BIOPRESERVATION AND BIOBANKING Volume 11, Number 3, 2013 ª Mary Ann Liebert, Inc. DOI: 10.1089/bio.2013.0018

Recent Initiatives in Biodiversity Biobanking: Summary of Presentations from the ESBB 2012 Conference Jacqueline MacKenzie-Dodds,1 Ann Clarke,2 Dominik Lermen,3 Isabel Rey,4 Jonas J. Astrin,5 Ole Seberg,6 and Christian C. Oste7

Introduction

A

ccording to their respective charters, both the European, Middle Eastern, and African Society for Biopreservation & Biobanking (ESBB) and the International Society for Biological and Environmental Repositories (ISBER) focus a sizeable share of their activities on repositories involving nonhuman material. Those specific collections may unfortunately not usually attract the same level of funding, visibility, and notoriety in the biobanking realm as their human counterparts. Nevertheless, biodiversity is a vast, largely unexploited resource of direct, indirect, or potential use for human health (www.cbd.int/doc/health/cohab-policy-brief1-en.pdf ). Thus, biodiversity has recently become an important topic, as clues about understanding and elucidating health conditions affecting humans are increasingly being sought and encountered through the use of nonhuman models, be it animal, vegetal, or microbial. Also, indirect health issues and bio-economic aspects, as well as direct environmental and societal benefits in biodiversity biobanking, are often unfortunately ignored. Additionally, half of the most-prescribed drugs in the US, and an even higher fraction in developing countries, are derived from natural compounds—and many more are to be found. In biodiversity research, biobanks are especially relevant, because sample resources are frequently either very difficult to access or cannot be accessed repeatedly. The first situation arises from the remoteness of many collecting localities and the highly specialized knowledge necessary for collecting and identifying a plethora of species; the second can be due to human-induced rapid loss of populations or species. Studies about species evolution and adaptation to changing environmental conditions are some other aspects of the invaluable work done on samples from nonhuman repositories. Through skillful collection and conservation, it is becoming possible to accumulate snapshots of the status of specific species, arching back millions of years in some cases,

and to assemble a ‘‘living archive’’ about how progressive evolution has enabled living organisms to adjust to incessantly changing circumstances. In this meeting report, we will summarize some of the presentations dealing with biodiversity, which were made at the recent ESBB meeting, held in Granada (Spain) in November 2012.

The Frozen Ark Consortium ‘‘We should preserve every scrap of biodiversity as priceless while we learn to use it and come to understand what it means to humanity’’ - Prof. Edward O. Wilson, Harvard University. This presentation, delivered by Dr. Ann Clarke, University of Nottingham and Lucy Cavendish College, Cambridge, was yet another strong warning to all of us that many living species, regardless of taxa, are in danger of becoming extinct within the foreseeable future, should environmental conditions continue to deteriorate at the current pace. The IUCN (International Union for Conservation of Nature, http://www.iucn.org) Red List and the UNEP (United Nations Environment Programme, http://www.unep.org), which Dr. Clarke mentioned in her talk on several occasions, provide information that truly deserves urgent attention from all of us. It is estimated that 24% of mammals, 12% of birds, 20% of freshwater fish, and 50% each of amphibians and invertebrates are at risk of becoming extinct within the next 3 decades. Even if radical measures were implemented today to stop or at least slow down the rate of environmental conditions degradation, the ensuing benefits towards successfully protecting endangered species will not be felt in a timely fashion to prevent the majority of them becoming extinct (Fig. 1, Table 1). The Frozen Ark project, which was initiated in 1996 by the formation of a not-for-profit charity, stems from the resolve to preserve essential information by biobanking tissue, cells, and gametes from endangered species globally as a priority. Prof. Bryan Clarke, Dr. Ann Clarke, and the late Dr. Anne McLaren were the co-founding scientists of the project, which has substantially expanded since then.

