Marine macro-litter mass outweighs biomass in trawl catches along abyssal seafloors of Sardinia channel (Italy).

This study provides new insights onto spatial and temporal trends of seafloor macro-litter in the abyssal seafloor of Sardinian channel, in central western Mediterranean (Italy). Trawl surveys were conducted at depths between 884 and 1528 m, thus focusing on one of the least investigated marine environments. None of the considered sites was litter free, with plastics being numerically dominant (57% of items), followed by metal (11%) and glass (16%). Recorded densities and weight ranged between 49.9 and 499 items km-2 and 1.4 and 1052 kg km-2. In the most contaminated sites, the weight of the litter collected in nets represented up to nine times the biomass of benthic megafauna, and, overall, in 60% of hauls macro-litter mass outweighed the biomass collected. Moreover, we report that megafauna was observed to be more abundant in sites where macro-litter presence was more severe. More studies are needed to elucidate the nature of this correlation, with biota being more abundant in hotspots of accumulation of seafloor macro-litter.

I Like This New Me: Unravelling Population Structure of Mediterranean Electric Rays and Taxonomic Uncertainties within Torpediniformes.

The present study focused on the three species of electric rays known to occur in the Mediterranean Sea: Torpedo torpedo, Torpedo marmorata and Tetronarce nobiliana. Correct identification of specimens is needed to properly assess the impact of fisheries on populations and species. Unfortunately, torpedoes share high morphological similarities, boosting episodes of field misidentification. In this context, genetic data was used (1) to identify specimens caught during fishing operations, (2) to measure the diversity among and within these species, and (3) to shed light on the possible occurrence of additional hidden species in the investigated area. New and already published sequences of COI and NADH2 mitochondrial genes were analyzed, both at a small scale along the Sardinian coasts (Western Mediterranean) and at a large scale in the whole Mediterranean Sea. High levels of genetic diversity were found in Sardinian populations, being significantly different from other areas of the Eastern Mediterranean Sea due to the biotic and abiotic factors here discussed. Sardinian torpedoes can hence be indicated as priority populations/areas to be protected within the Mediterranean Sea. Moreover, sequence data confirmed that only the three species occur in the investigated area. The application of several 'species-delimitation' methods found evidence of cryptic species in the three species outside the Mediterranean Sea, as well as in other genera/families, suggesting the urgent need for future studies and a comprehensive revision of the order Torpediniformes for its effective conservation.

Ontogenetic Shifts in Body Morphology of Demersal Sharks' Species (Order: Squaliformes) Inhabiting the Western-Central Mediterranean Sea, with Implications for Their Bio-Ecological Role.

Several elasmobranch species undergo shifts in body proportions during their ontogenetic growth. Such morphological changes could reflect variation in diet, locomotion, or, more broadly, in the species' interactions with their environment. However, to date, only a few studies have been conducted on this topic, and most of them focused on particular body regions. In the present study, the ontogenetic growth of five different demersal shark species was investigated by using both traditional linear morphometry of the entire body and shape analysis of the caudal fin. A total of 449 sharks were analysed: 95 little gulper sharks, 80 longnose spurdogs, 103 kitefin sharks, 124 velvet belly lanternsharks, and 47 angular roughsharks. From each specimen, 36 linear morphometric measurements were taken. While a first canonical analysis of principal coordinates ruled out the possibility of different growth patterns between males and females, the same analysis statistically discriminated between small and large individuals in every species based on their morphology. A Similarity Percentage analysis revealed that the most important measurements in distinguishing these two groups were those related to body lengths, indicating that large individuals are more elongated than small individuals. The shape analysis of caudal fins revealed allometric growth during ontogenetic development, with adult individuals having a wider fin (discriminant analysis, p < 0.05). These findings could be related to changes in predatory skills, supporting the hypothesis of a shift in the ecological role that these sharks play in their environment, thus providing new essential information for their conservation.

Movement patterns analysis as a tool in Fully Protected Areas design: Influence of relocations on travelled distances of Palinurus elephas (Fabr. 1787) in Sardinian FPAs (central-western Mediterranean).

Effectiveness of restocking programs in Marine Fully Protected Areas' (FPAs) can be highly affected by the movement behavior of the species to protect. We analyzed the data of 744 Palinurus elephas specimens tagged, relocated inside 12 FPAs, established in the seas surrounding Sardinia (central-western Mediterranean), and then recaptured. The overall aims were to characterize the individuals' movements after the relocation and to discuss strengths and weaknesses of the current FPAs' design. Almost 80% of tagged individuals travelled progressively increasing distances, as a function of the release distance, up to a maximum value of 11,500 m. Then, for larger release distances, individuals' mobility appeared reduced and more chaotic. A similar trend was found analyzing the distances travelled with respect to the FPAs with the highest number of recaptured individuals and to the size. These results allow to establish a threshold limit of the release distance, to take into account when designing FPAs, as a useful tool to keep a portion of the individuals into FPAs' borders, in order to both preserve the species and guarantee a spillover of individuals in the commercial areas.

