DNA barcoding

Austrian-Danish research team discover as many as 22 new moth species from across Europe

The last time so many previously unknown moths have been discovered at once in the best-studied continent was in 1887

*One of the newly discovered moths, Megacraspedus faunierensis, in its natural habitat in the Alps.*

Following a long-year study of the family of twirler moths, an Austrian-Danish research team discovered a startling total of 44 new species, including as many as 22 species inhabiting various regions throughout Europe.

Given that the Old Continent is the most thoroughly researched one, their findings, published in the open access journal ZooKeys, pose fundamental questions about our knowledge of biodiversity. Such wealth of new to science European moths has not been published within a single research article since 1887.

“The scale of newly discovered moths in one of the Earth’s most studied regions is both sensational and completely unexpected,” say authors Dr Peter Huemer, Tyrolean State Museum, and Ole Karsholt of the University of Copenhagen‘s Zoological Museum. To them, the new species come as proof that, “despite dramatic declines in many insect populations, our fundamental investigations into species diversity are still far from complete”.

The challenge of taxonomy

*Type locality of the new moth species Megacraspedus faunierensis, Cottian Alps, Italy.*

For the authors, it all began when they spotted what seemed like an unclassifiable species of twirler moth in the South Tyrolean Alps. In order to confirm it as a new species, the team conducted a 5-year study into the type specimens of all related species spread across the museum collections of Paris, London, Budapest and many in between.

To confirm the status of all new species, the scientists did not only look for characteristic colouration, markings and anatomical features, but also used the latest DNA methods to create unique genetic fingerprints for most of the species in the form of DNA barcodes.

What’s in a name?

A particular challenge for the researchers was to choose as many as 44 names for the new species. Eventually, they named one of the species after the daughter of one of the authors, others – after colleagues and many others – after the regions associated with the particular species. Megacraspedus teriolensis, for example, is translated to “Tyrolean twirler moth”.

Amongst the others, there is one which the scientists named Megacraspedus feminensisbecause they could only find the female, while another – Megacraspedus pacificus, discovered in Afghanistan – was dubbed “an ambassador of peace”.

Mysterious large twirler moths

All new moths belong to the genus of the large twirler moths (Megacraspedus) placed in the family of twirler moths (Gelechiidae), where the common name refers to their protruding modified mouthparts (labial palps).

The genus of the large twirler moths presents an especially interesting group because of their relatively short wings, where their wingspan ranges between 8 and 26 millimetres and the females are often flightless. While it remains unknown why exactly their wings are so reduced, the scientists assume that it is most likely an adaptation to the turbulent winds at their high-elevation habitats, since the species prefer mountain areas at up to 3,000 metres above sea level.

Out of the 85 documented species, however, both sexes are known in only 35 cases.

The scientists suspect that many of the flightless females are hard to spot on the ground. Similarly, caterpillars of only three species have been observed to date.

While one of the few things we currently know about the large twirler moths is that all species live on different grasses, Huemer and Karsholt believe that it is of urgent importance to conduct further research into the biology of these insects, in order to identify their conservation status and take adequate measures towards their preservation.

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Original source:

Huemer P, Karsholt O (2018) Revision of the genus Megacraspedus Zeller, 1839, a challenging taxonomic tightrope of species delimitation (Lepidoptera, Gelechiidae). ZooKeys 800: 1-278. https://doi.org/10.3897/zookeys.800.26292

A primer in access and benefit-sharing for DNA barcoders

New open access book provides essential background for molecular biodiversity researchers on international policy regarding use and transfer of genetic materials

Molecular biology approaches, such as DNA barcoding, have become part of the standard toolkit for a growing number of biodiversity researchers and practitioners, with an increasing scope of applications in important areas, such as environmental assessment, food inspection, disease control and public education.

Globalization and the advent of bioinformatics are rapidly changing the landscape of international scientific collaborations, which now often span multiple jurisdictions and increase the volume of international data exchange and transactions of biological materials. At the same time, researchers engaging in such partnerships are often unaware of the complex policy frameworks governing such transactions, which may carry reputational and even legal liabilities.

