Abstract
Zebra mussel (Dreissena polymorpha) is considered as one of the 100 most harmful IAS in the world. Traditional detection methods have limitations, and PCR based environmental DNA detection has provided interesting results for early warning. However, in the last years, the development of isothermal amplification methods has received increasing attention. Among them, loop-mediated isothermal amplification (LAMP) has several advantages, including its higher tolerance to the presence of inhibitors and the possibility of naked-eye detection, which enables and simplifies its potential use in decentralized settings. In the current study, a real-time LAMP (qLAMP) method for the detection of Dreissena polymorpha was developed and tested with samples from the Guadalquivir River basin, together with two real-time PCR (qPCR) methods using different detection chemistries, targeting a specific region of the mitochondrial gene cytochrome C oxidase subunit I. All three developed approaches were evaluated regarding specificity, sensitivity and time required for detection. Regarding sensitivity, both qPCR approaches were more sensitive than qLAMP by one order of magnitude, however the qLAMP method proved to be as specific and much faster being performed in just 9 min versus 23 and 29 min for the qPCR methods based on hydrolysis probe and intercalating dye respectively.
Highlights
Zebra mussel (Dreissena polymorpha) is considered as one of the 100 most harmful invasive alien species (IAS) in the world
D. polymorpha is considered by the International Union for Conservation of Nature (IUCN) as one of the 100 most harmful IAS in the world[13]
The addition of dimethyl sulfoxide (DMSO) promoted a significant increase in threshold time (Tt), which was concentrationdependent, being 27.9 ± 0.1 min for 5% DMSO and 42.3 ± 0.1 min for 7.5% DMSO
Summary
Zebra mussel (Dreissena polymorpha) is considered as one of the 100 most harmful IAS in the world. After one year of growth, they are already able to reproduce and, as adults, they can even survive 8–10 days out of water, under cool and humid c onditions[7,9] This fast spreading and growing of D. polymorpha prevents the few natural predators of this species from causing a steady and long-term decline in their population levels[10]. There are several strategies to control zebra mussels in industrial facilities, including mechanical, chemical and biological methods[14,15] These strategies can help eliminating the invasion temporarily, they sometimes require long shutdowns which can be very costly. These control methods cannot be directly applied to the colonized reservoirs, lakes or rivers, which remain an active point for the species. Some preventive measures currently applied to avoid the spread and dispersion of D. polymorpha include information and awareness campaigns, control and restrictions on navigation and installation of disinfection stations close to the affected water b odies[3]
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