The effect of low frequency noise on the behaviour of juvenile Sparus aurata.

Anthropogenic activities are causing increased noise levels in the marine environment. To date, few studies have been undertaken to investigate the effects of different noise frequencies on the behaviour of juvenile fish. In this study, the behavioural changes of juvenile gilthead seabream (Sparus aurata) are evaluated when exposed to white noise filtered in third-octave bands centred at 63, 125, 500, and 1000 Hz (sound pressure level, 140-150 dB re 1 µΡa) for 7 h. The group dispersion, motility, and swimming height of the fish were analysed before and during the acoustic emission. Dispersion of the fish was found to reduce immediately upon application of low frequency sound (63 and 125 Hz) with a return to control condition after 2 h (indicative of habituation), whereas at 1 kHz, dispersion increased after 2 h without any habituation. The motility decreased significantly at 63 Hz throughout the 7 h of sound exposure. The swimming height decreased significantly for all frequencies other than 125 Hz. The results of this study highlight significant variations in the behavioural responses of juvenile fish that could have consequences on their fitness and survival.

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.

Effects of acoustic stimulation on biochemical parameters in the digestive gland of Mediterranean mussel Mytilus galloprovincialis (Lamarck, 1819).

Underwater sounds generated by anthropogenic activity can cause behavior changes, temporary loss of hearing, damage to parts of the body, or death in a number of marine organisms and can also affect healing and survival. In this study, the authors examined the effects of high-frequency acoustic stimulations on a number of biochemical parameters in the Mediterranean mussel, Mytilus galloprovincialis. During the experiment, animals were placed in a test tank and exposed to acoustic signals [a linear sweep ranging from 100 to 200 kHz and lasting 1 s, with a sound pressure level range of between 145 and 160 dBrms (re 1µParms)] for 3 h. Total haemocyte count was assessed and glucose levels, cytotoxic activity and enzyme activity (alkaline phosphatase, esterase and peroxidase) in the digestive gland were measured. For the first time, this study suggests that high-frequency noise pollution has a negative impact on biochemical parameters in the digestive gland.

Underwater high frequency noise: Biological responses in sea urchin Arbacia lixula (Linnaeus, 1758).

Marine life is extremely sensitive to the effects of environmental noise due to its reliance on underwater sounds for basic life functions, such as searching for food and mating. However, the effects on invertebrate species are not yet fully understood. The aim of this study was to determine the biochemical responses of Arbacia lixula exposed to high-frequency noise. Protein concentration, enzyme activity (esterase, phosphatase and peroxidase) and cytotoxicity in coelomic fluid were compared in individuals exposed for three hours to consecutive linear sweeps of 100 to 200 kHz lasting 1 s, and control specimens. Sound pressure levels ranged between 145 and 160 dB re 1µPa. Coelomic fluid was extracted and the gene and protein expression of HSP70 with RT-PCR was evaluated on coelomocytes. A significant change was found in enzyme activity and in the expression of the HSP70 gene and protein compared to the control. These results suggested that high-frequency stimuli elicit a noise-induced physiological stress response in A. lixula, confirming the vulnerability of this species to acoustic exposure. Furthermore, these findings provide the first evidence that cell-free coelomic fluid can be used as a signal to evaluate noise exposure in marine invertebrates.

Sound production mechanism in the semiterrestrial crab Neohelice granulata (Brachyura, Varunidae).

Very few studies of sound production in the Brachyura have simultaneously identified the type of individuals (e.g., sex) producing acoustic signals, the structures involved in making sound and the social context. The emission and type of sound signals in Neohelice granulata were previously characterized, but the sex and the body structures involved in the sound production mechanism were not determined. In the present study, experiments conducted in the laboratory demonstrated that acoustic signals were produced by males through an up-down movement of the cheliped by rubbing the merus against the pterygostomial area of the carapace. The micromorphology of the merus showed that it has a ridge of tubercles which may act as a plectrum, while the pterygostomial area bears tubercles and might function as the pars stridens. Acoustic signals were displayed more frequently in the presence of receptive females. Agonistic encounters among males also occurred more often in the presence of receptive females. The authors propose that Neohelice granulata males use their chelipeds to produce sound signals in a mating context, probably to attract the receptive female and/or to repel other males when a receptive female is present. Thus, the display might have a reproductive function influencing mate choice.