The United Nations Convention on Biological Diversity (1992) and its supplementary agreement, the Nagoya Protocol (ratified in 2014), are the most prominent international treaties designed to provide a legal framework for ensuring the fair and equitable sharing of the benefits arising from research activities involving genetic resources. Although often challenging and, at times, frustrating, it is important for researchers to understand the ramifications of these international agreements, to ensure that their scientific reputations are not tainted with allegations of unfair or unethical practices.

The recent book by Canadian ABS consultant and advisor to Botanic Gardens Conservation International, Kate Davis, and University of Guelph, Canada, researcher and international development expert, Alex Borisenko, offers a perspective on the ramifications of the Convention and the Nagoya Protocol on molecular biodiversity research.

Titled ‘Introduction to Access and Benefit-Sharing and the Nagoya Protocol: What DNA Barcoding Researchers Need to Know‘, it is openly available from Pensoft as an advanced book or PDF document under Creative Commons License.

This contribution is specifically geared towards researchers and practitioners working in the field of DNA barcoding – an actively developing field of biology that advances molecular tools for fast, reliable identification and discovery of species by analyzing short standardized DNA fragments, known as ‘DNA barcode regions’.

This approach, lying at the interface between genomics and biodiversity science, is creating the global knowledge base needed to assess ecosystem services and detect emerging environmental threats, while addressing the imperative of preserving the world’s biodiversity. Carrying out this mission demands close partnerships between biodiversity researchers worldwide, and also relies on large molecular facilities to provide timely, cost-effective and high-quality analytical services, thereby involving active international transactions of biological materials.

Furthermore, the utility of DNA barcoding depends on active open data sharing in ways similar to those established by the medical community for human genomic information.

The book is prefaced by the Executive Secretary of the Convention on Biological Diversity, Dr. Cristiana Pa?ca Palmer. It provides a brief introduction to the Convention and the Nagoya Protocol, and reviews some of their key legal definitions (e.g., ‘genetic resources’, ‘access’, and ‘utilization’). These definitions are considered within the context of terms more familiar to researchers (e.g., tissue samples, DNA extracts, PCR products, trace files) and their daily activities (e.g., field collecting, molecular analysis, DNA sequence assembly).

The main chapters provide further insights into the structure and function of the access and benefit-sharing mechanism at the international policy level and its possible ramifications in form of national laws and institutional requirements.

The text concludes with a set of practical guidelines for researchers and practitioners on the steps that should be taken to ensure due diligence when working with internationally-sourced biological samples. Adhering to these best practices would help build trust and sustain research collegiality among partners involved in international collaboration.

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Original source:

Davis K, Borisenko A (2017) Introduction to Access and Benefit-Sharing and the Nagoya Protocol: What DNA Barcoding Researchers Need to Know. Advanced Books. https://doi.org/10.3897/ab.e22579

A genus of European paper wasps revised for the first time using integrative taxonomy

The European and Mediterranean species of the paper wasp genus Polistes were recently revised by scientists at the SNSB-Zoologische Staatssammlung München (ZSM).

For the first time for this group scientists applied an integrative taxonomic approach which combines traditional morphological methods with modern DNA barcoding.

As a result, the researchers were able to identify a new species from Morocco. For this well-researched wasp group, this is an actual sensation.

The study is published in the open access journal ZooKeys.

The Munich researchers analysed more than 260 wasp specimens collected from across the study area with the help of DNA barcoding.

They managed to identify all species and determine their distribution. In addition, based on the genetic data, they were able to evaluate morphological characters for each species and created a completely new key for identification.

The wasps of the genus Polistes belong to the family Vespidae. The genus is represented by 17 species in Europe and the Mediterranean, with four species occurring in Germany. Within the genus, 13 species are social, with the queen overwintering and founding a new nest with up to 200 workers. Four species are parasitic and have no workers.

