Effects of cadmium, zinc and nitrogen status on non-protein thiols in the macroalgae Enteromorpha spp. from the Scheldt Estuary (SW Netherlands, Belgium) and Thermaikos Gulf (N Aegean Sea, Greece)

Enteromorpha prolifera (Scheldt Estuary) and E. linza (Thermaikos Gulf) were incubated at three salinities with 100 and 200microgL(-1)Cd and Zn. The objective was to measure effects of Cd, Zn and nitrogen (N) status on the pools of metal-binding non-protein thiols: glutathione and phytochelatins, (gamma-glutamyl-cysteinyl)(n)-glycine (PC). In E. linza, ammonium pools were higher, but amino acid pools, total N and protein contents were lower than in E. prolifera. Reduced glutathione (GSH) pools were positively correlated with free glutamate and protein contents. In E. linza GSH pools increased and the ratio of reduced to oxidized glutathione (GSH:(GSH+0.5GSSG)), an indicator of oxidative stress, decreased with Cd contents, indicating Cd-induced glutathione oxidation. Total glutathione pools (reduced plus oxidized) ranged from 16nmolSgdwt(-1) in controls (at 0.5micromolCdgdwt(-1)) to 179nmolSgdwt(-1) (at 1.9micromolCdgdwt(-1)) at the highest cadmium dosage. Cadmium stimulated PC synthesis in E. prolifera which suggests that in N-rich algae, glutathione pools were high enough for PC synthesis. In both species GSH and protein increased with Zn contents, whereas GSH:(GSH+0.5GSSG) decreased, which would indicate Zn-induced oxidative stress; in E. linza, at the highest salinity the glutathione redox ratio decreased from 0.61 (at 2.9micromolZngdwt(-1)) to 0.26 (at 4.9nmolSgdwt(-1)) (at 0.5molCdgdwt(-1)). PCs were not synthesized in response to Zn, which may have resulted in Zn-induced GSH oxidation. The presence of both oxidative effects (Cd, Zn) and detoxification (Cd) could be identified by observing the responses of glutathione and PC pools to metal stress.     

Lead contamination of small cetaceans in European waters--the use of stable isotopes for identifying the sources of lead exposure

Lead concentrations and isotopic composition have been measured in bone and teeth of small cetaceans belonging to three species (Delphinus delphis, Phocoena phocoena and Stenella coeruleoalba), to evaluate the toxicological risk and to determine sources of lead in the European waters. Lead concentrations, far lower than threshold value inducing toxic effects in human, were higher in teeth than in bones, but highly correlated between the two tissues (r=0.92, p<0.001). Large variations of 206Pb/207Pb values in bone tissue showed that cetaceans must be submitted to various atmospheric influences. No geographical differences appeared which is consistent with studies on their distribution indicating seasonal movements between Brittany waters and the Bay of Biscay. The negative correlation between 206Pb/207Pb ratios and age of the individuals reflected the decrease in the production of alkyl lead in Europe, i.e., the increasing use of unleaded gasoline.     

The amphipod Talitrus saltator as a bioindicator of human trampling on sandy beaches

The present paper assesses the use of the supralittoral amphipod Talitrus saltator as a bioindicator of the effects of human trampling on the supralittoral sandy band. Samplings in delimited areas were carried out at sites subjected to different human impact. The results showed a strong negative correlation between the number of swimmers and the sandhopper population density, while there was no clear relationship between sandhopper abundance and the other factors considered: granulometry, compactness and organic carbon content of the sand, and trace metal contents in the sand and sandhoppers. A field test of trampling conducted in a confined space showed its direct negative effect on sandhopper survival. However, trace metal analysis confirmed the ability of T. saltator to bioaccumulate some elements (Hg, Zn, Cu, Cd). Our study demonstrates that T. saltator is a good bioindicator of human impact in the supralittoral zone of sandy shores.     

