We determined the distribution and temporal variation of Carbapenem Resistant Enterobacterales (CRE), carbapenemase-encoding genes and other antibiotic resistance genes (ARGs) in a highly polluted river (Lis River; Portugal), also assessing the potential influence of water quality to this distribution. Water samples were collected in two sampling campaigns performed one year apart (2018/2019) from fifteen sites and water quality was analyzed. CRE were isolated and characterized. The abundance of four ARGs (blaNDM, blaKPC, tetA, blaCTX₋M), two Microbial Source Tracking (MST) indicators (HF183 and Pig-2-Bac) and the class 1 integrase gene (IntI1) was measured by qPCR. confirmed the poor quality of the Lis River water, particularly in sites near pig farms. A collection of 23 CRE was obtained: Klebsiella (n = 19), Enterobacter (n = 2) and Raoultella (n = 2). PFGE analysis revealed a clonal relationship between isolates obtained in different sampling years and sites. All CRE isolates exhibited multidrug resistance profiles. Klebsiella and Raoultella isolates carried blaKPC while Enterobacter harbored blaNDM. Conjugation experiments were successful for only four Klebsiella isolates. All ARGs were detected by qPCR on both sampling campaigns. An increase in ARGs and IntI1 abundances was detected in sites located downstream of wastewater treatment plants. Strong correlations were observed between blaCTX₋M, IntI1 and the human-pollution marker HF183, and also between tetA and the pig-pollution marker Pig-2-bac, suggesting that both human- and animal-derived pollution in the Lis River are a potential source of ARGs. Plus, water quality parameters related to eutrophication and land use were significantly correlated with ARGs abundances. Our findings demonstrated that the Lis River encloses high levels of antibiotic resistant bacteria and ARGs, including CRE and carbapenemase-encoding genes. Overall, this study provides a better understanding on the impacts of water pollution resulting from human and animal activities on the resistome of natural aquatic systems.

The environment is considered a major reservoir of antimicrobial resistant microorganisms (AMR) and antimicrobial resistance genes (ARG). Colistin, a “last resort” antibiotic, is used for the treatment of severe infections caused by multidrug-resistant Gram-negative bacteria. The global dissemination of mobile colistin resistance genes (mcr) in natural and non-natural environments is a major setback in the fight against antimicrobial resistance. Hitherto, there is a limited number of studies screening this resistance determinant in bacteria from wildlife. In this study, we describe for the first time the detection of plasmid-mediated colistin resistance in Escherichia coli from wild ungulates in Portugal, which are also widely distributed across Europe. This information is critical to identify the importance of ungulates in the dissemination of resistant bacteria, and their corresponding genes, across the environment. Here, 151 resistant-Enterobacteriaceae isolated from 181 samples collected from different wild ungulate species throughout Portugal were screened for mcr genes. Four mcr-1-positive Escherichia coli were detected from four fallow deer individuals that were sampled in the same hunting ground. These four isolates harboured mcr-1-related IncP plasmids belonging to sequencing types ST155, ST533 and ST345 (n = 2), suggesting bacterial and/or plasmid circulation. All mcr-1-positive E. coli also showed other resistance phenotypes, including MDR, including the B1 commensal phylogenetic profile. All mcr-1-positive E. coli show additional resistance phenotypes, including MDR, including the B1 commensal phylogenetic profile. Our findings are upsetting, highlighting the global dissemination of colistin resistance genes in the whole ecosystem, which, under the One Health framework, emphasizes the urgent need for effective implementation of AMR surveillance and control in the human-animal-environment interfaces.

