Computational modeling (in silico) methods combined with ecotoxicological experiments (in vivo) to predict the environmental risks of an antihistamine drug (loratadine).

This study employed computational modeling (in silico) methods, combined with ecotoxicological experiments (in vivo) to predict the persistence/biodegradability, bioaccumulation, mobility, and ecological risks of an antihistamine drug (Loratadine: LOR) in the aquatic compartment. To achieve these goals, four endpoints of the LOR were obtained from different open-source computational tools, namely: (i) "STP total removal"; (ii) Predicted ready biodegradability; (iii) Octanol-water partition coefficient (KOW); and (iv) Soil organic adsorption coefficient (KOC). Moreover, acute and chronic, ecotoxicological assays using non-target freshwater organisms of different trophic levels (namely, algae Pseudokirchneriella subcapitata; microcrustaceans Daphnia similis and Ceriodaphnia dubia; and fish Danio rerio), were used to predict the ecological risks of LOR. The main results showed that LOR: (i) is considered persistent (after a weight-of-evidence assessment) and highly resistant to biodegradation; (ii) is hydrophobic (LOG KOW = 5.20), immobile (LOG KOC = 5.63), and thus, it can potentially bioaccumulate and/or can cause numerous deleterious effects in aquatic species; and (iii) after ecotoxicological evaluation is considered "toxic" and/or "highly toxic" to the three trophic levels tested. Moreover, both the ecotoxicological assays and risk assessment (RQ), showed that LOR is more harmful for the crustaceans (RQcrustaceans = moderate to high risks) than for algae and fish. Ultimately, this study reinforces the ecological concern due to the indiscriminate disposal of this antihistamine drug in worldwide aquatic ecosystems.

Otolith shape and elemental signatures provide insights into the connectivity of euryhaline Chelon labrosus inhabiting two close estuaries with different burdens of xenoestrogens in the Southern Bay of Biscay.

Intersex gonads have been observed in thicklip grey mullet Chelon labrosus, inhabiting estuaries with high burdens of xenoestrogens in the Southeast Bay of Biscay, but knowledge of population connectivity among estuaries is lacking for this euryhaline fish species. This study investigates the population structure of C. labrosus using otolith shape and elemental signatures of 60 adults (overall length âˆ¼ 38 cm) from two estuaries 21 nautic miles apart, one with a high incidence of intersex condition (Gernika), and the other one pristine (Plentzia). Otolith shape analyses were performed using elliptical Fourier descriptors, while elemental signatures of whole sagittae were obtained by inductively coupled plasma mass spectrophotometry. Univariate and multivariate statistics were applied to determine if otolith signatures show patterns of homogeneity between estuaries. The data indicated significant differences in the otolith shape and elemental composition between mullets of Gernika and Plentzia. Elemental differences were mainly driven by Sr, Li (both higher in Plentzia) and Ba (higher in Gernika). The 98% re-classification success rate obtained from stepwise linear discriminant function analysis suggests that Gernika and Plentzia individuals form separated population units. The limited connectivity between these two close estuaries would indicate a different life history of exposure to chemicals, which might explain the high prevalence of intersex condition in Gernika and its absence in Plenztia.

Identifying Whitemouth Croaker (Micropogonias furnieri) Populations along the Rio de Janeiro Coast, Brazil, through Microsatellite and Otolith Analyses.

The inshore area of the Southwestern Atlantic between 22 °S and 29 °S (South Brazilian Bight) is a transitional climatic zone, where the tropical and warm temperate provinces mix. In its northern part, i.e., in the coastal waters of Rio de Janeiro, Brazil, local oceanographic conditions, such as upwelling in the north, and great bays with different degrees of anthropogenic influences in the center and south can determine the population structure of several fish stocks. The Whitemouth croaker (Micropogonias furnieri) is one the most heavily exploited fishing resources in this area, but there are still some doubts about its population structure. In this study, through combined analyses using nuclear genetic markers and morphological and geochemical signatures of otoliths, a divergence of individuals between two populations was identified using microsatellites, while a finer spatial structure with three populations (north, center and south, respectively) was found based on otolith shapes and elemental signatures. This regional population structure may have direct implications for rational fisheries management and conservation of the species.

