Effects of rare earth element samarium doped zinc oxide nanoparticles on Mytilus galloprovincialis (Lamarck, 1819): Filtration rates and histopathology.

BACKGROUND: Doping was reported to improve the photo catalytic performance, antioxidant, antibacterial and other biological properties of nanoparticles. While, improving the nanoparticle properties, doping could change toxicity profile to living organism. Hence, the aim of this work was to assess the effects of samarium doped zinc oxide nanoparticles (Sm doped ZnO NPs) on the edible mussel Mytilus galloprovincialis. METHODS: Sm doped ZnO nanoparticles were characterized using X-ray diffraction (XRD), transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FTIR) techniques. 156 mussels were exposed during 7 days to a low, intermediate and high concentration of Sm doped ZnO NPs (0.5, 1 and 1.5 mg/L, respectively). The filtration rates were assessed after 1 and 2 h. Histopathological alterations were determined in gills, digestive glands and gonads using a quantitative analysis. RESULTS: The filtration rates decreased in all individuals exposed to Sm doped ZnO NPs, a significant decrease was noted with the low and intermediate concentration (0.5 and 1 mg/L) of Sm doped ZnO NPs after 1 and 2 h, respectively. The histopathological index (Ih) estimated for gills, digestive glands and gonads showed differences depending on the organ and the nanoparticle concentration. The highest Ih were reported for digestive glands and female gonads exposed to the intermediate concentration (1 mg/L) of Sm doped ZnO NPs. As for gills and male gonads, the highest Ih were noted with the high concentration (1.5 mg/L) of Sm doped ZnO NPs. CONCLUSION: Results from this study revealed the toxicity of Sm doped ZnO NPs in Mytilus galloprovincialis gills, digestive glands and gonads. The toxicity induced by this nanoparticle varies depending on the organ and the concentration.

Impact of polyethylene microplastics on the clam Ruditapes decussatus (Mollusca: Bivalvia): examination of filtration rate, growth, and immunomodulation.

The present study was conducted to assess, for the first time, the effects of a 14 days experimental exposure to polyethylene (PE) based MPs (40-48 µm) on the clam Ruditapes decussatus. Clams were exposed to three different concentrations of MPs in controlled laboratory conditions: 10 µg/L (low), 100 µg/L (medium), and 1000 µg/L (high). The effects of MPs were assessed using a multi-marker approach, including the filtration rate, growth, and the integrity of immune cells (such as haemocyte numbers, viability, and lysosomal membrane destabilization). The results revealed that as the concentration of PE-MPs increased, the filtration rate decreased, indicating that PE-MPs hindered the clams' ability to filter water. Furthermore, there was a noticeable decrease in the overall weight of the clams, particularly in the group exposed to 1000 µg/L. This decrease could be attributed to the impairment of their nutrient filtration function. In terms of immune system biomarkers, exposure to PE-MPs led to immune system disruption, characterized by a significant increase in the number of haemocytic cells, especially in the group exposed to the high concentration. Additionally, there was a notable reduction in the viability of haemocytes, resulting in the destabilization of their lysosomal membranes, particularly in the groups exposed to medium and high PE-MPs concentrations. The findings of this study indicate that the sensitivity of hemolymph parameter changes and filtration rate in R. decussatus exposed to PE-MPs (100 and 1000 µg/L), surpasses that of growth performance and can serve as reliable indicators to assess habitat conditions and contaminant levels.

Acute toxicity and teratogenicity of carbaryl (carbamates), tebufenpyrad (pyrazoles), cypermethrin and permethrin (pyrethroids) on the European sea bass (Dicentrarchus labrax L, 1758) early life stages.

