Catarina R. Marques

Toxicity evaluation of three pesticides on non-target aquatic and soil organisms: commercial formulation versus active ingredient

The Ecological Risk Assessment of pesticides requires data regarding their toxicity to aquatic and terrestrial non-target species. Such requirements concern active ingredient(s), generally not considering the noxious potential of commercial formulations. This work intends to contribute with novel information on the effects of short-term exposures to two herbicides, with different modes of action (Spasor®, Stam Novel Flo 480®), and an insecticide (Lannate®), as well as to corresponding active ingredients (Glyphosate, Propanil and Methomyl, respectively). The microalga Pseudokirchneriella subcapitata (growth inhibition), the cladoceran Daphnia magna (immobilisation), and the earthworm Eisenia andrei (avoidance behaviour) were used as test species. Both herbicides were innocuous to all test organisms at environmentally realistic concentrations, except for Stam and Propanil (highly toxic for Pseudokirchneriella; moderately toxic to Daphnia). Lannate and Methomyl were highly toxic to Daphnia and caused Eisenia to significantly avoid the spiked soil at realistic application rates. The toxicity of formulations either overestimated (e.g. Stam/Propanil for P. subcapitata) or underestimated (e.g. Stam/Propanil for D. magna) that of the active ingredient.

A whole sample toxicity assessment to evaluate the sub-lethal toxicity of water and sediment elutriates from a lake exposed to diffuse pollution

The impact of diffuse pollution in aquatic systems is of great concern due to the difficult to measure and regulate it. As part of an ecological risk assessment (ERA), this study aims to use a whole sample toxicity assessment to evaluate the toxicity of water and sediment from Lake Vela, a lake that has been exposed to diffuse pollution. In this way, standard (algae: Pseudokirchneriella subcapitata; cladoceran: Daphnia magna) and local species (algae: Aphanizomenon flos‐aquae; cladoceran: Daphnia longispina) were exposed to surface water, and sediment elutriates were collected seasonally from two sites at Lake Vela: one near the east bank (ES), surrounded by agricultural lands; and the other near the west bank (WS), surrounded by a forest. The results confirmed the seasonal contamination of both environmental compartments by pesticides, including organochlorine pesticides, and the presence of high concentrations of nutrients. Although both sites were contaminated, higher levels of pesticides and nutrients were detected in ES, particularly in the sediments. Bioassays showed that water samples (100% concentration) collected in summer and autumn significantly affected the growth rate of P. subcapitata, which could be attributed to the presence of pesticides. Likewise, they revealed an apparent toxicity of elutriates for P. subcapitata and for both daphnids, in summer and autumn. In fact, although pesticides were not detected in elutriates, high levels of un‐ionized ammonia were recorded, which is considered highly toxic to aquatic life. By comparing the several species, P. subcapitata was revealed to be the most sensitive one, followed by the daphnids, and then by A. flos‐aquae. Results obtained in this study underlined the importance of whole samples toxicity assessment for characterizing the ecological effects of complex mixtures from diffuse inputs, in the ERA processes.

Unraveling the interactive effects of climate change and oil contamination on laboratory-simulated estuarine benthic communities.

There is growing concern that modifications to the global environment such as ocean acidification and increased ultraviolet radiation may interact with anthropogenic pollutants to adversely affect the future marine environment. Despite this, little is known about the nature of the potential risks posed by such interactions. Here, we performed a multifactorial microcosm experiment to assess the impact of ocean acidification, ultraviolet B (UV-B) radiation and oil hydrocarbon contamination on sediment chemistry, the microbial community (composition and function) and biochemical marker response of selected indicator species. We found that increased ocean acidification and oil contamination in the absence of UV-B will significantly alter bacterial composition by, among other things, greatly reducing the relative abundance of Desulfobacterales, known to be important oil hydrocarbon degraders. Along with changes in bacterial composition, we identified concomitant shifts in the composition of oil hydrocarbons in the sediment and an increase in oxidative stress effects on our indicator species. Interestingly, our study identifies UV-B as a critical component in the interaction between these factors, as its presence alleviates harmful effects caused by the combination of reduced pH and oil pollution. The model system used here shows that the interactive effect of reduced pH and oil contamination can adversely affect the structure and functioning of sediment benthic communities, with the potential to exacerbate the toxicity of oil hydrocarbons in marine ecosystems.

Toxicity evaluation of natural samples from the vicinity of rice fields using two trophic levels.