Past and Contemporaneous Otolith Fingerprints Reveal Potential Anthropogenic Interferences and Allows Refinement of the Population Structure of Isopisthus parvipinnis in the South Brazil Bight.

In this study, otolith geochemical signatures (Element:Ca ratios) were used to investigate the long-term spatial shifts of the population structure of Isopisthus parvipinnis, Bigtooth corvina, an economically and ecologically important Brazilian fish species. Two-hundred and ninety-seven juvenile individuals from historical (1975) and contemporary (2018/2019) samples were collected in five subareas [São Paulo: North-NSP, Center-CSP and South-SSP; Paraná (PR) and Santa Catarina (SC)] of the shallow waters off the coast of the South Brazil Bight were analyzed. The main informative single elements were Co:Ca, Cu:Ca, Li:Ca, Mg:Ca, Mn:Ca, Ni:Ca, Na:Ca, and Rb:Ca. Multivariate analysis showed spatial differences in otolith chemical composition over the years. Samples from 1975 presented an overall low reclassification rate (58%), suggesting the existence of two population units: (1) SP + PR; and (2) SC. However, samples from 2018/2019 discriminated four distinct population units with a good overall reclassification (80%): (1) NSP; (2) CSP; (3) SSP + PR; and (4) SC. This spatial differentiation on the geochemical signatures probably reflects the effects of long-term temporal variability in oceanographic conditions, anthropogenic influence, and climate change on this coastal ecosystem. The data also corroborate and refines the population structure scenario of I. parvipinnis recently described using complementary phenotypic tags.

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 (

Assessment of Paracetamol Toxic Effects under Varying Seawater pH Conditions on the Marine Polychaete Hediste diversicolor Using Biochemical Endpoints.

Increasing atmospheric carbon dioxide (CO2) levels are likely to lower ocean pH values, after its dissolution in seawater. Additionally, pharmaceuticals drugs are environmental stressors due to their intrinsic properties and worldwide occurrence. It is thus of the utmost importance to assess the combined effects of pH decreases and pharmaceutical contamination, considering that their absorption (and effects) are likely to be strongly affected by changes in oceanic pH. To attain this goal, individuals of the marine polychaete Hediste diversicolor were exposed to distinct pH levels (8.2, 7.9, and 7.6) and environmentally relevant concentrations of the acidic drug paracetamol (PAR: 0, 30, 60, and 120 µg/L). Biomarkers such as catalase (CAT), glutathione S-transferases (GSTs), acetylcholinesterase (AChE), and cyclooxygenase (COX) activities, as well as peroxidative damage (through thiobarbituric acid reactive substance (TBARS) quantification), were quantified to serve as ecotoxicological endpoints. Data showed a general increase in CAT and a decrease in GST activities (with significant fluctuations according to the tested conditions of PAR and pH). These changes are likely to be associated with alterations of the redox cycle driven by PAR exposure. In addition, pH levels seemed to condition the toxicity caused by PAR, suggesting that the toxic effects of this drug were in some cases enhanced by more acidic conditions. An inhibition of AChE was observed in animals exposed to the highest concentration of PAR, regardless of the pH value. Moreover, no lipid peroxidation was observed in most individuals, although a significant increase in TBARS levels was observed for polychaetes exposed to the lowest pH. Finally, no alterations of COX activities were recorded on polychaetes exposed to PAR, regardless of the pH level. The obtained results suggest that seawater acidification is detrimental to marine wildlife, since it may enhance toxic effects caused by environmental realistic concentrations of acidic drugs, such as PAR. This work was crucial to evidence that ocean acidification, in the context of a global change scenario of increased levels of both atmospheric and oceanic CO2, is a key factor in understanding the putative enhanced toxicity of most pharmaceutical drugs that are of an acidic nature.

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 (

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.

Multi-biomarker approach to assess the acute effects of cerium dioxide nanoparticles in gills, liver and kidney of Oncorhynchus mykiss.

