Manatee cognition in the wild: an exploration of the manatee mind and behavior through neuroanatomy, psychophysics, and field observations.

Cognition refers to the mechanisms for acquiring, processing, storing, and acting on information, all of which are critical to understanding the behavior of animals. These mechanisms are poorly known in manatees, especially how they are expressed in the wild. To expand our understanding of manatee cognition, we gathered information from behavioral experimentation in the laboratory, neuroanatomical research, controlled field studies, integrated laboratory and field measurement, and natural history observations (published reports, written surveys, and interviews with knowledgeable observers). Laboratory research, both neuroanatomical and behavioral, provided the most empirical data, primarily on sensory/perceptual capacities. Inferences from these data and narratives from surveys and interviews illuminated possibilities for higher order cognition. Evidence from field measurements was sparse, although substantial amounts of information have been collected from tracking data and to a lesser extent vessel impact studies, which can be used to infer cognitive attributes. Manatees are tactile-auditory specialists with complementary visual and chemosensory abilities. They demonstrate learning characteristics typical of vertebrates. Movement tracking data plus direct observations suggest that they have good spatial cognition, indicated by their ability to traverse complicated water networks and memory for foraging and warm water sites. They engage in a wide range of play-like, object manipulation, and mimetic behaviors, which suggests cognitive capacities beyond basic associative learning. Understanding manatee cognition beyond the laboratory will be necessary for conservation of manatees as they face challenges such as habitat degradation and threats from water-borne vessel traffic. There is a clear need for more direct research in natural settings.

Detection of hydrodynamic stimuli by the postcranial body of Florida manatees (Trichechus manatus latirostris).

Manatees live in shallow, frequently turbid waters. The sensory means by which they navigate in these conditions are unknown. Poor visual acuity, lack of echolocation, and modest chemosensation suggest that other modalities play an important role. Rich innervation of sensory hairs that cover the entire body and enlarged somatosensory areas of the brain suggest that tactile senses are good candidates. Previous tests of detection of underwater vibratory stimuli indicated that they use passive movement of the hairs to detect particle displacements in the vicinity of a micron or less for frequencies from 10 to 150 Hz. In the current study, hydrodynamic stimuli were created by a sinusoidally oscillating sphere that generated a dipole field at frequencies from 5 to 150 Hz. Go/no-go tests of manatee postcranial mechanoreception of hydrodynamic stimuli indicated excellent sensitivity but about an order of magnitude less than the facial region. When the vibrissae were trimmed, detection thresholds were elevated, suggesting that the vibrissae were an important means by which detection occurred. Manatees were also highly accurate in two-choice directional discrimination: greater than 90% correct at all frequencies tested. We hypothesize that manatees utilize vibrissae as a three-dimensional array to detect and localize low-frequency hydrodynamic stimuli.

Identifying coagulopathies in the pathophysiology of cold stress syndrome in the Florida manatee Trichechus manatus latirostris.

Cold stress syndrome (CSS) in the Florida manatee Trichechus manatus latirostris has been defined as morbidity and mortality resulting from prolonged exposure to water temperatures <20°C. The pathophysiology is described as multifactorial, involving nutritional, immunological and metabolic disturbances; however, the exact mechanisms are unknown. We hypothesized that thromboembolic complications contribute to the pathophysiology of CSS in addition to the previously described factors. During the winter of 2014-2015, 10 Florida manatees with clinical signs of CSS were presented to Lowry Park Zoo, Tampa, FL, USA. Thromboelastography (TEG) and coagulation panels were performed at admission. In addition, coagulation panel data from 23 retrospective CSS cases were included in the analyses. There were numerous differences between mean values of TEG and coagulation parameters for healthy manatees and those for CSS cases. Among TEG parameters, reaction time (R), clot formation time (K) and percentage of clot lysed after 30 min (LY30) values were significantly different (p < 0.05) between the 2 groups. CSS cases also had significantly higher mean D-dimer concentration and coagulation factor XI activity, prolonged mean activated partial thromboplastin time (aPTT) and significantly decreased mean antithrombin activity. These combined abnormalities include clinicopathologic criteria of disseminated intravascular coagulation, indicating an increased risk of thromboembolic disease associated with manatee CSS.

PROPOSAL AND APPLICATION OF A NOVEL DISSEMINATED INTRAVASCULAR COAGULATION SCORING SYSTEM IN THE FLORIDA MANATEE (TRICHECHUS MANATUS LATIROSTRIS).

