Contamination of planktonic food webs in the Mediterranean Sea: Setting the frame for the MERITE-HIPPOCAMPE oceanographic cruise (spring 2019).

This paper looks at experiential feedback and the technical and scientific challenges tied to the MERITE-HIPPOCAMPE cruise that took place in the Mediterranean Sea in spring 2019. This cruise proposes an innovative approach to investigate the accumulation and transfer of inorganic and organic contaminants within the planktonic food webs. We present detailed information on how the cruise worked, including 1) the cruise track and sampling stations, 2) the overall strategy, based mainly on the collection of plankton, suspended particles and water at the deep chlorophyll maximum, and the separation of these particles and planktonic organisms into various size fractions, as well as the collection of atmospheric deposition, 3) the operations performed and material used at each station, and 4) the sequence of operations and main parameters analysed. The paper also provides the main environmental conditions that were prevailing during the campaign. Lastly, we present the types of articles produced based on work completed by the cruise that are part of this special issue.

Karenia selliformis bloom dynamics and growth rate estimation in the Sfax harbour (Tunisia), by using automated flow cytometry equipped with image in flow, during autumn 2019.

A Karenia selliformis bloom event in the Gulf of Gabès (Mediterranean Sea), was monitored over 9 days at high frequency during fall 2019, by using an automated flow cytometer (Cytosense, Cytobuoy b.v.) with an image-in-flow attachment. The instrument recorded the shape of the optical signals that lead to the resolution of six cell groups of pico-, nano- and microphytoplankton, during the Harmful Algal Bloom (HAB). K. selliformis cell dimensions derived from the hourly records, enabled to estimate the daily division rate over the bloom period. Results revealed that K. selliformis was the only bloom-forming species and it reached its highest mean abundance the fourth day of the survey. A shift in the nutrient composition occurred with a potential P limitation during the bloom growth and N limitation during the bloom collapse. The co-inertia analysis revealed opposite patterns for K. selliformis and heterotrophic prokaryotes suggesting trophic interactions and possible mixotrophic behaviour of K. selliformis at the end of the bloom. K. selliformis exhibited low growth rates generally < 1 division day-1, which could not explain the observed high abundance. The tide played a crucial role in the dynamics of K. selliformis at a semi-diurnal scale and at spring-neap tide scale and was probably enhancing K. selliformis accumulation.

Essential oil composition and biological activities of Ononis alba Poir (Fabaceae).

The phytochemical and biological properties of Ononis alba Poir L. (Fabaceae) were investigated for the first time in this study. The chemical composition of the essential oil obtained from the aerial parts was analysed by GC-MS. The phenolic contents of extracts obtained with different solvents were determined by the Folin-Ciocalteu assay and the antioxidant activity was evaluated through DPPH and CUPRAC methods. The inhibitory potential of these extracts was evaluated on α-amylase and α-glucosidase, whereas the antimicrobial effect was verified against some bacteria and fungi through the well diffusion method. Ketones and carboxylic acids were the main essential oil constituents. The highest total phenolic and flavonoid content as well as the best antioxidant capacity were noticed on the n-butanol extract. All the extracts showed a greater efficiency than acarbose in the inhibition of α-amylase. On the other hand, they demonstrated a mild inhibition effect against Staphylococcus aureus and Fusarium oxysporum.

Factors controlling phytoplankton dynamics in an arid reservoir in Tunisia (case of Sidi Saad dam).

