Biobanking in the COVID-19 Era and Beyond: Part 1. How Early Experiences Can Translate into Actionable Wisdom.

The era of COVID-19 has brought about a number of novel challenges for the global biobanking community. To better position the biobanking community to cope with current and future challenges, the International Society for Biological and Environmental Repositories (ISBER) COVID-19 Response Task Force was convened to identify needs and gaps in biobanking tools (existing resources that support good practice), for example, standards, best practices, business, etc. and to make recommendations to benefit the community. Toward these goals, the Task Force assembled a set of questions to explore individual biobanks' experiences, with emphasis on identification of key challenges and approaches, including tools employed. A survey was designed with the use of these questions and administered by ISBER. This article presents a summary of the aggregated data obtained from the survey responses, illustrating some of the major issues encountered and identifying which tools the survey respondents found most useful. In particular, this article focuses on the challenges identified during the early months of the COVID-19 era. Recommendations are provided to support biobank emergency preparedness for the future, address lessons learned, and propose solutions to bridge identified gaps. The analysis and the complete survey dataset will also inform the larger Task Force goal to develop specific tool recommendations.

Spiking and homogenization of biological matrices for production of reference materials using cryogenic processes.

Biological reference materials (RMs) are essential for quality assurance, traceability of measurement results and for method validation. When addressing new measurement questions or emerging regulatory issues, rigorous large-scale CRM production may not be time efficient or economically practical using current production methods. By amending a relatively small matrix batch with a compound(s) of interest at the homogenization step, the National Institute of Standards and Technology (NIST) can create a custom material on an "as-needed" basis and circumvent the time delay inherent in large-batch production, thereby generating a fit-for-purpose, rapid-response RM. Here, Coho salmon (Oncorhynchus kisutch) was cryohomogenized and spiked with an aquaculture antibiotic and antibiotic metabolite. The resultant material was analyzed using liquid chromatography-high resolution tandem mass spectrometry (LC-HRMS/MS) to determine the effectiveness of the amendment technique in a fresh-frozen matrix by assessing homogeneity and accuracy to the target concentration (e.g. mass fraction). Target mass fractions were achieved for both spike components, with RSDs below 5% in replicate measurements of each compound (n = 8). The stability of the spiked compounds was assessed one year post-production and mass fractions were stable, within 1-6% of the initial measurement results, indicating minimal change to the amended analyte concentrations over time. The results support this method as a promising new technique for custom, small-batch RM generation.

Feasibility of using the National Marine Mammal Tissue Bank for retrospective exploratory studies of perfluorinated alkyl acids.

Perfluorinated alkyl acids (PFAAs) have been used for 50+ years in materials such as stain-resistant treatments for paper and clothing, lubricants, and foam fire extinguishers. PFAAs are characterized by a fully fluorinated alkyl chain with a terminal acid group. Their long half-lives and ubiquitous environmental distribution create considerable concern for wildlife and human exposure. There is interest in examining temporal trends of PFAAs using the National Marine Mammal Tissue Bank (NMMTB), but NMMTB tissues are frozen and cryohomogenized in polytetrafluoroethylene (PTFE)-based materials. Because PTFE supplies may leach PFAAs into samples, this study mimicked collection, processing and storage steps of NMMTB samples and measured PFAA leaching to determine the feasibility of using this sample archive for PFAA temporal trends. We also explored concentrations in Atlantic white-sided dolphin (Lagenorhynchus acutus, WSDs) and rough-toothed dolphin (Steno bredanensis, RTDs) blubber (n=3 and 0) and liver (n=48 and 12, respectively). The materials used in NMMTB protocols may add up to 0.968ng/g perfluorooctanoic acid (PFOA), 0.090ng/g perfluorononanoic acid (PNFA), and 0.221ng/g perfluorooctane sulfonate (PFOS) to each archived sample. Leaching of PFNA and PFOS from supplies compared to dolphin levels was negligible, but PFOA contributions were substantially higher than levels found in most dolphin liver samples. Therefore, monitoring PFOA temporal trends from the NMMTB would require careful consideration. RTDs had significantly higher levels of PFOS and PFNA than WSDs. Both species have similar life history, trophic status, and foraging behaviors in deep pelagic waters, so differences could be from latitudinal variation in contamination. RTDs stranded in Florida; WSDs stranded farther north mostly in Massachusetts. Juveniles had significantly higher levels of PFOS and PFNA than adults in both species, suggesting growth dilution as they approach maturity. PFOS significantly decreased after 2001 in both species as expected based on changes in production.

