Innovative and practical tools for monitoring and assessing biodiversity status and impacts of multiple human pressures in marine systems.

Human activities at sea can produce pressures and cumulative effects on ecosystem components that need to be monitored and assessed in a cost-effective manner. Five Horizon European projects have joined forces to collaboratively increase our knowledge and skills to monitor and assess the ocean in an innovative way, assisting managers and policy-makers in taking decisions to maintain sustainable activities at sea. Here, we present and discuss the status of some methods revised during a summer school, aiming at better management of coasts and seas. We include novel methods to monitor the coastal and ocean waters (e.g. environmental DNA, drones, imaging and artificial intelligence, climate modelling and spatial planning) and innovative tools to assess the status (e.g. cumulative impacts assessment, multiple pressures, Nested Environmental status Assessment Tool (NEAT), ecosystem services assessment or a new unifying approach). As a concluding remark, some of the most important challenges ahead are assessing the pros and cons of novel methods, comparing them with benchmark technologies and integrating these into long-standing time series for data continuity. This requires transition periods and careful planning, which can be covered through an intense collaboration of current and future European projects on marine biodiversity and ecosystem health.

Assessment of the eutrophication status at Mediterranean sub-basin scale, within the European Marine Strategy Framework Directive.

The aim of this work is to define harmonized reference conditions and assessment thresholds for selected criteria elements of the Marine Strategy Framework Directive (MSFD) Descriptor 5 (Eutrophication) in the Eastern Mediterranean Sea and to test if a tool for integrated assessment of the status of marine systems can be used as a common methodological approach. In this frame, we tested two statistical approaches in order to set threshold values for four criteria of Descriptor 5: nutrients, chlorophyll a, transparency and dissolved oxygen in the bottom waters. It is noteworthy that this work revealed the need to apply common procedures in data treatment and assessment evaluation. This is the first attempt to set common methods for the assessment of eutrophication in the Eastern Mediterranean, which is essential in marine environments, especially those shared by several countries. To this end, we have applied common criteria and metrics and established thresholds "Good" and "Moderate" for nutrients, chlorophyll a, transparency and dissolved oxygen in the bottom waters for the different Water Types of the Adriatic and Aegean Seas (I, II, IIIW, IIIE), based on datasets provided by Italy, Slovenia, Croatia and Greece. The selected criteria elements were common for all countries, providing a unified approach to GES assessment of two case study areas: the Adriatic Sea and the Saronikos Gulf. Dissolved Inorganic Nitrogen (DIN) threshold values of 15.6, 6.85, 1.61 and 2.11 µmol L-1 were set for the Water Types I, II, IIIW and IIIE, respectively. We also tested if an aggregation tool for GES assessment, such as Nested Environmental status Assessment Tool (NEAT), could be used as a common methodological approach. The comparison of NEAT with TRIX showed good comparability. In this end, NEAT can be used as a useful and much needed assessment tool for assessing eutrophication status of the marine.

Unveiling the ecological status of the Arabian Gulf's marine ecosystem: Insights from benthic community analysis.

This study assessed the ecological health of waters within the Saudi Arabian Exclusive Economic Zone, by utilizing benthic biotic indices with a marine monitoring dataset covering the years 2013 to 2018. This comprehensive evaluation covered a vast expanse, encompassing 67 distinctive sampling locations characterized by a wide range of depth and salinity gradients. The study examined spatial fluctuations in the benthic community and assessed potential correlations with environmental variables, including salinity, depth, sediment texture, total organic carbon, and other relevant factors. The macrobenthic density varied across the study sites, with an average density of 566 ± 120 ind.m-2. The Shannon diversity index ranged from 3.21 and 5.90, with an average of 4.70 ± 0.52. Based on the average AMBI values, all the locations were categorized as either slightly disturbed or undisturbed. Additionally, the M-AMBI analysis indicated that 95.5 % sites were in good or high ecological status.

Cross-basin and cross-taxa patterns of marine community tropicalization and deborealization in warming European seas.