The Use of Reproductive Indicators for Conservation Purposes: The Case Study of Palinurus elephas in Two Fully Protected Areas and Their Surrounding Zones (Central-Western Mediterranean).

In 1990s, the European spiny lobster Palinurus elephas, one of the most commercially important species in the Mediterranean, exhibited a population decline. For this reason, fully protected areas (FPAs) appeared effective in re-establishing natural populations and supporting fishery-management objectives. Here, the reproductive parameters of P. elephas populations in two different FPAs (Su Pallosu and Buggerru, central-western Mediterranean), where a restocking programme was carried out, and in their surrounding commercial zones, were investigated from quantitative and qualitative perspectives. The comparison of fecundity between females collected inside and outside FPAs did not show statistical differences as well as the vitellogenin concentration, which did not vary among eggs of different size classes of females caught inside and outside the FPAs, indicating the same reproductive potential. The study demonstrated a benefit of overexploited populations in terms of enhancement of egg production overtime (15 years for Su Pallosu and 6 years for Buggerru) with a mean egg production 4.25-5.5 times higher at the end of the study than that observed at the beginning of the study. The main driver of eggs production appeared to be size, with larger lobsters more present inside the FPAs than outside. Given these results, the dominant contribution of the two studied FPAs to the regional lobster reproduction is remarkable.

Investigating the Ovarian Microstructure in the Genera Helicolenus and Scorpaena (Teleostei, Sub-Order Scorpaenoidei) with Implications for Ovarian Dynamics and Spawning.

The sub-order Scorpenoidei appears to be particularly interesting due to the presence of intermediate stages between oviparity and viviparity in several species. The present study aims to describe the ovarian morphology, using a histological and histochemical approach, in four ovuliparous species belonging to Scorpaena genus compared with a zygoparous species, H. dactylopterus, focusing also on the assessment of the ovarian dynamics in the populations of such species in Sardinia waters (central-western Mediterranean). Ovarian sections of all species were examined using light microscopy. All species showed a specialized ovary, cystovarian type II-3, strictly related to the production of gelatinous matrices surrounding the eggs. Some microscopic peculiarities in the oogenesis process were found: thin zona pellucida, small and low cortical alveoli, and a specialized ovarian wall during the spawning period. All species analyzed were batch-spawners with an asynchronous ovarian organization. A continuous recruitment of oocytes and the occurrence of de novo vitellogenesis was also observed. During the spawning period, low atresia intensity was detected, while a marked increase in this intensity found in the ovaries at the end of spawning season. Our observations may support an indeterminate fecundity type for these species.

Scattered accumulation hotspots of macro-litter on the seafloor: Insights for mitigation actions.

Marine litter is an ever-increasing problem that demands immediate reduction plans and mitigation actions that should act synergically to efficiently meet ambitious goals. Since the seafloor has been recognized as the major sink for marine debris, the study of litter accumulation dynamics represent a fundamental tool to evaluate possible removal actions. We analysed a 7 years (2013-2019) standardized data series collected along Sardinian fishing grounds through MEDiterranean International Trawl Survey, for which estimates of density and weight of seafloor macro-litter were calculated for over 707 hauls. Results show the absence of any temporal trend in seafloor macro-litter density and weight, but rather indicate a spatial and bathymetric segregation of different litter categories. Our data showed how different sources and physical features of macro-litter items (i.e., plastic and fishing gear, rubber, glass, metals, and cloth) led to spatially segregated accumulation hotspots. We also point out here how the identification of seafloor macro-litter hotspots using aggregated data that include plastic items could obscure the identification of other segregated but yet relevant hotspots of other macro-litter categories accumulated in the marine environment. These hotspots often occurred at shallower depths and closer to coastlines, thus representing potential spots where eventual future litter removal action could be prioritized.

A Taxonomic Survey of Female Oviducal Glands in Chondrichthyes: A Comparative Overview of Microanatomy in the Two Reproductive Modes.

