The intensification of anomalous events of seawater warming and the co-occurrence with local anthropogenic stressors are threatening coastal marine habitats, including seagrasses, which form extensive underwater meadows. Eutrophication highly affects coastal environments, potentially summing up to the widespread effects of global climate changes. In the present study, we investigated for the first time in seagrasses, the transcriptional response of different plant organs (i.e., leaf and shoot apical meristem, SAM) of the Mediterranean seagrass Posidonia oceanica growing in environments with a different history of nutrient enrichment. To this end, a mesocosm experiment exposing plants to single (nutrient enrichment or temperature increase) and multiple stressors (nutrient enrichment plus temperature increase), was performed. Results revealed a differential transcriptome regulation of plants under single and multiple stressors, showing an organ-specific sensitivity depending on plants' origin. While leaf tissues were more responsive to nutrient stress, SAM revealed a higher sensitivity to temperature treatments, especially in plants already impacted in their native environment. The exposure to stress conditions induced the modulation of different biological processes. Plants living in an oligotrophic environment were more responsive to nutrients compared to plants from a eutrophic environment. Evidences that epigenetic mechanisms were involved in the regulation of transcriptional reprogramming were also observed in both plants’ organs. These results represent a further step in the comprehension of seagrass response to abiotic stressors pointing out the importance of local pressures in a global warming scenario.

In fish, the gut microbiome plays a crucial role in homeostasis and health and is affected by several organic and inorganic environmental contaminants. Amphidromous fish are sentinel species, particularly exposed to these stressors. We used whole metagenome sequencing to characterize the gut microbiome of wild European eels (Anguilla anguilla) at a juvenile stage captured from three sites with contrasted pollution levels in term of heavy metals and persistent organic pollutants. The objectives were to identify what parameters could alter the gut microbiome of this catadromous fish and to explore the potential use of microbiota as bioindicators of environment quality. We identified a total of 1079 microbial genera. Overall, gut microbiome was dominated by Proteobacteria, Firmicutes and Actinobacteria. Alpha and beta diversity were different amongst sites and could be explained by a reduced number of environmental and biological factors, specifically the relative abundance of fish preys in eels’ diet, PCB101, γHCH (lindane), transnonachlor and arsenic. Furthermore, we identified a series of indicator taxa with differential abundance between the three sites. Changes in the microbial communities in the gut caused by environmental pollutants were previously undocumented in European eels. Our results indicate that microbiota might represent another route by which pollutants affect the health of these aquatic sentinel organisms.

During the last decades, the coastal areas of Morocco have witnessed an intense socioeconomic development associated with a continuous population growth and urban extension. This has led to an overexploitation of coastal aquifers leading to a degradation of their water quality. In order to obtain large scale overview on the quality status of Morocco's coastal aquifers (MCA) to assist national water managers to make informed decisions, a comprehensive scrutinization of the MCA against common indicators and using unified methods is essential. In this study, databases from thirteen MCA were analyzed, using multivariate statistical approaches and graphical methods in order to investigate the degree of mineralization in each aquifer and to identify the main salinization processes prevailing in groundwater. The results showed that the dominant groundwater types are Na–Cl, Ca–Mg–Cl, Ca–Mg–SO₄, Ca–Mg–HCO₃ and Ca–HCO₃–Cl. The Gibbs diagram and the seawater contribution (0–37%) indicate that the mineralization is mainly due to the seawater intrusion and water-rock interaction. The salinity degree diagram illustrates that almost all groundwater samples are located in the moderate to very saline zone, indicating that MCA are recharged by water from variable sources. The groundwater quality assessment shows a deterioration, particularly by seawater intrusion and significant nitrate pollution. The temporal evolution confirm that the MCA are influenced by seawater namely in the Atlantic part. The Wilcox and USSL diagram indicate that the majority of sampled water are unsuitable for irrigation uses. In addition, and by referring to the WHO and the Moroccan standards for water potability, large number of samples from the groundwaters of the MCA is not fully adequate for drinking purposes. A set of management actions (e,g., artificial recharge) are proposed in order to mitigate the effect of groundwater overexploitation and seawater intrusion to ensure the sustainability of MCA.