Disseminated intravascular coagulopathy (DIC) is an acquired disorder of hemostasis resulting in activation of the coagulation and fibrinolytic pathways. It is reported secondarily to multiple disease processes and can be associated with increased mortality. Previous research at Tampa's Lowry Park Zoo (LPZ) demonstrated that Florida manatees ( Trichechus manatus latirostris) with cold stress syndrome (CSS) demonstrated thromboembolic disease. The object of this retrospective study was to establish the presence and clinical relevance of DIC in Florida manatees admitted to LPZ for rehabilitation from 07 March 2010 to 15 August 2015. A coagulation panel, including prothrombin time, partial thromboplastin time, platelet count, fibrinogen level, and D-dimer level was used to diagnose DIC. There were 100 cases identified in the study period: 35 trauma, 43 CSS, 17 secondary to harmful algae blooms (HAB), and five miscellaneous. Manatees with CSS had the highest incidence of DIC with 24 of 43 cases (56%) affected, followed by trauma with 18 of 35 cases (52%) affected. None of the manatees with HAB were found to have DIC. Manatees that developed DIC during rehabilitation or when DIC progressed did not survive. Due to the clinical implications of DIC, identifying its presence and recognizing its severity could improve clinical outcomes by enabling more intensive treatment protocols.

Golgi Analysis of Neuron Morphology in the Presumptive Somatosensory Cortex and Visual Cortex of the Florida Manatee (Trichechus manatus latirostris).

The current study investigates neuron morphology in presumptive primary somatosensory (S1) and primary visual (V1) cortices of the Florida manatee (Trichechus manatus latirostris) as revealed by Golgi impregnation. Sirenians, including manatees, have an aquatic lifestyle, a large body size, and a relatively large lissencephalic brain. The present study examines neuron morphology in 3 cortical areas: in S1, dorsolateral cortex area 1 (DL1) and cluster cortex area 2 (CL2) and in V1, dorsolateral cortex area 4 (DL4). Neurons exhibited a variety of morphological types, with pyramidal neurons being the most common. The large variety of neuron types present in the manatee cortex was comparable to that seen in other eutherian mammals, except for rodents and primates, where pyramid-shaped neurons predominate. A comparison between pyramidal neurons in S1 and V1 indicated relatively greater dendritic branching in S1. Across all 3 areas, the dendritic arborization pattern of pyramidal neurons was also similar to that observed previously in the afrotherian rock hyrax, cetartiodactyls, opossums, and echidnas but did not resemble the widely bifurcated dendrites seen in the large-brained African elephant. Despite adaptations for an aquatic environment, manatees did not share specific neuron types such as tritufted and star-like neurons that have been found in cetaceans. Manatees exhibit an evolutionarily primitive pattern of cortical neuron morphology shared with most other mammals and do not appear to have neuronal specializations for an aquatic niche.

Eight-choice sound localization by manatees: performance abilities and head related transfer functions.

Two experiments investigated the ability and means by which two male Florida manatees (Trichechus manatus latirostris) may determine the direction of a sound source. An eight-choice discrimination paradigm was used to determine the subjects' sound localization abilities of five signal conditions covering a range of frequencies, durations, and levels. Subjects performed above the 12.5% chance level for all broadband frequencies and were able to localize sounds over a large level range. Errors were typically located to either side of the signal source location when presented in the front 180° but were more dispersed when presented from locations behind the subject. Front-to-back confusions were few and accuracy was greater when signals originated from the front 180°. Head-related transfer functions were measured to determine if frequencies were filtered by the manatee body to create frequency-specific interaural level differences (ILDs). ILDs were found for all frequencies as a function of source location, although they were largest with frequencies above 18 kHz and when signals originated to either side of the subjects. Larger ILDs were found when the signals originated behind the subjects. A shadowing-effect produced by the body may explain the relatively low occurrence of front-back confusions in the localization study.

Elaboration and Innervation of the Vibrissal System in the Rock Hyrax (Procavia capensis).

Mammalian tactile hairs are commonly found on specific, restricted regions of the body, but Florida manatees represent a unique exception, exhibiting follicle-sinus complexes (FSCs, also known as vibrissae or tactile hairs) on their entire body. The orders Sirenia (including manatees and dugongs) and Hyracoidea (hyraxes) are thought to have diverged approximately 60 million years ago, yet hyraxes are among the closest relatives to sirenians. We investigated the possibility that hyraxes, like manatees, are tactile specialists with vibrissae that cover the entire postfacial body. Previous studies suggested that rock hyraxes possess postfacial vibrissae in addition to pelage hair, but this observation was not verified through histological examination. Using a detailed immunohistochemical analysis, we characterized the gross morphology, innervation and mechanoreceptors present in FSCs sampled from facial and postfacial vibrissae body regions to determine that the long postfacial hairs on the hyrax body are in fact true vibrissae. The types and relative densities of mechanoreceptors associated with each FSC also appeared to be relatively consistent between facial and postfacial FSCs. The presence of vibrissae covering the hyrax body presumably facilitates navigation in the dark caves and rocky crevices of the hyrax's environment where visual cues are limited, and may alert the animal to predatory or conspecific threats approaching the body. Furthermore, the presence of vibrissae on the postfacial body in both manatees and hyraxes indicates that this distribution may represent the ancestral condition for the supraorder Paenungulata.

