Gina M. Ylitalo

Influence of life-history parameters on organochlorine concentrations in free-ranging killer whales (Orcinus orca) from Prince William Sound, AK.

Certain populations of killer whales (Orcinus orca) have been extensively studied over the past 30 years, including populations that use Puget Sound, WA, the inside waters of British Columbia, Southeastern Alaska and Kenai Fjords/Prince William Sound, Alaska. Two eco-types of killer whales, transient and resident, occur in all of these regions. These eco-types are genetically distinct and differ in various aspects of morphology, vocalization patterns, diet and habitat use. Various genetic and photo-identification studies of eastern North Pacific killer whales have provided information on the male-female composition of most of these resident pods and transient groups, as well as the approximate ages, reproductive status and putative recruitment order (birth order) of the individual whales. Biopsy blubber samples of free-ranging resident and transient killer whales from the Kenai Fjords/Prince William Sound, AK region were acquired during the 1994-1999 field seasons and analyzed for selected organochlorines (OCs), including dioxin-like CB congeners and DDTs. Concentrations of OCs in transient killer whales (marine mammal-eating) were much higher than those found in resident animals (fish-eating) apparently due to differences in diets of these two killer whale eco-types. Certain life-history parameters such as sex, age and reproductive status also influenced the concentrations of OCs in the Alaskan killer whales. Reproductive female whales contained much lower levels of OCs than sexually immature whales or mature male animals in the same age class likely due to transfer of OCs from the female to her offspring during gestation and lactation. Recruitment order also influenced the concentrations of OCs in the Alaskan killer whales. In adult male residents, first-recruited whales contained much higher OC concentrations than those measured in non-first-recruited (e.g. second recruited, third recruited) resident animals in the same age group. This study provides baseline OC data for free ranging Alaskan killer whales for which there is little contaminant information.

Organochlorine contaminant exposure and associations with hematological and humoral immune functional assays with dam age as a factor in free-ranging northern fur seal pups (Callorhinus ursinus).

Potential effects of organochlorine contaminant (OC) exposure on humoral immune function and health of free-ranging northern fur seals were investigated. Forty-two neonates were captured for blood sample collection and re-sampled as pups 29-51 days later. OCs were extracted from whole blood to identify polychlorinated biphenyl congeners and chlorinated pesticides by high performance liquid chromatography. Humoral immune function was assessed by antibody responses to vaccination and immunoglobulin concentrations. Additional health status indicators included leukocyte counts and haptoglobin concentrations. Mean OC concentrations were higher in neonates than at recapture. Neonates of young dams had higher mean OC concentrations than neonates of older dams. A higher proportion of old dams pups developed a twofold or greater increase in antigen-specific antibodies compared to young dams pups. Higher OC exposure and poor immune responses in first-born pups may indicate a higher risk of secondary morbidity and mortality than pups of multiparous dams.

Screening for planar chlorobiphenyl congeners in tissues of marine biota by high-performance liquid chromatography with photodiode array detection

A rapid method has been developed to screen for planar chlorobiphenyl (CB) congeners, as well as certain other CBs and DDTs, in tissue samples from marine biota. The analytes were extracted from tissue matrices with 1:1 hexane/pentane (v/v) and interfering compounds were separated from the CBs on a gravity-flow column packed with acidic, basic and neutral silica gel eluted with 1:1 hexane/methylene chloride (v/v). Subsequently, the planar CB congeners were resolved from the DDTs and other CBs by HPLC on Cosmosil PYE analytical columns cooled to 9 degrees C and were measured by an ultraviolet (UV) photodiode array (PDA) detector. Two important advantages of PDA over conventional UV detection were the ability to identify individual analytes by comparing their UV spectra to those of reference standards and the ability to establish the spectral homogeneity (purity) of the analytes by comparing spectra within a peak to the apex spectrum. The HPLC/PDA method was tested with tissue samples from fish, shellfish and marine mammals; concentrations of certain CBs and DDTs in samples determined by screening compared favorably with those in the same samples analyzed by a comprehensive method (e.g., gas chromatography/high resolution mass spectrometry). However, the HPLC/PDA method was about an order of magnitude less sensitive than determinations by comprehensive methods.

