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Abstract The garfish Belone belone represents the only valid endemic Belone species for the Mediterranean Sea and the eastern Atlantic Ocean. It shows a wide global distribution range, with a high commercial value and ecological relevance in the pelagic domain. Despite this, there needs to be more knowledge regarding the otoliths of this species, with the total absence of descriptions regarding asterisci and lapilli from Mediterranean populations and a lack of studies on the reliability of shape analysis on its sagittae. The present paper aims to provide the first main contours description of the three otoliths pairs from a Mediterranean population, providing an accurate investigation of morphology, morphometry, and intra-specific variability of sagittae, lapilli, and asterisci. Results showed (i) the absence of directional bilateral asymmetry and sexual asymmetry for the three otoliths pairs, (ii) a different morphology and morphometry of sagittae, lapilli and asterisci than those described in the literature, and (iii) an enhanced variability between sagittae morphometry and shape between the three investigated size classes. All these data confirmed the reliability of the studied species of shape analysis, showing a geographical and size-related variability of otoliths features probably related to genetics, environmental conditions, and life habits variations.

Improve the knowledge base on the ecomorphological adaptation of teleost species to different environments, trying to reconstruct how habitat can shape sagittae, is essential for conservational purposes, evolutionary evaluations, and population dynamics studies. Here is provided a comparative study between sagittae features, growth rates and age composition of two Scorpaena porcus populations inhabiting the Mediterranean waters, one from the Strait of Messina (Central Mediterranean Sea) and one from the Split area (Adriatic Sea). Additionally, the stomach contents of the Messina specimens have also been analysed to provide some preliminary information on the diet of S. porcus. A total of ninety individuals, half from each area, have been obtained from two extremely different environments. Results showed an overall different morphology, shape, and morphometry of sagittae among the size classes of the two investigated populations. Samples from Messina were characterized by a most elliptical and slender shape, with a more regular serration of margins than those from Split, which exhibited a wider sagitta, with a most enhanced anti-rostrum and longer rostrum. Concerning diet, specimens from Messina showed a preference for Crustacea (especially Brachyura and Amphipoda) and Teleost species, showing some differences with literature data from other geographical areas (Split one included). Results have confirmed the reliability of sagittae to detect the inter-population variability of S. porcus from different geographical areas, an essential tool for stock assessment, population studies and investigation on ecomorphological adaptation of teleost species to different habitats.

The journal retracts the article, “Assessment of 2-Pentadecyl-2-oxazoline Role on Lipopolysaccharide-Induced Inflammation on Early Stage Development of Zebrafish (Danio rerio)” [...]

The ubiquitous presence of microplastics (MPs) in the environment is a major concern for the conservation of biodiversity. Numerous studies show the detrimental effects of MPs on marine species, especially because of their small size and their capacity to absorb organic contaminants from their surroundings. The bivalve mollusk Cerastoderma glaucum (Bruguière, 1789), because of its wide geographic distribution and immobile feeding habits, can be used as a sentinel and bioindicator species. By examining the presence, localization, and co-localization of Toll-like receptor 2 (TLR2) and inducible nitric oxide synthetase (iNOS), this study aims to evaluate the response of the internal defense system of C. glaucum to pristine MPs through the employment of confocal microscopy and bioinformatics techniques. The results show haemocytes immunoreactive to the antibodies tested; in particular, a higher number of TLR2-positive haemocytes can be observed in the group exposed to pristine MPs. These findings suggest that haemocytes can play a key biomarker role as sentinels to environmental pollutants. Furthermore, bioinformatics analyses on the antibodies tested confirmed an evolutionary conservation of these molecules. These data highlight the critical role of phagocytosis in identifying ecosystem damage and are helpful in developing biosensors with less negative effects on the environments in which they are applied.

Cold-water corals (CWCs) are bioengineering species that can increase habitat heterogeneity and improve the deep sea’s biological diversity and ecosystem functioning. Knowledge of their distribution provides a critical baseline for assessing the effect of natural and anthropogenic impacts on these important deep-sea habitats. The aims of this study are: i) provide new data on the spatial distribution of six CWCs species in the Strait of Sicily, ii) describe the principal environmental and anthropogenic variables that play a role in shaping their distribution, iii) identify hotspots in which individuals belonging to the various species co-occur. Presence-only data of six CWCs species, ten environmental variables (depth, slope, rugosity, aspect, flowdir, temperature, salinity, north bottom current, east bottom current, chlorophyll-a), and one variable relating to bottom trawling effort (Automatic Information System – AIS) were used to predict the suitable habitats. We used Maximum Entropy modelling (MaxEnt) approach and used the AUC (area under the receiver operating characteristic curve) and TSS (true skill statistics) to evaluate the model performance. The results showed excellent AUC, TSS and AUC’s standard deviation mean values for all six species. The validation show high predictive performance. MaxEnt identified slope, depth, and rugosity as the most important predictors, showing the highest percentage contribution for all six species considered. Throughout the study area, highlyinterspecific persistent density hotspot of CWCs co-occurrence were discovered, with a total extension of 4.05 km2 where all species co-occur. Although studies on the effect of environmental and anthropogenic factors that impact the distribution of these species of conservation interest remain scarce, the results of this study offer useful guidance for decision-makers to develop necessary conservation measures.

