All is fish that comes to the net: metabarcoding for rapid fisheries catch assessment.

Monitoring marine resource exploitation is a key activity in fisheries science and biodiversity conservation. Since research surveys are time consuming and costly, fishery-dependent data (i.e., derived directly from fishing vessels) are increasingly credited with a key role in expanding the reach of ocean monitoring. Fishing vessels may be seen as widely ranging data-collecting platforms, which could act as a fleet of sentinels for monitoring marine life, in particular exploited stocks. Here, we investigate the possibility of assessing catch composition of single hauls carried out by trawlers by applying DNA metabarcoding to the dense water draining from fishing nets just after the end of hauling operations (hereafter "slush"). We assess the performance of this approach in portraying ß-diversity and examining the quantitative relationship between species abundances in the catch and DNA amount in the slush (read counts generated by amplicon sequencing). We demonstrate that the assemblages identified using DNA in the slush satisfactorily mirror those returned by visual inspection of net content (about 71% of species and 86% of families of fish) and detect a strong relationship between read counts and species abundances in the catch. We therefore argue that this approach could be upscaled to serve as a powerful source of information on the structure of demersal assemblages and the impact of fisheries.

Gonadal disorder in the thinlip grey mullet (Liza ramada, Risso 1827) as a biomarker of environmental stress in surface waters.

The aim of this study was to evaluate the use of gonadal alterations in the thinlip grey mullet (Liza ramada) as a biological indicator in assessing aquatic ecosystems health, with particular emphasis to river ecosystems exposed to sewage discharges. For this purpose, the reproductive status and the presence of gonadal alterations were studied in 206 mullets collected from two sites on the low course of the Tiber River, downstream of a large urban sewage treatment plant and in the estuarine area, and from an uncontaminated pond considered as reference site. Intersex and irregularly shaped gonads were observed in 20.8% of the mullets from the most polluted site, and intersex gonads in 10.3% of those from the estuarine area. No alterations were detected in the fish from the reference site, which also showed distinct stages of gonadal development. Conversely, unclear stages of testicular and ovary development were observed in the fish from the two polluted river sites. The results of this study suggest that L. ramada may represent a sentinel species in environmental risk assessment and support the use of gonadal alterations of this species as a bioindicator for extensive monitoring of pollution in lower stretches of rivers and estuarine areas.

VMSbase: an R-package for VMS and logbook data management and analysis in fisheries ecology.

VMSbase is an R package devised to manage, process and visualize information about fishing vessels activity (provided by the vessel monitoring system--VMS) and catches/landings (as reported in the logbooks). VMSbase is primarily conceived to be user-friendly; to this end, a suite of state-of-the-art analyses is accessible via a graphical interface. In addition, the package uses a database platform allowing large datasets to be stored, managed and processed vey efficiently. Methodologies include data cleaning, that is removal of redundant or evidently erroneous records, and data enhancing, that is interpolation and merging with external data sources. In particular, VMSbase is able to estimate sea bottom depth for single VMS pings using an on-line connection to the National Oceanic and Atmospheric Administration (NOAA) database. It also allows VMS pings to be assigned to whatever geographic partitioning has been selected by users. Standard analyses comprise: 1) métier identification (using a modified CLARA clustering approach on Logbook data or Artificial Neural Networks on VMS data); 2) linkage between VMS and Logbook records, with the former organized into fishing trips; 3) discrimination between steaming and fishing points; 4) computation of spatial effort with respect to user-selected grids; 5) calculation of standard fishing effort indicators within Data Collection Framework; 6) a variety of mapping tools, including an interface for Google viewer; 7) estimation of trawled area. Here we report a sample workflow for the accessory sample datasets (available with the package) in order to explore the potentialities of VMSbase. In addition, the results of some performance tests on two large datasets (1×10(5) and 1×10(6) VMS signals, respectively) are reported to inform about the time required for the analyses. The results, although merely illustrative, indicate that VMSbase can represent a step forward in extracting and enhancing information from VMS/logbook data for fisheries studies.