An ecotoxicological screening of environmental samples collected in the vicinity of rice fields followed a combination of physical and chemical measurements and chronic bioassays with two freshwater trophic levels (microalgae: Pseudokirchneriella subcapitata and Chlorella vulgaris; daphnids: Daphnia longispina and Daphnia magna). As so, water and sediment/soil elutriate samples were obtained from three sites: (1) in a canal reach crossing a protected wetland upstream, (2) in a canal reach surrounded by rice fields and (3) in a rice paddy. The sampling was performed before and during the rice culture. During the rice cropping, the whole system quality decreased comparatively to the situation before that period (e.g. nutrient overload, the presence of pesticides in elutriates from sites L2 and L3). This was reinforced by a significant inhibition of both microalgae growth, especially under elutriates. Contrary, the life-history traits of daphnids were significantly stimulated with increasing concentrations of water and elutriates, for both sampling periods.

Impact of organic nano-vesicles in soil: The case of sodium dodecyl sulphate/didodecyl dimethylammonium bromide.

Aiming at contributing new insights into the effects of nanomaterials (NMs) in the terrestrial ecosystem, this study evaluated the impacts of organic nano-vesicles of sodium dodecyl sulphate/didodecyl dimethylammonium bromide (SDS/DDAB) on the emergence and growth of plant seeds, and on the avoidance and reproduction of soil invertebrates. For this purpose several ecotoxicological assays were performed with different test species (terrestrial plants: Zea mays, Avena sativa, Brassica oleracea and Lycopersicon esculentum; soil invertebrates: Eisenia andrei and Folsomia candida). A wide range of SDS/DDAB concentrations were tested, following standard protocols, and using the standard OECD soil as a test substrate (5% of organic matter). The aqueous suspensions of SDS/DDAB, used to spike the soils, were characterised by light scattering techniques for hydrodynamic size of the vesicles, aggregation index, polydispersity index, zeta potential and surface charge. The SDS/DDAB concentrations in the test soil were analysed by HPLC-UV at the end of the assays. Invertebrate species were revealed to be sensitive to nano-SDS/DDAB upon immediate exposure to freshly spiked soils. However, the degradation of SDS/DDAB nano-vesicles in the soil with time prevented the occurrence of significant reproduction effects on soil invertebrates. Plants were not particularly sensitive to SDS/DDAB, except B. oleracea (at concentrations above 375 mg kg(-1)dw). The results gathered in this study allowed a preliminary determination of a risk limit to nano-SDS/DDAB. The low toxicity of SDS/DDAB nano-vesicles could be explained by its high and fast degradation in the soil. The soil microbial community could have an important role in the fate of this NM, thus it is of remarkable importance to improve this risk limit by taking into account specific data addressing this community.

Contribution for the Derivation of a Soil Screening Value (SSV) for Uranium, Using a Natural Reference Soil

In order to regulate the management of contaminated land, many countries have been deriving soil screening values (SSV). However, the ecotoxicological data available for uranium is still insufficient and incapable to generate SSVs for European soils. In this sense, and so as to make up for this shortcoming, a battery of ecotoxicological assays focusing on soil functions and organisms, and a wide range of endpoints was carried out, using a natural soil artificially spiked with uranium. In terrestrial ecotoxicology, it is widely recognized that soils have different properties that can influence the bioavailability and the toxicity of chemicals. In this context, SSVs derived for artificial soils or for other types of natural soils, may lead to unfeasible environmental risk assessment. Hence, the use of natural regional representative soils is of great importance in the derivation of SSVs. A Portuguese natural reference soil PTRS1, from a granitic region, was thereby applied as test substrate. This study allowed the determination of NOEC, LOEC, EC20 and EC50 values for uranium. Dehydrogenase and urease enzymes displayed the lowest values (34.9 and <134.5 mg U Kg, respectively). Eisenia andrei and Enchytraeus crypticus revealed to be more sensitive to uranium than Folsomia candida. EC50 values of 631.00, 518.65 and 851.64 mg U Kg were recorded for the three species, respectively. Concerning plants, only Lactuca sativa was affected by U at concentrations up to 1000 mg U kg1. The outcomes of the study may in part be constrained by physical and chemical characteristics of soils, hence contributing to the discrepancy between the toxicity data generated in this study and that available in the literature. Following the assessment factor method, a predicted no effect concentration (PNEC) value of 15.5 mg kg−1 dw was obtained for U. This PNEC value is proposed as a SSV for soils similar to the PTRS1.

Toxicity screening of soils from different mine areas—A contribution to track the sensitivity and variability of Arthrobacter globiformis assay

This study used the Arthrobacter globiformis solid-contact test for assessing the quality of soils collected in areas subjected to past and present mine activities in Europe (uranium mine, Portugal) and North Africa (phosphogypsum pile, Tunisia; iron mine, Morocco). As to discriminate the influence of soils natural variability from the effect of contaminants, toxicity thresholds were derived for this test, based on the dataset of each study area. Furthermore, the test sensitivity and variability was also evaluated. As a result, soils that inhibited A. globiformis dehydrogenase activity above 45% or 50% relatively to the control, were considered to be toxic. Despite the soil metal content determined, the properties of soils seemed to influence dehydrogenase activity. Overall, the contact test provided a coherent outcome comparing to other more time-consuming and effort-demanding ecotoxicological assays. Our results strengthened the feasibility and ecological relevance of this assay, which variability was quite reduced hence suggesting its potential integration within the test battery of tier 1 of soil risk assessment schemes.

