Acoustic fish community in the Madeira Archipelago (North Atlantic Ocean): Characterization of sound diversity and daily patterns.

Marine ecosystems are increasingly subjected to anthropogenic pressures, which demands urgent monitoring plans. Understanding soundscapes can offer unique insights into the ocean status providing important information and revealing different sounds and their sources. Fishes can be prominent soundscape contributors, making passive acoustic monitoring (PAM) a potential tool to detect the presence of vocal fish species and to monitor changes in biodiversity. The major goal of this research was to provide a first reference of the marine soundscapes of the Madeira Archipelago focusing on fish sounds, as a basis for a long-term PAM program. Based on the literature, 102 potentially vocal and 35 vocal fish species were identified. Additionally 43 putative fish sound types were detected in audio recordings from two marine protected areas (MPAs) in the Archipelago: the Garajau MPA and the Desertas MPA. The Garajau MPA exhibited higher fish vocal activity, a greater variety of putative fish sound types and higher fish sound diversity. Lower abundance of sounds was found at night at both MPAs. Acoustic activity revealed a clear distinction between diurnal and nocturnal fish groups and demonstrated daily patterns of fish sound activity, suggesting temporal and spectral partitioning of the acoustic space. Pomacentridae species were proposed as candidates for some of the dominant sound types detected during the day, while scorpionfishes (Scorpaena spp.) were proposed as sources for some of the dominant nocturnal fish sounds. This study provides an important baseline about this community acoustic behaviour and is a valuable steppingstone for future non-invasive and cost-effective monitoring programs in Madeira.

Diagnosis and management of placentitis with severe funisitis in a multiparous Warmblood mare.

Placentitis is an important cause of reproductive losses in the equine industry. Many cases of clinical placentitis are not diagnosed until late in the course of the disease, and for this reason there is variability in the timing of intervention, diagnostic measurements, and treatment protocols. An 8-year-old multiparous Dutch Warmblood mare that was recently exposed to EHV-1 in the herd of origin presented for routine foaling management. Placentitis was diagnosed upon intake, and medical treatment was initiated. The mare delivered a term foal, and diagnostics for infectious etiology were unrewarding. While there were obvious clinical signs supporting a diagnosis of placentitis, histopathologic examination did not reveal active inflammatory lesions in the chorioallantois; however, severe funisitis was present. This report reviews early diagnosis and management of placentitis, demonstrates an array of classic and subtle pathologic lesions seen on microscopic examination, and discusses pathophysiology of placentitis.

Characterization of the vocal behavior of the miniature and transparent fish model, Danionella cerebruma).

Danionella cerebrum has recently been proposed as a promising model to investigate the structure and function of the adult vertebrate brain, including the development of vocal-auditory neural pathways. This genetically tractable and transparent cypriniform is highly vocal, but limited information is available on its acoustic behavior and underlying biological function. Our main goal was to characterize the acoustic repertoire and diel variation in sound production of D. cerebrum, as well as to investigate the relationship between vocal behavior and reproduction. Sound recordings demonstrated high vocal activity, with sounds varying from short sequences of pulses known as "bursts" (comprising up to 15 pulses) to notably longer sounds, termed "long bursts", which extended up to 349 pulses with over 2.7 s. Vocal activity peaked at midday and it was very low at night with only a few bursts. While the number of pulses was higher during the daytime, the interpulse interval was longer at night. In addition, calling time was positively associated with the number of viable eggs, suggesting that acoustic communication is important for reproduction. These preliminary findings reveal the potential of using D. cerebrum to investigate vocal plasticity and the implications for sexual selection and reproduction in a novel vertebrate model for neuroscience.

The role of acoustic signals in fish reproductiona).

This paper outlines my research path over three decades while providing a review on the role of fish sounds in mate choice and reproduction. It also intends to provide advice to young scientists and point toward future avenues in this field of research. An overview of studies on different fish model species shows that male mating acoustic signals can inform females and male competitors about their size (dominant frequency, amplitude, and sound pulse rate modulation), body condition (calling activity and sound pulse rate), and readiness to mate (calling rate, number of pulses in a sound). At least in species with parental care, such as toadfishes, gobies, and pomacentrids, calling activity seems to be the main driver of reproductive success. Playback experiments ran on a restricted number of species consistently revealed that females prefer vocal to silent males and select for higher calling rates. This personal synthesis concludes with the suggestion to increase knowledge on fish mating signals, especially considering the emerging use of fish sounds to monitor aquatic environments due to increasing threats, like noise pollution.

