Sensitive and accurate detection methods are critical for monitoring and managing the spread of aquatic invasive species, such as invasive Silver Carp (SC; Hypophthalmichthys molitrix) and Bighead Carp (BH; Hypophthalmichthys nobilis) near the Great Lakes. A new detection tool called environmental DNA (eDNA) sampling, the collection and screening of water samples for the presence of the target species’ DNA, promises improved detection sensitivity compared to conventional surveillance methods. However, the application of eDNA sampling for invasive species management has been challenging due to the potential of false positives, from detecting species’ eDNA in the absence of live organisms. In this dissertation, I study the sources of error and uncertainty in eDNA sampling and develop statistical tools to show how eDNA sampling should be utilized for monitoring and managing invasive SC and BH in the United States. In chapter 2, I investigate the environmental and hydrologic variables, e.g. reverse flow, that may be contributing to positive eDNA sampling results upstream of the electric fish dispersal barrier in the Chicago Area Waterway System (CAWS), where live SC are not expected to be present. I used a beta-binomial regression model, which showed that reverse flow volume across the barrier has a statistically significant positive relationship with the probability of SC eDNA detection upstream of the barrier from 2009 to 2012 while other covariates, such as water temperature, season, chlorophyll concentration, do not. This is a potential alternative explanation for why SC eDNA has been detected upstream of the barrier but intact SC have not. In chapter 3, I develop and parameterize a statistical model to evaluate how changes made to the US Fish and Wildlife Service (USFWS)’s eDNA sampling protocols for invasive BH and SC monitoring from 2013 to 2015 have influenced their sensitivity. The model shows that changes to the protocol have caused the sensitivity to fluctuate. Overall, when assuming that eDNA is randomly distributed, the sensitivity of the current protocol is higher for BH eDNA detection and similar for SC eDNA detection compared to the original protocol used from 2009-2012. When assuming that eDNA is clumped, the sensitivity of the current protocol is slightly higher for BH eDNA detection but worse for SC eDNA detection. In chapter 4, I apply the model developed in chapter 3 to estimate the BH and SC eDNA concentration distributions in two pools of the Illinois River where BH and SC are considered to be present, one pool where they are absent, and upstream of the electric barrier in the CAWS given eDNA sampling data and knowledge of the eDNA sampling protocol used in 2014. The results show that the estimated mean eDNA concentrations in the Illinois River are highest in the invaded pools (La Grange; Marseilles) and are lower in the uninvaded pool (Brandon Road). The estimated eDNA concentrations in the CAWS are much lower compared to the concentrations in the Marseilles pool, which indicates that the few eDNA detections in the CAWS (3% of samples positive for SC and 0.4% samples positive for BH) do not signal the presence of live BH or SC. The model shows that >50% samples positive for BH or SC eDNA are needed to infer AC presence in the CAWS, i.e., that the estimated concentrations are similar to what is found in the Marseilles pool. Finally, in chapter 5, I develop a decision tree model to evaluate the value of information that monitoring provides for making decisions about BH and SC prevention strategies near the Great Lakes. The optimal prevention strategy is dependent on prior beliefs about the expected damage of AC invasion, the probability of invasion, and whether or not BH and SC have already invaded the Great Lakes (which is informed by monitoring). Given no monitoring, the optimal strategy is to stay with the status quo of operating electric barriers in the CAWS for low probabilities of invasion and low expected invasion costs. However, if the probability of invasion is greater than 30% and the cost of invasion is greater than $100 million a year, the optimal strategy changes to installing an additional barrier in the Brandon Road pool. Greater risk-aversion (i.e., aversion to monetary losses) causes less prevention (e.g., status quo instead of additional barriers) to be preferred. Given monitoring, the model shows that monitoring provides value for making this decision, only if the monitoring tool has perfect specificity (false positive rate = 0%).

