Occurrence, ecological risk assessment and prioritization of pharmaceuticals and abuse drugs in estuarine waters along the São Paulo coast, Brazil.

The pollution of the surface waters by pharmaceuticals and personal care products (PPCPs) has attracted worldwide attention, but data regarding their occurrence and potential risks for the aquatic biota on tropical coastal rivers of South America are still scarce. In this context, the occurrence and the preliminary ecological risk assessment of eleven pharmaceuticals of various therapeutic classes (including cocaine and its primary metabolite, benzoylecgonine) were investigated, for the first time, in five rivers of São Paulo, southeast Brazil, covering a coastline of about 140 km, namely Perequê River, Itinga River, Mongaguá River, Itanhaém River and Guaraú River. Although these five rivers are born in well-preserved areas of the Atlantic rainforest biome, on its way to sea and when they cross the urban perimeter, they receive untreated sewage discharges containing a complex mixture of contaminants. In addition, a "persistence, bioaccumulation and toxicity" (PBT) approach allowed to pre-select the priority PPCPs to be monitored in this coastline. Identification of several PPCPs in the samples was done using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Ten PPCPs were successfully quantified in all five rivers, namely caffeine (9.00-560.00 ng/L), acetaminophen (

First report on the occurrence of pharmaceuticals and cocaine in the coastal waters of Santa Catarina, Brazil, and its related ecological risk assessment.

The worldwide occurrence of pharmaceuticals and personal care products (PPCPs) in aquatic ecosystems is reason for public concern. These emerging micropollutants include a large and diverse group of organic compounds, with continuous input, high environmental persistence and potential threat to biota and human health. The aim of this study was to evaluate, for the first time, the occurrence of twenty-seven PPCPs of various therapeutic classes (including cocaine and its primary metabolite, benzoylecgonine), in the coastal waters of Santa Catarina, southern Brazil. Water samples were taken in November 2020, during the low tide periods, at eight sampling points located along the coast of Santa Catarina, covering its entire geographical extension. Sampling was carried out in triplicate and at different depths of the water column. Nine compounds were detected through liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS): caffeine (12.58-119.80 ng/L), diclofenac (1.34-7.92 ng/L), atenolol (1.13-2.50 ng/L), losartan (0.43-3.20 ng/L), acetaminophen (0.21-10.04 ng/L), orphenadrine (0.07-0.09 ng/L), cocaine (0.02-0.17 ng/L), benzoylecgonine (0.01-1.1 ng/L) and carbamazepine (0.02-0.27 ng/L). The highest occurrence of these compounds was detected in the northern and central coastal region of Santa Catarina, namely in Penha and Palhoça cities. Moreover, the risk assessment showed that almost compounds (atenolol, benzoylecgonine, carbamazepine, cocaine and orphenadrine) presented no ecological risk in the recorded concentrations. However, a few compounds suggest low (caffeine and diclofenac) to moderate (acetaminophen and losartan) risk taking into consideration the acute and chronic effects for the three trophic levels (algae, crustacean and fish) tested. These compounds are usually found in areas with high population density, aggravated by tourism, because of the sanitary sewage and solid waste. Although in low concentrations, the occurrence of these chemical compounds can imply deleterious effects on the environmental health of Santa Catarina coastal zone, and therefore deserve more attention by the public authorities and environmental agencies.

Occurrence of pharmaceuticals and cocaine in the urban drainage channels located on the outskirts of the São Vicente Island (São Paulo, Brazil) and related ecological risk assessment.

