Gluconeogenesis in the yolk syncytial layer-like tissue of cloudy catshark (Scyliorhinus torazame).

Glucose has important roles in the development of zebrafish, the vertebrate animal model; however, in most oviparous animals, the amount of maternally provided glucose in the yolk is scarce. For these reasons, developing animals need some ways to supplement glucose. Recently, it was found that developing zebrafish, a teleost fish, undergo gluconeogenesis in the yolk syncytial layer (YSL), an extraembryonic tissue that surrounds the yolk. However, teleost YSL is evolutionarily unique, and it is not clear how other vertebrates supplement glucose. In this study, we used cloudy catshark (or Torazame catshark), an elasmobranch species which possesses a YSL-like tissue during development, and sought for possible gluconeogenic activities in this tissue. In their yolk sac, glucose increased, and our isotope tracking analysis detected gluconeogenic activities with glycerol most preferred substrate. In addition, many of gluconeogenic genes were expressed at the YSL-like tissue, suggesting that cloudy catshark engages in gluconeogenesis in this tissue. The gluconeogenesis in teleost YSL and a similar tissue in elasmobranch species implies conserved mechanisms of yolk metabolism between these two lineages. Future studies on other vertebrate taxa will be helpful to understand the evolutionary changes in the modes of yolk metabolism that vertebrates have experienced.

Interference with the retinoic acid signalling pathway inhibits the initiation of teeth and caudal primary scales in the small-spotted catshark Scyliorhinus canicula.

The retinoic acid (RA) pathway was shown to be important for tooth development in mammals, and suspected to play a key role in tooth evolution in teleosts. The general modalities of development of tooth and "tooth-like" structures (collectively named odontodes) seem to be conserved among all jawed vertebrates, both with regard to histogenesis and genetic regulation. We investigated the putative function of RA signalling in tooth and scale initiation in a cartilaginous fish, the small-spotted catshark Scyliorhinus canicula. To address this issue, we identified the expression pattern of genes from the RA pathway during both tooth and scale development and performed functional experiments by exposing small-spotted catshark embryos to exogenous RA or an inhibitor of RA synthesis. Our results showed that inhibiting RA synthesis affects tooth but not caudal primary scale development while exposure to exogenous RA inhibited both. We also showed that the reduced number of teeth observed with RA exposure is probably due to a specific inhibition of tooth bud initiation while the observed effects of the RA synthesis inhibitor is related to a general delay in embryonic development that interacts with tooth development. This study provides data complementary to previous studies of bony vertebrates and support an involvement of the RA signalling pathway toolkit in odontode initiation in all jawed vertebrates. However, the modalities of RA signalling may vary depending on the target location along the body, and depending on the species lineage.

Distribution of Brain-Derived Neurotrophic Factor in the Brain of the Small-Spotted Catshark Scyliorhinus canicula, and Evolution of Neurotrophins in Basal Vertebrates.

Neurotrophins (NTFs) are structurally related neurotrophic factors essential for differentiation, survival, neurite outgrowth, and the plasticity of neurons. Abnormalities associated with neurotrophin-signaling (NTF-signaling) were associated with neuropathies, neurodegenerative disorders, and age-associated cognitive decline. Among the neurotrophins, brain-derived neurotrophic factor (BDNF) has the highest expression and is expressed in mammals by specific cells throughout the brain, with particularly high expression in the hippocampus and cerebral cortex. Whole genome sequencing efforts showed that NTF signaling evolved before the evolution of Vertebrates; thus, the shared ancestor of Protostomes, Cyclostomes, and Deuterostomes must have possessed a single ortholog of neurotrophins. After the first round of whole genome duplication that occurred in the last common ancestor of Vertebrates, the presence of two neurotrophins in Agnatha was hypothesized, while the monophyletic group of cartilaginous fishes, or Chondrichthyans, was situated immediately after the second whole genome duplication round that occurred in the last common ancestor of Gnathostomes. Chondrichthyans represent the outgroup of all other living jawed vertebrates (Gnathostomes) and the sister group of Osteichthyans (comprehensive of Actinopterygians and Sarcopterygians). We were able to first identify the second neurotrophin in Agnatha. Secondly, we expanded our analysis to include the Chondrichthyans, with their strategic phylogenetic position as the most basal extant Gnathostome taxon. Results from the phylogenetic analysis confirmed the presence of four neurotrophins in the Chondrichthyans, namely the orthologs of the four mammalian neurotrophins BDNF, NGF, NT-3, and NT-4. We then proceeded to study the expression of BDNF in the adult brain of the Chondrichthyan Scyliorhinus canicula. Our results showed that BDNF is highly expressed in the S. canicula brain and that its expression is highest in the Telencephalon, while the Mesencephalic and Diencephalic areas showed expression of BDNF in isolated and well-defined cell groups. NGF was expressed at much lower levels that could be detected by PCR but not by in situ hybridization. Our results warrant further investigations in Chondrichthyans to characterize the putative ancestral function of neurotrophins in Vertebrates.

