Temperature, phytoplankton density and bacteria diversity drive the biotransformation of micropollutants in a lake ecosystem.

For most micropollutants (MPs) present in surface waters, such as pesticides and pharmaceuticals, the contribution of biotransformation to their overall removal from lake ecosystems is largely unknown. This study aims at empirically determining the biotransformation rate constants for 35 MPs at different periods of the year and depths of a meso-eutrophic lake. We then tested statistically the association of environmental parameters and microbial community composition with the biotransformation rate constants obtained. Biotransformation was observed for 14 out of 35 studied MPs for at least one sampling time. Large variations in biotransformation rate constants were observed over the seasons and between compounds. Overall, the transformation of MPs was mostly influenced by the lake's temperature, phytoplankton density and bacterial diversity. However, some individual MPs were not following the general trend or association with microorganism biomass. The antidepressant mianserin, for instance, was transformed in all experiments and depths, but did not show any relationship with measured environmental parameters, suggesting the importance of specific microorganisms in its transformation. The results presented here contribute to our understanding of the fate of MPs in surface waters and thus support improved risk assessment of contaminants in the environment.

Effect of light on the transformation of BDE-47 by living and autoclaved cultures of Microcystis flos-aquae and Chlorella vulgaris.

Polybrominated diphenyl ethers (PBDEs) are ubiquitous and toxic contaminants found in high concentrations in watercourses, and are not well removed by conventional wastewater treatment facilities. This study aimed to evaluate the removal and transformation of BDE-47, one of the environmentally predominant PBDE congener, by a green alga (Chlorella vulgaris) and a cyanobacterium (Microcystis flos-aquae) under different light conditions. Living and autoclaved cultures were exposed to BDE-47 at a concentration of 10 µg L-1 for 7 days. Both species removed >90% of BDE-47 very shortly after spiking. Light intensity affected the transformation of BDE-47 in living cultures of both species, since 5 to 11 times more debromination products were measured at a light intensity of 100 µmol photons m-2 s-1 than at 20 µmol photons m-2 s-1. Living cultures of M. flos-aquae transformed BDE-47 at a rate of 0.22 day-1 while no transformation was observed in the respective autoclaved cultures. On the contrary, both living and autoclaved cultures of C. vulgaris had similar BDE-47 transformation rates of 0.05-0.06 day-1. Debromination of BDE-47 was a predominant transformation pathway in cultures of C. vulgaris, with two times higher BDE-28 concentrations measured than in M. flos-aquae, while hydroxylation was more dominant with the cyanobacterium. Most BDE-47 and its debromination product BDE-28 were found on the cell surface of both species. These results reveal that different transformation mechanisms were involved in C. vulgaris and M. flos-aquae cultures and confirm the importance of species selection for the removal of PBDEs from contaminated environments.

Designing and Engineering Methylorubrum extorquens AM1 for Itaconic Acid Production.

Methylorubrum extorquens (formerly Methylobacterium extorquens) AM1 is a methylotrophic bacterium with a versatile lifestyle. Various carbon sources including acetate, succinate and methanol are utilized by M. extorquens AM1 with the latter being a promising inexpensive substrate for use in the biotechnology industry. Itaconic acid (ITA) is a high-value building block widely used in various industries. Given that no wildtype methylotrophic bacteria are able to utilize methanol to produce ITA, we tested the potential of M. extorquens AM1 as an engineered host for this purpose. In this study, we successfully engineered M. extorquens AM1 to express a heterologous codon-optimized gene encoding cis-aconitic acid decarboxylase. The engineered strain produced ITA using acetate, succinate and methanol as the carbon feedstock. The highest ITA titer in batch culture with methanol as the carbon source was 31.6 ± 5.5 mg/L, while the titer and productivity were 5.4 ± 0.2 mg/L and 0.056 ± 0.002 mg/L/h, respectively, in a scaled-up fed-batch bioreactor under 60% dissolved oxygen saturation. We attempted to enhance the carbon flux toward ITA production by impeding poly-ß-hydroxybutyrate accumulation, which is used as carbon and energy storage, via mutation of the regulator gene phaR. Unexpectedly, ITA production by the phaR mutant strain was not higher even though poly-ß-hydroxybutyrate concentration was lower. Genome-wide transcriptomic analysis revealed that phaR mutation in the ITA-producing strain led to complex rewiring of gene transcription, which might result in a reduced carbon flux toward ITA production. Besides poly-ß-hydroxybutyrate metabolism, we found evidence that PhaR might regulate the transcription of many other genes including those encoding other regulatory proteins, methanol dehydrogenases, formate dehydrogenases, malate:quinone oxidoreductase, and those synthesizing pyrroloquinoline quinone and thiamine co-factors. Overall, M. extorquens AM1 was successfully engineered to produce ITA using acetate, succinate and methanol as feedstock, further supporting this bacterium as a feasible host for use in the biotechnology industry. This study showed that PhaR could have a broader regulatory role than previously anticipated, and increased our knowledge of this regulator and its influence on the physiology of M. extorquens AM1.

