Eric W. Montie

Hyperbaric computed tomographic measurement of lung compression in seals and dolphins.

SUMMARY Lung compression of vertebrates as they dive poses anatomical and physiological challenges. There has been little direct observation of this. A harbor and a gray seal, a common dolphin and a harbor porpoise were each imaged post mortem under pressure using a radiolucent, fiberglass, water-filled pressure vessel rated to a depth equivalent of 170 m. The vessel was scanned using computed tomography (CT), and supported by a rail and counterweighted carriage magnetically linked to the CT table movement. As pressure increased, total buoyancy of the animals decreased and lung tissue CT attenuation increased, consistent with compression of air within the lower respiratory tract. Three-dimensional reconstructions of the external surface of the porpoise chest showed a marked contraction of the chest wall. Estimation of the volumes of different body compartments in the head and chest showed static values for all compartments except the lung, which showed a pressure-related compression. The depth of estimated lung compression ranged from 58 m in the gray seal with lungs inflated to 50% total lung capacity (TLC) to 133 m in the harbor porpoise with lungs at 100% TLC. These observations provide evidence for the possible behavior of gas within the chest of a live, diving mammal. The estimated depths of full compression of the lungs exceeds previous indirect estimates of the depth at which gas exchange ceases, and concurs with pulmonary shunt measurements. If these results are representative for living animals, they might suggest a potential for decompression sickness in diving mammals.

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.

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. Anat Rec, 2009.

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

SUMMARY 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.

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.

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. Anat Rec, 291:263–282, 2008.

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.

Magnetic resonance imaging and volumetric analysis: Novel tools to study the effects of thyroid hormone disruption on white matter development

Humans and wildlife are exposed to environmental pollutants that have been shown to interfere with the thyroid hormone system and thus may affect brain development. Our goal was to expose pregnant rats to propylthiouracil (PTU) to measure the effects of a goitrogen on white matter development in offspring using magnetic resonance imaging (MRI) and volumetric analysis. We exposed pregnant Sprague Dawley (SD) rats to 3 or 10 ppm PTU from gestation day 7 (GD7) until postnatal day 25 (P25) to determine the effects on white matter (WM), gray matter (GM), and hippocampus volumes in offspring. We sacrificed offspring at P25 but continued the life of some offspring to P90 to measure persistent effects in adult animals. P25 offspring exposed to 10 ppm PTU displayed lowered levels of triiodothyronine (T3) and thyroxine (T4); cerebral WM, GM, and total brain volumes were significantly lower than the volumes in control animals. P90 adults exposed to 10 ppm PTU displayed normal T3 levels but lowered T4 levels; WM, GM, total brain, and hippocampal volumes were significantly lower than the volumes in control adults. Both P25 and P90 rats exposed to 10 ppm PTU displayed significant reductions in percent WM as well as heterotopias in the corpus callosum. Exposure to 3 ppm PTU did not produce any significant effects. These results suggest that MRI coupled with volumetric analysis is a powerful tool in assessing the effects of thyroid hormone disruption on white matter development and brain structure. This approach holds great promise in assessing neurotoxicity of xenobiotics in humans and wildlife.

Seasonal and Spatial Patterns of Fish Sound Production in the May River, South Carolina

AbstractMany fish species produce sounds that are associated with reproductive behavior, and hydrophone recordings can be used to document this behavior in soniferous fishes. We recorded the sounds produced by a fish community and determined general seasonal and spatial patterns of fish sound production in the May River, South Carolina, as a means to assess general temporal and spatial patterns of reproduction. Two-minute sound files, temperature, salinity, and depth were recorded at 27 stations at monthly intervals along the May River in Bluffton, South Carolina, from January 2012 to December 2012. We identified species-specific calls from Spotted Seatrout Cynoscion nebulosus, Oyster Toadfish Opsanus tau, Silver Perch Bairdiella chrysoura, Black Drum Pogonias cromis, and Red Drum Sciaenops ocellatus. General species-specific seasonal patterns were noted; some species produced sounds within a specific time period and temperature range. Seasonally, Silver Perch, Oyster Toadfish, and Black Drum began callin...

Influences of biological variables and geographic location on circulating concentrations of thyroid hormones in wild bottlenose dolphins (Tursiops truncatus).

Thyroid hormones (TH) are key regulators of metabolism and development, yet our understanding of the variability in serum TH concentrations in free-ranging marine mammals is limited. Thus, we examined the interrelationships between TH and age, sex, reproductive status, geographic location, and ocean temperatures in wild bottlenose dolphins (Tursiops truncatus). Circulating concentrations of TH (total thyroxine (tT(4)), free T(4) (fT(4)), and total triiodothyronine (tT(3))) were determined in a total of 195 dolphins; 80 from the coastal waters of Charleston, South Carolina (CHS) and 115 from the Indian River Lagoon, Florida (IRL). Age had the most influence on circulating TH concentrations in dolphins at both sites with decreasing concentrations (p<0.0001) observed with increasing age for all TH. No significant differences were found between males and non-reproductive females. Geographic location significantly influenced tT(4) and tT(3) concentrations; CHS dolphins had higher concentrations than IRL animals. These TH differences between CHS and IRL dolphins may be attributed to the colder year-round water temperature that CHS dolphins inhabit compared to IRL dolphins and could constitute an adaptive response to their colder environment. Results from this study highlight the importance of establishing reference values for dolphins in different geographic locations to support valid comparisons. This initial assessment provides a foundation of how biological and environmental variables could affect circulating TH in dolphins, which will help to elucidate the impacts of disease, pollution, and climate change on the thyroid hormone system of aquatic mammals.