Food from the Depths of the Mediterranean: The Role of Habitats, Changes in the Sea-Bottom Temperature and Fishing Pressure.

As part of the "Innovations in the Food System: Exploring the Future of Food" Special Issue, this paper briefly reviews studies that highlight a link between deep-sea fishery resources (deep-sea food resources) and vulnerable marine ecosystems (VME), species, and habitats in the Mediterranean Sea, providing new insights into changes in commercial and experimental catches of the deep-sea fishery resources in the central Mediterranean over the last 30 years. About 40% of the total landing of Mediterranean deep-water species is caught in the central basin. Significant changes in the abundance of some of these resources with time, sea-bottom temperature (SBT), and fishing effort (FE) have been detected, as well as an effect of the Santa Maria di Leuca cold-water coral province on the abundance of the deep-sea commercial crustaceans and fishes. The implications of these findings and the presence of several geomorphological features, sensitive habitats, and VMEs in the central Mediterranean are discussed with respect to the objectives of biodiversity conservation combined with those of management of fishery resources.

Use of Biochar to Improve the Sustainable Crop Production of Cauliflower (Brassica oleracea L.).

In agriculture, biochar (B) application has been suggested as a green technology to reduce nitrate pollution from agricultural origins and improve crop yield. The agronomic impact of B use on soil has been extensively studied, while knowledge of its possible effects on horticultural cultivation is still scarce. A greenhouse experiment was conducted to evaluate the effect of using biochar in soils treated with two different rates of nitrogen fertilizers on soil properties and nitrogen (N) leachate. This study also investigated the vegetative parameters during the crop growing season of Brassica oleracea L. var. botrytis. Soil mesocosms were set up to test the following treatments: untreated/control (C); normal dose of N fertilizer (130 kg N ha-1) (ND); ND+B; high dose of N fertilizer (260 kg N ha-1) (HD); and HD+B. Principal component analysis and cluster analysis were exploited to assess biochar's ability to reduce nitrate leaching and enhance soil-vegetative properties. Biochar addition affected the soil chemical properties of the fertilized microcosms (ND and HD). Biochar increased the NH4+ content in HD soil and the NO3- content in ND soil by 26 mg/L and 48.76 mg/L, respectively. The results showed that biochar application increased the marketable cauliflower yield. In ND+B and HD+B, the curd weight was 880.68 kg and 1097.60 kg, respectively. In addition, a small number of nitrogenous compounds in the leachate were quantified in experimental lines with the biochar. Therefore, biochar use improves the marketable yield of horticulture, mitigating the negative impacts associated with the mass use of N fertilizers in agriculture.

Occurrence and spatial distribution of dioxin and dioxin-like compounds in topsoil of Taranto (Apulia, Italy) by GC-MS analysis and DR-CALUX® bioassay.

The aim of the present study was to assess the occurrence and spatial distribution of PCDD/Fs and dioxin-like compounds in topsoils of Taranto (Apulia Region), one of the most heavily industrialized and contaminated area of Southern Italy. A combined approach of chemical analysis by GC-MS/MS and AhR reporter gene bioassay was applied in a subset of topsoil samples (n = 20) collected in 2017-18 from ten sites embracing three levels of risk (from high to low) in the framework of a large survey inside Taranto municipality. TCDD-BEQs and GC-MS/MS TEQWHO and TEQTHEORETICAL revealed a decreasing trend with the distance from main industrial settings and landfill areas. A strong correlation between TCDD-BEQs and TEQWHO values (R2 = 0.85) and TEQTHEORETICAL (R2 = 0.88) was also found. In 3 out of 10 topsoil investigated, BEQs and TEQWHO/THEORETICAL resulted above Italian National Regulatory Limits for ∑PCDD/Fs in green, private and recreational used soils (10 ng TEQ/kg d.w. D.Lgs 152/2006) and for ∑PCDD/F/dl-PCBs in agricultural and farming soil (6 ng TEQ/kg d.w. D.M. 46/2019). GC-MS/MS pattern revealed the highest prevalence of dl-PCBs in 6 out of 10 sites, followed by PCDFs and PCDDs. Those sites are all located in proximity of main industrial steel and iron ore sinter plant, steel plant's landfills and illegal dumping sites. An update on occurrence and spatial distribution of PCDD/Fs and dl-PCBs contamination of Taranto urban soils was obtained and the DR-CALUX® bioassay was further recommended as a suitable screening tool for environmental and human risk assessment.

