Sara Leston

Implications of nutrient decline in the seagrass ecosystem success

The Mondego estuary (Portugal) suffered major changes in environmental quality due to eutrophication, however, in the late 1990 s a restoration project was implemented in order to return the system to its original condition. The main goal of this paper is to evaluate the ecosystem response to the restoration measures applied at three different levels: water quality, primary producers and primary consumers. In post-restoration period a clear decline was observed in dissolved inorganic nitrogen which was reflected in the gradual recovery of Zostera noltii and a concomitant decline in green macroalgae. Macrobenthic assemblages responded variably to the recovery process. In the seagrass bed and intermediate area, there was a large increase in total biomass, but in the eutrophic area species diversity increased. Despite improvement in the ecological status of the system, full recovery has not been achieved yet, possible due to hysteresis in the dynamics of this system.

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 CCβ 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. Graphical Abstract

Distribution of Corbicula fluminea (Müller, 1774) in the invaded range: a geographic approach with notes on species traits variability

Corbicula fluminea is considered one of the most important non-native invasive species (NIS) in aquatic systems mainly due to its widespread distribution and ecological and economic impacts. This species is known to negatively affect native bivalves, also with severe effects on biodiversity and ecosystem functioning. Throughout an exhaustive bibliographic survey and with the aid of Geographic Information Systems tools, this study tracks the species dispersion from its native range, including the description of important physical and environmental barriers. Additional analyses were conducted to examine possible influences of latitudinal/temperature gradients on important traits (e.g. life span, maximum and mean body length, growth at the end of first year). Altitude and winter minimum temperature appear to be delaying the invasion worldwide, but it seems inevitable that the species will spread across the globe. Latitude and summer temperature show a relationship with growth and life span. Overall, the information gathered in this review may be relevant to forecast future distribution patterns of this NIS, and to anticipate the possible implementation of effective management measures. Moreover, it may constitute a valuable tool in the prediction of population responses to an increasingly changing environment.

Hepatic glycogen synthesis in farmed European seabass (Dicentrarchus labrax L.) is dominated by indirect pathway fluxes

Hepatic glycogen synthesis fluxes from direct and indirect pathways were quantified in seabass by postmortem (2)H NMR analysis of plasma water (PW) and glycogen glucosyl (2)H enrichments from (2)H-enriched seawater. Eighteen fish (28.0 ± 1.7 cm and 218.0 ± 43.0 g) were divided into three groups of 6 and studied over 24 days with transfer to 5% (2)H-seawater after day 21. Over this period, one group was fed daily with fishmeal, a second group was fasted, and a third group was fasted for 21 days followed by 3 days refeeding. Glycogen turnover and sources were determined from the ratio of glucosyl position 5 enrichment to that of plasma water (H5/PW). Glycogen levels of fed fish were significantly higher than fasted (665.4 ± 345.2 μmol.g(-1) liver versus 77.2 ± 59.5 μmol.g(-1) liver, P<0.05) while refed fish had comparable levels to fed (584.6 ± 140.4 μmol.g(-1) liver). Glycogen enrichment of fed fish was undetectable indicating negligible turnover over 3 days. For fasted fish, H5/PW was ~50% indicating that half of the glycogen had turned over via indirect pathway flux. For refed fish, H5/PW was ~100% indicating that the indirect pathway accounted for all net glycogen synthesis. Direct pathway conversion of dietary carbohydrate to glycogen was not detected in any of the groups.