Temporal variation of the phytoplankton community and environmental factors were investigated over 8 months: from January 2018 to November 2018 in the Sidi Saad reservoir (central Tunisia): May, June (spring season), July, August (summer season) September, October and November (autumn season), and January (winter season). The relationships between phytoplankton and environmental factors were explored using Canonical Correspondence Analysis (CCA). General linear models (GLMs) were used to predict the phytoplankton abundance. Analysis of variance (ANOVA) was used to test the hypothesis that the abundance of each group of taxa differed between sampling months. The results of chemical analysis of the reservoir showed that the environment was enriched in nitrates, nitrites, ammonium, and orthophosphate, especially in May. Carlson's trophic index using average chemical variables showed that Sidi Saad has a mesotrophic statute. There is temporal phytoplankton succession in the Sidi Saad reservoir. Cyanophyceae dominated over the year except in winter and late autumn (November and January). Chlorophyceae was the dominant group in winter month. The CCA results showed that six environmental factors, orthophosphates (PO43-), total phosphates (TP), Secchi disc depth (SD), total nitrogen (TN), temperature (T), and nitrite (NO2-) had significant influences on the changes in phytoplankton. GLM showed that PO43-, TP, TN, SD, and T were the significant predictors of phytoplankton abundance. Phytoplankton composition is largely dominated by the species Microcystis aeruginosa which formed a bloom with excessive abundance (up to 89.76 billion cell l-1 in spring). We recommend banning the fishing and their consumption during the period of Microcystis bloom and installing a system of biomonitoring of the levels of toxins in the water.

A Bayesian network approach to determine environmental factors controlling Karenia selliformis occurrences and blooms in the Gulf of Gabès, Tunisia.

A Bayesian Network modeling framework is introduced to explore the effect of physical and meteorological factors on the dinoflagellate red tide forming Karenia selliformis in various sampling sites of the national phytoplankton monitoring program. The proposed models took into account the physical environment effects (salinity, temperature and tide amplitude), meteorological constraints (evaporation, air temperature, insolation, rainfall, atmospheric pressure and humidity), sampling months and sites on both Karenia selliformis occurrences and blooms. The models produced plausible results and enabled the identification of the factors that directly impacted on the species occurrences and concentration levels. The sampling sites dominated the species occurrences. The models show that the relationship between salinity and Karenia selliformis is more apparent when the species concentrations are focused on and that the bloom occurrences can be predicted based on salinity. Concentrations up to 105 cells L-1 were recorded when salinity exceeded 42.5 and dominated the shallow and weak water renewal areas.

Ostreopsis cf. ovata in the Gulf of Gabès (south-eastern Mediterranean Sea): morphological, molecular and ecological characterization.

In the last few decades, the frequency of the toxic benthic dinoflagellate Ostreopsis cf. ovata proliferation has increased in the Mediterranean Sea. These blooms are associated with harmful effects on human health and the environment. The present work provides the first long term study on the spatio-temporal distribution of O. cf. ovata in relation to physical parameters in the Gulf of Gabès coastal waters (south-eastern Mediterranean Sea), as well as its morphological, molecular and physiological features. The strains of O. cf. ovata were identified morphologically by light and epifluorescence microscopy. The morphology and the size range of cultured strains were similar to those described regarding O. cf. ovata isolated from the Mediterranean Sea. The ultrastructural analysis of O. cf. ovata cells using the transmission electron microscopy showed the presence of numerous vesicles (VE) containing spirally coiled fibers (SCFs) connected to the mucus canal (CH). The phylogenetic tree based on the internal transcribed spacer region containing the 5.8S rDNA (ITS-5.8S rDNA) revealed that O. cf. ovata strains were placed into the Mediterranean/Atlantic clade. In addition, O. cf. ovata toxicity was evaluated by the mouse bioassay and a dose level≥4×104 cells was found to be lethal to mice. The examination of the O. cf. ovata occurrence in the Gulf of Gabès at a large temporal scale (1997-2012) revealed a clear seasonal pattern with dominance from midsummer (July) to late autumn (November). Furthermore, a positive correlation was found between the abundance of O. cf. ovata and salinity, whereas no correlation was found as regards temperature. The occurrence of O. cf. ovata was only detected at salinity above 35 and the highest concentrations were observed at 45. Laboratory experiments confirmed such a result and showed that isolated O. cf. ovata strains had optimal growth at salinity ranging between 35 and 45, with its peak at 40.