1

Molecular Collections Facility, Natural History Museum, London, United Kingdom. The Frozen Ark Office, School of Biology, University Park, Nottingham, United Kingdom. 3 Fraunhofer-Institute for Biomedical Engineering, St. Ingbert, Germany. 4 Tissues and DNA Collection, MNCN-CSIC, Madrid, Spain. 5 Zoologisches Forschungsmuseum A. Koenig (ZFMK), Bonn, Germany. 6 Natural History Museum of Denmark, Copenhagen, Denmark. 7 BioScope International LLC, Barcelona, Spain. 2

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ESBB 2012 CONFERENCE

183

FIG. 1. Structure of the IUCN Red List Categories.

Table 1.

Synopsis of endangered species Critically Endangered (CR)

Group Mammals Birds Reptiles Amphibians Fishes Insects Mollusks Plants

1996/98 169 168 41 18 157 44 257 909

2000

2002

2003

2004

2006

2007

2008

2009

2010

2011

2012

180 182 56 25 156 45 222 1,014

181 182 55 30 157 46 222 1,046

184 182 57 30 162 46 250 1,276

162 179 64 413 171 47 265 1,490

162 181 73 442 253 68 265 1,541

163 189 79 441 254 69 268 1,569

188 190 86 475 289 70 268 1,575

188 192 93 484 306 89 291 1,577

188 190 106 486 376 89 373 1,619

194 189 137 498 414 91 487 1,731

196 197 144 509 415 119 549 1,821

Endangered (EN) Group Mammals Birds Reptiles Amphibians Fishes Insects Mollusks Plants

1996/98

2000

2002

2003

2004

2006

2007

2008

2009

2010

2011

2012

315 235 59 31 134 116 212 1,197

340 321 74 38 144 118 237 1,266

339 326 79 37 143 118 236 1,291

337 331 78 37 144 118 243 1,634

352 345 79 729 160 120 221 2,239

348 351 101 738 237 129 222 2,258

349 356 139 737 254 129 224 2,278

448 361 134 755 269 132 224 2,280

449 362 150 754 298 151 245 2,316

450 372 200 758 400 166 328 2,397

447 382 284 764 477 169 417 2,564

446 389 296 767 494 207 480 2,655

Vulnerable (VU) Group Mammals Birds Reptiles Amphibians Fishes Insects Mollusks Plants

1996/98

2000

2002

2003

2004

2006

2007

2008

2009

2010

2011

2012

612 704 153 75 443 377 451 3,222

610 680 161 83 452 392 479 3,331

617 684 159 90 442 393 481 3,377

609 681 158 90 444 389 474 3,864

587 688 161 628 470 392 488 4,592

583 674 167 631 681 426 488 4,591

582 672 204 630 693 425 486 4,600

505 671 203 675 717 424 486 4,602

505 669 226 657 810 471 500 4,607

493 678 288 654 1,075 478 587 4,708

497 682 351 655 1,137 481 769 4,861

497 727 367 657 1,149 503 828 4,914

Source: IUCN Red List (http://www.iucn.org)

184 The general idea is to conserve the genetic resources of the world’s endangered species. It is the animal equivalent of the ‘Millennium Seed Bank’ created by Kew Gardens to conserve the seeds of the world’s plants. In practical terms, this means collecting and saving genomic DNA, somatic cells, gametes, and embryos. Consortium members of The Frozen Ark currently hold over 48,000 samples from more than 5,500 endangered and non-endangered animal species. The goal is to complete the collection of relevant samples from at least 10,000 species by 2015, with a special emphasis on terrestrial invertebrates. All Consortium Members agree to collect samples and preserve them for the short- or long-term, to forward them to a long-term storage facility, if necessary, in ways that cause minimal degradation of DNA and loss of cell viability, to record and forward sample data details to Nottingham for the database, arrange for duplicate samples to go elsewhere for safety and, if requested, to receive samples for storage from those with samples with or without repository facilities of their own. Since The Frozen Ark Consortium was officially granted the status of Public Charity, we now all have the opportunity to pledge our support to this remarkable and essential project by making financial donations. For more information, please visit: http://www.frozenark.org.

