Soil pH influences the toxicity of Basamid® eluates to non-target species of primary consumers.

Basamid® is a fumigant nematicide and fungicide known to break down in several volatile compounds, mainly methyl isothiocyanate (MITC), when in contact with water. Soil abiotic parameters, such as pH, influences this breakdown process, and thus, the toxic effects of Basamid® to aquatic biota. This work studied the influence of soil pH (5.5, 6.5 and 7.5) on the toxicity of eluates (1:4, m:v), obtained from Basamid®-contaminated soils (with the recommended dose of 145 mg of dazomet/Kg of soil), on two primary consumers: Daphnia magna and Brachionus calyciflorus. For this, lethal and sublethal toxicity of eluates originated from soils at pH 5.5, 6.5 and 7.5, contaminated with Basamid® (Ba-E 5.5; 6.5 and 7.5, respectively), were assessed (dilutions between 0.096 - 100%). The LD50,24h of Basamid® eluates for D. magna varied from 3.07% to 7.82% (Ba-E 6.5 and Ba-E 5.5 respectively), while for B. calyciflorus varied from 18.1% to 84.7% (Ba-E 6.5 and Ba-E 7.5, respectively). Both species were less sensitive to Basamid® eluates originated from soils with pH 7.5 and more sensitive to those obtained from soils with pH 6.5. Regarding the sublethal effects, a lower soil pH was associated with a higher toxicity of Basamid® to D. magna reproduction (LOED: 0.125% Ba-E 5.5), while for B. calyciflorus such a higher toxicity was observed at the highest soil pH (ED20: 7.42% [5.10-9.74] at Ba-E 7.5). These results show a negative association between soil pH and the lethal toxicity of Basamid® contaminated eluates. However, such a pattern was not observed at sublethal level, at which a species dependency was observed regarding the influence of soil pH in the observed toxicity. Nevertheless, it is to highlight that very low concentrations of eluates (as 3.07%) caused significant mortality, indicating a high risk for freshwater biota. Considering that Basamid® is likely to reach the aquatic systems is real, for which reason the recommended dose must be reviewed at environmentally-relevant scenarios.

Assessing pharmaceuticals in the green seaweed Ulva lactuca through a multi-residue UHPLC-ToF-MS strategy.

Seaweeds have become an important asset in several sectors, including the food and feed industries, cosmetics, and pharmaceuticals, among others. Whether harvested or reared, interest in algae has been growing worldwide due to the resources they offer, including proteins, vitamins, minerals, carbohydrates, essential fatty acids, and dietary fiber, as well as sources of biologically active compounds. However, given their morphology and physiology, as well as their harvest and cultivation environments, algae are prone to the presence of hazards, including pharmaceuticals taken up from the water. Thus, to ensure human and animal safety as well as environmental health, monitoring is essential. Therefore, this work describes the development and validation of a sensitive screening and confirmatory analytical method based on ultra-high-performance liquid chromatography coupled with time-of-flight mass spectrometry (UHPLC-ToF-MS). This multi-residue method enables the determination of 62 pharmaceuticals distributed between 8 therapeutic classes and was fully validated according to Commission Implementing Regulation (EU) 2021/808.

Ecotoxicity of eluates obtained from Basamid® contaminated soils is pH dependent: A study with Hydra viridissima, Xenopus laevis and Danio rerio.

