Anne-Marie Delort

A total of 480 microorganisms collected from 39 clouds sampled at the Puy de Dôme station (alt. 1465 m; 45°46′19′′ N, 2°57′52′′ E; Massif Central, France) were isolated and identified. This unique collection was screened for biosurfactant (surfactants of microbial origin) production by measuring the surface tension (σ) of the crude extracts, comprising the supernatants of the pure cultures, using the pendant drop technique. The results showed that 41 % of the tested strains were active producers (σ < 55 mN m⁻¹), with 7 % being extremely active (σ < 30 mN m⁻¹). The most efficient biosurfactant producers (σ < 45 mN m⁻¹) belong to a few bacterial genera (Pseudomonas and Xanthomonas) from the Υ-Proteobacteria class (78 %) and a yeast genus (Udeniomyces) from the Basidiomycota phylum (11 %). Some Bacillus strains from the Firmicutes phylum were also active but represented a small fraction of the collected population. Strains from the Actinobacteria phylum in the collection examined in the present study showed moderate biosurfactant production (45<σ < 55 mN m⁻¹). Pseudomonas (Υ-Proteobacteria), the most frequently detected genus in clouds, with some species issued from the phyllosphere, was the dominant group for the production of biosurfactants. We observed some correlations between the chemical composition of cloud water and the presence of biosurfactant-producing microorganisms, suggesting the “biogeography” of this production. Moreover, the potential impact of the production of biosurfactants by cloud microorganisms on atmospheric processes is discussed.

A total of 450 bacteria and yeast strains isolated from cloud waters sampled at the puy de Dôme station in France (1465 m) were screened for their ability to produce siderophores. To achieve this, a high-throughput method in 96-well plates was adapted from the CAS (chrome azurol S) method. Notably, 42% of the isolates were siderophore producers. This production was examined according to the phyla of the tested strains and the type of chelating functional groups (i.e., hydroxamate, catechol, and mixed type). The most active bacteria in the clouds belong to the γ-Proteobacteria class, among which the Pseudomonas genus is the most frequently encountered. γ-Proteobacteria are produced in the majority of mixed function siderophores, such as pyoverdines, which bear a photoactive group. Finally, siderophore production was shown to vary with the origin of the air masses. The organic speciation of iron remains largely unknown in warm clouds. Our results suggest that siderophores could partly chelate Fe(III) in cloud waters and thus potentially impact the chemistry of the atmospheric aqueous phase.

Within cloud water, microorganisms are metabolically active and, thus, are expected to contribute to the atmospheric chemistry. This article investigates the interactions between microorganisms and the reactive oxygenated species that are present in cloud water because these chemical compounds drive the oxidant capacity of the cloud system. Real cloud water samples with contrasting features (marine, continental, and urban) were taken from the puy de Dôme mountain (France). The samples exhibited a high microbial biodiversity and complex chemical composition. The media were incubated in the dark and subjected to UV radiation in specifically designed photo-bioreactors. The concentrations of H₂O₂, organic compounds, and the ATP/ADP ratio were monitored during the incubation period. The microorganisms remained metabolically active in the presence of^●OH radicals that were photo-produced from H₂O₂. This oxidant and major carbon compounds (formaldehyde and carboxylic acids) were biodegraded by the endogenous microflora. This work suggests that microorganisms could play a double role in atmospheric chemistry; first, they could directly metabolize organic carbon species, and second, they could reduce the available source of radicals through their oxidative metabolism. Consequently, molecules such as H₂O₂would no longer be available for photochemical or other chemical reactions, which would decrease the cloud oxidant capacity.

The biodegradation of the most abundant atmospheric organic C ₁ to C ₄ compounds (formate, acetate, lactate, succinate) by five selected representative microbial strains (three Pseudomonas strains, one Sphingomonas strain, and one yeast strain) isolated from cloud water at the puy de Dôme has been studied. Experiments were first conducted under model conditions and consisted of a pure strain incubated in the presence of a single organic compound. Kinetics showed the ability of the isolates to degrade atmospheric compounds at temperatures representative of low-altitude clouds (5°C and 17°C). Then, to provide data that can be extrapolated to real situations, microcosm experiments were developed. A solution that chemically mimicked the composition of cloud water was used as an incubation medium for microbial strains. Under these conditions, we determined that microbial activity would significantly contribute to the degradation of formate, acetate, and succinate in cloud water at 5°C and 17°C, with lifetimes of 0.4 to 69.1 days. Compared with the reactivity involving free radicals, our results suggest that biological activity drives the oxidation of carbonaceous compounds during the night (90 to 99%), while its contribution accounts for 2 to 37% of the reactivity during the day, competing with photochemistry.