Cerium oxide nanoparticles (CeO2-NP) have already been detected in the aquatic compartment, however, the evaluation of potential ecotoxicological effects on biota are scarce. The present study aimed to assess the toxic effects of CeO2-NP in Oncorhynchus mykiss in different organs/tissues (gills, liver and kidney) after acute exposure (96 h) to three concentrations: 0.25, 2.5 and 25 mg/L. Oxidative stress response (catalase - CAT; glutathione S-transferases - GSTs), lipid peroxidation (thiobarbituric acid reactive substances - TBARS), Na+/K+-ATPase activity, genotoxicity (genetic damage index - GDI) and histopathology (organ's pathological indices) were evaluated. CAT activity was increased in gills and decreased in liver of fish exposed to the highest CeO2-NPs concentration tested. However, GSTs and Na+/K+-ATPase activities and TBARS levels were not significantly altered in analysed organs. CeO2-NP caused marked changes in the gills (aneurysms, blood capillary congestion, lamellar hypertrophy and hyperplasia, secondary lamella fusion and epithelial lifting), in liver (pyknotic nucleus, hyperemia, enlargement of sinusoids and leucocyte infiltration) and kidney (shrinkage of the glomeruli, enlargement of the Bowman space, tubular degeneration and nuclear hypertrophy). Moreover, a semi-quantitative histopathological scoring system (pathological index) confirmed significant alterations in the three organs of all exposed fish. Furthermore, a significant increase of GDI indices observed in gills and liver, for all tested concentrations, indicated a dose-dependent effect. The present study suggests that the release of CeO2-NP into the aquatic environment promotes biochemical, genotoxic and histopathological damages in fish. However, the mechanisms underlying the occurrence of such effects require further investigation.

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.

Effects of pH on salicylic acid toxicity in terms of biomarkers determined in the marine gastropod Gibbula umbilicalis.

Alterations of the physical-chemical properties of the oceans due to anthropogenic activities are, at present, one of the most concerning environmental issues studied by researchers. One of these issues is ocean acidification, mainly caused by overproduction and release of carbon dioxide (CO2) from anthropogenic sources. Another component of environmental degradation is related to the production and release of potential toxic compounds, namely active pharmaceutical ingredients, into the aquatic environment that, combined with oceanic acidification, can cause unpredictable and never before considered deleterious effects on non-target marine organisms. Regarding this issue, the hereby study used predictions of future ocean acidification to simulate realistic scenarios of environmental exposure to a common therapeutic drug, salicylic acid (SA), in the marine gastropod Gibbula umbilicalis under different pH values. This species was exposed to a range of pH values (8.2, 7.9 and 7.6), and to already reported environmentally realistic concentrations (5, 25 and 125 µg/L) of SA. To evaluate the effects of these environmental stressors, key physiological biomarkers (GSTs, CAT, TBARS, AChE and COX) and shell hardness (SH) were quantified. Results from the present study showed that CAT and GSTs activities were enhanced by SA under water acidification; increased lipid peroxidation was also observed in organisms exposed to SA in more acidic media. In addition, the hereby study demonstrated the neurotoxic effects of SA through the inhibition of AChE. Effects were also observed in terms of COX activity, showing that SA absorption may be affected by water acidification. In terms of SH, the obtained data suggest that SA may alter the physical integrity of shells of exposed organisms. It is possible to conclude that the combination of seawater acidification and exposure to toxic xenobiotics (namely to the drug SA) may be strenuous to marine communities, making aquatic biota more susceptible to xenobiotics, and consequently endangering marine life in an unpredictable extent.

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.

Testing the impact of contaminated sediments from the southeast marine coast of Tunisia on biota: a multibiomarker approach using the flatfish Solea senegalensis.

