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Medusozoa in Portugal: impact on the ecosystems and development of DNA-based tools for the early forecasting of mass occurrences

Tomás Rodrigues1, 2, Ana Matos1, 2, Dany Domínguez-Pérez1, Joana Falcão1, 2, 3, Sónia C. Marques1, 3, Sérgio Leandro3, Daniela Almeida1 and Agostinho Antunes1, 2*
  • 1 Interdisciplinary Center for Marine and Environmental Research, Abel Salazar Institute of Biomedical Sciences, University of Porto, Portugal
  • 2 Faculty of Sciences, University of Porto, Portugal
  • 3 Centro de Ciências do Mar e do Ambiente (IPLeiria), Portugal

Medusozoa is a subphylum of Cnidaria composed of aquatic animals generally toxic with complex life cycles that include the two main body forms of cnidarians: a benthic polyp and a pelagic medusa. Medusozoans are divided into four classes: Hydrozoa (Owen, 1843) - hydras, hydroids, hydromedusae, and siphonophores; Scyphozoa (Goette, 1887) - true jellyfishes; Cubozoa (Werner, 1973) - box jellyfishes or sea wasps; and Staurozoa (Collins & Marques, 2004) - stalked jellyfishes. Some of the species encompassing this group have the capability to fast increase in abundance leading to huge impacts in fisheries, public health, tourism, industry and aquaculture. Those impacts are produced by the free-living stage of cnidarians known as “jellyfish”, influenced by some natural factors, but also induced by global warming, overfishing, eutrophication and widening of invasive species habitats. These events have been reported worldwide and the global panorama corresponds to an increase on the occurrence of this phenomena. Despite the common occurrence of these events, early warning tools are still lacking. Specifically, in Portugal, although jellyfish blooms are commonly reported by the Portuguese National Authorities, the information related to Medusozoa, their mass occurrence events and associated impacts is scarce. Thus, in order to increase the knowledge into this topic, the main aims of this work included: (i) a bibliographic review compiling for the first time a list of the Medusozoa reported in Portugal (mainland, Azores and Madeira); (ii) a collection of the genetic information available in public databases for the Class Scyphozoa representing the “true jellyfishes”, organisms able to produce mass occurrence events; and (iii) increase the genetic data in Scyphozoa. The resulting information will improve the accuracy for the characterization at the genetic level of these species and thus, be pivotal for the development of early-detection DNA-based tools for mass occurrence events produced by jellyfishes. Overall, the compilation of all information available revealed that all the four groups of Medusozoa are represented in Portuguese waters, with a total of 268 different species, specifically: 250 hydrozoans; 15 scyphozoans; 1 cubozoan; and 2 staurozoans species. The analysis of the genetic information available at the nucleotide database of National Center for Biotechnology Information (NCBI), for the 15 scyphozoan species that are reported in Portugal, showed that only Chrysaora quinquecirrha and Aurelia aurita had the complete mitochondrial genome available. Remarkably, Catostylus tagi, which is one of the most common scyphozoan species in Portugal, has only three sequences available, corresponding to three genetic markers: 18S ribosomal RNA (18S), 28S ribosomal RNA (28S), and Internal Transcribed Spacer 1 (ITS1). Considering the low information at molecular level available from C. tagi and its capability to occur in mass, herein we used different experimental protocols for the amplification of nuclear and mitochondrial genetic markers. The specimens were sampled in Tejo River on October 4th, 2018. The collected organisms were dissected and divided into triplicates of gonads and tentacles. Genomic DNA (gDNA) of those samples was extracted using the PureLink™ Genomic DNA Kit (Invitrogen, Carlsbad, CA, USA), following the protocol for Mammalian tissues. DNA was eluted in 50 μL of Elution Buffer. Genomic DNA was quantified in the spectrophotometer DeNovix® DS-11 FX and stored at -20ºC for further analyses. We selected for amplification some of the most phylogenetic informative genes (COI: cytochrome c oxidase subunit I, COIII: cytochrome c oxidase subunit III, 18S, 28S, ITS1 and ND6: NADH-ubiquinone oxidoreductase chain 6). For that, we performed Polymerase Chain Reactions (PCR) using different sets of primers pairs, previously used for Cnidarian species, namely (gene - primer pairs): COI gene - HCO1490/LCO2198 [1]; COIII gene - COIIIF/COIII3R [2]; 18S gene - 18S:primer A_F/18S:primer B_R [3] and 18SFb/18SRb [4]; 28S gene - 28S:primer F/28S:primer R [5] and 28S:LSUD1F_F/28S:D3Ca_R [6]; ITS1 - ITS1:jfITS1-5f_F/ITS1:jfITS1-3r_R [7]; ND6 gene - Nd6F/Nd6R (Frazão et al., in review). The PCR cycling conditions applied were the same described by the previously mentioned works for primer pairs sequence with the following exceptions: 28S - a pre-heating for 2 min at 95°C, followed by 30 cycles at 94°C (30 s), 56°C (30 min) and 72°C (1 min), with a final extension at 72°C for 10 min, using the set of primers 28S:LSUD1F_F/28S:D3Ca_R, we followed the program by Osborn & Rouse, 2010 [8]; ITS1 - a pre-heating for 8 min at 94°C, followed by 33 cycles at 94°C (45 s), 53.5°C (45 s) and 72°C (1 min), with a final extension at 72°C for 5 min. The results obtained originated the first submission to the NCBI of COI, COIII and ND6 sequences for C. tagi, as well as, new sequences of 18S, 28S and ITS1 genes. As future perspectives we aim to extend the sequencing of the aforementioned genetic markers to all the jellyfish species reported in Portugal. These sequences will be comprised in a database as a reference to detect/predict mass occurrences events based on molecular tools like metagenomic approaches.