Progesterone initiates tendril formation in the oviducal gland during egg encapsulation in cloudy catshark (Scyliorhinus torazame).

The diverse reproductive strategies of elasmobranchs (sharks, rays, and skates) have attracted research attention, but the endocrine control of reproduction is still incompletely known in elasmobranchs. By long-term monitoring of the egg-laying cycle in cloudy catsharks (Scyliorhinus torazame), we recently demonstrated a transient increase in plasma progesterone (P4) levels just prior to the appearance of the capsulated eggs in the oviducts. In the present study, we examined the in vivo effects of P4 administration in mature female cloudy catsharks. Although no capsulated eggs were observed following the implantation of P4-containing silicone tubing, we did find dark swollen oviducts in the abdominal cavity, in which clumps of long and coiled tendrils were observed. The tendril is an extension of the egg capsule, and the formation of the egg capsule begins with the tendril before main capsule formation. During the period of P4 implantation, the tendrils elongated, and their diameters were significantly increased on day 2 of treatment. Tendril formation was also confirmed on the day of endogenous P4 surge. Tendrils were not formed in catsharks implanted with estradiol-17ß or testosterone. Histological analysis of the oviducal gland revealed that P4 implantation induced the secretion of tendril materials from the secretory tubules in the baffle zone, while the tendril materials were stored in the cytoplasm of the secretory cells under low P4 condition. Morphometrically, the area of secreted luminal materials in the secretory tubules was highly correlated to the diameter of tendrils. Our results suggest that the P4 surge during the egg-laying cycle serves as a trigger for egg capsule formation in the oviducal gland of cloudy catshark, but the hormonal signals were incomplete as the main capsule was not formed. Further studies are required to identify the hormones required for ovulation and formation of the main egg capsule.

Localization and Characterization of Major Neurogenic Niches in the Brain of the Lesser-Spotted Dogfish Scyliorhinus canicula.

Adult neurogenesis is defined as the ability of specialized cells in the postnatal brain to produce new functional neurons and to integrate them into the already-established neuronal network. This phenomenon is common in all vertebrates and has been found to be extremely relevant for numerous processes, such as long-term memory, learning, and anxiety responses, and it has been also found to be involved in neurodegenerative and psychiatric disorders. Adult neurogenesis has been studied extensively in many vertebrate models, from fish to human, and observed also in the more basal cartilaginous fish, such as the lesser-spotted dogfish, Scyliorhinus canicula, but a detailed description of neurogenic niches in this animal is, to date, limited to the telencephalic areas. With this article, we aim to extend the characterization of the neurogenic niches of S. canicula in other main areas of the brain: we analyzed via double immunofluorescence sections of telencephalon, optic tectum, and cerebellum with markers of proliferation (PCNA) and mitosis (pH3) in conjunction with glial cell (S100ß) and stem cell (Msi1) markers, to identify the actively proliferating cells inside the neurogenic niches. We also labeled adult postmitotic neurons (NeuN) to exclude double labeling with actively proliferating cells (PCNA). Lastly, we observed the presence of the autofluorescent aging marker, lipofuscin, contained inside lysosomes in neurogenic areas.