The Cryo-Brehm Project At this point, and upon Dr. Dominik Lermen’s request, reference should be made to the presentation he made at the ESBB 2011 conference, rather than to the one he delivered at ESBB 2012. At the time, Dr. Lermen presented in detail the Cryo-Brehm Project, an initiative stemming from two Institutes from the Fraunhofer-Gesellschaft in Germany, the Institute for Biomedical Engineering IBMT in St. Ingbert and the Research Institution for Marine Biotechnology EMB in Lu¨beck. Cryo-Brehm is a member of The Frozen Ark Consortium and therefore fully adheres to the goals and SOPs set forth by the Consortium. In addition, continuous improvement of home-grown protocols on cell isolation and culture enables the project to accommodate the varying needs of different animal species. Cryo-Brehm is a generation spanning project focusing on archiving as many vertebrate species as possible, in particular endangered species, with a minimum of ten individuals of both sexes per species. Due to the involvement of two institutions, collected samples are stored at two separate locations: in Lu¨beck, for samples coming from institutions located in the Northern part of Germany, and in St. Ingbert for samples collected in the Southern part of the country. One half of the samples of each institution are transferred as a backup to the institute’s counterpart to increase security. In terms of the types of samples collected, the emphasis is placed on primary somatic cells, and on adult stem and progenitor cells, which, on the basis of genomic data from the species of origin, might be reprogrammed into induced pluripotent stem (iPS) cells. This line of advanced molecular research, combining genomics and proteomics approaches, has been made possible by the innovative ultra-low temperature storage technology developed by the Fraunhofer-IBMT. Specialized vials, fitted with 2-D matrix barcode, RFID and memory chip, were designed to store cells suspensions at - 196C (LN2 tanks). To

MACKENZIE-DODDS ET AL. avoid warming of frozen samples, sample handling is done in a specific semi-automated cryo-workbench developed for this project, which includes a controlled-rate freezer. At present, the combined collections of the Cryo-Brehm project feature samples representing more than 100 species, for a total of over 6000 aliquots. For more information about the Cryo-Brehm Project, and the innovative cryostorage technology developed by the Fraunhofer-IBMT, please visit: http://www.emb.fraunhofer.de/en/Uebersichtsindex/ cellbank_cryo-brehm.html http://www.ibmt.fraunhofer .de/en/Fields-of-work/ibmt-biophysics-cryotechnology.html http://www.ibmt.fraunhofer.de/en/Fields-of-work/ibmt-cell biology-applied-virology.html

The Natural History Museum (London) Another key member of The Frozen Ark Consortium was represented at ESBB 2012 by Ms. Jackie MacKenzie-Dodds, who delivered a comprehensive presentation about the goals and aspirations of NHM, with regard to both existing and new collections. Researching the contents of biodiversity sample archives from the past enables us to manage the environment more sustainably now and in the future. With respect to newly acquired samples, careful preparation starting at the point of collection in the field will enable the preservation of a variety of functional cellular components (known and unknown at this time), extending to whole clonal viable cell lines (e.g., pluri-/multi or even toti-potent stem cells) from endangered or extinct species (see Cryo-Brehm Project). Society may well need to exploit all these resources for conservation and habitat restoration wherever possible in the future, to maintain or restore a healthy biosphere in the face of habitat loss and mass extinctions of species across Earth, thereby managing food and water security, emerging and neglected diseases, and public health for generations to come. At present, Natural History Museums (NHMs) worldwide hold a wealth of biological resources from all known nonhuman species, from root to leaf-tip of the Tree of Life (ToL), from bacteria and fungi to plants and vertebrates, collected over centuries from across all known habitats and environments across the globe, representing an irreplaceable chronology of samples and specimens informing us of biodiversity patterns over time, including effects of climate change and global warming effects over recent decades, and where many of these resources are now unobtainable (extinct or endangered species, or from areas of civil conflict or political instability, or where permits to collect are withheld). NHM Biodiversity biobanks are broadly composed of three main collections strands: Traditional Collections, Archival Molecular Collections, and New Molecular Collections.

