Incorporation of sediment- and soil-specific aspects in the Criteria for Reporting and Evaluating Ecotoxicity Data (CRED).

In environmental risk assessment either for registration purposes or for retrospective assessments of monitoring data, the hazard assessment is predominantly based on effect data from ecotoxicity studies. Most regulatory frameworks require studies used for risk assessment to be evaluated for reliability and relevance. Historically, the Klimisch methodology was used in many regulatory procedures where reliability needed to be evaluated. More recently, the Criteria for Reporting and Evaluating Ecotoxicity Data (CRED) have been developed for aquatic ecotoxicity studies, providing more detailed guidance on the evaluation and reporting of not only the reliability but also the relevance of a scientific study. Here, we discuss the application of the CRED methodology for assessing sediment and soil ecotoxicity studies, addressing important sediment- and soil-specific criteria that should be included as part of the CRED evaluation system. We also provide detailed recommendations for the design and reporting of sediment and soil toxicity studies that can be used by scientists and researchers wishing to contribute ecotoxicological data for effect assessments carried out within regulatory frameworks. Integr Environ Assess Manag 2024;00:1-13. © 2024 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Identification of the Most Immunoreactive Antigens of Candida auris to IgGs from Systemic Infections in Mice.

The delay in making a correct diagnosis of Candida auris causes concern in the healthcare system setting, and immunoproteomics studies are important to identify immunoreactive proteins for new diagnostic strategies. In this study, immunocompetent murine systemic infections caused by non-aggregative and aggregative phenotypes of C. auris and by Candida albicans and Candida haemulonii were carried out, and the obtained sera were used to study their immunoreactivity against C. auris proteins. The results showed higher virulence, in terms of infection signs, weight loss, and histopathological damage, of the non-aggregative isolate. Moreover, C. auris was less virulent than C. albicans but more than C. haemulonii. Regarding the immunoproteomics study, 13 spots recognized by sera from mice infected with both C. auris phenotypes and analyzed by mass spectrometry corresponded to enolase, phosphoglycerate kinase, glyceraldehyde-3-phosphate dehydrogenase, and phosphoglycerate mutase. These four proteins were also recognized by sera obtained from human patients with disseminated C. auris infection but not by sera obtained from mice infected with C. albicans or Aspergillus fumigatus. Spot identification data are available via ProteomeXchange with the identifier PXD049077. In conclusion, this study showed that the identified proteins could be potential candidates to be studied as new diagnostic or even therapeutic targets for C. auris.

Toxicogenomics of the Freshwater Oligochaete, Tubifex tubifex (Annelida, Clitellata), in Acute Water-Only Exposure to Arsenic.

A toxicogenomic approach was used for toxicity evaluation of arsenic in the aquatic environment, and differential gene expression was investigated from 24 h and 96 h water-only acute toxicity tests with the aquatic oligochaete, Tubifex tubifex (Annelida, Clitellata). Several toxicological endpoints (survival and autotomy) of the oligochaete and tissue residues were measured, and dose-response modelling of gene expression data was studied. A reference transcriptome of the aquatic oligochaete, T. tubifex, was reconstructed for the first time, and genes related to cell stress response (Hsc70, Hsp10, Hsp60, and Hsp83), energy metabolism (COX1), oxidative stress (Cat, GSR, and MnSOD), and the genes involved in the homeostasis of organisms (CaM, RpS13, and UBE2) were identified and characterised. The potential use of the genes identified for risk assessment in freshwater ecosystems as early biomarkers of arsenic toxicity is discussed.

Candida albicans increases the aerobic glycolysis and activates MAPK-dependent inflammatory response of liver sinusoidal endothelial cells.