Skeletal anomaly monitoring in rainbow trout (Oncorhynchus mykiss, Walbaum 1792) reared under different conditions.

The incidence of skeletal anomalies could be used as an indicator of the "quality" of rearing conditions as these anomalies are thought to result from the inability of homeostatic mechanisms to compensate for environmentally-induced stress and/or altered genetic factors. Identification of rearing conditions that lower the rate of anomalies can be an important step toward profitable aquaculture as malformed market-size fish have to be discarded, thus reducing fish farmers' profits. In this study, the occurrence of skeletal anomalies in adult rainbow trout grown under intensive and organic conditions was monitored. As organic aquaculture animal production is in its early stages, organic broodstock is not available in sufficient quantities. Non-organic juveniles could, therefore, be used for on-growing purposes in organic aquaculture production cycle. Thus, the adult fish analysed in this study experienced intensive conditions during juvenile rearing. Significant differences in the pattern of anomalies were detected between organically and intensively-ongrown specimens, although the occurrence of severe, commercially important anomalies, affecting 2-12.5% of individuals, was comparable in the two systems. Thus, organic aquaculture needs to be improved in order to significantly reduce the incidence of severe anomalies in rainbow trout.

SMART: a spatially explicit bio-economic model for assessing and managing demersal fisheries, with an application to italian trawlers in the strait of sicily.

Management of catches, effort and exploitation pattern are considered the most effective measures to control fishing mortality and ultimately ensure productivity and sustainability of fisheries. Despite the growing concerns about the spatial dimension of fisheries, the distribution of resources and fishing effort in space is seldom considered in assessment and management processes. Here we propose SMART (Spatial MAnagement of demersal Resources for Trawl fisheries), a tool for assessing bio-economic feedback in different management scenarios. SMART combines information from different tasks gathered within the European Data Collection Framework on fisheries and is composed of: 1) spatial models of fishing effort, environmental characteristics and distribution of demersal resources; 2) an Artificial Neural Network which captures the relationships among these aspects in a spatially explicit way and uses them to predict resources abundances; 3) a deterministic module which analyzes the size structure of catches and the associated revenues, according to different spatially-based management scenarios. SMART is applied to demersal fishery in the Strait of Sicily, one of the most productive fisheries of the Mediterranean Sea. Three of the main target species are used as proxies for the whole range exploited by trawlers. After training, SMART is used to evaluate different management scenarios, including spatial closures, using a simulation approach that mimics the recent exploitation patterns. Results evidence good model performance, with a noteworthy coherence and reliability of outputs for the different components. Among others, the main finding is that a partial improvement in resource conditions can be achieved by means of nursery closures, even if the overall fishing effort in the area remains stable. Accordingly, a series of strategically designed areas of trawling closures could significantly improve the resource conditions of demersal fisheries in the Strait of Sicily, also supporting sustainable economic returns for fishermen if not applied simultaneously for different species.

Applications of self-organizing maps for ecomorphological investigations through early ontogeny of fish.

We propose a new graphical approach to the analysis of multi-temporal morphological and ecological data concerning the life history of fish, which can typically serves models in ecomorphological investigations because they often undergo significant ontogenetic changes. These changes can be very complex and difficult to describe, so that visualization, abstraction and interpretation of the underlying relationships are often impeded. Therefore, classic ecomorphological analyses of covariation between morphology and ecology, performed by means of multivariate techniques, may result in non-exhaustive models. The Self Organizing map (SOM) is a new, effective approach for pursuing this aim. In this paper, lateral outlines of larval stages of gilthead sea bream (Sparus aurata) and dusky grouper (Epinephelus marginatus) were recorded and broken down using by means of Elliptic Fourier Analysis (EFA). Gut contents of the same specimens were also collected and analyzed. Then, shape and trophic habits data were examined by SOM, which allows both a powerful visualization of shape changes and an easy comparison with trophic habit data, via their superimposition onto the trained SOM. Thus, the SOM provides a direct visual approach for matching morphological and ecological changes during fish ontogenesis. This method could be used as a tool to extract and investigate relationships between shape and other sinecological or environmental variables, which cannot be taken into account simultaneously using conventional statistical methods.

