Analytical methodologies for arsenic speciation in macroalgae: a critical review

Marine and estuarine macroalgae are increasingly used for direct consumption, due to its high content of essential nutrients. However, the toxicity of chemical forms of arsenic (As) present in macroalgae are becoming of great concern and an important research topic. This review will discuss the sophisticated analytical methodologies used for arsenic speciation, as well the investigation of cheaper and fit for purpose methods able to inform about the safety of macroalgae consumption. Furthermore, we will also discuss relevant speciation studies performed by various authors highlighting the methodologies and respective figures of merit for all components of As in macroalgae, not only inorganic As (iAs). Focusing on the steps prior to speciation, such as sample extraction and clean-up, it will be stressed the need to develop certified reference materials (CRMs) in order to assist in the validation of analytical methodologies for As speciation in macroalgae.publishe

Evaluation of multivariate biomarker indexes application in ecotoxicity tests with marine diatoms exposed to emerging contaminants

Worldwide anthropogenic activities result in the production and release of potentially damaging toxic pollutants into ecosystems, thereby jeopardizing their health and continuity. Research studies and biomonitoring programs attend to this emerging problematic by applying and developing statistically relevant indexes that integrate complex biomarker response data to provide a holistic approach, reflecting toxically induced alterations at the organism or population level. Ultimately, indexes allow simple result communications, enhancing policy makers understanding, and contributing to better resource and environmental managing policies. In this study three indexes, the integrated biomarker response index (IBR), the bioeffects assessment index (BAI) and principal components analysis (PCA), were evaluated for their sensitivity in revealing toxically induced stress patterns in cells of the diatom Phaeodactylum tricornutum under contaminant exposure. The set of biomarkers selected for index construction comprised the anti-oxidant enzymes APX, CAT and SOD, and the lipid peroxidation marker TBARS. Several significant correlations with the applied concentration gradients were noticed for all indexes, although IBR excelled for its reliability in delivering statistically significant dose-response patterns for four out of the five tested compounds.info:eu-repo/semantics/publishedVersio

Effects on biomarkers in stress ecology studies. Well, so what? What now?

The authors would like to thank Fundação para a Ciência e a Tecnologia (FCT) for the support through funding of MARE (UIDB/04292/2020 and UIDP/04292/2020), ARNET—Aquatic Research Network Associated Laboratory (LA/P/0069/2020), MARINE INVADERS (PTDC/BIA-CBI/31144/2017), BLUESHARKER (PTDC/BIA-CBI/29136/2017), and OPTOX (PTDC/CTA-AMB/30056/2017). B. Duarte and V. F. Fonseca were supported by researcher contracts (CEEC-IND/00511/2017 and 2021.00244.CEECIND, respectively).1. Biomarkers in Stress Ecology - From the Gene to Population Effects assessed at higher levels of biological organization (populations and communities) are the consequence of the sum of effects on individuals, which usually result from impacts at cellular and molecular levels. Given this rationale, these lower levels of biological organization are more responsive at an early stage, making them potential resources that can be used as early warning endpoints to address environmental stress. In this way, the information concerning effects at the molecular level of biological organization (e.g., transcripts, proteins, or metabolites) allows for an early assessment of future ecosystem problems, which may eventually enable a timely intervention before the impacts become visible and irreversible. However, despite providing an early warning and a better understanding of the toxicity mechanisms, enabling the protection of biological integrity, the most significant setback is that these endpoints may fail to foresee later impacts on the environment due to the ecosystem resilience or a weak link to the effects in the following level of biological organization, making these tools simply too conservative for stakeholders’ interests. Hence, an approach targeting lower levels of biological organization will greatly benefit from addressing potential effects at higher levels. This can be achieved by establishing a link in biological organization, where the effects assessed at the lower end of biological organization are linked with the high probability of causing an effect at the other end, inducing changes in populations and communities, and eventually altering ecosystems in the future.info:eu-repo/semantics/publishedVersio

A new approach for routine quantification of microplastics using Nile Red and automated software (MP-VAT)

Microplastics are widespread contaminants in the environment. However, most identification protocols rely on long and subjective visual counting, which could be improved using staining dyes. Thus, the objective of this work is to identify the best staining dye protocol and create an objective and quick automated counting software for microplastics. Tests were conducted to identify the most appropriate of eight staining dye solutions and of six wavelengths for virgin and weathered synthetic polymers, textile fibers, natural organic matter and filters. Nile Red produced the best results (without interfering in infrared spectra) rendering microplastics fluorescent at 254 nm, but with limited number of fluorescent polymers, and at 470 nm (with orange filter), with fluorescence of plastics as well as natural organic matter (requiring a digestion step). Next, a script was developed in ImageJ for the automatic quantification and characterization in shape (fiber, fragment, particle) and size of fluorescent microplastics, the Microplastics Visual Analysis Tool (MP-VAT). MP-VAT was evaluated, producing recovery rates in the range of 89.0-111.1% in spiked filters under 470 nm. Furthermore, this package is accompanied by a script that sets a scale from a known filter diameter, MP-SCALE, and a script that allows user threshold setting, MP-ACT.publishe

