National Biomonitoring Specimen Bank Research Articles

The International Society for Biological and Environmental Repositories Presents Abstracts from Its Annual Meeting Due North: Aligning Biobanking Practice with Evolving Evidence and Innovation May 9–12, 2017 Toronto, Canada

The International Society for Biological and Environmental Repositories Presents Abstracts from Its Annual Meeting Due North: Aligning Biobanking Practice with Evolving Evidence and Innovation May 9–12, 2017 Toronto, Canada

Design and Applications of the National Institute of Standards and Technology's (NIST's) Environmental Specimen Banking Programs

The National Institute of Standards and Technology (NIST) environmental specimen banking system consists of two environmental specimen banks (ESBs): the National Biomonitoring Specimen Bank established in 1979, and the Marine Environmental Specimen Bank established in 2002. Both facilities were specifically designed to store environmental specimens over long periods of time (50–100 years) and in such a way that future researchers could use these specimens to answer questions regarding trends in newly recognized environmental contaminants and verification of past analytical results. The NIST environmental banking system maintains collections of human liver specimens, human blood serum and blood spots, human diet samples, marine sediments, fish tissues, mussels, oysters, marine mammal tissues, and bird eggs and feathers collected as part of several monitoring and research programs supported by the U.S. Government. The NIST environmental banking system emphasizes: (1) carefully designed (and published) collection and banking procedures, (2) cryogenic storage to ensure sample stability, (3) high efficiency particulate air (HEPA)-filtered clean air conditions in the sample preparation and freezer rooms, (4) cryogenic homogenization systems for sample preparation, (5) computerized sample inventory and tracking system, (6) computerized security and monitoring systems, and (7) redundancy to minimize sample loss due to equipment or system failure. The NIST ESBs provide a resource of research specimens that are used to address questions regarding temporal and geographic trends in environmental contamination (including documentation of newly recognized contaminants), changes in ecosystem structure and function, genetic separation of populations of animals, and the health status of marine animals.

The U.S. National Biomonitoring Specimen Bank and the Marine Environmental Specimen Bank

The National Biomonitoring Specimen Bank (NBSB), established in 1979 at the NIST Neutron Research Facility, Gaithersburg, Maryland, was specifically designed to store environmental specimens over long periods of time (50-100 years). This bank contains specimens (e.g., human livers, marine sediments, fish tissues, mussels, oysters, human diet samples, and marine mammal tissues) collected as part of several monitoring and research programs supported by U.S. Federal agencies. In 2002, NIST completed the construction of a second environmental specimen bank facility specifically designed for supporting monitoring and research on marine environmental health issues. This facility, the Marine Environmental Specimen Bank (Marine ESB) is located at the Hollings Marine Laboratory in Charleston, South Carolina, in partnership with a U.S. Federal resource agency, two universities, and a State of South Carolina resource agency. The Marine ESB provides a resource of research specimens that are used to address questions regarding temporal and geographic trends in environmental contamination, genetic separation of populations of animals, and the health status of various types of marine animals. Specimens banked include marine mammal tissues, bird tissues, mussels, and oysters. Plans are underway to establish protocols and initiate banking procedures for other types of marine organisms and environmental materials as part of an expanded effort to support research on the health of marine biota.

Persistent organic pollutants in Alaskan ringed seal (Phoca hispida) and walrus (Odobenus rosmarus) blubber

Since 1987, the Alaska Marine Mammal Tissue Archival Project (AMMTAP) has collected tissues from 18 marine mammal species. Specimens are archived in the National Institute of Standards and Technology's National Biomonitoring Specimen Bank (NIST-NBSB). AMMTAP has collected blubber, liver and/or kidney specimens from a number of ringed seals (Phoca hispida) from the areas near Nome and Barrow, Alaska and walruses (Odobenus rosmarus) from several locations in the Bering Sea. Thirty-three ringed seal and 15 walrus blubber samples from the NIST-NBSB were analyzed for persistent organic pollutants (POPs). The compounds determined included PCBs (28 congeners or congener groups), DDT and related compounds, hexachlorobenzene (HCB), hexachlorocyclohexane isomers (HCHs), chlordanes, dieldrin, and mirex. POP concentrations in ringed seal blubber were significantly higher in Barrow than in Nome when statistically accounting for the interaction of age and gender; HCB, however, was not statistically different between the two locations. Unlike males, POP concentrations and age were not significantly correlated in females probably as a result of lactational loss. POP concentrations in walrus blubber were lower than in ringed seal blubber for SigmaPCBs, chlordanes, and HCHs, but higher for dieldrin and mirex. POP concentrations in ringed seals and walrus from Alaska provide further evidence that the western Arctic tends to have lower or similar POP concentrations compared to the eastern Canadian Arctic.