Extended-spectrum β-lactamases (ESBL)-producing Enterobacterales have been classified as critical priority pathogens by the World Health Organization (WHO). ESBL are universally distributed and, in 2006, were firstly reported on a wild animal. Understanding the relative contributions of wild animals to ESBL circulation in the environment is urgently needed. In this work, we have conducted a nationwide study in Portugal to investigate the occurrence of bacteria carrying clinically significant antimicrobial resistance genes (ARG), using widely distributed wild ungulates as model species. A total of 151 antimicrobial resistant-Enterobacterales isolates were detected from 181 wild ungulates: 50% (44/88) of isolates from wild boar (Sus scrofa), 40.3% (25/62) from red deer (Cervus elaphus), 41.4% (12/29) from fallow deer (Dama dama) and 100% (2/2) from mouflon (Ovis aries subsp. musimon). Selected isolates showed a diversified resistance profile, with particularly high values corresponding to ampicillin (71.5%) and tetracycline (63.6%). Enterobacterales strains carried blaTEM, tetA, tetB, sul2, sul1 or dfrA1 ARG genes. They also carried blaCTX₋M-type genes, which are prevalent in human infections, namely CTX-M-14, CTX-M-15 and CTX-M-98. Strikingly, this is the first report of CTX-M-98 in wildlife. Almost 40% (n = 59) of Enterobacterales were multi-drug resistant. The diversity of plasmids carried by ESBL isolates was remarkable, including IncF, K and P. This study highlights the potential role of wild ungulates as environmental reservoirs of CTX-M ESBL-producing E. coli and in the spill-over of AMR bacteria and their determinants. Our findings suggest that wild ungulates are useful as strategic sentinel species of AMR in terrestrial environments, especially in response to potential sources of anthropogenic pollution, providing early warning of potential risks to human, animal and environmental health.

The assessment of contaminant exposure in marine organisms often focuses on the most toxic chemical elements from upper trophic level species. Information on mid-trophic level species and particularly on potentially less harmful elements is lacking. Additionally, microplastics have been considered emergent contaminants in aquatic environments which have not been extensively studied in species from mid-trophic levels in food chains. This study aims to contribute to an overall assessment of environmental impacts of such chemicals in a community of small pelagic fish in the North Atlantic. The concentrations of 16 chemical elements, rarely simultaneously quantified (including minerals, trace elements and heavy metals), and the presence of microplastics were analysed in sardines (Sardina pilchardus) and mackerels (Scomber spp. and Trachurus trachurus) sampled along the Portuguese coast. Biochemical stress assessments and stable isotope analyses were also performed. The chemical element concentrations in S. pilchardus, T. trachurus, and Scomber spp. were relatively low and lower than the levels reported for the same species in the North Atlantic and adjacent areas. No clear relationships were found between chemical elements and oxidative damage in fish. However, the concentration of several chemical elements showed differences among species, being related with the species’ habitat use, trophic niches, and specific feeding strategies. The presence of plastic pieces in the stomachs of 29% of the sampled fishes is particularly concerning, as these small pelagic fish from mid-trophic levels compose a significant part of the diet of humans and other top predators. This study highlights the importance of multidisciplinary approaches focusing on the individual, including position data, stable isotopes, and oxidative stress biomarkers as complementary tools in contamination assessment of the marine mid-trophic levels in food chains.

Pollinating bees are stressed by highly variable environmental conditions, malnutrition, parasites and pathogens, but may also by getting in contact with microorganisms or entomopathogenic nematodes that are used to control plant pests and diseases. While foraging for water, food, or nest material social as well as solitary bees have direct contact or even consume the plant protection product with its active substance (e.g., viruses, bacteria, fungi, etc.). Here, we summarize the results of cage, microcolony, observation hive assays, semi-field and field studies using full-size queen-right colonies. By now, some species and subspecies of the Western and Eastern honey bee (Apis mellifera, A. cerana), few species of bumble bees, very few stingless bee species and only a single species of leafcutter bees have been studied as non-target host organisms. Survival and reproduction are the major criteria that have been evaluated. Especially sublethal effects on the bees' physiology, immune response and metabolisms will be targets of future investigations. By studying infectivity and pathogenic mechanisms, individual strains of the microorganism and impact on different bee species are future challenges, especially under field conditions. Overall, it became evident that honey bees, bumble bees and few stingless bee species may not be suitable surrogate species to make general conclusions for biological mechanisms of bee-microorganism interactions of other social bee species. Solitary bees have been studied on leafcutter bees (Megachile rotundata) only, which shows that this huge group of bees (∼20,000 species worldwide) is right at the beginning to get an insight into the interaction of wild pollinators and microbial plant protection organisms.

This study reports the results obtained from toxicological analyses of different types of baits referred to the laboratory of the Toxicology Area (Faculty of Veterinary Medicine, Cáceres, Spain) over a 17-year period (2002–2018). These baits were suspicious materials found in the environment of the region of Extremadura (Western Spain), where such malpractices are a problem to be addressed, as wide livestock farming and hunting activities are combined with a significant wealth of wildlife (especially birds of prey). A total of 246 baits, including 32 commercial chemical products to be used in baits, were analysed. Samples from 183 cases were received and classified according to the material used for their preparation and the toxic substance found. Overall, the most common bait consisted of meat preparations (56.3% of cases) intended to eliminate predators considered ‘annoying’ for livestock and hunting practices, such as carnivores and scavengers. It should be noted that contact baits (as fenthion-impregnated perches) were also detected (7.6%). Regarding the substances detected, anticholinesterase compounds (organophosphates and carbamates) were the most commonly used substances for the preparation of baits (detected in 85.3% of positive baits). Moreover, 8% of the positive baits presented more than one toxic substance in their composition. Due to the types of toxic compounds and the methods used to prepare the baits, this study shows that the malicious use of highly toxic substances in the environment to kill wildlife is a common and current issue and poses a serious risk to different species.