First Attempt to Couple Proteomics with the AhR Reporter Gene Bioassay in Soil Pollution Monitoring and Assessment.

A topsoil sample obtained from a highly industrialized area (Taranto, Italy) was tested on the DR-CALUX® cell line and the exposed cells processed with proteomic and bioinformatics analyses. The presence of polyhalogenated compounds in the topsoil extracts was confirmed by GC-MS/MS analysis. Proteomic analysis of the cells exposed to the topsoil extracts identified 43 differential proteins. Enrichment analysis highlighted biological processes, such as the cellular response to a chemical stimulus, stress, and inorganic substances; regulation of translation; regulation of apoptotic process; and the response to organonitrogen compounds in light of particular drugs and compounds, extrapolated by bioinformatics all linked to the identified protein modifications. Our results confirm and reflect the complex epidemiological situation occurring among Taranto inhabitants and underline the need to further investigate the presence and sources of inferred chemicals in soils. The combination of bioassays and proteomics reveals a more complex scenario of chemicals able to affect cellular pathways and leading to toxicities rather than those identified by only bioassays and related chemical analysis. This combined approach turns out to be a promising tool for soil risk assessment and deserves further investigation and developments for soil monitoring and risk assessment.

Ecosystem functioning of two marine food webs in the North-Western Ionian Sea (Central Mediterranean Sea).

The ecosystem functioning of two marine food webs covering the north-eastern (Salento) and south-western (Calabria) sectors of the North-Western Ionian Sea (NWIS) (Central Mediterranean Sea) was investigated through a food-web model. Data inputs covered a wide set of ecological information applied to 58 functional groups (FGs). The sum of consumption and the mean predation mortality rate were calculated for benthic, demersal, and pelagic subsystems indicating the predator and prey roles of the FGs. A complex system of energy and biomass exchanges characterized the investigated food webs indicating an important benthic-pelagic coupling. In the food webs of both areas, the regulation of flows between the benthic-pelagic coupling seems to occur through the benthopelagic shrimps and the small pelagics due to their wasp-waist control role. Differences were observed concerning the top predators. Odontocetes play this keystone role in the Salento food web. Anglers, bathyal squids, and sharks assume this functional role in Calabria. The geomorphology and hydrography in the NWIS could affect the biomass and energy exchanges in this coupling. The higher flows of consumption of the benthic system observed in the Calabria food web could be influenced by a widespread presence of canyons along the continental edge which increase the benthic productivity. In contrast, the flows of consumption in the Salento food web seem to be driven by the planktonic productivity supporting the pelagic, benthopelagic, and demersal compartments. This condition could be favored by the large extension of the shelf break zone. The food-web models realized for the NWIS represent ideal platforms for the development of analysis with dynamic simulations. The comparative analysis of the two food webs by means of the FGs and their functional traits allowed the general pattern of ecosystem structure and functioning in the NWIS to be identified, making it an interesting approach to investigate the marine ecosystem.

Towards Non-Invasive Methods to Assess Population Structure and Biomass in Vulnerable Sea Pen Fields.

Colonies of the endangered red sea pen Pennatula rubra (Cnidaria: Pennatulacea) sampled by trawling in the northwestern Mediterranean Sea were analyzed. Biometric parameters, such as total length, peduncle length, number of polyp leaves, fresh weight, and dry weight, were measured and related to each other by means of regression analysis. Ad hoc models for future inferencing of colonies size and biomass through visual techniques were individuated in order to allow a non-invasive study of the population structure and dynamics of P. rubra.