Ocean warming and acidification, decreases in dissolved oxygen concentrations, and changes in primary production are causing an unprecedented global redistribution of marine life. The identification of underlying ecological processes underpinning marine species turnover, particularly the prevalence of increases of warm-water species or declines of cold-water species, has been recently debated in the context of ocean warming. Here, we track changes in the mean thermal affinity of marine communities across European seas by calculating the Community Temperature Index for 65 biodiversity time series collected over four decades and containing 1,817 species from different communities (zooplankton, coastal benthos, pelagic and demersal invertebrates and fish). We show that most communities and sites have clearly responded to ongoing ocean warming via abundance increases of warm-water species (tropicalization, 54%) and decreases of cold-water species (deborealization, 18%). Tropicalization dominated Atlantic sites compared to semi-enclosed basins such as the Mediterranean and Baltic Seas, probably due to physical barrier constraints to connectivity and species colonization. Semi-enclosed basins appeared to be particularly vulnerable to ocean warming, experiencing the fastest rates of warming and biodiversity loss through deborealization.

A dataset and template for assessing the ecological status of marine sediments and waters, based on microbial taxa.

Microbes have often been overlooked as indicators of how the ecological status is affected by human pressures. Recently, the biotic index microgAMBI was proposed to assess the status of marine sediments and waters, and it has been tested under different pressures and biogeographical areas. This index is based on the assignation of microbial taxa to one of two ecological groups: sensitive or tolerant to pollution or disturbance. The resulting taxa list has grown significantly since its first publication. Given the growing use of microgAMBI, it is crucial to make it more FAIR: Findable, Accessible, Interoperable and Reusable. Hence, this work provides the calculation template, the updated taxa list (1,974 taxa currently), and instructions on how to access and use them for assessing marine microbial ecological status.

Impacts of human activities on the supply of marine ecosystem services: A conceptual model for offshore wind farms to aid quantitative assessments.

Increased pressures from human activities may cause cumulative ecological effects on marine ecosystems. Increasingly, the study of ecosystem services is applied in the marine environment to assess the full effects of human activities on the ecosystem and on the benefits it provides. However, in the marine environment, such integrated studies have yet to move from qualitative and score-based to fully quantitative assessments. To bridge this gap, this study proposed a 4-tiered method for summarizing available knowledge and modelling tools to aid in quantitative assessments of ecosystem services supply. First, the ecosystem functioning mechanisms underlying the supply of services are conceptually mapped. Second, the impacts of the human activity of interest are summarized and linked to the first conceptual model in a case-specific model of ecosystem services supply. Third, indicators are selected that would best represent changes in the most important parameters of the conceptual model in a quantitative manner. Fourth, the knowledge gained in the previous steps is used to select models that are most useful to quantify changes in ecosystem services supply under the human pressure of interest. This approach was applied to the case study of offshore wind energy in the Belgian part of the North Sea, which is one of the most rapidly expanding industries in the marine environment globally. This study provides a useful tool to proceed towards quantification of marine ecosystem services, highlighting the need for a fully integrated approach to developing environmental impact assessment tools.

Assessing the ecological quality status of macrobenthic communities in a marine terminal of liquefied natural gas in Peru.

Macrobenthic organisms are useful bioindicators to assess ecological quality status. On the south-central coast of Peru (13°15.15'S, 76°18.5'W), a Liquefied Natural Gas (LNG) marine terminal has been operating since 2010. We investigated the macrobenthic communities and sediment parameters from 2011 to 2020 to evaluate the ecological quality status in the surrounding area of the marine terminal, using the AZTI Marine Biotic Index (AMBI) and its multivariate version (M-AMBI). We analyzed the diversity and community composition of macrobenthic invertebrates and the physico-chemical parameters of the sediment from 29 sampling sites, ranging from 0 to 15m depth. The sampling design considered: the direct influence zone ("DIZ", surroundings of the marine terminal), and northern (NCZ) and southern (SCZ) control zones. Our results indicated that abundance was high at SCZ and decreased with depth. Species richness and diversity were high at DIZ and NCZ, respectively, and increased up to 10m but dropped at 15m. High sand content was recorded in shallow depths, while in deeper areas and DIZ, mud and organic matter increased and redox potential was negative. AMBI indicated a "slightly disturbed" status in general, while M-AMBI indicated "good" or "moderate" status at depths ≤ 12m, and "poor" status at 15m. Overall, the season/year factor was not important, and variables were mostly significantly different across depths. Redox potential and organic matter were correlated with M-AMBI at 15m. In general, our results indicate an acceptable ecological quality surrounding the marine terminal, likely because the study area is not influenced by an important input of an anthropogenic stressor. This study highlights the importance of monitoring benthic communities in the surroundings of human-made structures and the use of ecological quality indices for understanding potential impacts.

