Brenda E. Ballachey

Cytochrome P4501A biomarker indication of oil exposure in harlequin ducks up to 20 years after the Exxon Valdez oil spill.

Hydrocarbon-inducible cytochrome P4501A (CYP1A) expression was measured, as ethoxyresorufin-O-deethylase (EROD) activity, in livers of wintering harlequin ducks (Histrionicus histrionicus) captured in areas of Prince William Sound, Alaska, USA, oiled by the 1989 Exxon Valdez spill and in birds from nearby unoiled areas, during 2005 to 2009 (up to 20 years following the spill). The present work repeated studies conducted in 1998 that demonstrated that in harlequin ducks using areas that received Exxon Valdez oil, EROD activity was elevated nearly a decade after the spill. The present findings strongly supported the conclusion that average levels of hepatic EROD activity were higher in ducks from oiled areas than those from unoiled areas during 2005 to 2009. This result was consistent across four sampling periods; furthermore, results generated from two independent laboratories using paired liver samples from one of the sampling periods were similar. The EROD activity did not vary in relation to age, sex, or body mass of individuals, nor did it vary strongly by season in birds collected early and late in the winter of 2006 to 2007, indicating that these factors did not confound inferences about observed differences between oiled and unoiled areas. We interpret these results to indicate that harlequin ducks continued to be exposed to residual Exxon Valdez oil up to 20 years after the original spill. This adds to a growing body of literature suggesting that oil spills have the potential to affect wildlife for much longer time frames than previously assumed.

Mitochondrial-DNA variation among subspecies and populations of sea otters (Enhydra lutris)

We used restriction-enzyme analysis of polymerase-chain reaction-amplified, mitochondrial DNA (mtDNA) to assess genetic differentiation of subspecies and populations of sea otters, Enhydra lutris , throughout the range of the species. There were several haplotypes of mtDNA in each subspecies and geographically separate populations. MtDNA sequence divergence of haplotypes of sea otters was 0.0004–0.0041 base substitutions per nucleotide. E. L nereis appears to have monophyletic mitochondrial DNA, while E. I. lutris and E. I. kenyoni do not. Different frequencies of haplotypes of mtDNA among populations reflect current restriction of gene flow and the unique histories of different populations. There are two or three haplotypes of mtDNA and diversity of haplotypes is 0.1376–0.5854 in each population of otters. This is consistent with theoretical work, which suggests that population bottlenecks of sea otters probably did not result in major losses of genetic variation for individual populations, or the species as a whole.

Could residual oil from the Exxon Valdez spill create a long-term population ''sink'' for sea otters in Alaska?

Over 20 years ago, the Exxon Valdez oil tanker spilled 42 million L of crude oil into the waters of Prince William Sound, Alaska, USA. At the time of the spill, the sea otter (Enhydra lutris) population inhabiting the spill area suffered substantial acute injuries and loss. Subsequent research has resulted in one of the best-studied species responses to an oil spill in history. However, the question remains: Is the spill still influencing the Prince William Sound sea otter population? Here we fit time-varying population models to data for the sea otter population of western Prince William Sound to quantify the duration and extent of mortality effects from the spill. We hypothesize that the patchy nature of residual oil left in the environment has created a source-sink population dynamic. We fit models using the age distributions of both living and dying animals and estimates of sea otter population size to predict the number of sea otters in the hypothesized sink population and the number lost to this sink due to chronic exposure to residual oil. Our results suggest that the sink population has remained at just over 900 individuals (95% CI: 606-960) between 1990 and 2009, during which time prime-age survival remained 2-6% below pre-spill levels. This reduced survival led to chronic losses of ;900 animals over the past two decades, which is similar in magnitude to the number of sea otter deaths documented in western Prince William Sound during the acute phase of the spill. However, the unaffected source population appears to be counterbalancing these losses, with the model indicating that the sea otter population increased from ;2150 individuals in 1990 to nearly 3000 in 2009. The most optimistic interpretation of our results suggests that mortality effects dissipated between 2005 and 2007. Our results suggest that residual oil can affect wildlife populations on time scales much longer than previously believed and that cumulative chronic effects can be as significant as acute effects. Further, source-sink population dynamics can explain the slow recovery observed in the spill-affected western Prince William Sound sea otter population and are consistent with available data.

CHEMICAL ANESTHESIA OF NORTHERN SEA OTTERS (ENHYDRA LUTRIS): RESULTS OF PAST FIELD STUDIES

Abstract Between 1987 and 1997, we chemically immobilized 597 wild sea otters (Enhydra lutris) in Alaska for the collection of biological samples or for surgical instrumentation. One drug-related sea otter fatality occurred during this time. Fentanyl in combination with diazepam produced consistent, smooth inductions with minimal need for supplemental anesthetics during procedures lasting 30–40 min. Antagonism with naltrexone or naloxone was rapid and complete, although we observed narcotic recycling in sea otters treated with naloxone. For surgical procedures, we recommend a fentanyl target dose of 0.33 mg/kg of body mass and diazepam at 0.11 mg/kg. For nonsurgical biological sample collection procedures, we recommend fentanyl at 0.22 mg/kg and diazepam at 0.07 mg/kg. We advise the use of the opioid antagonist naltrexone at a ratio of 2:1 to the total fentanyl administered during processing.

