The Biological and Anthropogenic Soundscape of an Urbanized Port - the Charleston Harbor Estuary, South Carolina, USA.

Soundscape ecology provides a long-term, noninvasive approach to track animal behavior, habitat quality, and community structure over temporal and spatial scales. Using soniferous species as an indicator, biological soundscapes provide information about species and ecosystem health as well as their response and resiliency to potential stressors such as noise pollution. Charleston Harbor, South Carolina, USA provides important estuarine habitat for an abundance of marine life and is one of the busiest and fastest growing container ports in the southeast USA. Six passive acoustic recorders were deployed in the Charleston Harbor from December 2017 to June 2019 to determine biological patterns and human-associated influences on the soundscape. Anthropogenic noise was detected frequently across the estuary, especially along the shipping channel. Despite this anthropogenic noise, biological sound patterns were identified including snapping shrimp snaps (Alpheus spp. and Synalpheus spp.), fish calling and chorusing (Sciaenidae and Batrachoididae families), and bottlenose dolphin vocalizations. Biological response to anthropogenic activity varied among trophic levels, with decreased detection of fish calling when anthropogenic noise occurred and increased dolphin vocalizations in the presence of anthropogenic noise. Statistically, fine-scale, temporal patterns in biological sound were not clearly identified by sound pressure levels (SPLs), until files with anthropogenic noise presence were removed. These findings indicate that SPL patterns may be limited in their interpretation of biological activity for noisy regions and that the overall acoustic signature that we find in more pristine estuaries is lost in Charleston Harbor.

Sound pressure and particle motion components of the snaps produced by two snapping shrimp species (Alpheus heterochaelis and Alpheus angulosus).

Snapping shrimps are pervasive generators of underwater sound in temperate and tropical coastal seas across oceans of the world. Shrimp snaps can act as signals to conspecifics and provide acoustic information to other species and even to humans for habitat monitoring. Despite this, there are few controlled measurements of the acoustic parameters of these abundant acoustic stimuli. Here, the characteristics of snaps produced by 35 individuals of two species, Alpheus heterochaelis and Alpheus angulosus, are examined to evaluate the variability within and between the species. Animals were collected from the wild and the sound pressure and particle acceleration were measured at 0.2, 0.5, and 1 m from individual shrimp in controlled laboratory conditions to address the snap properties at communication-relevant distances. The source and sound exposure levels (at 1 m) were not significantly different between these two species. The frequency spectra were broadband with peak frequencies consistently below 10 kHz. The particle acceleration, the sound component likely detectable by shrimp, was measured across three axes. The directional amplitude variation suggests that the particle motion of snaps could act as a localization cue. The amplitudes of the snap pressure and acceleration decreased with distance, yet the levels remained sufficient for the predicted detection range by nearby conspecifics.

A changing estuary: Understanding historical patterns in salinity and fecal coliform levels in the May River, SC.

The May River, South Carolina watershed has undergone rapid increases in population and development from 1999 to 2017. This study aimed to understand the factors that influence salinity and fecal coliform levels in this estuary and how these levels changed from 1999 to 2017. This analysis revealed that salinity levels decreased in the headwaters, while variability increased. Additionally, fecal coliform increased from 1999 to 2017 throughout the hydrological network, with drastic changes occurring in the headwaters. Salinity and fecal coliform were influenced by spatial (distance from the mouth of the river), temporal (year, season, and tidal cycles), environmental (El Niño Southern Oscillation and rainfall), and anthropogenic parameters (population). This analysis suggests that the synergistic nature of climate change, resulting in more intense and frequent El Niño events, and watershed development may lead to further decreases in salinity and increases in fecal coliform levels in the May River estuary.

What's all that racket! Soundscapes, phenology, and biodiversity in estuaries.