Perceived multiple stressor effects depend on sample size and stressor gradient length.

Multiple stressors are continuously deteriorating surface waters worldwide, posing many challenges for their conservation and restoration. Combined effect types of multiple stressors range from single-stressor dominance to complex interactions. Identifying prevalent combined effect types is critical for environmental management, as it helps to prioritise key stressors for mitigation. However, it remains unclear whether observed single and combined stressor effects reflect true ecological processes unbiased by sample size and length of stressor gradients. Therefore, we examined the role of sample size and stressor gradient lengths in 158 paired-stressor response cases with over 120,000 samples from rivers, lakes, transitional and marine ecosystems around the world. For each case, we split the overall stressor gradient into two partial gradients (lower and upper) and investigated associated changes in single and combined stressor effects. Sample size influenced the identified combined effect types, and stressor interactions were less likely for cases with fewer samples. After splitting gradients, 40 % of cases showed a change in combined effect type, 30 % no change, and 31 % showed a loss in stressor effects. These findings suggest that identified combined effect types may often be statistical artefacts rather than representing ecological processes. In 58 % of cases, we observed changes in stressor effect directions after the gradient split, suggesting unimodal stressor effects. In general, such non-linear responses were more pronounced for organisms at higher trophic levels. We conclude that observed multiple stressor effects are not solely determined by ecological processes, but also strongly depend on sampling design. Observed effects are likely to change when sample size and/or gradient length are modified. Our study highlights the need for improved monitoring programmes with sufficient sample size and stressor gradient coverage. Our findings emphasize the importance of adaptive management, as stress reduction measures or further ecosystem degradation may change multiple stressor-effect relationships, which will then require associated changes in management strategies.

Antibiotics in the Basque coast (N Spain): Occurrence in waste and receiving waters, and risk assessment (2017-2020).

The study of the presence of antibiotics in the aquatic environment is a preliminary step to analyse their possible harmful effects on aquatic ecosystems. In order to monitor their occurrence in the aquatic environment, the European Commission established in 2015, 2018, and 2020 three Watch Lists of substances for Union-wide monitoring (Decisions (EU) 2015/495, 2018/840, and 2020/1161), where some antibiotics within the classes of macrolides, fluoroquinolones and penicillins were included. In the Basque coast, northern Spain, three macrolide antibiotics (erythromycin, clarithromycin, azithromycin) and ciprofloxacin were monitored quarterly from 2017 to 2020 (covering a period before and after the COVID19 outbreak), in water samples collected from two Waste Water Treatment Plants (WWTPs), and three control points associated with receiving waters (transitional and coastal water bodies). This work was undertaken for the Basque Water Agency (URA). The three macrolide antibiotics in water showed a frequency of quantification >65 % in the Basque coast, with higher concentrations in the WWTP emission stations than in receiving waters. Their frequency of quantification decreased from 2017 to 2020, as did the consumption of antibiotics in Spanish primary care since 2015. Ciprofloxacin showed higher frequencies of quantification in receiving waters than in wastewaters, but the highest concentrations were observed in the WWTP emission stations. Although consumption of fluoroquinolones (among which is ciprofloxacin) in primary care in the Basque Country has decreased in recent years, this trend was not observed in the waters sampled in the present study. On the other hand, concentrations of clarithromycin, azithromycin, and ciprofloxacin in receiving waters exceeded their respective Predicted No-Effect Concentrations, so they could pose an environmental risk. These substances are widely used in human and animal medicine, so, although only ciprofloxacin is included in the third Watch List, it would be advisable to continue monitoring macrolides in the Basque coast as well.

A Bayesian Network model to identify suitable areas for offshore wave energy farms, in the framework of ecosystem approach to marine spatial planning.