The liver, and more specifically, the liver sinusoidal endothelial cells, constitute the beginning of one of the most important responses for the elimination of hematogenously disseminated Candida albicans. Therefore, we aimed to study the mechanisms involved in the interaction between these cells and C. albicans. Transcriptomics-based analysis showed an increase in the expression of genes related to the immune response (including receptors, cytokines, and adhesion molecules), as well as to aerobic glycolysis. Further in vitro analyses showed that IL-6 production in response to C. albicans is controlled by MyD88- and SYK-pathways, suggesting an involvement of Toll-like and C-type lectin receptors and the subsequent activation of the MAP-kinases and c-Fos/AP-1 transcription factor. In addition, liver sinusoidal endothelial cells undergo metabolic reprogramming towards aerobic glycolysis induced by C. albicans, as confirmed by the increased Extracellular Acidification Rate and the overexpression of enolase (Eno2), hexonikase (Hk2) and glucose transporter 1 (Slc2a1). In conclusion, these results indicate that the hepatic endothelium responds to C. albicans by increasing aerobic glycolysis and promoting an inflammatory environment.

Microbiota and fungal-bacterial interactions in the cystic fibrosis lung.

The most common genetic hereditary disease affecting Caucasians is cystic fibrosis (CF), which is caused by autosomal recessive mutations in the CFTR gene. The most serious consequence is the production of a thick and sticky mucus in the respiratory tract, which entraps airborne microorganisms and facilitates colonization, inflammation and infection. Therefore, the present article compiles the information about the microbiota and, particularly, the inter-kingdom fungal-bacterial interactions in the CF lung, the molecules involved and the potential effects that these interactions may have on the course of the disease. Among the bacterial compounds, quorum sensing-regulated molecules such as homoserine lactones, phenazines, rhamnolipids, quinolones and siderophores (pyoverdine and pyochelin) stand out, but volatile organic compounds, maltophilin and CF-related bacteriophages are also explained. These molecules exhibit diverse antifungal mechanisms, including iron starvation and induction of reactive oxygen and nitrogen species production. The fungal compounds are less studied, but they include cell wall components, siderophores, patulin and farnesol. Despite the apparent competition between microorganisms, the persistence of significant rates of bacterial-fungal co-colonization in CF suggests that numerous variables influence it. In conclusion, it is crucial to increase scientific and economic efforts to intensify studies on the bacterial-fungal inter-kingdom interactions in the CF lung.

Dot Immunobinding Assay for the Rapid Serodetection of Scedosporium/Lomentospora in Cystic Fibrosis Patients.

The detection of Scedosporium/Lomentospora is still based on non-standardized low-sensitivity culture procedures. This fact is particularly worrying in patients with cystic fibrosis (CF), where these fungi are the second most common filamentous fungi isolated, because a poor and delayed diagnosis can worsen the prognosis of the disease. To contribute to the discovery of new diagnostic strategies, a rapid serological dot immunobinding assay (DIA) that allows the detection of serum IgG against Scedosporium/Lomentospora in less than 15 min was developed. A crude protein extract from the conidia and hyphae of Scedosporium boydii was employed as a fungal antigen. The DIA was evaluated using 303 CF serum samples (162 patients) grouped according to the detection of Scedosporium/Lomentospora in the respiratory sample by culture, obtaining a sensitivity and specificity of 90.48% and 79.30%, respectively; positive and negative predictive values of 54.81% and 96.77%, and an efficiency of 81.72%. The clinical factors associated with the results were also studied using a univariate and a multivariate analysis, which showed that Scedosporium/Lomentospora positive sputum, elevated anti-Aspergillus serum IgG and chronic Pseudomonas aeruginosa infection were significantly associated with a positive result in DIA, while Staphylococcus aureus positive sputum showed a negative association. In conclusion, the test developed can offer a complementary, rapid, simple and sensitive method to contribute to the diagnosis of Scedosporium/Lomentospora in patients with CF.

Baby and us: Community-based, Feasibility Trial of a Psychosocial Intervention for New Parents and their Infants.

