Co-inertia multivariate approach for the evaluation of anthropogenic impact on two commercial fish along Tyrrhenian coasts.

Aliphatic hydrocarbon levels were determined by the GC/MS technique in fish livers of Engraulis encrasicolus (Ee) and Trachurus trachurus (Tt), collected from a particular area of the Mediterranean Sea, called GSA 10, which is located exactly in Tyrrhenian Sea between Campania coast and North Sicily coast. The aim was to evaluate their potential use as specific bioindicators towards this class of contaminants. Both Tt and Ee are considered to be pollution monitoring bioindicators, due to their dominance in marine communities and economic fishing interest. Ee showed a higher tendency to bioaccumulate TAHs, due to the lower quantity of fatty acids in liver tissues with respect to Tt. The area under study has been characterised a) chemically with the acquisition of temperature, oxygen and salinity profiles along the water column, and b) ecologically with the determination of amino acid contents in fish eyes, in order to gain information on the adaptation to environmental changes. Moreover, specific activities of two hydrolytic enzymes, such as alkaline phosphatase and peroxidase in fish epidermal mucus, together with lactate in blood plasma and cortisol levels, have been investigated for the first time, in order to obtain insights into the effects of hydrocarbons on animal welfare. A multiple co-inertia analysis was also applied to chemical and environmental parameters, in order to explore any possible correlation between different variables. The multivariate approach showed a clear spatial distribution between environmental and chemical variables in Ee, whilst there was an absence of a spatial trend in Tt. Moreover, the chemometric analysis showed a very high correlation between amino acid profiles and environmental variables for both species, confirming the possibility of being used as ecological welfare indices for short-term environmental variations.

Underwater vocal complexity of Arctic seal Erignathus barbatus in Kongsfjorden (Svalbard).

In this study the description of underwater vocal repertoire of bearded seal in Svalbard (Norway) was extended. Two autonomous passive acoustic recorders were deployed for one year (August 2014-July 2015) in the inner and outer parts of the Kongsfjorden, and 1728 h were recorded and 17 220 vocalizations were found. Nine different vocalization classes were identified and characterized using ten acoustic parameters. The calls showed heterogeneous spectral features, but share the descending trend of frequency modulation. The different classes emerged were discriminated primarily by bandwidth and duration, and then by minimum frequency, central frequency, and maximum frequency in this order. This study represents a step forward to improve the understanding of the acoustic behaviour and the social function of these calls, and identified long passive acoustic monitoring as an effective method to assess vocal complexity and the ecology of marine species producing sounds.

Distribution of Cd and As in organs and tissues of four marine mammal species stranded along the Italian coasts.

Concentrations of Cd and As were determined in organs and tissues (muscle, heart, kidney, lung and liver) of four cetacean species (Stenella coeruleoalba, Truncatus truncatus, Grampus griseus and Ziphius cavirostris) stranded along the Italian coasts during the period 2000-2009. Significant differences were found between Cd concentrations in the different analysed tissues. Particularly, the kidney shows the highest concentrations of Cd in all analysed specimens, followed by the liver. No systematic or statistically significant difference in As concentrations was found in the analysed tissues. Considerable differences of Cd and As values in samples from different geographical areas (Sicily Channel, Adriatic Sea and Tyrrhenian Sea) were found, with the highest values measured in samples from the Sicily Channel. This basically suggests that anthropogenic and natural sources significantly affect Cd and As content in cetaceans.

New Evaluation of Postovulatory Follicle Degeneration at High-Temperature Regimes Refines Criteria for the Identification of Spawning Cohorts in the European Anchovy (Engraulis encrasicolus).

Accurate stock assessment estimates of fish resources are essential in fishery management. Wide fluctuations in abundance arising from variations in reproductive success are characteristic of many marine fish populations, including multiple spawner species. The proportion of females spawning per day is crucial in the application of egg production methods for spawning biomass evaluation and, usually, is assessed by postovulatory follicle (POF) method. Describing each degeneration stage of POF based upon its histological features allows for obtaining an aging key for postovulatory follicles. The commercially valuable European anchovy (Engraulis encrasicolus) was selected as a case study, which breeds during the summer in temperate waters (24 °C-25 °C). A collection of ovary slides, sampled in the central Mediterranean Sea during the spawning peak, provided a 24 h cycle coverage. These observations allowed us to evaluate the duration of each POF stage at water temperature experienced by anchovy in the study area. Present results demonstrated the POF degeneration progress at a faster rate than reported by previous investigations, carried out in cooler oceanic waters. Furthermore, the present study displayed the presence of two anchovy spawning cohorts sampled along a 24-h cycle. Therefore, this study not only provides useful insight for more accurate POF degeneration evaluation in temperate waters, but also suggests that current estimates should be complemented with validation studies according to different temperature regimes.