There is now clear evidence that climate change affects terrestrial and marine ecosystems and can cause phenological shifts in behavior. Utilizing sound to demonstrate phenology is gaining popularity in terrestrial environments. In marine ecosystems, this technique is yet to be used due to a lack of multiyear datasets. Our study demonstrates soundscape phenology in an estuary using a six-year dataset. In this study, we showed that an increase in acoustic activity of snapping shrimp and certain fish species occurred earlier in years with warmer springs. In addition, we combined passive acoustics and traditional sampling methods (seines) and detected positive relationships between temporal patterns of the soundscape and biodiversity. This study shows that passive acoustics can provide information on the ecological response of estuaries to climate variability.

Automatic detection, classification, and quantification of sciaenid fish calls in an estuarine soundscape in the Southeast United States.

In the Southeast USA, major contributors to estuarine soundscapes are the courtship calls produced by fish species belonging to the family Sciaenidae. Long-term monitoring of sciaenid courtship sounds may be valuable in understanding reproductive phenology, but this approach produces massive acoustic datasets. With this in mind, we designed a feature-based, signal detector for sciaenid fish calls and tested the efficacy of this detector against manually reviewed data. Acoustic recorders were deployed to collect sound samples for 2 min every 20 min at four stations in the May River estuary, South Carolina, USA from February to November, 2014. Manual analysis of acoustic files revealed that four fish species, belonging to the family Sciaenidae, were the major sound producers in this estuarine soundscape, and included black drum (Pogonias cromis), silver perch (Bairdiella chrysoura), spotted seatrout (Cynoscion nebulosus), and red drum (Sciaenops ocellatus). Recorded calls served as an acoustic library of signature features that were used to create a signal detector to automatically detect, classify, and quantify the number of calls in each acoustic file. Correlation between manual and automatic detection was significant and precision varied from 61% to 100%. Automatic detection provided quantitative data on calling rates for this long-term data set. Positive temperature anomalies increased calling rates of black drum, silver perch, and spotted seatrout, while negative anomalies increased calling rates of red drum. Acoustic monitoring combined with automatic detection could be an additional or alternative method for monitoring sciaenid spawning and changes in phenology associated with climate change.

Boat noise in an estuarine soundscape - A potential risk on the acoustic communication and reproduction of soniferous fish in the May River, South Carolina.

The impact of boat related noise on marine life is a subject of concern, particularly for fish species that utilize acoustic communication for spawning purposes. The goal of this study was to quantify and examine the risk of boat noise on fish acoustic communication by performing acoustic monitoring of the May River, South Carolina (USA) from February to November 2013 using DSG-Ocean recorders. The number of boats detected increased from the source to the mouth with the highest detections near the Intracoastal Waterway (ICW). Boat noise frequency ranges overlapped with courtship sounds of silver perch (Bairdiella chrysoura), black drum (Pogonias cromis), oyster toadfish (Opsanus tau), spotted seatrout (Cynoscion nebulosus), and red drum (Sciaenops ocellatus). In the May River estuary, red drum may experience the greatest risk of auditory masking because of late afternoon choruses (21% time overlap with boat noise) and only one spawning location near the noisy ICW.

Acoustic monitoring indicates a correlation between calling and spawning in captive spotted seatrout (Cynoscion nebulosus).

BACKGROUND: Fish sound production is widespread throughout many families. Territorial displays and courtship are the most common reasons for fish sound production. Yet, there is still some questions on how acoustic signaling and reproduction are correlated in many sound-producing species. In the present study, our aim was to determine if a quantitative relationship exists between calling and egg deposition in captive spotted seatrout (Cynoscion nebulosus). This type of data is essential if passive acoustics is to be used to identify spawning aggregations over large spatial scales and monitor reproductive activity over annual and decadal timeframes. METHODS: Acoustic recorders (i.e., DSG-Oceans) were placed in three laboratory tanks to record underwater sound over an entire, simulated reproductive season. We enumerated the number of calls, calculated the received sound pressure level, and counted the number of eggs every morning in each tank. RESULTS: Spotted seatrout produced three distinct call types characterized as "drums," "grunts," and "staccatos." Spotted seatrout calling increased as the light cycle shifted from 13.5 to 14.5 h of light, and the temperature increased to 27.7 °C. Calling decreased once the temperature fell below 27.7 °C, and the light cycle shifted to 12 h of light. These temperature and light patterns followed the natural reproductive season observed in wild spotted seatrout in the Southeast United States. Spotted seatrout exhibited daily rhythms in calling. Acoustic signaling began once the lights turned off, and calling reached maximum activity approximately 3 h later. Eggs were released only on evenings in which spotted seatrout were calling. In all tanks, spotted seatrout were more likely to spawn when male fish called more frequently. A positive relationship between SPL and the number of eggs collected was found in Tanks 1 and 3. DISCUSSION: Our findings indicate that acoustic metrics can predict spawning potential. These findings are important because plankton tows may not accurately reflect spawning locations since egg capture is likely affected by predator activity and water currents. Instead, passive acoustics could be used to monitor spotted seatrout reproduction. Future studies can use this captive study as a model to record the estuarine soundscape precisely over long time periods to better understand how human-made stressors (e.g., climate change, noise pollution, and chemical pollutants) may affect spawning patterns.

