Bioremediation Potential of Select Bacterial Species for the Neonicotinoid Insecticides, Thiamethoxam and Imidacloprid

The neonicotinoid insecticides, including thiamethoxam (THM) and imidacloprid (IMI), have become increasingly favored in the past decade due to their specificity as insect neurotoxicants. However, neonicotinoids have been implicated as a potential contributing factor in Colony Collapse Disorder (CCD), the widespread disappearance of honey bees, which affects produce production on a global scale. The environmental persistence of neonicotinoids underscores the importance of developing a sustainable, ecologically-friendly remediation technique to remove residual insecticides from the environment. The present study characterizes the neonicotinoid bioremediation potential of six bacterial species: Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas aeruginosa, Alcaligenes faecalis, Escherichia coli, and Streptococcus lactis. In Phase I, we evaluated the utilization of IMI or THM as the sole carbon or nitrogen source by P. fluorescens, P. putida, and P. aeruginosa. All three species were better able to utilize THM over IMI as their sole carbon or nitrogen source, and better growth was noted when THM was used as the sole nitrogen source compared to the sole carbon source. Thus, further studies proceeded with THM only. In Phase II, we assessed the kinetics of THM removal from aqueous media by the six species. Significant (p<0.0001) reductions in 70 mg/L THM concentration were observed for P. fluorescens (67%), P. putida (65%), P. aeruginosa (52%), and A. faecalis (39%) over the 24-day study period, and for E. coli (60%) and S. lactis (12%) over the 14-day study period. The amount of time required to remove 50% of the THM in the media (T50) was: 12 days (d) (E. coli), 18 d (P. fluorescens), 19 d (P. putida), and 23 d (P. aeruginosa). Neither A. faecalis nor S. lactis achieved 50% removal during the study periods. The THM removal by all species followed a first-order kinetic reaction and half-lives were calculated accordingly. HPLC chromatograms of P. fluorescens, P. putida, and E. coli cultures revealed that as the area of the THM peak decreased over time, the area of an unidentified metabolite peak increased. In Phase II, we sought to characterize this metabolite and the overall metabolic efficiency of these three species. Maximal THM removal occurred at 30{degrees}C for all bacterial species assessed. Identification of the metabolite is currently underway, which will allow determination of whether the metabolite is less toxic than the parent compound, a prerequisite for this remediation technique to be viable. If the metabolite is found to be less hazardous than THM, further testing will follow to evaluate the use of this bioremediation technique in the field.