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Chlorpyrifos, an organophosphate insecticide widely used to control agricultural pests, poses a significant environmental threat due to its toxicity and persistence in soil and water. Our work aimed to evaluate the acute (survival) and chronic (regeneration, locomotion, and reproduction) toxicity of chlorpyrifos to the non-target freshwater planarian Girardia tigrina . The 48 h lethal concentration (LC 50 ) of the commercial formulation, containing 480 g L -1 of chlorpyrifos, the active ingredient, was determined to be 622.8 µg a.i. L -1 for planarians. Sublethal effects were translated into a significant reduction in locomotion and delayed head regeneration (lowest observed effect concentration-LOEC = 3.88 µg a.i. L -1 ). Additionally, chlorpyrifos exposure did not affect planarian fecundity or fertility. Overall, this study demonstrates the potential of chlorpyrifos-based insecticides to harm natural populations of freshwater planarians at environmentally relevant concentrations. The observed toxicity emphasizes the need for stricter regulations and careful management of chlorpyrifos usage to mitigate its deleterious effects on aquatic ecosystems. By understanding the specific impacts on non-target organisms like G. tigrina , we can make more informed suggestions regarding the usage and regulation of organophosphate insecticides, ultimately promoting sustainable agricultural practices and environmental conservation.

Every year, millions of people suffer some form of illness associated with the consumption of contaminated food. Escherichia coli (E. coli), found in the intestines of humans and other animals, is commonly associated with various diseases, due to the existence of pathogenic strains. Strict monitoring of food products for human consumption is essential to ensure public health, but traditional cell culture-based methods are associated with long waiting times and high costs. New approaches must be developed to achieve cheap, fast, and on-site monitoring. Thus, in this work, we developed optical fiber sensors based on surface plasmon resonance. Gold and cysteamine-coated fibers were functionalized with anti-E. coli antibody and tested using E. coli suspensions with concentrations ranging from 1 cell/mL to 10 5 cells/mL. An average logarithmic sensitivity of 0.21 ± 0.01 nm/log(cells/mL) was obtained for three independent assays. An additional assay revealed that including molybdenum disulfide resulted in an increase of approximately 50% in sensitivity. Specificity and selectivity were also evaluated, and the sensors were used to analyze contaminated water samples, which verified their promising applicability in the aquaculture field.

Disposable facemasks represent a new form of environmental contamination worldwide. This study aimed at addressing the abundance of facemasks in an overlooked natural environment with high ecological and economic value - the wetlands (Ria de Aveiro, Portugal, as study case), evaluating their potential biodegradation using naturally occurring fungi and assessing the potential ecotoxicity of released microfibres on local bivalves. All masks collected within 6500 m 2 area of Aveiro wetland were 100 % disposable ones (PP-based, confirmed by Fourier transform infrared spectroscopy - FTIR) with an initial abundance of 0.0023 items/m 2 in Sept. 2021, which was reduced by ∼40 % in Apr. 2022 and ∼87 % in Sept. 2022, as a reflection of the government policies. Analysis of the carbonyl index (0.03 to 1.79) underlined their state of degradation, primarily due to sun exposure during low tides. In laboratory conditions, 1 mm 2 microplastics obtained from new disposable facemasks were prone to biodegradation by Penicillium brevicompactum and Zalerion maritimum inferred from microplastics mass loss (∼22 to -26 % and ∼40 to 50 %, respectively) and FTIR spectra (particularly in the hydroxyl and carbonyl groups). In addition, microfibres released from facemasks induced sublethal effects on the clam, Venerupis corrugata, mostly in their UV-aged form when compared to pristine ones, characterised by a decrease in cellular energy allocation (CEA) and an increase in aerobic energy metabolism (ETS). Concomitantly, clams exposed to 1250 items/L of UV-aged microplastics (similar to field-reported concentrations) expressed greater clearance capacity, indicating a need to compensate for the potential energy unbalance. This study provides the first baseline monitoring of facemasks in wetlands while bringing new evidence on their biodegradation and ecotoxicity, considering environmentally relevant conditions and keystone organisms in such environments. Such studies require scientific attention for rapid regulatory action against this emerging and persistent pollutant, also targeting remediation and mitigation strategies considering these items under pandemic scenarios., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Microplastic pollution, global warming, and invasive species are known threats to marine biota, but the impact of their simultaneous exposure is still not well understood. This study investigated whether the toxic effects posed by the invasive red seaweed Asparagopsis armata exudate (2%) to the mussel Mytilus galloprovincialis are amplified by a 96 h exposure to increased temperature (24 °C) and polyethylene microplastics (PE-MPs, 1 mg/L). Biochemical (neurotoxicity, energy metabolism, oxidative stress, and damage) and physiological (byssal thread production) responses were evaluated. The number of produced byssus greatly decreased under concomitant exposure to all stressors. The antioxidant defences were depleted in the gills of mussels exposed to temperature rises and PE-MPs, regardless of exudate exposure, preventing oxidative damage. Moreover, the heat shock protein content tended to decrease in all treatments relative to the control. The increased total glutathione in the mussels' digestive gland exposed to 24 °C, exudate, and PE-MPs avoided oxidative damage. Neurotoxicity was observed in the same treatment. In contrast, the energy metabolism remained unaltered. In conclusion, depending on the endpoint, simultaneous exposure to A. armata exudate, PE-MPs, and warming does not necessarily mean an amplification of their single effects. Studies focusing on the impact of multiple stressors are imperative to better understand the underlying mechanisms of this chronic exposure.

The greater white-toothed shrew Crocidura russula has been used as a sentinel species for estimating environmental risks to human populations. Previous studies in mining areas have focused on the liver of shrews as the primary target of physiological and metabolic changes due to heavy metal pollution. However, populations persist even when detoxification by the liver seems to be compromised and damage is observed. These pollutant-adapted individuals inhabiting contaminated sites may exhibit altered biochemical parameters that confer increased tolerance in various tissues other than the liver. The skeletal muscle tissue of C. russula might be an alternative tissue that allows the survival of organisms inhabiting historically polluted sites due to the detoxification of redistributed metals. Organisms from two heavy metal mine populations and one population derived from an unpolluted site were used to determine the detoxification activities, antioxidant capacity, and oxidative damage, as well as cellular energy allocation parameters and acetylcholinesterase activity (a biomarker of neurotoxicity). Muscle biomarkers differ between shrews from polluted sites and shrews from the unpolluted location, with the mine animals showing: (1) a decreased energy consumption concomitant with increased energy reserves and total available energy; (2) reduced cholinergic activity, suggesting an impairment of neurotransmission at the neuromuscular junction; (3) an overall decrease in detoxification capacity and enzymatic antioxidant response and a higher level of lipid damage. Also, some of these markers differed between females and males. These changes may have resulted from a decreased detoxifying capacity of the liver and could potentially bring about significant ecological effects for this highly active species. Heavy metal pollution induced physiological changes in Crocidura russula showing that skeletal muscle may serve as a backup sink organ allowing rapid species adaptation and evolution., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Bivalve mollusks represent a nutritious source with a low environmental impact; as a result, they are one of the most attractive aquaculture options. Advances in microencapsulation technology offer great potential to face key bivalve nutrition problems, and an alga-based microencapsulated diet can turn enriched bivalves into potential functional foods. The central goal of this study was the evaluation of food intake as a function of particle size and microalga content following the supply of four microencapsulated diets, incorporating as core material Nannochloropsis sp. or Tetraselmis sp. in 20 or 40 µm diameter pellets (diets N20, T20, N40, and T40, respectively) in five bivalve species ( Magallana gigas , Solen marginatus , Ruditapes decussatus , Ruditapes philippinarum , and Cerastoderma edule ). Overall, all tested diets were easily ingested, although food intake was higher for N20 (except for the S. marginatus , which showed a higher rate for the diet T40). Concerning a size-related analysis, C. edule and S. marginatus favored, respectively, smaller and bigger pellet-sized diets, with no signs of selectivity for microalga species. The diet T20 was the lesser ingested, except for C. edule . This knowledge enables a better selection of feed with appropriate and species-adjusted profiles, contributing to the optimization of microencapsulated diets for bivalve rearing and a better final product.