Abstract Metazoans have several mechanisms of internal defense for their survival. The internal defense system evolved alongside the organisms. Annelidae have circulating coelomocytes that perform functions comparable to the phagocytic immune cells of vertebrates. Several studies have shown that these cells are involved in phagocytosis, opsonization, and pathogen recognition processes. Like vertebrate macrophages, these circulating cells that permeate organs from the coelomic cavity capture or encapsulate pathogens, reactive oxygen species (ROS), and nitric oxide (NO). Furthermore, they produce a range of bioactive proteins involved in immune response and perform detoxification functions through their lysosomal system. Coelomocytes can also participate in lithic reactions against target cells and the release of antimicrobial peptides. Our study immunohistochemically identify coelomocytes of Lumbricus terrestris scattered in the epidermal and the connective layer below, both in the longitudinal and in the smooth muscle layer, immunoreactive for TLR2, CD14 and α-Tubulin for the first time. TLR2 and CD14 are not fully colocalized with each other, suggesting that these coelomocytes may belong to two distinct families. The expression of these immune molecules on Annelidae coelomocytes confirms their crucial role in the internal defense system of these Oligochaeta protostomes, suggesting a phylogenetic conservation of these receptors. These data could provide further insights into the understanding of the internal defense system of the Annelida and of the complex mechanisms of the immune system in vertebrates.

Abstract Anthropogenic microparticles (AMs) were found for the first time in specimens of Trematomus bernacchii collected in 1998 in the Ross Sea (Antarctica) and stored in the Antarctic Environmental Specimen Bank. Most of the identified AMs were fibers of natural and synthetic origin. The natural AMs were cellulosic, the synthetic ones were polyester, polypropylene, polypropylene/polyester, and cellulose acetate. The presence of dyes in the natural AMs indicates their anthropogenic origin. Five industrial dyes were identified by Raman spectroscopy with Indigo occurring in most of them (55%). Our research not only adds further data to the ongoing knowledge of pollution levels in the Antarctic ecosystem, it provides an interesting snapshot of the past, highlighting that microplastics and anthropogenic fiber pollution had already entered the Antarctic marine food web at the end of the ‘90 s. These findings therefore establish the foundations for understand the changes in marine litter pollution over time.

The introduction of pollutants, such as thiacloprid and benzo[a]pyrene (B[a]P), into the waters of urbanized coastal and estuarine areas through fossil fuel spills, domestic and industrial waste discharges, atmospheric inputs, and continental runoff poses a major threat to the fauna and flora of the aquatic environment and can have a significant impact on the internal defense system of invertebrates such as mussels. Using monoclonal and polyclonal anti-Toll-like receptor 2 (TLR2) and anti-inducible nitric oxide synthetase (iNOS) antibodies for the first time, this work aims to examine hemocytes in the mantle and gills of M. galloprovincialis as biomarkers of thiacloprid and B[a]P pollution and analyze their potential synergistic effect. To pursue this objective, samples were exposed to the pollutants, both individually and simultaneously. Subsequently, oxidative stress biomarkers were evaluated by enzymatic analysis, while tissue changes and the number of hemocytes in the different contaminated groups were assessed via histomorphological and immunohistochemical analyses. Our findings revealed that in comparison to a single exposure, the two pollutants together significantly elevated oxidative stress. Moreover, our data may potentially enhance knowledge on how TLR2 and iNOS work as part of the internal defense system of bivalves. This would help in creating new technologies and strategies, such as biosensors, that are more suitable for managing water pollution, and garnering new details on the condition of the marine ecosystem.

Mesopelagic species are enjoining increasing attention due to the growing impact of fisheries activities on deep marine biocenosis. Improving the knowledge base on mesopelagic species is required to enhance their conservation due to the knowledge gaps regarding many species and families. In this context, otoliths can be fundamental to assessing their life history, ecomorphological adaptation to the deep environment and stock composition. The present paper aims to explore the saccular and utricular otoliths morphology and intra-specific variability of the hatchetfish, Argyropelecus hemigymnus, from the Strait of Messina. Lapilli and sagittae were collected from 70 specimens and separated into four size classes. Morphometric, shape and SEM investigations were performed to describe their morphology, contours, and external structural organization, also studying their intraspecific variability related to sample sizes and differences between otolith pairs. Results showed an otolith morphology different from those reported in the literature with fluctuating asymmetry in sagittae and lapilli belonging to Class IV, and a high otolith variability between all the size classes. Data herein described confirm the otoliths singularity of the population from the Strait of Messina, shaped by a unique marine environment for oceanographic and ecological features.

Abstract Otolith features are useful tools for studying taxonomy, ecology, paleontology, and fish biology since they represent a permanent record of life history. Nevertheless, the functional morphology of otoliths remains an open research question that is useful to completely understand their eco-morphology. This study aims to deepen the knowledge of intra- and interspecific variation in sagitta morphology in three congeneric seabreams, to understand how such variability could be related to the lifestyles of each species. Therefore, the sagittae (n = 161) of 24 Pagellus bogaraveo, 24 Pagellus acarne, and 37 Pagellus erythrinus specimens, collected from the south Tyrrhenian Sea, were analyzed using scanning electron microscopy and a stereomicroscope to assess morphometric features, variability between otolith pairs and the external crystalline structure the of sulcus acusticus. Statistical analysis demonstrated that, between the species, variability in sagittal otolith rostral length growth and sulcus acusticus features, correlated with increased fish total length and body weight. Moreover, slight differences between otolith pairs were detected in P. acarne and P. erythrinus (P

Pharmaceuticals are widely recognized as potentially hazardous to aquatic ecosystems. In the last two decades, the constant intake of biologically active chemicals used in human healthcare has been related to the growing release of these agents into natural environments. As reported by several studies, various pharmaceuticals have been detected, mainly in surface water (seas, lakes, and rivers), but also in groundwater and drinking water. Moreover, these contaminants and their metabolites can show biological activity even at very low concentrations. This study aimed to evaluate the developmental toxicity of exposure to the chemotherapy drugs gemcitabine and paclitaxel in aquatic environments. Zebrafish (Danio rerio) embryos were exposed to doses of gemcitabine 15 μM in combination with paclitaxel 1 μM from 0 to 96 h post-fertilization (hpf) using a fish embryo toxicity test (FET). This study highlights that both gemcitabine and paclitaxel exposure at single non-toxic concentrations affected survival and hatching rate, morphology score, and body length after exposure in combination. Additionally, exposure significantly disturbed the antioxidant defense system and increased ROS in zebrafish larvae. Gemcitabine and paclitaxel exposure caused changes in the expression of inflammation-related, endoplasmic reticulum stress-related (ERS), and autophagy-related genes. Taken together, our findings underline that gemcitabine and paclitaxel increase developmental toxicity in zebrafish embryos in a time-dependent manner.

