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Acta Limnologica Brasiliensia
Acta Limnologica Brasiliensia
Thematic Section: Methods

Optimized and validated protocol to the detection of the invasive bivalve Limnoperna fortunei from eDNA plankton samples

Protocolo otimizado e validado para detecção do bivalve invasor Limnoperna fortunei de amostras de DNA ambiental de plâncton

Josiane Ribolli; Sophia Cassol; Samara Hermes Silva; Evoy Zaniboni Filho; Flávia Lucena Zacchi; Jacó Joaquim Mattos; Grasiela Fagundes Minatto Cardoso; Alex Pires de Oliveira Nuñer

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Abstract: : We optimized a methodology for plankton environmental DNA detection of the invasive golden mussel and validated it in samples from a Southern Brazil reservoir. Limnoperna fortunei is a successful invasive alien species that causes significant impacts on freshwater ecosystems. We adjusted and validated the methodology to detect L. fortunei in plankton samples, with a SYBR Green assay. Based on the standard curve analysis, the observed theoretical minimal qPCR detection level was 0.0005625 ng.µL-1 (R2 = 0.99) at a PCR quantification cycle of 14.09–29.56. We also presented a practical guide to be used in monitoring and detection of L. fortunei. The optimized protocol was efficient in detecting L. fortunei and can be used to monitor already infested environments or invasions in new environments.


biological invasion, biomonitoring, golden mussel, qPCR, reservoir


Resumo: : Nós otimizamos uma metodologia para detecção do mexilhão-dourado em amostras de DNA ambiental de plâncton e a validamos em amostras de um reservatório no sul do Brasil. Limnoperna fortunei é uma espécie exótica invasora bem-sucedida que promove impactos significativos nos ecossistemas de água doce. Nós ajustamos e validamos a metodologia para detectar L. fortunei em amostras de plâncton em reservatório, através de ensaios com SYBR Green. O nível teórico mínimo de detecção de qPCR observado, com base na análise de curva padrão, foi de 0,0005625 ng.µL-1 (R2 = 0,99) em um ciclo de quantificação de 14,09–29,56. Nós também apresentamos um guia prático a ser utilizado no monitoramento da invasão de L. fortunei em bacias hidrográficas. A metodologia ajustada foi eficiente na detecção de L. fortunei e, portanto, pode ser usada para monitorar ambientes já infestados ou invasões em novos ambientes em larga escala.


invasão biológica, biomonitoramento, mexilhão-dourado, qPCR, reservatórios


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