Effects of marine noise pollution on Mediterranean fishes and invertebrates: A review.

Marine noise pollution (MNP) can cause a multitude of impacts on many organisms, but information is often scattered and general outcomes difficult to assess. We have reviewed the literature on MNP impacts on Mediterranean fish and invertebrates. Both chronic and acute MNP produced by various human activities - e.g. maritime traffic, pile driving, air guns - were found to cause detectable effects on intra-specific communication, vital processes, physiology, behavioral patterns, health status and survival. These effects on individuals can extend to inducing population- and ecosystem-wide alterations, especially when MNP impacts functionally important species, such as keystone predators and habitat forming species. Curbing the threats of MNP in the Mediterranean Sea is a challenging task, but a variety of measures could be adopted to mitigate MNP impacts. Successful measures will require more accurate information on impacts and that effective management of MNP really becomes a priority in the policy makers' agenda.

MALDI-MS and NanoSIMS imaging techniques to study cnidarian-dinoflagellate symbioses.

Cnidarian-dinoflagellate photosynthetic symbioses are fundamental to biologically diverse and productive coral reef ecosystems. The hallmark of this symbiotic relationship is the ability of dinoflagellate symbionts to supply their cnidarian host with a wide range of nutrients. Many aspects of this association nevertheless remain poorly characterized, including the exact identity of the transferred metabolic compounds, the mechanisms that control their exchange across the host-symbiont interface, and the precise subcellular fate of the translocated materials in cnidarian tissues. This lack of knowledge is mainly attributed to difficulties in investigating such metabolic interactions both in situ, i.e. on intact symbiotic associations, and at high spatial resolution. To address these issues, we illustrate the application of two in situ and high spatial resolution molecular and ion imaging techniques-matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI-MSI) and the nano-scale secondary-ion mass spectrometry (NanoSIMS) ion microprobe. These imaging techniques provide important new opportunities for the detailed investigation of many aspects of cnidarian-dinoflagellate associations, including the dynamics of cellular interactions.

The transcriptomic response to thermal stress is immediate, transient and potentiated by ultraviolet radiation in the sea anemone Anemonia viridis.

Among the environmental threats to coral reef health, temperature and ultraviolet increases have been proposed as major agents, although the relative contribution of each in the cnidarian/zooxanthellae symbiosis breakdown has been poorly addressed. We have investigated the transcriptomic response to thermal stress, with and without ultraviolet radiation (UVR), in the symbiotic sea anemone Anemonia viridis. Using the Oligo2K A. viridis microarray, dedicated to genes potentially involved in the symbiosis interaction, we monitored the gene expression profiles after 1, 2 and 5 days of stresses that further lead to massive losses of zooxanthellae. Each stress showed a specific gene expression profile with very little overlap. We showed that the major response to thermal stress is immediate (24 h) but returns to the baseline gene expression profile after 2 days. UVR alone has little effect but potentiates thermal stress, as a second response at 5 days was observed when the two stresses were coupled. Several pathways were highlighted, such as mesoglea loosening, cell death and calcium homeostasis and described in more details. Finally, we showed that the dermatopontin gene family, potentially involved in collagen fibrillogenesis, issued from actinarian-specific duplication events, with one member preferentially expressed in the gastroderm and specifically responding to stress. Anemonia viridis EST sequences have been deposited into GenBank dbEST ([GenBank:FK719875­FK759813].

Microarray analysis of cytochrome P450 mediated insecticide resistance in Drosophila.

Insecticide resistance in laboratory selected Drosophila strains has been associated with upregulation of a range of different cytochrome P450s, however in recent field isolates of D. melanogaster resistance to DDT and other compounds is conferred by one P450 gene, Cyp6g1. Using microarray analysis of all Drosophila P450 genes, here we show that different P450 genes such as Cyp12d1 and Cyp6a8 can also be selected using DDT in the laboratory. We also show, however, that a homolog of Cyp6g1 is over-expressed in a field resistant strain of D. simulans. In order to determine why Cyp6g1 is so widely selected in the field we examine the pattern of cross-resistance of both resistant strains and transgenic flies over-expressing Cyp6g1 alone. We show that all three DDT selected P450s can confer resistance to the neonicotinoid imidacloprid but that Cyp6a8 confers no cross-resistance to malathion. Transgenic flies over-expressing Cyp6g1 also show cross-resistance to other neonicotinoids such as acetamiprid and nitenpyram. We suggest that the broad level of cross-resistance shown by Cyp6g1 may have facilitated its selection as a resistance gene in natural Drosophila populations.