The toxicity of carbaryl, tebufenpyrad, cypermethrin and permethrin was evaluated in European sea bass Dicentrarchus labrax during the embryonic and larval development using six different concentrations per chemical. The order of the toxicity effectiveness was carbaryl > tebufenpyrad > cypermethrin > permethrin. The larvae were more sensitive to all tested chemicals than embryos. The LC50 of carbaryl, tebufenpyrad, cypermethrin and permethrin was determined as 13.88, 43.96, 92 and 142 ppm and 9.27, 25.67, 48.4 and 72.7 ppm in embryo and larvae, respectively. Furthermore, the tested pesticides exhibited teratogenic effects on D. labrax embryo-larval stages. The observed malformations were coagulation, no spherical egg, unhatched egg, pericardial oedemata, yolk oedemata, lordosis, kyphosis, scoliosis, no eye, cranial deformation and body atrophy. Malformations were induced with 0.5 ppm carbaryl, 10 ppm tebufenpyrad and 50 ppm cypermethrin and permethrin; the highest rates of malformation were noted with 16 ppm carbaryl, 160 ppm tebufenpyrad, 400 ppm cypermethrin and 400 ppm permethrin as 34.5%, 28%, 17.5% and 16%, respectively. A positive correlation between the incidence of malformation and the increase of pesticide concentration was established.

Monitoring of coastal pollution using shell alterations in the false limpet Siphonaria pectinata.

Lipid peroxidation level (LPO), shell biometry, shape, elemental content, and microstructure were studied in three populations of Siphonaria pectinata in the complex lagoon-channel of Bizerte across a coastal pollution gradient (northern Tunisia). LPO was found in higher concentrations in harbour populations, and shells had centred apex and were flattened. Shells were also thicker, particularly in the inner layer, with many fibrous inter-beds formed. Difference in crystallization pattern was observed in numerous shells from all three populations, being more common in harbours. From the control station to the contaminated stations, shell elemental changes were observed, with a decrease in Ca, P, Sr, and S and an increase in Cl, Cd, Cu, Fe, and K. All of these findings suggested that shell alterations could be used as a good biomarker for coastal contamination.

Effects of environmentally relevant levels of polyethylene microplastic on Mytilus galloprovincialis (Mollusca: Bivalvia): filtration rate and oxidative stress.

The objective of the present study was to evaluate the potential toxic effects of polyethylene microplastics (PE-MPs) (40-48 µm) on the Mediterranean mussel Mytilus galloprovincialis in controlled laboratory conditions. The exposure was carried out for 14 days with three environmentally relevant PE-MPs concentrations of 1, 10, and 100 and a high concentration of 1000 µg/L. Effects of PE-MPs were assessed by evaluating the filtration rate (FR) after 7 and 14 days of exposure and by analyzing biochemical biomarkers of oxidative stress (catalase - CAT, glutathione S-transferase - GST, and the levels of lipid peroxidation - LPO) in the M. galloprovincialis digestive gland after 14 days of exposure. Results showed that M. galloprovincialis does not accumulate PE-MPs of 40-48 µm size in its whole tissues. The filtration rate was significantly reduced with the increase of PE-MPs concentrations. The biochemical biomarkers indicated that PE-MPs induced oxidative damage (LPO) at low concentrations (1 and 10 µg/L) with a significant reduction in females of 1000 µg/L treated group and inactivate antioxidative system (CAT and GST) in the digestive gland of both sexes at high concentrations (100 and 1000 µg/L). This study demonstrates that PE-MPs have biological effects on M. galloprovincialis at environmentally relevant concentrations thus brings new insights on the potential impacts of PE-MPs in marine bivalves.

Microplastics in commercial molluscs from the lagoon of Bizerte (Northern Tunisia).

Microplastic (MP) pollution was investigated, for the first time, in six commercial molluscs collected from the lagoon of Bizerte during March 2018. The objective of this study was to determine the bioavailability of MPs to marine organisms and their risk for consumers of seafood. MP concentrations varied from 703.95 ±â€¯109.80 to 1482.82 ±â€¯19.20 items kg-1 wet weight. Three types of coloured MPs, including fibres, fragments and films were recovered. Fibres were the most common MP type isolated in each species. The most common size class was 0.1-1 mm. The FTIR-ATR analysis confirmed the presence of two polymer types polyethylene and polypropylene. Our results suggest that MP pollution was widespread and exhibited a relatively high level in commercial molluscs collected from Bizerte lagoon, suggesting trophic transfer in the food web and human exposure risks by diet. More investigations on MPs should be conducted in seafood and other marine organisms.