Infancy is a critical period during which major developmental transformations occur. Early parenting is one of the strongest influences on infants' immediate and longer-term outcomes. The transition to parenting can be demanding and stressful for mothers and fathers. This paper reports results from a feasibility study of the Empowering Parents Empowering Communities Baby and Us programme, an 8-week, universal, peer-led parenting programme for new parents living in socially disadvantaged communities. This study is a quasi-experimental, one arm, no control group study, assessing the feasibility and acceptability of Baby and Us. Programme participants (n = 158) completed standardised self-report measures of parent goal attainment, self-efficacy, knowledge about parenting, mental wellbeing, parental confidence, and programme acceptability. We found that recruiting parents from disadvantaged backgrounds was feasible (96% of programmes recruited sufficient parents to proceed, mean = 6.6 parents per programme); parent goals closely matched the aims of the programme; programme completion was high (74%), and self-report measurement completion rates were in line with other large scale community delivered parenting programmes; parents rated the programme as highly satisfactory; and they reported significant improvements in their mental wellbeing, confidence, parenting skills, self-efficacy, and goal attainment. These results provide important data to conduct a full-scale trial of Baby and Us.

Developing As and Cu Tissue Residue Thresholds to Attain the Good Ecological Status of Rivers in Mining Areas.

The study was performed on residue-effects datasets from polluted and unpolluted sites in the Nalón River basin (northern Spain). The effects were measured in terms of alteration of field macroinvertebrate communities, and measured as ecological status scores, and number of families and abundance of Ephemeroptera, Plecoptera and Trichoptera (EPT). Non-linear regression models of the field-measured tissue residues in 10 taxa related to the ecological status of the macroinvertebrate communities were used to derive effective tissue residues (ERs). These were estimated for the good/moderate boundary defined by the ecological quality ratio (EQRs) score and for the 50% reduction of EQR and EPT metrics. As, Cu, Hg and Se ERs were calculated for several macroinvertebrate taxa with different feeding styles. The ER dataset allowed us to estimate As and Cu hazardous concentrations (HC), using species sensitivity distribution models, and were interpreted as community thresholds. Further studies for Hg and Se are needed to complete the database required for HC estimation. The reliability and differences of the several thresholds were tested in a risk assessment using a tissue-residue approach (TRA) conducted with field organisms from Cauxa Creek, a tributary from the same basin exposed to high levels of metals in the sediments due to gold mining activities. This risk assessment identified that As and Cu tissue residues satisfactorily explained the reduction in the ecological status of the macroinvertebrate assemblages. Our results indicate that TRA can help in setting future environmental quality standards for the protection of aquatic biota.

Magnetic, Mechanically Interlocked Porphyrin-Carbon Nanotubes for Quantum Computation and Spintronics.

Atomic-scale reproducibility and tunability endorse magnetic molecules as candidates for spin qubits and spintronics. A major challenge is to implant those molecular spins into circuit geometries that may allow one, two, or a few spins to be addressed in a controlled way. Here, the formation of mechanically bonded, magnetic porphyrin dimeric rings around carbon nanotubes (mMINTs) is presented. The mechanical bond places the porphyrin magnetic cores in close contact with the carbon nanotube without disturbing their structures. A combination of spectroscopic techniques shows that the magnetic geometry of the dimers is preserved upon formation of the macrocycle and the mMINT. Moreover, the metallic core selection determines the spin location in the mMINT. The suitability of mMINTs as qubits is explored by measuring their quantum coherence times (Tm). Formation of the dimeric ring preserves the Tm found in the monomer, which remains in the µs scale for mMINTs. The carbon nanotube is used as vessel to place the molecules in complex circuits. This strategy can be extended to other families of magnetic molecules. The size and composition of the macrocycle can be tailored to modulate magnetic interactions between the cores and to introduce magnetic asymmetries (heterometallic dimers) for more complex molecule-based qubits.

Bioaccumulation and chronic toxicity of arsenic and zinc in the aquatic oligochaetes Branchiura sowerbyi and Tubifex tubifex (Annelida, Clitellata).

