The Laurentian Great Lakes (LGL) constitute one of the largest freshwater systems in the world while providing social and economic value to two powerful nations. The spatial scale of these inland seas falls between two endpoints: small lakes and oceans. Lacustrine in many characteristics, the LGL often require a scientific approach with attributes similar to those of oceanography. There is a strong sense that within the LGL support for scientific research has not kept pace with the need for process-oriented research and that we lack basic information needed to forecast change, mitigate impacts and restore and preserve the LGL. Consequently, 58 researchers met in September 2014 and identified five "Grand Challenges for Research in the LGL": (1) How has this vast inland freshwater system responded to shifting climate in the past, and how will it respond in the future? (2) What is the current status of the most important ecosystem processes, including their variability in space and time? (3) What processes are characteristic only of large lakes, and how do the distinct habitats integrate into a whole? (4) What are the ecosystem responses to major anthropogenic forces such as nutrients and invasive species, and are these reversible? and (5) What are the small to large-scale linkages and feedbacks among societal decisions, biological systems, and physicochemical dynamics? An urgent need exists for a unified scientific voice that articulates the Grand Challenges for research in the LGL and the need for associated funding. This treatise describing the Grand Challenges develops that voice.
Loop-mediated isothermal amplification (LAMP) of aquatic invasive species environmental DNA (AIS eDNA) was used for rapid, sensitive, and specific detection of Dreissena sp. relevant to the Great Lakes (USA) basin. The method was validated for two uses including i) direct amplification of eDNA using a hand filtration system and ii) confirmation of the results after DNA extraction using a conventional thermal cycler run at isothermal temperatures. Direct amplification eliminated the need for DNA extraction and purification and allowed detection of target invasive species in grab or concentrated surface water samples, containing both free DNA as well as larger cells and particulates, such as veligers, eggs, or seeds. The direct amplification method validation was conducted using Dreissena polymorpha and Dreissena bugensis and uses up to 1 L grab water samples for high target abundance (e.g., greater than 10 veligers (larval mussels) per L for Dreissena sp.) or 20 L samples concentrated through 35 μm nylon screens for low target abundance, at less than 10 veligers per liter water. Surface water concentrate samples were collected over a period of three years, mostly from inland lakes in Michigan with the help of a network of volunteers. Field samples collected from 318 surface water locations included i) filtered concentrate for direct amplification validation and ii) 1 L grab water sample for eDNA extraction and confirmation. Though the extraction-based protocol was more sensitive (resulting in more positive detections than direct amplification), direct amplification could be used for rapid screening, allowing for quicker action times. For samples collected between May and August, results of eDNA direct amplification were consistent with known presence/absence of selected invasive species. A cross-platform smartphone application was also developed to disseminate the analyzed results to volunteers. Field tests of the direct amplification protocol using a portable device (Gene-Z) showed the method could be used in the field to obtain results within one hr (from sample to result). Overall, the direct amplification has the potential to simplify the eDNA-based monitoring of multiple aquatic invasive species. Additional studies are warranted to establish quantitative correlation between eDNA copy number, veliger, biomass or organismal abundance in the field.
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