Testing species delimitations in four Italian sympatric leuciscine fishes in the Tiber River: a combined morphological and molecular approach.

Leuciscine fishes represent an important component of freshwater ichthyofauna endemic to northern Mediterranean areas. This lineage shows high intra-specific morphological variability and exhibits high levels of hybridization, two characteristics that contribute to systematic uncertainties, misclassification of taxa and, potentially, the mismanagement of biodiversity. This study focused on brook chub, Squalius lucumonis, an endemic taxon of Central Italy. The taxonomic status of this species has long been questioned, and a hybrid origin from sympatric leusciscines (S. squalus x Rutilus rubilio, or S. squalus x Telestes muticellus) has been hypothesised. A phenotypic (evaluating shape and meristic counts) and genetic (using mitochondrial and nuclear markers) investigation of these four taxa was conducted to test species delimitation in sympatric areas and to evaluate the taxonomic status of S. lucumonis. One hundred and forty-five individuals of all four taxa were collected within streams of the lowest portion of the Tiber River basin and analysed; this region encompasses a large portion of the S. lucumonis distribution. The different morphological and genetic approaches were individually examined, compared, and then combined in a quantitative model to both investigate the limits of each approach and to identify cases of misclassification. The results obtained confirm the cladogenetic non-hybrid origin of S. lucumonis, highlight the need for immediate conservation actions and emphasise the value of an integrated approach in the study of leuciscines evolution.

Environmental conditioning of skeletal anomalies typology and frequency in gilthead seabream (Sparus aurata L., 1758) juveniles.

In this paper, 981 reared juveniles of gilthead seabream (Sparus aurata) were analysed, 721 of which were from a commercial hatchery located in Northern Italy (Venice, Italy) and 260 from the Hellenic Center for Marine Research (Crete, Greece). These individuals were from 4 different egg batches, for a total of 10 different lots. Each egg batch was split into two lots after hatching, and reared with two different methodologies: intensive and semi-intensive. All fish were subjected to processing for skeletal anomaly and meristic count analysis. The aims involved: (1) quantitatively and qualitatively analyzing whether differences in skeletal elements arise between siblings and, if so, what they are; (2) investigating if any skeletal bone tissue/ossification is specifically affected by changing environmental rearing conditions; and (3) contributing to the identification of the best practices for gilthead seabream larval rearing in order to lower the deformity rates, without selections. The results obtained in this study highlighted that: i) in all the semi-intensive lots, the bones having intramembranous ossification showed a consistently lower incidence of anomalies; ii) the same clear pattern was not observed in the skeletal elements whose ossification process requires a cartilaginous precursor. It is thus possible to ameliorate the morphological quality (by reducing the incidence of severe skeletal anomalies and the variability in meristic counts of dermal bones) of reared seabream juveniles by lowering the stocking densities (maximum 16 larvae/L) and increasing the volume of the hatchery rearing tanks (minimum 40 m(3)). Feeding larvae with a wide variety of live (wild) preys seems further to improve juvenile skeletal quality. Additionally, analysis of the morphological quality of juveniles reared under two different semi-intensive conditions, Mesocosm and Large Volumes, highlighted a somewhat greater capacity of Large Volumes to significantly augment the gap with siblings reared in intensive (conventional) modality.

"Right" or "wrong"? Insights into the ecology of sidedness in european flounder, Platichthys flesus.

