Identification of possible stock units of Anguilla anguilla through microchemical and morphometric otolith analyses

L’anguilla europea, è una specie eurialina catadroma con un complesso ciclo biologico: l’area di riproduzione, unica, si trova molto distante da quella di distribuzione. La specie necessita di una gestione dello stock a fini conservazionistici. Il problema è europeo: lo stock è unico, distribuito in Europa e nell’Africa settentrionale, si riproduce in Atlantico ed è panmittico. C’è preoccupazione per il declino del reclutamento e delle catture di adulti. Lo scopo del progetto è di individuare possibili unità di stock nella penisola italiana. La ricerca è basata sullo studio degli otoliti mediante analisi morfometrica e microchimica. I contorni degli otoliti sono sottoposti ad analisi ellittica di Fourier per individuare eventuali gruppi. Gli otoliti sono stati levigati per effettuare: letture d’età, indagini microstrutturali al SEM delle fasi larvali, analisi microchimiche LA-ICP-MS del nucleo, studiarne l’origine e valutare l’ambiente di sviluppo. Le indagini morfometriche mostrano evidenti pattern ontogenetici, ma non legati ocorrelati alla località, sesso o anno di nascita. Le indagini microstrutturali hanno evidenziano l’alto contenuto organico nucleare, un pattern comune di crescita ed eventi chiave delle fasi larvali, con una media di 212 anelli giornalieri. La microchimica rivela che le larve si sviluppano in acque salate fino alla metamorfosi, poi migrano verso acque meno salate. Le analisi su campioni nati nello stesso anno, evidenziano due gruppi: individui di rimonta naturale e individui di ripopolamento. I profili nucleo bordo evidenziano la permanenza a salinità intermedie degli adulti. L’attività di ricerca si è dimostrata proficua dal punto di vista tecnico con la messa a punto di protocolli innovativi e con forti ricadute sulla riduzione dei tempi e costi d’analisi. Il debole segnale di possibili unità di stock andrà verificato in futuro mediante analisi più dettagliate discriminando meglio la storia di ogni singolo individuo.The European eel is euryhaline catadromous with a complex life cycle: spawning area, unique, is far from that of distribution. The species needs a stock management and its conservation. The problem is European: the stock is unique, distributed in Europe and North Africa, spawning in Atlantic Sea and is panmictic. There is concern about the decline of recruitment and catches of adults. The purpose of the project is to identify possible units of stock in the Italian peninsula. The research is based on the study of otoliths by morphometric and microchemical analyses . The otoliths contours are analyzed by elliptic Fourier function to discover any groups. Otoliths were polished to make: ageing, SEM microstructural investigations of larval stages, LA-ICP-MS microchemical analysis of the core to study the origin and along a transect core to edge to evaluate the development environment. Morphometric investigations show obvious ontogenetic pattern, but no location, gender or year of birth correlation. The microstructural investigations showed high organic content in the core, a common pattern of growth and key events of the larval stages, with an average of 212 daily rings. The microchemical reveals that the larvae develop into salt water until metamorphosis, then migrate to waters less salty. Analyses of samples born in the same year, showed two groups: wild and restocked. The profiles confirm that the adult stay in brackishwater. The development of innovative protocols and improved technics have guaranteed a strong reduction of time and cost of analysis. The weak signal of possible stock units will be further tested through more detailed analysis discriminating about the life history of each individual

Distribution and some biological parameters of the red gurnard, Chelidonichthys cuculus (Actinopterygii, Scorpaeniformes, Triglidae) in the north-central Adriatic Sea