Coastal marine areas are highly vulnerable to the exposure to various types of stressors and impact of chemical pollution resulting from increasing anthropogenic activities, namely pollution by metals and polycyclic aromatic hydrocarbons (PAHs). To assess ecosystem quality and functions, biomarkers can provide information about the presence and adverse effects of pollutants. Accordingly, the present study was conducted to evaluate the chronic (28 days) biologic effects of putatively contaminated sediments from the Zarzis area, located in the south of the Gulf of Gabes on the Southern Tunisian coast, on the marine flatfish Solea senegalensis. Sediments were collected at three sampling sites, impacted by wastewater discharges, aquaculture activities, and industrial contamination, and then surveyed for metals (Cd, Cu, Cr, Hg, Zn, and Pb) and organic contaminants (polycyclic aromatic hydrocarbons). The quantified biomarkers involved the determination of oxidative stress, phase II metabolism, and the extent of lipid peroxidation (catalase, CAT; glutathione peroxidase activity: total and selenium-dependent, T-GPx and Se-GPx; activities of glutathione-S-transferases, GSTs; levels of lipid peroxidation, by means of the thiobarbituric acid reactive substances assay, TBARS) and neurotoxicity (activity of acetylcholinesterase, AChE). S. senegalensis exposed to potentially contaminated sediments, collected near the aquaculture facility, presented the highest values for the generality of biomarkers tested, and a significant inhibition of AChE activity. A few lesions have been also recorded in the gills and liver tissues of S. senegalensis following chronic exposure. However, the observed lesions in gills (e.g., epithelial lifting, lamellar fusion, gills hyperplasia and hypertrophy, and leukocyte infiltration) and liver (cytoplasmic vacuolation, enlargement of sinusoids, foci of necrosis, and eosinophilic bodies) were of minimal pathological importance and/or low prevalence that did not significantly affect the weighted histopathological indices. Finally, the biological responses evidenced by this flatfish can be potentially caused by metal and PAH pollution occurring in specific areas in the southeast of Tunisia. The type and extent of the observed biochemical alterations strongly suggest that the contaminated sediments from the surveyed areas could cause early adverse biological effects on exposed biota.

Histopathological effects of the antibiotic erythromycin on the freshwater fish species Oncorhynchus mykiss.

Pharmaceuticals are found in the aquatic compartment due to their continuous release in wastewater effluents or direct dispersal in aquaculture practices raising serious threats to human and environmental health. Erythromycin (ERY) is a macrolide antibiotic widely prescribed in human and veterinary medicine to threat a number of bacterial infections, being consequently found in the aquatic environment. The present work intends to evaluate the sub-lethal effects of ERY on juveniles of rainbow trout (Oncorhynchus mykiss) in terms of tissue damage using histochemical staining procedures. Individuals were exposed for 96 h (acute exposure: 0.001-10 mg/L) and 28 days (chronic exposure: 0.05-0.8 µg/L) to environmentally realistic concentrations of ERY. Qualitative and quantitative approaches were used to assess O. mykiss gills and liver tissue alterations after exposure to ERY. For both exposures the most common gill changes recorded were progressive (e.g. hypertrophy of mucous cells and hyperplasia of the epithelial cells). However, circulatory (e.g. aneurysms and oedemas) and regressive (e.g. epithelial lifting of lamellae and lamellar fusion) changes were also observed in the acute assay. Gill morphometric analysis revealed to be a good indicator of subtle alterations in gill architecture in agreement with the qualitative scoring system. In liver, regressive (e.g. cytoplasmic vacuolization, pyknotic nucleus and hepatocellular degeneration) and circulatory disturbances (e.g. hemorrhage and increase of sinusoidal space) were the most frequently observed alterations, but only for the acute assay. Furthermore, all histological changes observed contributed to a significant increase in the pathological index for both organs. The current data demonstrate the existence of a direct dose-effect relationship between the exposure to this specific macrolide antibiotic and the histological disorders recorded in different tissues of the exposed fish. The histopathological findings observed in this study may have been the result of several physio-metabolic dysfunctions. However, the observed tissue lesions were of minimal or moderate pathological importance, non-specific and reversible. Further investigation into the cellular mechanism of action of ERY is needed.

Histopathological effects in gills and liver of Sparus aurata following acute and chronic exposures to erythromycin and oxytetracycline.