Oviducal glands (OGs) are distinct expanded regions of the anterior portion of the oviduct, commonly found in chondrichthyans, which play a key role in the production of the egg in-vestments and in the female sperm storage (FSS). The FSS phenomenon has implications for understanding the reproductive ecology and management of exploited populations, but little information is available on its taxonomic extent. For the first time, mature OGs from three lecithotrophic oviparous and four yolk-sac viviparous species, all considered at risk from the fishing impacts in the central western Mediterranean Sea, were examined using light microscopy. The OG microanatomy, whose morphology is generally conserved in all species, shows differences within the two reproductive modalities. Oviparous species show a more developed baffle zone in respect to viviparous ones because of the production of different egg envelopes produced. Among oviparous species, Raja polystigma and Chimaera monstrosa show presence of sperm, but not sperm storage as observed, instead, in Galeus melastomus and in all the viviparous sharks, which preserve sperm inside of specialized structures in the terminal zone.

Bio-Ecological Features Update on Eleven Rare Cartilaginous Fish in the Central-Western Mediterranean Sea as a Contribution for Their Conservation.

Cartilaginous fish are commonly recognized as key species in marine ecosystems for their fundamental ecological role as top predators. Nevertheless, effective management plans for cartilaginous fish are still missing, due to the lack of knowledge on their abundance, distribution or even life-history. In this regard, this paper aims at providing new information on the life-history traits, such as age, maturity, reproductive period, in addition to diet characteristics of eleven rare cartilaginous fish inhabiting the Central-Western Mediterranean Sea belonging to the orders Chimaeriformes (Chimaera monstrosa), Hexanchiformes (Heptranchias perlo and Hexanchus griseus), Myliobatiformes (Aetomylaeus bovinus and Myliobatis aquila), Rajiformes (Dipturus nidarosiensis and Leucoraja circularis), Squaliformes (Centrophorus uyato, Dalatias licha and Oxynotus centrina) and Torpediniformes (Tetronarce nobiliana), useful for their assessment and for future management actions. Particularly, the present paper provides for the first time the age estimation of D. nidarosienis and L. circularis which were both found capable of becoming older than ten years. In addition, the present study updates the sizes of first maturity of C. uyato and D. licha, which appeared to be capable of reproducing earlier than what was previously hypothesized, representing very valuable information for a better understanding of these rare species populations status and, eventually, their conservation. On the basis of the stomach content analysis, it was possible to identify five different predator groups.

Scattered accumulation hotspots of macro-litter on the seafloor: Insights for mitigation actions.

Marine litter is an ever-increasing problem that demands immediate reduction plans and mitigation actions that should act synergically to efficiently meet ambitious goals. Since the seafloor has been recognized as the major sink for marine debris, the study of litter accumulation dynamics represents a fundamental tool to evaluate future removal actions. We analysed a 7 years (2013-2019) standardized data series collected along Sardinian fishing grounds through MEDiterranean International Trawl Survey, for which estimates of density and weight of seafloor macro-litter were calculated over 707 hauls. Results show the absence of any temporal trend in seafloor macro-litter density and weight, but rather indicate a spatial and bathymetric segregation of different litter categories. Our data showed how different sources and physical features of macro-litter items (i.e., plastic and fishing gears, rubber, glass, metal and textile) led to spatially segregated accumulation hotspots. These hotspots often occurred at shallower depths and closer to coastlines, representing spots where future litter removal action could be prioritized. We also point out here how the identification of seafloor macro-litter hotspots using aggregated data that include plastic items could indeed hide the identification of hotspots of other less abundant but yet detrimental macro-litter categories accumulated in the marine environment.

New insights into life-history traits of Mediterranean Electric rays (Torpediniformes: Torpedinidae) as a contribution to their conservation.

Despite being commonly recognized by the general public, knowledge about the biology and life-history of electric rays is still scarce. Globally, according to the International Union for Conservation of Nature, more of the 80% of the species belonging to the Torpedinidae family are, indeed, classified as Data Deficient, so as, in most of the cases, no proper management measures are available for the conservation of these species. For this reason, the main goal of this study is to provide new information on life-history traits of three species inhabiting the Mediterranean Sea. Age and growth were estimated through direct annuli count of 146 vertebral centra sections of Torpedo marmorata. Females' growth pattern was found to be better described by a S-shaped function (Gompertz L∞ = 622mm TL; k = 0.15; IP=2.75 years), while males' by a logarithmic one (von Bertalanffy L∞ = 485mm TL; k = 0.14; t0 = -2.28). The oldest female and male were aged 17 (560mm TL) and 11 (432mm TL), respectively. Females (L50=379mm TL and A50=5.5 years) appeared to mature later than males (L50=272mm TL and A50=3.8 years), also attaining bigger sizes. Despite the observation of late-pregnant females being found only in autumn and regressing ones in autumn and winter, the presence of juveniles during the whole year may indicate a longer reproductive period. The stomach content analysis identified T. marmorata as a top predator (TROPH=4.38) with a very narrow trophic niche (Bi=0.22), extremely specialized in feeding on bony fishes. In addition, this work provides new and updated information on T. torpedo and the rare Tetronarce nobiliana growth, reproductive features and diet. Therefore, offering new essential data to assess the population conditions of all Torpedinidae species that inhabit the Mediterranean Sea, will lead to effective conservation plans.

