Corrado Piccinetti

Long-term changes in demersal resources of the Adriatic Sea: comparison between trawl surveys carried out in 1948 and 1998

Abstract Trawl surveys carried out in the Adriatic Sea in 1948 and 1998 provided information on the changes that occurred in the composition and distribution of demersal fish resources over the 50 years period. The main change was the decrease of elasmobranchs diversity and frequency, mostly referring to the Hypotremata group. Life history parameters appeared to be determining factors, since small sized species such as the smallspotted catshark ( S. canicula ) or the brown ray ( R. miraletus ) were frequently collected in both surveys, while bigger shark species and most other rays disappeared or were rarely found during the 1998 survey. Regarding bony fishes, common species were collected with similar frequency in both surveys; some fishery target species (e.g. the hake) have similar percentage values (biomass) in both surveys, while others (the silver scabbardfish) have increased values, and some (mainly small sized species such as red mullet) replace elasmobranchs at the top of the catches. Despite the low number of observations (two surveys only) and some sampling differences, the data provide an interesting long-term comparison.

Unite research with what citizens do for fun: “recreational monitoring” of marine biodiversity

Institutes often lack funds and manpower to perform large-scale biodiversity monitoring. Citizens can be involved, contributing to the collection of data, thus decreasing costs. Underwater research requires specialist skills and SCUBA certification, and it can be difficult to involve volunteers. The aim of this study was to involve large numbers of recreational divers in marine biodiversity monitoring for increasing the environmental education of the public and collecting data on the status of marine biodiversity. Here we show that thousands of recreational divers can be enrolled in a short time. Using specially formulated questionnaires, nonspecialist volunteers reported the presence of 61 marine taxa encountered during recreational dives, performed as regular sport dives. Validation trials were carried out to assess the accuracy and consistency of volunteer-recorded data, and these were compared to reference data collected by an experienced researcher. In the majority of trials (76%) volunteers performed with an accuracy and consistency of 50-80%, comparable to the performance of conservation volunteer divers on precise transects in other projects. The recruitment of recreational divers involved the main diving and tour operators in Italy, a popular scientific magazine, and mass media. During the four-year study, 3825 divers completed 18757 questionnaires, corresponding to 13539 diving hours. The volunteer-sightings-based index showed that in the monitored area the biodiversity status did not change significantly within the project time scale, but there was a significant negative correlation with latitude, suggesting improved quality in the southernmost areas. This trend could be related to the presence of stressors in the northern areas and has been supported by investigations performed by the Italian Ministry of the Environment. The greatest limitation with using volunteers to collect data was the uneven spatial distribution of samples. The benefits were the considerable amounts of data collected over short time periods and at low costs. The successful development of citizen-based monitoring programs requires open-mindedness in the academic community; advantages of citizen involvement in research are not only adding large data sets to the ecological knowledge base but also aiding in the environmental education of the public.

Mitochondrial DNA Sequence Variation Suggests the Lack of Genetic Heterogeneity in the Adriatic and Ionian Stocks of Sardina pilchardus

Abstract: A genetic stock structure analysis of 11 sardine samples from the Adriatic Sea and Ionian neighboring area was carried out through sequence variation analysis of a 307-bp cytochrome b gene fragment in order to identify self-recruiting units in the Adriatic Sardina pilchardus stock. The overall lack of genetic subdivision among samples detected by analysis of molecular variance, pairwise Φst values, and the exact test of population differentiation indicates this sardine stock is part of a larger self-recruiting population whose boundaries are larger than the investigated area. This conclusion is in agreement with preliminary allozymic and mitochondrial DNA restriction fragment length polymorphism data, but contradicts the previous identification of 2 subpopulations of sardines in the Adriatic Sea argued on morphologic differences, which could be rather attributed to different hydrographic or ecologic conditions occurring in different areas of the Adriatic Sea. The reduced gene flow observed between Adriatic-Ionian and Spanish sardine geographic samples (P < 0.001) suggests that reproductively isolated populations of sardines may occur in the Mediterranean Sea.

