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Recent studies have suggested that Mediterranean indigenous fish species are affected by bioactive metabolites coming from marine invasive species via food web interactions. In particular, both physiological and behavioural changes in the white sea bream Diplodus sargus were related to caulerpin (CAU), a bisindolic alkaloid particularly abundant in the invasive alga Caulerpa cylindracea, on which the fish actively feed. Dietary administration of CAU decreased aggressiveness in D. sargus, suggesting an anxiolytic-like effect of CAU possibly mediated by endogenous anxiolytic agents. This hypothesis is supported here by the finding of a significant increase of NPY transcriptional expression in the brain of fish fed with CAU enriched food, shedding more light on the neural mechanisms behind the altered behaviour of D. sargus. Fundação para a Ciência e a Tecnologia - FCT; FEDER info:eu-repo/semantics/publishedVersion

The biological invasion of the green algae Caulerpa cylindracea represents a serious scientific and public issue in the Mediterranean Sea, essentially due to strong modifications both to habitat structure and native benthic communities. Although alterations in health status and changes in flesh quality of some marine species (dietary exposed to C. cylindracea) have been observed, no studies on cause-effect relationships have been carried out. Here, for the first time, through a controlled feeding experiment followed by 1H NMR Spectroscopy and multivariate analysis (PCA, OPLS-DA), we showed that caulerpin taken with diet is directly responsible of changes observed in metabolic profile of fish flesh, including alteration of lipid metabolism, in particular with a reduction of &omega, 3 PUFA content. The potential of caulerpin to directly modulate lipid metabolism opens up new questions about causal mechanism triggered by algal metabolite also in view of a possible exploitation in the nutraceutical/medical field.

The green alga Caulerpa cylindracea is a non-autochthonous and invasive species that is severely affecting the native communities in the Mediterranean Sea. Recent researches show that the native edible fish Diplodus sargus actively feeds on this alga and cellular and physiological alterations have been related to the novel alimentary habits. The complex effects of such a trophic exposure to the invasive pest are still poorly understood. Here we report on the metabolic profiles of plasma from D. sargus individuals exposed to C. cylindracea along the southern Italian coast, using 1H NMR spectroscopy and multivariate analysis (Principal Component Analysis, PCA, Orthogonal Partial Least Square, PLS, and Orthogonal Partial Least Square Discriminant Analysis, OPLS-DA). Fish were sampled in two seasonal periods from three different locations, each characterized by a different degree of algal abundance. The levels of the algal bisindole alkaloid caulerpin, which is accumulated in the fish tissues, was used as an indicator of the trophic exposure to the seaweed and related to the plasma metabolic profiles. The profiles appeared clearly influenced by the sampling period beside the content of caulerpin, while the analyses also supported a moderate alteration of lipid and choline metabolism related to the Caulerpa-based diet.

Biological invasions are deemed as one of the most severe threats to biodiversity and can result in huge economic impacts on natural resources. Many studies have assessed structural changes caused by the introduction of nonindigenous species, but urgent questions remain to be answered about the complex indirect effects of invasive pests on marine biodiversity at the species and ecosystem level. Previous investigations focusing on the biological invasion by Caulerpa racemosa in the Mediterranean indicated that the lipophilic algal metabolite caulerpin accumulates in the tissues of the native edible white sea bream Diplodus sargus feeding on the exotic alga. The level of caulerpin in D. sargus has been thus used as an indicator of the trophic exposure of the fish to the seaweed and related with adverse effects on the fish health. In the present study, we show that the C. racemosa-based diet can also alter muscle fatty acids composition in the white sea bream by reducing the percentage of polyunsaturated fatty acids of the n-3 and n-6 series, such as eicosapentaenoic (C20:5), docosahexaenoic (C22:6) and arachidonic acids (C20:4). This represents both a real threat to the health of fish unable to biosynthesize these essential fatty acids and an impoverishment of an important source of essential fatty acids for human nutrition, which helps to prevent cardiovascular diseases, diabetes, hepatic steatosis, cancer and neurological disorders. On the whole, this work sheds light on an unexplored and critical aspect of biological invasions with implication on the health of both humans and the environment. © 2014 Springer-Verlag Berlin Heidelberg.

The green alga Caulerpa racemosa is a non-native, invasive species in the Mediterranean, and an important stressor for several native organisms. The algal capacity to produce secondary metabolites has been suggested to modulate success of the C. racemosa invasion, although many of potentially involved biological pathways still remain unexplored. In this respect, the aim of the present study was to investigate some molecular and cellular effects in the white seabream Diplodus sargus , an ecologically key species, which included the alien C. racemosa in its diet. Organisms were sampled in 2 seasonal periods from 3 locations of Southern Italy, each characterized by different levels of algal abundance. The level of caulerpin, one of the main secondary algal metabolites, in fish tissues has been used as an indicator of the trophic exposure to the seaweed and related with molecular and cellular responses. Chemical analyses indicated that fish from invaded sites can accumulate caulerpin, with liver concentrations ranging from a few up to hundreds of μg/g. Biomarkers analyses revealed only limited alterations of the main antioxidant defences, such as glutathione reductase and levels of glutathione; on the other hand, increased enzymatic activities of cytochrome P450, glutathione S-transferases and acyl CoA oxidase, as well as enhanced gene transcription for peroxisome proliferator-activated receptor alpha, cytochrome P4501A and vitellogenin 1 were observed in fish more exposed to C. racemosa as indicated by liver concentrations of caulerpin higher than 50 μg/g. Despite a direct molecular relationship with this algal metabolite could not be established, our results suggest that a C. racemosa enriched diet can modulate biotransformation and fatty acids metabolism of D. sargus. Assessing whether similar effects represent short- or long-term effects will be of crucial importance to understand consequences on the general health status and reproductive performance of exposed key fish species in the Mediterranean region.