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Acknowledgements

This work was supported in part by the Strategic Funding UID/Multi/04423/2019 through national funds provided by FCT and the European Regional Development Fund (ERDF) in the framework of the program PT2020, by the European Structural and Investment Funds (ESIF) through the Competitiveness and Internationalization Operational Program - COMPETE 2020 and by National Funds through the FCT under the project PTDC/MAR-BIO/0440/2014 (POCI-01-0145-FEDER-016772).

References

1. Folmer, O., Black, M., Hoeh, W., Lutz, R. & Vrijenhoek, R. (1994). DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol. Mar. Biol. Biotechnol. 3, 294–299. 2. Geller, J. B. & Walton, E. D. (2001). Breaking up and getting together: evolution of symbiosis and cloning by fission in sea anemones (genus Anthopleura). Evolution 55, 1781–1794. 3. Medlin, L., Elwood, H. J., Stickel, S. & Sogin, M. L. (1988). The characterization of enzymatically amplified eukaryotic 16S-like rRNA-coding regions. Gene 71, 491–499. 4. Leclère, L., Schuchert, P., Cruaud, C., Couloux, A. & Manuel, M. (2009). Molecular Phylogenetics of Thecata (Hydrozoa, Cnidaria) Reveals Long-Term Maintenance of Life History Traits despite High Frequency of Recent Character Changes. Syst. Biol. 58, 509–526. 5. Chen, C. A., Wallace, C. C., Yu, J. K. & Wei, N. V. (2000). Strategies for amplification by polymerase chain reaction of the complete sequence of the gene encoding nuclear large subunit ribosomal RNA in corals. Mar. Biotechnol. 2, 558–570. 6. Lenaers, G., Maroteaux, L., Michot, B. & Herzog, M. (1989). Dinoflagellates in evolution. A molecular phylogenetic analysis of large subunit ribosomal RNA. J. Mol. Evol. 29, 40–51. 7. Dawson, M. N. & Jacobs, D. K. (2001). Molecular evidence for cryptic species of Aurelia aurita (Cnidaria, Scyphozoa). Biol. Bull. 200, 92. 8. Osborn, K. J. & Rouse, G. W. (2010). Phylogenetics of Acrocirridae and Flabelligeridae (Cirratuliformia, Annelida). Zool. Scr. 40, 204–219.

Keywords: Cnidaria, Medusozoa, Scyphozoa, jellyfish, Catostylus tagi, Early forecasting, Mass occurrences, Blooms

Conference: XX Iberian Symposium on Marine Biology Studies (SIEBM XX) , Braga, Portugal, 9 Sep - 12 Sep, 2019.

Presentation Type: Poster Presentation

Topic: Ecology, Biodiversity and Vulnerable Ecosystems

Citation: Rodrigues T, Matos A, Domínguez-Pérez D, Falcão J, Marques SC, Leandro S, Almeida D and Antunes A (2019). Medusozoa in Portugal: impact on the ecosystems and development of DNA-based tools for the early forecasting of mass occurrences. Front. Mar. Sci. Conference Abstract: XX Iberian Symposium on Marine Biology Studies (SIEBM XX) . doi: 10.3389/conf.fmars.2019.08.00141

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Received: 04 Jun 2019; Published Online: 27 Sep 2019.

* Correspondence: Mx. Agostinho Antunes, Interdisciplinary Center for Marine and Environmental Research, Abel Salazar Institute of Biomedical Sciences, University of Porto, Matosinhos, Porto, 4450-208, Portugal, aantunes@ciimar.up.pt