Acoustic impact of a wave energy converter in Mediterranean shallow waters.

In this study, underwater noise from a full-scale wave energy converter system (ISWEC), installed on the coast of Pantelleria Island (central Mediterranean Sea), was characterized. The noise was measured using an autonomous acoustic recorder anchored to the sea bottom 40 m from the ISWEC hull. Acoustic monitoring continued for 15 months, starting 7 months before (PRE), 2 months during (INST) and 6 months after the ISWEC installation (POST). The levels of noise, assessed with power spectrum density and octave and third-octave band sound pressure levels (BSPLs), were higher during the POST period than during the PRE period at lower frequencies up to 4 kHz and increased with wave height. During the ISWEC activation for energy production (POST_ON) in the wave height range 1-2.9 m, the BSPLs increased much more at lower frequencies up to 4 kHz (the median BSPLs at 63 Hz for the PRE, POST, and POST_ON conditions were 73, 106, and 126 dB re 1µPa, respectively). Considering the biophonies that make up the soundscape of the area, we examined the possible masking of fish choruses due to ISWEC noise and highlighted that at a distance of 1000 m, the 800 Hz peak frequency was 10 dB above the ISWEC signal. Within this distance from ISWEC, a possible masking effect is supposed.

Underwater acoustic communication during the mating behaviour of the semi-terrestrial crab Neohelice granulata.

In semi-terrestrial crabs, the production of sounds has been recognized to be related to courtship communication dynamics. The present study aimed to assess if the crab Neohelice granulata (Varunidae) was able to emit acoustic signals and if they played a role in the crab's behaviour. We also assessed the locomotor behaviours to examine these parameters in different mating contexts of crabs. The study was divided into two different experimental conditions: 'solitary experiment' (consisting of three combination layouts with male, unreceptive and receptive females alone) and 'group experiment' (consisting of mixed combinations layouts of males, unreceptive, and receptive females). Synchronized acoustic and video monitoring systems were used to record the acoustic signals and locomotor behaviours of alone and grouped specimens. The greatest values of locomotor behavioural parameters were observed in layouts with receptive females alone and with 2 males plus 1 receptive female, probably related to courtship behaviour. N. granulata produced two distinct signals, a multi-pulse rasp signal (highest numbers were recorded in layouts with male alone and with 2 males plus 1 receptive female) and a single rasp signal. These results may suggest that males use the multi-rasp signal to advertise their presence to other males or to attract receptive females.

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.

Are semi-terrestrial crabs threatened by human noise? Assessment of behavioural and biochemical responses of Neohelice granulata (Brachyura, Varunidae) in tank.

This study examined the effects of human lab-generated noise (sweep tone) on the behaviour and biochemistry of a semi-terrestrial crab (Neohelice granulata). The experiment was carried out in tanks equipped with video- and audio-recording systems on a total of seventy-eight specimens. In total, 42 experimental trials with sweep-tone exposure and control conditions were performed using crabs in single and group layouts. After a habituation period of 30 min, the locomotor and acoustic (sound signals emitted by the crabs) behaviours were monitored for 30 min. During this time, the animals in sweep-tone conditions were exposed to ascending sweeps in a bandwidth range of 2.5-25 kHz. Exposure to sweep-tone noise produced significant changes in the number of signals emitted, locomotor behaviours and plasma parameters, such as haemolymph total haemocyte count and glucose, lactate and total protein concentrations, revealing that human noise could represent a disturbance for this crustacean species.

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.

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.

Evolutionary conserved mechanisms pervade structure and transcriptional modulation of allograft inflammatory factor-1 from sea anemone Anemonia viridis.