Background. Red gurnard, Chelidonichthys cuculus (Linnaeus, 1758), is distributed in the Mediterranean Sea, Black Sea, and in the eastern Atlantic Ocean from Norway to Mauritania, although it is rarely found in the North Sea and on the Norwegian shelf. The aim of this work was to conduct a comprehensive study of red gurnard in the Adriatic Sea on selected aspects the population biology and some of its environmental implications . Materialsandmethods. The sampling covered the entire subdivision GSA17 (north-central Adriatic Sea) and it was based on several bottom trawl surveys of MEDITS and GRUND projects from 2000 through 2009. Biometric parameters, as well as the spatial- and depth distributions were analyzed. The parameters such as the length–weight relation, sex, gonad maturity stage, and the stomach content were determined on a subsample of 539 specimens collected seasonally (summer and autumn–winter) within 2007–2009. The data were processed statistically . Results. Chelidonichthys cuculus was mainly distributed in the central Adriatic. Juveniles showed a distribution in shallower waters while adults tended to migrate towards the Croatian coast at greater depths. The total length of the fish sampled ranged from 40 to 303 mm (TL). We assumed the length of 100 mm as a threshold separat ing juveniles from adults and the length range of 170–190 mm as the maturity size for females and males. Isometric- and allometric growth patterns were showed for the juveniles and the adults, respectively. The diet was based almost exclusively on crustaceans (IRI% > 98%) and it did not change upon reaching the sexual maturity. Chelidonichthys cuculus should be considered a specialist predator feeding mostly on necto-benthic lophogastrids (IRI% > 72%) . Conclusion. This work constitutes an attempt to summarize some environmental and biology aspects of the red gurnard in Adriatic population: differences in depth distribution and growth were found between juveniles and adults but not in diet. Males and females showed differences in growth parameters. Extending the studied area and collecting data of red gurnard in the main areas of production should help in better understanding the biolo gy and dynamics of this species

Multiscale analysis on otolith structural features reveals differences in ontogenesis and sex in Merluccius merluccius in the western Adriatic Sea

Otolith biomineralization results from biochemical processes regulated by the interaction of internal (physiological) and external (environmental) factors which lead to morphological and ultrastructural variability at intra- and interspecific levels. The aim of this study was to conduct a multi-scale analysis of the sagittal otoliths of the Merlucius merlucius (European hake) from the western Adriatic Sea in order to correlate otolith features with fish ontogeny and sex. We show that otoliths of sexually undifferentiated (non-sexed) individuals having a fish body total length (TL) less than 15 cm had faster growth in length, width, area, perimeter, volume and weight and a higher amount of organic matrix compared with otoliths of sexually differentiated individuals (females and males) having a fish size range of 15-50 cm. Most importantly, with increasing fish TL, female saccular otoliths contained a higher number of protuberances and rougher surface compared with male specimens, which showed more uniform mean curvature density. The differences between females and males discovered in this study could be associated with fish hearing adaptation to reproductive behavioural strategies during the spawning season. The outcomes of this research provide insights on how size and sex-related variations in otolith features may be affected by fish ecological and behavioural patterns

To Be, or Not to Be: That Is the Hamletic Question of Cryptic Evolution in the Eastern Atlantic and Mediterranean Raja miraletus Species Complex

The ability to correctly identify specimens at the species level is crucial for assessing and conserving biodiversity. Despite this, species-specific data are lacking for many of South Africa’s catsharks due to a high level of morphological stasis. As comprehensive and curated DNA reference libraries are required for the reliable identification of specimens from morphologically similar species, this study reviewed and contributed to the availability of cytochrome c oxidase subunit I (COI) and nicotinamide adenine dehydrogenase subunit 2 (NADH2) sequences for South Africa’s catsharks. A molecular taxonomic approach, implementing species delimitation and specimen assignment methods, was used to assess and highlight any taxonomic uncertainties and/or errors in public databases. The investigated species were summarised into 47 molecular operational taxonomic units (MOTUs), with some conflicting specimen assignments. Two Apristurus specimens sampled in this study remained unidentified, revealing the presence of previously undocumented genetic diversity. In contrast, haplotype sharing within Haploblepharus—attributed to nucleotide ambiguities—resulted in the delimitation of three congeners into a single MOTU. This study reveals that molecular taxonomy has the potential to flag undocumented species and/or misidentified specimens, and further highlights the need to implement integrated taxonomic assessments on catsharks that represent an irreplaceable component of biodiversity in the region

