RESUMEN
Demand for unconventional crude oils continues to drive the production of diluted bitumen (dilbit) within Western Canada, promoting increased transport volumes across the extensive 700,000 km pipeline system of Canada and the USA. Despite this vast extent of terrestrial transport, the current understanding of the behavior and fate of spilled dilbit within shallow groundwater systems is limited. To this end, oil spill experiments with a dilbit (Cold Lake Blend) and a physicochemical comparative conventional heavy crude oil (Conventional Heavy Blend) were conducted for 104 days in large soil columns (1 m height × 0.6 m diameter) engineered to model contaminant transport in the unsaturated (vadose) zone. Around two-fold greater concentrations and 6-41 % faster rates of vadose zone transport of benzene, toluene, ethylbenzene and xylenes (BTEX) and polycyclic aromatic compounds (PACs) were observed in the dilbit- compared to conventional heavy crude-contaminated columns. As determined by Orbitrap mass spectrometry, the OxSx species abundances in the acid extractable organics (AEOs) fraction of column leachate from both oil types increased over time, ostensibly due to microbial degradation of petroleum. Bioaccumulation of petroleum constituents in fathead minnow (Pimephales promelas) larvae exposed to contaminated leachate was confirmed through the induction of developmental malformations lasting up to 34 days and increased abundance of cyp1a mRNA observed throughout the experiment. Toxicity was comparable between the two oils but could not be fully attributed to metals, BTEX, PACs or AEOs, implying the presence of uncharacterized teratogens capable of being transported within the vadose zone following terrestrial dilbit and conventional heavy crude oil surface spills.
RESUMEN
It is generally believed that Bacillus thuringiensis var. israelensis (Bti) biopesticides are harmless to non-target organisms; however, new research shows controversial results. We exposed acutely and chronicallyLithobates sylvaticusandAnaxyrus americanus tadpoles until metamorphic climax to VectoBac 200G (granules) and VectoBac 1200L (aqueous suspension) at 300-20,000 ITU/L covering field-relevant concentrations and higher. The data show that the exposure parameters tested did not affect significantly the survival, total length, total weight, hepatosomatic index, gonadosomatic index, the expression of genes of interest (i.e., related to xenobiotic exposure, oxidative stress, and metamorphosis), and the intestine tissue layer detachment ofL. sylvaticusandA. americanus in a concentration-response pattern. In contrast, VectoBac 200G significantly increased the median time to metamorphosis ofL. sylvaticus tadpoles by up to 3.5 days and decreased the median by up to 1 day inA. americanus. VectoBac 1200L significantly increased the median time to metamorphosis ofL. sylvaticusandA. americanustadpoles by up to 4.5 days. Also, the exposure to VectoBac 200G and 1200L altered the intestine bacterial community composition inA. americanus at application rates recommended by the manufacturer, which led to an increase in the relative abundance of Verrucomicrobia, Firmicutes, Bacteroidetes, and Actinobacteria. Changes in the intestine microbiota might impact the fitness of individuals, including the susceptibility to parasitic infections. Our results indicate that the effect of Bti commercial products is limited; however, we recommend that Bti-spraying activities in amphibian-rich ecosystems should be kept minimal until there is more conclusive research to assess if the changes in the time to metamorphosis and microbiota can lead to negative outcomes in amphibian populations and, eventually, the functioning of ecosystems.
Asunto(s)
Bacillus thuringiensis , Microbioma Gastrointestinal , Animales , Agentes de Control Biológico , Ecosistema , Humanos , Larva , Control Biológico de VectoresRESUMEN
Mammalian acetylcholinesterase (AChE) is well-studied, being important in both cholinergic brain synapses and the peripheral nervous systems and also a key drug target for many diseases. In contrast, little is known about the structures and molecular mechanism of prokaryotic acetylcholinesterases. We report here the structural and biochemical characterization of ChoE, a putative bacterial acetylcholinesterase from Pseudomonas aeruginosa Analysis of WT and mutant strains indicated that ChoE is indispensable for P. aeruginosa growth with acetylcholine as the sole carbon and nitrogen source. The crystal structure of ChoE at 1.35 Å resolution revealed that this enzyme adopts a typical fold of the SGNH hydrolase family. Although ChoE and eukaryotic AChEs catalyze the same reaction, their overall structures bear no similarities constituting an interesting example of convergent evolution. Among Ser-38, Asp-285, and His-288 of the catalytic triad residues, only Asp-285 was not essential for ChoE activity. Combined with kinetic analyses of WT and mutant proteins, multiple crystal structures of ChoE complexed with substrates, products, or reaction intermediate revealed the structural determinants for substrate recognition, snapshots of the various catalytic steps, and the molecular basis of substrate inhibition at high substrate concentrations. Our results indicate that substrate inhibition in ChoE is due to acetate release being blocked by the binding of a substrate molecule in a nonproductive mode. Because of the distinct overall folds and significant differences of the active site between ChoE and eukaryotic AChEs, these structures will serve as a prototype for other prokaryotic acetylcholinesterases.