Agrochemicals are mostly used to deplete pests and treat diseases in terrestrial agro-ecosystems. However, their transport through the soil, by leaching and/or runoff, may cause them to reach aquatic systems. Environmental parameters, such as soil pH, can affect this transport, by influencing the magnitude of agrochemicals degradation and chemical reaction. This work aimed at investigating the influence of soil pH on the toxicity of eluates obtained from Basamid® contaminated soils to Hydra viridissima, Xenopus laevis and Danio rerio. For this, a natural soil with pH amended to 5.5, 6.5 and 7.5, was spiked with the recommended dose (RD) of Basamid® (145 mg dazomet/kg soil) and eluates (Ba-E) were prepared with the respective species culture medium. Dilutions of the eluates (0.14-100%), obtained from the three soils (Ba-E 5.5, Ba-E 6.5 and Ba-E 7.5, corresponding to soil spiked with Basamid® RD at soil pH of 5.5, 6.5 and 7.5, respectively), were used to expose the organisms. Results showed that for H. viridissima increased soil alkalinity provoked less mortality comparatively to lower soil pH [LD50,96h of Ba-E 5.5: 10.6% and LD50,96h of Ba-E 7.5: 21.2%]. As for X. laevis and D. rerio Ba-E lethal ecotoxicity was similar across soil pH (LD50,96h varied from 5.7 to 6.9% and from 2.1 to 4.3%, respectively). For malformations, 20% effect dilution (ED) in H. viridissima was significantly higher at Ba-E 7.5 (ED20,96h: 17.4%), comparatively to Ba-E 5.5 and Ba-E 6.5 (ED20,96h: 7.9% and 7.7%, respectively). From the three tested organisms and based on both lethal and sublethal effects, H. viridissima presented the highest tolerance to Basamid® eluates and soil pH was a major factor determining the fumigant toxicity, with higher soil pH levels inducing, lower toxicity. The eluates obtained from soils contaminated with RD of Basamid® induced severe effects to the three aquatic species.

Soil pH matters in the ecotoxicity of Basamid® to freshwater microalgae and macrophytes.

Intensive agriculture along with the use of agrochemicals has been associated with low soil fertility, soil erosion, and soil acidity. Management of soil pH through liming is a common practice in agriculture to increase soil fertility and nutrient availability. When altering soil pH, different chemical reactions occur depending on soil composition and agrochemicals presence. Basamid® is a fumigant used worldwide targeting soil nematodes, fungi, and weeds in diverse crops, that can reach freshwater ecosystems by leaching through the soil layers. The major goal of this work was to assess the influence of soil pH in the toxicity of Basamid® eluates to the microalgae Raphidocelis subcapitata and the duckweed Lemna minor. For this, eluates were prepared from soils with different pH (5.5, 6.5 and 7.5), contaminated with the recommended dose of Basamid® corresponding to 145.7 mg of dazomet/Kg soil. Soil was amended with calcium carbonate (CaCO3). Raphidocelis subcapitata and L. minor were exposed to the eluates during 72 h and 7 days respectively, and multiple endpoints were assessed: growth rate, biomass, pigment as chlorophyl content and cell damage. Results showed that soil pH can influence the performance of the tested species and also be a major factor in influencing Basamid®'s toxicity. However, a clear pattern of the influence of soil pH on Basamid®'s toxicity was not observed and was species dependent. For R. subcapitata lower soil pHs induced higher toxicity of Basamid®'s to the algae [ED50 for growth rate: 30 % (confidence limits-CL: 22.8-37.2) for soil pH 5.5; >100 % for soil pH 6.5 and pH 7.5], while for L. minor the opposite was observed [ED50 for number of fronds: 27.2 % (CL: 22.8-31.6) for pH 5.5; 20.3 % (CL: 10.0-30.6) for pH 6.5 and 10.7 % (CL: 6.3-15.1)]. Overall, these results showed that leachates of Basamid® through soils, at recommended doses, can have a severe impact on aquatic systems, with or without the influence of abiotic factors.

Does an Invasive Bivalve Outperform Its Native Congener in a Heat Wave Scenario? A Laboratory Study Case with Ruditapes decussatus and R. philippinarum.

Global warming and the subsequent increase in the frequency of temperature anomalies are expected to affect marine and estuarine species' population dynamics, latitudinal distribution, and fitness, allowing non-native opportunistic species to invade and thrive in new geographical areas. Bivalves represent a significant percentage of the benthic biomass in marine ecosystems worldwide, often with commercial interest, while mediating fundamental ecological processes. To understand how these temperature anomalies contribute to the success (or not) of biological invasions, two closely related species, the native Ruditapes decussatus and the introduced R. philippinarum, were exposed to a simulated heat wave. Organisms of both species were exposed to mean summer temperature (~18 °C) for 6 days, followed by 6 days of simulated heat wave conditions (~22 °C). Both species were analysed for key ecological processes such as bioturbation and nutrient generation-which are significant proxies for benthic function and habitat quality-and subcellular biomarkers-oxidative stress and damage, and energetic metabolism. Results showed subcellular responses to heat waves. However, such responses were not expressed at the addressed ecological levels. The subcellular responses to the heat wave in the invasive R. philippinarum pinpoint less damage and higher cellular energy allocation to cope with thermal stress, which may further improve its fitness and thus invasiveness behaviour.

