From Echinococcus to Fasciola

Prof. Lightowlers providing encouragement for effective recombinant vaccines against worms

Headquarters in Madrid

Instituto de Recursos Naturales y Agrobiología de Salamanca

Approaching the Institute, the Director’s influence is highly visible in the bright green entrance and surrounds, leaving the visitor in no doubt as to the direction of the research- biology, natural resources and agriculture.

In the Director’s Office, Mar tells me about her career to date – involving periods spent in Switzerland, Brazil and then Switzerland again before returning to Spain and the Institute at Salamanca. During the time spent in these various postings she built up a strong portfolio of scientific achievements involving (among other topics) multidisciplinary projects on Echinococcus  (a tapeworm causing hydatid disease) and Dirofilaria  (the “American” heartworm of dogs).

From Echinococcus to Fasciola
Mar’s involvement with a multidisciplinary team on Echinococcus enabled her to develop her first biobank.

It was while working on Echinococcus with a group of clinicians (human and veterinary) in the FP7 HERACLES consortium that Mar set up a biobank – allowing for systematic archiving and cataloguing not only of parasite isolates but also of patient samples, and of clinical data including imaging. Logically, then, her role in PARAGONE has involved using her expertise in Biobanking to provide such a searchable and retrievable archive of Fasciola (liver fluke isolates), together with RNA, cDNA and gDNA from each isolate.

Both the Echinococcus and Fasciola biobanks are managed by Institute Technician Maria Gonzalez. Intriguingly, Maria started out her scientific career as a geologist, but has clearly relished the leap from minerals to metazoans.

Mar and Maria spent some time showing me both the hardware and software required for their biobanks. The larger Echninococcus biobank comes with two bespoke searcheable databases, ERCE and CYSTRACK, this last administered from the Institute, but accessible online to partners and others with custom levels of access. The Fasciola bank has about 150 isolates to date, and so is manageable via Excel worksheets

The hardware is similar for both banks, and includes appropriate racked space in -80OC freezers, specific 96-well plates with lockable lids (more about those lids later!), and tubes which fit into the wells. Each plate has an alpha-numeric code and a bar code, and each tube has an alphanumeric code on the side, as well

When a Fasciola isolate arrives, the worm is divided into two portions along its longitudinal axis. One half is stored as it is (usually in RNA-later) in a coded tube, and the other is used to extract RNA and gDNA and prepare cDNA. All samples from one isolate are usually stored next to each other in the same plate. To date, there have been about 150 Fasciola isolates deposited in the PARAGONE biobank – comprising both field isolates, and lab-strain flukes from vaccine trials, from Argentina, Belgium, Ireland, Spain and Uruguay. These have been used to examine variation in target candidate vaccine genes – results to date indicate that these are highly conserved among isolates – good news for vaccinologists.

Biobanks in Veterinary Science

Biobanks in Veterinary Science

Biobanks in Veterinary Science

Biobanks can be used to store a wide variety of host/pathogen samples, readily archived and linked to clinical data, images, epidemiological information and population outcomes. It is important to have standard procedures for submission to biobanks. While there are strict standards laid down for human biobanks, this is not so in veterinary medicine.

We move to talking about the general usefulness of biobanks, and the comparative lack of regulation/standards for animal/veterinary biobanks as compared with those involving human samples. Mar advises that there are a number of masters’ level courses – including one in the University of Salamanca – available for those wishing to start up biobanks. In terms of usefulness – I imagine the prospect of having to ditch samples (parasite isolates, sera etc.) after a certain number of years, in the absence of a proper archiving system and a biobank manager, will be familiar to many. The IRNASA Fasciola biobank is a valuable resource both for PARAGONE researchers during the lifetime of the project, and also beyond it – how variable are those new antigens yet to be discovered?

Individual tubes have both and alphanumeric and a QR code

Individual tubes have both and alphanumeric and a QR code

In an era when “big data” dominates biology, and when advances in NGS and other “omics” technology almost daily expand the information we can retrieve from stored samples, the use of biobanks to systematically store isolates, samples, clinical 2 records and other data in large multinational projects such as PARAGONE seems a nobrainer. It is a very tangible asset developed with EU funding – so please heed Mar’s pleas for deposition of more isolates from as many places, host species, and circumstances as possible. Of course – it also works the other way around – Partners can request material stored in the bank – it will be but the work of a moment for Maria to locate it with her trusty scanner.

So now back to those locking lids. While opening the freezer to show me the racking system – a gremlin in the room ensured that the lid of a plate had not, in fact, been locked properly. There was as a result – how shall I put it – a spraying of 96 (thankfully well-sealed) tubes onto the floor of the freezer room – 94 were recaptured fairly easily – the last two took a bit of work but were eventually retrieved from under a freezer – so rest easy- all banked samples are safe.

Mar and Maria

Mar and Maria

The Fasciola biobanking work that is part of the H2020 PARAGONE project has already been instrumental in establishing that the vaccine targets for this parasite do not seem to vary between isolates from different geographic regions. Of course, an essential objective of PARAGONE is to ensure the sustainability of resources and knowledge developed after the project has finished. The biobank, thanks to Mar and Maria, will provide access to parasite isolates, RNA, cDNA and genomic DNA for the research community now and into the future. Mar would be delighted to receive samples, or indeed requests for material, from PARAGONE researchers, or others working on Fasciola, in order to make best use of this valuable resource. You can contact her at mmar.siles@irnasa.csic.es

As in much of the world, efforts are ongoing in Spain to remedy gender inequalities in science, and to make sure the contribution of women is both facilitated and recognised. Mar and Maria are certainly helping to make this happen.