Live CT imaging of sound reception anatomy and hearing measurements in the pygmy killer whale, Feresa attenuata.

In June 2008, two pygmy killer whales (Feresa attenuata) were stranded alive near Boca Grande, FL, USA, and were taken into rehabilitation. We used this opportunity to learn about the peripheral anatomy of the auditory system and hearing sensitivity of these rare toothed whales. Three-dimensional (3-D) reconstructions of head structures from X-ray computed tomography (CT) images revealed mandibles that were hollow, lacked a bony lamina medial to the pan bone and contained mandibular fat bodies that extended caudally and abutted the tympanoperiotic complex. Using auditory evoked potential (AEP) procedures, the modulation rate transfer function was determined. Maximum evoked potential responses occurred at modulation frequencies of 500 and 1000 Hz. The AEP-derived audiograms were U-shaped. The lowest hearing thresholds occurred between 20 and 60 kHz, with the best hearing sensitivity at 40 kHz. The auditory brainstem response (ABR) was composed of seven waves and resembled the ABR of the bottlenose and common dolphins. By changing electrode locations, creating 3-D reconstructions of the brain from CT images and measuring the amplitude of the ABR waves, we provided evidence that the neuroanatomical sources of ABR waves I, IV and VI were the auditory nerve, inferior colliculus and the medial geniculate body, respectively. The combination of AEP testing and CT imaging provided a new synthesis of methods for studying the auditory system of cetaceans.

Magnetic resonance imaging quality and volumes of brain structures from live and postmortem imaging of California sea lions with clinical signs of domoic acid toxicosis.

Our goal in this study was to compare magnetic resonance images and volumes of brain structures obtained alive versus postmortem of California sea lions Zalophus californianus exhibiting clinical signs of domoic acid (DA) toxicosis and those exhibiting normal behavior. Proton density-(PD) and T2-weighted images of postmortem-intact brains, up to 48 h after death, provided similar quality to images acquired from live sea lions. Volumes of gray matter (GM) and white matter (WM) of the cerebral hemispheres were similar to volumes calculated from images acquired when the sea lions were alive. However, cerebrospinal fluid (CSF) volumes decreased due to leakage. Hippocampal volumes from postmortem-intact images were useful for diagnosing unilateral and bilateral atrophy, consequences of DA toxicosis. These volumes were similar to the volumes in the live sea lion studies, up to 48 h postmortem. Imaging formalin-fixed brains provided some information on brain structure; however, images of the hippocampus and surrounding structures were of poorer quality compared to the images acquired alive and postmortem-intact. Despite these issues, volumes of cerebral GM and WM, as well as the hippocampus, were similar to volumes calculated from images of live sea lions and sufficient to diagnose hippocampal atrophy. Thus, postmortem MRI scanning (either intact or formalin-fixed) with volumetric analysis can be used to investigate the acute, chronic and possible developmental effects of DA on the brain of California sea lions.

Brominated flame retardants and organochlorine contaminants in winter flounder, harp and hooded seals, and North Atlantic right whales from the Northwest Atlantic Ocean.