Microsatellite DNA variation reveals high gene flow and panmictic populations in the Adriatic shared stocks of the European squid and cuttlefish (Cephalopoda)

In the semienclosed Adriatic Sea, the shared stocks of the cephalopods Loligo vulgaris and Sepia officinalis represent important marine fisheries resources exploited by all coastal countries. The improving of knowledge on the demographic features of these shared stocks is internationally relevant for adopting responsible management and conservation of these marine resources. Analyses of microsatellite variation in geographical samples collected from all parts of the Adriatic Sea were performed using arrays of species-specific di-nucleotide and tri-nucleotide loci. In L. vulgaris the level of genetic variability was consistent with that observed in other loliginid species, whereas the S. officinalis stock showed a microsatellite variation markedly lower than that estimated for the Atlantic and Mediterranean populations collected around the Iberian peninsula. The weak spatial genetic differentiation, the discordant results of the genetic divergence estimators and the lack of any geographical cline in the spatial genetic differences suggest the occurrence of single genetically homogeneous populations within the Adriatic stocks of both species, recommending a coordinated management of the squid and cuttlefish by the Adriatic fishing countries. On the contrary, significant differences detected in temporal replicates of S. officinalis might suggest that allelic frequency can change relating to reproductive behaviour.

Development of molecular and morphological markers to improve species-specific monitoring and systematics of Northeast Atlantic and Mediterranean skates (Rajiformes)

The Northeast Atlantic and Mediterranean skates (Rajidae) showed remarkable species diversity but with high morphological and ecological conservatism. Since skates are particularly vulnerable to the bottom trawl fishery, species-specific demographic surveys as well as studies defining life history and evolutionary traits are important in prioritising conservation programs. However, the identification of juveniles and adults of some species may be difficult using referenced guidelines and identification keys. Therefore, we attempt to develop markers for species identification through the parallel analysis of a 16S rDNA gene sequence and of several morphological characters on 135 individuals collected by trawl surveys in the Adriatic Sea and putatively assigned to six taxa. Species-specific haplotypes were defined for Raja miraletus, Raja montagui, Dipturus oxyrinchus, since a solid accordance between species boundaries and well-differentiated haplotypes was observed. Comparative analysis of 16S rDNA sequences allowed the identification of three juvenile specimens of Leucoraja circularis, a species that rarely occurs in the Adriatic Sea. On the contrary, morphological traits and haplotype distribution were largely discordant in Raja asterias and R. clavata. While all putative R. clavata individuals showed a unique haplotype (H-CLA), only 8 of 30 putative R. asterias individuals possessed a second weakly divergent haplotype (H-AST). The remaining 22 R. asterias carried the H-CLA. The multivariate analyses of morphometric and meristic characters in putative R. clavata and R. asterias revealed the clustering of individuals regardless of haplotypes. However, a bimodal distribution of R. asterias and R. clavata samples would suggest that two separated taxa might exist, both sharing the two 16S rDNA haplotypes. The haplotype distribution appeared to be significantly correlated only to the standardised disc length/total length (DL/TL) variation. Three alternative explanations may support this scenario: (i) an incomplete lineage sorting process in two morphologically yet distinct taxa; (ii) a recent hybridisation between the two taxa; (iii) the two taxa are morphologically plastic species and all considered morphological characters may be misleading in discriminating between them at all maturity stages, except for the DL/TL. However, further analyses on larger data sets and using molecular key markers (i.e. nuclear genes) will be needed to definitely resolve the status of these taxa. Molecular relationships among rajid taxa are largely consistent with systematics based on internal and external anatomical features. This multidisciplinary study contributed to defining the pattern of species diversity and abundance of rajids in the Adriatic Sea.

First Microsatellite Loci of Red Mullet (Mullus barbatus) and Their Application to Genetic Structure Analysis of Adriatic Shared Stock

In order to study the genetic structure of the Adriatic shared stock of red mullet (Mullus barbatus), we developed a set of dinucleotide microsatellite markers. A dinucleotide-enriched genomic library was obtained, and 6 polymorphic dinucleotide loci were successfully optimized. The markers showed high expected heterozygosity (from 0.68 to 0.92) and allele number (from 12 to 33); thus they appear to be suitable for detecting genetic differences in the population of red mullet. Four Adriatic samples were subsequently analyzed for microsatellite variation, and the results showed subtle but statistically significant genetic differentiation, indicating that the Adriatic red mullet may group into local, genetically isolated populations. No correlation between geographic distance and genetic differentiation was observed. In addition, the evidence of recent bottlenecks in the Adriatic samples indicates that the observed population subdivision might reflect random local allelic variations, generated by reproductive success, survival rates, or fishing pressure.