Marked changes in diversity and relative activity of picoeukaryotes with depth in the world ocean

Microbial eukaryotes are key components of the ocean plankton. Yet, our understanding of their community composition and activity in different water layers of the ocean is limited, particularly for picoeukaryotes (0.2–3 µm cell size). Here, we examined the picoeukaryotic communities inhabiting different vertical zones of the tropical and subtropical global ocean: surface, deep chlorophyll maximum, mesopelagic (including the deep scattering layer and oxygen minimum zones), and bathypelagic. Communities were analysed by high-tthroughput sequencing of the 18S rRNA gene (V4 region) as represented by DNA (community structure) and RNA (metabolism), followed by delineation of Operational Taxonomic Units (OTUs) at 99% similarity. We found a stratification of the picoeukaryotic communities along the water column, with assemblages corresponding to the sunlit and dark ocean. Specific taxonomic groups either increased (e.g., Chrysophyceae or Bicosoecida) or decreased (e.g., Dinoflagellata or MAST-3) in abundance with depth. We used the rRNA:rDNA ratio of each OTU as a proxy of metabolic activity. The highest relative activity was found in the mesopelagic layer for most taxonomic groups, and the lowest in the bathypelagic. Altogether, we characterize the change in community structure and metabolic activity of picoeukaryotes with depth in the global ocean, suggesting a hotspot of activity in the mesopelagic

Comparison of loop-mediated isothermal amplification, polymerase chain reaction, and selective isolation assays for detection of Xanthomonas albilineans from sugarcane

A loop-mediated isothermal amplification (LAMP) assay was developed and compared to polymerase chain reaction (PCR), nested PCR, and selective isolation assays for detection of Xanthomonas albilineans, the causal agent of sugarcane leaf scald. The pathogen was isolated on selective medium from 44 out of 45 (98%) samples taken from symptomatic stalks, and from 44 out of 70 (63%) samples from asymptomatic stalks that were collected from plots with symptomatic stalks. Forty-two (93%), 41 (91%), and 42 (93%) symptomatic samples tested positive by LAMP, PCR and nested PCR, respectively. The pathogen was detected in 19 (27%), 8 (11%), and 25 (36%) of the 70 asymptomatic samples by LAMP, PCR and nested PCR, respectively. Symptomatic stalks were mainly, but not always, associated with high populations of the pathogen (107–109 CFU/ml), and asymptomatic stalks with low populations (<103 CFU/ml) or no bacteria. Although our LAMP and nested PCR methods detected 10 CFU/ml of X. albilineans in suspensions prepared with pure culture, they sometimes failed to detect the pathogen in samples with low pathogen populations. Isolation on selective medium along with another method should therefore be used for detection of the pathogen in asymptomatic stalks, especially in quarantine programs

Dimensioning of a multibeam coherent photonic beamformer fed by a phased array antenna.

The design and dimensioning of a photonic-aided payload for a multi-beam high-throughput communications satellite is a complex problem in which the antenna, RF and photonic subsystems must be considered as a whole for achieving best performance with lowest mass and power consumption. In this paper, we propose and dimension the receiving stage of a communications satellite comprising a phased array antenna (PAA) feeding a multibeam photonic beamforming system (PBS). The PBS uses a single wavelength and resorts to heterodyne detection such that the retrieved beams are frequency downconverted. End-to-end system modeling shows that the complexity of the PAA and PBS can be traded-off for signal-to-noise ratio (SNR) or power consumption without compromising the beam width. The dimensioning of a realistic scenario is presented, showing that an SNR and beam crosstalk on the order of 20 dB are achievable with a total power consumption below 1 kW for a typical number of 100 antenna elements (AEs)

Fluoxetine Arrests Growth of the Model Diatom Phaeodactylum tricornutum by Increasing Oxidative Stress and Altering Energetic and Lipid Metabolism