Correction: Vocal repertoire and consistency of call features in the meagre Argyrosomous regius (Asso, 1801).

[This corrects the article DOI: 10.1371/journal.pone.0241792.].

Mating sounds in the two-spotted goby, Pomatoschistus flavescens: Effects of water temperature on acoustic featuresa).

Acoustic signals in teleost fishes play a fundamental role in reproduction. As fish are ectothermic animals, temperature has the potential to change their signal production and detection, with further implications for mating interactions. In this study, we describe the mating sounds made by the two-spotted goby, Pomatoschistus flavescens, for the first time and further investigate the effect of temperature on the acoustic features. Courtship sounds of 15 two-spotted goby males were recorded at three different temperatures: 16 °C, 19 °C, and 21 °C. As seen for other marine gobies, two-spotted goby produced two courtship sounds: drums and thumps. Drums showed similar acoustic features to other Pomatoschistus species already studied. Calling rates for both kinds of sound were not affected by the increases in temperature. However, pulse rate increased from 16 °C to 19 °C and stabilised between 19 °C and 21 °C, suggesting that two-spotted gobies reached their physiological limits at 19 °C. Spectral features were also affected by temperature, presenting higher values at 19 °C. Whether or not the observed changes in acoustic features with temperature lead to changes in mating remains to be addressed. Studies like the present one are fundamental to better comprehend how reproduction will be affected by global warming in soniferous fishes.

Effects of temperature on acoustic and visual courtship and reproductive success in the two-spotted goby Pomatoschistus flavescens.

Fish are ectothermic and small changes in water temperature could greatly affect reproduction. The two-spotted goby is a small semi-pelagic species that uses visual and acoustic displays to mate. Here, we studied the effect of temperature (16 and 20 °C) on acoustic and visual courtship and associated reproductive success in 39 males. Temperature influenced male visual courtship performed outside the nest, but it did not influence calling rate and the number of laid eggs. Interestingly, the number of sounds (drums) was the sole predictor of spawning success. These findings suggest that exposure to different temperatures within the species' natural range affect courtship behaviour but not its reproductive success. We propose that finding the link between acoustic behaviour and reproduction in fishes offers the opportunity to monitor fish sounds both in the lab and in nature to learn how they respond to environmental changes and human impacts, namely global warming.

Lower low density lipoprotein cholesterol associates to higher mortality in non-diabetic heart failure patients.

Background: In patients with established heart failure (HF) low total cholesterol levels associate with worse prognosis. Evidence concerning the impact of Low-density lipoprotein cholesterol (LDL-c) in HF is scarce. We aimed to evaluate the prognostic impact of LDL-c in patients with HF, both with and without diabetes mellitus (DM). Methods: We retrospectively analyzed outpatients with chronic HF with systolic dysfunction followed in our HF clinic from January/2012 to May/2018. LDL-c was calculated using the Friedewald's formula. Patients without a complete lipid profile were excluded. The endpoint under analysis was all-cause mortality. Patients were followed until January/2021. A Cox-regression analysis was used to study the prognostic impact of LDL-c. The LDL-c cut-off used was 100 mg/dL (mean value). Analysis was stratified according to the coexistence of DM. Multivariate models were built adjusting for age, sex, coronary artery disease, atherosclerotic non-coronary artery disease, arterial hypertension, smoking status, statin use, severity of systolic dysfunction, creatinine clearance and evidence-based therapy. Results: We studied 522 chronic HF patients, mean age was 70 years, 66.5% males. Severe systolic dysfunction was present in 42.7%, 30.5% had coronary heart disease, 60.5% had arterial hypertension, 41.6% had DM. A total of 92.0% were treated with beta blocker, 87.5% with an ACEi/ARB and 29.1% with a MRA. During a median follow-up of 53 (interquartile range 33-73) months, 235 (45%) patients died. Patients with LDL-c ≤100 mg/dL presented increased multivariate-adjusted risk of all-cause mortality: HR = 1.58 (95% CI: 1.08-2.30), p = 0.02. When patients were stratified according to DM, LDL-c ≤100 mg/dL was independently associated with increased death risk - HR = 1.55 (95% CI:1.05-2.30), p = 0.03 in patients without DM; in patients with DM no association was detected - multivariate-adjusted HR = 1.18 (95% CI: 0.77-1.80), p = 0.44. Conclusion: Non-DM HF patients with LDL-c>100 mg/dL have a 35% reduction in the mortality risk when compared with those with lower values. The "cholesterol paradox" in HF also applies to LDL-c in non-DM patients.