The immune system of a fish has cellular and molecular defense mechanisms that are substantially retained throughout the evolution of vertebrates. The innate immune system provides biological processes, such as phagocytosis and mechanical barriers, to implement an efficient defensive response after exposure to chemical or biological contaminants, pollutants, and contact with parasites, germs, and pathogens. Club cells (CCs) are widespread in the skin of Ostariophysi. After a predator attack or exposure to toxins and parasites, these cells can produce alarming substances. Given their effectiveness against viruses, parasites, and common skin lesions, recent studies have suggested that CCs are a component of the immune system. This study aims to immunohistochemically characterize the CCs for the first time in the skin of zebrafish, using mitogen-activated protein kinase (MAPK) p38, Toll-like receptor (TLR)2, Piscidin1, and inducible nitric oxide synthase (iNOS) peptides involved in the function of all types of vertebrate immune cells. According to our analysis, the intermediate layer of the epidermis exhibited rounded, oval, and elongated CCs, with central acidophilic cytoplasm and a spherical basophilic nucleus, that are positive to the antibodies tested. Our results may confirm that CCs could be involved in the immune function, increasing our knowledge of the immune system of teleosts.

The amphibious teleost Giant mudskipper (Periophthalmodon schlosseri, Pallas 1770) inhabit muddy plains and Asian mangrove forests. It spends more than 90% of its life outside of the water, using its skin, gills, and buccal-pharyngeal cavity mucosa to breathe in oxygen from the surrounding air. All vertebrates have been found to have mast cells (MCs), which are part of the innate immune system. These cells are mostly found in the mucous membranes of the organs that come in contact with the outside environment. According to their morphology, MCs have distinctive cytoplasmic granules that are released during the degranulation process. Additionally, these cells have antimicrobial peptides (AMPs) that fight a variety of infections. Piscidins, hepcidins, defensins, cathelicidins, and histonic peptides are examples of fish AMPs. Confocal microscopy was used in this study to assess Piscidin1 expression in Giant Mudskipper branchial MCs. Our results demonstrated the presence of MCs in the gills is highly positive for Piscidin1. Additionally, colocalized MCs labeled with TLR2/5-HT and Piscidin1/5-HT supported our data. The expression of Piscidin1 in giant mudskipper MCs highlights the involvement of this peptide in the orchestration of teleost immunity, advancing the knowledge of the defense system of this fish.

Deep Sea environments represent the final collector of anthropogenic debris mainly represented by both plastic and non-plastic materials with different size. This led to potential contamination of deep marine fauna due to direct and indirect ingestion, representing a potential hazard for the species itself and for the final consumer. In this framework, the present study explored the occurrence of anthropogenic debris in the gastrointestinal tract of three Decapoda species of high commercial and ecological value (Parapenaeus longirostris, Aristeus antennatus, and Aristaeomorpha foliacea) from south-western Ionian Sea. After morphometrical measurements and sex determination, the gastrointestinal tract of 136 specimens were extracted and then chemically digested. A total of 230 low density microparticles were isolated, with a high frequency of occurrence in all the analyzed species (76% in P. longirostris, 70% in A. antennatus, and 83% in A. foliacea) mainly represented by fibers (92.6%) with a size between 0.10 and 0.49 mm, and with a dominance of the blue color. The results of the present study report for the first time the anthropogenic debris presence in the studied Decapoda from south-western Ionian Sea, highlighting the necessity to broaden the knowledge about anthropogenic debris pollution status in Mediterranean deep-sea species.

Monitoring the occurrence and distribution of rare fish species is essential, especially in a semi-closed basin such as the Mediterranean Sea. Biologically interesting species often represent fishing waste (being part of the by-catch) and therefore are not retained, leading to a huge gap in data collection. Here we report about the occurrence in the Ionian Sea of a Zu cristatus (Bonelli, 1819) specimen, a mesopelagic fish from the order Lampriformes considered rare in the whole Mediterranean basin. The sample was captured at a depth of 720 m off the coast of Noto (Sicily, Italy) by deep-sea longline fisheries. The biometrics and meristic data were compared with all the available data on specimens previously reported in the Mediterranean Sea. It is interesting to note that, among the few records in the literature, most reports mention a lower depth range for this species. In fact, it is quite known that juvenile specimens of Z. cristatus were frequently found in shallow waters, while information about the biology of the adult stage is scarce, indicating they would seem to prefer much deeper environments. Considering the difficulty to collect data on this rare fish, this contribution can help to improve the knowledge about its distribution.