Freshwater benthic environments are among the major sinks of microplastics (MPs, < 5 mm) sourced on inland anthropogenic activities. The ecotoxicological effects of MPs on benthic macroinvertebrates have been assessed preferably in collectors, shredders, and filter-feeders, but resulting in insufficient knowledge on the potential trophic transfer and its effects on macroinvertebrates with predator behaviour such as planarians. This work evaluated the behavioural (feeding, locomotion), physiological (regeneration) and biochemical responses (aerobic metabolism, energy reserves, oxidative damage) of the planarian Girardia tigrina after consuming contaminated live prey Chironomus riparius larvae previously exposed to microplastics of polyurethane (PU-MPs; 7-9 μm in size; 375 mg PU-MPs/kg). After the feeding period (3 h), planarians consumed 20 % more contaminated prey than uncontaminated prey, probably related to increased curling/uncurling movements of larvae (that might be more appellative to planarians). Histological analysis revealed planarians' limited intake of PU-MPs, mainly detected near the pharynx. The consumption of contaminated prey (and intake of PU-MPs) did not result in oxidative damage but slightly increased the aerobic metabolism and energy reserves which show that the consumption of more prey was sufficient to cope with the potential adverse effects of internalized MPs. Moreover, no effects were observed in the locomotion of planarians in good agreement with the hypothesis of sufficient energy acquired by the exposed planarians. Despite the previous, it seems that the energy acquired was not allocated for planarians' regeneration since a significant delay in the regeneration of the auricles was observed for planarians feeding on contaminated prey. Therefore, further studies should be performed considering the potential long-term effects (i.e., reproduction/fitness) and the effects of MPs that might result from continuous feeding on contaminated prey, representing a more realistic exposure scenario., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Freshwater provides valuable services and functions to humankind. However, macroinvertebrates that underpin the delivery of many of those ecosystem services and functions are under an additional threat caused by microplastic pollution. Chironomids are one of the most abundant groups of macroinvertebrates in these environments and the most sensitive to microplastics. This investigation addressed the effects of polyurethane (PU-MPs; 7.0-9.0 µm) on the chironomid Chironomus riparius at the organism and sub-organism levels. For this purpose, two assays were carried out: (i) addressing the effects of PU-MPs on C. riparius partial life cycle traits (larval size and emergence parameters) in a 28 d assay considering concentrations up to 750 mg/Kg, and (ii) larvae behaviour (locomotion) as well as the biochemical responses (oxidative damage, aerobic energy production, and energy reserves) in a 10 d assay considering an environmentally relevant concentration with no observed effects on C. riparius previous life history traits (no observed effect concentration; NOEC = (375 mg/kg). Exposure to PU-MPs did not affect C. riparius larval length nor cumulative and time to emergence. Conversely, when exposed to an environmentally relevant concentration for 10 days, contaminated larvae were revealed to be lighter (but not smaller nor less nutritionally affected in terms of energy reserves) and more active when foraging, which was reflected in the activation of their aerobic metabolism when assessing the electron transport chain as a proxy. Notwithstanding, PU-MPs did not originate observable energy costs, either on protein, lipid, or sugar contents on contaminated larvae, which may justify the absence of effects on larval growth and emergence. Therefore, the increased production of energy used for the locomotion and functioning of larvae was at the expense of the fraction of energy that should have been allocated for the weight of the individuals. A long-term exposure involving a multigenerational assessment would bring intel on the potential (cumulative) sub-lethal effects of PU-MPs on C. riparius fitness.

Patients with COVID-19 may develop symptoms that interfere with food intake. Systemic inflammatory response associated with physical inactivity and/or immobilization during hospital stay can induce weight and muscle loss leading to sarcopenia and worsening the clinical condition of these patients. The present study identifies the frequency and factors associated with sarcopenia prediction in adult and elderly patients hospitalized for COVID-19. It is a cohort-nested cross-sectional study on adult and elderly patients admitted to wards and intensive care units (ICUs) of 8 hospitals in a northeastern Brazilian state. The study was conducted from June 2020 to June 2021. Sociodemographic, economic, lifestyle, and current and past clinical history variables were collected. Sarcopenia prediction was determined by the Strength, Assistance in walking, Rise from a chair, Climb stairs, and Falls (SARC-F) questionnaire compiled in the Remote-Malnutrition APP (R-MAPP). Patients were diagnosed with sarcopenia when the final score ≥ 4 points. The study included 214 patients with a mean age of 61.76 ± 16.91 years, of which 52.3 % were female and 57.5 % elderly. Sarcopenia prevailed in 40.7 % of the sample. Univariate analysis showed greater probability of sarcopenia in elderly individuals, nonpractitioners of physical activities, hypertensive patients, diabetic patients, and those hospitalized in the ICU. In the multivariate model, the type of hospital admission remained associated with sarcopenia prediction, where patients admitted to the ICU were 1.43 (95 % CI: 1.04; 1.97) more likely to have sarcopenia than those undergoing clinical treatment. Sarcopenia prediction was not associated with patient outcome (discharge, transfer, or death) (p = 0.332). The study highlighted an important percentage of sarcopenia prediction in patients with COVID-19, especially those admitted to the ICU. Additional investigations should be carried out to better understand and develop early diagnostic strategies to assist in the management of sarcopenic patients with COVID-19., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Mercury (Hg) is a non-essential metal that can have toxic effects on the fitness of organisms and tends to bioaccumulate with age and to biomagnify in higher trophic levels. Few studies have assessed oxidative stress and neurotoxicity in deep-water sharks. This study evaluated early ontogenetic changes and physiological effects (antioxidant defences, oxidative damage, aerobic metabolism and neurotransmission functions) of Hg accumulation in the white muscle and brain tissues of the velvet belly lantern shark Etmopterus spinax from the southern Iberian coast (NE Atlantic). Results suggested that the low mercury concentrations observed may induce acute effects in E. spinax before they reach sexual maturity. We found different Hg concentrations in E. spinax: [Hg] males > [Hg] females; [Hg] muscle > [Hg] brain. Females appeared to have higher redox capability translated into higher activities and levels of antioxidant defences than males. However, higher levels of oxidative damage were also observed in females. Whilst the mechanisms underlying these effects remain unknown, these results suggest differences in mercury accumulation between tissues and sex, and potentially deleterious effects on oxidative stress status and neurophysiology of E. spinax, potentially impairing swimming performance and reproduction, which could subsequently impact on the health of both individuals and population., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The widespread use of pesticides in the last decades and their accumulation into the environment gave rise to major environmental and human health concerns. To address this topic, the scientific community pointed out the need to develop methodologies to detect and measure the presence of pesticides in different matrices. Biosensors have been recently explored as fast, easy, and sensitive methods for direct organophosphate pesticides monitoring. Thus, the present work aimed at designing and testing a 3D printed adapter useful on different equipment, and a membrane support to immobilize the esterase-2 from Alicyclobacillus acidocaldarius (EST2) bioreceptor. The latter is labelled with the IAEDANS, a bright fluorescent probe. EST2 was selected since it shows a high specificity toward paraoxon. Our results showed good stability and replicability, with an increasing linear fluorescent intensity recorded from 15 to 150 pmol of labelled EST2. Linearity of data was also observed when using the immobilized labelled EST2 to detect increasing amounts of paraoxon, with a limit of detection (LOD) of 0.09 pmol. This LOD value reveals the high sensitivity of our membrane support when mounted on the 3D adapter, comparable to modern methods using robotic workstations. Notably, the use of an independent support significantly simplified the manipulation of the membrane during experimental procedures and enabled it to match the specificities of different systems. In sum, this work emphasizes the advantages of using 3D printed accessories adapted to respond to the newest research needs., (© 2022. The Author(s).)

