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Contaminants and Biomarker Assays in Beluga Whales from the Eastern Chukchi Sea

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Contaminants and Biomarker Assays in Beluga Whales from the Eastern Chukchi Sea CONTAMINANTS AND BIOMARKER ASSAYS IN BELUGA WHALES FROM THE EASTERN CHUKCHI SEA NORTH SLOPE BOROUGH DEPARTMENT OF WILDLIFE MANAGEMENT PROGRESS REPORT, CONTRACT 2013-017 Submitted to: Dr. Robert Suydam North Slope Borough Department of Wildlife Management Barrow, Alaska 99723 (907) 852-0350 Submitted by: Mote Marine Laboratory Directorate of Marine Biology & Conservation 1600 Ken Thompson Parkway Sarasota, FL 34236 (941) 388-4441 (941) 388-4223 FAX Dr. Dana L. Wetzel and Dr. John E. Reynolds Mote Marine Laboratory, 1600 Ken Thompson Pkwy, Sarasota, FL 34236 2 March 2015 Mote Marine Laboratory Technical Report Number 1880 PROGRESS REPORT SUBMITTED TO THE NORTH SLOPE BOROUGH, DEPARTMENT OF WILDLIFE MANAGEMENT PROJECT TITLE: Contaminants and Biomarker Assays in Beluga Whales from the Eastern Chukchi Sea CONTRACT NUMBER: 2013-017 DATE: 2 March 2015 SUBMITTED TO: Dr. Robert Suydam SUBMITTED BY: Dr. Dana Wetzel and Dr. John Reynolds Mote Marine Laboratory, Sarasota, Florida PROJECT END DATE: The anticipated end date is December 2015. At that time all funds will be spent and all analyses will be completed and interpreted. A final report will be submitted by 1 January 2016. Introduction: The project has three primary components, namely 1) age determination, 2) analysis of polycyclic aromatic hydrocarbons and other classes of organic contaminants, and (3) assessment of selected biomarkers of effects of stressors. The results achieved to date will be described separately in this report. Samples (eye lens, serum, blubber, and liver) were acquired by a team of scientists led by Dr. Robert Suydam (North Slope Borough, Department of Wildlife Management; NSB) during the 2012 subsistence harvest of beluga whales (Delphinapterus leucas) by the community of Point Lay, Alaska. The list of samples collected for age determination and biomarker analyses, together with morphometric and other data for each whale appears in Table 1. In addition, samples of liver and blubber were collected and sent to Mote Marine Laboratory for the contaminant analyses. The overarching goals of the project are to (1) assess the results of a range of analyses (to assess beluga whale age, contaminant levels, and sublethal biomarkers of effects of stressors) in association with other data (e.g., morphology, body condition) to better understand the status of the Chukchi Sea beluga whale stock, and (2) create “baseline” values against which to assess changes in beluga whale status in the future. 1 Age determination: Background and Goals: The goals of this component of the project were as follows: ---to use amino acid racemization (aar) techniques to assess the age of individual beluga whales harvested for subsistence in Point Lay, Alaska; and ---to compare ages determined using aar with ages determined using counts of growth layer groups in the teeth of the same individual whales. Suydam sent frozen eyes from 15 whales (Table 1) harvested prior to the involvement of Wetzel/Reynolds in the Point Lay hunt, or from whales harvested elsewhere, to Mote Marine Laboratory for analysis. This sample included one fetus (LDL-1F-09). Under the current contract, the eyes from an additional 8 whales were taken for subsistence were collected and frozen for later shipment to and analysis at Mote Marine Laboratory (Table 1). Eyes of individual belugas were processed and analyses conducted as described by Wetzel et al. (2014; Appendix 1) for bowhead whales (Balaena mysticetus). The analyses include artificial aging studies of samples from the lens of one of the beluga whales sampled. Wetzel et al. (2014) is provided (Appendix 1) in its entirety. The chemical analyses of the hydrolyzed and derivative eye lens nucleus of each whale yield values for the D form of aspartic acid and the L form of aspartic acid (D/L). Kasp represents the rate constant for the conversion of the L form to the D form in the living animal; that rate is temperature dependent, so the calculated age for each whale is also temperature related. The analyses provide the necessary data to apply the following equation to calculate age of each whale: log (1 (DL /LD )ii ) / (1 ( / ) )log(1(D/L) 00)/ (1 (/D L) ) Agei 2Kasp Kasp is calculated using the results of the heating experiments (Wetzel et al. 2014) using the Arrhenius equation: KEasp Aexp a / (RT ) Here, R is the universal gas constant, and T is the temperature of the living whale in degrees Kelvin. The parameters A and Ea are estimable from the heating experiment data. 2 Results: As noted, lens temperature in the living whale has a significant influence on the calculated age of the whale. To provide a better understanding of lens temperature in living belugas, Hans Thewissen and Andrew VonDuyke (members