Lampriformes are circumglobally distributed and contain several families of strictly marine bony fishes that have a peculiar morphology. Lampriformes systematics is affected by limitations in biometric, meristic, and molecular data; for this reason, it underwent several rearrangements in the past. This review aimed to describe the biological and ecological characteristics of the order Lampriformes, summarizing the current taxonomy of the group. The main aim was to clarify what is known about the distribution of the order Lampriformes in the Mediterranean Sea, collecting all the scarce and fragmented reports and notes on their occurrence. Knowledge scarcity is due to their solitary nature, in addition to their low to absent economic value. Despite this, the order Lampriformes represents a taxon of high biological and ecological importance. The high depth range of distribution characterizes their lifestyle. In the Mediterranean Sea, four families are present—Lampridae, Lophotidae, Regalecidae, and Trachipteridae—with the following species respectively, Lampris guttatus (Brünnich, 1788), Lophotus lacepede (Giorna, 1809), Regalecus glesne (Ascanius, 1772), Trachipterus arcticus (Brünnich, 1788), T. trachypterus (Gmelin, 1789), and Zu cristatus (Bonelli, 1819). Data deficiencies affect information on this taxon; the present review, which collected all the reports of the Mediterranean Sea, creates a baseline for depicting the biogeography of these rare and important species.

Dendritic cells (DCs) are antigen-presenting cells (APCs) that regulate the beginning of adaptive immune responses. The mechanisms of tolerance to antigens moving through the digestive tract are known to be regulated by intestinal DCs. Agnatha and Gnathostoma are descendants of a common ancestor. The Ostracoderms gave rise to Cyclostomes, whereas the Placoderms gave rise to Chondrichthyes. Sarcopterygii and Actinopterygii are two evolutionary lines of bony fishes. Brachiopterygii and Neopterygii descend from the Actinopterygii. From Neopterygii, Holostei and Teleostei evolved. Using immunohistochemistry with TLR-2, Langerin/CD207, and MHC II, this study aimed to characterize intestinal DCs, from myxines to teleosts. The findings reveal that DCs are positive for the antibodies tested, highlighting the presence of DCs and DC-like cells phylogenetically from myxines, for the first time, to teleosts. These findings may aid in improving the level of knowledge about the immune system’s evolution and these sentinel cells, which are crucial to the body’s defense.

Veterinary antiparasitic pharmaceuticals as well as pesticides have been detected in surface waters, and they may cause several toxic effects in this environmental compartment. In the present study, we evaluated the toxicity after exposure of different concentration of ivermectin (IVM; 50, 100, and 200 μg L−1) and cypermethrin (CYP; 5, 10, and 25 μg L−1) and the combination of these two compounds at non-toxic concentration (IVM 100 + CYP 5 μg L−1) in zebrafish embryos. Combination of IVM at 100 μg L−1 with CYP at 5 μg L−1 exposure induced hatching delay and malformations at 96 hpf in zebrafish larvae as well as significant induction of cell death in zebrafish larvae. At the same time, the two single concentrations of IVM and CYP did not show a toxic effect on zebrafish development. In conclusion, our study suggests that IVM and CYP show a synergistic effect at common, ineffective concentrations, promoting malformation and cell death in fish development.

Merluccius merluccius is one of the most important Mediterranean benthopelagic predators. It represents a key species for the ecosystem’s functioning due to its fundamental role in the energy transferal between different domains and depth strata. The aim of this study was to explore the feeding habits of European hakes in the southern and central Mediterranean Sea, and also to analyze timescale variations and ontogenetic shift in five size length classes. A total of 411 stomachs collected from 2018 to 2020 were analyzed to assess diet and feeding habits. Results confirmed hakes’ role as a generalist benthopelagic predator, preying both in the suprabenthic layer and in the entire water column. Concerning the ontogenetic diet shift, juvenile hakes prefer zooplanktonic prey, while larger hakes have a diet mainly based on teleosts and decapods. The variations in diet composition between years, characterized by a fluctuation of cephalopods, bioluminescent teleost species and mesopelagic crustaceans, have highlighted the ability of European hake to model its diet to the geographical and prey availability. These features make analysis of the diet of M. merluccius essential to understanding the trophic dynamic existing in bentho-meso-pelagic environments, to improve ecosystem conservation in accordance with ecosystem-based fishery management.

(1) Background: Multiple contaminations of several mycotoxins have been detected in human and veterinary food and feed worldwide. To date, a number of studies on the combined effects of mycotoxins have been conducted on cell and animal models, but very limited studies have been done on aquatic organisms. (2) The purpose of the present study was to evaluate the combined toxic effects of Aflatoxin B1 (AFB1) and Fumonisin B1 (FB1) on zebrafish (Danio rerio) embryos. (3) Results: Our results showed that the combination of AFB1 and FB1 at nontoxic concentrations exerted a negative effect on the lethal endpoints analyzed, such as survival, hatching, and heart rate. In addition, the mixture of mycotoxins caused an increase in the levels of enzymes and proteins involved in the antioxidant process, such as superoxide dismutase (SOD) and catalase (CAT), both in terms of protein levels and gene expression, as well as an increase in the levels of the detoxification enzymes glutathione s-transferases (GST) and cytochromes P450 (CYP450). Furthermore, we showed that the mycotoxin mixture induced an increase in pro-apoptotic proteins such as bax and caspase 3, and at the same time reduced the gene expression of the anti-apoptotic bcl-2 protein. Finally, a significant decrease in thyroid function was observed in terms of triiodothyronine (T3), thyroxine (T4), and vitellogenin (VTG) levels. (4) Conclusion: We can say that the mixture of mycotoxins carries a greater risk factor than individual presences. There is a greater need for effective detoxification methods to control and reduce the toxicity of multiple mycotoxins and reduce the toxicity of multiple mycotoxins in feed and throughout the food chain.