Organohalogen Contaminants and Vitamins in Northern Fur Seals (Callorhinus ursinus) Collected During Subsistence Hunts in Alaska.

During native subsistence hunts from 1987 to 2007, blubber and liver samples from 50 subadult male northern fur seals (Callorhinus ursinus) were collected on St. Paul Island, Alaska. Samples were analyzed for legacy persistent organic pollutants (POPs), recently phased-out/current-use POPs, and vitamins. The legacy POPs measured from blubber samples included polychlorinated biphenyl congeners, DDT (and its metabolites), chlorobenzenes, chlordanes, and mirex. Recently phased-out/current-use POPs included in the blubber analysis were the flame retardants, polybrominated diphenyl ethers, and hexabromocyclododecanes. The chemical surfactants, perfluorinated alkyl acids, and vitamins A and E were assessed in the liver samples. Overall, concentrations of legacy POPs are similar to levels seen in seal samples from other areas of the North Pacific Ocean and the Bering Sea. Statistically significant correlations were seen between compounds with similar functions (pesticides, flame retardants, vitamins). With sample collection spanning two decades, the temporal trends in the concentrations of POPs and vitamins were assessed. For these animals, the concentrations of the legacy POPs tend to decrease or stay the same with sampling year; however, the concentrations of the current-use POPs increased with sampling year. Vitamin concentrations tended to stay the same across the sampling years. With the population of northern fur seals from St. Paul Island on the decline, a detailed assessment of exposure to contaminants and the correlations with vitamins fills a critical gap for identifying potential population risk factors that might be associated with health effects.

Long-term stability and temporal trends of organic contaminants in four collections of mussel tissue frozen standard reference materials.

The stability of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and chlorinated pesticides in frozen mussel tissue Standard Reference Materials (SRMs) stored at -80 °C was assessed by analyzing samples of SRM 1974, SRM 1974a, and SRM 1974b Organics in Mussel Tissue (Mytilus edulis) periodically over 25 y, 20 y, and 12 y, respectively. The most recent analyses were performed during the certification of the fourth release of this material, SRM 1974c. Results indicate the concentrations of these persistent organic pollutants have not changed during storage at -80 °C. In addition, brominated diphenyl ethers (BDEs) were quantified in each of the materials during this study. The stability information is important for on-going monitoring studies collecting large quantities of samples for future analyses (i.e., formally established specimen banking programs). Since all four mussel tissue SRMs were prepared from mussels collected at the same site in Dorchester Bay, MA, USA, the results provide a temporal trend study for these contaminants over a 17 year period (1987 to 2004).

The International Environmental Specimen Banks--let's get visible.

Environmental specimen banks (ESBs) are facilities that archive samples from the environment for future research and monitoring purposes. In addition, the long-term preservation of representative specimens is an important complement to environmental research and monitoring. Today, environmental specimen banking is experiencing a renaissance due to an increase in regulatory interest in ESB biota standards and trend data. The International Environmental Specimen Bank Group (IESB) promotes the worldwide development of techniques and strategies of environmental specimen banking and the international cooperation and collaboration among national ESBs. In order to provide a current and comprehensive overview on international environmental specimen banking activities, a questionnaire was sent to the national ESBs and asked for detailed information on the respective ESBs. The results show the rich diversity of national sampling programs, including more detailed information on archived samples, sampling strategies, and studies that have already been performed in the respective countries. All ESBs completing the survey expressed a strong interest in cooperating with other ESBs on a collaborative project. The collected information of national ESBs is intended to be made publicly available.

Environmental specimen banks as a resource for mercury and mercury isotope research in marine ecosystems.