A new parasitic copepod species of Tautochondria (Siphonostomatoida: Hyponeoidae), found in the narrowmouthed catshark Schroederichthys bivius from the Strait of Magellan.

Hyponeoidae is a copepod family that is rare around the world, with only three species described until now. Recently, a hyponeoid copepod was found in the narrowmouthed catshark, Schroederichthys bivius, from Chile, which has not been formally described. The objective of this study is to describe morphologically a new species of Hyponeoidae, from samples of the Strait of Magellan, Southern Chile. Also, a genetic analysis, based on the COI gene, was used to determine the relatedness of the new hyponeoid species to other copepods from Siphonostomatoida, which were available in the GenBank platform. The new species belongs to the genus Tautochondria and is here described as T. magellanica n. sp. This species differs from T. dolichoura mainly in the presence of long process at each side of the buccal cone, the absence of lobes on the head and short processes on the genital complex. According to the genetic sequences, T. magellanica n. sp. was not closely related to any other species. Therefore, this result confirms that Hyponeoidae is a separate family. However, the relatedness to other genera in Siphonostomatoida is still unknown.

Increasing the data on elasmobranch plasma protein electrophoresis: electrophoretogram reference values determination in the undulate skate (Raja Undulata) and the nursehound shark (Scyliorhinus stellaris) maintained under human care.

BACKGROUND: This study determined plasma protein electrophoresis (PPE) reference intervals in two elasmobranch species: the undulate skate (Raja undulata) and the nursehound shark (Scyliorhinus stellaris), using a reference population of 48 undulate skates (27 males, 21 females) and 62 nursehounds (32 males, 30 females), considered to be clinically healthy. Plasma samples were analyzed using capillary zone electrophoresis (CZE). RESULTS: The undulate skate electrophoretogram resembled those previously reported in other batoids and could be divided into seven consistent fractions. No statistically significant differences were detected between sexes and developmental stages. The nursehound electrophoretogram was similar to that previously described in other shark species and could be divided into eight consistent fractions. Fraction 5% was significantly higher in juvenile nursehounds when compared to adults, while fraction 6 concentration and percentage were significantly higher in adults. Fraction 4% was higher in males than in females. Albumin band was not detected, and pre-albumin was negligible in both studied species. Alpha-globulins were predominant in the undulate skate, while beta-globulins were predominant in nursehounds. Statistically significant differences were found in all electrophoretogram fraction percentages and concentrations between the two species. CONCLUSION: To the authors knowledge, this is the first study reporting PPE values in undulate skates and nursehounds, and the first study using CZE in elasmobranch plasma. These findings can serve as a primary reference for health monitoring in both species and will add to the limited data available on PPE in elasmobranchs.

Developmental genoarchitectonics as a key tool to interpret the mature anatomy of the chondrichthyan hypothalamus according to the prosomeric model.

The hypothalamus is a key vertebrate brain region involved in survival and physiological functions. Understanding hypothalamic organization and evolution is important to deciphering many aspects of vertebrate biology. Recent comparative studies based on gene expression patterns have proposed the existence of hypothalamic histogenetic domains (paraventricular, TPa/PPa; subparaventricular, TSPa/PSPa; tuberal, Tu/RTu; perimamillary, PM/PRM; and mamillary, MM/RM), revealing conserved evolutionary trends. To shed light on the functional relevance of these histogenetic domains, this work aims to interpret the location of developed cell groups according to the prosomeric model in the hypothalamus of the catshark Scyliorhinus canicula, a representative of Chondrichthyans (the sister group of Osteichthyes, at the base of the gnathostome lineage). To this end, we review in detail the expression patterns of ScOtp, ScDlx2, and ScPitx2, as well as Pax6-immunoreactivity in embryos at stage 32, when the morphology of the adult catshark hypothalamus is already organized. We also propose homologies with mammals when possible. This study provides a comprehensive tool to better understand previous and novel data on hypothalamic development and evolution.