Assessment of fipronil toxicity to the freshwater midge Chironomus riparius: Molecular, biochemical, and organismal responses.

Fipronil is a phenylpyrazole insecticide that entered the market to replace organochlorides and organophosphates. Fipronil impairs the regular inhibition of nerve impulses that ultimately result in paralysis and death of insects. Because of its use as a pest control, and due to runoff events, fipronil has been detected in freshwater systems near agricultural areas, and therefore might represent a threat to non-target aquatic organisms. In this study, the toxicity of fipronil to the freshwater midge Chironomus riparius was investigated at biochemical, molecular, and whole organism (e.g. growth, emergence, and behavior) levels. At the individual level, chronic (28 days) exposure to fipronil resulted in reduced larval growth and emergence with a lowest observed effect concentration (LOEC) of 0.081 µg L-1. Adult weight, which is directly linked to the flying performance and fecundity of midges, was also affected (LOEC = 0.040 µg L-1). Additionally, behavioral changes such as irregular burrowing behavior of C. riparius larvae (EC50 = 0.084 µg L-1) and impairment of adult flying performance were observed. At a biochemical level, acute (48 h) exposure to fipronil increased cellular oxygen consumption (as indicated by the increase of electron transport system (ETS) activity) and decreased antioxidant and detoxification defenses (as suggested by the decrease in catalase (CAT) and glutathione S-transferase (GST) activities). Exposure to fipronil also caused alterations in the fatty acid profile of C. riparius, since high levels of stearidonic acid (SDA) were observed. A comparison between exposed and non-exposed larvae also revealed alterations in the expression of globins, cytoskeleton and motor proteins, and proteins involved in protein biosynthesis. These alterations may aid in the interpretation of potential mechanisms of action that lead to the effects observed at the organism level. Present results show that environmentally relevant concentrations of fipronil are toxic to chironomid populations which call for monitoring of phenylpyrazole insecticides and of their ecological effects in freshwaters. Present results also emphasize the importance of complementing ecotoxicological data with molecular approaches such as proteomics, for a better interpretation of the mode of action of insecticides in aquatic invertebrates.

Fate and effects of two pesticide formulations in the invertebrate Folsomia candida using a natural agricultural soil.

Degradation rates of two widely used pesticides were assessed, and acute and chronic effects on a standard invertebrate species investigated. An herbicide (Montana®) and fungicide (Bravo500®) formulations were investigated and results were compared to the isolated active substances of each formulation (glyphosate and chlorothalonil, respectively). Tests were performed using the invertebrate Folsomia candida as test species and an agricultural natural soil. Degradation rate tests were determined under aerobic conditions at 20 ±â€¯2 °C, using an ecologically relevant concentration of 5 mg (a.i.) kg-1 of soil for both chemicals. Results demonstrated degradation half-lives (DT50) of 2.2 days for Montana® and 2.8 days when pure glyphosate was tested. Values of 1.1 and 2.9 days were registered for Bravo500® and its active substance chlorothalonil, respectively. There were no effects on survival for the tested concentrations of both forms of the herbicide (up to 17.3 mg kg-1). However, reproduction was affected, but only by the herbicide formulation, with an estimated EC50 value of 4.63 mg (a.i.) kg-1. Effects were most unlikely related to glyphosate. For chlorothalonil, both tested forms affected survival and reproduction. The estimated LC50 values were 117 mg (a.i.) kg-1 and 73.5 mg (a.i.) kg-1, and the EC50 41.3 mg (a.i.) kg-1 and 14.9 mg kg-1 for the formulation and the active ingredient, respectively. The effects of the active ingredient were significantly stronger, indicating the major influence of the active substance in the effects caused also by the formulation. Overall results demonstrate the importance of evaluating the effects of the formulated chemicals, as they are applied in the field, and not only their isolated active ingredients.