Comparative Analysis of a Mitochondrial DNA Control Region Fragment Amplified from Three Adriatic Flatfish Species and Molecular Phylogenesis of Pleuronectiformes

Abstract The 5′-end of the mitochondrial control region of three Pleuronectiformes from the Adriatic Sea, Platichthys flesus italicus (Adriatic flounder), Solea vulgaris (common sole), and Solea kleini (Kleins sole), was sequenced and compared with that of six other flatfish species from the families Pleuronectidae and Bothidae. The sequence structures of all flatfishes appear very similar and consist of alternate short segments with low, medium, and high rates of nucleotide substitution. Four conserved 19-bp repeats occur at the beginning of the European and Adriatic flounder sequences. The common occurrence of tandem arrays in fish control regions could be related to a stable secondary structure. Molecular phylogenetic relationships among Pleuronectiformes agree well with previous morphologic data at all taxonomic levels. Molecular analyses could therefore contribute to resolving phylogenetic and taxonomic debates within the Pleuronectiformes.

Mitochondrial DNA variation, phylogenetic relationships, and evolution of four mediterranean genera of soles (soleidae, pleuronectiformes).

To increase knowledge about the systematics and evolution of Mediterranean soles, we assessed mitochondrial DNA variation, molecular phylogeny, and evolution in eight species from the genera Solea, Microchirus, Monochirus, and Buglossidium by large ribosomal subunit (16S) and cytochrome b (cytb) sequence analysis. Relevant molecular features are the great variation of base composition among species at the third codon in cytb and the heterogeneity of the nucleotide substitution rate. Phylogenies recovered using 16S nucleotide and cytb amino acid sequences agree with those based on morphology in assessing monophyly of Solea species and ancestry of Buglossidium luteum, but they are against the intergeneric differentiation of Microchirus and Monochirus. Conversely, phylogenetic trees based on cytb nucleotide sequences yielded relationships among taxa regardless of their evolutionary histories. The incongruities between morphological and molecular issues suggest the need for reassessing the systematic value of some morphological characters. Approximate estimates of the divergence time of Mediterranean soleid lineages range from 40 to 13 Mya (Oligocene–Miocene), indicating an ancient origin for the group.

Determination of Lmax for Atlantic Bluefin Tuna, Thunnus thynnus (L.), from Meta-Analysis of Published and Available Biometric Data

A meta-analysis of the straight fork lengths (herewith abbreviated as L) of 2,458,028 Atlantic bluefin tuna, Thunnus thynnus (L.), taken from 224 scientific publications and unpublished L data from scientific organizations and fishing companies spanning most of the known Atlantic and Mediterranean Atlantic bluefin tuna fisheries dating from 1605 to 2011, give L values ranging from L min = 20 cm and L max = 330 cm. The results indicate that the parameter L ∞ = 318.85 cm of the growth equation used by ICCATs Standing Committee on Research and Statistics Atlantic bluefin tuna assessment group for the eastern stock (Lt = 318.85 [1 – e−0.093 (t + 0.97)]) lies within the confidence limits of the maximum Ls presented in the study: L max = 319.93 ± 11.3 cm, confirming that this equation perfectly fits the biology of the growth of this species. These conclusions are also valid for the equation for the western stock (Lt = 314.90 [1 – e−0.089 (t +1.13)]). The ICCAT Atlantic bluefin tuna database contains numerous records of Atlantic bluefin tuna L outside the biological feasibility, and solutions are provided to recognize and remove these outliers based on the application of fixed values of Fultons condition factor (K) between 1.4 and 2.6 and appropriate L-W relationships to correct this situation in the future.

Participating in a Citizen Science Monitoring Program: Implications for Environmental Education

Tourism is of growing economical importance to many nations, in particular for developing countries. Although tourism is an important economic vehicle for the host country, its continued growth has led to on-going concerns about its environmental sustainability. Coastal and marine tourism can directly affect the environment through direct and indirect tourist activities. For these reasons tourism sector needs practical actions of sustainability. Several studies have shown how education minimizes the impact on and is proactive for, preserving the natural resources. This paper evaluates the effectiveness of a citizen science program to improve the environmental education of the volunteers, by means of questionnaires provided to participants to a volunteer-based Red Sea coral reef monitoring program (STEproject). Fifteen multiple-choice questions evaluated the level of knowledge on the basic coral reef biology and ecology and the awareness on the impact of human behaviour on the environment. Volunteers filled in questionnaires twice, once at the beginning, before being involved in the project and again at the end of their stay, after several days participation in the program. We found that the participation in STEproject significantly increased both the knowledge of coral reef biology and ecology and the awareness of human behavioural impacts on the environment, but was more effective on the former. We also detected that tourists with a higher education level have a higher initial level of environmental education than less educated people and that the project was more effective on divers than snorkelers. This study has emphasized that citizen science projects have an important and effective educational value and has suggested that tourism and diving stakeholders should increase their commitment and efforts to these programs