Plastic pollution and invasive species are recognised as pervasive threats to marine biodiversity. However, despite the extensive on-going research on microplastics' effects in the biota, knowledge on their combination with additional stressors is still limited. This study investigates the effects of polyamide microplastics (PA-MPs, 1 mg/L), alone and in combination with the toxic exudate from the invasive red seaweed Asparagopsis armata (2%), after a 96 h exposure, in the mussel Mytilus galloprovincialis . Biochemical responses associated with oxidative stress and damage, neurotoxicity, and energy metabolism were evaluated in different tissues (gills, digestive gland, and muscle). Byssus production and PA-MP accumulation were also assessed. Results demonstrated that PA-MPs accumulated the most in the digestive gland of mussels under PA-MP and exudate co-exposure. Furthermore, the combination of stressors also resulted in oxidative damage at the protein level in the gills as well as in a significant reduction in byssus production. Metabolic capacity increased in both PA-MP treatments, consequently affecting the energy balance in mussels under combined stress. Overall, results show a potential increase of PA-MPs toxicity in the presence of A. armata exudate, highlighting the importance of assessing the impact of microplastics in realistic scenarios, specifically in combination with co-occurring stressors, such as invasive species.

The high levels of microplastics (MPs) found in freshwaters, particularly in riverine sediments, may impose a threat to the macroinvertebrate communities with possible consequences at ecosystem-level. The present study aimed to assess the effects of a mixture of different sizes of polyethylene microplastics (PE-MPs) on the composition and structure of macroinvertebrate communities and key-functions, such as primary production and leaf litter decomposition. MPs were mixed in the sediment at three different concentrations (0.1, 1, and 10 g kg -1 ) already found in freshwater sediments to enhance the relevance of the work. After eight days of exposure to PE-MPs, the observed changes in macroinvertebrate community structure were mostly due to the reduction in the abundance of deposit-feeders and grazers that were reduced by ca 31-50% and 34-39%, in the two highest MPs concentrations respectively, in comparison with the control treatment after 8 days of exposure. MPs internal concentrations were detected only in organisms exposed to plastic particles within artificial streams with chironomids and mayflies presenting higher MPs internal levels (average of 115 particles/individual found in chironomids, 166/individual for Baetis sp. and 415 particles/individual for Ephemerella sp.) suggesting higher ingestion of plastic microparticles. Nevertheless, the alterations in the community structure did not translate into impairments in the functional endpoints analysed, leaf litter decomposition and primary production, that were expected due to possible sub-lethal effects (e.g., feeding inhibition) on detritivores and grazers. This study represents one of the few assessments of MPs effects on freshwater benthic macroinvertebrate community structure and the first that simultaneously considered ecosystem-level functional endpoints. Further research combining different microplastics and longer exposure periods are needed to raise knowledge on potential ecological consequences of MPs to freshwaters., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Microplastics and nanomaterials are applied in a myriad of commercial and industrial applications. When leaked to natural environments, such small particles might threaten living organisms' health, particularly when considering their potential combination that remains poorly investigated. This study investigated the physiological and biochemical effects of polyethylene (PE; 64-125 μm in size, 0.1, 1.0, and 10.0 mg·L -1 ) single and combined with an engineered nanomaterial applied in antifouling coatings, the copper-aluminium layered double hydroxides (Cu-Al LDH; 0.33, 1.0, and 3.33 mg·L -1 ) in the flatfish Solea senegalensis larvae (8 dph) after 3 h exposure, in a full factorial design. Particles ingestion, histopathology, and biochemical biomarkers were assessed. Fish larvae presented <1 PE particles in their gut, independently of their concentration in the medium. The histological health index showed minimal pathological alterations at PE combined exposure, with a higher value observed at 1 mg LDH·L -1  × 0.1 mg PE·L -1 . Gut deformity and increased antioxidant defences (catalase), neurotransmission (acetylcholinesterase), and aerobic energy production (electron transport system) were observed at PE ≥ 1.0 mg·L -1 . No oxidative damage (lipid peroxidation) or alterations in the detoxification capacity (glutathione-S-transferase) was observed on single and combined exposures. PE, combined or not with Cu-Al LDH, does not seem to compromise larvae's homeostasis considering levels reported so far in the marine and aquaculture environments. However, harsh effects are expected with MP contamination rise, as projections suggest., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The development of faster, sensitive and real-time methods for detecting organophosphate (OP) pesticides is of utmost priority in the in situ monitoring of these widespread compounds. Research on enzyme-based biosensors is increasing, and a promising candidate as a bioreceptor is the thermostable enzyme esterase-2 from Alicyclobacillus acidocaldarius (EST2), with a lipase-like Ser-His-Asp catalytic triad with a high affinity for OPs. This study aimed to evaluate the applicability of Förster resonance energy transfer (FRET) as a sensitive and reliable method to quantify OPs at environmentally relevant concentrations. For this purpose, the previously developed IAEDANS-labelled EST2-S35C mutant was used, in which tryptophan and IAEDANS fluorophores are the donor and the acceptor, respectively. Fluorometric measurements showed linearity with increased EST2-S35C concentrations. No significant interference was observed in the FRET measurements due to changes in the pH of the medium or the addition of other organic components (glucose, ascorbic acid or yeast extract). Fluorescence quenching due to the presence of paraoxon was observed at concentrations as low as 2 nM, which are considered harmful for the ecosystem. These results pave the way for further experiments encompassing more complex matrices.