From laboratory to the field: Validating molecular markers of effect in Folsomia candida exposed to a fungicide-based formulation.

Under controlled laboratory conditions, toxicity data tend to be less variable than in more realistic in-field studies and responses may thus differ from those in the natural environment, creating uncertainty. The validation of data under environmental conditions is therefore a major asset in environmental risk assessment of chemicals. The present study aimed to validate the mode of action of a commercial fungicide formulation in the soil invertebrate F. candida, under more realistic exposure scenarios (in-field bioassay), by targeting specific molecular biomarkers retrieved from laboratory experiments. Organisms were exposed in soil cores under minimally controlled field conditions for 4 days to a chlorothalonil fungicide dosage causing 75% reduction of reproduction in a previous laboratory experiment (127 mg a.i. kg-1) and half this concentration (60 mg a.i. kg-1). After exposure, organisms were retrieved and RNA was extracted from each pool of organisms. According to previous laboratorial omics results with the same formulation, ten genes were selected for gene expression analysis by qRT-PCR, corresponding to key genes of affected biological pathways including glutathione metabolism, oxidation-reduction, body morphogenesis, and reproduction. Six of these genes presented a dose-response trend with higher up- or down-regulation with increasing pesticide concentrations. Highly significant correlations between their expression patterns in laboratory and in-field experiments were observed. This work shows that effects of toxicants can be clearly demonstrated in more realistic conditions using validated biomarkers. Our work outlines a set of genes that can be used to assess the early effects of pesticides in a realistic agricultural scenario.

Development and validation of a multi-residue and multi-class screening method of 44 antibiotics in salmon (Salmo salar) using ultra-high-performance liquid chromatography/time-of-flight mass spectrometry: Application to farmed salmon.

A fast and sensitive multi-residue and multiclass screening method for the simultaneous determination of 44 antimicrobials in salmon muscle, using ultra- high-performance liquid chromatography/time-of-flight mass spectrometry (UHPLC-TOF/MS), was develop and validated. Two different procedures for the extraction step were tested, and an extraction with acetonitrile, ethylenediaminetetra acetic acid (EDTA) and n-hexane proved to be the best alternative. The method was validated, in accordance with Decision 2002/657/EC, using a qualitative approach at the CCß level. The detection of the analytes was accomplished by retention time and accurate mass, whose maximum error should not exceed 5 ppm. All the compounds were successfully detected and identified at concentration levels corresponding to ½ maximum residue limit (MRL). The screening method was applied to 39 store bought samples of farmed salmon purchased in Portugal, originating from Norway and Denmark, and no antibiotic residues were detected.

Detection and quantification of 47 antibiotic residues in farmed European sea bass (Dicentrarchus labrax) using a multi-class and multi-residue UHPLC-MS/MS method.

In the present study, a multi-class multi-residue method for the simultaneous detection and determination of antibiotics in European sea bass (Dicentrarchus labrax) was developed and validated. The method based on UHPLC-MS/MS proved to be a rapid, highly selective and sensitive tool, requiring minimum sample preparation, for screening and detection of 47 compounds from eight different classes. The validation was performed according to EU regulation 2002/657/EC, proving the method's suitability for application in routine analysis. The method was applied to the analysis of 30 samples of farmed European sea bass purchased in different supermarkets in Portugal. Antibacterial residues were detected in 6 of the 30 analysed samples, namely enrofloxacin and oxytetracycline, in concentrations ranging from 0.1 to 12 µg kg-1.

Oxytetracycline accumulation in the macroalgae Ulva: Potential risks for IMTA systems.