First Integrative Morphological and Genetic Characterization of Tremoctopus violaceussensu stricto in the Mediterranean Sea.

An integrative approach based on morphological and genetic analyses was undertaken for the first time to confirm the species identification of Mediterranean samples belonging to the genus Tremoctopus. Sequences of two mtDNA genes (cytochrome c oxidase subunit (COI) and 16S) were generated for the first time from Mediterranean samples. Both the similarity-based identifications and tree-based methods indicated that three females can be identified as Tremoctopus violaceus sensu stricto in agreement with their morphological classifications. All Mediterranean sequences clustered with the sequences of Tremoctopus violaceus from the Gulf of Mexico and were clearly differentiated from the sequences attributed to T. gracilis and T. robsoni. The chromatic pattern of the web and some features of gill filaments, arms formula, stylets, radulae, beaks, and stomach contents were given for all the samples; 105,758, 20,140, and 11,237 oocytes were estimated in the mature, immature, and developing samples, respectively. The presence of four spermatangia inside the cavity of the maturing female suggested the ability of this species to mate before reaching full maturity with more partners. Age investigation using beaks, performed for the first time in T. violaceus and within the genus gave results consistent with the different sizes and maturity conditions of the samples.

Living naked: first case of lack of skin-related structures in an elasmobranch, the blackmouth catshark (Galeus melastomus).

As far as is known, in this paper the first case of lacking of skin-related structures (epidermis, stratum laxum, dermal denticles and teeth) in a free-swimming elasmobranch, the blackmouth catshark, Galeus melastomus, is reported. The individual was caught by trawl in Sardinian waters (central-western Mediterranean) in July 2019 at a depth of 500 m. Although this kind of morphological abnormality is potentially fatal, the observations suggested that the specimen was in good health and well developed.

Shelf-life and labels: A cheap dating tool for seafloor macro litter? Insights from MEDITS surveys in Sardinian sea.

The global plastic economy demands to reduce the flow of plastic into oceans and promote remedial actions for already accumulated seafloor litter. In this perspective, baseline levels of contamination and tools for dating litter items in order to assess the efficacy of those actions, are thus needed. In this note we discuss the utility of introducing the acquisition of shelf-life and labels features from litter items into already established standardized protocols such as the one proposed by MEDiterranean International Trawl Survey (MEDITS). Our investigation was conducted on 612 high resolution images of litter items collected in each haul, which was retrieved during 6 years of surveys (2013-2018) around Sardinian sea (central western Mediterranean). While for the majority of items (89%) expiration date or labels were not present or legible, over 50% of dated items were likely dumped at sea from a period <5 years. In this perspective, this sort of data could represent a useful tool for monitoring the effectiveness of input reduction actions that implicitly rely on the gradual reduction of recently dumped items.

Dumping to the abyss: single-use marine litter invading bathyal plains of the Sardinian margin (Tyrrhenian Sea).

This study reports data on benthic litter abundance, composition and distribution obtained during deep-sea trawl surveys conducted along the Sardinian continental margin down to the bathyal plain, at depths comprised from 740 to 1740 m. None of the investigated sites was litter free. Density ranged from a minimum of 4 to a maximum of >1300 litter items per km2, with a mean value of 258 ±â€¯59 items. Plastic accounted for 56% of the total collected items, followed by glass (24%), metal (10%). Most items, irrespectively of the category, were single-use items. Fish abundance in all of the investigated catches was significantly higher than the number of litter items, the weight of which was similar to the reared fish biomass. Our results confirm that anthropogenic waste has reached the deep Mediterranean Sea, and that the most recent EU legislation banning single-use plastic tools represents a timely and necessary measure.

Natural history and molecular evolution of demersal Mediterranean sharks and skates inferred by comparative phylogeographic and demographic analyses.