Environmental specimen banks (ESBs) have been a fundamental tool for many nations to monitor contaminant temporal and spatial trends, study fate and transport, and assess the severity and risks of pollution. The specimens archived in ESBs are among the longest time-series, most geographically robust, and highest integrity samples available for performing environmental research. Mercury (Hg) remains one of the world's most ubiquitous environmental contaminants, and ESBs have played a prominent role in Hg research. Historically this has involved measuring concentrations of Hg species in various environmental matrices, but the emerging field of Hg stable isotope research provides a new analytical approach that can augment these traditional techniques. Signatures of Hg isotope fractionation have been effectively used for source apportionment and for elucidating Hg biogeochemical cycling. As the research surrounding Hg stable isotopes continues to mature, ESBs can play a useful role in analytical quality control, provide a robust and economical sample archive to expand and diversify the inventory of Hg isotope measurements, and be used to develop and test hypotheses to evaluate whether broadly prevailing paradigms are supported. Samples archived in ESBs are available for request by external collaborators in order to perform high impact research, and should be utilized more effectively to address emerging global environmental concerns.

Spatial and temporal trends of persistent organic pollutants and mercury in beluga whales (Delphinapterus leucas) from Alaska.

Remote locations, such as the Arctic, are often sinks for persistent contaminants which can ultimately bioaccumulate in local wildlife. Assessing temporal contaminant trends in the Arctic is important in understanding whether restrictions on legacy persistent organic pollutants (POPs) have led to concentration declines. Beluga whale (Delphinapterus leucas) tissue samples were collected from two subpopulations (Cook Inlet, Alaska and the eastern Chukchi Sea) between 1989 and 2006. Several POPs (polychlorinated biphenyls (PCBs), dichlorodiphenyldichloroethane and related compounds (DDTs), chlordanes, hexachlorocyclohexanes (HCHs), chlorobenzenes, mirex, polybrominated diphenyl ethers (PBDEs) and semi-quantitatively hexabromocyclododecanes (HBCDs)) were measured in 70 blubber samples, and total mercury (Hg) was measured in 67 liver samples from a similar set of individuals. Legacy POPs (PCBs, chlordanes, DDTs, and HCHs) were the predominant organic compound classes in both subpopulations, with median concentrations of 2360ng/g lipid for Σ80PCBs and 1890 ng/g lipid for Σ6DDTs. Backward stepwise multiple regressions showed that at least one of the four independent variables (subpopulation, sampling year, sex, and animal length) influenced the POP and Hg concentrations. ΣPCBs, ΣDDTs, Σchlordanes, Σchlorobenzenes, mirex, and Hg were significantly higher in belugas from the eastern Chukchi Sea than from the Cook Inlet (p≤0.0001). In contrast, Σ8PBDE and α-HBCD concentrations were significantly lower in belugas from the eastern Chukchi Sea than from the Cook Inlet (p<0.0001). Significant temporal increases in concentrations of Σ8PBDE and α-HBCD were observed for both subpopulations (p≤0.0003), and temporal declines were seen for ΣHCHs and Σchlorobenzenes in eastern Chukchi Sea belugas only (p≤0.0107). All other POP and Hg concentrations were stable, indicating either a lagging response of the Arctic to source reductions or the maintenance of concentrations by unregulated sources. Sex and length also significantly influenced some concentrations, and these findings are discussed.

Breakfast cereal sampling study for nutritional elements.

The National Institute of Standards and Technology (NIST) has established a Dietary Supplement Laboratory Quality Assurance Program (DSQAP) in collaboration with the National Institutes of Health Office of Dietary Supplements (NIH-ODS). The DSQAP invites laboratories twice annually to participate in interlaboratory studies where participants elect to measure concentrations of nutritional and/or toxic elements as well as active and/or marker compounds. One of these studies was designed to determine the effects of material granularity and sample processing techniques on measurement variability (precision) as well as to provide participating laboratories information on their performance relative to the NIST assigned values (bias) and to the other participants (concordance). Participants were asked to determine the mass fractions of Ca, Fe, and Zn, in mg/kg, in six breakfast cereal samples. Cereal samples consisted of three ground materials (homogenized wheat, wheat, and rice), two flake materials (wheat and rice) and a partially crushed material (a wheat/rice mixture). In general, approximately 25% of the laboratories processed and analyzed the suite of six cereal materials with adequate to exemplary measurement precision. Over half of the laboratories (60%) experienced measurement issues related to only a particular type of cereal matrix or for only a single element. A small number (15%) of laboratories experienced significant sample processing or measurement problems. Future studies planned by the DSQAP may be designed to use commercial products to aid laboratories with their sampling and analytical techniques.