Domoic acid excretion in dungeness crabs, razor clams and mussels

Domoic acid (DA) is a neurotoxic amino acid produced by several marine algal species of the Pseudo-nitzschia (PN) genus. We studied the elimination of DA from hemolymph after intravascular (IV) injection in razor clams (Siliqua patula), mussels (Mytilus edulis) and Dungeness crabs (Cancer magister). Crabs were also injected with two other organic acids, dichloroacetic acid (DCAA) and kainic acid (KA). For IV dosing, hemolymph was repetitively sampled and DA concentrations measured by HPLC-UV. Toxicokinetic analysis of DA in crabs suggested most of the injected dose remained within hemolymph compartment with little extravascular distribution. This observation is in sharp contrast to results obtained from clams and mussels which exhibited similarly large apparent volumes of distribution despite large differences in overall clearance. These findings suggest fundamentally different storage and elimination processes are occurring for DA between bivalves and crabs.     

Use of immunofluorescence technique in cultured fibroblasts from Mediterranean cetaceans as new "in vitro" tool to investigate effects of environmental contaminants

The aim of the present study was to propose the immunofluorescence technique in cultured fibroblasts from Mediterranean cetaceans as a new "in vitro" tool to explore the susceptibility of these marine mammals to different xenobiotic compounds. The cell lines were cultured from integument biopsies of free-ranging and stranded cetaceans (dead within 12h). Using the indirect immunofluorescence assay, we detected endogenous proteins induced by different contaminants. Here we present the method used for qualitative and quantitative evaluation of cytochromes P450 (CYP1A1 and CYP2B) induced by some POPs (DDTs and PCBs) and emerging contaminants (PBDEs) in fibroblast cell cultures of striped dolphin (Stenella coeruleoalba) and bottlenose dolphin (Tursiops truncatus). Immunofluorescence was quantified with a specially designed Olympus macro, DetectIntZ. A major result was the possibility of using this "in vitro" assay to quantify induction of endogenous proteins.     

The role of water mass dynamics in controlling bacterial abundance and production in the middle Adriatic Sea

Month-to-month fluctuations in the abundance of bacteria and heterotrophic nanoflagellates (HNF) and bacterial production, as well as various chemical (nutrients, oxygen) and physical (salinity, temperature) parameters were analysed at a station located in the open middle Adriatic Sea during one decade (1997-2006). Being influenced by both coastal waters and open Adriatic circulation in the surface layer, and by the deep Adriatic water masses in the deep layers (100 m), this station is quite suitable for detecting the environmental changes occurring in the open Adriatic Sea with respect to the circulation of its water masses and their long-term changes and anomalies. Multivariate methods were used to identify seasonal and inter-annual changes of the investigated parameters, associating observed changes to the changes in Adriatic water masses and circulation regimes. The analyses showed that bacterial abundance and production were controlled by different water mass dynamics during 1997-2001 compared to 2002-2006 period, particularly noticeable in different seasonal patterns of biological parameters. The interplay between North Adriatic Dense Water (NAdDW) and Levantine Intermediate Water (LIW) resulted in a change in the available nutrients (NAdDW is poor in orthophosphates), and as a consequence different bacterial abundance and production. A few periods were examined in detail, such as 2004, when LIW inflow was particularly strong and was accompanied by an increase of bacterial and HNF abundances, as well as of bacterial production.     

Diagnosis of trace metal contamination in sediments: the example of Ensenada and El Sauzal, two harbors in Baja California, Mexico

Total metal concentrations in sediments from within Ensenada and El Sauzal Harbors are generally higher than at the mouths. Grain-size analyses suggested that this enrichment could be due to the presence of fine-grained sediments in the inner part of the harbors rather than to anthropogenic perturbations. The (Me/Al)(sample) ratios for Pb, Co, Ni and Fe were significantly higher for Ensenada Harbor relative to El Sauzal Harbor, whereas the ratios for Cd, Mn, Zn and Cu were statistically equivalent for both harbors. Calculated enrichment factors [EF(Me)=(Me/Al)(sample)/(Me/Al)(shale)] indicated that the metals showing slight enrichment were those associated with anthropogenic contamination (Pb, Zn), or probably related to primary productivity in the water column (Cd, Co). The levels of most of the metals were not greatly enriched, a consideration that is of the utmost importance when contamination issues are at stake.     

