Affiliation:
1. Universidade Federal de Minas Gerais
2. Fundacao Oswaldo Cruz Instituto Rene Rachou
3. Aggeu Magalhaes Institute: Instituto Aggeu Magalhaes
4. Centro de Bioengenharia de Espécies Invasoras de Hidrelétricas
5. UFMG: Universidade Federal de Minas Gerais
6. UEMG: Universidade do Estado de Minas Gerais
Abstract
Abstract
Among invasive species known to occur in South America, the bivalve mollusc Limnoperna fortunei, which the presence is linked to several environmental and economic problems. Early detection and mitigation actions are needed to limit its impact in the remaining L. fortunei-free areas. PCR-based molecular methods have become the gold standard methodology for L. fortunei detection. However, PCR-based methods require complex logistics from field sampling to laboratory processing. Thus, the use of methods that can be directly applied in the field can speed up the detection process. This work aimed to establish, for the first time, the loop-mediated isothermal amplification (LAMP) method for the detection of L. fortunei, with perspectives for in situ application. A set of primers designed for LAMP was tested for amplification of DNA from L. fortunei adult tissues and environmental samples containing bivalve larvae. The test showed a limit of detection as low as 0.01 ng of DNA obtained from adult tissue samples and a minimum reaction time of 60 min. The set of primers used seems to be specific for L. fortunei, since there was no cross-amplification with other bivalve or invasive molluscs that co-occur with the golden mussel in the same environment. The LAMP technique also proved to be efficient in amplifying DNA derived from L. fortunei larvae, demonstrating it to be a robust method regarding potential environmental reaction inhibitors. Although the results obtained here were acquired under controlled laboratory conditions, the LAMP method is a promising tool to integrate L. fortunei invasion monitoring protocols.
Publisher
Research Square Platform LLC
Reference43 articles.
1. The economic costs of biological invasions in Brazil: a first assessment;Adelino JRP;NeoBiota,2021
2. I See Golden Mussel! They are Everywhere! Environmental DNA Supports Widespread Dissemination of Limnoperna fortunei in Hydrographic Basins in the Paraná State, Brazil;Andrade PDB;Brazilian Archives of Biology and Technology,2021
3. Prediction of future risk of invasion by Limnoperna fortunei (Dunker, 1857)(Mollusca, Bivalvia, Mytilidae) in Brazil with cellular automata;Barbosa NP;Ecol Ind,2018
4. “Limnoperna fortunei (Dunker, 1857)(Mollusca, Bivalvia, Mytilidae): first record in the São Francisco river basin;Barbosa NP;Brazil ” Check List,2016
5. Testing a molecular protocol to monitor the presence of golden mussel larvae (Limnoperna fortunei) in plankton samples;Boeger WA;J Plankton Res,2007