Mercury stable isotopes in seabird eggs reflect a gradient from terrestrial geogenic to oceanic mercury reservoirs.

Elevated mercury concentrations ([Hg]) were found in Alaskan murre (Uria spp.) eggs from the coastal embayment of Norton Sound relative to insular colonies in the northern Bering Sea-Bering Strait region. Stable isotopes of Hg, carbon, and nitrogen were measured in the eggs to investigate the source of this enrichment. Lower δ(13)C values in Norton Sound eggs (-23.3‰ to -20.0‰) relative to eggs from more oceanic colonies (-20.9‰ to -18.7‰) indicated that a significant terrestrial carbon source was associated with the elevated [Hg] in Norton Sound, implicating the Yukon River and smaller Seward Peninsula watersheds as the likely Hg source. The increasing [Hg] gradient extending inshore was accompanied by strong decreasing gradients of δ(202)Hg and Δ(199)Hg in eggs, indicating lower degrees of mass-dependent (MDF) and mass-independent Hg fractionation (MIF) (respectively) in the Norton Sound food web. Negative or zero MDF and MIF signatures are typical of geological Hg sources, which suggests murres in Norton Sound integrated Hg from a more recent geological origin that has experienced a relatively limited extent of aquatic fractionation relative to more oceanic colonies. The association of low δ(202)Hg and Δ(199)Hg with elevated [Hg] and terrestrial δ(13)C values suggested that Hg stable isotopes in murre eggs effectively differentiated terrestrial/geogenic Hg sources from oceanic reservoirs.

Regional, temporal, and species patterns of mercury in Alaskan seabird eggs: mercury sources and cycling or food web effects?

Mercury concentration ([Hg]), δ(15)N, and δ(13)C values were measured in eggs from common murres (Uria aalge), thick-billed murres (U. lomvia), glaucous gulls (Larus hyperboreus), and glaucous-winged gulls (L. glaucescens) collected in Alaska from 1999 to 2005. [Hg] was normalized to a common trophic level using egg δ(15)N values and published Hg trophic magnification factors. Egg [Hg] was higher in murres from Gulf of Alaska, Cook Inlet, and Norton Sound regions compared to Bering Sea and Bering Strait regions, independent of trophic level. We believe the Yukon River outflow and terrestrial Hg sources on the southern Seward Peninsula are responsible for the elevated [Hg] in Norton Sound eggs. Normalizing for trophic level generally diminished or eliminated differences in [Hg] among taxa, but temporal variability was unrelated to trophic level. Normalizing murre egg [Hg] by trophic level improves the confidence in regional comparisons of Hg sources and biogeochemical cycling in Alaska.

Geographic differences in organic contaminants and stable isotopes (δ¹³C, δ¹5N) in thick-billed murre (Uria lomvia) eggs from Alaska.

The contents from thick-billed murre (Uria lomvia) eggs collected at four Alaskan colonies in 2002 were analyzed for organic contaminants and carbon δ¹³C) and nitrogen (δ¹5N) stable isotopes. Contaminant concentrations in the eggs varied from below detection limits to 230 ng g⁻¹ wet mass for 4,4'-DDE in one egg from St Lazaria Island in the Gulf of Alaska. Eggs from this colony generally contained higher levels of contaminants and exhibited significantly different patterns compared to eggs from the Bering and Chukchi seas. Stable isotope values also varied geographically; however, these differences appeared to be related to differences in C and N baselines in the food webs instead of differences in prey. Contaminant and stable isotope correlations were inconclusive, suggesting that better information on regional food web differences and differential offloading of contaminants and stable isotopes to the eggs must be obtained before these kinds of data can be fully incorporated into seabird egg contaminant monitoring programs.