The hidden side of a major marine biogeographic boundary: a wide mosaic hybrid zone at the Atlantic-Mediterranean divide reveals the complex interaction between natural and genetic barriers in mussels.

The Almeria-Oran Front (AOF) is a recognised hotspot of genetic differentiation in the sea, with genetic discontinuities reported in more than 50 species. The AOF is a barrier to dispersal and an ecological boundary; both can determine the position of these genetic breaks. However, the maintenance of genetic differentiation is likely reinforced by genetic barriers. A general drawback of previous studies is an insufficient density of sampling sites at the transition zone, with a conspicuous lack of samples from the southern coastline. We analysed the fine-scale genetic structure in the mussel Mytilus galloprovincialis using a few ancestry-informative loci previously identified from genome scans. We discovered a 600-km-wide mosaic hybrid zone eastward of the AOF along the Algerian coasts. This mosaic zone provides a new twist to our understanding of the Atlantic-Mediterranean transition because it demonstrates that the two lineages can live in sympatry with ample opportunities to interbreed in a large area, but they hardly do so. This implies that some form of reproductive isolation must exist to maintain the two genetic backgrounds locally cohesive. The mosaic zone ends with an abrupt genetic shift at a barrier to dispersal in the Gulf of Bejaia, Eastern Algeria. Simulations of endogenous or exogenous selection in models that account for the geography and hydrodynamic features of the region support the hypothesis that sister hybrid zones could have been differentially trapped at two alternative barriers to dispersal and/or environmental boundaries, at Almeria in the north and Bejaia in the south. A preponderantly unidirectional north-south gene flow next to the AOF can also maintain a patch of intrinsically maintained genetic background in the south and the mosaic structure, even in the absence of local adaptation. Our results concur with the coupling hypothesis that suggests that natural barriers can explain the position of genetic breaks, while their maintenance depends on genetic barriers.

First assessment of the effectiveness of the international convention on the control of harmful anti-fouling systems on ships in Tunisia using imposex in Hexaplex trunculus as biomarker.

The global ban on the use of tributyltin (TBT) forced in September 2008 was ratified in Tunisia as late as June 2011. In this context, the present study aims to highlight the consequences of TBT contamination by monitoring 22 Tunisian sites before (2007) and after the ban (2012 and 2016) using as biomarker the occurrence of imposex in Hexaplex trunculus. From 2004 to 2016, complete recovery from imposex was reported in 11 sites. All average imposex indices calculated for each sampling year also decreased: imposex incidence from 61 to 27%, VDSI from 2 to 0.7, and RPLI from 15.7 to 1.6%. Ecological Quality Ratios (EQR) revealed that the sampling sites are in moderate to good ecological status. Overall, the present study confirms the effectiveness of the enacted legislation in reducing the impact of TBT pollution along the Tunisian coast.

Unexpected mosaic distribution of two hybridizing sibling lineages in the teleplanically dispersing snail Stramonita haemastoma suggests unusual postglacial redistribution or cryptic invasion.

Molecular approaches have proven efficient to identify cryptic lineages within single taxonomic entities. Sometimes these cryptic lineages maybe previously unreported or unknown invasive taxa. The genetic structure of the marine gastropod Stramonita haemastoma has been examined in the Western Mediterranean and North-Eastern Atlantic populations with mtDNA COI sequences and three newly developed microsatellite markers. We identified two cryptic lineages, differentially fixed for alternative mtDNA COI haplogroups and significantly differentiated at microsatellite loci. The mosaic distribution of the two lineages is unusual for a warm-temperate marine invertebrate with a teleplanic larval stage. The Atlantic lineage was unexpectedly observed as a patch enclosed in the north of the Western Mediterranean Sea between eastern Spain and the French Riviera, and the Mediterranean lineage was found in Macronesian Islands. Although cyto-nuclear disequilibrium is globally maintained, asymmetric introgression occurs in the Spanish region where the two lineages co-occur in a hybrid zone. A first interpretation of our results is mito-nuclear discordance in a stable postglacial hybrid zone. Under this hypothesis, though, the location of genetic discontinuities would be unusual among planktonic dispersers. An alternative interpretation is that the Atlantic lineage, also found in Senegal and Venezuela, has been introduced by human activities in the Mediterranean area and is introgressing Mediterranean genes during its propagation, as theoretically expected. This second hypothesis would add an additional example to the growing list of cryptic marine invasions revealed by molecular studies.