Sensitivity of Scenedesmus obliquus and Microcystis aeruginosa to atrazine: effects of acclimation and mixed cultures, and their removal ability.

Atrazine is an herbicide frequently detected in watercourses that can affect the phytoplankton community, thus impacting the whole food chain. This study aims, firstly, to measure the sensitivity of monocultures of the green alga Scenedemus obliquus and toxic and non-toxic strains of the cyanobacteria Microcystis aeruginosa before, during and after a 30-day acclimation period to 0.1 µM of atrazine. Secondly, the sensitivity of S. obliquus and M. aeruginosa to atrazine in mixed cultures was evaluated. Finally, the ability of these strains to remove atrazine from the media was measured. We demonstrated that both strains of M. aeruginosa had higher growth rate-based EC50 values than S. obliquus when exposed to atrazine, even though their photosynthesis-based EC50 values were lower. After being exposed to 0.1 µM of atrazine for 1 month, only the photosynthesis-based EC50 of S. obliquus increased significantly. In mixed cultures, the growth rate of the non-toxic strain of M. aeruginosa was higher than S. obliquus at high concentrations of atrazine, resulting in a ratio of M. aeruginosa to total cell count of 0.6. This lower sensitivity might be related to the higher growth rate of cyanobacteria at low light intensity. Finally, a negligible fraction of atrazine was removed from the culture media by S. obliquus or M. aeruginosa over 6 days. These results bring new insights on the acclimation of some phytoplankton species to atrazine and its effect on the competition between S. obliquus and M. aeruginosa in mixed cultures.

Tolerance of cyanobacteria to the toxicity of BDE-47 and their removal ability.

Polybrominated diphenyl ethers are ubiquitous and toxic contaminants in aquatic environments. The effect of polybrominated diphenyl ether BDE-47 on five species of cyanobacteria, along with their removal ability was investigated. Four species, namely Synechocystis sp., Oscillatoria planctonica, Microcystis flos-aquae and Nostoc sp., were exposed to BDE-47 at concentrations ranging from 0.05 to 1.0 mg L-1 for 14 days, while the exposure time for Pseudanabaena sp. was 30 days. The first four species were very tolerant to BDE-47 while growth and photosynthesis of Pseudanabaena were significantly inhibited by BDE-47 at concentrations over 0.1 mg L-1. However, this species could recover from the toxicity of high concentrations of BDE-47 after 30 days of exposure, indicating the development of some "resistance" after pre-exposure to 1.0 mg L-1 BDE-47. The "resistant" cells had a higher growth rate, photosynthesis and glutathione S-transferase activity than normal Pseudanabaena cells. The sensitivity of Pseudanabaena to BDE-47 toxicity was affected by its initial filament density, with cultures having a low filament density (2.3 × 106 filaments mL-1) being up to 14-15 times more sensitive than cultures with a high filament density (13 × 106 filaments mL-1). All cyanobacteria could remove 70-82% of BDE-47 in their media, with more than 60% of BDE-47 accumulated in cells. This is the first study showing the high tolerance of different cyanobacteria species to BDE-47 toxicity and their removal ability. The study also revealed that the sensitive Pseudanabaena could acquire a "resistance" to BDE-47, which was transferred to the next generation.

Proteomic analysis reveals differences in tolerance to acid rain in two broad-leaf tree species, Liquidambar formosana and Schima superba.