A Qualitative Evidence of the Breeding Sites of Anopheles arabiensis Patton (Diptera: Culicidae) in and Around Kassala Town, Eastern Sudan.

Anopheles arabiensis Patton (Diptera: Culicidae) is considered the most efficient malaria vector in eastern Sudan. This study aims to characterize the breeding sites of An. arabiensis throughout the year in and around Kassala town, eastern Sudan. Diverse larval habitat types were visited and characterized based on the habitat type and chemical composition. Mosquito larvae were found in many diverse habitats. During the rainy season, rain pools and water bodies created by the seasonal Gash River serve as the main breeding sites. In the dry season, irrigation canals, seepage from water pipes, neglected wells, artificial containers, and man-made ditches serve as the main breeding sites. Breeding water showed a pH of 7.9 and a low concentration of the total dissolved salts. The results of this study may be considered in planning and implementing larval control programs in the area.

Succession of Phytoplankton and Zooplankton Communities Coupled to Environmental Factors in the Oligo-mesotrophic Nabhana Reservoir (Semi Arid Mediterranean Area, Central Tunisia).

Ikbel Sellami, Asma Hamza, Monia El Bour, Mohammed Alaoui Mhamdi, Bernadette Pinel-alloul, and Habib Ayadi (2016) The occurrence of plankton seasonal dynamics in Mediterranean shallow lakes is characterised by a marked interannual variability, which makes it difficult to establish reliable predictions on the dynamics and functioning of plankton in these ecosystems based on the Plankton Ecology Group (PEG) model. In the present paper we study the succession of the phytoplankton and zooplankton communities in the semi-arid Mediterranean Nabhana reservoir (Central Tunisia) and its relationships with environmental factors during the period from May 2005 to January 2006 in the deepest area of this oligo-mesotrophic reservoir. Water temperature was a keystone factors in the seasonal dynamics of zooplankton. Cyanobacteria dominated the phytoplankton community throughout the study year 2005-2006 (94-99%) and Microcystis aeruginosa contributed for 93% of the total cyanobacteria abundance. The zooplankton community was dominated by copepods (66%) and cladocerans (28%). The dominant species was Copidodiaptomus numidicus (66% of total zooplankton) followed by Diaphanosoma brachyurum (22% of the total zooplankton). In addition, our results showed that the peak of phytoplankton abundance (5.6 ×106 cells l-1, June) coincided with that of zooplankton abundance (2.1 ×102 ind l-1, June) contrary to the clear-water phase model. The cyanobacteria and copepods were responsible for these exceptional peaks that occurred in June, accounting respectively for 99% of the total phytoplankton and 82% of the total zooplankton. In the Nabhana reservoir, the trophic relationship between phytoplankton and zooplankton were implicated in the phytoplankton abundance and dynamics. The occurrence of cyanobacterial blooms is determined by biotic and abiotic factors. The dynamics between cyanobacteria and their grazers may emerge as a regulator of blooms.

Bathymetric variation of epiphytic assemblages on Posidonia oceanica (L.) Delile leaves in relation to anthropogenic disturbance in the southeastern Mediterranean.

A survey of the epiphytic leaves of Posidonia oceanica was conducted along a depth transect at both the control station Attaya in the Kerkennah Islands and the disturbed Mahres station on the Sfax coast (Tunisia). Samples were collected by scuba divers at depths of 5, 10, 15, and 20 m in July 2008. We evaluated whether the pattern of spatial variability of the macroepiphyte assemblages of leaves of Posidonia oceanica differed in relation to anthropogenic interference. The results indicate that the decrease in shoot density and leaf length according to depth was low at Mahres. The biomass of epiphytic leaves and the percentage cover of epiphytic assemblages decreased with depth for both stations and heavily at Mahres, this decline being related to anthropogenic disturbance. This study shows that the highest values of epifauna and epiflora were detected at the disturbed station Mahres. Macroalgae assemblages decreased with depth at both stations and were dominated by Rhodophyta, whereas the percentage cover of the epifauna leaf that decreases according to depth was dominated by Hydrozoa and Bryozoa. Changes in epiphyte assemblages, epiphytic biomass, percentage cover, and species richness in proportion to Heterokontophyta, Rhodophyta, Cyanobacteria, Hydrozoa, Porifera, and Tunicata between the two stations constitute promising tools for detecting environmental disturbance.