"Wealth by the sea and poverty away from the sea breeze" is a metaphor that mirrors what happens along the Brazilian coastal zone, namely in São Vicente Island, São Paulo, Brazil. Due to the high cost of the properties on this shore, the impoverished population started to migrate to the northern outskirts of the island (away from the tourist beaches), potentiating the emergence of poor housing conditions, namely stilt-house slums. Consequently, the urban drainage channels across these outskirts neighbourhoods are potentially contaminated by human wastes. In this context, the occurrence and preliminary ecological risk assessment of eleven pharmaceuticals of various therapeutic classes (including cocaine and its primary metabolite, benzoylecgonine) were investigated, for the first time, in five urban drainage channels whose diffuse loads flow continuously to the estuarine waters of São Vicente Island. The results showed the widespread presence of these environmental stressors in all urban channels analysed, namely losartan (7.3-2680.0 ng/L), caffeine (314.0-726.0 ng/L), acetaminophen (7.0-78.2 ng/L), atenolol (6.2-23.6 ng/L), benzoylecgonine (10.2-17.2 ng/L), furosemide (1.0-7.2 ng/L), cocaine (2.3-6.7 ng/L), carbamazepine (0.2-2.6 ng/L), diclofenac (1.1-2.5 ng/L), orphenadrine (0.2-1.1 ng/L) and chlortalidone (0.5-1.0 ng/L). The overall total estimated load of pharmaceuticals and personal care products flowing to the estuarine waters of São Vicente Island is on the order of 41.1 g/day. The ecological risk assessment revealed a great environmental concern for São Vicente Island, ranging between low (e.g. carbamazepine and cocaine) and moderate to high (e.g. caffeine, acetaminophen and losartan) threats for the aquatic biota. Therefore, initiatives promoting basic sanitation, land-use regularisation and population awareness are highly recommended.

Occurrence and ecological risk assessment of pharmaceuticals and cocaine in the urban drainage channels of Santos beaches (São Paulo, Brazil): a neglected, but sensitive issue.

In some Brazilian coastal cities, it is common to observe 'black tongues' in beaches, i.e. a mixture of urban runoff and untreated domestic sewage containing pollutants of emerging concern, namely pharmaceutical and personal care products (PPCPs), flowing into the South Atlantic Ocean. Such diffuse loads of pollutants might expose nontarget aquatic organisms to harmful compounds. In this work, the occurrence and preliminary ecological risk of 27 PPCPs of various therapeutic classes (including cocaine and its primary metabolite, benzoylecgonine) were investigated, for the first time, in seven urban drainage channels whose diffuse loads flow continuously to the beaches of Santos Bay, São Paulo, Brazil. Of these, 21 compounds were detected using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS), and nine of them were consistently quantified in all urban channels of Santos, suggesting that those pollutants are ubiquitous in this region: caffeine (143.4-516.0 ng/L), losartan (4.2-21.8 ng/L), atenolol (1.1-18.2 ng/L), acetaminophen (1.5-13.8 ng/L), benzoylecgonine (1.0-4.8 ng/L), carbamazepine (1.1-4.0 ng/L), diclofenac (1.9-3.5 ng/L), cocaine (0.5-1.7 ng/L), and orphenadrine (0.1-0.8 ng/L). Moreover, twelve compounds were found below the limit of quantification (

Mutagenic and ecotoxicological assessment of urban surface runoff flowing to the beaches of Guarujá, State of São Paulo, Brazil.

Along the coast of the State of São Paulo, Brazil, urban drainage channels introduce a complex mixture of pollutants into the South Atlantic Ocean, that may cause deleterious effects to the aquatic biota. The objective of this study was to analyse, for the first time, the mutagenicity (Ames Salmonella/microsome test) and ecotoxicity (acute and chronic tests, with Daphnia simillis and Ceriodaphnia dubia, respectively) exerted by the diffuse loads discharged in Guarujá, São Paulo coast, Brazil. Water sampling occurred bimonthly between January and July 2018 (rainy season: January through March; dry season: May through July) at four beaches with different profiles of use and land occupation: Tombo (Blue Flag certification), Enseada (high use by tourists), Perequê (fishing community) and Iporanga (conservation unit). No mutagenic potential was detected in the complex mixtures flowing to the study beaches. However, 30 and 80% of the analyses showed acute and chronic toxicities, respectively, mainly in the Enseada and Perequê channels during the rainy season. To improve the environmental quality of these coastal waters and to reduce the ecological risks posed to the aquatic organisms and public health, several actions are imperative, such as the amelioration of the basic sanitation facilities and land regularisation actions.