Freshwater planarians can be useful for the evaluation of contaminant stress on behavioral endpoints. In this work, we studied the sensitivity of the freshwater planarian Girardia tigrina in response to two model stressors (Hg and NaCl) by evaluating mortality, feeding rate and locomotion. A simple feeding assay with G. tigrina was devised, and an automated tracking system was used to evaluate locomotion. The estimated 96 h LC 50 s were 176.8 μg L -1 of Hg and 6.79 g L -1 of NaCl. Acute effects of Hg also included the disintegration of tissues, and loss of pigmentation. Acute effects of NaCl included motionlessness and rupture of the tegument. Hg and NaCl sub-lethal exposures caused feeding inhibition and locomotion impairment. This study demonstrates the usefulness of planarians for ecotoxicological research and that sensitive behavioral endpoints can evaluate the sub-lethal impacts of stressors to freshwater invertebrates., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The structure of crystalline and amorphous materials in the sodium (Na) super-ionic conductor system Na 1+x Al x Ge 2-x (PO 4 ) 3 with x = 0, 0.4, and 0.8 was investigated by combining (i) neutron and x-ray powder diffraction and pair-distribution function analysis with (ii) 27 Al and 31 P magic angle spinning (MAS) and 31 P/ 23 Na double-resonance nuclear magnetic resonance (NMR) spectroscopy. A Rietveld analysis of the powder diffraction patterns shows that the x = 0 and x = 0.4 compositions crystallize into space group-type R3̄, whereas the x = 0.8 composition crystallizes into space group-type R3̄c. For the as-prepared glass, the pair-distribution functions and 27 Al MAS NMR spectra show the formation of sub-octahedral Ge and Al centered units, which leads to the creation of non-bridging oxygen (NBO) atoms. The influence of these atoms on the ion mobility is discussed. When the as-prepared glass is relaxed by thermal annealing, there is an increase in the Ge and Al coordination numbers that leads to a decrease in the fraction of NBO atoms. A model is proposed for the x = 0 glass in which super-structural units containing octahedral Ge (6) and tetrahedral P (3) motifs are embedded in a matrix of tetrahedral Ge (4) units, where superscripts denote the number of bridging oxygen atoms. The super-structural units can grow in size by a reaction in which NBO atoms on the P (3) motifs are used to convert Ge (4) to Ge (6) units. The resultant P (4) motifs thereby provide the nucleation sites for crystal growth via a homogeneous nucleation mechanism.

Bioinsecticides based on the bacterium Bacillus thuringiensis subsp. israelensis (Bti) are increasingly being applied directly into aquatic compartments to control nuisance mosquitoes and blackflies and are generally considered environmentally friendly alternatives to synthetic insecticides. Bti-based insecticides are considered highly selective, being Diptera-specific, and supposedly decompose rapidly in the environment. Nevertheless, their safety to non-target species and freshwater ecosystems has been questioned by recent studies, which in fact document possible indirect effects in aquatic food webs such as the decrease of prey availability to predators. This work aimed to evaluate the potential effects of a Bti-based insecticide (VectoBac® 12AS) on a freshwater macroinvertebrate community and on stream ecological functions by using artificial microcosm streams. Artificial microcosm streams were colonized with a macroinvertebrate community plus periphyton collected in a stream together with Alnus glutinosa leaf packs. They were exposed for 7 days to different Bti treatments (0, 12, 120, 1200 μg/L), which are within the recommended concentrations of application in aquatic compartments for blackfly and mosquito control. Besides invertebrate community structure and abundance, effects were evaluated regarding leaf decomposition and primary production as measures of ecosystem functioning. Community structure was significantly altered in all Bti treatments after 7 days of exposure, mostly due to a decline in chironomids, followed by oligochaetes, which both belong to the deposit-feeders' functional group. Direct effects on oligochaetes are surprising and require further research. Also, reductions of leaf decomposition due to Bti-induced sublethal effects on shredders (reduced feeding) or mortality of chironomids (that can also feed on coarse organic matter) observed in our study, represent potential indirect effects of Bti in aquatic ecosystems. Our short-exposure experiment evidenced some negative effects on stream benthic invertebrate communities and on ecosystem functioning that must be considered whenever Bti is used in water bodies for blackfly or mosquito control programs., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Coral reefs are declining, affected by climate change and escalating anthropogenic pressures, such as pollution or habitat alteration. Consequently, ecotoxicological assays with tropical corals have increased, specifically towards the study of emergent or persistent pollutants. However, standardized methodology to test for corals is non-existent, and their response to organic solvents, recurrently required in ecotoxicological appraisals, remains unknown. Therefore, we aimed to establish a threshold for the safe use of the selected solvents in ecotoxicological studies with these organisms. We assessed the oxidative stress response (antioxidant response and oxidative damage), cellular energy allocation and photophysiology of the photosynthetic coral Zoanthus sp. (Anthozoa, Hexacorallia) exposed to six doses of three different organic solvents (ethanol, methanol and dimethyl sulfoxide - DMSO). Our results suggest that the coral is more sensitive to methanol and DMSO than to ethanol. Methanol and DMSO LOEC were 0.01 mL L -1 affecting maximum quantum yield (F v /F m ) and glutathione S-transferase (GST) activity, respectively, while for ethanol was 0.03 mL L -1 , influencing F v /F m . Despite the higher tolerance of Zoanthus sp. to ethanol, 2.9 mL L -1 of this organic solvent was the only treatment causing mortality. Based on these findings, thresholds for the use of organic solvents with tropical corals can now be adopted. Nevertheless, species specificities should not be overlooked., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Ocean warming and biological invasions are among the most pervasive factors threatening coastal ecosystems with a potential to interact. Ongoing temperature rise may affect physiological and cellular mechanisms in marine organisms. Moreover, non-indigenous species spread has been a major challenge to biodiversity and ecosystem functions and services. The invasive red seaweed Asparagopsis armata has become successfully established in Europe. Its exudate has been considered deleterious to surrounding native species, but no information exists on its effect under forecasted temperature increase. This study evaluated the combined effects of temperature rise and A. armata exudate exposure on the native mussel Mytilus galloprovincialis . Oxidative stress, neurophysiological and metabolism related biomarkers were evaluated after a 96 h-exposure to exudate (0% and 2%) under present (20 °C) and warming (24 °C) temperature scenarios. Short-term exposure to A. armata exudate affected the oxidative stress status and neurophysiology of the mussels, with a tendency to an increasing toxic action under warming. Significant oxidative damage at protein level was observed in the digestive gland and muscle of individuals exposed simultaneously to the exudate and temperature rise. Thus, under a climate change scenario, it may be expected that prolonged exposure to the combined action of both stressors may compromise M. galloprovincialis fitness and survival.

Virtual reality-based exercise (exergames) improves cognition of the elderly but the neurophysiological effects are poorly understood. The hypothesis herein established is that an ultrafast neurophysiological adaptation occurs in prefrontal cortex of elderly after completion of a single exergames session. To reinforce the aforementioned hypothesis, individuals living in a Long-Term Care Home (LTCH) participated in the study and were randomly allocated into two groups (Virtual Reality Group, VRG, n = 5; and Active Control Group, ACG n = 5). VRG performed six exercises with exergames and ACG performed exercises with the same VRG movements but with no virtual reality. Assessment of frontal cortical activity at rest and during cognitive testing via electroencephalographic activity (EEG) was performed before and immediately after the intervention. Significant decrease in relative power of EEG (RP EEG ) Beta brainwave (-29 ± 18%) in the left prefrontal cortex of VRG compared to ACG (4 ± 9%) (p = 0.007). A slight improvement on semantic fluency in VRG (ES=0.21) was noted. An ultrafast prefrontal cortical adaptation may occur as an effect of a single exergames session, causing a small improvement on cognition of institutionalized elderly., Competing Interests: The authors declare no conflict of interest., (© 2021 The Authors.)