Investigating the effects of microplastic ingestion in Scyliorhinus canicula from the South of Sicily.

Plastic pollution is increasing dramatically worldwide, causing adverse effects on a wide variety of marine organisms at all trophic levels. As predators, sharks play a key role in marine ecosystems and they could especially be threatened by the ingestion of microplastics. This study contributes to expand the existing data on the MPs ingestion by a Mediterranean elasmobranch species, Scyliorhinus canicula, adding new information on the potential impact that this class of contaminants can have on the metabolism of this ecologically relevant species. The present research is aimed: i) to assess if the ingestion of MPs in S. canicula is related to sex, size and depth of catch; ii) to evaluate the effect of MPs on fish body condition; iii) to evaluate if the ingested MPs influence the amino acid and fatty acid composition of eye and liver. A total of 61 specimens of S. canicula were analysed. Forty-nine individuals (80.3 %) had ingested plastic items. Totally, 147 plastic elements were found, mainly belonging to small MPs (49 %), and large MPs (46 %), mostly represented by fibers (84 %). The predominant colour was black. No differences were found between sex and size. A difference in the number of items/specimens related to the deep, highest between 50 and 100 m (4.4), while the lowest between 101 and 500 m (2.1) was found. The condition factor (Kn) value was equal to 1.00 highlighting the wellness of the fish. Arginine (20.1 %), Glutamate (17.4), Phenylalanine (15.7 %), Proline (15.6 %) and were the most abundant amino acids in the eyes of S. canicula. The relative fatty acid composition of the livers was dominated by fatty acids SA (30.2 %) (SFA), CA (29.9 %) (SFA) and OA (22.4 %) (MUFA). This paper reports a study on the relationship between amino acids and fatty acids composition and ingested MPs, highlighting that no significant effects were found.

Long-term monitoring of egg-laying cycle using ultrasonography reveals the reproductive dynamics of circulating sex steroids in an oviparous catshark, Scyliorhinus torazame.

The many diverse reproductive strategies of elasmobranchs (sharks, skates and rays) from lecithotrophic oviparity to matrotrophic viviparity have attracted significant research attention. However, the endocrine control of elasmobranch reproduction is less well-documented largely due to their reproductive characteristics, such as a long reproductive cycle, and/or repeated internal fertilization using stored sperm in oviparous species. In the present study, for the first time, we succeeded in non-invasive monitoring of the continuing egg-laying cycle of the cloudy catshark Scyliorhinus torazame using portable ultrasound devices. Furthermore, long-term simultaneous monitoring of the egg-laying cycle and measurement of plasma sex steroids revealed cycling patterns of estradiol-17ß (E2), testosterone (T) and progesterone (P4). In particular, a decline in T followed by a reciprocal surge in plasma P4 were consistently observed prior to the appearance of the capsulated eggs, implying that P4 is likely associated with the ovulation and/or egg-case formation. While the cycling pattern of E2 was not as apparent as those of T and P4, threshold levels of E2 (>5 ng/mL) and T (>1 ng/mL) appeared to be crucial in the continuation of egg-laying cycle. The possibility to trace the dynamics of plasma sex steroids in a single individual throughout the reproductive cycles makes the catshark a useful model for regulatory and mechanistic studies of elasmobranch reproduction.

Evolutionary Transition in the Regulation of Vertebrate Pronephros Development: A New Role for Retinoic Acid.