Neuron Types in the Presumptive Primary Somatosensory Cortex of the Florida Manatee (Trichechus manatus latirostris).

Within afrotherians, sirenians are unusual due to their aquatic lifestyle, large body size and relatively large lissencephalic brain. However, little is known about the neuron type distributions of the cerebral cortex in sirenians within the context of other afrotherians and aquatic mammals. The present study investigated two cortical regions, dorsolateral cortex area 1 (DL1) and cluster cortex area 2 (CL2), in the presumptive primary somatosensory cortex (S1) in Florida manatees (Trichechus manatus latirostris) to characterize cyto- and chemoarchitecture. The mean neuron density for both cortical regions was 35,617 neurons/mm(3) and fell within the 95% prediction intervals relative to brain mass based on a reference group of afrotherians and xenarthrans. Densities of inhibitory interneuron subtypes labeled against calcium-binding proteins and neuropeptide Y were relatively low compared to afrotherians and xenarthrans and also formed a small percentage of the overall population of inhibitory interneurons as revealed by GAD67 immunoreactivity. Nonphosphorylated neurofilament protein-immunoreactive (NPNFP-ir) neurons comprised a mean of 60% of neurons in layer V across DL1 and CL2. DL1 contained a higher percentage of NPNFP-ir neurons than CL2, although CL2 had a higher variety of morphological types. The mean percentage of NPNFP-ir neurons in the two regions of the presumptive S1 were low compared to other afrotherians and xenarthrans but were within the 95% prediction intervals relative to brain mass, and their morphologies were comparable to those found in other afrotherians and xenarthrans. Although this specific pattern of neuron types and densities sets the manatee apart from other afrotherians and xenarthrans, the manatee isocortex does not appear to be explicitly adapted for an aquatic habitat. Many of the features that are shared between manatees and cetaceans are also shared with a diverse array of terrestrial mammals and likely represent highly conserved neural features. A comparative study across manatees and dugongs is necessary to determine whether these traits are specific to one or more of the manatee species, or can be generalized to all sirenians.

Detection of hydrodynamic stimuli by the Florida manatee (Trichechus manatus latirostris).

Florida manatees inhabit the coastal and inland waters of the peninsular state. They have little difficulty navigating the turbid waterways, which often contain obstacles that they must circumnavigate. Anatomical and behavioral research suggests that the vibrissae and associated follicle-sinus complexes that manatees possess over their entire body form a sensory array system for detecting hydrodynamic stimuli analogous to the lateral line system of fish. This is consistent with data highlighting that manatees are tactile specialists, evidenced by their specialized facial morphology and use of their vibrissae during feeding and active investigation/manipulation of objects. Two Florida manatees were tested in a go/no-go procedure using a staircase method to assess their ability to detect low-frequency water movement. Hydrodynamic vibrations were created by a sinusoidally oscillating sphere that generated a dipole field at frequencies from 5 to 150 Hz, which are below the apparent functional hearing limit of the manatee. The manatees detected particle displacement of less than 1 µm for frequencies of 15-150 Hz and of less than a nanometer at 150 Hz. Restricting the facial vibrissae with various size mesh openings indicated that the specialized sensory hairs played an important role in the manatee's exquisite tactile sensitivity.

Audiogram and auditory critical ratios of two Florida manatees (Trichechus manatus latirostris).