Fluctuating asymmetry in grass goby Zosterisessor ophiocephalus Pallas, 1811 inhabiting polluted and unpolluted area in Tunisia.

Fluctuating asymmetry, a random deviation of bilateral traits, is assessed on grass goby Zosterisessor ophiocephalus Pallas, 1811 living in one disturbed site and in one unpolluted site Gulf of Gabes (Tunisia). Statistical analysis showed that assessed asymmetries are fluctuating. FA of orbital diameter, sub-orbital distance, pectoral fin and post-orbital length are higher in polluted site compared to control. Chemical pollution seems to be responsible for this high FA.

Long-term microphytoplankton variability patterns using multivariate analyses: ecological and management implications.

Microphytoplankton data along the southern Tunisian coasts have been monitored weekly since 1995. This study used the data collected during the period 2000-2007 to determine the temporal variability patterns and to characterize the geographical structures of microphytoplankton populations. The methodological approach consisted in isolating the temporal variability common to all sampled sites and in analyzing the "residual" site-specific component. Multivariate ordination methods were used to determine the seasonal and interannual variability. The temporal pattern shared by all sites was highlighted and the seasonal cycle of the most frequent phytoplankton communities contrasted the diatoms winter-spring species to the dinoflagellates summer-fall species. Multitable comparison allowed the identification of three areas exhibiting similar phytoplankton population variability. The results, despite the coastal location of sampling sites, indicated that hydrodynamic and geomorphologic properties of the different areas as well as anthropogenic activities play a key role in the structure of the phytoplankton communities.

Seasonal variation of Sarpa salpa fish toxicity, as related to phytoplankton consumption, accumulation of heavy metals, lipids peroxidation level in fish tissues and toxicity upon mice.

The aim of this work was to investigate for Sarpa salpa the seasonal trend in the food sources, heavy metals bioaccumulation and the oxidative stress in the organs. In addition, the toxicity was assessed by mouse bioassay of extract of the fish's organs collected in autumn, the peak of occurrence of hallucinatory syndrome. The toxicity was further studied for compounds present in epiphyte collected from the sea at the end of spring and in summer that are digested by the S. salpa in these seasons. We observed a higher lipid peroxydation in different tissues of S. salpa compared to the control fish Diplodus annularis. Furthermore, heavy metals accumulation in organs of these fish showed a significant variation between the two species (P < 0.05). The lethal dose (LD50%) determined for crude ciguatoxin (neurotoxins) extracts of viscera, liver, brain and muscle of S. salpa were as follows: 1.217, 2.195, 14.395, 18.645 g/kg mouse, respectively. We noticed a significant correlation (P < 0.05) between the total amount of toxic dinoflagellates and the level of TBARS in the liver, the brain and the muscle, this for all seasons and all sizes. Moreover, the cytotoxic effect observed for epiphytes extract confirms the transfer of toxins originating from toxic dinoflagellates, which live as epiphytes on P. oceanica leaves, to the fish by grazing. Our work indicates that, toxic phytoplanktons and heavy metals accumulation are responsible for the increase of oxidative stress in the organs of S. salpa. Hence, the edible part of S. salpa, especially the viscera and liver, can cause a threat to human health, and consumption should, for this reason, be dissuaded.

Evaluation of cytotoxic compounds in different organs of the sea bream Sarpa salpa as related to phytoplankton consumption: an in vitro study in human liver cell lines HepG2 and WRL68.

