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.

Acoustic recordings of rough-toothed dolphin (Steno bredanensis) offshore Eastern Sicily (Mediterranean Sea).

Rough-toothed dolphin's abundance and distribution is largely unknown worldwide and evaluation of its conservation status in the Mediterranean Sea is necessary. A rough-toothed dolphin was sighted offshore Eastern Sicily (Mediterranean Sea) in July 2017 and acoustic data were acquired in the same area of Watkins, Tyack, Moore, and Notarbartolo di Sciara [(1987). Mar. Mamm. Sci. 3, 78-82]. An automatic detection algorithm was developed to identify the echolocation clicks recorded within both datasets and a recurrent inter-click interval value was identified during the new encounter. Distinctive whistle classes were also identified with similar contour shapes within both datasets.

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.

Vessel noise pollution as a human threat to fish: assessment of the stress response in gilthead sea bream (Sparus aurata, Linnaeus 1758).

This study examined the effects of boat noise pollution on the stress indices of gilthead sea bream (Sparus aurata, Linnaeus 1758). To assess the stress response in these fish, biometric values and plasma parameters such as ACTH, cortisol, glucose, lactate, haematocrit, Hsp70, total protein, cholesterol, triglycerides and osmolarity were analysed. After acclimatization of the animals, the experiment was carried out in a tank fitted with underwater speakers where the fish were exposed to sound treatments (in duplicate) consisting of: 10 days of no sound (control treatment; the animals were only exposed to the experimental tank's background noise) and 10 days of noise derived from original recordings of motor boats, including recreational boats, hydrofoil, fishing boat and ferry boat (vessel noise treatment). The exposure to noise produced significant variations in almost all the plasma parameters assessed, but no differences were observed in weights and fork lengths. A PERMANOVA analysis highlighted significantly increased values (p < 0.05) of ACTH, cortisol, glucose, lactate, haematocrit, Hsp70, cholesterol, triglycerides and osmolarity in the fish exposed to vessel noise for 10 days. This study clearly highlights that anthropogenic noise negatively affects fish, and they are valuable targets for detailed investigations into the effects of this global pollutant. Finally, these experimental studies could represent part of the science that is able to improve the quality of the policies related to management plans for maritime spaces (Marine Strategy Framework Directive 56/2008 CE) that are aimed at stemming this pollutant phenomenon.

Assessment and Bioaccumulation of Heavy Metals in Water, Fish (wild and Farmed) and Associated Human Health Risk.

Worldwide, anthropogenic activities are causing the natural environment and aquaculture systems to become heavily contaminated with heavy metals, which can lead to consumer's health problems. In the current study, wild and farmed fish (n = 30) and water samples (n = 6) have been collected from the Chashma barrage and fish farm to assess the heavy metals concentration, i.e., Cu, Cd, Pb, Zn and Cr, in the water and some important organs (gills, liver, muscle, brain and bones) of wild and farmed fish (Labeo rohita) using Graphite furnace Atomic absorption spectrometry. Bioaccumulation factor and human health risk assessment were calculated to measure the health status of both fish and humans. Results show that in wild and farm fish's gills, muscles and bones, the trend of the heavy metals was Zn > Pb > Cu > Cd > Cr. On the other hand, the brain and liver show Zn > Cu > Pb > Cd > Cr trend. Comparatively, the heavy metals concentration was mostly higher (P < 0.05) in wild fish. Further; in both fish habitats (water) the heavy metals (Cd and Pb) concentration was higher than the WHO standard level, while in the body, Cd was higher (P < 0.05) in all studied organs except the muscle, Cr was only lower (P > 0.05) in muscle and brain. Pb was higher (P < 0.05) in all studied organs of both fish. Bioaccumulation of heavy metals was mostly higher (P < 0.05) in wild fish than in farmed. EDI and THQ were higher in wild fish, but the HI value was lower than 1 for both fish. Moreover, the PCA analysis suggests a positive correlation between heavy metals concentration in fish organs and the water of both fish (wild and farmed). Results indicated that farmed fish showed less potential risk to humans than wild fish.

Physiological and agonistic behavioural response of Procambarus clarkii to an acoustic stimulus.

This study examined the effects of an acoustic stimulus on the haemolymph and agonistic behaviour of the red swamp crayfish, Procambarus clarkii. The experiment was conducted in a tank equipped with a video recording system using six groups (three control and three test groups) of five adult crayfish (30 specimens in total). After 1 h of habituation, the behaviour of the crayfish was monitored for 2 h. During the second hour, the animals in the test groups were exposed to a linear sweep (frequency range 0.1-25 kHz; peak amplitude 148 dB(rms) re. 1 µPa at 12 kHz) acoustic stimulus for 30 min. Exposure to the noise produced significant variations in haemato-immunological parameters as well as a reduction in agonistic behaviour.

The underwater acoustic activities of the red swamp crayfish Procambarus clarkii.

This study describes the underwater acoustic behavior of the red swamp crayfish Procambarus clarkii. The study was conducted both in a tank and in the natural environment. The tank was equipped with video and acoustic recording systems. Observations were conducted to identify the underwater acoustic signals produced and their association with behavioral events and the movement status of the animals. In a lake in a natural reserve, a remote acoustic recording station was used to study the circadian underwater acoustic activity of the crayfish and to assess the acoustic features of the signals. The red swamp crayfish produces irregular trains of wide-band pulses (duration 0.4 ms, SPL(PK) 128 dB re 1 µPa, peak frequency 28 kHz, bandwidth(RMS) 20 kHz). The production of signals is positively related to intraspecific interactions (encounter/approach, fighting and successive Tail Flips). In the natural environment, acoustic activity is almost absent during the day, increases abruptly at sunset and continues until dawn. This study reveals the previously unknown underwater acoustic signals of Procambarus clarkii and the potential of passive acoustic methods to monitor the presence, the abundance and the behavioral activities of this invasive species.