Plant extracts are known for their high content and diversity of polyphenols, which can improve fish oxidative status. A growth trial with Senegalese sole postlarvae (45 days after hatching) fed with one of four experimental diets-control (CTRL), and supplemented with curcumin (CC), green tea (GT), and grape seed (GS) extracts-was performed to assess if supplementation could improve growth performance and oxidative status. At the end of the growth trial, postlarvae were submitted to a thermal stress to assess their robustness. Sole growth was improved by CC and GS diets when compared to those fed the CTRL. CC and CTRL postlarvae presented the lowest oxidative damage (lipid peroxidation and protein carbonylation values). Stress-related biomarkers (heat shock protein 70 and glutathione-S-transferase) decreased in CC fish compared to those fed the CTRL diet, which might be due to a direct antioxidant capacity. In contrast, oxidative damage increased in GT and GS sole reared in standard conditions. However, after a thermal stress, GT and GS diets prevented the increase of protein carbonylation content and the decrease of antioxidant glutathione, depending on exposure time. Overall, dietary supplementation with natural extracts modulated oxidative status and stress response after a short/long-term exposure to temperature.

Mercury (Hg) is a widespread pollutant across estuarine and coastal areas, raising concern on its potential impact on aquatic organisms. Hg may origin from natural and anthropogenic sources, being persistent and potentially toxic to biota, ultimately representing a serious risk to human health. Hg accumulation and toxicity may also induce reactive oxygen species (ROS) production in marine organisms, responsible for cell and tissue damage. Additionally, the temperature is undoubtedly an important environmental factor to consider regarding accumulation, due to its marked influence on the physiology and ecology of aquatic organisms. This study aimed to investigate the effect of different temperature scenarios (15, 20 and 25 °C) on the Hg accumulation in Pomatoschistus microps (Krøyer, 1838) liver and muscle, as well as on oxidative stress responses and energy metabolism, after short-term exposure to a naturally contaminated sediment with an environmentally relevant [Hg] (1.2 μg g -1 ). The results showed that Hg accumulation tends to increase along the temperature gradient with higher values of Hg accumulated in liver than in muscle tissue. The action of antioxidant enzymes and stress proteins seems to be effective in combating oxidative stress in the liver. Despite the action of antioxidant defences in the muscle, oxidative damage was observed at the protein level concomitantly with a decrease in aerobic energy production after exposure to Hg at higher temperatures. These findings are ecologically relevant and highlight the importance of further investigation of combined effects of Hg and other stressors, especially in a scenario of a changing climate where events leading to rapid alterations on water parameters are more frequent., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Paraquat (PQ) is still used in several countries worldwide as an herbicide for weed control in agricultural production, ponds, reservoirs and irrigation canals. Thus, PQ is frequently found in surface water systems and is potentially toxic to aquatic organisms, since it can cause mitochondrial dysfunction altering in the redox state of cells. This study aimed to investigate the chronic effects of PQ to Chironomus riparius and Girardia tigrina, and compare their physiological strategies to cope with environmental stress. The mean emergence time was the most sensitive endpoint for Chironomids, with the lowest observed effect concentrations (LOEC) being 0.02 for males and 0.1 mg PQ L -1 for females. Moreover, PQ reduced the body weight of male and female imagoes, with LOECs of 0.5 and 2.5 mg PQ L -1 , respectively. Paraquat also decreased the respiration rate (LOEC = 2.5 mg PQ L -1 ) and total glutathione (tGSH) content (LOEC = 0.5 mg PQ L -1 ). Thus, the aerobic production of energy was not affected and allowed chironomids to cope with oxidative stress induced by PQ, but with consequent physiological costs in terms of development rates and weight of adults. In planarians, PQ decreased the locomotion and feeding activity, and delayed photoreceptor regeneration (LOECs = 2.5 mg PQ L -1 for all endpoints). Despite increased aerobic energy production (LOEC = 0.5 mg PQ L -1 ), planarians were not able to cope with oxidative stress induced by the highest PQ concentrations, since lipid peroxidation levels were significantly increased (LOEC = 2.5 mg PQ L -1 ) concomitantly with a significant decrease of tGSH (LOEC = 2.5 mg PQ L -1 ). These results showed that planarians were unable to cope with oxidative stress induced by PQ with consequent impairments of behavior and regeneration despite an increased aerobic energy production., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Basidiomata of Phallales have a diversified morphology with adhesive gleba that exudes an odor, usually unpleasant that attracts mainly insects, which disperse the basidiospores. The genus Blumenavia belongs to the family Clathraceae and, based on morphological features, only two species are currently recognized: B. rhacodes and B. angolensis. However, the morphological characters adopted in species delimitations within this genus are inconsistent, and molecular data are scarce. The present study aimed to review and identify informative characters that contribute to the delimitation of Blumenavia species. Exsiccates from America and Africa were analyzed morphologically, and molecularly, using ITS, LSU, ATP6, RPB2 and TEF-1α markers for Maximum Parsimony, Bayesian and Maximum likelihood analyses, and also for coalescent based species delimitations (BP&P), as well as for bPTP, PhyloMap, Topo-phylogenetic and Geophylogenetic reconstructions. According to our studies, seven species can be considered in the genus: B. rhacodes and B. angolensis are maintained, B. usambarensis and B. toribiotalpaensis are reassessed, and three new species are proposed, B. baturitensis Melanda, M.P. Martín & Baseia, sp. nov., B. crucis-hellenicae G. Coelho, Sulzbacher, Grebenc & Cortez, sp. nov., and B. heroica Melanda, Baseia & M.P. Martín, sp. nov. Blumenavia rhacodes is typified by selecting a lectotype and an epitype. Macromorphological characters considered informative to segregate and delimit the species through integrative taxonomy include length of the basidiomata, color, width and presence of grooves on each arm as well as the glebifer position and shape. These must be clearly observed while the basidiomata are still fresh. Since most materials are usually analyzed after dehydration and deposit in collections, field techniques and protocols to describe fugacious characters from fresh specimen are demanded, as well as the use of molecular analysis, in order to better assess recognition and delimitation of species in Blumenavia., Competing Interests: The authors have declared that no competing interests exist.