Marine mammal blubber reference and control materials for use in the determination of halogenated organic compounds and fatty acids.

The National Institute of Standards and Technology (NIST) has a diverse collection of control materials derived from marine mammal blubber, fat, and serum. Standard Reference Material (SRM) 1945 Organics in Whale Blubber was recertified for polychlorinated biphenyl (PCB) congeners, organochlorine pesticides, and polybrominated diphenyl ether (PBDE) congeners. SRM 1945 has also been assigned mass fraction values for compounds not frequently determined in marine samples including toxaphene congeners, coplanar PCBs, and methoxylated PBDE congeners which are natural products. NIST also has assigned mass fraction values, as a result of interlaboratory comparison exercises, for PCB congeners, organochlorine pesticides, PBDE congeners, and fatty acids in six homogenate materials produced from marine mammal blubber or serum. The materials are available from NIST upon request; however, the supply is very limited for some of the materials. The materials include those obtained from pilot whale blubber (Homogenates III and IV), Blainville's beaked whale blubber (Homogenate VII), polar bear fat (Homogenate VI), and California sea lion serum (Marine Mammal Control Material-1 Serum) and blubber (Homogenate V).

Monitoring organic contaminants in eggs of glaucous and glaucous-winged gulls (Larus hyperboreus and Larus glaucescens) from Alaska.

Gull eggs have been used to monitor contaminants in many parts of the world. The Seabird Tissue Archival and Monitoring Project (STAMP) is a long-term program designed to track trends in pollutants in northern marine environments using seabird eggs. Glaucous and glaucous-winged gull (Larus hyperboreus and Larus glaucescens) eggs collected in 2005 from seven Alaskan colonies were analyzed for organic contaminants. Concentrations ranged from below detection limits to 322 ng g(-1) wet mass in one egg for 4,4'-DDE and differed among the samples collected in the Gulf of Alaska and Bering and Chukchi Seas. Chick growth and survival rates may be affected by the contaminant levels found in the eggs, but the eggs should be safe for human consumption if they are eaten in small quantities. STAMP plans to continue collecting and banking gull eggs for future real-time and retrospective analyses.

Hexabromocyclododecane in white-sided dolphins: temporal trend and stereoisomer distribution in tissues.

Hexabromocyclododecane (HBCD) is a brominated flame retardant used primarily in expanded polystyrene foams and other styrene resins. Samples of blubber (n = 57) and liver (n = 16) from Atlantic white-sided dolphins, Lagenorhynchus acutus, that stranded on the eastern coast of United States between 1993 and 2004 were obtained from the National Marine Mammal Tissue Bank (NMMTB). Blubber samples from most of these animals (n = 47) were previously analyzed for polybrominated diphenyl ethers (PBDE), polychlorinated biphenyls (PCB), and several toxaphene congeners. The three most abundant diastereomers in the technical HBCD mixture (alpha-HBCD, beta-HBCD, and gamma-HBCD) and their enantiomers were determined using liquid chromatography-triple quadrupole mass spectrometry (LC-MS/MS). alpha-HBCD was found in all blubber and liver samples while beta-HBCD and gamma-HBCD were not detected in any samples. The alpha-HBCD concentration in blubber and liver ranged from 14 ng/g wet mass (19 ng/g lipid) to 280 ng/g wet mass (380 ng/g lipid) and 0.051 ng/g wet mass (2.9 ng/g lipid) to 3.6 ng/g wet mass (140 ng/g lipid), respectively. Concentrations of alpha-HBCD were 2 to 3 orders of magnitude lower than previously reported PBDE, PCB, and toxaphene concentrations in these same animals. There was not a significant temporal trend for these compounds in white-sided dolphin blubber. The enantiomeric fractions (EF) measured in blubber and liver were not statistically different and ranged from 0.34 to 0.53. Blubber EFs were significantly correlated with both alpha-HBCD concentrations and white-sided dolphin body length. In general, concentrations of HBCDs were lower in these white-sided dolphins than in cetaceans from Western Europe.