Analysis of glucose metabolism in farmed European sea bass (Dicentrarchus labrax L.) using deuterated water

Glucose metabolism in free-swimming fasted and fed seabass was studied using deuterated water ((2)H(2)O). After transfer to seawater enriched with 4.9% (2)H(2)O for 6-h or for 72-h, positional and mole percent enrichment (MPE) of plasma glucose and water were quantified by (2)H NMR and ESI-MS/MS. Plasma water (2)H-enrichment reached that of seawater within 6h. In both fasted and fed fish, plasma glucose MPE increased asymptotically attaining ~55% of plasma water enrichment by 72 h. The distribution of (2)H-enrichment between the different glucose positions was relatively uniform. The gluconeogenic contribution to glucose that was synthesized during (2)H(2)O administration was estimated from the ratio of position 5 and 2 glucose enrichments. For both fed and fasted fish, gluconeogenesis accounted for 98±1% of the glucose that was produced during the 72-h (2)H(2)O administration period. For fasted fish, gluconeogenic contributions measured after 6h were identical to 72-h values (94±3%). For fed fish, the apparent gluconeogenic contribution at 6-h was significantly lower compared to 72-h (79±5% versus 98±1%, p<0.05). This may reflect a brief augmentation of gluconeogenic flux by glycogenolysis after feeding and/or selective enrichment of plasma glucose position 2 via futile glucose-glucose-6-phosphate cycling.

The influence of sulfathiazole on the macroalgae Ulva lactuca

Sulfonamides (SA) are a class of antibiotics routinely found in environmental matrices and therefore their role as contaminants should be investigated in non-target organisms. With this purpose the present experimental work has evaluated the exposure of the chlorophycean Ulva lactuca L. to sulfathiazole (STZ), a SA drug commonly used in aquaculture, at two concentrations representing prophylactic (25 μg mL(-1)) and therapeutic (50 μg mL(-1)) administrations. Results showed that STZ exhibits high stability in seawater with only 18% degradation over the 5d assay at both dosages tested. Also, macroalgae demonstrated an efficient uptake capacity with constant internal concentrations after 24h regardless of the external solutions and thus should be considered as a bioindicator species in risk assessment. Both STZ concentrations induced a slight inhibition of the macroalgae growth after 96 h.

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.

The effects of the nitrofuran furaltadone on Ulva lactuca

The use of pharmaceuticals in the food production industry as prophylatic and therapeutic agents is necessary to promote animal health, but may entail significant consequences to natural ecosystems, especially in the cases of overdosing and use of banned pharmaceuticals. The vast effects that antibiotics released into the environment have on non-target organisms are already under the scope of researchers but little attention has been given to primary producers such as macroalgae. The present study assessed furaltadones, an antibacterial agent illegally used for veterinary purposes, uptake capacity by Ulva lactuca and its effect in the growth of this cosmopolitan macroalgae. Differences in macroalgal growth were shown when submitted to prophylactic and therapeutic concentrations of furaltadone in the water (16 and 32 μg mL⁻¹, respectively). The therapeutic concentration caused higher growth impairment than the prophylactic treatment did, with 87.5% and 58% reductions respectively. Furthermore, together with data collected from the accumulation assays, with values of internal concentrations as high as 18.84 μg g⁻¹ WW, suggest that the macroalgae U. lactuca should be included in field surveys as a biomonitor for the detection of nitrofurans.

The effects of chloramphenicol on Ulva lactuca

The administration of pharmacological substances in the food producing industry is a crucial and long established practice in ensuring animal welfare. However, a very high percentage of the drugs used will directly or indirectly be present in the various compartments of natural ecosystems therefore constituting a source of pollution. The reactions that these active compounds may impose on non-target organisms are still widely unknown and further research is essential. Also, new approaches on monitoring are necessary and in this sense, the present work aimed to assess the persistence of chloramphenicol (a banned but illegally used antibiotic) in seawater, together with its effects on the growth of the green macroalgae Ulva lactuca. Moreover, the potential use of this species as a bioindicator was assessed. Results showed CAP presented an exponential degradation pattern in seawater with concentrations decreasing faster than expected. As for the effects on U. lactuca it acted as a growth promoter also contradicting the initial assumptions. Regarding the role of this species in biomonitoring it successfully took up CAP in solution while reflecting the concentrations present conferring it good characteristics as a bioindicator. On the other hand, this ability points to a possibility of CAP being accumulated and transferred along the trophic web through the consumption of U. lactuca by organisms in higher levels.

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.