Although Polistes has been well-known in Central Europe for more than 200 years, knowledge of Mediterranean species has so far been scarce. Many species of the genus exhibit only subtle morphological differences and show high levels of colour variation, further complicating their identification.

An important result of this research is the separation of species of the Polistes gallicus species complex into three distinct species. Moreover, the genetic data led to the discovery of a new species, represented by a single specimen from the High Atlas Mountains in Morocco. This was an unexpected result for the researchers. The species was named Polistes maroccanus.

Another very surprising result was the discovery of high levels of genetic variation within Polistes dominula, a species commonly found in Central Europe, indicating the presence of up to three different and hitherto unrecognized species – a case requiring further investigation.

Integrative taxonomy is an approach that combines different scientific methods to reliably differentiate species. In particular, DNA barcoding has proven to be a useful technique for the identification of species and for the discovery of new species. The method allows to identify most species quickly and accurately, even those species that are difficult to identify using traditional methods based on morphological characters.

DNA barcoding uses a short gene fragment that differs in almost all species worldwide. The sequences are stored in an online database and can be used for identification. The method derives its name for being reminiscent of the barcodes similar to those found on products in supermarkets that allow quick and error-free identification at the checkout.

DNA barcoding is part of a global research initiative led by the Canadian scientist Paul Hebert from the University of Guelph. The ZSM is a project partner and involved in assembling DNA barcodes of the German animal species. In addition to ZSM researchers, scientists from Switzerland and the Netherlands contributed to the Polistes project.

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Original source:

Schmid-Egger C, van Achterberg K, Neumeyer R, Morinière J, Schmidt S (2017) Revision of the West Palaearctic Polistes Latreille, with the descriptions of two species – an integrative approach using morphology and DNA barcodes (Hymenoptera, Vespidae). ZooKeys 713: 53-112. https://doi.org/10.3897/zookeys.713.11335

Efficiency of insect biodiversity monitoring via Malaise trap samples and DNA barcoding

The massive decline of over 75% insect biomass reported from Germany between 1989 and 2013 by expert citizen scientists proves the urgent need for new methods and standards for fast and wide-scale biodiversity assessments. If we cannot understand species composition, as well as their diversity patterns and reasons behind them, we will fail not only to predict changes, but also to take timely and adequate measures before species go extinct.

An international team of scientists belonging to the largest and connected DNA barcoding initiatives (iBOL, GBOL, BFB), evaluated the use of DNA barcode analysis applied to large samples collected with Malaise traps as a method to rapidly assess the arthropod fauna at two sites in Germany between May and September.

One Malaise trap (tent-like structure designed to catch flying insects by attracting them to its walls and then funneling them into a collecting bottle) was set in Germany’s largest terrestrial protected natural reserve Nationalpark Bayerischer Wald in Bavaria. Located in southeast Germany, from a habitat perspective, the park is basically a natural forest. The second trap was set up in western Germany adjacent to the Middle River Rhine Valley, located some 485 kilometers away from the first location. Here, the vegetation is eradicated annually due to St. Martin’s fires, which occur every November. Their findings are published in the open access Biodiversity Data Journal.

DNA barcoding enables the identification of a collected specimen by comparing its BIN (Barcode Index Number) against the BOLD database. In contrast to evaluation using traditional morphological approaches, this method takes significantly less experience, time and effort, so that science can easily save up on decades of professional work.

However, having analyzed DNA barcodes for 37,274 specimens equal to 5,301 different BINs (i.e., species hypotheses), the entomologists managed to assign unambiguous species names to 35% of the BINs, which pointed to the biggest problem with DNA barcoding for large-scale insect inventories today, namely insufficient coverage of DNA barcodes for Diptera (flies and gnats) and Hymenoptera (bees and wasps) and allied groups. As the coverage of the reference database for butterflies and beetles is good, the authors showcase how efficient the workflow for the semi-automated identification of large sample sizes to species and genus level could be.