Overproduction of a P450 that metabolizes diazinon is linked to a loss-of-function in the chromosome 2 ali-esterase (MdalphaE7) gene in resistant house flies.

Up-regulation of detoxifying enzymes in insecticide-resistant strains of the house fly is a common mechanism for metabolic resistance. However, the molecular basis of this increased insecticide metabolism is not well understood. In the multiresistant Rutgers strain, several cytochromes P450 and glutathione S-transferases are constitutively overexpressed at the transcriptional level. Overexpression is the result of trans-regulation, and a regulatory gene has been located on chromosome 2. A Gly137 to Asp point mutation in alphaE7 esterase gene, leading to the loss of carboxylesterase activity, has been associated with organophosphate resistance in the house fly and the sheep blowfly. We show here that purified recombinant CYP6A1 is able to detoxify diazinon with a high efficiency. We also show that either the Gly137 to Asp point mutation in alphaE7 esterase gene or a deletion at this locus confer resistance and overproduction of the CYP6A1 protein. Based on these findings, we propose it is the absence of the wild-type Gly137 allele of the alphaE7 gene that releases the transcriptional repression of genes coding for detoxification enzymes such as CYP6A1, thereby leading to metabolic resistance to diazinon.

Tissue-specific induction and inactivation of cytochrome P450 catalysing lauric acid hydroxylation in the sea bass, Dicentrarchus labrax.

Microsomal cytochrome P450-dependent lauric acid hydroxylase activities were characterized in liver, kidney, and intestinal mucosa of the sea bass (Dicentrarchus labrax). Microsomes from these organs generated (omega-1)-hydroxylauric acid and a mixture of positional isomers including (omega)-, (omega-2)-, (omega-3)- and (omega-4)-hydroxylauric acids, which were identified by RP-HPLC and GC-MS analysis. Peroxisome proliferators, such as clofibrate and especially di(2-ethylhexyl) phthalate, increased kidney microsomal lauric acid hydroxylase activities. The synthesis of 11-hydroxylauric acid was enhanced 5.3-fold in kidney microsomes. Liver microsomal lauric acid hydroxylase activities were weakly affected and no significant induction was found in small intestine microsomes from clofibrate or di(2-ethylhexyl) phthalate-treated fish. The differences in lauric acid metabolisation and the tissue-specific induction by peroxisome proliferators suggest the involvement of several P450s in this reaction. Incubations of liver and kidney microsomes with lauric acid analogues (11- or 10-dodecynoic acids) resulted in a time- and concentration-dependent loss of lauric acid hydroxylase activities. The induction of these activities in fish by phthalates, which are widely-distributed environmental pollutants, may be taken into consideration for the development of new biomarkers.

Cloning of CYP4F7, a kidney-specific P450 in the sea bass Dicentrarchus labrax.

A cDNA sequence coding for a cytochrome P450 of the CYP4F subfamily was isolated from total RNA of sea bass kidney by rapid amplification of cDNA ends. The full length sequence coded for a protein of 526 amino acids. The amino acid sequence shared 39% to 56% residue identities with the mammalian CYP4F sequences, and thus was named CYP4F7 (accession number AF123541). RNA blot analysis using CYP4F7 cDNA as a probe indicated that the corresponding mRNA was only detected in kidney. Expression in the kidney was constitutive, and no induction of this mRNA was detected in this or other tissues, with any of the inducers tested, including peroxisome proliferators.

Molecular cloning of a phthalate-inducible CYP4 gene (CYP4T2) in kidney from the sea bass, Dicentrarchus labrax.

A full-length cDNA sequence was isolated from kidney total RNA of di(2-ethylhexyl) phthalate-treated sea bass by reverse-transcriptase polymerase chain reaction and then rapid amplification of cDNA ends. The deduced amino acid sequence, which has been named CYP4T2, shared 69 and 54.4% amino acid identity with rainbow trout CYP4T1 and rat CYP4B1, respectively. RNA blot analysis using the CYP4T2 cDNA as a probe indicated that the mRNA was rather abundant in kidney, and less so in liver, small intestine, and brain. Treatment of sea bass with peroxisome proliferators showed marked tissue-specific induction. CYP4 inducers clofibrate, di(2-ethylhexyl) phthalate (DEHP), and 2,4-dichlorophenoxy acetic acid (2,4-D) were administered by intraperitoneal injection. The strongest induction was found in kidney after a DEHP treatment (6.5-fold) or a 2,4-D treatment (9-fold), while no induction was observed in liver.