Dissipation of pesticides by stream biofilms is influenced by hydrological histories.

To evaluate the effects of hydrological variability on pesticide dissipation capacity by stream biofilms, we conducted a microcosm study. We exposed biofilms to short and frequent droughts (daily frequency), long and less frequent droughts (weekly frequency) and permanently immersed controls, prior to test their capacities to dissipate a cocktail of pesticides composed of tebuconazole, terbuthylazine, imidacloprid, glyphosate and its metabolite aminomethylphosphonic acid. A range of structural and functional descriptors of biofilms (algal and bacterial biomass, extracellular polymeric matrix (EPS) concentration, microbial respiration, phosphorus uptake and community-level physiological profiles) were measured to assess drought effects. In addition, various parameters were measured to characterise the dynamics of pesticide dissipation by biofilms in the different hydrological treatments (% dissipation, peak asymmetry, bioconcentration factor, among others). Results showed higher pesticide dissipation rates in biofilms exposed to short and frequent droughts, despite of their lower biomass and EPS concentration, compared to biofilms in immersed controls or exposed to long and less frequent droughts. High accumulation of hydrophobic pesticides (tebuconazole and terbuthylazine) was measured in biofilms despite the short exposure time (few minutes) in our open-flow microcosm approach. This research demonstrated the stream biofilms capacity to adsorb hydrophobic pesticides even in stressed drought environments.

Vulnerability and tolerance to nickel of periphytic biofilm harvested in summer and winter.

Metals are naturally present in freshwater ecosystems but anthropogenic activities like mining operations represent a long-standing concern. Metals released into aquatic environments may affect microbial communities such as periphytic biofilm, which plays a key role as a primary producer in stream ecosystems. Using two 28-day microcosm studies involving two different photoperiods (light/dark cycle of 16/8 vs 8/16), the present study assessed the effects of four increasing nickel (Ni) concentrations (0-6 µM) on two natural biofilm communities collected at different seasons (summer and winter). The two communities were characterized by different structural profiles and showed significant differences in Ni accumulated content for each treatment. For instance, the biofilm metal content was four times higher in the case of summer biofilm at the highest Ni treatment and after 28 days of exposure. Biomarkers examined targeted both heterotrophic and autotrophic organisms. For heterotrophs, the ß-glucosidase and ß-glucosaminidase showed no marked effects of Ni exposure and were globally similar between the two communities suggesting low toxicity. However, the photosynthetic yield confirmed the toxicity of Ni on autotrophs with maximum inhibition of 81 ± 7% and 60 ± 1% respectively for the summer and winter biofilms. Furthermore, biofilms previously exposed to the highest long-term Ni concentration ([Ni2+] = 6 µM) revealed no acute effects in subsequent toxicity based on the PSII yield, suggesting a tolerance acquisition by the phototrophic community. Taken together, the results suggest that the biofilm response to Ni exposure was dependent of the function considered and that descriptors such as biofilm metal content could be seasonally dependent, information of great importance in a context of biomonitoring.

Temperature and Photoperiod Affect the Sensitivity of Biofilms to Nickel and its Accumulation.

Whereas metal impacts on fluvial communities have been extensively investigated, effects of abiotic parameters on community responses to contaminants are poorly documented. Variations in photoperiod and temperature commonly occur over the course of a season and could affect aquatic biofilm communities and their responses to contaminants. Our objective was to characterize the influence of environmental conditions (photoperiod and temperature) on nickel (Ni) bioaccumulation and toxicity using a laboratory-grown biofilm. Environmental parameters were chosen to represent variations that can occur over the summer season. Biofilms were exposed for 7 days to six dissolved Ni treatments (ranging from 6 to 115 µM) at two temperatures (14 and 20 °C) using two photoperiods (16:8 and 12:12-h light:dark cycle). Under these different scenarios, structural (dry weight biomass and chlorophyll-a) and functional biomarkers (photosynthetic yield and Ni content) were analyzed at four sampling dates, allowing us to evaluate Ni sensitivity of biofilms over time. The results highlight the effects of temperature on Ni accumulation and tolerance of biofilms. Indeed, biofilms exposed at 20 °C accumulated 1.6-4.2-fold higher concentrations of Ni and were characterized by a lower median effect concentration value using photosynthetic yield compared with those exposed at 14 °C. In terms of photoperiod, significantly greater rates of Ni accumulation were observed at the highest tested Ni concentration for biofilms exposed to a 12:12-h compared with a 16:8-h light:dark cycle. Our study demonstrates the influence of temperature on biofilm metabolism and illustrates that environmental factors may influence Ni accumulation response and thus Ni responses of phototrophic biofilms. Environ Toxicol Chem 2022;41:1649-1662. © 2022 SETAC.