Imposex development in Hexaplex trunculus (Gastropoda: Caenogastropoda) involves changes in the transcription levels of the retinoid X receptor (RXR).

In order to further demonstrate that TBT-induced imposex through RXR signaling is not species-specific, Hexaplex trunculus was selected as an experimental model species. We first isolated RXR in H. trunculus, and determined gene transcription through quantitative real-time PCR in key tissues (e.g., penis/penis-forming area and central nervous system:- CNS), upon exposure to tributyltin (TBT) (5 and 50 ng TBTL(-1)). Two months of exposure to TBT induced imposex and led to a significant increase in the severity of the phenomenon in females and an increase in male penis lengths. Exposure to TBT altered RXR gene transcription in a tissue and sex-specific manner. In the CNS, there were no significant changes in RXR gene transcription between control and TBT-exposed females. A similar trend was observed in male CNS. On the contrary, in the penis-forming area/penis of females exposed to TBT, a significant increase in RXR gene transcription was observed in the 50 ng TBTL(-1) group. Furthermore, a positive correlation was observed between overall female penis lengths and RXR gene transcription. In males, although a trend towards an increase in RXR gene transcription in penis was observed, differences did not reach significance. Overall, the results of the present study give further support to a local role of RXR in the penis-forming area during the development of imposex by TBT, thus suggesting a conserved function of RXR in penis formation at least within prosobranch gastropods.

Imposex and butyltin concentrations in Bolinus brandaris (Gastropoda: Muricidae) from the northern Tunisian coast.

The imposex incidence and butyltin concentration i.e. tributyltin with its di- and mono-substituted metabolites were investigated in the muricid Bolinus brandaris sampled from two sites on the northern Tunisian coast (the Lagoon of Bizerta and the small Gulf of Tunis). Both populations had imposex, with stages of imposex development varying between VDS 1 and VDS 4.3. All imposex indices (imposex frequency (I %), female penis length, female vas deferens length, vas deferens sequence index, relative penis length index, and vas deferens length index) were significantly higher in snails from the Bizerta lagoon. Butyltins were detected in the whole tissues of both sexes from the two sites. TBT levels were higher in gastropods collected from the lagoon of Bizerta (12.65 ±1.48 ng Sn g( - 1) dw in female and 15.21 ±1.13 ng Sn g( - 1) dw in male) than in individuals from the Gulf of Tunis (10.71 ±1.26 ng Sn g( - 1) dw in female and 11.65 ±1.63 ng Sn g( - 1) dw in male), corroborating the data of imposex analysis. These results confirmed that B. brandaris could be used as a bioindicator species of butyltin pollution in the studied areas. In addition, this study provided baseline data that could serve for long-term monitoring of TBT pollution in Tunisia, since legislation to reduce the use of TBT-based antifouling paints has not been introduced yet.

Potential of Nassarius nitidus for monitoring organotin pollution in the lagoon of Bizerta (northern Thunisia).

Imposex and butyltin burden were assessed in Nassarius nitidus, Bolinus brandaris and Hexaplex trunculus collected at five stations in the Bizerta lagoon. Biological analysis showed that imposex followed type (a) in N. nitidus (distal evolution), against type (d) in the two muricids (proximal evolution). Imposex indices were higher in sites located nearby sources of tributyltin and N. nitidus was the least affected species of the five sites, followed by B. brandaris and H. trunculus. Butyltin analysis showed lower accumulation in N. nitidus followed by H. trunculus and B. brandaris. This study has allowed the gathering of data on imposex in a snail studied for the first time in Tunisia (N. nitidus). It suggests the possibility of using such snail as a complementary species for organotin monitoring programs in the Mediterranean and further confirmed that H. trunculus is the most suitable species for such investigations.