Growth-related trophic changes of Thunnus thynnus as evidenced by stable nitrogen isotopic values in the first dorsal spine.

The bluefin tuna, Thunnus thynnus, is a highly migratory and long-living fish at the top of the pelagic food web. As top predator, it plays a key role in the stability of marine food webs by exerting top-down control on its prey. The diet composition of bluefin tuna varies in relation to its growth, seasons and migratory patterns, making it difficult to evaluate spatial and temporal effects. This latter aspect is further complicated to be determined during the first months of life, when T. thynnus specimens have a rapid growth rate leading to changes in the trophic status. In this study, the potential collagen-related effects on δ15N and δ13C values were evaluated on the whole spine of adult tuna specimens collected in the central Mediterranean Sea. Obtained results showed non-significant differences between extracted and non-extracted collagen samples for δ15N in whole spine, allowing adopting the isotopic analysis both for annuli in the spine section of adults and for younger specimens, whose spine size does not permit the collagen extraction. Specifically, isotopic analysis of whole spine of the young of the year specimens, showed a rapid change in δ15N values with length, following an exponential model. For older specimens, δ15N values were higher and varied around a plateau, likely due to a higher specificity in the choice of prey and/or to change in the geographical location. Such variability was also mirrored in annuli of spines sections of adult tunas. As far as δ13C values are concerned, a strong collagen-related effect was evidenced, likely highlighting the influence of lipids. Consequently, δ13C analysis may be used only on adult specimens where collagen extraction is possible. This research also showed how isotopic analysis of both whole sample and sequence of annuli in the cross-section of dorsal spine might produce isotopic profiles useful to detect specific trophic dynamics along the bluefin tuna growth.

First annulus formation in the European anchovy; a two-stage approach for robust validation.

The age determination in fast-growing short-living species, such as European anchovy (Engraulis encrasicolus), has been widely recognized as a difficult task and bias introduced by readers leads to bias in reconstructing the population age structure. In this context, it is worth to note that age structure of fish population represents key information in fishery ecology and for stock assessment models. The uncertainty in estimating the age of the European anchovy by otolith reading is linked to the number of false-growth increments (checks) laid down before the annulus formation. While direct validation methods (e.g. mark-recapture, rearing, radiochemical dating) are difficult to implement specially for this short living species, the use of different indirect methods, supported by a coherent statistical approach, represents a robust and easier validation tool. A statistical modeling approach has been here adopted to assess the coherence of two well-known methods, namely Edge Analysis and Marginal Increment Analysis, in order to validate the first annulus formation in European anchovy. Both methodologies in two different yearly cycles converged toward the same result, thus confirming the annulus identification for the first year class. In addition, the completion dates of the checks and the first annulus were computed in order to gain a better insight into otolith growth dynamic. According to the species spawning period, the completion date of the first annulus falls in the summer period, while the first and second checks completion dates were mostly found in summer and winter respectively. General additive models using marginal increments as dependent variable showed a significant effect of the month, highlighting the presence of only one clear minimum in July/August, as well as specific relationships with condition factor and gonadosomatic index. Modeling the otolith edge morphology, the probability to find a hyaline band displayed in both years a similar shape, characterized by a minimum in July/August and higher values between November and January. The obtained results evidenced temporally coherent patterns providing a better insight in the otolith growth dynamic as well as a more robust validation of the first annulus formation in the European anchovy.

Condition of pteropod shells near a volcanic CO2 vent region.

Natural gradients of pH in the ocean are useful analogues for studying the projected impacts of Ocean Acidification (OA) on marine ecosystems. Here we document the in situ impact of submarine CO2 volcanic emissions (CO2 vents) on live shelled-pteropods (planktonic gastropods) species Creseis conica in the Gulf of Naples (Tyrrhenian Sea, Mediterranean). Since the currents inside the Gulf will likely drive those pelagic calcifying organisms into and out of the CO2 vent zones, we assume that pteropods will be occasionally exposed to the vents during their life cycle. Shell degradation and biomass were investigated in the stations located within and nearby the CO2 vent emission in relation to the variability of sea water carbonate chemistry. A relative decrease in shell biomass (22%), increase in incidence of shell fractures (38%) and extent of dissolution were observed in Creseis conica collected in the Gulf of Naples compared to those from the Northern Tyrrhenian Sea (control stations). These results suggest that discontinuous but recurrent exposure to highly variable carbonate chemistry could consistently affect the characteristic of the pteropod shells.

Liver melanomacrophage centres and CYP1A expression as response biomarkers to environmental pollution in European anchovy (Engraulis encrasicolus) from the western Mediterranean Sea.