Oxytetracycline (OTC) is one of the most used antibiotics in aquaculture. With the development of Integrated Multitrophic Aquaculture (IMTA) systems in order to mitigate some aquacultures' adverse effects, attention needs to be shifted to other co-cultured species that can also accumulate such pharmaceuticals and pose a risk to human consumption. Therefore, the present work evaluated the exposure of the seaweed Ulva to OTC at two realistic concentrations (0.040 and 0.120 mg L-1). Oxytetracycline degradation rates in seawater were dependent on the initial concentration but were not influenced by the presence of Ulva. The macroalgae presented good assimilation rates of OTC, with internal concentrations reaching 40.9934 ng g-1 WW for the lowest concentration tested and 108.6787 ng g-1 WW for the highest, with a steep decrease after 48 and 24 h, respectively. Nonetheless, concentrations were still half of the Maximum Residue Limit set for fish (100 µg kg-1) 48 h after C2 treatment. The highest dosage tested stimulated growth 96 h after the beginning of the trial, although some signs of decay could also be found in Ulva's fronds.

Screening of human and veterinary pharmaceuticals in estuarine waters: A baseline assessment for the Tejo estuary.

We investigated the presence of 66 human and veterinary pharmaceuticals from seven therapeutic groups in surface waters of the Tejo estuary. Collection sites covered the entire estuary and included areas near main river inflows and wastewater treatment outfalls, traversing urban, agriculture, aquaculture, and nature reserve areas. Detection of pharmaceuticals was performed via UHPLC-TOF-MS. Pharmaceuticals were found in all sites (32 different compounds in total). Antibiotics, ß-blockers, antihypertensives and anti-inflammatories were the most frequently detected (>90%), with variation in concentrations reflecting the multifaceted nature of estuarine surroundings (accumulated site contamination between 15 and 351 ng L-1). Higher concentrations of antidepressant Sertraline (304 ng L-1), non-steroidal anti-inflammatory Diclofenac (51.8 ng L-1), lipid regulator Gemfibrozil (77.0 ng L-1), antihypertensive Ibersartan (161.9 ng L-1) or antibiotic Doxycycline (128.0 ng L-1), among others, though localized may potentially impact key estuarine functions or services. Ultimately, results provide a baseline for regulatory information and future biota evaluations.

Ecological consequences of invasion across the freshwater-marine transition in a warming world.

The freshwater-marine transition that characterizes an estuarine system can provide multiple entry options for invading species, yet the relative importance of this gradient in determining the functional contribution of invading species has received little attention. The ecological consequences of species invasion are routinely evaluated within a freshwater versus marine context, even though many invasive species can inhabit a wide range of salinities. We investigate the functional consequences of different sizes of Corbicula fluminea-an invasive species able to adapt to a wide range of temperatures and salinity-across the freshwater-marine transition in the presence versus absence of warming. Specifically, we characterize how C. fluminea affect fluid and particle transport, important processes in mediating nutrient cycling (NH 4-N, NO 3-N, PO 4-P). Results showed that sediment particle reworking (bioturbation) tends to be influenced by size and to a lesser extent, temperature and salinity; nutrient concentrations are influenced by different interactions between all variables (salinity, temperature, and size class). Our findings demonstrate the highly context-dependent nature of the ecosystem consequences of invasion and highlight the potential for species to simultaneously occupy multiple components of an ecosystem. Recognizing of this aspect of invasibility is fundamental to management and conservation efforts, particularly as freshwater and marine systems tend to be compartmentalized rather than be treated as a contiguous unit. We conclude that more comprehensive appreciation of the distribution of invasive species across adjacent habitats and different seasons is urgently needed to allow the true extent of biological introductions, and their ecological consequences, to be fully realized.

Effects of the neonicotinoids acetamiprid and thiacloprid in their commercial formulations on soil fauna.