Manatees inhabit turbid, shallow-water environments and have been shown to have poor visual acuity. Previous studies on hearing have demonstrated that manatees possess good hearing and sound localization abilities. The goals of this research were to determine the hearing abilities of two captive subjects and measure critical ratios to understand the capacity of manatees to detect tonal signals, such as manatee vocalizations, in the presence of noise. This study was also undertaken to better understand individual variability, which has been encountered during behavioral research with manatees. Two Florida manatees (Trichechus manatus latirostris) were tested in a go/no-go paradigm using a modified staircase method, with incorporated 'catch' trials at a 1:1 ratio, to assess their ability to detect single-frequency tonal stimuli. The behavioral audiograms indicated that the manatees' auditory frequency detection for tonal stimuli ranged from 0.25 to 90.5 kHz, with peak sensitivity extending from 8 to 32 kHz. Critical ratios, thresholds for tone detection in the presence of background masking noise, were determined with one-octave wide noise bands, 7-12 dB (spectrum level) above the thresholds determined for the audiogram under quiet conditions. Manatees appear to have quite low critical ratios, especially at 8 kHz, where the ratio was 18.3 dB for one manatee. This suggests that manatee hearing is sensitive in the presence of background noise and that they may have relatively narrow filters in the tested frequency range.

Parcellation in the dorsal column nuclei of Florida manatees (Trichechus manatus latirostris) and rock hyraxes (Procavia capensis) indicates the presence of body barrelettes.

Florida manatees (Trichechus manatus latirostris) and rock hyraxes (Procavia capensis) exhibit expanded tactile arrays of vibrissae that are distributed not only on the face but also on the entire postfacial body. In contrast, the vibrissae of most mammals are principally restricted to the face. These facial vibrissae may be associated with central nervous system representations known as barrels in the cerebral cortex, barreloids in the thalamus, and barrelettes in the trigeminal nuclei of the brainstem. To date, vibrissae representations found within the brainstem have been principally limited to facial vibrissae representations in the trigeminal nuclei. We hypothesized that the tactile specializations of the manatee and rock hyrax would produce a unique modification of typical mammalian central nervous system organization, with postfacial vibrissae representations appearing in the cuneate and gracile nuclei as "body barrelettes." Using histological and histochemical methods, including cresyl violet, myelin, and cytochrome oxidase processing, we first delineated the rostral, middle, and caudal zones of the cuneate and gracile nuclei. Within the middle zone, divisions were present, including extensive parcellation in the cluster region, particularly in manatees. These clusters were particularly densely distributed and distinguishable in the presumptive postfacial body representations in the cuneate and gracile nuclei but otherwise shared many attributes with the barrelettes found in the trigeminal nuclei of other species. This study represents the first characterization of postfacial body vibrissae representations, or "body barrelettes," in the brainstem of any species. Previous studies have predominantly focused on facial vibrissae representations, which have served for decades as a model for sensory organization and plasticity. Our results extend what is known about vibrissae representations in the central nervous system to include expansions related to peripheral specializations of the postfacial body. Unusual somatosensory adaptations in the manatee and rock hyrax are highly informative regarding how mammalian brain organization responds to evolutionary pressures on sensory systems.

Phrenic nerve afferent activation of neurons in the cat SI cerebral cortex.

Stimulation of respiratory afferents elicits neural activity in the somatosensory region of the cerebral cortex in humans and animals. Respiratory afferents have been stimulated with mechanical loads applied to breathing and electrical stimulation of respiratory nerves and muscles. It was hypothesized that stimulation of the phrenic nerve myelinated afferents will activate neurons in the 3a and 3b region of the somatosensory cortex. This was investigated in cats with electrical stimulation of the intrathoracic phrenic nerve and C(5) root of the phrenic nerve. The somatosensory cortical response to phrenic afferent stimulation was recorded from the cortical surface, contralateral to the phrenic nerve, ispilateral to the phrenic nerve and with microelectrodes inserted into the cortical site of the surface dipole. Short-latency, primary cortical evoked potentials (1 degrees CEP) were recorded with stimulation of myelinated afferents of the intrathoracic phrenic nerve in the contralateral post-cruciate gyrus of all animals (n = 42). The mean onset and peak latencies were 8.5 +/- 5.7 ms and 21.8 +/- 9.8 ms, respectively. The rostro-caudal surface location of the 1 degrees CEP was found between the rostral edge of the post-cruciate dimple (PCD) and the rostral edge of the ansate sulcus, medio-lateral location was between 2 mm lateral to the sagittal sulcus and the lateral end of the cruciate sulcus. Histological examination revealed that the 1 degrees CEP sites were recorded over areas 3a and 3b of the SI somatosensory cortex. Intracortical activation of 16 neurons with two patterns of neural activity was recorded: (1) short-latency, short-duration activation of neurons and (2) long-latency, long-duration activation of neurons. Short-latency neurons had a mean onset latency of 10.4 +/- 3.1 ms and mean burst duration of 10.1 +/- 3.2 ms. The short-latency units were recorded at an average depth of 1.7 +/- 0.5 mm below the cortical surface. The long-latency neurons had a mean onset latency of 36.0 +/- 4.2 ms and mean burst duration of 32.2 +/- 8.4 ms. The long-latency units were recorded at an average depth of 2.4 +/- 0.2 mm below the cortical surface. The results of the study demonstrated that phrenic nerve afferents have a short-latency central projection to the SI somatosensory cortex. The phrenic afferents activated neurons in lamina III and IV of areas 3a and 3b. The cortical representation of phrenic nerve afferents is medial to the forelimb, lateral to the hindlimb, similar to thoracic loci, hence the phrenic afferent SI site in the cat homunculus is consistent with body position (thoracic region) rather than spinal segment (C(5)-C(7)). The phrenic afferent activation of the somatosensory cortex is bilateral, with the ipsilateral cortical activation occurring subsequent to the contralateral. These results support the hypothesis that phrenic afferents provide somatosensory information to the cerebral cortex which can be used for diaphragmatic proprioception and somatosensation.