The goal of the present study was to verify the suitability of using melanomacrophage centres (MMCs) as response biomarkers of marine pollution in European anchovy, which are short-lived, migratory, small pelagic fish. This suitability was verified by analysing the MMC density and cytochrome P450 monooxygenase 1A (CYP1A) expression in livers of anchovies from four areas of southern Italy. Age 2 anchovies sampled from three areas exposed to pollutants of industrial/agricultural origin (Gulf of Gela, Mazara del Vallo and Gulf of Naples) showed liver areas occupied by MMCs and numbers of MMCs that were significantly higher than those in the anchovies from Pozzallo, which is a marine area not subjected to any source of pollution. Anti-CYP1A immunoreactivity was observed in the hepatocytes of all specimens sampled from the Gulf of Gela. These findings suggest the utility of liver MMCs as biomarkers of exposure to pollutants in this small pelagic fish.

Spatio-temporal dynamics of a planktonic system and chlorophyll distribution in a 2D spatial domain: matching model and data.

Field data on chlorophyll distribution are investigated in a two-dimensional spatial domain of the Mediterranean Sea by using for phytoplankton abundances an advection-diffusion-reaction model, which includes real values for physical and biological variables. The study exploits indeed hydrological and nutrients data acquired in situ, and includes intraspecific competition for limiting factors, i.e. light intensity and phosphate concentration. As a result, the model allows to analyze how both the velocity field of marine currents and the two components of turbulent diffusivity affect the spatial distributions of phytoplankton abundances in the Modified Atlantic Water, the upper layer of the water column of the Mediterranean Sea. Specifically, the spatio-temporal dynamics of four phytoplankton populations, responsible for about 80% of the total chlorophyll a, are reproduced. Results for phytoplankton abundances obtained by the model are converted in chlorophyll a concentrations and compared with field data collected in twelve marine sites along the Cape Passero (Sicily)- Misurata (Libya) transect. Statistical checks indicate a good agreement between theoretical and experimental distributions of chlorophyll concentration. The study can be extended to predict the spatio-temporal behaviour of the primary production, and to prevent the consequent decline of some fish species in the Mediterranean Sea.

Insights on the drivers of genetic divergence in the European anchovy.

Anchovies represent the largest world's marine fish catches and the current threats on their populations impose a sustainable exploitment based on sound scientific information. In the European anchovy (Engraulis encrasicolus), the existence of several populations has been proposed but a global view is missing. Using a multidisciplinary approach, here we assessed the divergence among different ecotypes and its possible causes. SNPs have revealed two functionally distinct ecotypes overlapping in the Central Mediterranean, with one ecotype confined near the river estuaries. The same SNPs outliers also segregated two distinct populations in the near Atlantic, despite their large spatial distance. In addition, while most studies suggested that adaptation to low salinity is key to divergence, here we show that the offshore ecotype has higher environmental tolerance and an opportunistic feeding behaviour, as assessed by the study of environmental conditions, anchovy diet and trophic levels, and passive egg dispersal. These results provide insights into the anchovy evolutionary history, stressing the importance of behaviour in shaping ecotypes.

Amino acid composition in eyes from zebrafish (Danio rerio) and sardine (Sardina pilchardus) at the larval stage.

A comparative study was performed to identify differences in the amino acid composition of the eyes from zebrafish (Danio rerio) and sardine (Sardina pilchardus) larvae and their link to the environmental adaption of the species. Amino acids in the acidic hydrolysates of eyes from 11 zebrafish and 12 sardine were determined with the use of high-performance liquid chromatography involving precolumn derivatization with ortho-phthalaldehyde. Differences in the content of most amino acids were detected between zebrafish and sardine. These amino acids were aspartate, glutamate, serine, glycine, threonine, arginine, methionine, valine, phenylalanine, isoleucine, leucine and lysine. Of particular note, the percentage of methionine in zebrafish eyes was much higher than that in sardine, whereas the opposite was observed for glutamate and glycine. These results indicate that zebrafish and sardine likely have experienced differences in adaptation to environmental changes. We suggest that the amino acid composition of eyes represents a powerful tool to discriminate between species characterized by different lifestyle and inhabiting different environments.

Biocomplexity in Populations of European Anchovy in the Adriatic Sea.

The sustained exploitation of marine populations requires an understanding of a species' adaptive seascape so that populations can track environmental changes from short- and long-term climate cycles and from human development. The analysis of the distributions of genetic markers among populations, together with correlates of life-history and environmental variability, can provide insights into the extent of adaptive variation. Here, we examined genetic variability among populations of mature European anchovies (n = 531) in the Adriatic (13 samples) and Tyrrhenian seas (2 samples) with neutral and putative non-neutral microsatellite loci. These genetic markers failed to confirm the occurrence of two anchovy species in the Adriatic Sea, as previously postulated. However, we found fine-scale population structure in the Adriatic, especially in northern areas, that was associated with four of the 13 environmental variables tested. Geographic gradients in sea temperature, salinity and dissolved oxygen appear to drive adaptive differences in spawning time and early larval development among populations. Resolving adaptive seascapes in Adriatic anchovies provides a means to understand mechanisms underpinning local adaptation and a basis for optimizing exploitation strategies for sustainable harvests.