Aminopolycarboxylic acids are ubiquitous in natural waters and wastewaters. They have the ability to form very stable water-soluble complexes with many metallic di- or trivalent ions. The iron complex nitrilotriacetic acid-Fe(III) (FeNTA) has been previously shown to increase drastically the rate of photo- and biodegradation of 2-aminobenzothiazole, an organic pollutant, by Rhodococcus rhodochrous . For this paper, the fate of FeNTA was investigated during these degradation processes. First, it was shown, using in situ ¹ H nuclear magnetic resonance, that the complex FeNTA was biodegraded by Rhodococcus rhodochrous cells, but the ligand (NTA) alone was not. This result indicates that FeNTA was transported and biotransformed inside the cell. The same products, including iminodiacetic acid, glycine, and formate, were obtained during the photo- and biodegradation processes of FeNTA, likely because they both involve oxidoreduction mechanisms. When the results of the different experiments are compared, the soluble iron, measured by spectrophotometry, was decreasing when microbial cells were present. About 20% of the initial iron was found inside the cells. These results allowed us to propose detailed mechanistic schemes for FeNTA degradation by solar light and by R. rhodochrous .

The transformation of 2-aminobenzothiazole (ABT) was studied under various conditions: (i) a photodegradation process at a λ of >300 nm in the presence of an Fe(III)-nitrilotriacetic acid complex (FeNTA), (ii) a biodegradation process using Rhodococcus rhodochrous OBT18 cells, and (iii) the combined processes (FeNTA plus Rhodococcus rhodochrous in the presence or absence of light). The transformation of ABT in the combined system, with or without light, was much more efficient (99% degradation after 25 h) than in the separated systems (37% photodegradation and 26% biodegradation after 125 h). No direct photolysis of ABT was observed. The main result seen is the strong positive impact of FeNTA on the photodegradation, as expected, and on the biotransformation efficiency of ABT, which was more surprising. This positive impact of FeNTA on the microbial metabolism was dependent on the FeNTA concentration. The use of UV high-performance liquid chromatography, liquid chromatography-electrospray ionization mass spectrometry, and in situ ¹ H nuclear magnetic resonance provided evidence of the intermediary products and thus established transformation pathways of ABT in the different processes. These pathways were identical whether the degradation process was photo- or biotransformation. A new photoproduct was identified (4OH-ABT), corresponding to a hydroxylation reaction on position 4 of the aromatic ring of ABT.

The interactions between microbial and chemical contents of cloud water were investigated. First, we observe that the bulk cloud water solution provides a substantial environment where bacteria can develop significantly. Then, a total number of 60 microbial strains originating from seven distinct samples of cloud water and affiliated to various taxonomic groups were examined for their ability to degrade some of the main atmospheric carboxylic compounds: formate, acetate, lactate, succinate, as well as formaldehyde and methanol. Biodegradation tests show that all these compounds can be transformed when used as single carbonaceous substrates, with activities depending on both the strain and the compound. The highest capacities of biodegradation are observed towards formaldehyde, formate and acetate, which are also the more concentrated compounds typically measured in cloud water. Hence, analyses by ¹H NMR permitted to establish for instance that compounds like pyruvate or fumarate can be produced and released in the media in relation to the transformation of lactate or succinate. In addition, utilization of ¹³C labelled formaldehyde showed that it can be transformed through many metabolic pathways, similar to those induced by photochemistry and leading to the production of formate and/or methanol. These results suggest that microorganisms of cloud water can have various behaviours towards the chemical compounds present in the atmosphere: they can represent either a sink or source for organic carbon, and may have to be considered as actors of cloud chemistry.