Traditional Collections (70 million samples at NHM London) ‘‘Wet’’ in liquid preservatives (formalin fixed in ethanol/ industrial methylated spirits), and ‘‘dry’’ skins, feathers, bones, pinned specimens, and herbarium sheets collected over the last 2 centuries (comprising the ‘‘ancient’’ to ‘‘archival’’ DNA range). These collections are not usually described as ‘‘biobanks’’ in the modern sense: they are mainly used as reference/voucher specimens (including Types, the original specimen from which the species was first described)

ESBB 2012 CONFERENCE for morphological characterization, and therefore destructive sampling is kept to an absolute minimum. But they also contain valuable ‘‘molecular’’ (including genetic) resources, although these are often significantly degraded by the original preservation processes and subsequent storage conditions. However, the importance of the information they still hold cannot be overstated, and unlocking this still largely untapped resource (e.g., extracting, repairing, and sequencing archival DNAs) is an important focus in many NHMs worldwide today.

Molecular Collections- Archival and New (approximately 2 million samples at NHM London) Archival molecular collections. Mostly from research projects over the last 3–4 decades containing a wide variety of plant and animal tissues and extracts (usually DNAs, many of which are still held in individual nonstandard ad hoc formats in freezers distributed around the institution), sub-sampled from whole voucher specimens (new freshly field collected and old archival) held in the traditional collections. This archive contains valuable molecular derivatives that are usually much less degraded than from the traditional collections, but require spending a lot of time to sort, evaluate, consolidate. and reformat to make accessible to the wider scientific community. New molecular collections. Highest ‘‘genomic’’ quality genetic resources from specimens, tissue sub-samples, and extracts from freshly field collected specimens kept as cold and/or dry as possible from point of collection and transported rapidly for deposition into modern biobank facility storage (-80C, LN2 tanks, with silica gel in humidity/O2% controlled cabinets, or other new ambient temperature storage systems in development). In new centralized molecular collection facilities (e.g., NHM London), field specimen data capture systems and storage vessel formats are standardized, high throughput compatible (e.g., mass data import via scanable 2-D barcoded cryo-tubes), and rapidly retrievable from the biobank for users. Inventoried samples are linked to vouchers held in the traditional collections and/or tissue or molecular vouchers held in the central facility. Genome quality data and samples are linked via a global virtual repository (GGBN, Global Genome Biodiversity Network). Challenges abound in the immediate future for institutions such as NHM London, both from an institutional as well as from a scientific/technical standpoint. Although museum biodiversity biobanks hold invaluable bio-resources for the future welfare of society, they remain relatively poorly funded, especially compared with most medical biobanks. This situation has been exacerbated in recent years by the global economic downturn and recessions across the EU, where funding of nonmedical biobanks has generally not been prioritized and core funding cut. The resulting worldwide shortage of trained taxonomists and curators in museums and herbaria especially, who provide accurate species identifications, is an increasing problem. Overcoming the challenges of sustainable resourcing for biodiversity biobank infrastructure and annual facility operating costs, as well as culture change from recently increased restrictions and regulations from global policy makers with respect to use of all biological material containing genetic resources (CBD’s Nagoya Protocol 2010, and ABS, equivalent to longer established ethics, ownership, IPR,