Persistent organochlorine pollutants in ringed seals and polar bears collected from northern Alaska

Blubber samples from ringed seal (Phoca hispida; n = 8) and polar bear subcutaneous fat (Ursus maritimus; n = 5) were collected near Barrow, Alaska in 1996 as part of the Alaska Marine Mammal Tissue Archival Project (AMMTAP) and retained in the National Biomonitoring Specimen Bank at the National Institute of Standards and Technology in Gaithersburg, Maryland (USA). The samples were analyzed for a variety of persistent organochlorine pollutants (POPs) including polychlorinated biphenyls (PCBs), hexachlorocyclohexanes (HCHs), chlordane and metabolites, hexachlorobenzene (HCB) and DDTs and metabolites. The geometric mean, on a wet mass basis, of sigmaPCBs (sum of 29 congeners and congener groups) were 732+/-282 ng/g (1 S.D.) in seals and 3395+/-1442 ng/g in polar bears. The geometric mean of sigmaDDTs, sigmaHCHs (alpha-, beta- and gamma- HCH) and HCB concentrations (wet mass basis) in seals and bears were 562+/-261 ng/g vs. 74.8+/-39 ng/g, 380+/-213 ng/g vs. 515 ng/g, and 17.4+/-10.1 ng/g vs. 183+/-153 ng/g, respectively. The geometric mean sum of chlordane (sigmachlordane, sum of cis- and trans-chlordane, cis- and trans-nonachlor, oxychlordane and heptachlor epoxide) and dieldrin concentrations in ringed seals and polar bears were 753+/-617 ng/g vs. 720+/-315 ng/g and 38.6+/-22.8 ng/g vs. 130+/-65 ng/g, respectively. Apparent bioaccumulation factors (polar bear/ringed seal POP concentrations) were lower in the animals sampled near Barrow, Alaska than in those from locations in the Canadian Arctic. This suggests that polar bears are also preying on marine mammals from lower trophic levels than the ringed seals with correspondingly lower organochlorine levels, such as bowhead whale carcasses. PCB congener patterns in the samples demonstrated the metabolism of certain PCB congeners in the polar bear relative to the ringed seal in agreement with previous studies. Regional comparisons of animals collected in Alaska and Arctic Canada are presented.

Quality assurance in analysis of cryogenically stored liver tissue specimens from the NIST National Biomonitoring Specimen Bank (NBSB)

Portions of liver tissue specimens originally stored in the National Biomonitoring Specimen Bank (NBSB) and analyzed between 1980 and 1987 were re-analyzed in 1997 using instrumental neutron activation analysis (INAA) for the determination of 17 trace elements. Duplicate portions of each specimen had been stored at two different temperatures. The first was stored in a liquid nitrogen vapor-cooled freezer at −150°C, standard NBSB storage conditions, and the other in an electric freezer maintained at −80°C. Two portions of seven livers from each storage temperature were re-analyzed for this work. Results showed no changes in trace element content as a function of storage temperature, within the uncertainty of the method used. Results from these analyses agreed with results of initial analyses for most analytes in most sub-samples. Of the exceptions, five were due, in part, to an incorrect basis mass for the initial sub-specimen of one tissue, five with variable Zn results were attributed to difficulties in peak fitting for this element during INAA data processing, and the remaining were isolated differences discussed in this paper. Results of this work indicate that specimen storage and processing protocols are adequate to prevent noticeable contamination of specimens with trace elements, with the exception of Cr. Variability in Cr content was observed for the liver tissues which may have been caused by Cr contamination of the samples by the Teflon mill. Analyses of portions of Standard Reference Material (SRM) 1566a Oyster Tissue (certified in 1989) and SRM 1577a Bovine Liver (certified in 1982) were also included in this study for the purpose of quality control and to assess the stability of these freeze-dried powders that were stored at room temperature. No changes were observed in these materials.

Concentrations of chlorinated hydrocarbons and trace elements in marine mammal tissues archived in the U.S. national biomonitoring specimen bank

