The establishment of invading organisms in natural ecosystems is one of the most serious environmental issues. In Brazil, the invasive species Limnoperna fortunei (Dunker, 1857), the golden mussel, is a mollusk capable of causing major changes in water systems, generating social and economic impacts, given its biofouling capacity. Limnoperna fortunei can easily block pipes and heat exchangers in the water systems of hydroelectric power plants due to its ability to strongly adhere to the substrate using its byssus -a bundle of filaments secreted by these animals. Therefore, the early detection of this invader is essential for management actions to be immediate, in order to control population growth rate at the beginning of the invasive process, preventing this environment from serving as a source for new infestations. The implantation of a method that integrates the active monitoring of prioritized areas, laboratory techniques, including molecular biology methods, and the sharing of hydrographic data between basin managers and users for early detection of the presence of species in Brazilian waters appears as an efficient option to prevent and control invasions.
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.
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