Drivers of litter ingestion by sea turtles: Three decades of empirical data collected in Atlantic Europe and the Mediterranean.

Sea turtles are considered as bio-indicators for monitoring the efficiency of restoration measures to reduce marine litter impacts on health. However, the lack of extended and standardised empirical data has prevented the accurate analysis of the factors influencing litter ingestion and the relationships with individual health. Historic data collected from 1988 and standard data collected from 2016 were harmonised to enable such analyses on necropsied loggerhead turtles (Caretta caretta) in eight Mediterranean and North-East Atlantic countries. Litter was found in 69.24 % of the 1121 individuals, mostly single-use and fishing-related plastics. Spatial location, sex and life history stage explained a minor part of litter ingestion. While no relationships with health could be detected, indicating that all individuals can be integrated as bio-indicators, the mechanistic models published in literature suggest that the high proportion of plastics in the digestive contents (38.77 % per individual) could have long-term repercussions on population dynamics.

Morphometric Variation among Four Local Ruditapes decussatus Populations in Monastir Bay (Eastern Coast, Tunisia)

Abstract In Tunisia, Monastir bay, extending for 38 km along the eastern coast of Tunisia, harbors different species of bivalves such as the clam Ruditapes decussatus, a species of major socio-economic interest. Despite its importance, no studies have been carried out about the morphology of this mollusc in Monastir bay. Thus, the current study aimed to analyze the shell morphological variations in the European clamRuditapes decussatus from Monastir Bay coast. The comparison of the shell shape of individuals from different populations was performed based on samples taken at four different sites (Khniss, Ksiba, Bkalta and Sokrine). Variance analysis of the morphometric indices (elongation, compactness, convexity and density) for the four populations indicated significant differences among sites with (p=0<0.05). Principal component analysis (PCA) revealed two intra-population groups for Sokrine site, supporting an intra-population variation in the shell morphology. Allometric relationships established between linear variables (shell length, height and width) and ponderal variables (total weight, shell weight) as well as the relative growth between variables (isometry vs. allometry) were analyzed. All morphometric relationships were highly significant (P<0.001) with high correlation coefficients (R2: 0.714-0.982), showing positive allometric growth for Bkalta and negative allometric growth for Sokrine. The data obtained will be useful in research studies and have practical application in diverse research fields such as biology, ecology, fisheries assessment and management.

Data Collection on Marine Litter Ingestion in Sea Turtles and Thresholds for Good Environmental Status.

The following protocol is intended to respond to the requirements set by the European Union's Marine Strategy Framework Directives (MSFD) for the D10C3 Criteria reported in the Commission Decision (EU), related to the amount of litter ingested by marine animals. Standardized methodologies for extracting litter items ingested from dead sea turtles along with guidelines on data analysis are provided. The protocol starts with the collection of dead sea turtles and classification of samples according to the decomposition status. Turtle necropsy must be performed in authorized centers and the protocol described here explains the best procedure for gastrointestinal (GI) tract isolation. The three parts of the GI (esophagus, stomach, intestine) should be separated, opened lengthways and contents filtered using a 1 mm mesh sieve. The article describes the classification and quantification of ingested litter, classifying GI contents into seven different categories of marine litter and two categories of natural remains. The quantity of ingested litter should be reported as total dry mass (weight in grams, with two decimal places) and abundance (number of items). The protocol proposes two possible scenarios to achieve the Good Environmental Status (GES). First: "There should be less than X% of sea turtles having Y g or more plastic in the GI in samples of 50-100 dead turtles from each sub-region", where Y is the average weight of plastic ingested and X% is the percentage of sea turtles with more weight (in grams) of plastic than Y. The second one, which considers the food remain versus plastic as a proxy of individual health, is: "There should be less than X% of sea turtles having more weight of plastic (in grams) than food remains in the GI in samples of 50-100 dead turtles from each sub-region".

Assessing cetacean surveys throughout the Mediterranean Sea: a gap analysis in environmental space.