Sidedness polymorphism in flatfish has been linked to ecological selection between morphs. However, the alternate hypothesis that morphological differences between right- and left-sided forms may be due to errors during development, as a consequence of disturbed homeostasis, which still remains largely unexplored. Here, we examined the case of Platichthys flesus (flounder), a polymorphic flatfish exhibiting large and clinal variation in the frequency of the left-sided morph, which is the reversed condition in this generally right-sided species. An integrated approach consisting of the analyses of shape variation, stomach contents, and skeletal anomalies was used. Morphological differences were observed between morphs, which are in agreement with previous findings in a congeneric species (Platichthys stellatus). In parallel, significant differences in feeding choices were detected, suggesting a coherent association between subtle morphological differences between morphs and their use of trophic resources. Skeletal anomalies and meristic counts did not corroborate the hypothesis that morphometric divergence in reversed individuals may be caused or reinforced by developmental instability.

Lévy processes and stochastic von Bertalanffy models of growth, with application to fish population analysis.

The study of animal growth is a longstanding crucial topic of theoretical biology. In this paper we introduce a new class of stochastic growth models that enjoy two crucial properties: the growth path of an individual is monotonically increasing and the mean length at time t follows the classic von Bertalanffy model. Besides the theoretical development, the models are also tested against a large set of length-at-age data collected on Atlantic herring (Clupea harengus): the mean lengths and variances of the cohorts were directly estimated by least squares. The results show that the use of subordinators can lead to models enjoying interesting properties, in particular able to catch some specific features often observed in fish growth data. The use of subordinators seems to allow for an increased fidelity in the description of fish growth, whilst still conforming to the general parameters of the traditional von Bertalanffy equation.

Ecomorphology of morpho-functional relationships in the family of Sparidae: a quantitative statistic approach.

In many fish species, morphological similarity can be considered as a proxy for similarities in habitat use. The Sparidae family includes species that are recognized for common morphological features such as structure and positioning of the fins and specialized dentition. The aim of this study was to quantitatively describe the relationship of body shape morphology with habitat use, trophic level, and systematics in the majority of known Sparidae species (N = 92). This ecomorphological comparison was performed with a geometric morphometric approach considering as variables the Trophic Index (TROPH), the habitat (i.e., classified as demersal, benthopelagic and reef associated) and the phylogenetic relationship of species at the subfamily level. The analysis by the TROPH variable showed a positive relation with shape because the morphological features of all the species are strongly correlated with their trophic behavior (e.g., herbivore species have a smaller mouth gap that make them able to feed upon sessile resources). The morphological analysis according to the Habitat variable was used to classify species according to a feeding-habitat niche in terms of portion of the water column and seabed space where species mostly perform their behavioral activities. We described three kinds of morphological designs in relation to a benthopelagic, demersal and reef-associated habit. The six subfamily groups were morphologically well distinguishable and the cladogram relative to Mahalanobis' morphological distances was compared with those proposed by other authors. We also quantified the phylogenetic relationship among the different subfamilies based on the analysis of shape in relation to trophic ecology, confirming the observations of the authors.

Shape and size variation: growth and development of the dusky grouper (Epinephelus marginatus Lowe, 1834).

Demersal fishes have complex life cycles that involve an ontogenetic change in morphology, physiology, and behavior, as their pelagic larval stages colonize benthic habitats. The developmental transition between larvae and juveniles leads to very complex processes of morphogenesis and differentiation. These processes primarily determine changes in external morphology, which is shaped by selective pressures to optimize performance for basic activities such as swimming, escape from predators, and feeding. Fishes have provided fertile grounds for ecomorphological investigations throughout ontogeny, as the role of changing morphology in inducing ontogenetic niche shifts is not always clear. In this framework, some studies have demonstrated that certain species undergo gradual changes, whereas other species experience threshold effects in their ecomorphological relationships during ontogeny. In this study, the intraspecific allometry of the dusky grouper was examined. Geometric morphometric tools were used to quantify shape changes through the development, and a modular approach was also applied to analyze the pattern of covariation between three distinct blocks (head, trunk, and tail). For this purpose, a two-block Partial Least Square was computed. This method reveals that the pattern of changes in the overall body shape is the result of the modularized changes of these blocks.

Light and shape: a contribution to demonstrate morphological differences in diurnal and nocturnal teleosts.

