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.

The hepatotoxic fluoroquinolone trovafloxacin disturbs TNF- and LPS-induced p65 nuclear translocation in vivo and in vitro.

Idiosyncratic drug-induced liver injury (IDILI) is a severe disease that cannot be detected during drug development. It has been shown that hepatotoxicity of some compounds associated with IDILI becomes apparent when these are combined in vivo and in vitro with LPS or TNF. Among these compounds trovafloxacin (TVX) induced apoptosis in the liver and increased pro-inflammatory cytokines in mice exposed to LPS/TNF. The hepatocyte survival and the cytokine release after TNF/LPS stimulation relies on a pulsatile activation of NF-κB. We set out to evaluate the dynamic activation of NF-κB in response to TVX + TNF or LPS models, both in mouse and human cells. Remarkably, TVX prolonged the first translocation of NF-κB induced by TNF both in vivo and in vitro. The prolonged p65 translocation caused by TVX was associated with an increased phosphorylation of IKK and MAPKs and accumulation of inhibitors of NF-κB such as IκBα and A20 in HepG2. Coherently, TVX suppressed further TNF-induced NF-κB translocations in HepG2 leading to decreased transcription of ICAM-1 and inhibitors of apoptosis. TVX prolonged LPS-induced NF-κB translocation in RAW264.7 macrophages increasing the secretion of TNF. In summary, this study presents new, relevant insights into the mechanism of TVX-induced liver injury underlining the resemblance between mouse and human models. In this study we convincingly show that regularly used toxicity models provide a coherent view of relevant pathways for IDILI. We propose that assessment of the kinetics of activation of NF-κB and MAPKs is an appropriate tool for the identification of hepatotoxic compounds during drug development.

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.

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.

[Inequality in the prevalence of intestinal parasitic infections among schoolchildren from urban and rural schools]. / Desigualdad en la prevalencia de parasitosis intestinal en escolares de una escuela urbana y dos rurales de la comuna de Puerto Montt.

BACKGROUND: Parasitic infections in children reflect social inequalities throughout the world, especially in urban and rural territories. AIM: To assess inequalities in the prevalence of infections by protozoa and intestinal helminths and associated factors in children of different geographical areas from the commune of Puerto Montt. MATERIAL AND METHODS: Cross-sectional study carried out in 103 students from one urban and two rural schools in the commune of Puerto Montt. A socio-demographic survey was applied and the presence parasitic infections was evaluated in serial stool samples. RESULTS: The prevalence of Protozoan and intestinal helminthic infections were 37.5 and 68.1% in urban and rural areas respectively. A regression analysis showed that the differences between rural and urban children, disappear when the results are adjusted by family income and the quality of sewage disposal. In urban children, the prevalence of parasitic infections increases along with the decrease in family income. CONCLUSIONS: Income and sanitary conditions rather than being or urban or rural origin explain the variations in rates of childhood parasitic infections. Infections with protozoa and intestinal helminths are an indicator of social inequality in health.

Desigualdad en la prevalencia de parasitosis intestinal en escolares de una escuela urbana y dos rurales de la comuna de Puerto Montt / Inequality in the prevalence of intestinal parasitic infections among schoolchildren from urban and rural schools

Background: Parasitic infections in children reflect social inequalities throughout the world, especially in urban and rural territories. Aim: To assess inequalities in the prevalence of infections by protozoa and intestinal helminths and associated factors in children of different geographical areas from the commune of Puerto Montt. Material and Methods: Cross-sectional study carried out in 103 students from one urban and two rural schools in the commune of Puerto Montt. A socio-demographic survey was applied and the presence parasitic infections was evaluated in serial stool samples. Results: The prevalence of Protozoan and intestinal helminthic infections were 37.5 and 68.1% in urban and rural areas respectively. A regression analysis showed that the differences between rural and urban children, disappear when the results are adjusted by family income and the quality of sewage disposal. In urban children, the prevalence of parasitic infections increases along with the decrease in family income. Conclusions: Income and sanitary conditions rather than being or urban or rural origin explain the variations in rates of childhood parasitic infections. Infections with protozoa and intestinal helminths are an indicator of social inequality in health.

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.

Assessment of mercury exposure in human populations: A status report from Augusta Bay (southern Italy).

Here we investigate mercury concentrations in the blood (HgB), urine (HgU) and human hair (HgH) of 224 individuals from a coastal area (Eastern Sicily, SE Italy) strongly affected by Hg contamination from one of the largest chlor-alkali plants in Europe. The factors affecting the distribution of Hg and the extent of the exposure of individuals have been explored with a multidisciplinary approach. Multiple regression analyses, together with evidence of high levels of HgB (exceeding the HBMI recommended levels in 50% of cases) and HgH (exceeding the EPA reference dose in 70% of cases), primarily suggest that the consumption of local fish is the main source of Hg for humans. no. significant exposure to inorganic mercury was identified. Toxicokinetic calculations produced a provisional tolerable weekly intake (PTWI) level that, in most cases, exceeds international recommendations, particularly for residents in the studied area.

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'.

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.

The key role played by the Augusta basin (southern Italy) in the mercury contamination of the Mediterranean Sea.

The Augusta basin, located in SE Sicily (southern Italy), is a semi-enclosed marine area, labelled as a highly contaminated site. The release of mercury into the harbour seawater and its dispersion to the blue water, make the Augusta basin a potential source of anthropogenic pollution for the Mediterranean Sea. A mass balance was implemented to calculate the HgT budget in the Augusta basin. Results suggest that an average of ∼0.073 kmol of HgT is released, by diffusion, on a yearly basis, from sediments to the seawater, with a consequent output of 0.162 kmol y(-1) to coastal and offshore waters; this makes the Augusta area an important contributor of mercury to the Mediterranean Sea. Owing to the geographical location of the Augusta basin, its outflowing shelf-waters are immediately intercepted by the surface Atlantic Ionian Stream (AIS) and mixed with the main gyres of the eastern Mediterranean Sea, thus representing a risk for the large-scale marine system.