The production of energy from waves is gaining attention. In its expansion strategy, technical, environmental and socioeconomic aspects should be taken into account to identify suitable areas for development of wave energy projects. In this research we provide a novel approach for suitable site identification for wave energy farms. To achieve this objective, we (i) developed a conceptual framework, considering technical, environmental and conflicts for space aspects that play a role on the development of those projects, and (ii) it was operationalized in a Bayesian Network, by building a spatially explicit model adopting the Spanish and Portuguese Economic Exclusive Zones as case study. The model results indicate that 1723 km2 and 17,409 km2 are highly suitable or suitable for the development of wave energy projects (i.e. low potential conflicts with other activities and low ecological risk). Suitable areas account for a total of 2.5 TWh∙m-1 energy resource. These areas are placed between 82 and 111 m water depth, 18-30 km to the nearest port, 21-29 km to the nearest electrical substation onshore, with 143-170 MWh m-1 mean annual energy resource and having 124-150 of good weather windows per year for construction and maintenance work. The approach proposed supports scientists, managers and industry, reducing uncertainties during the consenting process, by identifying the most relevant technical, environmental and socioeconomic factors when authorising wave energy projects. The model and the suitability maps produced can be used during site identification processes, informing Strategic Environmental Assessment and ecosystem approach to marine spatial planning.

Surfing the waves: Environmental and socio-economic aspects of surf tourism and recreation.

Marine ecosystems contribute to human well-being, e.g. through the promotion of nature-based recreational activities such as surfing, which is a benefit obtained from Cultural Ecosystem Services (CES). Our research objective is to identify the benefits and impacts associated to surfing, and who are the main affected subjects and/or objects, achieving a better understanding of the sustainability status of this recreational activity. To this end, a bibliometric study and systematic review was carried out for the period 1965-2021. Benefits and impacts were collated and grouped according to their dimensional focus and type of effects in 6 groups (3-dimensional focus × 2 type of effects). The results revealed that since the beginning of 21st century surfing research topics are growing and diversifying. This review shows that implications of surfing go beyond direct users (i.e., surfers) and has consequences in diverse dimensions (environmental, socio cultural and economic), involving many stakeholders (e.g., scientific, and local communities). Most of the pieces of evidence collated in this research were related with the people who practice the activity and its social implications (psychological benefits as main benefit and injuries as main impact). Following an interdisciplinary approach, we obtained a holistic understanding of the surfing activity, not only in terms of the different dimensions addressed but on the sectors of the society that obtain benefits or are impacted by the activity. All of them should be considered and integrated to guarantee the sustainable management of this CES benefit.

An integrated assessment of the Good Environmental Status of Mediterranean Marine Protected Areas.

Local, regional and global targets have been set to halt marine biodiversity loss. Europe has set its own policy targets to achieve Good Environmental Status (GES) of marine ecosystems by implementing the Marine Strategy Framework Directive (MSFD) across member states. We combined an extensive dataset across five Mediterranean ecoregions including 26 Marine Protected Areas (MPAs), their reference unprotected areas, and a no-trawl case study. Our aim was to assess if MPAs reach GES, if their effects are local or can be detected at ecoregion level or up to a Mediterranean scale, and which are the ecosystem components driving GES achievement. This was undertaken by using the analytical tool NEAT (Nested Environmental status Assessment Tool), which allows an integrated assessment of the status of marine systems. We adopted an ecosystem approach by integrating data from several ecosystem components: the seagrass Posidonia oceanica, macroalgae, sea urchins and fish. Thresholds to define the GES were set by dedicated workshops and literature review. In the Western Mediterranean, most MPAs are in good/high status, with P. oceanica and fish driving this result within MPAs. However, GES is achieved only at a local level, and the Mediterranean Sea, as a whole, results in a moderate environmental status. Macroalgal forests are overall in bad condition, confirming their status at risk. The results are significantly affected by the assumption that discrete observations over small spatial scales are representative of the total extension investigated. This calls for large-scale, dedicated assessments to realistically detect environmental status changes under different conditions. Understanding MPAs effectiveness in reaching GES is crucial to assess their role as sentinel observatories of marine systems. MPAs and trawling bans can locally contribute to the attainment of GES and to the fulfillment of the MSFD objectives. Building confidence in setting thresholds between GES and non-GES, investing in long-term monitoring, increasing the spatial extent of sampling areas, rethinking and broadening the scope of complementary tools of protection (e.g., Natura 2000 Sites), are indicated as solutions to ameliorate the status of the basin.

Microbial diversity alteration reveals biomarkers of contamination in soil-river-lake continuum.