Oligochaetes feed on bulk sediment and penetrate the sediment through the construction of burrows, making them especially vulnerable to sediment metal contamination. However, the few oligochaete species that have been tested to date are almost exclusively temperate test species. Although the warmwater adapted species Branchiura sowerbyi has been indicated as a promising candidate for tropical sediment toxicity testing, few (especially chronic) studies have been conducted so far to confirm this. Therefore, the aim of the present study was to evaluate the bioaccumulation and chronic 28d lethal and sublethal toxicity of arsenic (As) and zinc (Zn) to both the warmwater-adapted B. sowerbyi and the coldwater-adapted oligochaete Tubifex tubifex for comparison. Arsenic was more toxic to both oligochaete species than Zn. Inter- and intra-species variability in toxicity values of the two test species and other benthic invertebrates was within an order of magnitude. However, B. sowerbyi was the most sensitive species to As even for sediment concentration (EC50: 36.6 ± 2.1 µg/g and 147.1 ± 21.7 µg/g, for B. sowerbyi and T. tubifex, respectively) and for tissue concentration (ER50: 9.2 ± 0.9 µg/g and 887.0 ± 35.0 µg/g, for B. sowerbyi and T. tubifex, respectively). Finally, the Tissue Residue-effects Approach (TRA) using Effective Tissue Residues appears to be a promising way forward in advancing in this since it considers internal body concentrations.

Proposal of integrative scores and biomonitor selection for metal bioaccumulation risk assessment in mine-impacted rivers.

Development of sound criteria for metal and metalloid bioaccumulation risk assessment in river basins affected by mining activities is a necessary tool to protect the aquatic communities. The aim of this study is to propose integrative scores for tissue residues that are suitable for surveillance programs and readily interpreted in terms of risk assessment in mining impacted rivers. Tissue residues of 7 trace metals and 2 metalloids were measured in ten macroinvertebrate taxa from the Nalón River basin (Spain), affected by Hg, Cu and Au mining activities. Compared with reference sites, biomonitor taxa from Hg and Au mining districts showed the highest bioaccumulation. However, low or non-significant bioaccumulation was found in sites influenced by historical Cu mining. Multivariate analyses (ANOSIM) performed on individual taxa revealed significant differences in tissue residues between sites classified according to their ecological status. The bioaccumulation risk assessment was based on the average ratio of the actual metal tissue residues in each macroinvertebrate taxon to the corresponding Ecological Threshold tissue concentration (Tissue residue Ratio to Threshold, TRT). The suitability of the biomonitors was evaluated using linear regression models fitted to the relationships between TRT scores and site sediment pollution or ecological status scores. Biomonitor selection also considered differences in invertebrate functional traits, which can influence metal and metalloid bioavailability. Site bioaccumulation risk was assessed on an Integrated Tissue concentration score (INTISS), calculated over a selection of the most relevant chemicals (As, Cu and Hg) and 3 biomonitor taxa (Baetidae, Hydropsychidae, Microdrile oligochaetes) comprising a set of feeding styles. Based on INTISS, it was possible to predict community alteration scores, using linear regression models. A comparison of site bioaccumulation and ecological status assessments based on the departure from reference conditions showed that operational monitoring programs in basins impaired by mining can be optimized by combining both approaches.

Exploring attitudes towards use of technology to support stroke survivors living at home: A quantitative and qualitative content analysis study in Spain.

PURPOSE: The aim of this study was to better understand the attitudes towards the use of technology to support chronic stroke survivors in a home-based setting. METHODS: A quantitative study was used on the data obtained from a face to face survey with the sample group, incorporating quantitative statistical analysis. RESULTS: Participants reported positive attitudes towards using technology for their own independent health management. The purpose of the home-based technology was different: source of information, supporting self-management, pharmacological treatment reminders… and differed according to age, educational level and survivor disability. Installing devices and sharing information remains a challenge. CONCLUSIONS: 100% of stroke survivors living in the community with any type of disability, reported that they would like to use technology as a tool to help improve their health status.

The Host Immune Response to Scedosporium/Lomentospora.