In estuarine ecosystems, bivalves experience large pH fluctuations caused by the anthropogenic elevation of atmospheric CO₂ and Cu pollution. This study investigates whether Cu toxicity increases indiscriminately in two bivalve species from different estuarine habitats as a result of elevated Cu bioaccumulation in acidified seawater. This was carried out by evaluating the effects of Cu exposure on two bivalve species (clams and scallops) for 28 d, at a series of gradient pH levels (pH 8.1, 7.8, and 7.6). The results demonstrated an increase in the Cu content in the soft tissues of clams and scallops in acidified seawater. Cu toxicity increased under acidified seawater by affecting the molecular pathways, physiological function, biochemical responses, and health status of clams and scallops. An iTRAQ-based quantitative proteomic analysis showed increased protein turnover, disturbed cytoskeleton and signal transduction pathways, apoptosis, and suppressed energy metabolism pathways in the clams and scallops under joint exposure to ocean acidification and Cu. The integrated biomarker response results suggested that scallops were more sensitive to Cu toxicity and/or ocean acidification than clams. The proteomic results suggested that the increased energy metabolism and suppressed protein turnover rates may contribute to a higher resistivity to ocean acidification in clams than scallops. Overall, this study provides molecular insights into the distinct sensitivities between two bivalve species from different habitats under exposure to ocean acidification and/or Cu. The findings emphasize the aggravating impact of ocean acidification on Cu toxicity in clams and scallops. The results show that ocean acidification and copper pollution may reduce the long-term viability of clams and scallops, and lead to the degradation of estuarine ecosystems.

More than 20 years after the Water Framework Directive was adopted, there are still major gaps in the sanitary status of small rivers and waterbodies at the head of basins. These small streams supply water to a large number of wetlands that support a rich biodiversity. Many of these waterbodies are fishponds whose production is destined for human consumption or for the restocking of other aquatic environments. However, these ecosystems are exposed to contaminants, including pesticides and their transformation products. This work aims to provide information on the distribution, diversity, and concentrations of agricultural contaminants in abiotic and biotic compartments from a fishpond located at the head of watersheds. A total of 20 pesticides and 20 transformation products were analyzed by HPLC-ESI-MS/MS in water and sediment sampled monthly throughout a fish production cycle, and in three fish species at the beginning and end of the cycle.The highest mean concentrations were found for metazachlor-OXA (519.48 ± 56.52 ng.L⁻¹) in water and benzamide (4.23 ± 0.17 ng g⁻¹ dry wt.) in sediment. Up to 20 contaminants were detected per water sample and 26 per sediment sample. The transformation products of atrazine (banned in Europe since 2003 but still widely used in other parts of the world), flufenacet, imidacloprid (banned in France since 2018), metazachlor, and metolachlor were more concentrated than their parent compounds. Fewer contaminants were detected in fish and principally prosulfocarb accumulated in organisms during the cycle.Our work brings innovative data on the contamination of small waterbodies located at the head of a basin. The transformation products with the highest frequency of occurrence and concentrations should be prioritized for further environmental monitoring studies, and specific toxicity thresholds should be defined. Few contaminants were found in fish, but the results challenge the widely use of prosulfocarb.

The number of published studies evaluating the effects of microplastics (MPs) in fish has increased in the last decade. However, of the available studies, few have explored the long-term effects of MPs on fish growth and reproduction and have resorted to MPs in the form of μm-sized beads/microspheres. In this study, 6-10 day-old post-hatch medaka (Oryzias latipes) fish were exposed to 50 (i.e. 1X) and 500 (i.e. 10X) μg of heterogeneously sized and irregularly shaped virgin polystyrene (PS) MP particles (200-μm range)/L for 150 days. These concentrations corresponded to respective daily mean values of 247 and 3087 particles/L administered through the diet. The PS MPs dietary exposure resulted in body burdens of 114 and 440 particles/g fish on day 50, and of 78 and 173 particles/g fish on day 100 since the respective exposures to the 1X and the 10X treatments started. The biometric analyses found no incidence of PS MPs ingestion on overall fish growth and development. The histological survey in the 10X group did not reveal alterations in gills or in the digestive tract. Mild alterations in other organs were seen and included increased fluid material in the peritoneal cavity, glomerular and tubular alterations in kidneys, and differences in the diameter of the thyroid follicles and thickness of the follicular epithelial cells. The initial days of the reproductive phase revealed MP-related differences in the number of gravid females, fecundity, and fertilization rates. Overall, these values reverted to normal rates throughout the succeeding days. No significant effects of PS MPs exposure were evidenced on offspring success. The 150-day PS MPs dietary exposure used in this study provided clues of histological effects and a reproduction delay. However, it did not seem to compromise overall growth/thriving and the ongoing reproduction.