PhD Defense Carla Pereira García
Mersor cium
Mercury (Hg) is a toxic and widespread metal. Monomethylmercury (MMHg) is a potent neurotoxic and especially worrisome due to its bioaccumulation and biomagnification through the aquatic food webs. In aquatic systems, MMHg is form mainly in sediments by prokaryotes carrying the hgcAB genes. MMHg demethylation and HgII reduction can also take place in sediments, performed by prokaryotes containing the mer genes. Operon mer has two enzymes, merA (mercuric reductase) which converts HgII to Hg0, and merB (organomercurial lyase) which is responsible for the degradation of MMHg to HgII. Methylation has been extensively studied since the discovery of the hgcAB gene cluster in 2013. However, studying MMHg demethylation in the environment has been very difficult, thus the main players involved in this process and the main factors controlling their activity remain largely unknown.
In the first chapter, I incubated sediments from the Venice Lagoon to simulate a MOSE (barrier system) closing event in two seasons, summer and autumn, and to determine the impact of tide-regulating barriers in the formation of MMHg. In summer, I found an increase in the concentration of DOC and an enrichment of proteinaceous substances and reactive humic acids, a higher MMHg concentration in the water column and higher methylation rates in incubated sediment cores after 48h. Both higher methylation rates and increased MMHg concentrations could potentially enhance bioaccumulation in the lagoon’s biota. These results are especially worrisome in the context of global change in which the lagoon will closed more frequently for longer times due to the increasing number of flooding events. Moreover, the expected raise in temperature might increase MMHg formation rates and/or prolong the warmer season during which MMHg is formed in sediments.
In Chapter 2, by combining metagenomic and metatranscriptomic data from sediments, my results shed light on the ecology and functional traits of the merA and/or merB containing prokaryotes. The MERCLUB-MAG Collection was reconstructed (with 803 MAGs) and the merAB containing MAGs were extracted (45 MAGs) to study their abundance, activity and metabolic capacities. The main findings are i) merB was more frequent and active than merA, ii) merAB containing MAGs had a high level of taxonomic novelty, although Gammaproteobacteria was the most predominant taxa and iii) the merAB containing MAGs had very diverse metabolisms.
Parallelly, the MERCLUB Culture Collection was obtained with the intention of finding the best candidate to be used in bioremediation strategies. Initially, 46 isolates were chosen for general physiological assessment, and after a selection process, the growth rates and the Hg removal capacities of three final candidates – Aeromonas sp. MERCC_1069, Pseudomonas putida MERCC_1942 and Bacillus sp. MERCC_2852 were determined. Aeromonas sp. was highly efficient in batch cultures, although Pseudomonas putida MERCC_1942 was finally selected to be optimized using the continuous culture technique to achieve maximum Hg removal in the bioremediation process. Strain MERCC_1942 grown at high growth rates presented the highest removal efficiency and thus was the proposed candidate for bioremediation.
Lastly, I used Alteromonas mediterranea ISS312, which is globally and vertically distributed throughout oceans, to study the effects of MMHg in its growth and in its gene’s expression. I sequence the genome and characterized the mer genes, showing that it has 4 different operons. Secondly, I performed a transcriptomic experiment in which I compared control (not amended) and samples amended with 5 mM of MMHg. In the amended cultures, I observed a longer lag phase in the growth curve, and differential expression of some but not all of the mer genes. Additionally, the mer genes presented very diverse co-expression patterns among them. These results indicate that this bacterial strain can be used as a model microorganism for MMHg demethylation and HgII reduction.