The present paper represents the first description of abnormal hermaphroditism in Galeus melastomus (Rafinesque, 1810). The black mouth cat shark specimen, collected in summer from southern Tyrrhenian Sea at the entrance of Salerno gulf, showed a basic intersexuality characterized by the presence of male secondary and female primary sexual characters. The reproductive system has been described with an accurate morphological analysis, also including a histological assessment of oocytes. Results showed the presence of only right clasper, not exceeding the pelvic fin in length, with the absence of internal males’ sexual organs. Concerning the female primary characters, the analysis of histological assessment showed the structure of mature oocyte, confirming the maturity stage of a mature female in active extruding stage, as highlighted by the macroscopic morphological analysis. Despite in many marine organisms’ hermaphroditism is a physiological condition showed by specimens during their lifecycle, in Elasmobranchs this is considered an abnormal condition, with some exceptions. The lack of information on hermaphroditism in elasmobranchs form the Mediterranean Sea requires an increase of scientific community’s attention, improving the knowledge on the reproductive biology of this sensitive taxon. This is essential to enhance the conservation of elasmobranchs populations in the entire Mediterranean basin.

Pharmaceuticals are widely acknowledged to be a threat to aquatic life. Over the last two decades, the steady use of biologically active chemicals for human health has been mirrored by a rise in the leaking of these chemicals into natural environments. The aim of this work was to detect the toxicity of sodium fluoride (NaF) exposure and platinum-derived drugs in an ecological setting on aquatic organism development. From 24 to 96 h post-fertilization, zebrafish embryos were treated to dosages of NaF 10 mg/L−1 + cisplatin (CDDP) 100 μM, one with NaF 10 mg/L−1 + carboplatin (CARP) 25 μM, one with NaF 10 mg/L−1 + CDDP 100 μM + CARP 25 μM. Fluoride exposure in combination with Cisplatin and Carboplatin (non-toxic concentration) had an effect on survival and hatching rate according to this study. Additionally, it significantly disturbed the antioxidant defense system and increased ROS in zebrafish larvae. NaF 10 mg/L−1 associated with CDDP 100 μM and CARP 25 μM, increased the production of apoptosis-related proteins (caspase 3, bax, and bcl-2) and the downregulation of acetylcholinesterase (AChE) activity, while no effect was seen for the single exposure.

Pharmaceuticals are widely regarded as a menace to the aquatic environment. The constant consumption of biologically active chemicals for human health has been matched by an increase in the leaking of these compounds in natural habitats over the last two decades. This study was aimed to evaluate the molecular pathway underling the developmental toxicity of exposure in the ecological environment. Zebrafish embryos were exposed at doses of dexamethasone sodium phosphate (DEX) 1 μmol/L, tocilizumab 442.1 μmol/L and dexamethasone + tocilizumab (1 μmol/L and 442.1 μmol/L, respectively) from 24 h post-fertilization (hpf) to 96 hpf. This study confirmed that DEX exposure in association with tocilizumab 442.1 μmol/L at 1 μmol/L (non-toxic concentration) affected the survival and hatching rate, morphology score, and body length. Additionally, it significantly disturbed the antioxidant defense system in zebrafish larvae. Furthermore, a DEX 1 μmol/L and tocilizumab 442.1 μmol/L association also increased the production of apoptosis-related proteins (caspase-3, bax, and bcl-2).

This study aims to evaluate some biochemical parameters and serum electrolytes in cultured rainbow trout Oncorhynchus mykiss (Walbaum, 1792) to evaluate a potential correlation with biometric parameters (weight and length). For this purpose, 100 cultured trout (300–700 g weight range, 25–38 cm length range) bred on a fish-farm were used for the study. Physico-chemical characteristics of water were measured on the farm. Blood samples were collected from each fish to analyze the following parameters: glucose, triglycerides, cholesterol, aspartate aminotransferase (AST); alanine aminotransferase (ALT), calcium (Ca2+), chlorine (Cl−), iron (Fe2+), phosphorus (P), magnesium (Mg2+), potassium (K+), sodium (Na+) and urea. Statistical data analysis showed a significant correlation between size and glucose and cholesterol. No correlation was found between size and other parameters studied. These results represent a contribution to the study of fish size leading to better understanding some biochemical parameters and serum electrolyte profiles in cultured rainbow trout. This research contributes to understanding the intra-individual variability of some blood parameters in cultured rainbow trout O. mykiss offering reliable information on chronic stress status, metabolic disorders and deficiencies in relation to different sizes. These could help in improve the health monitoring in trout fish farms.

Imidacloprid (IMI) is part of the neonicotinoids family, insecticides widely used by humans and also found in wastewater. This class of compounds, if present in the environment, can cause toxicity to different species such as bees and gammarids, although little is known about vertebrates such as fish. In addition, several substances have been reported in the environment that can cause damage to aquatic species, such as potassium perchlorate (KClO4), if exposed to high concentrations or for long periods. Often, the co-presence of different contaminants can cause a synergistic action in terms of toxicity to fish. In the present study, we first analyzed different concentrations of IMI (75, 100 and 150 mg/L) and KClO4 (1, 1.5 and 5 mM) to highlight the morphological effects at 96 hpf and, subsequently, chose two nontoxic concentrations to evaluate their co-exposure and the pathway involved in their co-toxicity. Morphological alteration, mucus production, messenger RNA (mRNA) expression related to intestinal function and oxidative stress were measured. These results suggest that co-exposure to IMI and KClO4 could affect zebrafish embryo development by increasing gut toxicity and the alteration of antioxidative defense mechanisms.