Haplosporidium pinnae associated with mass mortality in endangered Pinna nobilis (Linnaeus 1758) fan mussels.

The fan mussel, Pinna nobilis (Linnaeus 1758), is an endemic bivalve of the Mediterranean basin, protected by international legislation as an endangered species. In the early summer of 2018, a mass mortality event (MME) of P. nobilis was recorded in the Gulf of Taranto (Southern Italy, Ionian Sea). Moribund specimens of P. nobilis were collected by scuba divers and processed by bacteriological, parasitological, histopathological and molecular analyses to investigate the causes of this MME. Different developmental stages (i.e., plasmodia, spores and sporocysts) of a presumptive haplosporidian parasite were observed during the histological analysis in the epithelium and in the lumen of the digestive tubules, where mature spores occurred either free or in sporocysts. The spores presented an operculum and an ovoid shape measuring 4.4 µm (±0.232) in length and 3.6 µm (±0.233) in width. BLAST analysis of an 18SrRNA sequence revealed a high nucleotide similarity (99%) with the reference sequence of Haplosporidium pinnae available in GenBank database. Phylogenetic analysis clustered the sequence of the pathogen in a paraphyletic clade with the reference sequence of H. pinnae, excluding other haplosporidians (i.e., Bonamia and Minchinia genera). Based on data reported, H. pinnae was the causative agent of MME in the populations of P. nobilis sampled in the Ionian Sea, where the conservation of this endangered species is heavily threatened by such a protozoan infection. Further investigations should contribute to knowledge about the life cycle of H. pinnae in order to reduce spread of the pathogen and to mitigate the burden of the disease where P. nobilis is facing the risk of extinction.

Improving the Conservation of Mediterranean Chondrichthyans: The ELASMOMED DNA Barcode Reference Library.

Cartilaginous fish are particularly vulnerable to anthropogenic stressors and environmental change because of their K-selected reproductive strategy. Accurate data from scientific surveys and landings are essential to assess conservation status and to develop robust protection and management plans. Currently available data are often incomplete or incorrect as a result of inaccurate species identifications, due to a high level of morphological stasis, especially among closely related taxa. Moreover, several diagnostic characters clearly visible in adult specimens are less evident in juveniles. Here we present results generated by the ELASMOMED Consortium, a regional network aiming to sample and DNA-barcode the Mediterranean Chondrichthyans with the ultimate goal to provide a comprehensive DNA barcode reference library. This library will support and improve the molecular taxonomy of this group and the effectiveness of management and conservation measures. We successfully barcoded 882 individuals belonging to 42 species (17 sharks, 24 batoids and one chimaera), including four endemic and several threatened ones. Morphological misidentifications were found across most orders, further confirming the need for a comprehensive DNA barcoding library as a valuable tool for the reliable identification of specimens in support of taxonomist who are reviewing current identification keys. Despite low intraspecific variation among their barcode sequences and reduced samples size, five species showed preliminary evidence of phylogeographic structure. Overall, the ELASMOMED initiative further emphasizes the key role accurate DNA barcoding libraries play in establishing reliable diagnostic species specific features in otherwise taxonomically problematic groups for biodiversity management and conservation actions.

Environmental hazard of yperite released at sea: sublethal toxic effects on fish.