Analyses for petroleum-related contaminants in marine fish and sediments following the Gulf oil spill

Abstract Approximately 1 year after Kuwaiti crude oil was spilled and burned during the Gulf War, scientists on board the NOAA R/V Mt Mitchell sampled sediment and biota in the Gulf to look for evidence of petroleum contamination. Subtidal sediments and fish bile were analysed for petroleum-related aromatic compounds (ACs) on board the Mt Mitchell using cost-effective and rapid high-performance liquid chromatographic (HPLC) screening methods. Parent ACs and their alkyl homologs were measured in sediments. However, because fish extensively metabolize ACs in their livers to form polar metabolites that are concentrated in bile for excretion, metabolites of ACs were measured in bile. HPLC screening analyses found ACs in many of the sediment and fish bile samples collected. The results of HPLC screening were confirmed in selected samples by gas chromatography/mass spectrometry (GC/MS) analyses that identified petroleum-related ACs (e.g., alkyl phenanthrenes or dibenzothiophenes) in sediments and their metabolites in bile.

The role of domoic acid in abortion and premature parturition of California sea lions (Zalophus californianus) on San Miguel Island, California.

Domoic acid is a glutaminergic neurotoxin produced by marine algae such as Pseudo-nitzschia australis. California sea lions (Zalophus californianus) ingest the toxin when foraging on planktivorous fish. Adult females comprise 60% of stranded animals admitted for rehabilitation due to acute domoic acid toxicosis and commonly suffer from reproductive failure, including abortions and premature live births. Domoic acid has been shown to cross the placenta exposing the fetus to the toxin. To determine whether domoic acid was playing a role in reproductive failure in sea lion rookeries, 67 aborted and live-born premature pups were sampled on San Miguel Island in 2005 and 2006 to investigate the causes for reproductive failure. Analyses included domoic acid, contaminant and infectious disease testing, and histologic examination. Pseudo-nitzschia spp. were present both in the environment and in sea lion feces, and domoic acid was detected in the sea lion feces and in 17% of pup samples tested. Histopathologic findings included systemic and localized inflammation and bacterial infections of amniotic origin, placental abruption, and brain edema. The primary lesion in five animals with measurable domoic acid concentrations was brain edema, a common finding and, in some cases, the only lesion observed in aborted premature pups born to domoic acid–intoxicated females in rehabilitation. Blubber organochlorine concentrations were lower than those measured previously in premature sea lion pups collected in the 1970s. While the etiology of abortion and premature parturition was varied in this study, these results suggest that domoic acid contributes to reproductive failure on California sea lion rookeries.

Federal seafood safety response to the Deepwater Horizon oil spill.

Following the 2010 Deepwater Horizon oil spill, petroleum-related compounds and chemical dispersants were detected in the waters of the Gulf of Mexico. As a result, there was concern about the risk to human health through consumption of contaminated seafood in the region. Federal and Gulf Coast State agencies worked together on a sampling plan and analytical protocols to determine whether seafood was safe to eat and acceptable for sale in the marketplace. Sensory and chemical methods were used to measure polycyclic aromatic hydrocarbons (PAHs) and dispersant in >8,000 seafood specimens collected in federal waters of the Gulf. Overall, individual PAHs and the dispersant component dioctyl sodium sulfosuccinate were found in low concentrations or below the limits of quantitation. When detected, the concentrations were at least two orders of magnitude lower than the level of concern for human health risk. Once an area closed to fishing was free of visibly floating oil and all sensory and chemical results for the seafood species within an area met the criteria for reopening, that area was eligible to be reopened. On April 19, 2011 the area around the wellhead was the last area in federal waters to be reopened nearly 1 y after the spill began. However, as of November 9, 2011, some state waters off the Louisiana coast (Barataria Bay and the Delta region) remain closed to fishing.

Transplacental transfer of organochlorines in california sea lions (Zalophus californianus)

The transplacental transfer of organochlorines (OCs) in California sea lions (Zalophus californianus) was investigated by analyzing blubber samples from 20 female sea lions and their fetuses during the last trimester of pregnancy. A rapid, high-performance liquid chromatographic, photodiode-array method was used to measure blubber concentrations of polychlorinated biphenyls (PCBs), including dioxin-like congeners, as well as DDTs and hexachlorobenzene. Summed values of PCBs (sigmaPCBs), of DDTs (sigmaDDTs), and of PCB toxic equivalents (sigmaPCB TEQs) were calculated from these data. The ratios of mean blubber concentrations of fetal sigmaPCBs to maternal blubber concentrations of sigmaPCBs were 0.45 by wet weight and 0.97 by lipid weight, but these ratios varied widely among mother-fetus pairs. Mean ratios of fetal sigmaDDTs to maternal sigmaDDTs were 0.53 by wet weight and 1.12 by lipid weight. Fetuses were classified into two age groups, based on date of recovery, to examine differences in OC transfer because of gestational age. Fetal to maternal ratios for individual PCB congeners, DDT compounds, and sigmaPCBs, sigmaDDTs, and sigmaPCB TEQs were lower among premature compared with late-term fetuses. These ratios increased for both groups as the logarithmic n-octanol/water partition coefficient (log Kow) for each compound decreased. Linear predictions for sigmaPCB and sigmaDDT concentrations in fetal blubber could be obtained using the sigmaPCB and sigmaDDT concentrations in maternal blubber, maternal and fetal blubber lipid content, maternal mass, and maternal age. Fetal TEQ was explained by maternal TEQ and maternal age. The ability to predict contaminant concentrations in fetal blubber from maternal parameters is important for developing risk assessment models for marine mammals.