Gene family encoding allograft inflammatory factor-1 (AIF-1) is well conserved among organisms; however, there is limited knowledge in lower organisms. In this study, the first AIF-1 homologue from cnidarians was identified and characterised in the sea anemone Anemonia viridis. The full-length cDNA of AvAIF-1 was of 913 bp with a 5' -untranslated region (UTR) of 148 bp, a 3'-UTR of 315 and an open reading frame (ORF) of 450 bp encoding a polypeptide with149 amino acid residues and predicted molecular weight of about 17 kDa. The predicted protein possesses evolutionary conserved EF hand Ca2+ binding motifs, post-transcriptional modification sites and a 3D structure which can be superimposed with human members of AIF-1 family. The AvAIF-1 transcript was constitutively expressed in all tested tissues of unchallenged sea anemone, suggesting that AvAIF-1 could serve as a general protective factor under normal physiological conditions. Moreover, we profiled the transcriptional activation of AvAIF-1 after challenges with different abiotic/biotic stresses showing induction by warming conditions, heavy metals exposure and immune stimulation. Thus, mechanisms associated to inflammation and immune challenges up-regulated AvAIF-1 mRNA levels. Our results suggest its involvement in the inflammatory processes and immune response of A. viridis.

Long-Term Monitoring of Dolphin Biosonar Activity in Deep Pelagic Waters of the Mediterranean Sea.

Dolphins emit short ultrasonic pulses (clicks) to acquire information about the surrounding environment, prey and habitat features. We investigated Delphinidae activity over multiple temporal scales through the detection of their echolocation clicks, using long-term Passive Acoustic Monitoring (PAM). The Istituto Nazionale di Fisica Nucleare operates multidisciplinary seafloor observatories in a deep area of the Central Mediterranean Sea. The Ocean noise Detection Experiment collected data offshore the Gulf of Catania from January 2005 to November 2006, allowing the study of temporal patterns of dolphin activity in this deep pelagic zone for the first time. Nearly 5,500 five-minute recordings acquired over two years were examined using spectrogram analysis and through development and testing of an automatic detection algorithm. Echolocation activity of dolphins was mostly confined to nighttime and crepuscular hours, in contrast with communicative signals (whistles). Seasonal variation, with a peak number of clicks in August, was also evident, but no effect of lunar cycle was observed. Temporal trends in echolocation corresponded to environmental and trophic variability known in the deep pelagic waters of the Ionian Sea. Long-term PAM and the continued development of automatic analysis techniques are essential to advancing the study of pelagic marine mammal distribution and behaviour patterns.

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.

Benthic Foraminifera as bio-indicators of anthropogenic impacts in coastal environments: Acqua dei Corsari area case study (Palermo, Italy).

This study investigates living benthic foraminiferal assemblages as bio-indicators of anthropogenic activities in a coastal area within the Gulf of Palermo (Sicily, Italy), affected by industrial and urban activities, and evaluates the environmental quality through the calibration of a Tolerant Species index (%TSstd). Sediments from 6 stations were sampled along a bathymetric transect from the coast to offshore. Sediment grain size, TOC, major, minor and trace elements and polycyclic aromatic hydrocarbons (PAHs) were compared to benthic foraminiferal assemblages and species at each station. Diversity and density of benthic foraminiferal assemblages were not affected by the presence of pollutants, while tolerant species increased with organic (TOC and PAHs) or chemical (As and Pb) concentrations. Moreover, the calibration of the %TSstd formula to >125µm foraminiferal assemblage, gives a detailed description of environmental quality along the transect, representing a good and sensitive tool to evaluate marine coastal environment.

Partially Purified Extracts of Sea Anemone Anemonia viridis Affect the Growth and Viability of Selected Tumour Cell Lines.