Detection of invasive fish species with passive acoustics: Discriminating between native and non-indigenous sciaenids.

Invasive alien species have been rising exponentially in the last decades impacting biodiversity and ecosystem functioning. The soniferous weakfish, Cynoscion regalis, is a recent invasive sciaenid species in the Iberian Peninsula and was first reported in the Tagus estuary in 2015. There is concern about its possible impacts on native species, namely the confamiliar meagre, Argyrosomus regius, as there is overlap in their feeding regime, habitat use, and breeding behaviour. Here, we characterised the sciaenid-like sounds recently recorded in the Tagus estuary and showed that they are made by weakfish as they have similar numbers of pulses and pulse periods to the sounds made by captive breeding weakfish. We further demonstrate that breeding grunts from weakfish and the native sciaenid, recorded either in captivity or Tagus estuary, differ markedly in sound duration, number of pulses and pulse period in the two species, but overlap in their spectral features. Importantly, these differences are easily detected through visual and aural inspections of the recordings, making acoustic recognition easy even for the non-trained person. We propose that passive acoustic monitoring can be a cost-effective tool for in situ mapping of weakfish outside its natural distribution and an invaluable tool for early detection and to monitor its expansion.

Impact of anthropogenic noise on the survival and development of meagre (Argyrosomus regius) early life stages.

The growth of human populations has been driving an unprecedent and widespread increase in marine traffic, posing a real threat to marine biodiversity. Even though we are now aware of the negative effects of shipping noise exposure on fish, information about the impact on their early life stages continues to lack. Meagre (Argyrosomus regius) is a vocal fish that uses estuaries with high levels of anthropogenic noise pollution as both breeding areas and nurseries. Here, the effects of boat noise exposure on the development and survival of meagre larvae were studied. Embryos and larvae were exposed to either noise (boat noise playback) or control treatments (coils producing a similar electric field to the speakers) and hatching rate, survival rate, morphometric traits and stress-related biomarkers, at hatching and at 2 days-post-hatching (dph) were analyzed. Results showed no conclusive effects of the impact of boat noise playback, even though there was an increased lipid droplet consumption and a decrease in body depth at 2dph larvae under this stressor. The assessment of oxidative stress and energy metabolism-related biomarkers at hatching showed a marginal decrease in superoxide dismutase (SOD) activity and no changes in DNA damage or electron transport system activity (ETS), although it cannot be disregarded that those effects could only be visible at later stages of larval development. Whether these morphological and developmental results have implications in later stages remains to be investigated. Further studies with longer exposure and wild meagre could help deepen this knowledge and provide a better understanding of how anthropogenic noise can impact meagre early stages.

On the importance of longer-term exposure to stressors - A critical review and proposal for multigenerational testing in standard soil invertebrates.

Standard laboratory tests to describe the impact of stressors (most notably: chemicals) on organisms offer a good compromise between feasibility and outcome, i.e., they should be reproducible and provide robust results. However, these tests may underestimate the potential effects of prolonged exposures, particularly for persistent contaminants. Within the last years, we have observed an increase in studies aiming to target prolonged exposure, e.g., via an extended test duration or by multigenerational (MG) exposure. Seemingly, both reduced and increased impacts have been observed in these studies, but it is also clear that no unique test setup was used, and test designs vary widely among studies. To better describe long term effects, MG is a highly relevant aspect which deserves more consideration at various testing and assessment levels. Therefore, we conducted a literature review focusing on available studies performed with soil invertebrates, exposed to stressors for periods longer than in standard laboratory tests, i.e., full life cycle tests, as well as extensions to standard and MG tests. So far, it has been recommended that such studies should cover more than one generation, but this statement is probably too vague. In this contribution, we summarize and critically discuss the information provided in the literature, and we provide suggestions for future research. The currently available test results from long-term studies have produced clear evidence to recommend the implementation of long-term tests in existing regulatory testing requirements (e.g., for pesticides), in particular for persistent substances and also for delayed effects. Consequently, we recommend the inclusion of such longer exposure test designs (e.g., as annexes) in current OECD and ISO guidelines. However, when doing so, the long-term test designs proposed so far have to be critically adapted for a selected set of representative soil invertebrate test species.