The anterior-posterior (AP) axis in chordates is regulated by a conserved set of genes and signaling pathways, including Hox genes and retinoic acid (RA), which play well-characterized roles in the organization of the chordate body plan. The intermediate mesoderm (IM), which gives rise to all vertebrate kidneys, is an example of a tissue that differentiates sequentially along this axis. Yet, the conservation of the spatiotemporal regulation of the IM across vertebrates remains poorly understood. In this study, we used a comparative developmental approach focusing on non-conventional model organisms, a chondrichthyan (catshark), a cyclostome (lamprey), and a cephalochordate (amphioxus), to assess the involvement of RA in the regulation of chordate and vertebrate pronephros formation. We report that the anterior expression boundary of early pronephric markers (Pax2 and Lim1), positioned at the level of somite 6 in amniotes, is conserved in the catshark and the lamprey. Furthermore, RA, driving the expression of Hox4 genes like in amniotes, regulates the anterior pronephros boundary in the catshark. We find no evidence for the involvement of this regulatory hierarchy in the AP positioning of the lamprey pronephros and the amphioxus pronephros homolog, Hatschek's nephridium. This suggests that despite the conservation of Pax2 and Lim1 expressions in chordate pronephros homologs, the responsiveness of the IM, and hence of pronephric genes, to RA- and Hox-dependent regulation is a gnathostome novelty.

Early shape divergence of developmental trajectories in the jaw of galeomorph sharks.

BACKGROUND: The onset of morphological differences between related groups can be tracked at early stages during embryological development. This is expressed in functional traits that start with minor variations, but eventually diverge to defined specific morphologies. Several processes during this period, like proliferation, remodelling, and apoptosis for instance, can account for the variability observed between related groups. Morphological divergence through development is often associated with the hourglass model, in which early stages display higher variability and reach a conserved point with reduced variability from which divergence occurs again to the final phenotype. RESULTS: Here we explored the patterns of developmental shape changes in the lower jaw of two shark species, the bamboo shark (Chiloscyllium punctatum) and the catshark (Scyliorhinus canicula). These two species present marked differences in their foraging behaviour, which is reflected in their adult jaw morphology. By tracing the developmental sequence of the cartilage condensation, we identified the onset of cartilage for both species at around stage 31. Other structures that developed later without a noticeable anlage were the labial cartilages, which appear at around stage 33. We observed that the lower jaw displays striking differences in shape from the earliest moments, without any overlap in shape through the compared stages. CONCLUSIONS: The differences observed are also reflected in the functional variation in feeding mechanism between both species. Likewise, the trajectory analysis shows that the main differences are in the magnitude of the shape change through time. Both species follow a unique trajectory, which is explained by the timing between stages.

When Bigger Is Better: 3D RNA Profiling of the Developing Head in the Catshark Scyliorhinus canicula.

We report the adaptation of RNA tomography, a technique allowing spatially resolved, genome-wide expression profiling, to a species occupying a key phylogenetic position in gnathostomes, the catshark Scyliorhinus canicula. We focused analysis on head explants at an embryonic stage, shortly following neural tube closure and of interest for a number of developmental processes, including early brain patterning, placode specification or the establishment of epithalamic asymmetry. As described in the zebrafish, we have sequenced RNAs extracted from serial sections along transverse, horizontal and sagittal planes, mapped the data onto a gene reference taking advantage of the high continuity genome recently released in the catshark, and projected read counts onto a digital model of the head obtained by confocal microscopy. This results in the generation of a genome-wide 3D atlas, containing expression data for most protein-coding genes in a digital model of the embryonic head. The digital profiles obtained for candidate forebrain regional markers along antero-posterior, dorso-ventral and left-right axes reproduce those obtained by in situ hybridization (ISH), with expected relative organizations. We also use spatial autocorrelation and correlation as measures to analyze these data and show that they provide adequate statistical tools to extract novel expression information from the model. These data and tools allow exhaustive searches of genes exhibiting any predefined expression characteristic, such a restriction to a territory of interest, thus providing a reference for comparative analyses across gnathostomes. This methodology appears best suited to species endowed with large embryo or organ sizes and opens novel perspectives to a wide range of evo-devo model organisms, traditionally counter-selected on size criterion.

The complete mitogenome of the blackgill catshark, Parmaturus melanobranchus (Chan, 1966).