Heterogeneous data collection in the marine environment has led to large gaps in our knowledge of marine species distributions. To fill these gaps, models calibrated on existing data may be used to predict species distributions in unsampled areas, given that available data are sufficiently representative. Our objective was to evaluate the feasibility of mapping cetacean densities across the entire Mediterranean Sea using models calibrated on available survey data and various environmental covariates. We aggregated 302,481 km of line transect survey effort conducted in the Mediterranean Sea within the past 20 years by many organisations. Survey coverage was highly heterogeneous geographically and seasonally: large data gaps were present in the eastern and southern Mediterranean and in non-summer months. We mapped the extent of interpolation versus extrapolation and the proportion of data nearby in environmental space when models calibrated on existing survey data were used for prediction across the entire Mediterranean Sea. Using model predictions to map cetacean densities in the eastern and southern Mediterranean, characterised by warmer, less productive waters, and more intense eddy activity, would lead to potentially unreliable extrapolations. We stress the need for systematic surveys of cetaceans in these environmentally unique Mediterranean waters, particularly in non-summer months.

Taxonomic observations on stingrays of the genus Dasyatis (Chondrichthyes: Dasyatidae) in the Gulf of Gabès (Southeastern Mediterranean Sea).

Four species of stingrays of the genus Dasyatis occur in the Gulf of Gabès, but the taxonomic status of one of them, D. tortonesei has been questioned by several authors, and synonymized with D. pastinaca by some. The present study, using morphological comparisons and genetic analyses, demonstrates that there is a closely related species to D. pastinaca in the Gulf of Gabès, but distinguishable by several characters. This species is provisionally identified as Dasyatis cf. tortonesei, pending a more extensive comparative study including specimens from other regions of the Mediterranean Sea and eastern North Atlantic Ocean.

Effect of linear alkylbenzene sulfonate (LAS) on human intestinal Caco-2 cells at non cytotoxic concentrations.

Linear alkylbenzene sulfonate (LAS) is a cytotoxic synthetic anionic surfactant widely present in the environment due to its large-scale production and intensive use in the detergency field. In this study, we investigated the effect of LAS (CAS No. 25155-30-0) at non cytotoxic concentrations on human intestinal Caco-2 cells using different in vitro bioassays. As results, LAS increased Caco-2 cell proliferation at concentrations ranging from 1 to 15 ppm, more significantly for shorter exposure time (24 h), confirmed using flow cytometry and trypan blue exclusion methods. Moreover, proteomics analysis revealed that this effect was associated with an over-expression of elongation factor 2 and dipeptidyl peptidase 3, and a down-regulation of 14-3-3 protein theta, confirmed at mRNA level using real-time PCR. These findings suggest that LAS at non cytotoxic concentrations, similar to those observed at wastewater treatment plants outlets, increases the growth rate of colon cancer cells, raising thereby its tumor promotion effect potential.

Treatment of textile wastewater by submerged membrane bioreactor: In vitro bioassays for the assessment of stress response elicited by raw and reclaimed wastewater.

The performance of a pilot-scale membrane bioreactor (MBR) system for the treatment of textile wastewater was investigated. The MBR was continuously operated for 7 months. Very high treatment efficiencies were achieved (color, 100%; chemical oxygen demand (COD), 98%; biochemical oxygen demand (BOD5), 96%; suspended solids (SS), 100%). Furthermore, the MBR treatment efficiency was analyzed from a toxicological-risk assessment point of view, via different In vitro bioassays using Caco-2 cells, a widely used cell model in toxicological studies. Results showed that MBR treatment significantly reduced the raw textile wastewater (RTWW) cytotoxicity on Caco-2 cells by 53% for a hydraulic retention time (HRT) of 2 days. Additionally, the RTWW-induced disruption in the barrier function (BF) of the Caco-2 cell monolayer was also significantly reduced after MBR treatment under a HRT of 2 days (no disruption of BF was observed). Moreover, the effect of RTWW and treated wastewater on stress response was investigated using different stress genes: AHSA1, HSPD1, HSPA1A, HSPA5 and HSPA8. The cell exposure to RTWW significantly increased the expression of all used stress genes; interestingly, the treated wastewater (HRT 2 days) did not show any significant modulation of the stress genes.

Sex ratio estimations of loggerhead sea turtle hatchlings at Kuriat islands, Tunisia: can minor nesting sites contribute to compensate globally female-biased sex ratio?