In conclusion, the scientists note that DNA barcoding approaches applied to large-scale samplings collected with Malaise traps could help in providing crucial knowledge of the insect biodiversity and its dynamics. They also invite their fellow entomologists to take part and help filling the gaps in the reference library. The authors also welcome taxonomic experts to make use of the unidentified specimens they collected in the study, but also point out that taxonomic decisions based on BIN membership need to be made within a comparative context, “ideally including morphological data and also additional, independent genetic markers”. Otherwise, the grounds for the decision have to be clearly indicated.

The study is conducted as part of the collaborative Global Malaise Trap Program (GMTP), which involves more than 30 international partners. The aim is to provide an overview of arthropod diversity by coupling the large-scale deployment of Malaise traps with the use of specimen-based DNA barcoding to assess species diversity.

Sequence analyses were partially defrayed by funding from the government of Canada through Genome Canada and the Ontario Genomics Institute in support of the International Barcode of Life project. The German Barcode of Life project (GBOL) is generously supported by a grant from the German Federal Ministry of Education and Research (FKZ 01LI1101 and 01LI1501) and the Barcoding Fauna Bavarica project (BFB) was supported by a 10-year grant from the Bavarian Ministry of Education, Culture, Research and Art.

Original source:

Geiger M, Moriniere J, Hausmann A, Haszprunar G, Wägele W, Hebert P, Rulik B (2016) Testing the Global Malaise Trap Program – How well does the current barcode reference library identify flying insects in Germany? Biodiversity Data Journal 4: e10671. https://doi.org/10.3897/BDJ.4.e10671

Surprising exotic flies in the backyard: New gnat species from Museum Koenig’s garden

Little did scientists Kai Heller and Björn Rulik expect to discover a new species in Germany’s Alexander Koenig Museum‘s garden upon placing a malaise trap for testing purposes. Not only did an unknown and strikingly coloured gnat get caught, but it turned out to be a species, which showed to have much more in common with its relatives from New Zealand. Their study is published in the open access Biodiversity Data Journal (BDJ).

While the genus, which the new dark-winged fungus gnat species belongs to, likely originates from the Australasian region, it was so far represented by only three species in Europe. None of them, however, stands out with the contrasting colouration of the presently announced fourth one.

The new gnat, called Ctenosciara alexanderkoenigi after the German museum’s founder, is described based on a single specimen caught in the framework of the German Barcode of Life Project (GBOL). Over three days, the scientists observed the flying insects getting caught in a malaise trap, placed among the predominantly non-native plants in the Alexander Koenig Museum’s garden. This tent-like structure is designed to catch flying insects. Once they fly into its walls, they get funnelled into a collecting bottle.

Upon noticing the beautiful striking colour of the fly, the two specialists were convinced they had just discovered a new to science species. Most of these flies are bright brownish, and the only other orange European dark-winged fungus gnat – almost uniformly orange. In contrast, the new species stands out with a mixture of reddish, black and yellowish-white hues. Based on the DNA-barcode match with New Zealand specimens, the authors concluded that the species must have arrived from the Australasian region in Europe quite recently.

“It is a rare occurrence, that a species from the opposite end of the world is represented by a single specimen only and it is not yet clear, whether Ctenosciara alexanderkoenigi has a permanent population in Germany or if it was only introduced casually with plants or soil,” they explain. “Probably, the species was recently introduced from the Australasian Region. If it was a permanent member of the European fauna, a striking species like this would likely have been found earlier.”

In conclusion, the scientists note that modern technologies such as the high quality photo documentation, established as a standard by the BOLD project, DNA barcodes assigned with BINs, as well as facilitated by speedy publishing, have largely aided taxonomists to build on the biodiversity knowledge.

“We believe that the rapid description of Ctenosciara alexanderkoenigi, coupled with the BDJ reviewing system, might be a robust and ground-breaking way to accelerate and stabilise taxonomy in the future,” they finish their paper.

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Original source:

Heller K & Rulik B (2016) Ctenosciara alexanderkoenigi sp. n. (Diptera: Sciaridae), an exotic invader in Germany? Biodiversity Data Journal 4: e6460. doi: 10.3897/BDJ.4.e6460