The aim of this study was to evaluate the potential toxicological effects on fish related to the leakage of yperite from rusted bomb shells dumped at sea. Both in vivo and field studies have been performed. As for the in vivo experiment, specimen of European eel were subcutaneously injected with 0.015, 0.15 and 1.5mg/kg of yperite and sacrificed after 24 and 48 h. In the field study, specimen of Conger eel were collected from a dumping site in the Southern Adriatic Sea. The presence/absence of yperite in tissues, genotoxicity, detoxification enzymes, histological alterations and gross abnormalities were investigated. Results of the in vivo experiment showed a significant increase of EROD activity at both 24h and 48 h. UGT activity increased significantly at 48 h post injection. An acute inflammatory response after 24h in skin layers and muscle was observed, associated to cell degeneration and necrosis after 48 h at the highest dose. On field, comet assay revealed genotoxicity in gills of fish from the dumping site. Specimen from the dumping site showed significantly higher EROD activities compared to controls, deep ulcers and papules on skin together with liver and spleen histopathological lesions.

Comparing deep-sea fish fauna between coral and non-coral "megahabitats" in the Santa Maria di Leuca cold-water coral province (Mediterranean Sea).

Two experimental longline surveys were carried out in the Santa Maria di Leuca (SML) cold-water coral province (Mediterranean Sea) during May-June and September-October 2010 to investigate the effect of corals on fish assemblages. Two types of "megahabitat" characterized by the virtual absence of fishing were explored. One was characterized by complex topography including mesohabitats with carbonate mounds and corals. The other type of megahabitat, although characterized by complex topographic features, lacks carbonate mounds and corals. The fishing vessel was equipped with a 3,000 m monofilament longline with 500 hooks and snoods of 2.5 m in length. A total of 9 hauls, using about 4,500 hooks, were carried out both in the coral megahabitat and in the non-coral megahabitat during each survey. The fish Leucoraja fullonica and Pteroplatytrygon violacea represent new records for the SML coral province. The coral by-catch was only obtained in the coral megahabitat in about 55% of the stations investigated in both surveys. The total catches and the abundance indices of several species were comparable between the two habitat typologies. The species contributing most to the dissimilarity between the two megahabitat fish assemblages were Pagellus bogaraveo, Galeus melastomus, Etmopterus spinax and Helicolenus dactylopterus for density and P. bogaraveo, Conger conger, Polyprion americanus and G. melastomus for biomass. P. bogaraveo was exclusively collected in the coral megahabitat, whereas C. conger, H. dactylopterus and P. americanus were found with greater abundance in the coral than in the non-coral megahabitat. Differences in the sizes between the two megahabitats were detected in E. spinax, G. melastomus, C. conger and H. dactylopterus. Although these differences most probably related to the presence-absence of corals, both megahabitats investigated play the role of attraction-refuge for deep-sea fish fauna, confirming the important role of the whole SML coral province as a refuge area from fishing.

DNA damage, severe organ lesions and high muscle levels of As and Hg in two benthic fish species from a chemical warfare agent dumping site in the Mediterranean Sea.

The aim of the present study was to evaluate the environmental threat to benthic species from chemical weapons dumped in the southern Adriatic Sea. An ecotoxicological approach using chemical analysis and biological responses was applied, in two sentinel species: the Blackbelly rosefish Helicolenus dactylopterus and European conger Conger conger. Specimen were collected in a stretch of sea, where had been dumped war materials and from a reference site free of ordnance. Residues of yperite, Hg and As were measured in fish fillets. Skin, liver, kidney and spleen were examined for histopathological and macroscopical lesions. Liver detoxifying capacities (EROD and UDPGT) and genotoxicity (comet assay) were also investigated. As and Hg levels were three-four times higher than those from the reference site in both species (p<0.001). Both species captured in dumping site showed clear signs of chronic illness according to the health assessment index (HAI). Deep ulcers and nodules were observed on skin and external organs. Histological lesions such as periportal and bile duct fibrosis, pericholangitis, steatosis, granuloma and elevated splenic MMCs were detected in liver and spleen. Significantly higher EROD activities were also found in both species from dumping site (p<0.01). Comet assay revealed genotoxicty in gills of C. conger from dumping site, indicating uptake of chemical warfare agents through fish gills. European conger was found to be a more sensitive bioindicator of this type of contamination than the Blackbelly rosefish.