Occurrence and risk assessment of pharmaceuticals and cocaine around the coastal submarine sewage outfall in Guarujá, São Paulo State, Brazil.

The aim of this study was to screen and quantify 23 pharmaceutical compounds (including illicit drugs), at two sampling points near the diffusers of the Guarujá submarine outfall, State of São Paulo, Brazil. Samples were collected in triplicate during the high (January 2018) and low (April 2018) seasons at two different water column depths (surface and bottom). A total of 10 compounds were detected using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Caffeine (42.3-141.0 ng/L), diclofenac (3.6-85.7 ng/L), valsartan (4.7-14.3 ng/L), benzoylecgonine (0.3-1.7 ng/L), and cocaine (0.3-0.6 ng/L) were frequently detected (75% occurrence). Orphenadrine (0.6-3.0 ng/L) and atenolol (0.1-0.3 ng/L), and acetaminophen (1.2-1.4 ng/L) and losartan (0.7-3.4 ng/L), were detected in 50% and 25% of the samples, respectively. Only one sample (12.5%) detected the presence of carbamazepine (< 0.001-0.1 ng/L). Unexpectedly a lower frequency of occurrence and concentration of these compounds occurred during the summer season, suggesting that other factors, such as the oceanographic and hydrodynamic regimes of the study area, besides the population rise, should be taken into account. Caffeine presented concentrations above the surface water safety limits (0.01 µg/L). For almost all compounds, the observed concentrations indicate nonenvironmental risk for the aquatic biota, except for caffeine, diclofenac, and acetaminophen that showed low to moderate ecological risk for the three trophic levels tested.

Occurrence and ecological risk assessment of pharmaceuticals and cocaine in a beach area of Guarujá, São Paulo State, Brazil, under the influence of urban surface runoff.

The occurrence of pharmaceuticals and illicit drugs in water resources is widely documented in Europe, North America and Asia. However, in South America, these studies are still incipient. The objective of this study was to screen and identify the presence of pharmaceuticals of various therapeutic classes, including illicit drugs such as cocaine and its metabolite benzoylecgonine, in urban drainage channels that flow into the bathing waters of Guarujá city, State of São Paulo, Brazil. Moreover, the ecological potential risks to the aquatic biota were also assessed. The water samples were collected from four beaches of Guarujá in two different points: in the urban drainage channels and in the nearby coast line. A total of 16 compounds were detected using liquid chromatography coupled with tandem mass spectrometry: carbamazepine (0.1-8.0 ng/L), caffeine (33.5-6550.0 ng/L), cocaine (0.2-30.3 ng/L), benzoylecgonine (0.9-278.0 ng/L), citalopram (0.2-0.4 ng/L), acetaminophen (18.3-391.0 ng/L), diclofenac (0.9-79.8 ng/L), orphenadrine (0.2-1.5 ng/L), atenolol (0.1-140.0 ng/L), propranolol (limit of detection: LOD-0.9 ng/L), enalapril (2.2-3.8 ng/L), losartan (3.6-548.0 ng/L), valsartan (19.8-798.0 ng/L), rosuvastatin (2.5-38.5 ng/L), chlortalidone (0.1-0.4 ng/L) and clopidogrel (0.1-0.2 ng/L). The hereby data also showed that five of these compounds, namely caffeine, acetaminophen, diclofenac, losartan and valsartan, could raise moderate to severe risks to aquatic organisms (algae, crustaceans and fishes). This study is the first report of the occurrence of several pharmaceuticals and illicit drugs in urban drainage channels that flow to the bathing waters in South America, and it is the first quantification of rosuvastatin, chlortalidone and clopidogrel in environmental marine waters of Latin America.

Occurrence of human adenoviruses in a beach area of Guarujá, São Paulo, Brazil.