185 and consent policies in the medical world) is particularly challenging at this time for biodiversity biobanks. There is a strong sense of urgency in light of current environmental crises to preserve these resources and the information they contain before they disappear forever. Several issues are being addressed with on-going studies in biodiversity biobanks worldwide to meet the needs of 21st century biodiversity biobank clients, customers and investors: a ‘‘translational research’’ approach to turn academic research into practical benefits and their realization and application in the working biodiversity biobank. First, the optimization of field collection protocols to future-proof biodiversity molecular collections by safely transporting and preserving the highest quality possible genomic resources and/or cellular functionality/viability for the future. Second, the optimization and standardization of minimally destructive DNA/RNA extraction protocols (including highthroughput and automated robotic workstation) from new freshly field collected or frozen tissues and traditional collections. This work is being done partly in collaboration with the EU SYNTHESYS II Joint Research Activities. Third, the optimization and cost efficiency of long-term (‘‘museumscale’’, i.e., hundreds of years) storage protocols for biodiversity and museum/herbarium ‘‘molecular collections’’, such as tissues and cellular/genetic extracts (DNA/RNA/ protein) across the Tree of Live (ToL). Empirical data is being collected, from a museum/herbarium context, which will be used to inform future best practice for long-term storage of precious biodiversity resources. Lastly, the utilization of fragmented/damaged archival genetic resources: museums and herbaria hold invaluable collections, often irreplaceable DNA/RNA resources, which have been highly damaged by traditional preservation methods. Understanding DNA damage to museum archival specimens: biochemical changes, how to interpret remaining information (extract DNA and sequence) and repair DNAs where possible (NHM London’s project with GE/Whatman FTA: in preparation). Next generation sequencing technologies will play an increasingly important role here, and may well supersede attempts to restore damaged DNA samples in museum specimens. Reduce/ prevent further damage to samples and specimens; advice on future preservation protocols for morphological specimens where molecular analysis is required downstream. For additional information on these comprehensive development programs, please visit: http://www.nhm.ac.uk/ nature-online/biodiversity/index.html.

The MNCN-CSIC Initiative and the ToL Project ‘‘The Tree of Life Web Project (ToL) is a collaborative effort of biologists and nature enthusiasts from around the world. On more than 10,000 World Wide Web pages, the project provides information about biodiversity, the characteristics of different groups of organisms, and their evolutionary history (phylogeny)’’. Over the course of history, scientific collections at Natural History Museums have been used basically to work on taxonomy and phylogeny. The main objective of the Museum collections is to preserve the knowledge of biological diversity ex situ (from morphology to DNA), and to answer the following: How many different species exist? How are they recognized as different? and How have they diversified or what has their evolutionary history (phylogeny) been?

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MACKENZIE-DODDS ET AL.

To answer these questions, researchers have been using molecular techniques (DNA) since the 1980s, as Dr. Isabel Rey, Curator of the Tissues and DNA Collection at the MNCN-CSIC, explained in detail during her presentation. The Museo Nacional de Ciencias Naturales (MNCN-CSIC), based in Madrid, maintains the most comprehensive Natural History collections in Spain, both in size (7.8 million specimens of 38,589 types) and scientific and historical importance (since 1771). The collections are part of the Vice-direction of Collections and Documentation, which is divided into two main categories: Library and Documentation Unit and the Natural History Collections Unit. Under the Library and Documentation Unit there are three sub-collections: 1) Archive and Documental Restoration, 2) Library, and 3) Animal Sounds Library and Mediateca MNCN-CSIC. The Natural History Collections Unit comprises ten collections: B B B B B B B B B B B

Invertebrates (insects and mollusks excepted) Malacology (mollusks) Entomology Amphibians and Reptiles Ichthyology Birds and Mammals Tissues and DNA Geology Paleontology of Vertebrates and Prehistory Paleontology of Invertebrates and Botanic Decorative Arts and Archaeology of Scientific Instruments

Table 2 lists some details about the storage methods used in the Tissues and DNA collection. The MNCN-CSIC collection of tissue and DNA contains 300,000 specimens and has been housed at centralized facilities in Madrid since the year 2000. Samples in the collection are assigned an individual voucher number, which appear in Genebank, where more of 7000 sequences belonging to MNCNCSIC’s preserved samples can currently be accessed. In this collection, internal quality control on DNA yield from different preservation methods is performed every 5 years (only with tissues at present). Today the collection includes 52,100 specimens in its catalogue, with over 80,000 samples from 4000 species already classified and available to the scientific community. Twenty of these samples of tissue and DNA are either type specimens, part of type series, or name-bearing specimens. A database of 12,445 specimens is available from the Global Biodiversity Information Facilities (GBIF) (http://www.gbif.es). Another 250,000 (approximately) samples are also deposited and are currently in the process of being classified. The popularity of this collection is growing and the number of users requiring tissues or extracted DNA is steadily increasing. To date, 10,500 samples have been lent to national and international research projects, all of them with Table 2.

the required permits when they belonged to species included in annexes of CITES (Convention on International Trade in Endangered Species, http://www.cites.org). For more information about Dr. Rey’s work and about the MNCN-CSIC, please visit: http://www.mncn.csic.es.