Rapid high-performance liquid chromatographic methods that screen for aromatic compounds in environmental samples

Abstract In order to monitor the quality of coastal waters that provide habitats for living marine resources, samples of sediment and biota must be analyzed to assess the degree and distribution of anthropogenic contamination. Analytical time and costs can be greatly reduced by first employing methods that screen for contaminants before selecting samples for rigorous analyses. In this paper, we review the applications of rapid high-performance liquid chromatographic (HPLC) methods to screen for aromatic compounds in sediment, bile and tissue samples. These methods have been used to assess damage to natural resources after the Exxon Valdez oil spill. In addition, the bile screening method has also been used to evaluate contaminants in fish sampled for a national monitoring program. The rapid screening of sediment or bile provides an estimate of contaminant concentrations that can then be confirmed in selected samples by more complicated and expensive analyses by gas chromatography/3-mass spectrometry (GC-MS). Furthermore, HPLC and GC-MS chromatographic patterns from sediment and bile can provide information about the source of contamination, e.g., crude oil, diesel fuel or pyrogenic contaminants. We also discuss the important role screening methods will play in the future in assessing the quality of aquatic habitats, the safety of seafood, and other important issues related to anthropogenic contamination.

Rapid, semi-quantitative screening of sediments for aromatic compounds using sonic extraction and HPLC/fluorescence analysis☆

Abstract Awareness of contamination from anthropogenic sources in the environment has led to a demand for increasing numbers of analyses to evaluate the extent of the problem. As a result, there is a need for rapid, low cost methods to screen for contaminants in environmental samples to allow priorities to be set for the more expensive, detailed analyses. In this paper, we report a method to screen sediments for aromatic hydrocarbons. The method uses sonic extraction followed by size-exclusion HPLC with fluorescence detection to estimate the concentrations of fluorescent aromatic compounds in sediment samples. The concentrations of aromatic compounds determined by the screening method are highly correlated with the sum of the aromatic hydrocarbons determined by a standard analytical method which uses HPLC cleanup and GC/MS quantitation.

Blood-based biomarkers of selenium and thyroid status indicate possible adverse biological effects of mercury and polychlorinated biphenyls in Southern Beaufort Sea polar bears.

We examined biomarkers of selenium status (whole blood Se; serum Se; glutathione peroxidase activity) and thyroid status (concentrations and ratios of thyroxine, T4; tri-iodothyronine, T3; albumin) in polar bears to assess variations among cohorts, and relationships to circulating concentrations of contaminants. Concentrations of total mercury (Hg) in whole blood were similar among cohorts (prime aged males and females, older animals, ages≥16 years, and young animals, ages 1-5 years; 48.44±35. 81; p=0.253). Concentrations of sum of seven polychlorinated biphenyls (∑PCB7) in whole blood were greater in females (with and without cubs, 26.44±25.82 ng/g ww) and young (26.81±10.67 ng/g ww) compared to males (8.88±5.76 ng/g ww, p<0.001), and significantly related to reduced body condition scores (p<0.001). Concentrations of Se and albumin were significantly greater in males than females (whole blood Se, males, 42.34 pmol/g ww, females, 36.25±6.27 pmol/g ww, p=0.019; albumin, males, 4.34±0.34 g/dl, females, 4.10±0.29 g/dL, p=0.018). Glutathione peroxidase activity ranged from 109.1 to 207.8 mU/mg hemoglobin, but did not differ significantly by sex or age (p>0.08). Thyroid hormones were greater in females (solitary females and females with cubs) compared to males (p<0.001). Biomarkers of Se status and concentrations of T3 were significantly positively related to Hg in all prime aged polar bears (p<0.03). Albumin concentrations were significantly positively related to total TT4, and significantly negatively related to concentrations of ∑PCB7 (p<0.003). Total thyroxine (TT4) was significantly negatively associated with blood concentrations of ∑PCB7 in solitary females (p=0.045). These data suggest that female polar bears were more susceptible to changes in blood-based biomarkers of selenium and thyroid status than males. Further classifications of the physiologic states of polar bears and repeated measures of individuals over time are needed to accurately assess the biological impact of combined toxicant exposures.