Effects of thorium on bacterial, microalgal and micromeiofaunal community structures in a periphytic biofilm.

Few ecotoxicity studies are available on thorium (Th) which hinders the ability to evaluate its ecotoxicological risk. Its release in the environment is often associated with the extraction of rare earth elements and uranium, as well as the field applications of phosphate fertilizers. This study investigates the effects of Th on microbial communities of periphytic biofilms. Ceramic plates were left to colonize for one month in the laboratory with a biofilm sampled from Cap Rouge river (QC, Canada). Plates were randomly placed in channels containing culture media representing three different conditions: a control condition (C0; background Th concentrations of 0.004 ± 0.002 nM), a low Th concentration condition (C1; 0.18 ± 0.09 nM Th) and a moderately high Th condition (C10; 8.7 ± 3.4 nM) for up to 4 weeks. The presence of Th modified the diatom community by changing its taxonomic structure, reducing diversity and increasing cell density. The taxonomic structure of the bacterial community, followed by 16S metabarcoding analysis, was affected with a significant decrease in Pseudanabaena and Shingopyxis genera in the two Th exposed conditions. No direct toxic effect of Th was observed on counted micromeiofauna but the changes in diatom and bacterial communities could explain the higher number of individual diatoms and micromeiofauna observed in Th-exposed conditions. This work shows that low concentrations of Th can modify biofilm structure, which, in turn, could disturb its ecologically key functions.

Thorium Exposure Drives Fatty Acid and Metal Transfer from Biofilms to the Grazer Lymnaea sp.

Aquatic ecotoxicological risks associated with tetravalent metallic elements such as thorium (Th) are still poorly understood. Periphytic biofilm represents an important food source in aquatic environments; thus, such risks could severely affect nutrient and energy cycling in these ecosystems. The present study investigated the potential for Th to change the fatty acid composition of biofilm communities. Bioaccumulation of Th and fatty acids were measured after 4 wk to 2 exposure conditions: a control (C0) and Th exposure (C10). Some major fatty acids such as C16:1n-7 and docosahexaenoic acid C22:6n-3 differed significantly between control and C10 conditions. To determine if Th can be trophically transferred and to investigate the impacts of nutritional quality changes on primary consumers, common pond snails (Lymnaea sp.) were fed for 4 wk with control and Th-exposed biofilm. Thorium appeared to be trophically transferable to the grazers, although we cannot exclude that part of the Th accumulated by the snails may have been taken from the water through release from the biofilms. The composition of major fatty acids observed in the grazers was also significantly affected, notably by a decrease of total polyunsaturated fatty acids. These results indicate that very low Th concentrations can decrease the nutritional quality of organisms at the base of the food chain. Environ Toxicol Chem 2021;40:2220-2228. © 2021 SETAC.

Plastidic Δ6 Fatty-Acid Desaturases with Distinctive Substrate Specificity Regulate the Pool of C18-PUFAs in the Ancestral Picoalga Ostreococcus tauri.

Eukaryotic Δ6-desaturases are microsomal enzymes that balance the synthesis of ω-3 and ω-6 C18-polyunsaturated fatty acids (C18-PUFAs) according to their specificity. In several microalgae, including Ostreococcus tauri, plastidic C18-PUFAs are strictly regulated by environmental cues suggesting an autonomous control of Δ6-desaturation of plastidic PUFAs. Here, we identified two putative front-end Δ6/Δ8-desaturases from O tauri that, together with putative homologs, cluster apart from other characterized Δ6-desaturases. Both were plastid-located and unambiguously displayed a Δ6-desaturation activity when overexpressed in the heterologous hosts Nicotiana benthamiana and Synechocystis sp. PCC6803, as in the native host. Detailed lipid analyses of overexpressing lines unveiled distinctive ω-class specificities, and most interestingly pointed to the importance of the lipid head-group and the nonsubstrate acyl-chain for the desaturase efficiency. One desaturase displayed a broad specificity for plastidic lipids and a preference for ω-3 substrates, while the other was more selective for ω-6 substrates and for lipid classes including phosphatidylglycerol as well as the peculiar 16:4-galactolipid species occurring in the native host. Overexpression of both Δ6-desaturases in O tauri prevented the regulation of C18-PUFA under phosphate deprivation and triggered glycerolipid fatty-acid remodeling, without causing any obvious alteration in growth or photosynthesis. Tracking fatty-acid modifications in eukaryotic hosts further suggested the export of plastidic lipids to extraplastidic compartments.