Various brominated flame retardants (BFRs), including polybrominated diphenyl ethers (PBDEs) and current-use, non-PBDE BFRs, as well as organochlorine (OC) pesticides and polychlorinated biphenyls (PCBs), were measured in winter flounder, harp and hooded seals, and North Atlantic right whales from the Eastern United States and Canada. The concentrations of PBDEs in winter flounder and right whales were similar in magnitude to the levels of PCBs, which was unlike the pattern observed in seals. In these marine mammals, the levels of PBDEs were orders of magnitude lower than the levels of OCs and PCBs detected. Evidence existed for the accumulation of methoxylated (MeO)-PBDEs of natural origin in seals and right whales. Current-use, non-PBDE BFRs (including hexabromocyclododecane, pentabromoethylbenzene, hexabromobenzene, and pentabromotoluene) were detected in winter flounder and marine mammals. Future research should focus on monitoring PBDEs, current-use, non-PBDE BFRs, and MeO-BDEs of natural origin in marine organisms from Massachusetts and Cape Cod Bays.

Ultrasound detection in the Gulf menhaden requires gas-filled bullae and an intact lateral line.

Clupeiform fish species, including the Gulf menhaden (Brevoortia patronus) that belong to the subfamily Alosinae, can detect ultrasound. Clupeiform fishes are unique in that they have specialized gas-filled bullae in the head associated with the ear via the bulla membrane and with the lateral line via the lateral recess membrane. It has been hypothesized that the utricle of the inner ear is responsible for ultrasound detection through a specialized connection to the gas-filled bullae complex. Here, we show that the lateral line and its connection to the gas-filled bullae complex via the lateral recess are involved in ultrasound detection in Gulf menhaden. Removal of a small portion of the lateral line overlying the lateral recess membrane eliminates the ability of Gulf menhaden to detect ultrasound. We further show that the gas-filled bullae vibrates in response to ultrasound, that the gas-filled bullae are necessary for detecting ultrasound, and that the bullae connections to the lateral line via the lateral recess membrane play an important role in ultrasound detection. These results add a new dimension to the role of the lateral line and bullae as part of the ultrasonic detection system in Gulf menhaden.

Neuroanatomy and volumes of brain structures of a live California sea lion (Zalophus californianus) from magnetic resonance images.

The California sea lion (Zalophus californianus) has been a focal point for sensory, communication, cognition, and neurological disease studies in marine mammals. However, as a scientific community, we lack a noninvasive approach to investigate the anatomy and size of brain structures in this species and other free-ranging, live marine mammals. In this article, we provide the first anatomically labeled, magnetic resonance imaging-based atlas derived from a live marine mammal, the California sea lion. The brain of the California seal lion contained more secondary gyri and sulci than the brains of terrestrial carnivores. The olfactory bulb was present but small. The hippocampus of the California sea lion was found mostly in the ventral position with very little extension dorsally, quite unlike the canids and the mustelids, in which the hippocampus is present in the ventral position but extends dorsally above the thalamus. In contrast to the canids and the mustelids, the pineal gland of the California sea lion was strikingly large. In addition, we report three-dimensional reconstructions and volumes of cerebrospinal fluid, cerebral ventricles, total white matter (WM), total gray matter (GM), cerebral hemispheres (WM and GM), cerebellum and brainstem combined (WM and GM), and hippocampal structures all derived from magnetic resonance images. These measurements are the first to be determined for any pinniped species. In California sea lions, this approach can be used not only to relate cognitive and sensory capabilities to brain size but also to investigate the neurological effects of exposure to neurotoxins such as domoic acid.

Organohalogen contaminants and metabolites in cerebrospinal fluid and cerebellum gray matter in short-beaked common dolphins and Atlantic white-sided dolphins from the western North Atlantic.