A rare specimen of Parmaturus melanobranchus was collected from the South China Sea. The complete mitochondrial genome of the specimen was sequenced using Illumina Hiseq platform and assembled with Geneious and Trinity. The mitogenome is 16,687 bp long with a base composition of 30.4% A, 14.1% G, 23.5% C and 32.1% T, respectively. A total of 37 genes were predicted containing 13 protein-coding genes, 2 ribosomal RNA genes and 22 transfer RNA genes. This is the first complete mitochondrial genome published of this species.

Evolution of lbx spinal cord expression and function.

Ladybird homeobox (Lbx) transcription factors have crucial functions in muscle and nervous system development in many animals. Amniotes have two Lbx genes, but only Lbx1 is expressed in spinal cord. In contrast, teleosts have three lbx genes and we show here that zebrafish lbx1a, lbx1b, and lbx2 are expressed by distinct spinal cell types, and that lbx1a is expressed in dI4, dI5, and dI6 interneurons, as in amniotes. Our data examining lbx expression in Scyliorhinus canicula and Xenopus tropicalis suggest that the spinal interneuron expression of zebrafish lbx1a is ancestral, whereas lbx1b has acquired a new expression pattern in spinal cord progenitor cells. lbx2 spinal expression was probably acquired in the ray-finned lineage, as this gene is not expressed in the spinal cords of either amniotes or S. canicula. We also show that the spinal function of zebrafish lbx1a is conserved with mouse Lbx1. In zebrafish lbx1a mutants, there is a reduction in the number of inhibitory spinal interneurons and an increase in the number of excitatory spinal interneurons, similar to mouse Lbx1 mutants. Interestingly, the number of inhibitory spinal interneurons is also reduced in lbx1b mutants, although in this case the number of excitatory interneurons is not increased. lbx1a;lbx1b double mutants have a similar spinal interneuron phenotype to lbx1a single mutants. Taken together these data suggest that lbx1b and lbx1a may be required in succession for correct specification of dI4 and dI6 spinal interneurons, although only lbx1a is required for suppression of excitatory fates in these cells.

New monogeneans from the bathydemersal southern African endemic catshark, Holohalaelurus regani (Gilchrist, 1922).

The catshark genus Holohalaelurus Fowler is currently represented by five species distributed off the southern and eastern parts of the African coast. Very few parasitological records exist for any of these five species, representing a significant knowledge gap. We report the first monogenean species from the bathydemersal species Holohalaelurus regani (Gilchrist): a new species of Microbothriidae Price, 1936, Leptomicrobothrium holohalaelure n. sp. from the dorsal skin surface, and a new species of Hexabothriidae Price, 1942 representing a new genus, Scyliorhinocotyle narvaezae n. gen., n. sp., from the gill lamellae. Both monogenean records represent the first for any member of the catshark genus Holohalaelurus. Previously we focused on the identification of monogenean taxa of emerging veterinary importance for public aquaria. We now begin a focused effort to document a generally unexplored monogenean biodiversity from diverse marine habitats off South Africa.

Plastic Ingestion by the Small-Spotted Catshark (Scyliorhinus canicula) from the South West Coast of the United Kingdom.

Plastic pollution has increased dramatically worldwide and is having a detrimental effect on a variety of marine organisms. This study contributes to the existing data on the ingestion of both macroplastics and microplastics by one elasmobranch species, the small-spotted catshark (Scyliorhinus canicula), considering the occurrence in males and females. Plastics were extracted from the stomach and spiral valve of (N = 200, n = 100 females and n = 100 males) and analysed by visual observations. A total of 28 pieces of plastic were found with five pieces extracted from a single individual. Overall findings indicate that this shark species is vulnerable to some extent to plastic ingestion (6.5% frequency of occurrence), but different aspects, such as differences in plastic contamination levels of different locations and ability to regurgitate stomach content, may affect the results of similar studies.

Biological Sound vs. Anthropogenic Noise: Assessment of Behavioural Changes in Scyliorhinus canicula Exposed to Boats Noise.