In the last few years, marine species have been investigated for the presence of natural products with anticancer activity. Using reversed phase chromatography, low molecular weight proteins were fractionated from the sea anemone Anemonia viridis. Four different fractions were evaluated for their cytotoxic activity by means of erythrocyte haemolysis test, MTS, and LDH assays. Finally, the antiproliferative activities of three of these fractions were studied on PC3, PLC/PRF/5, and A375 human cancer cell lines. Our analysis revealed that the four fractions showed different protein contents and diverse patterns of activity towards human PBMC and cancer cell lines. Interestingly, fractions III and IV exerted cytotoxic effects on human cells. Conversely, fractions I and II displayed very low toxic effects associated with antiproliferative activities on cancer cell lines.

Temporal patterns in the soundscape of the shallow waters of a Mediterranean marine protected area.

The study of marine soundscapes is an emerging field of research that contributes important information about biological compositions and environmental conditions. The seasonal and circadian soundscape trends of a marine protected area (MPA) in the Mediterranean Sea have been studied for one year using an autonomous acoustic recorder. Frequencies less than 1 kHz are dominated by noise generated by waves and are louder during the winter; conversely, higher frequencies (4-96 kHz) are dominated by snapping shrimp, which increase their acoustic activity at night during the summer. Fish choruses, below 2 kHz, characterize the soundscape at sunset during the summer. Because there are 13 vessel passages per hour on average, causing acoustic interference with fish choruses 46% of the time, this MPA cannot be considered to be protected from noise. On the basis of the high seasonal variability of the soundscape components, this study proposes a one-year acoustic monitoring protocol using the soundscape methodology approach and discusses the concept of MPA size.

Mediterranean circulation perturbations over the last five centuries: Relevance to past Eastern Mediterranean Transient-type events.

The Eastern Mediterranean Transient (EMT) occurred in the Aegean Sea from 1988 to 1995 and is the most significant intermediate-to-deep Mediterranean overturning perturbation reported by instrumental records. The EMT was likely caused by accumulation of high salinity waters in the Levantine and enhanced heat loss in the Aegean Sea, coupled with surface water freshening in the Sicily Channel. It is still unknown whether similar transients occurred in the past and, if so, what their forcing processes were. In this study, sediments from the Sicily Channel document surface water freshening (SCFR) at 1910 ± 12, 1812 ± 18, 1725 ± 25 and 1580 ± 30 CE. A regional ocean hindcast links SCFR to enhanced deep-water production and in turn to strengthened Mediterranean thermohaline circulation. Independent evidence collected in the Aegean Sea supports this reconstruction, showing that enhanced bottom water ventilation in the Eastern Mediterranean was associated with each SCFR event. Comparison between the records and multi-decadal atmospheric circulation patterns and climatic external forcings indicates that Mediterranean circulation destabilisation occurs during positive North Atlantic Oscillation (NAO) and negative Atlantic Multidecadal Oscillation (AMO) phases, reduced solar activity and strong tropical volcanic eruptions. They may have recurrently produced favourable deep-water formation conditions, both increasing salinity and reducing temperature on multi-decadal time scales.

Are mussels able to distinguish underwater sounds? Assessment of the reactions of Mytilus galloprovincialis after exposure to lab-generated acoustic signals.

This study examined the effects of lab-generated acoustic signals on the behaviour and biochemistry of Mediterranean mussels (Mytilus galloprovincialis). The experiment was carried out in a tank equipped with a video-recording system using six groups of five mussels exposed to five acoustic treatments (each treatment was replicated three times) for 30min. The acoustic signals, with a maximum sound pressure level of 150dB rms re 1µPa, differed in frequency range as follows: low (0.1-5kHz), mid-low (5-10kHz), mid (10-20kHz), mid-high (20-40kHz) and high (40-60kHz). The exposure to sweeps did not produce any significant changes in the mussels' behaviour. Conversely, the specimens exposed to the low frequency band treatment showed significantly higher values of the following biochemical stress parameters measured in their plasma and tissues: glucose, total proteins, total haemocyte number (THC), heat shock protein 70 (Hsp70) expression, and Acetylcholinesterase (AChE) activity. The responses observed in the mussels exposed to low frequency sweeps enable us to suppose a biological and ecological role for this sound, which contains the main frequencies produced by both shipping traffic and the acoustic emissions of fish.

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.