Microbial communities inhabiting soil-water-sediment continuum in coastal areas provide important ecosystem services. Their adaptation in response to environmental stressors, particularly mitigating the impact of pollutants discharged from human activities, has been considered for the development of microbial biomonitoring tools, but their use is still in the infancy. Here, chemical and molecular (16S rRNA gene metabarcoding) approaches were combined in order to determine the impact of pollutants on microbial assemblages inhabiting the aquatic network of a soil-water-sediment continuum around the Ichkeul Lake (Tunisia), an area highly impacted by human activities. Samples were collected within the soil-river-lake continuum at three stations in dry (summer) and wet (winter) seasons. The contaminant pressure index (PI), which integrates Polycyclic aromatic hydrocarbons (PAHs), alkanes, Organochlorine pesticides (OCPs) and metal contents, and the microbial pressure index microgAMBI, based on bacterial community structure, showed significant correlation with contamination level and differences between seasons. The comparison of prokaryotic communities further revealed specific assemblages for soil, river and lake sediments. Correlation analyses identified potential "specialist" genera for the different compartments, whose abundances were correlated with the pollutant type found. Additionally, PICRUSt analysis revealed the metabolic potential for pollutant transformation or degradation of the identified "specialist" species, providing information to estimate the recovery capacity of the ecosystem. Such findings offer the possibility to define a relevant set of microbial indicators for assessing the effects of human activities on aquatic ecosystems. Microbial indicators, including the detection of "specialist" and sensitive taxa, and their functional capacity, might be useful, in combination with integrative microbial indices, to constitute accurate biomonitoring tools for the management and restoration of complex coastal aquatic systems.

Climate regime shifts and biodiversity redistribution in the Bay of Biscay.

Global ocean warming, wave extreme events, and accelerating sea-level rise are challenges that coastal communities must address to anticipate damages in coming decades. The objective of this study is to undertake a time-series analysis of climate change (CC) indicators within the Bay of Biscay, including the Basque coast. We used an integrated and flexible methodology, based on Generalized Additive Mixed Models, to detect trends on 19 indicators (including marine physics, chemistry, atmosphere, hydrology, geomorphology, biodiversity, and commercial species). The results of 87 long-term time series analysed (~512,000 observations), in the last four decades, indicate four groups of climate regime shifts: 1) A gradual shift associated with CC starting in the 1980s, with a warming of the sea surface down to 100 m depth in the bay (0.10-0.25 °C per decade), increase in air temperature and insolation. This warming may have impacted on benthic community redistribution in the Basque coast, favouring warm-water species relative to cold-water species. Weight at age for anchovy and sardine decreased in the last two decades. 2) Deepening of the winter mixed layer depth in the south-eastern bay that probably led to increases in nutrients, surface oxygen, and chlorophyll concentration. Current increases on chlorophyll and zooplankton (i.e., copepods) biomass are contrary to those expected under CC scenarios in the region. 3) Sea-level rise (1.5-3.5 cm per decade since 1990s), associated with CC. 4) Increase of extreme wave height events of 16.8 cm per decade in the south-eastern bay, probably related to stormy conditions in the last decade, with impacts on beach erosion. Estimating accurate rates of sea warming, sea-level rise, extreme events, and foreseeing the future pathways of marine productivity, are key to define the best adaptation measures to minimize negative CC impacts in the region.

Responses of the benthic environment to reduction in anthropogenic nutrient loading in the Seto Inland Sea (Japan), based on M-AMBI assessment.

Deterioration of the sediment environment and benthic ecosystem is an undesirable effect of eutrophication, but little is known about the response of macrobenthic communities to eutrophication and their long-term recovery. In the present study, temporal changes in benthic ecological status, associated with reductions in anthropogenic impacts on a largest semi-enclosed sea in Japan, were determined using long-term monitoring data from water and sediment quality based on the multivariate AZTI Marine Biotic Index (M-AMBI), focusing on spatial differences in anthropogenic impacts. Several sub-areas were classified based on Chlorophyll a (Chl.a) concentrations in surface water during the 1980s. Chl.a concentrations decreased in all sub-areas except the sub-area with <2 µg Chl.a L-1 from the 1990s-2010s. On the other hand, total organic carbon contents in sediment decreased and M-AMBI values increased in all sub-areas during this period may be due to reduced lateral organic matter advection from surrounding areas.

From an economic crisis to a pandemic crisis: The need for accurate marine monitoring data to take informed management decisions.