Concentrations of several congeners and classes of organohalogen contaminants (OHCs) and/or their metabolites, namely organochlorine pesticides (OCs), polychlorinated biphenyls (PCBs), hydroxylated-PCBs (OH-PCBs), methylsulfonyl-PCBs (MeSO(2)-PCBs), polybrominated diphenyl ether (PBDE) flame retardants, and OH-PBDEs, were measured in cerebrospinal fluid (CSF) of short-beaked common dolphins (n = 2), Atlantic white-sided dolphins (n = 8), and gray seal (n = 1) from the western North Atlantic. In three Atlantic white-sided dolphins, cerebellum gray matter (GM) was also analyzed. The levels of OCs, PCBs, MeSO(2)-PCBs, PBDEs, and OH-PBDEs in cerebellum GM were higher than the concentrations in CSF. 4-OH-2,3,3',4',5-pentachlorobiphenyl (4-OH-CB107) was the only detectable OH-PCB congener present in CSF. The sum (Sigma) OH-PCBs/Sigma PCB concentration ratio in CSF was approximately two to three orders of magnitude greater than the ratio in cerebellum GM for dolphins.

Volumetric neuroimaging of the atlantic white-sided dolphin (Lagenorhynchus acutus) brain from in situ magnetic resonance images.

The structure and development of the brain are extremely difficult to study in free-ranging marine mammals. Here, we report measurements of total white matter (WM), total gray matter (GM), cerebellum (WM and GM), hippocampus, and corpus callosum made from magnetic resonance (MR) images of fresh, postmortem brains of the Atlantic white-sided dolphin (Lagenorhynchus acutus) imaged in situ (i.e., the brain intact within the skull, with the head still attached to the body). WM:GM volume ratios of the entire brain increased from fetus to adult, illustrating the increase in myelination during ontogeny. The cerebellum (WM and GM combined) of subadult and adult dolphins ranged from 13.8 to 15.0% of total brain size, much larger than that of primates. The corpus callosum mid-sagittal area to brain mass ratios (CCA/BM) ranged from 0.088 to 0.137, smaller than in most mammals. Dolphin hippocampal volumes were smaller than those of carnivores, ungulates, and humans, consistent with previous qualitative results assessed from histological studies of the bottlenose dolphin brain. These quantitative measurements of white matter, gray matter, corpus callosum, and hippocampus are the first to be determined from MR images for any cetacean species. We establish here an approach for accurately determining the size of brain structures from in situ MR images of stranded, dead dolphins. This approach can be used not only for comparative and developmental studies of marine mammal brains but also for investigation of the potential impacts of natural and anthropogenic chemicals on neurodevelopment and neuroanatomy in exposed marine mammal populations.

Cytochrome P4501A1 expression, polychlorinated biphenyls and hydroxylated metabolites, and adipocyte size of bottlenose dolphins from the Southeast United States.

Persistent organic pollutants (POPs) bioaccumulate in blubber of marine mammals. Therefore, it is important to understand the structure and dynamics of blubber layers and how they affect the accumulation of POPs and subsequent biochemical responses. We used established histological and immunohistochemical methods to document the structure of bottlenose dolphin (Tursiops truncatus) blubber and to assess the expression of cytochrome P4501A1 (CYP1A1) in skin-blubber biopsies of dolphins sampled in the waters off Charleston, SC (CHS) (N=38), and Indian River Lagoon, FL (IRL) (N=36). CYP1A1 expression was strongest and most frequent in capillary endothelial cells and was stratified in blubber; the greatest CYP1A1 staining was in the deepest layer. CYP1A1 expression in deep blubber and 2,3,7,8-TCDD Toxic Equivalents measured in the entire blubber were significantly higher in dolphins from CHS as compared to those from IRL. Adipocyte size was associated with the extent of CYP1A1 expression. Male dolphins with smaller adipocytes from CHS and IRL had higher levels of CYP1A1 expression in deep blubber. In CHS females, CYP1A1 expression in vascular endothelial cells varied with reproductive status. CYP1A1 expression in the deep layer was highest in simultaneously pregnant-lactating dolphins, and these dolphins had the smallest adipocytes in deep blubber. In all dolphins, CYP1A1 expression in the deep blubber layer was positively related to concentrations of hydroxylated PCBs (OH-PCBs) in plasma. In summary, redistribution of AHR agonists from blubber into the circulatory system may enhance PCB metabolism and production of OH-PCBs by induction of CYP1A1 in hepatocytes and, possibly, by induction of CYP1A1 in endothelial cells of the deep blubber. The OH-PCBs thus formed have the potential to interfere with thyroid hormone homeostasis.