Interannual Changes in Biomass Affect the Spatial Aggregations of Anchovy and Sardine as Evidenced by Geostatistical and Spatial Indicators.

Geostatistical techniques were applied and a series of spatial indicators were calculated (occupation, aggregation, location, dispersion, spatial autocorrelation and overlap) to characterize the spatial distributions of European anchovy and sardine during summer. Two ecosystems were compared for this purpose, both located in the Mediterranean Sea: the Strait of Sicily (upwelling area) and the North Aegean Sea (continental shelf area, influenced by freshwater). Although the biomass of anchovy and sardine presented high interannual variability in both areas, the location of the centres of gravity and the main spatial patches of their populations were very similar between years. The size of the patches representing the dominant part of the abundance (80%) was mostly ecosystem- and species-specific. Occupation (area of presence) appears to be shaped by the extent of suitable habitats in each ecosystem whereas aggregation patterns (how the populations are distributed within the area of presence) were species-specific and related to levels of population biomass. In the upwelling area, both species showed consistently higher occupation values compared to the continental shelf area. Certain characteristics of the spatial distribution of sardine (e.g. spreading area, overlapping with anchovy) differed substantially between the two ecosystems. Principal component analysis of geostatistical and spatial indicators revealed that biomass was significantly related to a suite of, rather than single, spatial indicators. At the spatial scale of our study, strong correlations emerged between biomass and the first principal component axis with highly positive loadings for occupation, aggregation and patchiness, independently of species and ecosystem. Overlapping between anchovy and sardine increased with the increase of sardine biomass but decreased with the increase of anchovy. This contrasting pattern was attributed to the location of the respective major patches combined with the specific occupation patterns of the two species. The potential use of spatial indices as auxiliary stock monitoring indicators is discussed.

Habitat selection response of small pelagic fish in different environments. Two examples from the oligotrophic Mediterranean Sea.

A number of scientific papers in the last few years singled out the influence of environmental conditions on the spatial distribution of fish species, highlighting the need for the fisheries scientific community to investigate, besides biomass estimates, also the habitat selection of commercially important fish species. The Mediterranean Sea, although generally oligotrophic, is characterized by high habitat variability and represents an ideal study area to investigate the adaptive behavior of small pelagics under different environmental conditions. In this study the habitat selection of European anchovy Engraulis encrasicolus and European sardine Sardina pilchardus is analyzed in two areas of the Mediterranean Sea that largely differentiate in terms of environmental regimes: the Strait of Sicily and the North Aegean Sea. A number of environmental parameters were used to investigate factors influencing anchovy and sardine habitat selection. Acoustic surveys data, collected during the summer period 2002-2010, were used for this purpose. The quotient analysis was used to identify the association between high density values and environmental variables; it was applied to the entire dataset in each area in order to identify similarities or differences in the "mean" spatial behavioral pattern for each species. Principal component analysis was applied to selected environmental variables in order to identify those environmental regimes which drive each of the two ecosystems. The analysis revealed the effect of food availability along with bottom depth selection on the spatial distribution of both species. Furthermore PCA results highlighted that observed selectivity for shallower waters is mainly associated to specific environmental processes that locally increase productivity. The common trends in habitat selection of the two species, as observed in the two regions although they present marked differences in hydrodynamics, seem to be driven by the oligotrophic character of the study areas, highlighting the role of areas where the local environmental regimes meet 'the ocean triad hypothesis'.

Dynamics of two picophytoplankton groups in mediterranean sea: analysis of the deep chlorophyll maximum by a stochastic advection-reaction-diffusion model.

A stochastic advection-reaction-diffusion model with terms of multiplicative white Gaussian noise, valid for weakly mixed waters, is studied to obtain the vertical stationary spatial distributions of two groups of picophytoplankton, i.e., picoeukaryotes and Prochlorococcus, which account about for 60% of total chlorophyll on average in Mediterranean Sea. By numerically solving the equations of the model, we analyze the one-dimensional spatio-temporal dynamics of the total picophytoplankton biomass and nutrient concentration along the water column at different depths. In particular, we integrate the equations over a time interval long enough, obtaining the steady spatial distributions for the cell concentrations of the two picophytoplankton groups. The results are converted into chlorophyll a and divinil chlorophyll a concentrations and compared with experimental data collected in two different sites of the Sicily Channel (southern Mediterranean Sea). The comparison shows that real distributions are well reproduced by theoretical profiles. Specifically, position, shape and magnitude of the theoretical deep chlorophyll maximum exhibit a good agreement with the experimental values.