The Global Genome Biodiversity Network (GGBN) Professor Ole Seberg, Natural History Museum of Denmark, delivered a presentation about the Tree of Life (ToL) concept (Fig. 2), and the subsequent initiative, the Global Genome Biodiversity Network (GGBN), an offspring of the Global Genome Initiative (GGI), which started in 2011. To place things in perspective, Professor Seberg initially referred to the original concept of the ‘‘Tree of Life’’ as proposed by Charles Darwin in ‘‘On the Origin of Species’’ (1859). ‘‘Our classifications will come to be, as far as they can be so made, genealogies; and will then truly give what may be called the plan of creation.’’- Charles Darwin Biodiversity traditionally includes three main components: B

B

B

Genetic diversity: refers to the genetic variation that occurs among members of the same species Species diversity (taxonomic diversity): refers to the variety of species Ecosystem diversity: refers to the variety of biological communities found on Earth.

Briefly stated, all species have genetic diversity, and all ecosystems have species diversity; and species themselves are united and classified by their relationships in the tree of life. The richer the diversity of life, the greater the opportunity for medical discoveries, economic development, and adaptive responses to climate change. Well-known examples such vinblastine and vincristine (from Catharanthus roseus, used in treatment of leukemia and Hodgkin’s lymphoma), and galantamine (from Galanthus nivalis, palliative for Alzheimer’s disease) were also mentioned in Professor’s Seberg presentation. Therefore, the variety of life is our insurance policy. Human activity has reduced natural habitats dramatically and continues to do so at an alarmingly high rate,

Storage Methods Used at the MNCN-CSIC Tissues and DNA Collection

Dried material

Dry (GE Whatman FTA card) Silica Freeze dried (lyophilized) Liquid preserved 70% (final) Alcohol (96%) DMSO buffer, EDTA buffer Frozen Deep freezers (-80C)

Tissues/DNA Tissues Tissues/DNA Tissues Tissues Tissues/DNA

FIG. 2. By Kelly M. Houle. Inspired from a drawing by Darwin’s in his ‘‘B’’ notebook ( July 1837) (http://www.illuminatedorigin.com).

ESBB 2012 CONFERENCE simultaneously driving a steadily increasing extinction. Based on a conservative estimate, there are 10–14 million species on Earth, only 1.75 million of which are known. To preserve just a synoptic, global collection of the entire genome of these 1.75 million species is a formidable task. From there, Professor Seberg went on to describe in detail the GGI and the GGBN projects, starting with comments on the background of those initiatives. A significant proportion of current biodiversity research relies on access to ‘‘genome-quality’’ tissue or DNA samples. It is therefore imperative to employ the best standards and practices in the care of such collections and to provide access to them using norms that promote research while respecting the owner’s rights as described in the Convention on Biological Diversity and the Nagoya protocol. A first step in implementing the Global Genome Initiative (GGI) took place at a workshop with participants from leading biorepositories, biorepository networks, research organizations, and biodiversity information management organizations in 2010. The aim of this workshop was the development of a global network of biorepositories that would encourage the use of best practices, standards, and accessibility. The workshop was attended by representatives from organizations in Africa, Australia, Europe, and North, Central, and South America. In light of the challenges that organizations face, and recognizing that a collaborative approach is necessary to preserving the world’s biodiversity, the workshop participants agreed to form the Global Genome Biodiversity Network (GGBN). A steering committee was named to guide the development of the network and three task forces were formed to address data standards, policies and practices, and marketing and outreach. GGBN’s working vision is a global network of wellmanaged collections of genome quality tissues samples from across the Tree of Life, benefiting society through biodiversity research, development and conservation. The working mission of GGBN is to foster collaborations among repositories of molecular biodiversity in order to ensure quality standards, improve best practices, secure interoperability, and harmonize exchange of material in accordance with national and international legislation and conventions. The goals of GGBN are as follows: 1) Provide genomequality samples from across the Tree of Life for research, training, and product development, thereby contributing to the conservation of global genetic diversity for generations to come. 2) Provide open access to a global data management system hosting the aggregated primary specimen data and metadata for all the member institutions. 3) Develop standards for sharing DNA and tissue information. 4) Develop best practices related to management and stewardship of genomic samples and their derivatives, including appropriate access and benefit sharing (ABS). 5) Promote targeted preservation of genomic samples representing a synoptic sample of Life on Earth. 6) Recruit partners with different regional and taxonomic focus, to preserve the global genetic diversity in a concerted effort. Thus, it is important to keep in mind that GGBN is not: B B B B B