The increasing pollution of aquatic habitats with anthropogenic compounds has led to various test strategies to detect hazardous chemicals. However, information on the effects of pollutants on the thyroid system in fish, which is essential for growth, development, and parts of reproduction, is still scarce. Modified early life-stage tests were carried out with zebrafish exposed to the known thyroid inhibitor potassium perchlorate (0.1, 1, 1.5, 2, 2.5, and 5 mM) to identify adverse effects on embryo development. The endogenous antioxidant defense mechanism is one of the key functions of the thyroid gland; in this regard, we examined the co-exposure to potassium perchlorate (KClO4), which could disrupt thyroid function, with cadmium (Cd), a known pro-oxidant compound. Zebrafish embryos were exposed to control KClO4 1 mM and Cd 0.5 μM for 96 h after fertilization (hpf) individually and in combination. The morphological alteration, body length, and messenger RNA (mRNA) expression related to thyroid function and oxidative stress, thyroid hormone levels, and malondialdehyde were measured. Significant down-regulation of mRNAs related to thyroid function (thyroid hormone receptor-alpha (THRα), thyroid hormone receptor-beta (THRβ), haematopoietically expressed homeobox (hhex)) and decreased thyroxin (T4) levels were observed after co-exposure to KClO4 and Cd, but this was not observed in the individually treated groups. These results suggest that co-exposure to KClO4 and Cd could affect antioxidant defense mechanisms and potentially normally increase Cd toxicity on mRNA expression, altering the thyroid functions important in zebrafish embryonic developmental stages.

Several anthropogenic products in wastewater are considered a threat to the aquatic environment. In addition to common industrial pollutants, levels of pharmaceuticals have been increasingly found in the environment in recent years, which may present a strong risk to the aquatic species that live there. The constant consumption of biologically active chemicals for human health has been matched by an increase in the leaking of these compounds in natural habitats over the last two decades. This study is aimed at evaluating the developmental toxicity of fotemustine in the ecological environment. Zebrafish embryos were exposed to doses of 25, 50 and 100 µg/mL from 4 h post-fertilization to 120 h. This study confirms that fotemustine exposure at 50 and 100 µg/mL affects the survival and hatching rate, morphology score and body length. Additionally, it significantly disturbs the antioxidant defense system and increases ROS in zebrafish larvae. From the molecular point of view, fotemustine exposure strongly induces apoptosis, endoplasmic reticulum stress (ERS) and the Wnt signaling pathway.

Pharmaceuticals are actually identified as a threat to the ecosystem. Nowadays, the growing consumption of antineoplastic agents has been related to their continuous input in natural environments. These substances can interfere with physiological and biochemical processes of aquatic species over their entire life cycle. Oxaliplatin (OXA) is a widely used chemotherapeutic agent to treat colon or rectal cancer. This study was aimed to evaluate the developmental toxicity of the OXA exposure. To this end, zebrafish embryos were incubated with 0.001, 0.1, 0.5 mg/L OXA. At different timepoints mortality rate, hatching rate, developmental abnormalities, histological analysis, oxidative stress and mRNA expression of gene related to oxidative stress were evaluated. Our results showed that OXA exposure can induce increased mortality and developmental abnormalities reducing the hatching rate. Histological analysis demonstrated that OXA induced liver, intestine, muscle and heart injury. Superoxide dismutase and catalase activities were significantly increased after OXA exposure demonstrating its oxidative effects. The mRNA expression levels of apoptosis-related genes (caspase-3, bax and bcl-2) were significantly upregulated by OXA exposure. In conclusion, we highlighted that OXA exposure led to a dose-related developmental toxicity, oxidative stress and apoptosis.

Transitional water environments represent very ecologically interesting areas, which provide various ecosystem services, both concerning biodiversity protection and sustainable fruition of resources. In this way, the evaluation of total carbon and its components, chlorophyll, and chemical and physical parameters is of fundamental importance to deepen the dynamics of these peculiar natural areas. Commercial interests linked to the biological resources of these areas are often not well exploited in relation to their sustainability, due to lack of knowledge. In this study, we investigated the distribution of total organic carbon, chlorophyll, and other related physical and chemical parameters in the natural Lagoon of Capo Peloro (Eastern Sicily), to deepen the knowledge on the carbon equilibrium of these transitional basins. Collected data showed different trends for all parameters, mainly related to different seasons and water exchanges with sea. The influences of primary production sources and farmed molluscs were not negligible and deserve to be further investigated in the future. The results obtained reveal good margins for the possibility of environmentally sustainable exploitation of natural resources in both basins, but at the same time, there is a need for a more detailed knowledge of anthropogenic impacts on the area.

IBD (Inflammatory Bowel Disease) is an inflammatory disease affecting the gastrointestinal tract that is common in both humans and veterinarians. Several studies have revealed the pharmacological properties of the oxazoline of palmitoylethanolamide (PEAOXA). Zebrafish larvae were exposed to sodium dextran sulphate (DSS) to induce enterocolitis and study the protective action of PEAOXA. After repetitive exposure with 0.25% DSS, larvae presented gut alteration with an increase in mucus production. Furthermore, DSS exposure induced an increase in the inflammatory pathway in the intestine, related to an increase in the Endoplasmic-reticulum (ER) stress genes. PEAOXA exposure at a concentration of 10 mg/L decreased the DSS-induced gut damage and mucus production, as well as being able to reduce the inflammatory and ER stress-related genes expression. In conclusion, our results demonstrate that the alterations induced by repeated exposure to DSS were counteracted by PEAOXA action that was able to inhibit the increase in inflammation and ER stress involved in the progression of enterocolitis.

Lipopolysaccharide (LPS), or bacterial endotoxin, is an important virulence factor in several human and animal pathologies. Oxazoline of Palmitoylethanolamide (PEAOXA) has shown strong anti-inflammatory activity in several animal models. LPS was applied for 24 h to zebrafish embryos to induce inflammation, and then the anti-inflammatory action of PEAOXA was evaluated for the first time in the zebrafish model (Danio rerio). Different concentrations of PEAOXA were tested for toxicity on zebrafish embryonic development; only the highest concentration of 30 mg/L showed toxic effects. Quantitative RT-PCR was applied to detect Tumor necrosis factor-α, Interleukin 1β, 6, and 8, and members of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB). Exposure to LPS induced an increase in pro-inflammatory cytokines (tumor necrosis factor and interleukin 1, 6, and 8) in both gene and protein expression, as well as an increase of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) and the nuclear factor kappa light polypeptide enhancer in B-cells inhibitor (IκBα) gene expression. Furthermore, acute LPS exposure also induced an increase in tryptase release, related to mast cell activity, and in the production of apoptosis-related proteins (caspase 3, bax, and bcl-2). Treatment with PEAOXA 10 mg/L significantly counteracts LPS-induced inflammation in terms of cytokine expression and decreases tryptase release and the apoptosis pathway.