Infections caused by the opportunistic pathogens Scedosporium/Lomentospora are on the rise. This causes problems in the clinic due to the difficulty in diagnosing and treating them. This review collates information published on immune response against these fungi, since an understanding of the mechanisms involved is of great interest in developing more effective strategies against them. Scedosporium/Lomentospora cell wall components, including peptidorhamnomannans (PRMs), α-glucans and glucosylceramides, are important immune response activators following their recognition by TLR2, TLR4 and Dectin-1 and through receptors that are yet unknown. After recognition, cytokine synthesis and antifungal activity of different phagocytes and epithelial cells is species-specific, highlighting the poor response by microglial cells against L. prolificans. Moreover, a great number of Scedosporium/Lomentospora antigens have been identified, most notably catalase, PRM and Hsp70 for their potential medical applicability. Against host immune response, these fungi contain evasion mechanisms, inducing host non-protective response, masking fungal molecular patterns, destructing host defense proteins and decreasing oxidative killing. In conclusion, although many advances have been made, many aspects remain to be elucidated and more research is necessary to shed light on the immune response to Scedosporium/Lomentospora.

ELISA Test for the Serological Detection of Scedosporium/Lomentospora in Cystic Fibrosis Patients.

The detection and diagnosis of the opportunistic fungi Scedosporium spp. and Lomentospora prolificans still relies mainly on low-sensitive culture-based methods. This fact is especially worrying in Cystic Fibrosis (CF) patients in whom these fungal species are frequently isolated and may increase the risk of suffering from an infection or other health problems. Therefore, with the purpose of developing a serologic detection method for Scedosporium/Lomentospora, four different Scedosporium boydii protein extracts (whole cell protein extract, secretome, total cell surface and conidial surface associated proteins) were studied by ELISA to select the most useful for IgG detection in sera from CF patients. The four extracts were able to discriminate the Scedosporium/Lomentospora-infected from Aspergillus-infected and non-infected patients. However, the whole cell protein extract was the one selected, as it was the one with the highest output in terms of protein concentration per ml of fungal culture used, and its discriminatory capacity was the best. The ELISA test developed was then assayed with 212 sera from CF patients and it showed to be able to detect Scedosporium spp. and Lomentospora prolificans with very high sensitivity and specificity, 86%-100% and 93%-99%, respectively, depending on the cut-off value chosen (four values were proposed A450nm= 0.5837, A450nm= 0.6042, A450nm= 0.6404, and A450nm= 0.7099). Thus, although more research is needed to reach a standardized method, this ELISA platform offers a rapid, low-cost and easy solution to detect these elusive fungi through minimally invasive sampling, allowing the monitoring of the humoral response to fungal presence.

Identification of Mucor circinelloides antigens recognized by sera from immunocompromised infected mice.

BACKGROUND: Mucor circinelloides is an opportunistic fungus capable of causing mucormycosis, a highly aggressive infection of quick spreading. Besides, it also has a high mortality rate due to late diagnosis and difficult treatment. AIMS: In this study we have identified the most immunoreactive proteins of the secretome and the total protein extract of M. circinelloides using sera from immunocompromised infected mice. METHODS: The proteins of the secretome and the total extract were analyzed by two-dimensional electrophoresis and the most immunoreactive antigens were detected by Western Blot, facing the sera of immunocompromised infected mice to the proteins obtained in both extracts of M. circinelloides. RESULTS: Seven antigens were detected in the secretome extract, and two in the total extract, all of them corresponding only to three proteins. The enzyme enolase was detected in both extracts, while triosephosphate isomerase was detected in the secretome, and heat shock protein HSS1 in the total extract. CONCLUSIONS: In this work the most immunoreactive antigens of the secretome and the total extract of M. circinelloides were identified. The identified proteins are well known fungal antigens and, therefore, these findings can be useful for future research into alternatives for the diagnosis and treatment of mucormycosis.

Home-Based Rehabilitation System for Stroke Survivors: A Clinical Evaluation.