Focused environmental sampling (e.g., NEON) Conservation focused (e.g., WWF or CI) Duplicating gamete, zygote, or seed repositories An effort to sequence all genomes A gene sequencing facility

187 B

Targeting microbes/viruses, human tissues, model species, crops, or diseases

One of the main expected outcomes of this initiative is to identify collection gaps across ToL. Members will be expected to adopt standards for sharing DNA/tissue information and policies and best practice guidelines. There will also be a special emphasis on the members’ website (data portal), since it will provide access to aggregated information on genomic samples, metadata, DNA, images, and publications. For more information about GGBN, please visit: http://www.ggbn.org.

The Barcode of Life Initiative (CBOL/iBOL/GBOL) Dr. Jonas J. Astrin, Zoologisches Forschungsmuseum A. Koenig (ZFMK), Bonn (Germany), presented a very convincing argument concerning the urgent need to coordinate more efficiently the activities of the biobanks with those of DNA barcoding projects. In the light of the importance of taxonomy in the current biodiversity crisis, the first point he emphasized focused on what is recognized as the ‘‘taxonomic impediment.’’ The taxonomic impediment results from gaps in our taxonomic knowledge, accompanied by the decrease in numbers of taxonomists and natural history collection curators. An estimated number of 5000 taxonomists face around 1,800,000 described and another 6,000,000 to 18,000,000 undocumented species. Without an increased focus on biological taxonomy, the discipline of delimiting and reidentifying meaningful biological units, there are no adequate means to monitor, manage, and use biodiversity. This implies major or minor restrictions for areas like conservation and natural resource management, forestry, agriculture, aquaculture, food control, pest management, human and veterinary medicine, forensics, pharmacy, ecology, and customs. Molecular techniques enable routine (mass) identifications of unknown samples, the demand for which exceeds by far the currently available capacities of the taxonomic workforce. Dr. Astrin went on to describe how DNA barcoding works and what benefits can be derived from this approach. DNA barcoding is the molecular reidentification of species using globally standardized markers/protocols. Indeed, the molecular characters involved in DNA barcoding offer a universal, standardized means of sampling and analysis, are applicable to every body part or fragment (or to extracellular DNA), and to every developmental stage or sex. Most importantly, barcoding can be automated and parallelized, which makes it fast, cost-efficient and opens up the possibility to work on mixed samples (precluding the laborious and specific step of sorting specimens) using next generation sequencing. There is a substantial worldwide interest in the DNA barcoding approach, to the point where the initiative has become a ‘‘Big Science’’ project: 200 organizations from 50 countries and 6 continents and thousands of scientists are collaborating on setting up the pivotal barcoding reference database, contributing hundreds of thousands samples annually. Barcoding projects qualify as one of the most valuable sources for biobank specimens of wild organisms, thanks to,