The green alga Caulerpa racemosa var. cylindracea has invaded Mediterranean seabed including marine reserves, modifying the structure of habitats and altering the distributional patterns of associated organisms. However, the understanding of how such invasion can potentially affect functional properties of Mediterranean subtidal systems is yet to be determined. In this study, we show that C. racemosa changes foraging habit of the native white seabream, Diplodus sargus. In invaded areas, we found a high frequency of occurrence of C. racemosa in the stomach contents of this omnivorous fish (72.7 and 85.7%), while the alga was not detected in fish from a control area. We also found a significant accumulation of caulerpin, one of the main secondary metabolites of C. racemosa, in fish tissues. The level of caulerpin in fish tissues was used here as an indicator of the trophic exposure to the invasive pest and related with observed cellular and physiological alterations. Such effects included activation of some enzymatic pathways (catalase, glutathione peroxidases, glutathione S-transferases, total glutathione and the total oxyradical scavenging capacity, 7-ethoxy resorufin O-deethylase), the inhibition of others (acetylcholinesterase and acylCoA oxidase), an increase of hepatosomatic index and decrease of gonadosomatic index. The observed alterations might lead to a detrimental health status and altered behaviours, potentially preventing the reproductive success of fish populations. Results of this study revealed that the entering of alien species in subtidal systems can alter trophic webs and can represent an important, indirect mechanism which might contribute to influence fluctuations of fish stocks and, also, the effectiveness of protection regimes.

S100β protein and glial fibrillary acidic protein (GFAP) are widely distributed in astrocytes1 but also in ependymal and radial glia1. These proteins are then used as reliable markers of all astroglial cells. In teleost fishes, the studies on the astroglia are still relatively scarce. In this paper we report the study on the astroglial architecture in the brain of a teleost fish, Diplodus sargus, by the immunohistochemical staining of S100β protein and GFAP. The immunodetection of GFAP and S100 showed the presence of an astroglial architecture various in the sargus brain. It was represented mainly by radial glia and fibres, which were more numerous and large in the mesencephalon and rhombencephalon, but very scarce in the telencephalon. In fact in this region we revealed few and thin fibres immunoreactive only to anti-GFAP. In the other regions of the brain indeed we observed structures both GFAP(+) and S100(+). The immunodetection of these protein revealed generally a major number of GFAP(+) structures occasionally also S100(+). However, the immunolabeling of GFAP was more strong than the S100 immunolabeling. Radial glia was especially evident in the optic tectum and in the white matter of the cerebellum. The occurrence of bundles of GFAP(+) fibres, but also S100(+), was observed in the periventricular region, in the inferior lobes of the hypothalamus, in the dorsal thalamus and in the optic tectum. The infudibular region and the neurohypophysis showed also intensely GFAP(+) fibres, processes of the pituicytic cells, which appeared also S100(+).

In the last decades, marine reserves have dramatically increased in number worldwide. Here I examined the potential of no-take marine reserves to reestablish lost predatory interactions and, in turn, cause community-wide changes in Mediterranean rocky reefs. Protected locations supported higher density and size of the most effective fish preying on sea urchins (the sea breams Diplodussargus and D. vulgaris) than unprotected locations. Density of sea urchins (Paracentrotus lividus and Arbacia lixula) was lower at protected than at unprotected locations. Size structure of P. lividus was bimodal (a symptom of predation on medium-sized urchins) only at the protected locations. Coralline barrens were less extended at protected than at unprotected locations, whereas turf-forming and erect-branched algae showed an opposite pattern. Erect-unbranched and erect-calcified algae and conspicuous zoobenthic organisms did not show any pattern related to protection. Tethering experiments showed that predation impact on urchins was (1) higher at protected than at unprotected locations, (2) higher on P. lividus than on A. lixula, and (3) higher on medium-sized (2-3.5 cm test diameter) than large-sized (> 3.5 cm) urchins. Sea urchins preyed on by fish in natural conditions were smaller at unprotected than at protected locations. The analysis of sea urchin remains found in Diplodus fish stomachs revealed that medium-sized P. lividus were the most frequently preyed upon urchins and that size range of consumed sea urchins expanded with increasing size of Diplodus fish. These results suggest that (1) depletion and size reduction of predatory fish caused by fishing alter patterns of predation on sea urchins, and that (2) fishing bans (e.g., within no-take marine reserves) may reestablish lost interactions among strongly interactive species in temperate rocky reefs with potential community-wide effects.