Antibacterial and hemostatic capacities of cellulose nanocrystalline-reinforced poly(vinyl alcohol) electrospun mats doped with Tiger 17 and pexiganan peptides for prospective wound healing applications.

Infection is a major issue in chronic wound care. Different dressings have been developed to prevent microbial propagation, but an effective, all-in-one (cytocompatible, antimicrobial and promoter of healing) solution is still to be uncovered. In this research, polyvinyl alcohol (PVA) nanofibrous mats reinforced with cellulose nanocrystal (CNC), at 10 and 20% v/v ratios, were produced by electrospinning, crosslinked with glutaraldehyde vapor and doped with specialized peptides. Crosslinking increased the mats' fiber diameters but maintained their bead-free morphology. Miscibility between polymers was confirmed by Fourier-transform infrared spectroscopy and thermal evaluations. Despite the incorporation of CNC having reduced the mats' mechanical performance, it improved the mats' surface energy and its structural stability over time. Pexiganan with an extra cysteine group was functionalized onto the mats via hydroxyl- polyethylene glycol 2-maleimide, while Tiger 17 was physisorbed to preserve its cyclic conformation. Antimicrobial assessments demonstrated the peptide-doped mat's effectiveness against Staphylococcus aureus and Pseudomonas aeruginosa; pexiganan contributed mostly for such outcome. Tiger 17 showed excellent capacity in accelerating clotting. Cytocompatibility evaluations attested to these mats' safety. C90/10 PVA/CNC mats were deemed the most effective from the tested group and, thus, a potentially effective option for chronic wound treatments.

On virus and nanomaterials - Lessons learned from the innate immune system - ACE activation in the invertebrate model Enchytraeus crypticus.

Current human research on COVID-19 - SARS-CoV-2 (Severe Acute Respiratory Syndrome-Corona Virus) showed that ACE2 (Angiotensin Converting Enzyme 2) is a functional receptor to which the spike proteins attach. Invertebrates have been exposed to a wide array of threats for millennia and their immune system has evolved to deal with these efficiently. The annelid Enchytraeus crypticus, a standard ecotoxicological species, is an invertebrate species where extensive mechanisms of response studies are available, covering all levels from gene to population responses. Nanomaterials (NMs) are often perceived as invaders (e.g. virus) and can enter the cell covered by a corona, triggering similar responses. We created a database on E. crypticus ACE gene expression, aiming to analyse the potential knowledge transfer between invertebrates and vertebrates. Total exposure experiments sum 87 stress conditions for 18 different nanomaterials (NMs). ACE expression following TiO2 NM exposure was clearly different from other NMs showing a clear (6-7 fold) ACE down-regulation, not observed for any other NMs. Other NMs, notably Ag NMs, and to some extent Cu NMs, caused ACE up-regulation (up to 4 fold). The extensive knowledge from response to NMs can support the immuno-research community, especially to develop therapies for virus that trigger the innate immune system.

Full life cycle test with Eisenia fetida - copper oxide NM toxicity assessment.

Copper oxide nanomaterials (CuONM) are widely used, e.g. as antimicrobial coatings, wood preservatives,fertilizers, etc. Life cycle aspects of Copper Nanomaterials (CuONM) toxicity have been scarcely studied in earthworms, as the focus has been on standard survival and reproduction toxicity tests. Standard tests do not allow for an understanding of which life cycle stage is the most sensitive, and how this can be impacted by CuONMs toxicity to cause longer term population level effects. Since CuONM may release free Cu ions (Cu2+) it is relevant to compare CuONM toxicity with CuCl2 salt. Therefore, the aim of the present study was to assess the effects of CuONM and CuCl2 throughout the different stages of the full life cycle (FLC) of Eisenia fetida while comparing it with the OECD standard test. Additional endpoints included juvenile survival, growth, maturation, besides survival and reproduction. The FLC test showed that e.g. juvenile survival was especially susceptible during the first 28 days post-hatching, neither juvenile growth nor time to maturity were related to exposure concentration. Both CuONM and CuCl2 caused a concentration-dependent decrease in population growth, while a hormesis effect was observed at low concentrations of CuCl2. A reduction in instantaneous growth rate was observed at higher concentrations. The FLC test and the OECD test had different exposure history therefore the ECx values are not comparable across the test systems. Hence, the FLC test enabled the detection of the most vulnerable developmental stages and elucidate different life stage sensitivities between the two Cu exposures.