Acid rain (AR) is a serious environmental issue inducing harmful impacts on plant growth and development. It has been reported that Liquidambar formosana, considered as an AR-sensitive tree species, was largely injured by AR, compared with Schima superba, an AR-tolerant tree species. To clarify the different responses of these two species to AR, a comparative proteomic analysis was conducted in this study. More than 1000 protein spots were reproducibly detected on two-dimensional electrophoresis gels. Among them, 74 protein spots from L. formosana gels and 34 protein spots from S. superba gels showed significant changes in their abundances under AR stress. In both L. formosana and S. superba, the majority proteins with more than 2 fold changes were involved in photosynthesis and energy production, followed by material metabolism, stress and defense, transcription, post-translational and modification, and signal transduction. In contrast with L. formosana, no hormone response-related protein was found in S. superba. Moreover, the changes of proteins involved in photosynthesis, starch synthesis, and translation were distinctly different between L. formosana and S. superba. Protein expression analysis of three proteins (ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit, ascorbate peroxidase and glutathione-S-transferase) by Western blot was well correlated with the results of proteomics. In conclusion, our study provides new insights into AR stress responses in woody plants and clarifies the differences in strategies to cope with AR between L. formosana and S. superba.

Alteration of plant physiology by glyphosate and its by-product aminomethylphosphonic acid: an overview.

It is generally claimed that glyphosate kills undesired plants by affecting the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) enzyme, disturbing the shikimate pathway. However, the mechanisms leading to plant death may also be related to secondary or indirect effects of glyphosate on plant physiology. Moreover, some plants can metabolize glyphosate to aminomethylphosphonic acid (AMPA) or be exposed to AMPA from different environmental matrices. AMPA is a recognized phytotoxin, and its co-occurrence with glyphosate could modify the effects of glyphosate on plant physiology. The present review provides an overall picture of alterations of plant physiology caused by environmental exposure to glyphosate and its metabolite AMPA, and summarizes their effects on several physiological processes. It particularly focuses on photosynthesis, from photochemical events to C assimilation and translocation, as well as oxidative stress. The effects of glyphosate and AMPA on several plant physiological processes have been linked, with the aim of better understanding their phytotoxicity and glyphosate herbicidal effects.

Algal fluorescence sensor integrated into a microfluidic chip for water pollutant detection.

We report the first miniaturized fluorescent sensor based on algae, with an organic light emitting diode (OLED) and an organic photodetector (OPD) integrated into a microfluidic chip. The blue emission OLED was used as the excitation source, while a blend of PTB3/PC(61)BM was used for the fabrication of the organic photodetector. Excitation and emission color filters based on acid/base dyes and a metal complex were developed and assembled with the organic optoelectronic components in order to complete the fluorescent detection system. The detection system was then integrated in a microfluidic chip made from (poly)dimethylsiloxane (PDMS). The complete sensor is designed to detect algal fluorescence in the microfluidic chamber. Algal chlorophyll fluorescence enables evaluation of the toxicity of pollutants like herbicides and metals-ions from agricultural run-offs. The entirely organic bioassay here presented allowed detection of the toxic effects of the herbicide Diuron on Chlamydomonas reinhardtii green algae that gave 50% inhibition of the algae photochemistry (EC(50)) with a concentration as low as 11 nM.

Temperature-dependent sensitivity of growth and photosynthesis of Scenedesmus obliquus, Navicula pelliculosa and two strains of Microcystis aeruginosa to the herbicide atrazine.

The temperature-dependent sensitivities of two algal species and two strains of cyanobacteria to the photosynthesis-inhibiting herbicide atrazine were evaluated in order to understand how the interaction between acclimation temperature and herbicide will affect growth and photosynthesis of aquatic microorganisms. The green alga Scenedesmus obliquus, the diatom Navicula pelliculosa and a toxic and non-toxic strain of Microcystis aeruginosa were acclimated to three different temperatures (10, 15 and 25°C) and exposed to five concentrations of the herbicide atrazine (0-0.15µM) for 72h. Growth, photosynthetic yields, energy fluxes within photosystem II and pigment content were then measured as potential responses to each treatment. With the exception of N. pelliculosa, the toxicity of atrazine was higher when microorganisms were acclimated to lower temperatures. N. pelliculosa was not only the most tolerant to atrazine, but also had a similar sensitivity to this herbicide at every temperature. The observed differences in growth sensitivity to atrazine at low temperature are associated with the ability of algae and cyanobacteria to cope with high excitation pressure, by increasing its protective carotenoid content and non-photochemical energy dissipation. Our results demonstrate that future guidelines for the protection of aquatic life should consider water temperature as an important factor influencing the toxicity of atrazine to aquatic microorganisms.