Multidisciplinary approaches are essential to diligently assess environmental health status of ecosystems. In this study, year-round chemical elements' exposure and impacts were assessed on the wide-ranging Cory's shearwater Calonectris borealis breeding in Berlenga Island, offshore Portugal, North Atlantic Ocean. The aim was to identify potential contamination and oxidative stress sources associated with trophic ecology, habitat and spatial use, and foraging patterns. A set of 20 chemical elements were quantified, along with oxidative stress biomarkers, stable isotope analyses, and GPS tracking data. Birds presented higher accumulation to some non-essential elements along the year (i.e. arsenic, As; cadmium, Cd; mercury, Hg; lead, Pb; and strontium, Sr), in which concentrations were similar or surpassed other procellariform seabird populations all over the world. No significant differences were found for any of the elements between different periods within the breeding season, with exception of Hg. However, a Principal Component Analysis taking into consideration a group of elements showed differences between pre-laying and chick-rearing periods, with overall higher concentrations in the former. Individuals spending more time engaging in an intensive search for food, and in more coastal environments, presented overall higher element concentrations, and particularly Hg. Contrary to expectations, no relationships were found between chemical elements and oxidative stress. On the other hand, spatial use and foraging patterns of Cory's shearwaters influenced their oxidative stress responses. Our results highlight the need for multidisciplinary approaches to deepen understanding of the large-scale vulnerability of bioindicators such as seabirds and, by extension, the overall environmental health of ecosystems in which they rely.

Endocrine disrupting chemicals (EDCs) include a wide array of pollutants, such as some metals and other toxic elements, which may cause changes in hormonal homeostasis. In addition to affecting physiology of individuals directly, EDCs may alter the transfer of maternal hormones to offspring, i.e. causing transgenerational endocrine disruption. However, such effects have been rarely studied, especially in wild populations. We studied the associations between environmental elemental pollution (As, Cd, Cu, Ni, Pb) and maternally-derived egg thyroid hormones (THs) as well as nestling THs in great tits (Parus major) using extensive sampling of four pairs of polluted and reference populations across Europe (Finland, Belgium, Hungary, Portugal). Previous studies in these populations showed that breeding success, nestling growth and adult and nestling physiology were altered in polluted zones compared to reference zones. We sampled non-incubated eggs to measure maternally-derived egg THs, measured nestling plasma THs and used nestling faeces for assessing local elemental exposure. We also studied whether the effect of elemental pollution on endocrine traits is dependent on calcium (Ca) availability (faecal Ca as a proxy) as low Ca increases toxicity of some elements. Birds in the polluted zones were exposed to markedly higher levels of toxic elements than in reference zones at the populations in Finland, Belgium and Hungary. In contrast to our predictions, we did not find any associations between overall elemental pollution, or individual element concentrations and egg TH and nestling plasma TH levels. However, we found some indication that the effect of metals (Cd and Cu) on egg THs is dependent on Ca availability. In summary, our results suggest that elemental pollution at the studied populations is unlikely to cause overall TH disruption and affect breeding via altered egg or nestling TH levels with the current elemental pollution loads. Associations with Ca availability should be further studied.

Due to its dependence on fossil fuel combustion, emissions from the marine transport sector can significantly contribute to air pollution. This work aims to evaluate the impact of maritime transport emissions on air quality in Portugal using a numerical air quality modelling approach, with high-resolution emission data. Emissions from the European TNO inventory were compiled and pre-processed at hourly and high spatial (∼3 × 3 km²) resolutions. Scenarios with and without these maritime emissions were then simulated with the WRF-CHIMERE modelling system, extensively tested and validated for Portugal domain, in order to evaluate their impact on air quality. A simulation was performed for one year (2016) and the resulting differences were analysed in terms of spatial distribution, time series and deltas. The main deltas for NO₂ and PM10 are located over international shipping routes and major ports, while O₃ concentrations are impacted in a larger area. The modelling results also indicate that shipping emissions are responsible for deltas in the concentration of NO₂ higher than 20% over specific urban areas located in the west coast of Portugal, and less than 5% for PM10. For O₃ the relative contribution is low (around 2%) but this contribution is also observed at locations more than 50 km from the coast.

In the last few years, ship emissions have attracted growing attention in the scientific community. The main reason is the constant increase of marine emissions over the last twenty years due to the intensification of port traffic. Thus, this study aimed to evaluate ship emissions (PM10, PM2.5, NOx, SO2, CO, CO2, N2O CH4, NMVOC, and HC) through the activity-based methodology in four of the main ports of Portugal (Leixões, Setúbal, Sines and Viana do Castelo) during 2013 and 2014. The analysis was performed according to ship types (bulk carrier, container, general cargo, passenger, Ro-Ro cargo, tanker and others) and operational modes (manoeuvring, hotelling and during cruising). Results indicated that tankers were the largest emitters in two of the four analysed ports. Regarding cruising emissions, container ships were the largest emitters. . CO2, NOx and SO2 estimated emissions represented more than 95% of the cruising and in-port emissions. Results were also compared with the total national emissions reported by the Portuguese Environment Agency, and if the in-port emissions estimated in the present study would have been taken into account to these totals, emissions of NOx and SO2 would increase 15% and 24% in 2013 and 16% and 28% in 2014. Summing up ships seem to be an important source of air pollution, mainly regarding NOx and SO2.