Gene transcription in sea otters (Enhydra lutris); development of a diagnostic tool for sea otter and ecosystem health

Gene transcription analysis for diagnosing or monitoring wildlife health requires the ability to distinguish pathophysiological change from natural variation. Herein, we describe methodology for the development of quantitative real‐time polymerase chain reaction (qPCR) assays to measure differential transcript levels of multiple immune function genes in the sea otter (Enhydra lutris); sea otter‐specific qPCR primer sequences for the genes of interest are defined. We establish a ‘reference’ range of transcripts for each gene in a group of clinically healthy captive and free‐ranging sea otters. The 10 genes of interest represent multiple physiological systems that play a role in immuno‐modulation, inflammation, cell protection, tumour suppression, cellular stress response, xenobiotic metabolizing enzymes, antioxidant enzymes and cell–cell adhesion. The cycle threshold (CT) measures for most genes were normally distributed; the complement cytolysis inhibitor was the exception. The relative enumeration of multiple gene transcripts in simple peripheral blood samples expands the diagnostic capability currently available to assess the health of sea otters in situ and provides a better understanding of the state of their environment.

Cytochrome P4501A biomarker indication of the timeline of chronic exposure of Barrow's goldeneyes to residual Exxon Valdez oil.

We examined hepatic EROD activity, as an indicator of CYP1A induction, in Barrows goldeneyes captured in areas oiled during the 1989 Exxon Valdez spill and those from nearby unoiled areas. We found that average EROD activity differed between areas during 2005, although the magnitude of the difference was reduced relative to a previous study from 1996/1997, and we found that areas did not differ by 2009. Similarly, we found that the proportion of individuals captured from oiled areas with elevated EROD activity (≥ 2 times unoiled average) declined from 41% in winter 1996/1997 to 10% in 2005 and 15% in 2009. This work adds to a body of literature describing the timelines over which vertebrates were exposed to residual Exxon Valdez oil and indicates that, for Barrows goldeneyes in Prince William Sound, exposure persisted for many years with evidence of substantially reduced exposure by 2 decades after the spill.

Challenges to sea otter recovery and conservation

Similar to other species that in recent centuries experienced unregulated human exploitation, sea otters were extirpated throughout large portions of their historic range in the North Pacific. For most of the twentieth century, with cessation of the fur trade and because of concerted efforts at conservation, sea otters recovered much of their historic range and abundance. Late in the twentieth century, increased predation by killer whales in southwest Alaska drove sea otter populations to a few percentage points of their prior abundance, and one of the nation’s largest oil spills in south-central Alaska caused the death of several thousand animals and required more than two decades for recovery. In California, entanglement in fishing gear and environmental degradation, among other factors, have contributed to slow growth in sea otter abundance. We discuss the role of density dependence and spatial structuring of populations in reduced rates of sea otter recovery recently detected in the Northeast Pacific, and consider the potential effects of multiple low-level and cumulative threats on sea otter populations. The resilience demonstrated by sea otters over the past century will be tested in upcoming decades as human activities continue to degrade nearshore coastal areas of the North Pacific.

Cessation of oil exposure in harlequin ducks after the Exxon Valdez oil spill: Cytochrome P4501A biomarker evidence

The authors quantified hepatic hydrocarbon-inducible cytochrome P4501A (CYP1A) expression, as ethoxyresorufin-O-deethylase (EROD) activity, in wintering harlequin ducks (Histrionicus histrionicus) captured in Prince William Sound, Alaska (USA), during 2011, 2013, and 2014 (22-25 yr following the 1989 Exxon Valdez oil spill). Average EROD activity was compared between birds from areas oiled by the spill and those from nearby unoiled areas. The present study replicated studies conducted from 1998 to 2009 demonstrating that harlequin ducks using areas oiled in 1989 had elevated EROD activity, indicative of oil exposure, up to 2 decades post spill. In the present study, it was found that average EROD activity during March 2011 was significantly higher in wintering harlequin ducks captured in oiled areas relative to unoiled areas, which the authors interpret to indicate that harlequin ducks continued to be exposed to residual Exxon Valdez oil up to 22 yr after the original spill. However, the 2011 results also indicated reductions in exposure relative to previous years. Average EROD activity in birds from oiled areas was approximately 2 times that in birds from unoiled areas in 2011, compared with observations from 2005 to 2009, in which EROD activity was 3 to 5 times higher in oiled areas. It was also found that average EROD activity during March 2013 and March 2014 was not elevated in wintering harlequin ducks from oiled areas. The authors interpret these findings to indicate that exposure of harlequin ducks to residual Exxon Valdez oil abated within 24 yr after the original spill. The present study finalizes a timeline of exposure, extending over 2 decades, for a bird species thought to be particularly vulnerable to oil contamination in marine environments. Environ Toxicol Chem 2017;36:1294-1300. Published 2016 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

Implementation of Biomarker-Based Studies

Biomarkers may be used to determine chemical exposure and effects in several important ways. These include (1) evaluation of new agricultural or industrial chemicals for effects, (2) screening of municipal or industrial effluents, (3) determining the geographic distribution of chemical effects in the environment and their changes over time, (4) determining the identity and source of chemical pollutants, and (5) establishing cause and effect linkages.