CA and exposed them to Corexit EC9500a, crude oil, or a combination in artificial seawater. We performed ophthalmic examinations and measured intraocular pressures and tear production before and after exposure. Loglinear analysis found that exposure to oil or dispersant was related to the development of conjunctivitis and corneal ulcers. Odds ratios for birds exposed to oil or dispersant were positive and significant for the development of conjunctivitis, while odds ratios for the development of corneal ulcers were positive and significant only for birds exposed to a high concentration of oil. Ocular exposure to dispersants and petroleum in seabirds may cause conjunctivitis and may play a role in the development of corneal ulcers. These results have implications for policymakers who develop protocols for the use of dispersants during marine oil spills.

Application of the luciferase recombinant cell culture bioassay system for the analysis of polycyclic aromatic hydrocarbons.

An aryl hydrocarbon (Ah) receptor-based luciferase cell culture bioassay developed to detect 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and other halogenated aromatics was modified and optimized to detect and quantitate polycyclic aromatics (PAHs). Twenty-four PAHs were analyzed, and subsequent EC50 and EC20 concentrations (based on the median and 20% TCDD maximal response, respectively) and appropriate induction equivalency factors (calculated by comparison to the response obtained with TCDD) were determined from dose-response experiments. Six compounds were shown to be active in the system, with benzo[k]fluoranthene > benz[a,h]anthracene indeno[1,2,3-cd]pyrene > benzo[a]pyrene > benzo[b]fluoranthene > chrysene. A complex mixture of 16 PAHs was also analyzed using this system, and overall induction equivalency (or 1-EQ) of the mixture was shown to be very similar to that predicted from the sum of the activity estimated for each individual PAH. Overall, our results strongly support the use of this system for the detection and relative quantitation of Ah receptor-active PAHs.