Hatchling sex ratios in the loggerhead turtle Caretta caretta were estimated by placing electronic temperature recorders in seven nests at Kuriat islands (Tunisia) during the 2013 nesting season. Based on the mean temperatures during the middle third of the incubation period, and on incubation duration, the sex ratio of hatchlings at Kuriat islands was highly male-biased. Presently, the majority of hatchling sex ratio studies are focused on major nesting areas, whereby the sex ratios are universally believed to be heavily female-biased. Here we present findings from a minor nesting site in the Mediterranean, where the hatchling sex ratio was found to be male-biased, suggesting a potential difference between major and minor nesting sites.

Cytotoxic effect of linear alkylbenzene sulfonate on human intestinal Caco-2 cells: associated biomarkers for risk assessment.

Linear alkylbenzene sulfonate (LAS) is a synthetic anionic surfactant widely present in the environment due to its intensive production and use in the detergency field. Admitting that current procedure of risk assessment has limits in providing realistic risk assessment data and predicting the cumulative effect of the toxicant mixtures, the incorporation of information regarding the mode of action and cell response mechanism seems to be a potential solution to overcome these limits. In this regard, we investigated in this study the LAS cytotoxicity on human intestinal Caco-2 cells, trying to unveil the protein actors implicated in the cell response using proteomics approach in order to give a better understanding of the toxicological effect and allow the identification of appropriate biomarkers reflecting the mode of action associated with LAS. As results, we demonstrated that LAS induces a time- and dose-dependent cytotoxicity in Caco-2 cells accompanied by an induction of oxidative stress followed by an excessive increase of intracellular calcium level. Proteomics approach helped in discovering three informative biomarkers of effect associated with LAS cytotoxic effect, reported for the first time: calreticulin, thioredoxin, and heat shock cognate 71 (HSP7C), confirmed by real-time PCR and western blot analysis. These biomarkers could serve for more reliable future risk assessment studies that consider the toxicants mode of action in order to help in the prediction of potential cumulative effects of environmentally coexisting contaminants.

Decreased transfusion needs associated with hydroxyurea therapy in Algerian patients with thalassemia major or intermedia.

BACKGROUND: Studies of evolution of transfusion requirements in thalassemic patients treated with hydroxyurea have produced somewhat conflicting results, especially in patients with thalassemia major. Our aims were to determine the proportion of good responders to hydroxyurea in a population of transfusion-dependent thalassemic patients and to identify the factors associated with a decrease of transfusion needs. STUDY DESIGN AND METHODS: Hydroxyurea treatment was initiated in 9 patients with thalassemia intermedia (TI) and 45 with thalassemia major (TM). Patients received transfusions when their hemoglobin (Hb) levels dropped below 6 g per dL. A decrease in annual transfusion requirements greater than 70 percent defined a good response, between 40 and 70 percent a partial response, and smaller than 40 percent no response. RESULTS: The response was good in 8 (90%) patients with TI and 20 (44.5%) with TM, partial in 9 (20%) patients with TM, and absent in 1 (10%) with TI and 16 (35.5%) with TM. In TM patients, transfusion needs decreased by 56 percent over the first year of hydroxyurea treatment. By univariate analysis, a better response to hydroxyurea was associated with older age at the first transfusion (p = 0.02), higher prehydroxyurea Hb (p = 0.0004), codon 6(-A) mutation (p = 0.002), TI (p = 0.03), and history of splenectomy (p = 0.05). Xmn1-/- was associated with a worse response (p = 0.0001). By multivariate analysis, a better response was associated with the Xmn1 polymorphism (p = 0.008). CONCLUSION: Hydroxyurea may be an alternative to transfusions for TI patients as well as for TM patients in countries that have limited blood supplies.

Hydroxyurea can eliminate transfusion requirements in children with severe beta-thalassemia.

Hydroxyurea (HU) enhances fetal hemoglobin (Hb) production. An increase in total Hb level has been repeatedly reported during HU treatment in patients with sickle cell disease and in several patients with beta-thalassemia intermedia. Effects in patients with beta-thalassemia major are controversial. We now report a marked elevation of total Hb levels with HU that permitted regular transfusions to be stopped in 7 children with transfusion-dependent beta-thalassemia. The median follow-up was 19 +/- 3 months (range, 13-21 months). We conclude that HU can eliminate transfusional needs in children with beta-thalassemia major, which could be particularly useful in countries such as Algeria, where supplies of blood or chelating agents are limited.