The U.S. National Biomonitoring Specimen Bank (NBSB) provides for the long term storage of well documented and preserved specimens representing several types of environmental matrices. A major part of this inventory consists of marine mammal tissues (e.g., blubber, liver, kidney, and muscle). Within the NBSB selected specimens are periodically analyzed for chlorinated hydrocarbons and trace elements. Although only 20% of the 560 marine mammal specimens in the NBSB have been analyzed, the database is of value in evaluating the stability of analytes and sample degradation during storage, for comparing with results from samples collected in the future for long-term monitoring, and for comparing with analytical results from other laboratories on samples collected at the same time for monitoring purposes. The NBSB analytical database contains results for 37 elements, many of which are not analyzed routinely by conventional analytical techniques used in monitoring programs, and the following organic compounds: selected PCB congeners, DDT compounds, α- and γ-HCH, HCB, heptachlor epoxide, oxychlordane, cis-chlordane, trans-chlordane, cis-nonachlor, trans-nonachlor, and dieldrin in 9 marine mammal species: northern fur seal ( Callorhinus ursinus), ringed seal ( Phoca hispida), spotted seal ( P. largha), bearded seal ( Erignathus barbatus), pilot whale ( Globicephala melas), harbor porpoise ( Phocoena phocoena), white-sided dolphin ( Lagenorhynchus acutus), beluga whale ( Delphinapterus leucas), and bowhead whale ( Balaena mysticetus). Analyses of beluga whale blubber for toxaphene and additional chlorinated hydrocarbons are obtained through collaboration with the Department of Fisheries and Oceans Canada.

Specimen banking of marine organisms in the United States: Current status and long-term prospective

A major part of the activities conducted over the last decade by the National Biomonitoring Specimen Bank (NBSB) has involved the archival of marine specimens collected by ongoing environmental monitoring programs. These archived specimens include bivalves, marine sediments, and fish tissues collected by the National Status and Trends and the Exxon Valdez Oil Spill Damage Assessment programs, and marine mammal tissues collected by the Marine Mammal Health and Stranding Response Program and the Alaska Marine Mammal Tissue Archival Project. In addition to supporting these programs, the specimens have been used to investigate circumpolar patterns of chlorinated hydrocarbon concentrations, genetic separation of marine animal stocks, baseline levels of essential and nonessential elements in marine mammals, and the potential risk to human consumers in the Arctic from anthropogenic contaminants found in local subsistence foods. The NBSB specimens represent a resource that has the potential for addressing future issues of marine environmental quality and ecosystem changes through retrospective analysis; however, an ecosystem-based food web approach would maximize this potential. The current status of the NBSB activities related to the banking of marine organisms is presented and discussed, the long-term prospective of these activities is presented, and the importance of an ecosystem-based food web monitoring approach to the value of specimen banking is discussed.

Organochlorine contaminants in blubber of four seal species: Integrating biomonitoring and specimen banking

Blubber samples from four Alaska seal species (bearded seal, Erignathus barbatus; harbor seal, Phoca vitulina; northern fur seal, Callorhinus ursinus; ringed seal, P. hispida) were collected for inclusion in the US National Biomonitoring Specimen Bank, as well as for immediate analysis as part of the contaminant monitoring component of the US National Marine Fisheries Service's Marine Mammal Health and Stranding Response Program. The blubber samples were analyzed for organochlorine (OC) contaminants (e.g., PCB congeners, pesticides, DDTs). Results for bearded and ringed seals from the Alaska Arctic revealed low blubber concentrations of OC contaminants. Harbor seals from Prince William Sound, Gulf of Alaska, had somewhat higher blubber concentrations of OC contaminants. In contrast, northern fur seals sampled from the Pribilof Islands had blubber concentrations of certain OC contaminants that were about an order of magnitude higher than those found in the other seal species. Differences in contaminant concentrations among the Alaska seals may be explained by differences in feeding habits and migratory patterns; age or gender did not appear to account for the differences observed. The highest concentrations of OCs were found in harbor seals stranded along the northwestern US mainland, which is consistent with higher concentrations of anthropogenic contaminants being found in urban coastal areas than in more remote Arctic environments. The integration of real-time contaminant monitoring with specimen banking provides important baseline data that can be used to plan and manage banking activities. This includes identifying appropriate specimens that are useful in assessing temporal trends and increasing the utility of the banked samples in assessing chemical contaminant accumulation and relationships to biological effects.

Bioaccumulation of vanadium and other trace metals in livers of Alaskan cetaceans and pinnipeds.

Concentrations for 38 elements are routinely measured in the marine mammal liver tissues archived in the National Biomonitoring Specimen Bank (NBSB). Results show that hepatic concentrations of vanadium, selenium, silver, cadmium, and mercury are positively correlated with age for beluga whales (Delphinapterus leucas) and of vanadium, selenium, cadmium, and mercury with length for ringed seals (Phoca hispada). Many researchers have reported linear correlations of hepatic selenium, cadmium, and mercury with marine mammal age; however, there is only one other report of a linear correlation of hepatic vanadium with marine mammal age. Vanadium levels are at or below detection limits (< or = 0.01 micrograms/g) in liver tissues of U.S. east coast marine mammals from the NBSB but are present at levels ranging from 0.02 to 1.2 micrograms/g of wet weight in the tissues of Alaskan marine mammals. Although only three bearded seal (Eriganthus barbatus) and three bow-head whale (Balaena mysticetus) liver samples have been analyzed, hepatic vanadium levels also increased with animal size for these species. The presence of relatively high levels of vanadium in the livers of these Alaskan animals may reflect a unique dietary source of vanadium, a unique geochemical source of vanadium, or anthropogenic input to the Alaskan marine environment.