Recently, a home-based rehabilitation system for stroke survivors (Baptista et al. Comput. Meth. Prog. Biomed. 176:111-120 2019), composed of two linked applications (one for the therapist and another one for the patient), has been introduced. The proposed system has been previously tested on healthy subjects. However, for a fair evaluation, it is necessary to carry out a clinical study considering stroke survivors. This work aims at evaluating the home-based rehabilitation system on 10 chronic post-stroke spastic patients. For this purpose, each patient carries out two exercises implying the motion of the spastic upper limb using the home-based rehabilitation system. The impact of the color-based 3D skeletal feedback, guiding the patients during the training, is studied. The Time Variable Replacement (TVR)-based average distance, as well as the average postural angle used in Baptista et al. (Comput. Meth. Prog. Biomed. 176:111-120 2019), are reported to compare the movement and the posture of the patient with and without showing the feedback proposals, respectively. Furthermore, three different questionnaires, specifically designed for this study, are used to evaluate the user experience of the therapist and the patients. Overall, the reported results suggest the relevance of the proposed system for home-based rehabilitation of stroke survivors.

Identification of Mucor circinelloides antigens recognized by sera from immunocompromised infected mice / Identificación de antígenos de Mucor circinelloides reconocidos por sueros de ratones inmunodeprimidos infectados

BACKGROUND: Mucor circinelloides is an opportunistic fungus capable of causing mucormycosis, a highly aggressive infection of quick spreading. Besides, it also has a high mortality rate due to late diagnosis and difficult treatment. AIMS: In this study we have identified the most immunoreactive proteins of the secretome and the total protein extract of M. circinelloides using sera from immunocompromised infected mice. METHODS: The proteins of the secretome and the total extract were analyzed by two-dimensional electrophoresis and the most immunoreactive antigens were detected by Western Blot, facing the sera of immunocompromised infected mice to the proteins obtained in both extracts of M. circinelloides. RESULTS: Seven antigens were detected in the secretome extract, and two in the total extract, all of them corresponding only to three proteins. The enzyme enolase was detected in both extracts, while triosephosphate isomerase was detected in the secretome, and heat shock protein HSS1 in the total extract. CONCLUSIONS: In this work the most immunoreactive antigens of the secretome and the total extract of M. circinelloides were identified. The identified proteins are well known fungal antigens and, therefore, these findings can be useful for future research into alternatives for the diagnosis and treatment of mucormycosis

ANTECEDENTES: Mucor circinelloides es un hongo oportunista causante de la mucormicosis, una infección altamente agresiva y de rápida expansión. Además, también presenta una alta mortalidad debido al diagnóstico tardío y el difícil tratamiento. OBJETIVOS: En este estudio se han identificado las proteínas más inmunorreactivas del secretoma y del extracto total de proteínas de M. circinelloides mediante el uso de sueros obtenidos de ratones inmunodeprimidos infectados. MÉTODOS: Las proteínas del secretoma y del extracto total se analizaron mediante electroforesis bidimensional y se detectaron los antígenos más inmunorreactivos mediante Western Blot, enfrentando el suero de los ratones inmunodeprimidos infectados a las proteínas obtenidas en ambos extractos de M. circinelloides. RESULTADOS: Se identificaron 7 antígenos en el secretoma y 2 en el extracto total, todos ellos correspondientes a 3 proteínas. La enolasa se detectó en ambos extractos, mientras que la triosafosfato isomerasa se detectó en el secretoma, y la proteína de choque térmico HSS1 en el extracto total. CONCLUSIONES: En este trabajo se identificaron los antígenos más inmunorreactivos del secretoma y del extracto total de M. circinelloides. Todas las proteínas identificadas son antígenos fúngicos muy conocidos y, por ello, estos resultados pueden ser de gran utilidad en futuras investigaciones relacionadas con la mejora del diagnóstico y el tratamiento de la mucormicosis

The monoclonal antibody Ca37, developed against Candida albicans alcohol dehydrogenase, inhibits the yeast in vitro and in vivo.