Metabolome response to anthropogenic contamination on microalgae: a review.

BACKGROUND: Microalgae play a key role in ecosystems and are widely used in ecological status assessment. Research focusing on such organisms is then well developed and essential. Anyway, approaches for a better comprehension of their metabolome's response towards anthropogenic stressors are only emerging. AIM OF REVIEW: This review presents the biochemical responses of various microalgae species towards several contaminants including metals and chemicals as pesticides or industrial compounds. We aim to provide a comprehensive and up-to-date overview of analytical approaches deciphering anthropogenic contaminants impact on microalgae metabolome dynamics, in order to bring out relevant biochemical markers that could be used for risk assessment. KEY SCIENTIFIC CONCEPTS OF REVIEW: Studies to date on ecotoxicological metabolomics on microalgae are highly heterogeneous in both analytical techniques and resulting metabolite identification. There is a real need for studies using complementary approaches to determine biomarkers usable for ecological risk assessment.

Single and mixture effects of pesticides and a degradation product on fluvial biofilms.

The Morcille River located in the Beaujolais vineyard area (Eastern France) is subjected to strong vine-growing pressure leading to the contamination by a range of herbicides and fungicides of the surrounding freshwater environment. Particularly high concentrations of norflurazon, desmethyl norflurazon and tebuconazole were recorded in spring 2010 at the downstream site of the river. Despite their occurrence in rivers, scarce toxicity data are available for these products, in particular in the case of desmethyl norflurazon (main norflurazon degradation product). Furthermore, the toxicity data are generally available only for single compounds and are issued from single species toxicity tests, leading to a lack of ecological relevance. Consequently, this study was undertaken to evaluate the toxic effects of norflurazon, desmethyl norflurazon and tebuconazole singly and in a ternary mixture on fluvial biofilm. Toxicity tests were performed in microplates for 48 h. Photosynthetic endpoints were measured using pulse amplitude-modulated fluorometry; diatom densities and taxonomic composition were determined. After 48 h of exposure, significant effects on optimal quantum yield (F v/F m) for desmethyl norflurazon and mixture were observed.

Development of q-PCR approaches to assess water quality: effects of cadmium on gene expression of the diatom Eolimna minima.

This study was undertaken to develop molecular tools to assess water quality using diatoms as the biological model. Molecular approaches were designed following the development of a rapid and easy RNA extraction method suited to diatoms and the sequencing of genes involved in mitochondrial and photosystem metabolism. Secondly the impact of cadmium was evaluated at the genetic level by q-PCR on 9 genes of interest after exposure of Eolimna minima diatom populations cultured in suspension under controlled laboratory conditions. Their growth kinetics and Cd bioaccumulation were followed. Population growth rates revealed the high impact of Cd at 100 µg/L with total inhibition of growth. These results are linked to the high bioaccumulation values calculated after 14 days of exposure, 57.0±6.3 µg Cd/g dw and 734.1±70 µg Cd/g dw for exposures of 10 and 100 µg Cd/L respectively. Genetic responses revealed the impact of Cd on the mitochondrial metabolism and the chloroplast photosystem of E. minima exposed to 10 and 100 µg Cd/L with induction of cox1, 12S, d1 and psaA after 7 days of exposure for the concentration of 100 µg Cd/L and of nad5, d1 and psaA after 14 days of exposure for both conditions. This is the first reported use of q-PCR for the assessment of toxic pollution on benthic river diatoms. The results obtained presage interesting perspectives, but the techniques developed need to be optimized before the design of new water quality diagnosis tools for use on natural biofilms.

Experimental toxicity and bioaccumulation of cadmium in freshwater periphytic diatoms in relation with biofilm maturity.

A study was undertaken to examine cadmium accumulation in freshwater biofilm, its effects on biofilm development and on diatom community structure in laboratory experimental conditions. A suspension of a biofilm originated from the Riou-Mort River (South West France) was inoculated into three experimental units containing clean glass substrates under laboratory conditions. Settling and already developed biofilms were exposed to a Cd concentration of 100 microg L(-1). Metal accumulation (total and intracellular metal content) in biofilms, dry weight and ash-free dry mass, diatom cell density and diatom community composition were analyzed. Both total and intracellular Cd accumulated by the biofilm throughout the experiment increased with duration of metal exposure. Biofilms in the course of maturation were showed higher Cd content and less effective development than settled biofilms. However diatom communities in younger biofilms exposed to Cd increased their tolerance to Cd by a highly significant development of Nitzschia palea. In contrast, Cd exposure had different effect in installed biofilm and taxonomic composition. These results indicate that mature biofilm may limit Cd accumulation into its architecture and protect diatom communities from the effects of metals.