The influence of 70 and 120 kHz tonal signals on the behavior of harbor porpoises (Phocoena phocoena) in a floating pen

Two harbor porpoises in a floating pen were subjected to five pure tone underwater signals of 70 or 120kHz with different signal durations, amplitudes and duty cycles (% of time sound is produced). Some signals were continuous, others were intermittent (duty cycles varied between 8% and 100%). The effect of each signal was judged by comparing the animals' surfacing locations and number of surfacings (i.e. number of respirations) during test periods with those during baseline periods. In all cases, both porpoises moved away from the sound source, but the effect of the signals on respiration rates was negligible. Pulsed 70kHz signals with a source level (SL) of 137dB had a similar effect as a continuous 70kHz signal with an SL of 148dB (re 1muPa, rms). Also, a pulsed 70kHz signal with an SL of 147dB had a much stronger deterring effect than a continuous 70kHz signal with a similar SL. For pulsed 70kHz signals (2s pulse duration, 4s pulse interval, SL 147dB re 1muPa, rms), the avoidance threshold sound pressure level (SPL), in the context of the present study, was estimated to be around 130dB (re 1muPa, rms) for porpoise 064 and around 124dB (re 1muPa, rms) for porpoise 047. This study shows that ultrasonic pingers (70kHz) can deter harbor porpoises. Such ultrasonic pingers have the advantage that they do not have a "dinner bell" effect on pinnipeds, and probably have no, or less, effect on other marine fauna, which are often sensitive to low frequency sounds.     

Anoxia over the western continental shelf of India: bacterial indications of intrinsic nitrification feeding denitrification

Studies on the Arabian Sea coastal anoxia have been of immense interest, but despite its ecological significance there is sparse understanding of the microbes involved. Hence, observations were carried out off Goa (15 degrees 30'N, 72 degrees 40'E to 15 degrees 30'N, 72 degrees 59'E) to understand the processes that mediate the changes in various inorganic nitrogen species in the water column during anoxia. Water column chemistry showed a clear distinct oxic environment in the month of April and anoxic condition in October. Our study based on microbial signatures indicated that oxygen deficit appeared as a well-defined nucleus almost 40 km away from the coast during the oxic period (April) and spreads there after to the entire water column synchronizing with the water chemistry. Striking results of net changes in inorganic nitrogen species in nitrification blocked and unblocked experimental systems show that denitrification is the predominant process in the water column consuming available nitrate ( approximately 0.5 microM) to near zero levels within approximately 72 h of incubation. These observations have been supported by concomitant increase in nitrite concentration ( approximately 4 microM). Similar studies on denitrification-blocked incubations, demonstrate the potential of nitrification to feed denitrification. Nitrification could contribute almost 4.5 microM to the total nitrate pool. It was found that the relation between ammonium and total dissolved inorganic nitrogen (DIN) pool (r=0.98, p<0.001, n=122) was significant compared to the latter with nitrite and nitrate. The occurrence of high ammonium under low phosphate conditions corroborates our observations that ammonium does not appear to be locked under low oxygen regimes. It is suggested that ammonium actively produced by detrital breakdown (ammonification) is efficiently consumed through nitrification process. The three processes in concert viz. ammonification, nitrification and denitrification appear to operate in more temporal and spatial proximity than hitherto appreciated in these systems and this gives additional cues on the absence of measurable nitrate at surface waters, which was earlier attributed only to efficient algal uptake. Hence we hypothesize that the alarming nitrous oxide input into the atmosphere could be due to high productivity driven tighter nitrification-denitrification coupling, rather than denitrification driven by extraneous nitrate.     

Habitat continuity effects on gradients of fish biomass across marine protected area boundaries

Marine protected areas (MPAs) could be useful as fisheries management tools for the exportation of pelagic eggs, larvae and adult fish. A decreasing gradient of fish biomass across MPAs boundary may indicate export. We determine whether gradients of decreasing biomass of fish assemblage occurred in Tabarca Marine Reserve over two habitats with different continuity across the boundaries, to test if the patchy nature of the marine environment might act as a barrier for the fish export. In general, significant decreasing gradients in total fish biomass and biomass of some species were observed on P. oceanica and rocky substrates, independently of their different continuity through the reserve boundaries. Changes in the multivariate structure of the fish assemblage were correlated with the distance from integral reserve. All of these results support the hypothesis that the exportation of adult fish from Tabarca Marine Reserve occurs, and this process may influence the surrounding fished areas.