Neonicotinoids are the most prominent group of insecticides in the world and are commercialized in over 120 countries for the control of agricultural pests mainly due to their broad-spectrum activity and versatility in application. Though non-target soil organisms are likely to be exposed during application, there is paucity of information in scientific literature regarding their sensitivity to neonicotinoids. This study attempts to fill this gap by evaluating, under laboratory conditions, the chronic toxicity of the neonicotinoids thiacloprid and acetamiprid, through their commercial formulations (CF), to the soil invertebrates Folsomia candida, Eisenia andrei and Enchytraeus crypticus. Results obtained indicate that the relative reproductive sensitivity of the test organisms can be expressed as: F. candida = E. andrei > E. crypticus (for acetamiprid CF) and E. andrei > F. candida > E. crypticus (for thiacloprid CF). To extrapolate from laboratory test results to field conditions, predicted environmental concentrations (PECs) and predicted no-effect concentrations were derived. Calculated toxicity-exposure ratios (TER = EC10/PEC) were below trigger values for acetamiprid and thiacloprid, when estimated with initial PEC. While estimated hazard quotients (HQ = PEC/PNEC), were greater than the European Commission trigger value. Therefore, with the current data under standard environmental risk assessment schemes it can be considered that the risk of thiacloprid and acetamiprid to the soil compartment is unacceptable. However, further research into the effects of these substances on different organisms is required to increase the confidence in the risk assessment estimates for instance, by calculating hazardous concentrations using species sensitivity distribution curves.

A multiresidue approach for the simultaneous quantification of antibiotics in macroalgae by ultra-high performance liquid chromatography-tandem mass spectrometry.

Together with fish, algae reared in aquaculture systems have gained importance in the last years, for many purposes. Besides their use as biofilters of effluents, macroalgae's rich nutritional profiles have increased their inclusion in human diets but also in animal feeds as sources of fatty acids, especially important for the fish industry. Nonetheless, algae are continuously exposed to environmental contaminants including antibiotics and possess the ability for bioaccumulation of such compounds. Therefore, the present paper describes the development and validation of an ultra-high performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous quantification of antibiotics in the green macroalgae Ulva lactuca. This multi-residue method enables the determination of 38 compounds distributed between seven classes and was fully validated according to EU Decision 2002/657/EC.

Ecotoxicological effects of pig manure on Folsomia candida in subtropical Brazilian soils.

The effects of pig manure, from diets incorporating veterinary pharmaceuticals, on survival and reproduction of Folsomia candida were evaluated. Manures derived from the following diets: corn and soymeal (CS); 85% CS diet+15% wheat meal (TR); CS diet+100ppm doxycycline+50ppm colistin+2500ppm Zn oxide (CSa); TR diet+100ppm doxycycline+50ppm colistin+2500ppm Zn oxide (TRa). Manures were tested in two subtropical soils representative of southern (Oxisol and Entisol). Despite the antibiotics no significant differences were found between the four manures within each soil. However, strong differences were found on the toxicity between soils. In Oxisol, LC50 values were around 100m(3)ha(-1), and EC50 values around 80m(3)ha(-1). In Entisol these were much lower, with LC50 values oscillating around 20m(3)ha(-1) and EC50 values between 10-15m(3)ha(-1). The observed toxicity on both soils was attributed to excess of nitrogen, Cu and Zn in the highest doses. The strong difference between soils could be explained by soil properties, namely CEC, organic matter, and clay contents that were lower in Entisol, indicating a poor ability to retain contaminants increasing their availability in soil. Results suggest that the application of these residues should be regulated not only using a volume-based criterion, but should incorporate data on soil properties, complemented by an ecotoxicological assessment.

Contribution of dietary starch to hepatic and systemic carbohydrate fluxes in European seabass (Dicentrarchus labrax L.).