Candida albicans is a commensal yeast able to cause life threatening invasive infections particularly in immunocompromised patients. Despite the availability of antifungal treatments, mortality rates are still unacceptably high and drug resistance is increasing. We, therefore, generated the Ca37 monoclonal antibody against the C. albicans alcohol dehydrogenase (Adh) 1. Our data showed that Ca37 was able to detect C. albicans cells, and it bound to Adh1 in yeast and Adh2 in hyphae among the cell wall-associated proteins. Moreover, Ca37 was able to inhibit candidal growth following 18 h incubation time and reduced the minimal inhibitory concentration of amphotericin B or fluconazole when used in combination with those antifungals. In addition, the antibody prolonged the survival of C. albicans infected-Galleria mellonella larvae, when C. albicans was exposed to antibody prior to inoculating G. mellonella or by direct application as a therapeutic agent on infected larvae. In conclusion, the Ca37 monoclonal antibody proved to be effective against C. albicans, both in vitro and in vivo, and to act together with antifungal drugs, suggesting Adh proteins could be interesting therapeutic targets against this pathogen.

Changes in invertebrate community composition allow for consistent interpretation of biodiversity loss in ecological status assessment.

Biological communities change in response to human alteration. The response of individual taxa and the community can be used to establish preventive criteria to halt further biodiversity deterioration. Here we explore how consistent are the boundaries between Good and Moderate ecological status derived from classification systems used in North-NW Spain: NORThern Spain Indicators system (NORTI), River type specific multimetric (METI) and Iberian Bio-monitoring Working Party (IBMWP), by using common interpretation of normative definitions of Water Framework Directive. We applied the three classifications to a monitoring dataset of Nalón River basin, comprising samples from different stream types and reference conditions. We applied Threshold Indicator Taxa ANalysis to the invertebrate community along the most relevant environmental pressures and biological impairment gradients represented by the Ecological Quality Ratio (EQR) scores of the classification systems. Only NORTI provided a true community ecological threshold and the change point (cp) 95% quantile (Q95%) range of 0.760 was assumed to be the boundary from Good to Moderate (G/M) status, used to standardize the number of taxa loss in all systems. Since the average number of taxa at reference sites was 34, the estimated loss of sensitive taxa was up to 97.1% in IBMWP, 73.5% in METI and 52.9% in NORTI when passing from Good to Moderate status, revealing very permissive boundaries. The loss of common sensitive taxa in NORTI at Q95% was used as G/M threshold and applied to the other classifications, resulting all in a common biodiversity loss of 21% of sensitive taxa richness at values of NORTI-EQR = 0.760, METI-EQR = 0.818 and IBMWP-EQR = 0.753. Results indicate that significant community changes along pressure gradients allow for establishing quantitative criteria consistent with normative definitions. This understanding derived from Directive monitoring programs can assess the risk that invertebrate communities face in terms of species loss derived from anthropogenic pressures.

Exposure to heavy metal-contaminated sediments disrupts gene expression, lipid profile, and life history traits in the midge Chironomus riparius.

Despite the concern about anthropogenic heavy metal accumulation, there remain few multi-level ecotoxicological studies to evaluate their effects in fluvial ecosystems. The toxicity of field-collected sediments exhibiting a gradient of heavy metal contamination (Cd, Pb, and Zn) was assessed in Chironomus riparius. For this purpose, larvae were exposed throughout their entire life cycle to these sediments, and toxic effects were measured at different levels of biological organization, from the molecular (lipidomic analysis and transcriptional profile) to the whole organism response (respiration rate, shape markers, and emergence rate). Alterations in the activity of relevant genes, as well as an increase of storage lipids and decrease in membrane fluidity, were detected in larvae exposed to the most contaminated sediments. Moreover, reduced larval and adult mass, decrease of larval respiration rate, and delayed emergence were observed, along with increased mentum and mandible size in larvae and decreased wing loading in adults. This study points out the deleterious effects of heavy metal exposure at various levels of biological organization and provides some clues regarding the mode of toxic action. This integrative approach provides new insights into the multi-level effects on aquatic insects exposed to heavy metal mixtures in field sediments, providing useful tools for ecological risk assessment in freshwater ecosystems.