Despite the growing interest in human-made noise effects on marine wildlife, few studies have investigated the potential role of underwater noise on elasmobranch species. In this study, twelve specimens of small-spotted catshark (Scyliorhinus canicula) were exposed to biological and anthropogenic sounds in order to assess their behavioural changes in response to prey acoustic stimuli and to different amplitude levels of shipping noise. The sharks, individually held in aquariums, were exposed to four experimental acoustic conditions characterized by different spectral (Hz) components and amplitude (dB re 1 µPa) levels. The swimming behaviour and spatial distribution of sharks were observed. The results highlighted significant differences in swimming time and in the spatial use of the aquarium among the experimental conditions. When the amplitude levels of biological sources were higher than those of anthropogenic sources, the sharks' swimming behaviour was concentrated in the bottom sections of the aquarium; when the amplitude levels of anthropogenic sources were higher than biological ones, the specimens increased the time spent swimming. Moreover, their spatial distribution highlighted a tendency to occupy the least noisy sections of the aquarium. In conclusion, this study highlighted that anthropogenic noise is able to affect behaviour of catshark specimens and the impact depends on acoustic amplitude levels.

Adverse effects of plastic ingestion on the Mediterranean small-spotted catshark (Scyliorhinus canicula).

Plastics are widely diffused in the oceans and their ingestion by marine organisms is raising concern for potentially adverse effects. The risk of harmful interactions with marine plastic pollution depends on the biology of the species as well as the distribution and abundance of the different plastic types. The aim of this study was to assess the occurrence of plastic ingestion by the small-spotted catshark (Scyliorhinus canicula), one of the most abundant elasmobranchs in the Mediterranean Sea. The expression levels of genes indicative of total immune system function were analyzed to gather preliminary data for further investigation of any potential correlations between plastic presence and immune activation. One hundred catsharks were collected during the Spring 2018 in two geographic locations in the southern region of the central Mediterranean Sea: 1) near Mazara del Vallo, SW Sicily and 2) near Lampedusa island, Italy's southernmost. Standard measurements were recorded for each specimen and its organs and sex was determined. The gastrointestinal tract (GIT) was preserved for plastic detection and identification. Where present, plastics (macro- and micro-) were characterized in terms of size, shape and polymer typology through microscopy and µ-Raman spectroscopy. Spleen from a subset of thirty samples was preserved for RNA extraction, then used to quantify by real time PCR the transcripts of T cell receptor beta (TCRB), T cell receptor delta (TCRD) and IgM genes. The results indicated that ingestion of plastic is widespread, with microplastics (MP, from 1 µm to <1 mm) abundantly present in nearly all samples and macroplasticplastic (MaP, > 1 cm) in approximately 18% of the specimens collected. A significant increase in the expression of TCRB, TCRD and IgM was observed in the spleen of MaP + specimens from Mazara del Vallo waters, in parallel with 67% increase in liver weight. While the presence of MP alone is not enough to induce a strong activation of the immunity, some type of plastics falling into the MaP category may be more toxic than others and crucial in the activation of the immune response. The results of this study represent a first evidence that plastic pollution represents an emerging threat to S. canicula, the Mediterranean food web and human consumers.

A new genus and species of catshark (Carcharhiniformes: Scyliorhinidae) from eastern Indonesia.

A new genus and species of catshark is described based on a single specimen collected off Ambon in the Maluku Islands of eastern Indonesia. Akheilos suwartanai belongs to the subfamily Schroederichthyinae which differs from the other catsharks in a combination of: similar sized dorsal fins, supraorbital crests present, pseudosiphon present on claspers, broad subocular ridges under eyes, posterior nasal flaps present, tips of rostral cartilage fused into a rostral node. It represents the first record of this subfamily outside of the Americas. Akheilos differs from the other genus in the subfamily, Schroederichthys in a combination of: clasper groove not fused dorsally, ventral lobe of caudal fin produced, more intestinal valve turns, anal fin slightly larger than second dorsal fin, and in colour pattern.