Environmental pollutants may cause adverse effects on the immune system of aquatic organisms. This study revealed that combination of environmental pollutants and Bisphenol A(BPA) could cause an acute inflammatory response in zebrafish larvae as shown by body alterations, which may imply a common immunotoxicity mechanism for most environmental pollutants. In the present study we evaluated the toxicity after co-exposure of BPA and Cd or Cr (III) in zebrafish embryos and larvae, and the oxidative stress pathway involved. Evaluation of lethal and developmental endpoints such as hatching, edema, malformations, abnormal heart rate and survival rate were evaluated after 96 h of exposure. Combination of BPA at 10 μM with Cd or Cr at 0.5 μM exposure induce malformations at 96 hpf in zebrafish larvae, as well as significantly increases oxidative stress and induce apoptosis on larvae. Our study suggested how environmental pollutant showed a synergistic effect at common not-effective doses, promoting decrease of antioxidant defense and contrasted fish development.

Aflatoxin B1 (AFB1), a secondary metabolite produced by fungi of the genus Aspergillus, has been found among various foods as well as in fish feed. However, the effects of AFB1 on fish development and its associated toxic mechanism are still unclear. In the present study, we confirmed the morphological alterations in zebrafish embryos and larvae after exposure to different AFB1 doses as well as the oxidative stress pathway that is involved. Furthermore, we evaluated the potentially protective effect of Hericium erinaceus extract, one of the most characterized fungal extracts, with a focus on the nervous system. Treating the embryos 6 h post fertilization (hpf) with AFB1 at 50 and 100 ng/mL significantly increased oxidative stress and induced malformations in six-day post-fertilization (dpf) zebrafish larvae. The evaluation of lethal and developmental endpoints such as hatching, edema, malformations, abnormal heart rate, and survival rate were evaluated after 96 h of exposure. Hericium inhibited the morphological alterations of the larvae as well as the increase in oxidative stress and lipid peroxidation. In conclusion: our study suggests that a natural extract such as Hericium may play a partial role in promoting antioxidant defense systems and may contrast lipid peroxidation in fish development by counteracting the AFB1 toxicity mechanism.

Data on the biology and ecology of Galeus melastomus are old/absent for the Southern Tyrrhenian Sea, despite there being numerous studies in the wider area. A total of 127 specimens of G. melastomus from the southern Tyrrhenian Sea, collected in 2018–2019 using trawling nets, were analyzed to investigate size at sexual maturity, sex ratio, length–weight relationships, and feeding habits. To our best knowledge, this is the first time in which all these features were investigated in the Southern Tyrrhenian Sea for G. melastomus. The stomach content analysis showed that G. melastomus had intermediate feeding habits, preying on a great variety of species, especially Cephalopoda, Osteichthyes, and Crustacea. The Levin’s index value (Bi) was 0.53. Sex ratio was 0.92:1, with females slightly more abundant and bigger than males. The results also showed a decrease (33.7 cm for females, 31.1 cm for males) in length at 50% maturity (L50). This could be a result of anthropogenic stressors, such as overfishing and/or and environmental changes, which can induce physiological responses in several species. Our results highlighted the differences related to sexual maturity, growth, and feeding habits of the blackmouth catshark in the studied area, providing reference data to allow comparison with future studies on this species adaptations to this and other deep-sea areas in the Mediterranean Sea.

Beachrock can be found in intertidal marine environments worldwide even though the knowledge about its ecological value in Messina's strait is still quite scarce. In this study, six tidal ponds of Messina's Beachrock were sampled from October 2016 to October 2017 to investigate the ecological status of the area, biotic and abiotic factors that can influence the faunal assemblage and the relationships among the communities inhabiting the various pools examined. A total of 13399 organisms, belonging to 5 Phyla, were identified by Visual Census. The surveyed stations showed biodiversity values and evenness that depend, above all, from the edaphic features and water exchange of the different tidal ponds. Tidal ponds characterized by a lower water exchange showed a faunal assemblage characterized by the presence of species tolerant to a wide temperature and salinity range, as Mugil cephalus, Clibanarius erythropus and Phorcus turbinatus. The pools with the most water exchange showed the highest abundance and evenness values; while the ponds characterized by hard bottom showed this feature as strong influence on the species composition. A preliminary mathematical study was also carry out to describe, qualitatively, the main mechanisms ruling the interaction between the Clibanarius erythropus and Phorcus turbinatus.

In the present study, it has been evaluated how 10 µm of polyethylene microspheres can be ingested by Artemia salina (Linnaeus, 1758) larvae within the first 7 days of the life cycle, and the impact on their health. Twelve A. salina larvae (instar I) groups were exposed to different microplastics (MPs) concentrations (0-1-10-102-103-104 MPs/mL), with and without Dunaliella salina as a food source. The results highlighted that A. salina larvae ingest MPs in relation to the exposure times in a dose-dependent manner and are significantly influenced by food availability. The highest contamination found was 306.2 MPs/individual at 104 MPs/mL exposure without a food source. No MPs were found in the presence of the food source from 1 to 102 MPs/mL, while contamination was detected at all concentrations of MPs without a food source. The worst effect on the developmental stages was evaluated at 168 h with a food source, with a delay compared to the control of I and II instars at 103 and 104 MPs/mL, respectively. Furthermore, microalgal feeding was significantly reduced for about 50% in the presence of 104 MPs/mL. These results highlight that aquatic microplastics pollution could affect the A. salina’s feeding behavior and life cycle.