188 among other aspects, the high level of specimen diversity (going far beyond the model organism stage) and to a thorough taxonomic coverage, including deep and highquality metadata. Barcoding projects also fulfill one of the key requirements in biodiversity biobanking: they make complete specimen vouchers available in morphological collections that are cross-linked to the biobank, thus warranting subsequent validation and re-investigation of the specimens (this is one of the reasons why it makes sense to run biodiversity biobanks at established natural history collections such as museums or botanical gardens, in the same facility as the associated morphological collections). Looking at it from the other perspective of this mutualism, it becomes obvious that barcoding also benefits greatly from biobanking. Apart from the molecular vouchering of barcoding samples and the reproducibility of results, biobanking offers the expansion of barcoding datasets with samples from other projects. As with any other project, biobanking increases the visibility of molecular barcoding samples. Most importantly, however, proper biobanking offers ’scalability’ of barcode samples for future studies, making it possible to adapt to new standards (additional markers, nuclear or organellar genomes). Sample collection, identification, and subsampling are by far the most time- and cost-intensive steps in barcoding projects, and biobanks can make it unnecessary to go through this painful process all over again. Nevertheless, there currently remain some issues with the archiving of barcoding samples, and professional standards are not always applied to all aspects of molecular sample storage (this may have its basis in the way taxonomy has been traditionally carried out and in the previously rather low, but now rising awareness for the importance of molecular subsamples). Indeed, the availability of molecular samples for research is often low, although projects like the DNA Bank Network (http://www.dnabank-network.org/) or GGBN (see above) give considerable reason for hope. With regard to storage conditions, labeling, retrieval, monitoring, emergency plans and legal aspects (BMTAs, ABS), standard operating procedures are not always adopted. In conclusion, biobanks should be aware that DNA barcoding projects can be valuable sample sources. They should implement the proper procedures to guarantee the long-term and high-quality storage of barcoding samples, thereby consolidating their role as a key infrastructure for biodiversity-centered research and the many disciplines dependent on it. The chance to do so is now: once the barcoding reference database is assembled, mixed samples without reference samples (of low value for biobanking except ESBs) will prevail in barcoding. For further information on Dr. Astrin’s work, please visit: http://biobank.zfmk.de. Regarding the International Barcode of Life (iBOL) project, please visit: http://www.ibol.org.

MACKENZIE-DODDS ET AL. For an example of a national barcoding campaign: http:// www.bolgermany.de/

Conclusion In closing, it is appropriate to go back to the basic tenets of both of our societies, ISBER and ESBB. In essence, one of the main goals for both organizations is to preserve samples and gather information about all living organisms, even though ancient DNA analysis enables us to acquire valuable information about already extinct species, including humanoid ones. An intriguing parallelism could be drawn between nonhuman and human biodiversities, in particular in the context of nonhuman endangered species and human rare diseases. In both cases, the research focus lies on very small numbers of beings. In both cases, the challenge is to raise society’s awareness and commitment to become engaged in preserving and studying those beings. In the nonhuman context, this is necessary in order, among other reasons, to better understand how the rest of our ecosystem functions and how evolution works. We will never know exactly what and how much we do not know, unless proper means are made available to continue expanding our collective knowledge. In these times of widespread budget cuts, what does it mean in practical terms? Should we focus only on the ‘‘top of the clade’’ in the case of endangered species, even though the concept of studying a model organism will never encompass all the existing biodiversity and could therefore be considered irrelevant? Should we likewise revisit the threshold of penetrance of rare diseases, to decide which are ‘‘worth’’ studying? It is comforting to realize that initiatives such as The Frozen Ark, iBOL, and GGBN all aim at generating detailed information about biodiversity and that approaches originally developed with the human species in mind are now making their way into other branches of the tree of life. The developing confluence between human and nonhuman knowledge databases will hopefully foster a more harmonious co-existence between all species on our globally endangered beautiful Blue and Green Planet.

Address correspondence to: Dr. Christian C. Oste Department of Business Development BioScope International LLC Carrer Entenca 202 Barcelona 08029 Spain E-mail: [email protected]

Recent initiatives in biodiversity biobanking: summary of presentations from the ESBB 2012 Conference.

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