Tiger 17 and pexiganan as antimicrobial and hemostatic boosters of cellulose acetate-containing poly(vinyl alcohol) electrospun mats for potential wound care purposes.

In this research, we propose to engineer a nanostructured mat that can simultaneously kill bacteria and promote an environment conducive to healing for prospective wound care. Polyvinyl alcohol (PVA) and cellulose acetate (CA) were combined at different polymer ratios (100/0, 90/10, 80/20% v/v), electrospun and crosslinked with glutaraldehyde vapor. Crosslinked fibers increased in diameter (from 194 to 278 nm), retaining their uniform structure. Fourier-transform infrared spectroscopy and thermal analyses proved the excellent miscibility between polymers. CA incorporation incremented the fibers swelling capacity and reduced the water vapor and air permeabilities of the mats, preventing the excessive drying of wounds. The antimicrobial peptide cys-pexiganan and the immunoregulatory peptide Tiger 17 were incorporated onto the mats via polyethylene glycol spacer (hydroxyl-PEG2-maleimide) and physisorbed, respectively. Time-kill kinetics evaluations revealed the mats effectiveness against Staphylococcus aureus and Pseudomonas aeruginosa. Tiger 17 played a major role in accelerating clotting of re-calcified plasma. Data reports for the first time the collaborative effect of pexiganan and Tiger 17 against bacterial infections and in boosting hemostasis. Cytocompatibility data verified the peptide-modified mats safety. Croslinked 90/10 PVA/CA mats were deemed the most promising combination due to their moderate hydrophilicity and permeabilities, swelling capacity, and high yields of peptide loading.

Boat noise impacts Lusitanian toadfish breeding males and reproductive outcome.

Anthropogenic noise is a growing threat to marine organisms, including fish. Yet very few studies have addressed the impact of anthropogenic noise on fish reproduction, especially in situ. In this study, we investigated the impacts of boat noise exposure in the reproductive success of wild Lusitanian toadfish (Halobatrachus didactylus), a species that relies on advertisement calls for mate attraction, using behavioural, physiological and reproductive endpoints. Two sets of artificial nests were deployed in the Tagus estuary and exposed to either ambient sound or boat noise during their breeding season. Toadfish males spontaneously used these nests to breed. We inspected nests for occupation and the presence of eggs in six spring low tides (in two years) and assessed male vocal activity and stress responses. Boat noise did not affect nest occupation by males but impacted reproductive success by decreasing the likelihood of receiving eggs, decreasing the number of live eggs and increasing the number of dead eggs, compared to control males. Treatment males also showed depressed vocal activity and slightly higher cortisol levels. The assessment of oxidative stress and energy metabolism-related biomarkers revealed no oxidative damage in noise exposed males despite having lower antioxidant responses and pointed towards a decrease in the activity levels of energy metabolism-related biomarkers. These results suggest that males exposed to boat noise depressed their metabolism and their activity (such as parental care and mate attraction) to cope with an acoustic stressor, consistent with a freezing defensive response/behaviour. Together, our study demonstrates that boat noise has severe impacts on reproductive fitness in Lusitanian toadfish. We argue that, at least fishes that cannot easily avoid noise sources due to their dependence on specific spawning sites, may incur in significant direct fitness costs due to chronic noise exposure.

Nano-pesticides: the lunch-box principle-deadly goodies (semio-chemical functionalised nanoparticles that deliver pesticide only to target species).

Nature contains many examples of "fake promises" to attract "prey", e.g., predatory spiders that emit the same sex-attractant-signals as moths to catch them at close range and male spiders that make empty silk-wrapped gifts in order to mate with a female. Nano-pesticides should ideally mimic nature by luring a target and killing it without harming other organisms/species. Here, we present such an approach, called the lunch-box or deadly-goodies approach. The lunch-box consists of three main elements (1) the lure (semio-chemicals anchored on the box), (2) the box (palatable nano-carrier), and (3) the kill (advanced targeted pesticide). To implement this approach, one needs to draw on the vast amount of chemical ecological knowledge available, combine this with recent nanomaterial techniques, and use novel advanced pesticides. Precision nano-pesticides can increase crop protection and food production whilst lowering environmental impacts.