of the Point Lay scientific group) used a probe and thermistor to determine the lens temperature within two hours after death of six whales harvested in summer, 2013. Those temperatures (Thewissen, pers. comm. 2013; Table 2) averaged 18.57°C. In 2012, Thewissen (pers. comm. 2012) acquired lens temperatures from three harvested whales that had been dead between 50 and 80 minutes; for those whales the lens temperatures were 23.7°C, 18.8°C, and 20.1°C (mean = 20.866°C, a figure that corresponds well with the average temperature [=20.85°C] of the two whales sampled most quickly after death in 2013; Table 2). Note that these measurements provide usual insight and confirm that beluga lenses are colder than is core body temperature, but given circumstance of the measurements (i.e., the relatively high temperature of the ocular fat, and the time interval warming between death and the temperature measurements), it seems quite possible that lens temperature in the living whales is colder. Table 3 provides replicate data for the measured D form and L form of aspartic acid for each beluga whales sampled in 2012 for this study (i.e., individuals with a field number of LDL##12), as well as additional individuals for which eyes were archived. The measurements are extremely precise, as demonstrated by the very small standard deviations. In Table 3, several values of Kasp are provided at plausible lens temperatures to provide the calculated age of each whale. Note that one column of possible ages used the derived value of Kasp based on the empirical measurements taken in 2012 of lens temperature described above (i.e., mean = 20.866°C). Discussion and Implications: Although beluga eyes appear not to be as cold as those of bowhead whales (estimated to be around 11.3°C or lower based on some direct measurements following harvest; see Wetzel et al. 2014; Sformo et al. 2011), it appears that the beluga eyes are likely to be considerably cooler (perhaps around 15°C or less) than the “normal” mammalian body temperature and beluga whale core temperature. The whales from the 2012 harvest for which age was derived (Table 3) were generally young animals, with only two exceeding 10 years of age based on calculations using a lens temperature of 15°C. Sexual maturity in beluga whales occurs at age 9-12 years in females and sometime later in males (O’Corry-Crowe 2009). The animals turn white in color a couple years earlier than that in both sexes, and the ages of the larger, white whales sampled in 2012 approach or equal the age at which white pigmentation predominates. In K-selected species, mature adult females constitute the most valuable demographic group to sustain population size, so if the whales we examined are representative of those taken for subsistence along the northwestern coast of Alaska, it appears that the hunt may largely be targeting immature animals that travel together. This suggests that the harvest may avoid 3 targeting the most valuable demographic group of belugas, an important management consideration. There are several factors that enter into accurate calculations of age using D:L ratios. One is the D:L ratio in the lens nucleus of a whale at age 0. In this study we had access to an eye from only one fetal beluga; additional fetal eyes would be valuable in calculating D:L for whales at age 0. A compelling reason for doing this study is to compare ages derived from amino acid racemization methods (described here) with ages derived from counting growth layer groups (GLGs) in the teeth of belugas. The latter approach requires validation for beluga whales for which there are uncertainties regarding deposition rates of GLGs and loss of GLGs with age (e.g., Lockyer et al. 2007). The NSB-DWM hoped that this study would help to resolve the questions about the utility of GLGs. The comparative GLG data for the whales we examined will be provided by Suydam, but are not yet available. Next Steps: The obvious next steps for the age determination component of the project include (a) acquiring age estimates based on GLG analysis, (b) comparing the GLG-based ages with the aar-based ages, and (c) developing a manuscript for a peer-reviewed journal. However, two specific steps that would be useful for calculating ages based on the aar approach include: (a) acquiring frozen eyes of at least two additional fetuses so better document the D:L ratio at age 0, and (b) acquiring, if possible a frozen eye from Naku, a known age adult female whale that recently died at Mystic, along with temperature of the water in her tank. Contaminant Analyses: Background and Goals: The effects of contaminants are a major concern in the Arctic from the standpoints of both human health and wildlife health. Given the current and past extent of oil and gas exploration and development, it is extremely important to assess levels of PAHs (polycyclic aromatic hydrocarbons), which include the most toxic components of oil.
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