Along the coastal zone of the State of São Paulo, Brazil, diffuse discharges that flow directly to tourist beaches are responsible for introducing various pathogens into recreational waters. The objective of this study was to analyze, for the first time, the presence of protozoa (Cryptosporidium ssp and Giardia ssp), as well as human mastadenoviruses (HAdV-species C and F) and other species of adenoviruses (AdV) in beach drainage channels of Enseada and Perequê, municipality of Guarujá, São Paulo, Brazil. Protozoa were not detected in any sample over the course of the 11-month study. In relation to HAdVs, 100% (n = 22) of the water samples presented contamination by at least one type of virus (C, D, or F species), suggesting potential risks to the public health following recreational exposure of beach users to these waters. PRACTITIONER POINTS: First report on the presence of human adenoviruses in urban drainage channels in the coast of São Paulo State, Brazil. Urban surface water runoff is responsible for introducing human adenoviruses linked to disease outbreaks in areas of intense recreation. Cryptosporidium oocysts and Giardia cysts were below the limit of detection for all analyzed samples. However, all sites were positive for at least one of the viral species (HAdV-C, HAdV-D, or HAdV-F). Viral loads found in the water were similar to those commonly found in the wastewater.

Seasonal monitoring of cocaine and benzoylecgonine in a subtropical coastal zone (Santos Bay, Brazil).

Illicit drugs and their metabolites represent a new class of emerging contaminants. These substances are continuously discharged into wastewater which have been detected in the aquatic environment in concentrations ranging from ng.L-1 to µg.L-1. Our study detected the occurrence of cocaine (COC) and benzoylecgonine (BE) in a subtropical coastal zone (Santos Bay, SP, Brazil) within one year. Water samples (surface and bottom) were collected from the Santos Submarine Sewage Outfall (SSOS) area. COC and BE were measured in the samples using ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-ESI-MS/MS). Concentrations ranged from 12.18 to 203.6 ng.L-1 (COC) and 8.20 to 38.59 ng.L-1 (BE). Higher concentrations of COC were observed during the end of spring, following the population increase at summer season. COC and its metabolite occurrence in this coastal zone represent a threat to coastal organisms.

Marine contamination and cytogenotoxic effects of fluoxetine in the tropical brown mussel Perna perna.

Concerns are growing about the presence of fluoxetine (FLX) in environmental matrices, as well as its harmful effects on non-target organisms. FLX in aquatic ecosystems has been detected in a range varying from pg/L to ng/L, while adverse effects have been reported in several organisms inhabiting freshwater and marine environments. The present study quantifies FLX concentrations in seawater samples from Santos Bay, Brazil and assesses metabolic responses and sublethal effects on the tropical brown mussel Perna perna. Levels of ethoxyresorufin­O­deethylase, dibenzylfluorescein dealkylase, glutathione S-transferase, glutathione peroxidase, cholinesterase, lipoperoxidation, and DNA damage were assessed in the gills and digestive gland of these animals, and lysosomal membrane stability was also assessed in hemocytes. FLX altered phase I and II enzyme activities, caused cytogenotoxic effects, and negatively impacted the overall health of mussels exposed to environmentally relevant concentrations. These findings contribute to characterize the risks of introducing this drug into the marine environment.

Ecotoxicological effects of losartan on the brown mussel Perna perna and its occurrence in seawater from Santos Bay (Brazil).