In the last decades, the use of organic ultraviolet-filters (UV-filters) has increased worldwide, and these compounds are now considered emerging contaminants of many freshwater ecosystems. The present study aimed to assess the effects of 3-(4-methylbenzylidene) camphor (4-MBC) on a freshwater invertebrate community and on associated ecological functions. For that, artificial streams were used, and a natural invertebrate benthic community was exposed to sediments contaminated with two concentrations of 4-MBC. Effects were evaluated regarding macroinvertebrate abundance and community structure, as well as leaf decomposition and primary production. Results showed that the macroinvertebrate community parameters and leaf decomposition rates were not affected by 4-MBC exposure. On the other hand, primary production was strongly reduced. This study highlights the importance of higher tier ecotoxicity experiments for the assessment of the effects of low concentrations of organic UV-filters on freshwater invertebrate community structure and ecosystem functioning., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Microplastics (1 μm-5 mm), a ubiquitous and persistent marine pollutant, pose a severe threat to coral reefs when recently associated with physiological distress and increased diseases on corals. Studies conducted so far have only reported effects on scleractinian species. Knowledge about its effects on other corals (e.g. Order Zoantharia) remains uncovered, and responses at biochemical levels remain poorly documented. This study aimed to assess the potential effects induced by the presence of microplastics (1 and 10 mg L -1 low-density polyethylene, LDPE MP, or polyvinyl chloride, PVC MP) in the tropical and subtropical cosmopolitan species Zoanthus sociatus (order Zoantharia. Anthozoa: Hexacorallia), at organism level (survival and behaviour), endosymbionts (photosynthetic efficiency) and the cellular level (oxidative stress, detoxification capacity and energy metabolism). In a short-term exposure (96 h), this species was more sensitive to PVC MP. The presence of this polymer at a concentration of 10 mg L -1 caused a ten-fold higher adhesion to the coral epidermis, increased photosynthetic efficiency, lipid peroxidation, and antioxidant defences; without, however, inducing energetic costs. Although the observed physiological and biochemical effects did not compromise Z. sociatus survival in the short term, it does not rule out potential long-term (cumulative) effects that could endanger this and other physiologically similar species that underlie coral reefs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

In this study, we synthesize glass-ceramics of the new Na 1+ x Ge 2 (SiO 4 ) x (PO 4 ) 3- x NASICON (Na super-ionic conductor) series to evaluate the effect of Si 4+ /P 5+ substitution on the structural, microstructural, and electrical properties of the NaGe 2 (PO 4 ) 3 system. From X-ray diffraction, the presence of the NASICON phase is confirmed in all glass-ceramics. An expansion of the unit cell volume suggesting an increase in the bottleneck of the NASICON structure is also observed. Impedance spectroscopy allowed the separation of grain and grain boundary contributions. We observe that the grain conductivity is higher than the specific grain boundary conductivity in all of the investigated compositions (0 ≤ x ≤ 0.8). The Si 4+ /P 5+ substitution causes an enhancement of about 2 and 3 orders of magnitude in the grain and specific grain boundary conductivities, respectively. This behavior is attributable to the introduction of new charge carriers (Na + ) in the NASICON structure and a decrease in the activation energy. Finally, the lowest activation energy for grain (0.586 eV) is observed in the x = 0.6 sample, which indicates the easiest displacement of ions in the investigated series, suggesting that this composition presents the most suitable bottleneck size for (Na + ) sodium ion conduction.

The present work describes and illustrates in detail the six zoeal stages of Albunea paretii collected from plankton samples on the Amazon continental shelf in Brazil. The morphological characters are compared with those of other known albuneid larvae. As in other albuneids, the zoeae of Albunea paretii have a semi-globose carapace, a long rostral spine and a pair of long posterolateral spines on pereon, as well as three aesthetascs on the antennule of zoea I, a strong antennal spinous process and a convex telson with a stout posterolateral spine. The zoeal morphology of Albunea paretii and other species of the genus differs, especially regarding the unique presence of the posterolateral spine only on the 5th pleonite in zoea I-IV and the absence in later zoeal stages. The differing features of the albuneid subfamilies are in the telson, antenna and maxilla format and number of setae.

Lutzomyia longipalpis is the main vector of Leishmania infantum, the causative agent of visceral leishmaniasis in the Neotropical realm. Its taxonomic status has been widely discussed once it encompasses a complex of species. The knowledge about the genetic structure of insect vector populations helps the elucidation of components and interactions of the disease ecoepidemiology. Thus, the objective of this study was to genotypically analyze populations of the Lu. longipalpis complex from a macrogeographic perspective using Next Generation Sequencing. Polymorphism analysis of three molecular markers was used to access the levels of population genetic structure among nine different populations of sand flies. Illumina Amplicon Sequencing Protocol® was used to identify possible polymorphic sites. The library was sequenced on paired-end Illumina MiSeq platform. Significant macrogeographical population differentiation was observed among Lu. longipalpis populations via PCA and DAPC analyses. Our results revealed that populations of Lu. longipalpis from the nine municipalities were grouped into three clusters. In addition, it was observed that the levels of Lu. longipalpis population structure could be associated with distance isolation. This new sequencing method allowed us to study different molecular markers after a single sequencing run, and to evaluate population and inter-species differences on a macrogeographic scale., Competing Interests: The authors have declared that no competing interests exist.