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.

Effect of fortified feed with olive leaves extract on the haematological and biochemical parameters of Oreochromis niloticus (Nile tilapia).

Olive leaves have been used traditionally to improve the immunity against several infections. Current study aimed to examine the effect of fortified feed with olive leaves extract on haematological and biochemical parameters of Oreochromis niloticus. Total 400 healthy fish were divided randomly into four groups and were cultured for two months in glass tanks of 250 L water capacity in triplicate. Fortified feed was prepared with various extract, 1%, 1.5% and 2% concentration in diet, while, control-group diet was without extraction. Results showed that all the haematological parameters including WBC, RBC, Hb, PCV and plasma protein were significantly increased while the biochemical parameters such as globulin albumen ratio increased in fish which fed on 1% of extract, but the level of cholesterol and triglycerides was decreasing by increasing the concentration of extract. It was concluded that olive leaves extract can be used to enhance the immunity, growth and health of fish.

Biological Sound vs. Anthropogenic Noise: Assessment of Behavioural Changes in Scyliorhinus canicula Exposed to Boats Noise.

Despite the growing interest in human-made noise effects on marine wildlife, few studies have investigated the potential role of underwater noise on elasmobranch species. In this study, twelve specimens of small-spotted catshark (Scyliorhinus canicula) were exposed to biological and anthropogenic sounds in order to assess their behavioural changes in response to prey acoustic stimuli and to different amplitude levels of shipping noise. The sharks, individually held in aquariums, were exposed to four experimental acoustic conditions characterized by different spectral (Hz) components and amplitude (dB re 1 µPa) levels. The swimming behaviour and spatial distribution of sharks were observed. The results highlighted significant differences in swimming time and in the spatial use of the aquarium among the experimental conditions. When the amplitude levels of biological sources were higher than those of anthropogenic sources, the sharks' swimming behaviour was concentrated in the bottom sections of the aquarium; when the amplitude levels of anthropogenic sources were higher than biological ones, the specimens increased the time spent swimming. Moreover, their spatial distribution highlighted a tendency to occupy the least noisy sections of the aquarium. In conclusion, this study highlighted that anthropogenic noise is able to affect behaviour of catshark specimens and the impact depends on acoustic amplitude levels.

Bottlenose Dolphin (Tursiops truncatus) Whistle Modulation during a Trawl Bycatch Event in the Adriatic Sea.

Marine mammal vocal elements have been investigated for decades to assess whether they correlate with stress levels or stress indicators. Due to their acoustic plasticity, the interpretation of dolphins' acoustic signals of has been studied most extensively. This work describes the acoustic parameters detected in whistle spectral contours, collected using passive acoustic monitoring (PAM), in a bycatch event that involved three Bottlenose dolphins during midwater commercial trawling. The results indicate a total number of 23 upsweep whistles recorded during the bycatch event, that were analyzed based on the acoustic parameters as follows: (Median; 25th percentile; 75th percentile) Dr (second), total duration (1.09; 0.88; 1.24); fmin (HZ), minimum frequency (5836.4; 5635.3; 5967.1); fmax (HZ), maximum frequency, (11,610 ± 11,293; 11,810); fc (HZ), central frequency; (8665.2; 8492.9; 8982.8); BW (HZ), bandwidth (5836.4; 5635.3; 5967.1); Step, number of step (5; 4; 6). Furthermore, our data show that vocal production during the capture event was characterized by an undescribed to date combination of two signals, an ascending whistle (upsweep), and a pulsed signal that we called "low-frequency signal" in the frequency band between 4.5 and 7 kHz. This capture event reveals a novel aspect of T. truncatus acoustic communication, it confirms their acoustic plasticity, and suggests that states of discomfort are conveyed through their acoustic repertoire.

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.

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.

Behavioural and biochemical stress responses of Palinurus elephas after exposure to boat noise pollution in tank.

This study examined the effects of boat noise on the behavioural and biochemical parameters of the Mediterranean spiny lobster (Palinurus elephas). The experiment was conducted in a tank equipped with a video and audio recording system. 18 experimental trials, assigned to boat noise and control conditions, were performed using lobsters in single and group of 4 specimens. After a 1h habituation period, we audio- and video-recorded the lobsters for 1h. During the experimental phase, the animals assigned to the boat groups were exposed to boat noise pollution (a random sequence of boat noises). Exposure to the noise produced significant variations in locomotor behaviours and haemolymphatic parameters. Our results indicate that the lobsters exposed to boat noises increased significantly their locomotor activities and haemolymphatic bioindicator of stressful conditions such as glucose, total proteins, Hsp70 expression and THC when tested both singly and in groups.

Impact of an acoustic stimulus on the motility and blood parameters of European sea bass (Dicentrarchus labrax L.) and gilthead sea bream (Sparus aurata L.).

The physiological responses of fish to underwater noise are poorly understood and further information is needed to evaluate any possible negative effects of sound exposure. We exposed European sea bass and gilthead sea bream to a 0.1-1 kHz linear sweep (150 dB(rms) re 1 microPa). This band frequency is perceptible by many species of fish and is mainly produced by vessel traffic. We assessed the noise-induced motility reaction (analysing the movements) and the haematological responses (measuring blood glucose and lactate, and haematocrit levels). The noise exposure produced a significant increase in motility as well as an increase in lactate and haematocrit levels in sea bream and sea bass. A significant decrease of glucose was only observed in sea bream. A linear correlation between blood parameters and motility in fish exposed to the noise was observed. The acoustic stimulus produced intense muscle activity.