Development and application of an ultratrace method for speciation of organotin compounds in cryogenically archived and homogenized biological materials.

An accurate, ultra-sensitive and robust method for speciation of mono, di, and tributyltin (MBT, DBT, and TBT) by speciated isotope-dilution gas chromatography-inductively coupled plasma-mass spectrometry (SID-GC-ICPMS) has been developed for quantification of butyltin concentrations in cryogenic biological materials maintained in an uninterrupted cryo-chain from storage conditions through homogenization and bottling. The method significantly reduces the detection limits, to the low pg g(-1) level (as Sn), and was validated by using the European reference material (ERM) CE477, mussel tissue, produced by the Institute for Reference Materials and Measurements. It was applied to three different cryogenic biological materials-a fresh-frozen mussel tissue (SRM 1974b) together with complex materials, a protein-rich material (whale liver control material, QC03LH03), and a lipid-rich material (whale blubber, SRM 1945) containing up to 72% lipids. The commutability between frozen and freeze-dried materials with regard to spike equilibration/interaction, extraction efficiency, and the absence of detectable transformations was carefully investigated by applying complementary methods and by varying extraction conditions and spiking strategies. The inter-method results enabled assignment of reference concentrations of butyltins in cryogenic SRMs and control materials for the first time. The reference concentrations of MBT, DBT, and TBT in SRM 1974b were 0.92 +/- 0.06, 2.7 +/- 0.4, and 6.58 +/- 0.19 ng g(-1) as Sn (wet-mass), respectively; in SRM 1945 they were 0.38 +/- 0.06, 1.19 +/- 0.26, and 3.55 +/- 0.44 ng g(-1), respectively, as Sn (wet-mass). In QC03LH03, DBT and TBT concentrations were 30.0 +/- 2.7 and 2.26 +/- 0.38 ng g(-1) as Sn (wet-mass). The concentration range of butyltins in these materials is one to three orders of magnitude lower than in ERM CE477. This study demonstrated that cryogenically processed and stored biological materials are a promising alternative to conventional freeze-dried materials for organotin speciation analysis, because these are, at present, the best conditions for minimizing degradation of thermolabile species and for long-term archival. Finally, the potential of the analytical method was illustrated by analysis of polar bear (Ursus maritimus) and beluga whale (Delphinapterus leuca) liver samples that had been collected in the Arctic and archived at the Marine Environmental Specimen Bank. Significant concentrations of butyltin compounds were found in the samples and provide the first evidence of the presence of this class of contaminant in the Arctic marine ecosystem. Figure Eye catch image.

Certification of methylmercury content in two fresh-frozen reference materials: SRM 1947 Lake Michigan fish tissue and SRM 1974b organics in mussel tissue (Mytilus edulis).

This paper describes the development of two independent analytical methods for the extraction and quantification of methylmercury from marine biota. The procedures involve microwave extraction, followed by derivatization and either headspace solid-phase microextraction (SPME) with a polydimethylsiloxane (PDMS)-coated silica fiber or back-extraction into iso-octane. The identification and quantification of the extracted compounds is carried out by capillary gas chromatography/mass spectrometric (GC/MS) and inductively coupled plasma mass spectrometric (GC/ICP-MS) detection. Both methods were validated for the determination of methylmercury (MeHg) concentrations in a variety of biological standard reference materials (SRMs) including fresh-frozen tissue homogenates of SRM 1946 Lake Superior fish tissue and SRM 1974a organics in mussel tissue (Mytilus edulis) and then applied to the certification effort of SRM 1947 Lake Michigan fish tissue and SRM 1974b organics in mussel tissue (Mytilus edulis). While past certifications of methylmercury in tissue SRMs have been based on two independent methods from the National Institute of Standards and Technology (NIST) and participating laboratories, the methods described within provide improved protocols and will allow future certification efforts to be based on at least two independent analytical methods within NIST.