The antihypertensive losartan (LOS) has been detected in wastewater and environmental matrices, however further studies focused on assessing the ecotoxicological effects on aquatic ecosystems are necessary. Considering the intensive use of this pharmaceutical and its discharges into coastal zones, our study aimed to determine the environmental concentrations of LOS in seawater, as well as to assess the biological effects of LOS on the marine bivalve Perna perna. For this purpose, fertilization rate and embryolarval development were evaluated through standardized assays. Phase I (ethoxyresorufin O­deethylase EROD and dibenzylfluorescein dealkylase DBF) and II (glutathione S-transferase GST) enzymes, glutathione peroxidase (GPx), Cholinesterase (ChE), lipoperoxidation (LPO) and DNA damage were used to analyze sublethal responses in gills and digestive gland of adult individuals. Lysosomal membrane stability was also assessed in hemocytes. Our results showed the occurrence of LOS in 100% of the analyzed water samples located in Santos Bay, Sao Paulo, Brazil, in a range of 0.2 ng/L-8.7 ng/L. Effects on reproductive endpoints were observed after short-term exposure to concentrations up to 75 mg/L. Biomarker responses demonstrated the induction of CYP450 like activity and GST in mussel gills exposed to 300 and 3000 ng/L of LOS, respectively. GPx activity was also increased in concentration of exposure to 3000 ng/L of LOS. Cyto-genotoxic effects were found in gills and hemocytes exposed in concentrations up to 300 ng/L. These results highlighted the concern of introducing this class of contaminants into marine environments, and pointed out the need to include antihypertensive compounds in environmental monitoring programs.

A tiered approach to assess effects of diclofenac on the brown mussel Perna perna: A contribution to characterize the hazard.

Pharmaceutical discharges into the aquatic ecosystem are of environmental concern and sewage treatment plants (STPs) have been pointed out as the major source of these compounds to coastal zones, where oceanic disposal of sewage occurs through submarine outfalls. Diclofenac (DCF) is one of the most frequently detected pharmaceuticals in water, but little is known about the effects on marine organisms. In this study, we employed a tiered approach involving the determination of environmental concentrations of DCF in marine water and the adverse biological effects for fertilization, embryo-larval development and biomarker responses of the mussel Perna perna. Results indicate that effects in fertilization rate and embryo-larval development were found in the order of mg·L-1. However, low concentrations of DCF (ng·L-1) significantly decreased the lysosomal membrane stability and COX activity, as well as triggered DNA damage, oxidative stress and changes in antioxidant defenses. Our results point to an environmental hazard at coastal ecosystems and suggest the need for improvements in the treatment of domestic wastewater aiming to reduce DCF concentrations, as well as regulation on current environmental legislation and monitoring of aquatic ecosystems.

Protein disulfide isomerase redox-dependent association with p47(phox): evidence for an organizer role in leukocyte NADPH oxidase activation.

Mechanisms of leukocyte NADPH oxidase regulation remain actively investigated. We showed previously that vascular and macrophage oxidase complexes are regulated by the associated redox chaperone PDI. Here, we investigated the occurrence and possible underlying mechanisms of PDI-mediated regulation of neutrophil NADPH oxidase. In a semirecombinant cell-free system, PDI inhibitors scrRNase (100 µg/mL) or bacitracin (1 mM) near totally suppressed superoxide generation. Exogenously incubated, oxidized PDI increased (by ~40%), whereas PDIred diminished (by ~60%) superoxide generation. No change occurred after incubation with PDI serine-mutated in all four redox cysteines. Moreover, a mimetic CxxC PDI inhibited superoxide production by ~70%. Thus, oxidized PDI supports, whereas reduced PDI down-regulates, intrinsic membrane NADPH oxidase complex activity. In whole neutrophils, immunoprecipitation and colocalization experiments demonstrated PDI association with membrane complex subunits and prominent thiol-mediated interaction with p47(phox) in the cytosol fraction. Upon PMA stimulation, PDI was mobilized from azurophilic granules to cytosol but did not further accumulate in membranes, contrarily to p47(phox). PDI-p47(phox) association in cytosol increased concomitantly to opposite redox switches of both proteins; there was marked reductive shift of cytosol PDI and maintainance of predominantly oxidized PDI in the membrane. Pulldown assays further indicated predominant association between PDIred and p47(phox) in cytosol. Incubation of purified PDI (>80% reduced) and p47(phox) in vitro promoted their arachidonate-dependent association. Such PDI behavior is consistent with a novel cytosolic regulatory loop for oxidase complex (re)cycling. Altogether, PDI seems to exhibit a supportive effect on NADPH oxidase activity by acting as a redox-dependent enzyme complex organizer.