Light intensity is an important environmental factor affecting the structure of fish assemblages during the day-night cycle. Light influences how organisms perceive their environment, modulating their intraspecific and interspecific relationships. The relationship between light intensity variations and biological cycles should be observed at the level of organismal morphology. In this study the relationship between activity rhythms, thus light intensity experienced by fish in the period of major activity and external morphology, have been investigated. The morphological traits of 97 selected fish species were compared in order to determine the existence of a common morphological plan in agreement with their diurnal or nocturnal activity rhythm. Species sorting was performed by maximizing the diversity of activity rhythm, habitat choice, ecology, and trophic habits within the same family, to assess the importance of the day-night cycle on species morphology in relation to other environmental features. The morphological characters selected for the geometric morphometric analysis were body profile and the position of mouth, eye, pelvic, pectoral, dorsal, and caudal fin. The present analysis allowed different consensus forms for nocturnal and for diurnal species to be identified. Two-block Partial Least Squares analysis was then performed for the purpose of modeling the covariation between the form and two important external variables (ecology and activity).

[Fish-based assessment methods for the ecological status of aquatic systems]. / I pesci nella valutazione dello stato ecologico.

A short overview of fish-based assessment methods for aquatic systems is presented. Multimetric indices, as, e.g., the index of biotic integrity (IBI), firstly developed in USA and later adapted for European river basins and other countries, are shortly described. Non-multimetric indices are also discussed, e.g. the ichthyological index (II) and the index of ecological status of fish communities (ISECI), both proposed for monitoring Italian rivers. Moreover, statistical and predicting methods based on machine learning techniques are described. Finally, a new approach for developing standardised fish-based methods useful to assess ecological status of Italian rivers is proposed. Although rather complex, the use of bony fish in biomonitoring is promising and requires a multidisciplinary approach to be adopted.

Fatty acid pattern, oxidation product development, and antioxidant loss in muscle tissue of rainbow trout and Dicentrarchus labrax during growth.

The levels of hydrophilic, lipophilic, and enzymatic antioxidants, the oxidative damage to lipids and proteins, and the fatty acid patterns of triglyceride and phospholipid fractions were assayed in fresh muscle tissue of rainbow trouts (Oncorhynchus mykiss) and sea basses (Dicentrarchus labrax) during aging, to investigate the correlation between oxidative stress and aging processes in fish. The present studies suggests that lipid peroxidation and accumulation of oxidized proteins during in vivo aging are most likely to be linked with an age-dependent decline of lipophilic antioxidants (CoQH(2), CoQ, and vitamin E) and vitamin C contents in muscle tissue, whereas fish aging is not linked to a decline in antioxidant enzymes and reduced glutathione levels. Lipophilic antioxidant and vitamin C levels represent a reliable marker of oxidative stress during aging, and their determination might be useful for the assessment of fish age.

Fatty acid composition and antioxidant levels in muscle tissue of different Mediterranean marine species of fish and shellfish.

The levels of hydrophilic, lipophilic, and enzymatic antioxidants, as well as the fatty acids composition, of triglyceride and phospholipid fractions were determined in the muscle tissue of 21 species of teleosts, 3 species of cephalopods, and 6 species of crustaceans, just caught from the central Tyrrhenian Sea (Mediterranean Sea). The enzymatic activities and the levels of low-molecular-weight antioxidants, and the percentages of fatty acids, showed marked interspecies differences. Our results showed that total polyunsaturated fatty acids (21.7-61.5%) were the highest, followed by saturated (16.9-41.3%) and monounsaturated (9.1-42.8%) fatty acids. The total n-3 fatty acids content (16.6-57.1%) was found to be higher than the total n-6 fatty acids content (4.1-10.6%). All of the species studied had an n-3/n-6 ratio of more than 1, confirming the great importance of fish and shellfish as a significant dietary source of n-3 polyunsaturated fatty acids and their beneficial role in the Mediterranean type of diet.