BACKGROUND: The unique and complex paleoclimatic and paleogeographic events which affected the Mediterranean Sea since late Miocene deeply influenced the distribution and evolution of marine organisms and shaped their genetic structure. Following the Messinian salinity crisis and the sea-level fluctuations during the Pleistocene, several Mediterranean marine species developed deep genetic differentiation, and some underwent rapid radiation. Here, we consider two of the most prioritized groups for conservation in the light of their evolutionary history: sharks and rays (elasmobranchs). This paper deals with a comparative multispecies analysis of phylogeographic structure and historical demography in two pairs of sympatric, phylogenetically- and ecologically-related elasmobranchs, two scyliorhinid catsharks (Galeus melastomus, Scyliorhinus canicula) and two rajid skates (Raja clavata, Raja miraletus). Sampling and experimental analyses were designed to primarily test if the Sicilian Channel can be considered as effective eco-physiological barrier for Mediterranean demersal sympatric elasmobranchs. METHODS: The phylogeography and the historical demography of target species were inferred by analysing the nucleotide variation of three mitochondrial DNA markers (i.e., partial sequence of COI, NADH2 and CR) obtained from a total of 248 individuals sampled in the Western and Eastern Mediterranean Sea as well as in the adjacent northeastern Atlantic Ocean. Phylogeographic analysis was performed by haplotype networking and testing spatial genetic differentiation of samples (i.e., analysis of molecular variance and of principal components). Demographic history of Mediterranean populations was reconstructed using mismatch distribution and Bayesian Skyline Plot analyses. RESULTS: No spatial genetic differentiation was identified in either catshark species, while phylogeographic structure of lineages was identified in both skates, with R. miraletus more structured than R. clavata. However, such structuring of skate lineages was not consistent with the separation between Western and Eastern Mediterranean. Sudden demographic expansions occurred synchronously during the upper Pleistocene (40,000-60,000 years ago) in both skates and G. melastomus, likely related to optimal environmental conditions. In contrast, S. canicula experienced a slow and constant increase in population size over the last 350,000 years. DISCUSSION: The comparative analysis of phylogeographic and historical demographic patterns for the Mediterranean populations of these elasmobranchs reveals that historical phylogeographic breaks have not had a large impact on their microevolution. We hypothesize that interactions between environmental and ecological/physiological traits may have been the driving force in the microevolution of these demersal elasmobranch species in the Mediterranean rather than oceanographic barriers.

Amount and distribution of benthic marine litter along Sardinian fishing grounds (CW Mediterranean Sea).

Reports of marine litter pollution first appeared in scientific literature of the early 1970s; yet, more than 40 years later, no rigorous estimates exist of the amount of litter existing in the marine environment. To cope with this global urgency, this study reports the status of marine litter abundance along fishing grounds surrounding the island of Sardinia (CW Mediterranean Sea; FAO Geographical Sub-Area 11) through three years of trawl surveys. A total of 302 hauls, covering a total of 18.4 km2 of trawled surface were carried out in the framework of the MEDITS campaign, at depths comprised between 0 and 800 m. A total of 918 items were collected and sorted, with the highest concentration observed above 200 m depth. Overall, plastic was the dominant component of litter, followed by glass and metal. Comparing our results with other areas from the Mediterranean basin, Sardinian waters showed a lower impact, possibly as a consequence of multiple factors such as the lower human population density and the low flow of the main rivers, among others. In addition, fishermen behaviour with respect to marine litter was investigated by mean of anonymous questionnaires, emphasizing the necessity to further develop management policies and infrastructures supporting litter disposal.

Oviducal gland microstructure of Raja miraletus and Dipturus oxyrinchus (Elasmobranchii, Rajidae).

We studied the morphology and histology of the oviducal gland (OG) in the brown ray (Raja miraletus) and the long-nosed skate (Dipturus oxyrinchus) to understand its functional role in the reproductive strategy of these species. The external morphology of the gland was similar in both species, with lateral extensions like those found in other members of the Rajidae. Microscopic analysis showed a similar internal organization in both species. Immature and developing glands did not react to histochemical techniques. On reaching maturity, the OG had the largest width due to an increase in the production of secretory materials. In both species, the club zone of the gland showed a strong reaction to Periodic acid-Schiff (PAS) and alcian blue (AB) stains, indicating production of neutral and sulfated acid mucins. The secretory material produced by the papillary zone varied greatly between the two species. Both displayed tubular glands similar to those observed in the club zone, but in D. oxyrinchus the region near the lumen was intensely PAS+, whereas the last row of tubules of the brown ray stained intensely for a mixture of neutral and sulfated mucins. The baffle zone was the most conspicuous and extensive segment of all OGs, and it did not react to PAS/AB. The terminal zone, which is responsible for production of hair filaments, differed between the two species in terms of composition and organization of serous and mucous glands. This difference probably is related to the different substrates in which they release the egg capsules. Individual sperm detected in the brown ray baffle lamellae could be the result of a recent mating, whereas their presence in the deep recesses of the baffle and in the terminal zone of the long-nosed skate might indicate sperm storage.