Due to their worldwide use and environmental persistence, antibiotics are frequently detected in various aquatic compartments. Their toxic properties raise environmental concerns to non-target organisms. Histopathology data is frequently applied in ecotoxicology studies to assess the effects of different classes of environmental stressors in fish, including antibiotics. Tissue alterations in gills and liver of gilthead seabream (Sparus aurata) individuals acutely (96 h) and chronically (28 days) exposed to environmentally relevant concentrations of the antibiotics erythromycin (ERY: 0.0002-200 µg/L) and oxytetracycline (OTC: 0.0004-400 µg/L), including a control non-exposed group, were evaluated. Several disorders (circulatory, regressive, progressive, and inflammatory) were observed in both organs of all exposed animals. The hereby obtained data showed a higher and significant increase in gill histopathological index of organisms acutely exposed to ERY and of those chronically exposed to OTC. In terms of categorical lesions, only a significant increase of regressive and progressive alterations occurred in gills after chronic exposure to OTC. For the liver, a significant increase in pathological index was also detected, as well as regressive changes, after chronic exposure to OTC. Furthermore, the present study indicates that most of the changes observed in gills and liver were of mild to moderate severity, which might be adaptive or protective, non-specific, and mostly reversible. Despite being observed, irreversible lesions were not significant in any of the fish organs analyzed. Although there were histological changes, gill apparatus was considered still functionally normal, as well as liver tissue, not supporting the occurrence of severe toxicity. In general, the observed histological changes were not stressor-specific, and toxicological mechanistic explanations for the alterations observed in gills and liver are presented. The obtained data showed that histopathological biomarkers can be successfully applied in ecotoxicological studies, evidencing their relevance, responsivity, and complementarity to other biochemical biomarker-based approaches.

Worms on drugs: ecotoxicological effects of acetylsalicylic acid on the Polychaeta species Hediste diversicolor in terms of biochemical and histological alterations.

Pharmaceuticals are important environmental stressors since they have a worldwide use; they are usually released in the aquatic compartment without adequate treatment, and because of their intrinsic properties, they may affect several non-target organisms. Acetylsalicylic acid (ASA), the active substance of aspirin, is a non-steroidal anti-inflammatory drug, being one of the most widely prescribed analgesics in human medical care. Consequently, this compound is systematically reported to occur in the wild, where it may exert toxic effects on non-target species, which are mostly uncharacterized so far. The objective of the present work was to assess the acute and chronic effects of ASA on selected oxidative stress biomarkers [catalase (CAT), glutathione reductase (GRed), glutathione peroxidase (GPx), glutathione S-transferase (GST)], lipid peroxidation (thiobarbituric acid-reactive substance), and histological alterations in the polychaete Hediste diversicolor (Annelida: Polychaeta). The obtained data showed that ASA is not exempt of toxicity, since it was responsible for significant, albeit transient, changes in biomarkers related to the redox status of the organisms, occurring as an increase in the activity of catalase in the individuals exposed acutely to ASA. Chronic exposure to ecologically relevant concentrations of this drug showed to be mostly ineffective in promoting any significant biochemical alteration in H. diversicolor. However, histochemical observations revealed proliferation of mucous cells in the tegument of chronically exposed individuals to ASA.

Toxicity Assessment of Impacted Sediments from Southeast Coast of Tunisia Using a Biomarker Approach with the Polychaete Hediste diversicolor.

Toxicity caused by exposure to pollutants from marine sediments is a consequence of the interaction between biota and xenobiotics most frequently released by anthropogenic activities. The present work intended to characterize the toxicity of natural sediments putatively impacted by distinct human activities, collected at several sites located in the south of the Gulf of Gabes, Zarzis area, Tunisia. The selected toxicity criteria were analysed following ecologically relevant test conditions. Organisms of the polychaete species Hediste diversicolor were chronically exposed (28 days) to the mentioned sediments. Toxicity endpoints were biomarkers involved in the toxic response to common anthropogenic chemicals, namely neurotoxic (acetylcholinesterase), anti-oxidant (catalase, glutathione peroxidase), metabolic (glutathione S-transferases) enzymatic activities, and oxidative damage (lipid peroxidation, TBARS assay). The chemical characterization of sediments showed that the samples collected from the site near an aquaculture facility were highly contaminated by heavy metals (Cd, Cu, Cr, Hg, Pb, and Zn) and polycyclic aromatic hydrocarbons (fluorene, phenanthrene, anthracene, fluoranthene and pyrene). H. diversicolor individuals exposed to the sediments from this specific site showed the highest values among all tested biomarkers, suggesting that these organisms were possibly under a pro-oxidative stress condition potentially promoted by anthropogenic pollution. Moreover, it was possible to conclude that individuals of the polychaete species H. diversicolor responded to the chronic exposure to potentially contaminated sediments from the southeast coast of Tunisia, eliciting adaptive responses of significant biological meaning.