Pharmaceutical residues impose a new and emerging threat to aquatic environments and its biota. One of the most commonly prescribed pharmaceuticals is the antidepressant fluoxetine, a selective serotonin re-uptake inhibitor that has been frequently detected, in concentrations up to 40 μg L–1, in aquatic ecosystems. The present study aims to investigate the ecotoxicity of fluoxetine at environmentally relevant concentrations (0.3, 0.6, 20, 40, and 80 μg L–1) on cell energy and lipid metabolism, as well as oxidative stress biomarkers in the model diatom Phaeodactylum tricornutum. Exposure to higher concentrations of fluoxetine negatively affected cell density and photosynthesis through a decrease in the active PSII reaction centers. Stress response mechanisms, like β-carotene (β-car) production and antioxidant enzymes [superoxide dismutase (SOD) and ascorbate peroxidase (APX)] up-regulation were triggered, likely as a positive feedback mechanism toward formation of fluoxetine-induced reactive oxygen species. Lipid peroxidation products increased greatly at the highest fluoxetine concentration whereas no variation in the relative amounts of long chain polyunsaturated fatty acids (LC-PUFAs) was observed. However, monogalactosyldiacylglycerol-characteristic fatty acids such as C16:2 and C16:3 increased, suggesting an interaction between light harvesting pigments, lipid environment, and photosynthesis stabilization. Using a canonical multivariate analysis, it was possible to evaluate the efficiency of the application of bio-optical and biochemical techniques as potential fluoxetine exposure biomarkers in P. tricornutum. An overall classification efficiency to the different levels of fluoxetine exposure of 61.1 and 88.9% were obtained for bio-optical and fatty acids profiles, respectively, with different resolution degrees highlighting these parameters as potential efficient biomarkers. Additionally, the negative impact of this pharmaceutical molecule on the primary productivity is also evident alongside with an increase in respiratory oxygen consumption. From the ecological point of view, reduction in diatom biomass due to continued exposure to fluoxetine may severely impact estuarine and coastal trophic webs, by both a reduction in oxygen primary productivity and reduced availability of key fatty acids to the dependent heterotrophic upper levels.info:eu-repo/semantics/publishedVersio

Effects of Propranolol on Growth, Lipids and Energy Metabolism and Oxidative Stress Response of Phaeodactylum tricornutum

Present demographic trends suggest a rise in the contributions of human pharmaceuticals into coastal ecosystems, underpinning an increasing demand to evaluate the ecotoxicological effects and implications of drug residues in marine risk assessments. Propranolol, a non-selective β-adrenoceptor blocker, is used worldwide to treat high blood pressure conditions and other related cardiovascular conditions. Although diatoms lack β-adrenoceptors, this microalgal group presents receptor-like kinases and proteins with a functional analogy to the animal receptors and that can be targeted by propranolol. In the present work, the authors evaluated the effect of this non-selective β-adrenoceptor blocker in diatom cells using P. tricornutum as a model organism, to evaluate the potential effect of this compound in cell physiology (growth, lipids and energy metabolism and oxidative stress) and its potential relevance for marine ecosystems. Propranolol exposure leads to a significant reduction in diatom cell growth, more evident in the highest concentrations tested. This is likely due to the observed impairment of the main primary photochemistry processes and the enhancement of the mitochondrial respiratory activity. More specifically, propranolol decreased the energy transduction from photosystem II (PSII) to the electron transport chain, leading to an increase in oxidative stress levels. Cells exposed to propranolol also exhibited high-dissipated energy flux, indicating that this excessive energy is efficiently diverted, to some extent, from the photosystems, acting to prevent irreversible photoinhibition. As energy production is impaired at the PSII donor side, preventing energy production through the electron transport chain, diatoms appear to be consuming storage lipids as an energy backup system, to maintain essential cellular functions. This consumption will be attained by an increase in respiratory activity. Considering the primary oxygen production and consumption pathways, propranolol showed a significant reduction of the autotrophic O2 production and an increase in the heterotrophic mitochondrial respiration. Both mechanisms can have negative effects on marine trophic webs, due to a decrease in the energetic input from marine primary producers and a simultaneous oxygen production decrease for heterotrophic species. In ecotoxicological terms, bio-optical and fatty acid data appear as highly efficient tools for ecotoxicity assessment, with an overall high degree of classification when these traits are used to build a toxicological profile, instead of individually assessed.info:eu-repo/semantics/publishedVersio

Fluoxetine induces photochemistry-derived oxidative stress on Ulva lactuca

Emerging pollutants impose a high degree of stress on marine ecosystems, compromising valuable resources, the planet and human health. Pharmaceutical residues often reach marine ecosystems, and their input is directly related to human activities. Fluoxetine is an antidepressant, and one of the most prescribed selective serotonin reuptake inhibitors globally and has been detected in aquatic ecosystems in concentrations up to 40 μg L−1 . The present study aims to evaluate the impact of fluoxetine ecotoxicity on the photochemistry, energy metabolism and enzyme activity of Ulva lactuca exposed to environmentally relevant concentrations (0.3, 0.6, 20, 40, and 80 μg L−1 ). Exogenous fluoxetine exposure induced negative impacts on U. lactuca photochemistry, namely on photosystem II antennae grouping and energy fluxes. These impacts included increased oxidative stress and elevated enzymatic activity of ascorbate peroxidase and glutathione reductase. Lipid content increased and the altered levels of key fatty acids such as hexadecadienoic (C16:2) and linoleic (C18:2) acids revealed strong correlations with fluoxetine concentrations tested. Multivariate analyses reinforced the oxidative stress and chlorophyll a fluorescence-derived traits as efficient biomarkers for future toxicology studies.info:eu-repo/semantics/publishedVersio