Strongly structured populations and reproductive habitat fragmentation increase the vulnerability of the Mediterranean starry ray Raja asterias (Elasmobranchii, Rajidae)

1. The Mediterranean starry ray (Raja asterias) populations within the Mediterranean Sea are susceptible to high rates of bycatch in the multispecies trawl fisheries. Understanding its population structure and identifying critical habitats are crucial for assessing species vulnerability and setting the groundwork for specific management measures to prevent population decline. 2. To assess the population structure of R. asterias in the Mediterranean, the genetic variation in nine population samples at one mitochondrial marker and eight nuclear microsatellite loci was analysed. Moreover, 172 egg cases collected in the Strait of Sicily were identified at species level using integrated molecular and morphological approaches. 3. Genetic analyses revealed that the Mediterranean starry ray comprises three distinct units inhabiting the western, the central-western, and the central-eastern areas of the Mediterranean. An admixture zone occurs in the Strait of Sicily and the Ionian Sea, where individuals of the central-western and central-eastern population units intermingle. 4. The joint morphometric–genetic analyses of rajid egg cases confirmed the presence of more than one species in the admixture area, with a predominance of egg cases laid by R. asterias. DNA barcoding revealed that egg cases and embryos of R. asterias shared several haplotypes with adult individuals from the centralwestern and central-eastern Mediterranean Sea, revealing that females of both populations laid numerous eggs in this area. 5. According to these findings, detailed taxonomic determination of egg cases, when combined with seasonal migration studies, could improve the capability to identify important spawning or nursery areas for the Mediterranean starry ray, particularly in those admixture zones relevant to maintaining genetic diversity. 6. Finally, these new insights should be considered to update the Action Plan for the Conservation of Cartilaginous Fishes in the Mediterranean Sea with effective measures to reduce the impact of skate bycatch in trawling and safeguard egg cases in nursery areas

Genetic differentiation and phylogeography of Mediterranean-North Eastern Atlantic blue shark (Prionace glauca, L. 1758) using mitochondrial DNA: Panmixia or complex stock structure?

Background The blue shark (Prionace glauca, Linnaeus 1758) is one of the most abundant epipelagic shark inhabiting all the oceans except the poles, including the Mediterranean Sea, but its genetic structure has not been confirmed at basin and interoceanic distances. Past tagging programs in the Atlantic Ocean failed to find evidence of migration of blue sharks between the Mediterranean and the adjacent Atlantic, despite the extreme vagility of the species. Although the high rate of by-catch in the Mediterranean basin, to date no genetic study on Mediterranean blue shark was carried out, which constitutes a significant knowledge gap, considering that this population is classified as “Critically Endangered”, unlike its open-ocean counterpart. Methods Blue shark phylogeography and demography in the Mediterranean Sea and North-Eastern Atlantic Ocean were inferred using two mitochondrial genes (Cytb and control region) amplified from 207 and 170 individuals respectively, collected from six localities across the Mediterranean and two from the North-Eastern Atlantic. Results Although no obvious pattern of geographical differentiation was apparent from the haplotype network, Φst analyses indicated significant genetic structure among four geographical groups. Demographic analyses suggest that these populations have experienced a constant population expansion in the last 0.4–0.1 million of years. Discussion The weak, but significant, differences in Mediterranean and adjacent North-eastern Atlantic blue sharks revealed a complex phylogeographic structure, which appears to reject the assumption of panmixia across the study area, but also supports a certain degree of population connectivity across the Strait of Gibraltar, despite the lack of evidence of migratory movements observed by tagging data. Analyses of spatial genetic structure in relation to sex-ratio and size could indicate some level of sex/stage biased migratory behaviour