The present study was aimed to assess the cytotoxic effects of not-yet identified compounds present in organ extracts of Sarpa salpa, collected in autumn, the period with a peak in health problems. In addition, we studied the cytotoxicity of extracts of epiphytes found in the stomach content of S. salpa collected in summer and of epiphytes collected from the sea in the Sfax area at the end of spring. We tested these fractions in two human hepatic cell lines: HepG2 and WRL68. We observed a significant loss of viable cells when HepG2 cells were exposed for 72 h to acetone extracts of livers of S. salpa at a concentration of 2.5 mg/ml protein. Proteins extracted from brain or muscle did not significantly induce cell death at the studied concentrations (≤10 mg/ml). There was a significant loss of viable cells when treated with liver extract of S. salpa dissolved in DMSO. Extracts of epiphytes collected in late spring showed a cytotoxic effect in a concentration-dependent manner. Moreover, we observed a significantly decreased cell viability of HepG2 at a dilution (1/40) of epiphyte extracts from stomach contents of two fish we had collected. The cytotoxic effect of the observed epiphyte extracts confirms the transfer of toxins originating from toxic dinoflagellates which live in epiphyte on the Posidonia oceanica leaves to fish organs by grazing. Hence, the liver of this fish can cause a threat to human health and consumption should for this reason be dissuaded.

Variability in the structure of epiphytic assemblages of Posidonia oceanica in relation to human interferences in the Gulf of Gabes, Tunisia.

In this study we evaluate whether the pattern of spatial variability of the macro-epiphyte assemblages of leaves of Posidonia oceanica differed in relation to anthropogenic interference in the Gulf of Gabes (southern coast of Tunisia). A hierarchical sampling design was used to compare epiphytic assemblages at 5 m depth in terms of abundance and spatial variability at disturbed and control locations. The results indicate that the biomass and mean percentage cover decreased at locations near the point of sewage outlet in comparison to control locations. These losses were related to the distance from the source of disturbance. This study revealed that the diversity is reduced in disturbed locations by the loss of biomass and the mean percentage cover, explained by means of a multiple-stressor model which plays an important role in the macro-epiphytes' setting. It is urgent to propose the best management plans to save the remaining P. oceanica meadow in the Gulf of Gabes and its associated epiphytes.

Quantitative determination of gymnodimine-A by high performance liquid chromatography in contaminated clams from Tunisia coastline.

Quantitative determination by high performance liquid chromatography (HPLC) was performed for gymnodimine-A (GYM-A), a phycotoxin responsible for the contamination of Tunisian clams. This study demonstrates a rapid and reproducible HPLC-ultraviolet (UV) method for extraction, detection and quantification of GYM-A in toxic clams. The extraction of GYM-A from the digestive gland of clams in acetone, subsequent clean-up with diethyl ether and extraction with dichloromethane is the more valid protocol. Chromatography analyses were performed using a gradient of acetonitrile-water (10:90 to 90:10), containing trifluoroacetic acid (0.1%) for 20 min at 1 mL/min rate with a C18 column. Recovery rates exceeded 96%, and limits of detection and quantification were 5 ng/mL and 8 ng/g digestive gland, respectively. Repeatability and reproducibility were tested for various samples containing different levels of GYM-A. A significant correlation was observed between toxicity level of samples and the determined amount of GYM-A. Also, the persistence of GYM-A in contaminated clams from Boughrara lagoon was demonstrated. The kinetics discharge study of GYM-A in controlled medium, during 1 month, showed that the process of depuration was biphasic with an exponential discharge of 75% of the total amount of sequestered GYM-A during the first 12 days followed by a slow discharge (>10%) for the subsequent days up to the seventeenth day. This is the first time that a quantitative study of GYM-A in clams from Tunisian coasts is performed through the development of a new method for detection and quantify of this phycotoxin. We found HPLC-UV a reliable and suitable alternative to the mouse bioassay.