In the present study, the effects of partial substitution of dietary protein by digestible starch on endogenous glucose production were evaluated in European seabass (Dicentrarchus labrax). The fractional contribution of dietary carbohydrates v. gluconeogenesis to blood glucose appearance and hepatic glycogen synthesis was quantified in two groups of seabass fed with a diet containing 30% digestible starch (DS) or without a carbohydrate supplement as the control (CTRL). Measurements were performed by transferring the fish to a tank containing water enriched with 5% (2)H2O over the last six feeding days, and quantifying the incorporation of (2)H into blood glucose and hepatic glycogen by (2)H NMR. For CTRL fish, gluconeogenesis accounted for the majority of circulating glucose while for the DS fish, this contribution was significantly lower (CTRL 85 (SEM 4) % v. DS 54 (SEM 2) %; P < 0.001). Hepatic glycogen synthesis via gluconeogenesis (indirect pathway) was also significantly reduced in the DS fish, in both relative (CTRL 100 (SEM 1) % v. DS 72 (SEM 1) %; P < 0.001) and absolute terms (CTRL 28 (SEM 1) v. DS 17 (sem 1) µmol/kg per h; P < 0.001). A major fraction of the dietary carbohydrates that contributed to blood glucose appearance (33 (sem 1) % of the total 47 (SEM 2) %) had undergone exchange with hepatic glucose 6-phosphate. This indicated the simultaneous activity of hepatic glucokinase and glucose 6-phosphatase. In conclusion, supplementation of digestible starch resulted in a significant reduction of gluconeogenic contributions to systemic glucose appearance and hepatic glycogen synthesis.

Analysis of chloramphenicol residues in the macroalgae Ulva lactuca through ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS).

Antibiotic use is a well-described practice to promote animal health whether for prevention or treatment. Nonetheless, it can also cause a number of potentially harmful effects that dictate the need to implement regulation to assure a reduction of hazards to the consumers and the environment. Chloramphenicol (CAP) is a broad-spectrum antibacterial excluded from use in animal food production but despite this, reports of illegal use still persist. More recently, awareness has risen that the surrounding natural ecosystems can potentially be contaminated by pharmaceuticals and the extent of their effects in non-target organisms is already under the scope of researchers. To face the demanding new challenges a methodology for the determination of CAP in the green macroalgae Ulva lactuca by ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) was developed, optimized and fully validated following the guidelines of the EC Decision 2002/657.

Sulfathiazole: analytical methods for quantification in seawater and macroalgae.

The awareness of the interconnection between pharmaceutical residues, human health, and aquaculture has highlighted the concern with the potential harmful effects it can induce. Furthermore, to better understand the consequences more research is needed and to achieve that new methodologies on the detection and quantification of pharmaceuticals are necessary. Antibiotics are a major class of drugs included in the designation of emerging contaminants, representing a high risk to natural ecosystems. Among the most prescribed are sulfonamides, with sulfathiazole being the selected compound to be investigated in this study. In the environment, macroalgae are an important group of producers, continuously exposed to contaminants, with a significant role in the trophic web. Due to these characteristics are already under scope for the possibility of being used as bioindicators. The present study describes two new methodologies based on liquid chromatography for the determination of sulfathiazole in seawater and in the green macroalgae Ulva lactuca. Results show both methods were validated according to international standards, with MS/MS detection showing more sensitivity as expected with LODs of 2.79ng/g and 1.40ng/mL for algae and seawater, respectively. As for UV detection the values presented were respectively 2.83µg/g and 2.88µg/mL, making it more suitable for samples originated in more contaminated sites. The methods were also applied to experimental data with success with results showing macroalgae have potential use as indicators of contamination.

Multi-residue and multi-class determination of antibiotics in gilthead sea bream (Sparus aurata) by ultra high-performance liquid chromatography-tandem mass spectrometry.

This paper describes a method for the determination of 41 antibiotics from seven different classes in gilthead sea bream (Sparus aurata) by ultra-high-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS). Sulfonamides, trimethoprim, tetracyclines, macrolides, quinolones, penicillins and chloramphenicol were simultaneously determined. Fourteen procedures for sample treatment were tested and an extraction with acetonitrile and ethylenediaminetetra acetic acid was found to be the best option. The methodology was validated in accordance with Decision 2002/657/EC. Precision in terms of relative standard deviation (RSD) was under 17% for all compounds, and the recoveries ranged from 92% to 111%. CCα and CCß were determined according to the maximum residue limit or the minimum required performance limit, when necessary. The validation provided evidence that the method was suitable for application in routine analysis for the detection and confirmation of antibiotics in muscle of gilthead sea bream, an important and intensively produced fish in aquaculture.