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Ce mémoire porte sur la mise au point de plateforme microfluidique couplée à la microscopie Raman confocale, utilisée dans des conditions d’excitation de la diffusion Raman (diffusion Raman exaltée de surface), dans le but d’obtenir une détection de très haute sensibilité d’espèces moléculaires sous écoulement dans des canaux de dimensions micrométriques. Ce travail a pour ambition de démontrer la faisabilité d’un couplage microscopie Raman/microfluidique en vue de la caractérisation in-situ et locale, des espèces et des réactions mises en jeu dans les fluides en écoulement dans les microcanaux. Nous avons utilisé un microcanal de géométrie T, fabriqué par lithographie douce, dans lequel sont injectées, à vitesse constante, des nanoparticules métalliques d’or ou d’argent dans une des deux branches du canal et une solution de pyridine ou de péfloxacine dans l’autre branche. La laminarité et la stationnarité du processus nous ont permis de cartographier la zone de mélange et de mettre en évidence l’exaltation du signal de diffusion Raman de la pyridine et de la péfloxacine, obtenue grâce aux nanoparticules métalliques, dans cette zone d’interdiffusion. L’enregistrement successif de la bande d’absorption des nanoparticules d’argent (bande plasmon) et du signal de diffusion Raman de la péfloxacine, en écoulement dans un microcanal, nous a permis d’établir un lien entre la morphologie des nanostructures métalliques, et plus précisément l’état d’agrégation des nanoparticules d’argent, et l’exaltation du signal Raman de la péfloxacine observé. Nous avons alors modifié la géométrie du canal afin d’y introduire une solution d’électrolyte (NaCl et NaNO3) et de modifier localement la charge de surface des colloïdes d’argent en écoulement. Nous avons ainsi confirmé que la modification de l’état d’agrégation des nanoparticules d’argent, induite par l’ajout contrôlé de solutions d’électrolytes, permet d’amplifier le signal SERS de la péfloxacine et d’optimiser la détection en microfluidique. Enfin, nous avons développé une seconde approche qui consistait à mettre en place une structuration métallisée des parois d’un microcanal. Nous avons ainsi démontré que la fonctionnalisation chimique de surface via un organosilane (APTES) permettait de tapisser le canal avec des nanoparticules d’argent et d’amplifier le signal Raman des espèces en écoulement dans ce même microcanal. / This thesis focuses on the development of a microfluidic platform coupled with confocal Raman microscopy, used in excitation conditions of Raman scattering (Surface enhanced Raman scattering, SERS) in order to gain in the detection sensitivity of molecular species flowing in channels of micrometer dimensions. This work aims to demonstrate the feasibility of coupling Raman microscopy / microfluidics for the in situ and local characterization of species and reactions taking place in the fluid flowing in microchannels. We used a T-shaped microchannel, made by soft lithography, in which gold or silver nanoparticles injected at constant speed, in one of the two branches of the channel and a solution of pyridine or pefloxacin in the other one. The laminar flow and the stationarity of the process allowed us to map the mixing zone and highlight the enhancement of the Raman signal of pyridine and pefloxacin, due to the metallic nanoparticles, in the interdiffusion zone. The recording of the both absorption band of the silver nanoparticles (plasmon band) and the Raman signal of pefloxacin, flowing in microchannel, allowed us to establish a link between the shape of the metallic nanostructure, and more precisely the silver nanoparticle aggregation state, and the enhancement of the Raman signal of pefloxacin observed. We then changed the channel geometry to introduce an electrolyte solution (NaCl and NaNO3) and locally modify the surface charge of the colloids. We have put in evidence that the change of the silver nanoparticle aggregation state, induced by the controlled addition of electrolyte solutions, could amplify the SERS signal of pefloxacin and thus optimizing the detection in microfluidics. At last, we established second a approach that consists in the metallic structuring of microchannel walls. This has shown that the surface chemical functionalization through organosilanes (APTES) allowed the pasting of the channel with silver nanoparticles, thus amplifying the Raman signal of the species flowing within the same microchannel.