Cnidarian toxins represent a rich source of biologically active compounds. Since they may act via oxidative stress events, the aim of the present study was to verify whether crude venom, extracted from the jellyfish Pelagia noctiluca, elicits inflammation and oxidative stress processes, known to be mediated by Reactive Oxygen Species (ROS) production, in rats. In a first set of experiments, the animals were injected with crude venom (at three different doses 6, 30 and 60 µg/kg, suspended in saline solution, i.v.) to test the mortality and possible blood pressure changes. In a second set of experiments, to confirm that Pelagia noctiluca crude venom enhances ROS formation and may contribute to the pathophysiology of inflammation, crude venom-injected animals (30 µg/kg) were also treated with tempol, a powerful antioxidant (100 mg/kg i.p., 30 and 60 min after crude venom). Administration of tempol after crude venom challenge, caused a significant reduction of each parameter related to inflammation. The potential effect of Pelagia noctiluca crude venom in the systemic inflammation process has been here demonstrated, adding novel information about its biological activity.

The phylum Cnidaria is one of the most important contributors in providing abundance of bio- and chemodiversity. In this study, a comprehensive chemical investigation on the nutritional and nutraceutical properties of Mediterranean jellyfish Pelagia noctiluca was carried out. Also, compositional differences between male and female organisms, as well as between their main anatomical parts, namely bell and oral arms, were explored in an attempt to select the best potential sources of nutrients and/or nutraceuticals from jellyfish. With the exception of higher energy densities and total phenolic contents observed in females than males, no statistically significant differences related to the specimen’s sex were highlighted for the other compound classes. Rather, the distribution of the investigated chemical classes varied depending on the jellyfish’s body parts. In fact, crude proteins were more abundant in oral arms than bells; saturated fatty acids were more concentrated in bells than oral arms, whereas polyunsaturated fatty acids were distributed in the exact opposite way. On the other hand, major elements and trace elements demonstrated an opposite behavior, being the latter most accumulated in oral arms than bells. Additionally, important nutraceuticals, such as eicosapentaenoic and docosahexaenoic acids, and antioxidant minerals, were determined. Overall, obtained data suggest the potential employment of the Mediterranean P. noctiluca for the development of natural aquafeed and food supplements.

Enormous marine biodiversity offers an endless reservoir of chemicals for many applications. In this scenario, the extraction of seaweeds represents an interesting source of compounds displaying antimicrobial activity. In particular, among the different red algae, Gracilaria gracilis plays an important role due to the presence of important bioactives in its composition. In spite of these features, an efficient culture system is still absent. In the present study, a novel algal culture method was developed and compared to another more common cultural practice, widely reported in literature. A higher efficiency of the new method, both for daily growth rate and biomass, was assessed. Furthermore, the growth inhibitory activity of five extracts, obtained using ethanol, methanol, acetone, chloroform or diethyl ether as a solvent, from the cultured G. gracilis was tested against Gram-positive and Gram-negative pathogens. Algal extracts exhibited a considerable inhibitory activity against B. subtilis strains, while a slight inhibition was observed against V. fischeri. The different extracts showed significant differences in bacterial growth inhibition, with the highest activity that was recorded for the ethanol extract, followed by that of methanol. Based on the chemical characterization, these findings could be related to the antimicrobial activity played by the combination of total carbohydrates and polyphenols, which were determined at high levels in ethanol and methanol extracts, as well as by the highest number and levels of single polyphenols. Conversely, the lower growth inhibitory activities found in chloroform and diethyl ether extracts could be related to the isolation of minor lipid classes (e.g., neutral and medium polar lipids) composed by fatty acids, such as stearic, oleic and arachidonic acids, typically characterized by antimicrobial activity. In consideration of the results obtained, the present study has a double implication, involving both the field of cultural practices and the exploitation of natural sources for the isolation of antimicrobial agents useful both in pharmaceutical and food applications.

The biological activity of extract obtained from the ascidians Styela plicata and Ascidia mentula, living in the coastal lake Faro (Messina, Italy) was ascertained with specific regard to cytotoxic effect, tested on cultured human embryonic kidney (HEK 293 Phoenix) cells after 12, 24 or 48 h, and to antimicrobial effect (10 mg/mL), assayed on both Gram positive and negative strains. Nitrosative stress, possibly induced by the extract on HEK 293 cells and assessed by NO2 –/NO3 – production measurement, was also verified. With regard to cytotoxic activity, A. mentula extract was more effective than S. plicata one, as shown by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, with toxic action due to nitrosative stress. S. plicata extract had a more significant antimicrobial activity than A. mentula. The present findings provide a first evidence of the biological power exhibited by both S. plicata and A. mentula extracts, thus increasing the knowledge about the biological activity of marine compounds and providing novel information to possibly correlate the different toxicity pattern displayed by both specimens and their distribution in the lake Faro (Messina).

ABSTRACT In this research, the seasonal fluctuations of the main water variables in the no-take area (A zone) of the Ustica MPA were investigated. This study aims to monitor the water quality of the Ustica MPA, to analyze all the hydrological parameters useful to the physical-chemical characterization of water and to use TRIX index to characterize the trophic level of the coastal marine area. The final results shown that Ustica is an area of relevant environmental and ecological value and is a good example of a Marine Protected Area, which is not excessively affected by the human presence even during the touristic season. This study represents the first records based on well-organized protocol in order to assess water column characteristics of the studied area. This paper could be used as a reference document for future studies regarding the same or similar areas.