Carbon dioxide is one of the main soil gases released silently and permanently in diffuse degassing areas, both in volcanic and non-volcanic zones. In the volcanic islands of the Azores (Portugal) several villages are located over diffuse degassing areas. Lethal indoor CO2 concentrations (higher than 10 vol %) were measured in a shelter located at Furnas village, inside the caldera of the quiescent Furnas Volcano (S. Miguel Island). Hazardous CO2 concentrations were detected not only underground, but also at the ground floor level. Multivariate regression analysis was applied to the CO2 and environmental time series recorded between April 2008 and March 2010 at Furnas village. The results show that about 30% of the indoor CO2 variation is explained by environmental variables, namely barometric pressure, soil water content and wind speed. The highest indoor CO2 concentrations were recorded during bad weather conditions, characterized by low barometric pressure together with rainfall periods and high wind speed. In addition to the spike-like changes observed on the CO2 time series, long-term oscillations were also identified and appeared to represent seasonal variations. In fact, indoor CO2 concentrations were higher during winter period when compared to the dry summer months. Considering the permanent emission of CO2 in various volcanic regions of the world, CO2 hazard maps are crucial and need to be accounted by the land-use planners and authorities.

This work reports changes in suspended particulate matter, turbidity, dissolved Cr, Ni, Cu, Cd, Hg and Pb concentrations, and phytoplankton biomass and composition during a 5-month period dredging operation, in a trace element contaminated area of the Tagus estuary (Portugal). Phytoplankton biomass, diatom:other groups ratio, benthic:pelagic diatom ratio, Margalef's, Simpson's diversity, Shannon–Wiever's, and Warwick and Clarke's taxonomic diversity and distinctness indices, and individual taxa were investigated as indicators of dredging induced changes. Significant rise in sediment resuspension and trace element mobilisation caused by dredging influenced the community structure but not the overall biomass. Benthic diatom displacement into the water column maintained species diversity, and therefore, none of the indices highlighted community changes. Contrastingly, diatom:other groups ratio and benthic:pelagic diatom ratio were reliable indicators for the assessment of dredging induced changes. A shift in composition towards species less susceptible to trace elements was observed, disclosing some individual taxa as potential indicators.

Pools of Zn, Cu, Cd and Co in leaf, stem and root tissues of Sarcocornia fruticosa, Sarcocornia perennis, Halimione portulacoides and Spartina maritima were analyzed on a bimonthly basis, in a Tagus estuary salt marsh. All the major concentrations were found in the root tissues, being the concentrations in the aboveground organs neglectable for sediment budget proposes, as seen by the low root-aboveground translocation. Metal annual accumulation, root turnovers and cycling coefficients were also assessed. S. maritima showed the higher root turnovers and cycling coefficients for most of the analyzed metals, making this a phytostabilizer specie. By contrast the low root turnover, cycling coefficient and low root necromass generation makes S. perennis the most suitable specie for phytoremediation processes. Although the high amounts of metal return to the sediments, due to root senescence, salt marshes can still be considered sinks of heavy metals, cycling heavy metals mostly between sediment and root.

The study presented here searched for the level of taxonomic resolution required to detect the effects of low-level chronic pollution on estuarine meiobenthic communities. Meiofauna from two sites, with special attention to harpacticoid copepods, was analysed at different taxonomic levels of aggregation using uni- and multivariate methods. Adaptation processes that could buffer biodiversity disruptions were also considered through the analysis of fitness-related and tolerance traits in the harpacticoid copepod Paronychocamptus nanus. Results showed that uni- and multivariate analyses could be inadequate when assessing subtle anthropogenic contamination. Instead, the assessment of inter-population differences in tolerance to the main source of stress rises as a required procedure if potential effects of this type of contamination are being investigated. Specifically, a 96 h acute toxicity test performed with populations from the affected site appears as a faster and reliable general tool to assess impacts of low-level chronic pollution in estuaries.