Blubber morphology in wild bottlenose dolphins (Tursiops truncatus) from the Southeastern United States: influence of geographic location, age class, and reproductive state.

This study investigated blubber morphology and correlations of histological measurements with ontogeny, geography, and reproductive state in live, wild bottlenose dolphins (Tursiops truncatus) from the southeastern United States. Surgical skin-blubber biopsies (N=74) were collected from dolphins during capture-release studies conducted in two geographic locations: Charleston, SC (N=38) and Indian River Lagoon, FL (N=36). Histological analysis of blubber revealed stratification into superficial, middle, and deep layers. Adipocytes of the middle blubber were 1.6x larger in Charleston subadults than in Indian River Lagoon subadults (4,590+/-340 compared to 2,833+/-335 microm2 per cell). Charleston subadult dolphins contained higher levels of total blubber lipids than Charleston adult animals (49.3%+/-1.9% compared to 34.2%+/-1.7%), and this difference was manifested in more adipocytes in the middle blubber layer (19.2+/-0.9 compared to 14.9+/-0.5 cells per field). However, dolphins from Indian River Lagoon did not exhibit this pattern, and the adipocyte cell counts of subadults were approximately equal to those of the adults (16.0+/-1.4 compared to 13.4+/-0.8 cells per field). The colder year-round water temperatures in Charleston compared to Indian River Lagoon may explain these differences. Adipocytes in the deep blubber layer were significantly smaller in lactating and simultaneously pregnant and lactating animals compared to pregnant dolphins (840+/-179, 627+/-333, and 2,776+/-586 microm2 per cell, respectively). Total blubber lipid content and adipocyte size in the deep blubber of mothers with calves decreased linearly with calf length. Lactating females may utilize lipids from the deep blubber during periods of increased energetic demands associated with offspring care. This study demonstrates that ontogeny, geography, and reproductive state may influence morphological parameters such as structural fiber densities and adipocyte numbers and sizes, measured in bottlenose dolphin blubber.

Neuroanatomy of the subadult and fetal brain of the Atlantic white-sided dolphin (Lagenorhynchus acutus) from in situ magnetic resonance images.

This article provides the first anatomically labeled, magnetic resonance imaging (MRI) -based atlas of the subadult and fetal Atlantic white-sided dolphin (Lagenorhynchus acutus) brain. It differs from previous MRI-based atlases of cetaceans in that it was created from images of fresh, postmortem brains in situ rather than extracted, formalin-fixed brains. The in situ images displayed the classic hallmarks of odontocete brains: fore-shortened orbital lobes and pronounced temporal width. Olfactory structures were absent and auditory regions (e.g., temporal lobes and inferior colliculi) were enlarged. In the subadult and fetal postmortem MRI scans, the hippocampus was identifiable, despite the relatively small size of this structure in cetaceans. The white matter tracts of the fetal hindbrain and cerebellum were pronounced, but in the telencephalon, the white matter tracts were much less distinct, consistent with less myelin. The white matter tracts of the auditory pathways in the fetal brains were myelinated, as shown by the T2 hypointensity signals for the inferior colliculus, cochlear nuclei, and trapezoid bodies. This finding is consistent with hearing and auditory processing regions maturing in utero in L. acutus, as has been observed for most mammals. In situ MRI scanning of fresh, postmortem specimens can be used not only to study the evolution and developmental patterns of cetacean brains but also to investigate the impacts of natural toxins (such as domoic acid), anthropogenic chemicals (such as polychlorinated biphenyls, polybrominated diphenyl ethers, and their hydroxylated metabolites), biological agents (parasites), and noise on the central nervous system of marine mammal species.