Seasonal effects of cadmium accumulation in periphytic diatom communities of freshwater biofilms.

The relationships between diatom species and cadmium (Cd) accumulated in biofilms of the Riou-Mort River (SW, France) were studied in July 2004 and March 2005. Biofilms were sampled from artificial substrates immersed along a metallic pollution gradient during 20 days. Dynamics of diatom communities and cadmium accumulation were followed by collecting samples after 4, 7, 14 and 20 days of biofilm colonization. Cd accumulation in biofilms during experiment was significantly higher in Cd polluted station (Joanis) than in reference station (Firmi) for both seasons. Periphytic diatom composition varied between sites and seasons. At Firmi station, seasonal dynamics of diatom communities were stable with the dominance of Cyclotella meneghiniana and Melosira varians in July and Surirellabrebissonnii and Navicula gregaria in March. At Joanis station, diatom communities mainly responded to high levels of metal by a high proportion of small, adnate species. Positive correlations between Eolimna minima, Nitzschia palea, Encyonema minutum, Surirella angusta, and Gomphonema parvulum and cadmium accumulation were observed, indicating that these species are tolerant to high levels of cadmium. On the other hand, negative correlations of C. meneghiniana, N. gregaria, Navicula lanceolata, M. varians and Nitzschia dissipata with cadmium qualify them as sensitive diatom species. Periphytic diatom composition through the presence of specific species highlight metal tolerant indicator diatom groups which will be meaningful for biomonitoring pollution in natural aquatic systems.

SCANNING ELECTRON MICROSCOPY OBSERVATIONS OF DEFORMITIES IN SMALL PENNATE DIATOMS EXPOSED TO HIGH CADMIUM CONCENTRATIONS(1).

Different types of malformations are likely to affect the morphology of diatoms when exposed to particularly unstable environmental conditions, the most easily identifiable being distortion of the whole frustule. In the present study, we investigated, by means of SEM, valve abnormalities induced by high cadmium contamination (100 µg · L(-1) ) in small pennate diatoms. Changes in the shape of Amphora pediculus (Kütz.) Grunow and anomalous sculpturing of the cell wall of many species, such as Encyonema minutum (Hilse) D. G. Mann, Mayamaea agrestris (Hust.) Lange-Bert., Gomphonema parvulum (Kütz.) Kütz., or Eolimna minima (Grunow) Lange-Bert., were observed, which were not, or almost not, noticeable in the LM. With consideration to current knowledge of diatom morphogenesis, metal uptake by the cell would induce, directly or indirectly, damage to many cytoplasmic components (e.g., microtubules, cytoskeleton, Golgi-derived vesicles) involved in the precisely organized silica deposition. This study confirms that many species, whatever their size, are likely to exhibit morphological abnormalities under cadmium stress, and that this indicator may be valuable for the biomonitoring of metal contamination, even if SEM observations are not necessary for routine studies.

Cadmium toxicity and bioaccumulation in freshwater biofilms.

A microcosm study was undertaken to examine the effects of dissolved cadmium at various concentrations (0, 10, and 100 microg . L(-1)) on biofilm accumulation and diatom assemblages. A natural biofilm sampled from the Riou-Mort River (Southwest France) was inoculated into three experimental systems, where biofilm settled on glass slides. Samples collected after 1, 2, 4, and 6 weeks of colonization were analyzed for metal accumulation (total metal content and intracellular metal content in the biofilm), biomass (as measured through dry weight and ash-free dry matter), and quantitative as well as qualitative analysis of diatom assemblages. There was a positive correlation between cadmium accumulation and dissolved cadmium concentrations and duration of exposure: a linear relationship was found between concentration factors (CFs) of growing biofilms and time (CFs/day = 0.25 and 0.38 under contaminations of 10 and 100 microgCd . L(-1), respectively). Biofilm settlement, more than photosynthetic activity, was affected by high cadmium concentrations: we observed for all stages of settlement a drastic and significant (p < 0.05) reduction in biofilm biomass and in diatom densities in the highest cadmium contamination, compared to control and low cadmium concentration units.