Trace element concentrations in cetacean liver tissues archived in the National Marine Mammal Tissue Bank

The National Biomonitoring Specimen Bank (NBSB), a collaborative project of several U.S. government agencies, includes marine mammal tissues collected for the Alaska Marine Mammal Tissue Archival Project (AMMTAP) and the National Marine Mammal Tissue Bank (NMMTB). Tissues were collected from 139 animals representing 13 species of marine mammals from around the U.S. Recently, concentrations for up to 30 elements in liver tissues of nine long-finned pilot whales (Globicephala melas), six harbor porpoises (Phocoena phocoena), and four white-sided dolphins (Lagenorhynchus acutus) from the NMMTB were measured using instrumental neutron activation analysis. Results from analyses of these tissues are presented, compared with results for liver tissues from other marine mammals from the AMMTAP, and compared with published values.

Relationship of silver with selenium and mercury in the liver of two species of toothed whales (odontocetes)

Liver specimens archived in the National Biomonitoring Specimen Bank from beluga whales, Delphinapterus leucas, and from Alaska and pilot whales, Globicephala melas, from the North Atlantic were analysed for silver, selenium and total mercury. Silver concentrations in beluga whales were one to three orders of magnitude higher than the concentrations in pilot whales and those reported elsewhere for other marine mammals. The concentrations of silver in the livers of beluga whales were the same or in some instances higher than the concentrations of selenium or mercury. Like mercury, silver was positively correlated with selenium in both pilot and beluga whales. This suggests a possible role for selenium in the accumulation and storage of silver in both species of whales, and raises questions about the potential for silver at such high concentrations to affect radical-scavenging enzyme systems in these marine mammals.

Considerations in the design of an environmental specimen bank: experiences of the National Biomonitoring Specimen Bank Program.

Since 1979 the National Institute of Standards and Technology (NIST) has been involved in environmental specimen banking activities as part of the National Biomonitoring Specimen Bank (NBSB). These activities have focused on the development of procedures for the collection, processing, analysis, and long-term storage of a variety of environmental specimens including: human liver, mussels and oysters, fish tissue (liver and muscle), marine mammal tissues (blubber, liver, and kidney), and marine sediments. The experiences of the NBSB can provide valuable information to assist in the design of new specimen bank efforts. Based on the experiences of the NBSB, the issues that should be addressed in the design and operation of a valid specimen bank program are presented.

Considerations in the Design of an Environmental Specimen Bank: Experiences of the National Biomonitoring Specimen Bank Program

Considerations in the Design of an Environmental Specimen Bank: Experiences of the National Biomonitoring Specimen Bank Program

Current activities within the National Biomonitoring Specimen Bank

The National Institute of Standards and Technology (NIST) has been involved in biological environmental specimen banking activities since 1979. These activities, which are known collectively as the National Biomonitoring Specimen Bank (NBSB), include the banking of a variety of specimens (human liver, sediment, mussels/oysters, fish tissue and marine mammal tissues) from several different projects supported by different government agencies. The two most recent projects, the Alaska Marine Mammal Tissue Archival Project (AMMTAP) and the National Marine Mammal Tissue Bank (NMMTB), focus on the collection, banking and analysis of marine mammal tissues and they are part of a comprehensive plan to address marine mammal monitoring, specimen banking and quality assurance of analytical measurements associated with contaminant analyses in marine mammals.

Effective tools for the trace element characterization of tissues: neutron activation analysis and voltammetry

The National Biomonitoring Specimen Bank at the National Institute of Standards and Technology applies a variety of techniques for extensive characterization of banked samples. To determine a large number of trace elements in small samples at low levels, instrumental neutron activation analysis has been combined with voltammetry. The two methods produce high quality data for thirty pollutant and biological trace elements. Results on archived specimens of human livers and intercomparisons of the two methods are reported.

Alaskan marine mammal tissue archival project

A project to establish an archive of Alaskan marine mammal tissues was conceived in 1987 to be a part of the National Biomonitoring Specimen Bank (NBSB). Protocols and field collection of marine mammals, long-term storage, and analysis are summarized in this paper. Instrumental neutron activation analysis has been used for an initial evaluation of trace element content in samples of northern fur seal (Callorhinus ursinus) from the Pribilof Islands. The findings agree with previously observed trace element levels in northern fur seals. The archived specimens can be used in future studies when comparison of past and present pollution levels are needed.