Novel species of fungi described in this study include those from various countries as follows: Antarctica , Apenidiella antarctica from permafrost, Cladosporium fildesense from an unidentified marine sponge. Argentina , Geastrum wrightii on humus in mixed forest. Australia , Golovinomyces glandulariae on Glandularia aristigera, Neoanungitea eucalyptorum on leaves of Eucalyptus grandis, Teratosphaeria corymbiicola on leaves of Corymbia ficifolia, Xylaria eucalypti on leaves of Eucalyptus radiata. Brazil , Bovista psammophila on soil, Fusarium awaxy on rotten stalks of Zea mays, Geastrum lanuginosum on leaf litter covered soil, Hermetothecium mikaniae-micranthae (incl. Hermetothecium gen. nov.) on Mikania micrantha, Penicillium reconvexovelosoi in soil, Stagonosporopsis vannaccii from pod of Glycine max. British Virgin Isles , Lactifluus guanensis on soil. Canada , Sorocybe oblongispora on resin of Picea rubens. Chile , Colletotrichum roseum on leaves of Lapageria rosea. China , Setophoma caverna from carbonatite in Karst cave. Colombia , Lareunionomyces eucalypticola on leaves of Eucalyptus grandis. Costa Rica , Psathyrella pivae on wood. Cyprus , Clavulina iris on calcareous substrate. France , Chromosera ambigua and Clavulina iris var. occidentalis on soil. French West Indies , Helminthosphaeria hispidissima on dead wood. Guatemala , Talaromyces guatemalensis in soil. Malaysia , Neotracylla pini (incl. Tracyllales ord. nov. and Neotracylla gen. nov.) and Vermiculariopsiella pini on needles of Pinus tecunumanii. New Zealand , Neoconiothyrium viticola on stems of Vitis vinifera, Parafenestella pittospori on Pittosporum tenuifolium, Pilidium novae-zelandiae on Phoenix sp. Pakistan , Russula quercus-floribundae on forest floor. Portugal , Trichoderma aestuarinum from saline water. Russia , Pluteus liliputianus on fallen branch of deciduous tree, Pluteus spurius on decaying deciduous wood or soil. South Africa , Alloconiothyrium encephalarti, Phyllosticta encephalarticola and Neothyrostroma encephalarti (incl. Neothyrostroma gen. nov.) on leaves of Encephalartos sp., Chalara eucalypticola on leaf spots of Eucalyptus grandis × urophylla, Clypeosphaeria oleae on leaves of Olea capensis, Cylindrocladiella postalofficium on leaf litter of Sideroxylon inerme , Cylindromonium eugeniicola (incl. Cylindromonium gen. nov.) on leaf litter of Eugenia capensis , Cyphellophora goniomatis on leaves of Gonioma kamassi , Nothodactylaria nephrolepidis (incl. Nothodactylaria gen. nov. and Nothodactylariaceae fam. nov.) on leaves of Nephrolepis exaltata , Falcocladium eucalypti and Gyrothrix eucalypti on leaves of Eucalyptus sp., Gyrothrix oleae on leaves of Olea capensis subsp. macrocarpa , Harzia metrosideri on leaf litter of Metrosideros sp., Hippo pota myces phragmitis (incl. Hippo pota myces gen. nov.) on leaves of Phragmites australis , Lectera philenopterae on Philenoptera violacea , Leptosillia mayteni on leaves of Maytenus heterophylla , Lithohypha aloicola and Neoplatysporoides aloes on leaves of Aloe sp., Millesimomyces rhoicissi (incl. Millesimomyces gen. nov.) on leaves of Rhoicissus digitata , Neodevriesia strelitziicola on leaf litter of Strelitzia nicolai , Neokirramyces syzygii (incl. Neokirramyces gen. nov.) on leaf spots of Syzygium sp., Nothoramichloridium perseae (incl. Nothoramichloridium gen. nov. and Anungitiomycetaceae fam. nov.) on leaves of Persea americana , Paramycosphaerella watsoniae on leaf spots of Watsonia sp., Penicillium cuddlyae from dog food, Podocarpomyces knysnanus (incl. Podocarpomyces gen. nov.) on leaves of Podocarpus falcatus , Pseudocercospora heteropyxidicola on leaf spots of Heteropyxis natalensis , Pseudopenidiella podocarpi, Scolecobasidium podocarpi and Ceramothyrium podocarpicola on leaves of Podocarpus latifolius , Scolecobasidium blechni on leaves of Blechnum capense , Stomiopeltis syzygii on leaves of Syzygium chordatum , Strelitziomyces knysnanus (incl. Strelitziomyces gen. nov.) on leaves of Strelitzia alba , Talaromyces clemensii from rotting wood in goldmine, Verrucocladosporium visseri on Carpobrotus edulis. Spain , Boletopsis mediterraneensis on soil, Calycina cortegadensisi on a living twig of Castanea sativa , Emmonsiellopsis tuberculata in fluvial sediments, Mollisia cortegadensis on dead attached twig of Quercus robur , Psathyrella ovispora on soil, Pseudobeltrania lauri on leaf litter of Laurus azorica , Terfezia dunensis in soil, Tuber lucentum in soil, Venturia submersa on submerged plant debris. Thailand , Cordyceps jakajanicola on cicada nymph, Cordyceps kuiburiensis on spider, Distoseptispora caricis on leaves of Carex sp., Ophiocordyceps khonkaenensis on cicada nymph. USA , Cytosporella juncicola and Davidiellomyces juncicola on culms of Juncus effusus , Monochaetia massachusettsianum from air sample, Neohelicomyces melaleucae and Periconia neobrittanica on leaves of Melaleuca styphelioides × lanceolata , Pseudocamarosporium eucalypti on leaves of Eucalyptus sp . , Pseudogymnoascus lindneri from sediment in a mine, Pseudogymnoascus turneri from sediment in a railroad tunnel, Pulchroboletus sclerotiorum on soil, Zygosporium pseudomasonii on leaf of Serenoa repens. Vietnam , Boletus candidissimus and Veloporphyrellus vulpinus on soil. Morphological and culture characteristics are supported by DNA barcodes., (© 2019 Naturalis Biodiversity Center & Westerdijk Fungal Biodiversity Institute.)

Insecticides usually present in low concentrations in streams are known to impair behaviour and development of non-target freshwater invertebrates. Moreover, there is growing awareness that the presence of natural stressors, such as predation risk may magnify the negative effects of pesticides. This is because perception of predation risk can by itself lead to changes on behaviour and physiology of prey species. To evaluate the potential combined effects of both stressors on freshwater detritivores we studied the behavioural and developmental responses of Chironomus riparius to chlorantraniliprole (CAP) exposure under predation risk. Also, we tested whether the presence of a shredder species would alter collector responses under stress. Trials were conducted using a simplified trophic chain: Alnus glutinosa leaves as food resource, the shredder Sericostoma vittatum and the collector C. riparius. CAP toxicity was thus tested under two conditions, presence/absence of the dragonfly predator Cordulegaster boltonii. CAP exposure decreased leaf decomposition. Despite the lack of significance for interactive effects, predation risk marginally modified shredder effect on leaf decomposition, decreasing this ecosystem process. Shredders presence increased leaf decomposition, but impaired chironomids performance, suggesting interspecific competition rather than facilitation. C. riparius growth rate was decreased independently by CAP exposure, presence of predator and shredder species. A marginal interaction between CAP and predation risk was observed regarding chironomids development. To better understand the effects of chemical pollution to natural freshwater populations, natural stressors and species interactions must be taken into consideration, since both vertical and horizontal species interactions play their role on response to stress.

Anthropogenic activities increase pesticide contamination and biological invasions in freshwater ecosystems. Understanding their combined effects on community structure and on ecosystem functioning presents challenges for an improved ecological risk assessment. This study focuses on an artificial stream mesocosms experiment testing for direct and indirect effects of insecticide (chlorantraniliprole - CAP) exposure on the structure of a benthic macroinvertebrate freshwater community and on ecosystem functioning (leaf decomposition, primary production). To understand how predator identity and resource quality alter the community responses to chemical stress, the mediating effects of an invasive predator species (crayfish Procambarus clarkii) and detritus quality (tested by using leaves of the invasive Eucalyptus globulus) on insecticide toxicity were also investigated. Low concentrations of CAP reduced the abundance of shredders and grazers, decreasing leaf decomposition and increasing primary production. Replacement of autochthonous predators and leaf litter by invasive species decreased macroinvertebrate survival, reduced leaf decomposition, and enhanced primary production. Structural equation modeling (SEM) highlighted that CAP toxicity to macroinvertebrates was mediated by the presence of crayfish or eucalypt leaf litter which are now common in many Mediterranean freshwaters. In summary, our results demonstrate that the presence of these two invasive species alters the effects of insecticide exposure on benthic freshwater communities. The approach used here also allowed for a mechanistic evaluation of indirect effects of these stressors and of their interaction on ecosystem functional endpoint, emphasizing the value of incorporating biotic stressors in ecotoxicological experiments.