Boat noise impacts early life stages in the Lusitanian toadfish: A field experiment.

Marine traffic is the most common and chronic source of ocean noise pollution. Despite the evidence of detrimental effects of noise exposure on fish, knowledge about the effects on the critical early life stages - embryos and larvae - is still scarce. Here, we take a natural habitat-based approach to examine potential impacts of boat noise exposure in early life stages in a wild fish population of the Lusitanian toadfish (Halobatrachus didactylus). In-situ experiments were carried out in the Tagus estuary, an estuary with significant commercial and recreational boat traffic. Nests with eggs were exposed to either ambient (control) or boat noise (treatment), for 1 fortnight. Eggs were photographed before being assigned to each treatment, and after exposure, to count number of eggs and/or larvae to assess survival, and sampled to study development and oxidative stress and energy metabolism-related biomarkers. Data concerns 4 sampling periods (fortnights) from 2 years. Results indicate that offspring survival did not differ between treatments, but boat noise induced a detrimental effect on embryos and larvae stress response, and on larvae development. Embryos showed reduced levels of electron transport system (ETS), an energy metabolism-related biomarker, while larvae showed higher overall stress responses, with increased levels of superoxide dismutase (SOD) and DNA damage (oxidative stress related responses), ETS, and reduced growth. With this study, we provided the first evidence of detrimental effects of boat noise exposure on fish development in the field and on stress biomarker responses. If these critical early stages are not able to compensate and/or acclimate to the noise stress later in the ontogeny, then anthropogenic noise has the potential to severely affect this and likely other marine fishes, with further consequences for populations resilience and dynamics.

Impact of chromium on the soil invertebrate model Enchytraeus crypticus (Oligochaeta) in standard reproduction and full life cycle tests.

Chromium (Cr) has many applications in industry and day-to-day life, entering the terrestrial environment mostly from anthropogenic sources. Despite the fact that Cr is essential, it can be harmful in biota at high concentrations. Full life cycle tests (FLCt) are scarce, representing a gap for soil ecotoxicology. Hence, the effects of Cr were studied using the soil organism Enchytraeus crypticus in the 46-day FLCt and the standard 21-day enchytraeid reproduction test (ERT). FLCt hatching (day-11) and time to reach maturity (day 22-25) were the most sensitive endpoints, representing a delay, partly recovered with time. Reproduction was reduced to similar levels in both the ERT and the FLCt (EC50∼320 mg Cr/kg), but survival was more affected in the ERT (LC50 = 377 mg Cr/kg) than in the FLC (LC50 = 467 mg Cr/kg). This could be due to the induction of stress response mechanisms at earlier life stages (cocoons/juveniles), providing protection to toxicity in a later stage (adults). FLCt results provided considerably more details and data points, and the additional endpoints allowed to explain the source of observed effects, e.g. a direct impact on fecundity and not only due to adult mortality. The estimated population growth curves confirmed the significant effect of 320 mg Cr/kg and onwards, reflecting the impact of Cr on both cocoon production and juvenile performance, with follow-up consequences for reproduction. An Adverse Outcome Pathway was drafted. The FLCt design is recommended as an upgrade of the current standard ERT, which could be integrated into the existing OECD ERT test guideline.

Boat noise affects meagre (Argyrosomus regius) hearing and vocal behaviour.

Aquatic noise has increased in last decades imposing new constraints on aquatic animals' acoustic communication. Meagre (Argyrosomus regius) produce loud choruses during the breeding season, likely facilitating aggregations and mating, and are thus amenable to being impacted by anthropogenic noise. We assessed the impact of boat noise on this species acoustic communication by: evaluating possible masking effects of boat noise on hearing using Auditory Evoked Potentials (AEP) and inspecting changes in chorus sound levels from free ranging fish upon boat passages. Our results point to a significant masking effect of anthropogenic noise since we observed a reduction of ca. 20 dB on the ability to discriminate conspecific calls when exposed to boat noise. Furthermore, we verified a reduction in chorus energy during ferryboat passages, a behavioural effect that might ultimately impact spawning. This study is one of few addressing the effects of boat noise by combining different methodologies both in the lab and with free ranging animals.