It is axomatic that a system cannot be managed unless it is measured and that the measurements occur in a rigorous, defendable manner covering relevant spatial and temporal scales. Furthermore, it is not possible to predict the future direction of a system unless any predictive approach or model is supported by empirical evidence from monitoring. The marine system is no different from any other system in these regards. This review indicates the nature and topics of marine monitoring, its constraints in times of economic austerity, the sequence of topics subject to monitoring and the amount of monitoring of various topics carried out as indicated by the number of publications and researchers. We discuss the way in which the nature of monitoring is decided and we use examples to comment on the way monitoring leads to and responds to marine management and governance.

A baseline quantitative assessment of deep-sea benthic fauna of the Gulf of Aqaba (Northern Saudi Arabia, Red Sea).

The Gulf of Aqaba (hereafter 'the Gulf') is a narrow, semi-enclosed, warm, high saline, and oligotrophic water body. This baseline study provides the first quantitative data on deep-sea (207-1281 m depth) benthos of the Gulf. Fifty-five benthic species (predominantly polychaetes) with a density of 160-670 ind. m-2, species richness of 11-25, and Shannon-Wiener diversity (H') of 3.14-4.17 bits. ind.-1 were recorded from nine stations. The density and H' of benthos of the Gulf are comparable with those of the Red Sea, while both are lower than those reported from the Arabian Sea and the Mediterranean Sea. The good-high ecological status of benthic communities indicates the absence of major stress in the deep-sea habitats of the Gulf. As large-scale urbanization is proposed in the Saudi coastal areas of the Gulf, this study is expected to provide a baseline dataset for future environmental impact assessments.

A microbial mandala for environmental monitoring: Predicting multiple impacts on estuarine prokaryote communities of the Bay of Biscay.

Routine monitoring of benthic biodiversity is critical for managing and understanding the anthropogenic impacts on marine, transitional and freshwater ecosystems. However, traditional reliance on morphological identification generally makes it cost-prohibitive to increase the scale of monitoring programmes. Metabarcoding of environmental DNA has clear potential to overcome many of the problems associated with traditional monitoring, with prokaryotes and other microorganisms showing particular promise as bioindicators. However, due to the limited knowledge regarding the ecological roles and responses of environmental microorganisms to different types of pressure, the use of de novo approaches is necessary. Here, we use two such approaches for the prediction of multiple impacts present in estuaries and coastal areas of the Bay of Biscay based on microbial communities. The first (Random Forests) is a machine learning method while the second (Threshold Indicator Taxa Analysis and quantile regression splines) is based on de novo identification of bioindicators. Our results show that both methods overlap considerably in the indicator taxa identified, but less for sequence variants. Both methods also perform well in spite of the complexity of the studied ecosystem, providing predictive models with strong correlation to reference values and fair to good agreement with ecological status groups. The ability to predict several specific types of pressure is especially appealing. The cross-validated models and biotic indices developed can be directly applied to predict the environmental status of estuaries in the same geographical region, although more work is needed to evaluate and improve them for use in new regions or habitats.

Contaminants of emerging concern in the Basque coast (N Spain): Occurrence and risk assessment for a better monitoring and management decisions.

The study of the presence in the aquatic environment of certain substances considered as contaminants of emerging concern (CEC) is a preliminary step to the analysis of the possible harmful effects on aquatic ecosystems and the establishment of the corresponding environmental quality standards. In order to monitor the occurrence of CECs in the aquatic environment, the European Commission established in 2015 and 2018 two watch-list of substances for Union-wide monitoring in the field of water policy (Decision (EU) 2015/495 and Decision (EU) 2018/840). In the coast of the Basque Country, southeast of the Bay of Biscay, 19 of these watch list substances were monitored quarterly from May 2017 to March 2019. Water samples were collected at the effluent of three wastewater treatment plants and five control points associated with receiving waters (transitional and coastal water bodies). The most frequently quantified substances were azithromycin (91%), imidacloprid (82%), clarithromycin (80%), diclofenac (78%) and erythromycin (73%), with frequencies of quantification higher in wastewaters (83-100%) than in receiving waters (70-85%). In general, concentrations in wastewater were also higher than in receiving waters, indicating a dilution effect in the environment. In receiving waters, six out of the nineteen substances monitored exceeded their respective Predicted No-Effect Concentrations: azithromycin (34%), imidacloprid (9%), 17ß-estradiol (E2) (9%), clarithromycin (7%), ciprofloxacin (7%), and diclofenac (5%); and therefore, their levels could pose an environmental risk.