In situ aquatic bioassessment of pesticides applied on rice fields using a microalga and daphnids

This study assessed the effects of episodic contamination on a drainage canal adjacent to an area of intensive rice production (Coimbra, Portugal). Four monitoring periods were considered [i) before herbicide application (day-14), ii) at the first application day (day 0), iii) 3 or 5 and iv) 6days after]. Each one consisted in three complementary evaluation lines: a) physico-chemical analyses, b) whole effluent toxicity (WET) assays with Pseudokirchneriella subcapitata, c) in situ bioassays to assess microalgae (P. subcapitata) growth, and the feeding rate and survival of Daphnia longispina and Daphnia magna. Study sites were located upstream, in a protected wetland (L1), and downstream, in the vicinity of rice fields (L2). Along with the application of agrochemicals, there was a general decrease of the water quality, especially in L2, due to nutrient and herbicide inputs. Herbicide peaks (on days 0, 5 and 6) in L2 water samples were recorded concomitantly or immediately after their application. Regarding the in situ bioassessment, the algae growth decrease from day 0 onwards in L1, whilst in L2 its inhibition was generally coherent with the decline of the water quality. Apparently, WET tests indicated that the limitation of nutrients could be affecting algae growth in L1, however, conclusions should be cautious. The feeding depression of daphnids occurred on days 0 and 5 for D. longispina and only on day 0 for D. magna, while significant reductions on survival were restricted to day 0 for both species. The impairments occurring on day 0 were linked to a potential increased toxicity driven by the ingestion of particle-bound herbicides and suspended particles. The feeding rate of daphnids provided an earlier indication of toxic impairments, though it is prompted the use of complementary endpoints and trophic levels in order to understand the cumulative effects due to various herbicide pulses.

Ecotoxicological effects of Mikado® and Viper® on algae and daphnids

The toxicity of single and combined formulated herbicides (Mikado® and Viper®) was assessed on several endpoints in species from two trophic levels: algae growth—Pseudokirchneriella subcapitata and Chlorella vulgaris—immobilization and life‐history traits (only for single compound toxicity) of daphnids—Daphnia longispina and Daphnia magna. Viper was the most toxic formulated herbicide. It was hypothesized that the toxicity of both formulated herbicides could have been enhanced by adjuvants, especially for Viper. In most cases, the sublethal endpoints were the most sensitive and affected by both formulations, comparatively to their acute effects. Concentration addition (CA) and independent action (IA) models provided an accurate description of Mikado and Viper joint action on algae growth and immobilization of daphnids, although significant deviations were always detected. A low‐dose antagonism and high‐dose synergism were identified for P. subcapitata, whereas C. vulgaris response deviated antagonistically from CA and synergistically from IA. For both daphnids, however, synergistic effects were observed for higher mixture concentrations. Under a regulatory standpoint, CA provided the most conservative estimation either because the mixture effects were overestimated or less subestimated than IA. Overall, the great sensitivity differences observed within species did not allow the conclusion that one trophic level was more tolerant than the other. Instead, P. subcapitata was always the most sensitive species to both herbicide formulations, followed by D. longispina, while D. magna and C. vulgaris were the most tolerant species. On a whole, further studies are needed toward a comprehensive understanding of herbicides mode of action, their effects at lower biological‐level endpoints, and under different mixture designs.

Bio-rescue of marine environments: On the track of microbially-based metal/metalloid remediation.

The recent awareness of the huge relevance of marine resources and ecological services is driving regulatory demands for their protection from overwhelming contaminants, such as metals/metalloids. These contaminants enter and accumulate in different marine niches, hence deeply compromising their quality and integrity. Bioremediation has been flourishing to counteract metal/metalloid impacts, since it provides cost-effective and sustainable options by relying on ecology-based technologies. The potential of marine microbes for metal/metalloid bioremediation is the core of many studies, due to their high plasticity to overcome successive environmental hurdles. However, any thorough review on the advances of metal/metalloid bioremediation in marine environments was so far unveiled. This review is designed to (i) outline the characteristics and potential of marine microbes for metal/metalloid bioremediation, (ii) describe the underlying pathways of resistance and detoxification, as well as useful methodologies for their characterization, (iii) identify major bottlenecks on metal/metalloid bioremediation with marine microbes, (iv) present alternative strategies based on microbial consortia and engineered microbes for enhanced bioremediation, and (v) propose key research avenues to keep pace with a changing society, science and economy in a sustainable manner.