Pyrethroid insecticides have been used for decades and their worldwide market continues to increase, despite their high toxicity to non-target insects. Recent studies reveal that it is essential to investigate the secondary mechanisms of action of type II pyrethroids to understand their cellular effects on invertebrates. The aim of this study was to evaluate the lethality, behaviour and physiological alterations and energetic costs in caddisfly larvae exposed to environmentally relevant concentrations of esfenvalerate (ESF). ESF caused both mortality and feeding inhibition of exposed caddisfly larvae: nominal ESF 96 h LC 50 was 2.29 μg/L; feeding activity was impaired at concentrations equal or above 0.25 μg/L. At the cellular level, glutathione-S-transferase (GST) activity was increased on caddisfly larvae exposed to 0.25 and 0.5 μg/L ESF, which might contribute to prevent oxidative damage since levels of lipid peroxidation (LPO) were not altered. The energy budget of exposed caddisfly larvae was impaired by exposure to 0.25 μg/L ESF since sugar and protein contents decreased, while a decline of energy consumption was observed. The analysis of feeding, energy reserves and consumption data through structural equation modelling (SEM) allowed to quantify the direct and indirect effects of ESF exposure on bioenergetics of caddisfly larvae. SEM analysis showed a strong negative direct influence of ESF onto feeding activity, sugars content and energy consumption, highlighting a significant positive relationship between sugars and protein contents. These results show that energy expenditure is related to oxidative defense mechanisms induced by ESF stress that may lead to deleterious effects on growth and development., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Caddisflies have been included in ecotoxicological studies because of their sensitivity and ecological relevance. The present study aimed to assess the sublethal effects of an anthranilic diamide insecticide, chlorantraniliprole (CAP), to Sericostoma vittatum. Used worldwide, CAP is a persistent compound that has been found in surface waters at concentrations from 0.1 μg/L to 9.7 μg/L. It targets the ryanodine receptors, and the present ecotoxicological assessment focused on biomarkers related to neurotransmission, biotransformation, oxidative stress damage, and endpoints related to energy processing (feeding, energy reserves, and cellular metabolism). Six days of exposure trials revealed that feeding activity was significantly decreased in S. vittatum larvae exposed to 0.9 μg/L CAP. Concomitantly, a reduction in cellular metabolism and a significant decrease in protein content were also observed in caddisfly larvae exposed to CAP, suggesting metabolic depression. The results show that sublethal concentrations of CAP can cause detrimental sublethal effects on S. vittatum total glutathione content at concentrations as low as 0.2 μg/L. Bioenergetics can be used to assess physiological effects of contaminants, and the present results show that exposure to low, environmentally relevant, concentrations of CAP alter energy acquisition and metabolism in nontarget aquatic insects with potential population level effects. Environ Toxicol Chem 2017;36:1584-1591. © 2016 SETAC., (© 2016 SETAC.)

Phlebotomine sand flies are blood-feeding insects of marked medical and veterinary significance. Investigations on the biology of these insects hold great importance for both ecological and epidemiological purposes. The present work describes a new approach for real-time PCR (qPCR) with SYBR Green ® , named WMG-qPCR, to identify phlebotomine blood meals. The novelty of the assay was to design primers based on the Whole Mitochondrial Genome (WMG) of the potential hosts (human, dog, cat, brown rat and chicken) aiming to amplify through qPCR the regions of mitochondrial DNA (mtDNA) which are less conserved among all species. Initially, the best method for mtDNA extraction to be applied in WMG-qPCR was determined. Afterwards, amplification specificities were accessed by cross-reaction assays with mtDNA samples from all animal species, besides phlebotomine DNA. Finally, the selected primers were also tested for their limit of DNA detection through standard curves constructed by serial dilution of blood DNA obtained for each target animal species. The WMG-qPCR was able to detect as low as 10pL of blood, equivalent to 26, 84, 130, and 320fg DNA of cat, human, dog and rat, respectively. The assay was also capable to amplify as low as 5pL of chicken blood (5pg DNA). In conclusion, WMG-qPCR seems to be a promising tool to identify phlebotomine blood meals, with high species-specificity and sensitivity. Furthermore, as no supplementary techniques are required, this new approach presents minimized costs and simplified technical-training requirements for execution., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Natural populations experiencing intense selection and genetic drift may exhibit limited potential to adapt to environmental change. The present study addresses the following aspects of the "genetic erosion" hypothesis in the midge Chironomus riparius: does long-term mercury (Hg) contamination affect the Hg tolerance of midge populations inhabiting such impacted areas? If so, is there any fitness cost under changing environmental conditions? And does genetic impoverishment influence the susceptibility of C. riparius to cope with environmental stressful conditions? For this end, we tested the acute and chronic tolerance to Hg and salinity in four C. riparius populations differing in their levels of genetic diversity (assessed through microsatellite markers) and past-histories of Hg exposure. Results showed that the midge population collected from a heavily Hg-contaminated site had higher Hg tolerance compared to the population collected from a closely-located reference site suggesting directional selection for Hg-tolerant traits in its native environment despite no genetic erosion in the field. No increased susceptibility under changing environmental conditions of salinity stress was observed. Moreover, results also showed that populations with higher genetic diversity performed better in the partial life-cycle assays providing evidence on the key role that genetic diversity plays as mediator of populations' susceptibility to environmental stress. Our findings are discussed in terms of the suitability of C. riparius as a model organism in evolutionary toxicology studies as well as the validity of ecotoxicological assessments using genetically eroded laboratory populations., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Recent advances in video tracking technologies provide the tools for a sensitive and reproducible analysis of invertebrate activity under stressful conditions nurturing the field of behavioural ecotoxicology. This study aimed to evaluate behavioural responses of the freshwater planarian Dugesia subtentaculata exposed to a model compound, chlorantraniliprole (CAP). This compound is an anthranilic diamide insecticide and due to its neurotoxic action can, at low concentrations, impair behaviour of exposed organisms. Behavioural endpoints measured included feeding and locomotor activities. Feeding responses were based on planarian predatory behaviour using Chironomus riparius larvae as prey. Locomotion was measured by the traditional planarian locomotor velocity (pLMV) assay and additionally using an automated video tracking system using a Zebrabox(®) (Viewpoint, France) device. While feeding and pLMV were significantly impaired at 131.7μg/L CAP, the video tracking system showed that total distance covered by planarians was significantly reduced at concentrations as low as 26.2μg/L CAP. Our results show that more advanced automated video recording systems can be used in the development of sensitive bioassays allowing a reliable, time- and cost-effective quantification of behaviour in aquatic invertebrates. Due to their ecological relevance, behavioural responses should not be disregarded in risk assessment strategies and we advocate the suitability of planarians as suitable organisms for behavioural ecotoxicological studies., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Mercury contamination is a common phenomenon in the marine environment and for this reason it is important to develop cost-effective and relevant tools to assess its toxic effects on a number of different species. To evaluate the possible effects of Hg in the sea snail Gibbula umbilicalis, animals were exposed to increasing concentrations of the contaminant in the ionic form for 96 h. After this exposure period, mortality, feeding and flipping behavior, the activity of the biomarkers glutathione S-transferase, superoxide dismutase, catalase, lactate dehydrogenase and cholinesterase, the levels of lipid peroxidation and cellular energy allocation were measured. After 96 h of exposure to the highest Hg concentration (≈LC20), there was a significant inhibition of the cholinesterase activity as well as impairment in the flipping behavior and post-exposure feeding of the snails. Cholinesterase inhibition was correlated with the impairment of behavioral responses also caused by exposure to Hg. These endpoints, including the novel flipping test, revealed sensitivity to Hg and might be used as relevant early warning indicators of prospective effects at higher biological organization levels, making these parameters potential tools for environmental risk assessment. The proposed test species showed sensitivity to Hg and proved to be a suitable and resourceful species to be used in ecotoxicological testing to assess effects of other contaminants in marine ecosystems., (Copyright © 2014 Elsevier Ltd. All rights reserved.)