Developmental uterine anomalies in cats and dogs undergoing elective ovariohysterectomy.

OBJECTIVE: To describe the characteristics and frequency of gross uterine anomalies in cats and dogs undergoing elective ovariohysterectomy. DESIGN: Prospective and retrospective case series. ANIMALS: 53,258 cats and 32,660 dogs undergoing elective ovariohysterectomy at 26 clinics in the United States and Canada during 2007. PROCEDURES: Clinics prospectively reported gross anomalies and submitted tissues from abnormal reproductive tracts identified during surgery. Records from a feral cat spay-neuter clinic were evaluated retrospectively. RESULTS: Suspected congenital anomalies of the uterus were identified in 0.09% (49/53,258) of female cats and 0.05% (15/32,660) of female dogs. Uterine anomalies identified included unicornuate uterus (33 cats and 11 dogs), segmental agenesis of 1 uterine horn (15 cats and 3 dogs), and uterine horn hypoplasia (1 cat and 1 dog). Ipsilateral renal agenesis was present in 29.4% (10/34) of cats and 50.0% (6/12) of dogs with uterine anomalies in which kidneys were evaluated. Mummified ectopic fetuses were identified in 4 cats with uterine anomalies. Both ovaries and both uterine tubes were present in most animals with uterine anomalies. CONCLUSIONS AND CLINICAL RELEVANCE: Urogenital anomalies were twice as common in cats as in dogs. Identification of uterine developmental anomalies in dogs and cats should trigger evaluation of both kidneys and both ovaries because ipsilateral renal agenesis is common, but both ovaries are likely to be present and should be removed during ovariohysterectomy.

Posterior parietal cortex as part of a neural network for directed attention in rats.

A rodent model of directed attention has been developed based upon behavioral analysis of contralateral neglect, pharmacological manipulations, and anatomical analysis of neural circuitry. In each of these three domains the rodent model exhibits striking similarities to humans. We hypothesize that there is a specific thalamo-cortical-basal ganglia network that subserves spatial attentional functions. Key components of this network are medial agranular and posterior parietal cortex, dorsocentral striatum, and the lateral posterior thalamic nucleus. Several issues need to be addressed before we can hope to realistically understand or model the functions of this network. Among these are the roles of medial versus lateral posterior parietal cortex; cholinergic mechanisms in attention; interhemispheric interactions; the role of synchronous firing at the cortical, striatal, and thalamic levels; interactions between cortical and thalamic projections to the striatum; interactions between cortical and nigral inputs to the thalamus; the role of collicular inputs to the lateral posterior thalamic nucleus; the role of cerebral cortex versus superior colliculus in driving the motor output expressed as orienting behavior during directed attention; the extent to which the circuitry we describe for directed attention also plays a role in other forms of attention.

Nogo-A inhibition induces recovery from neglect in rats.

Neglect is a complex human cognitive spatial disorder typically induced by damage to prefrontal or posterior parietal association cortices. Behavioral treatments for neglect rarely generalize outside of the therapeutic context or across tasks within the same therapeutic context. Recovery, when it occurs, is spontaneous over the course of weeks to months, but often it is incomplete. A number of studies have indicated that anti-Nogo-A antibodies can be used to enhance plasticity and behavioral recovery following damage to motor cortex, and spinal cord. In the present studies the anti-Nogo-A antibodies IN-1, 7B12, or 11C7 were applied intraventricularly to adult rats demonstrating severe neglect produced by unilateral medial agranular cortex lesions in rats. The three separate anti-Nogo-A antibody groups were treated immediately following the medial agranular cortex lesions. Each of the three antibodies induced dramatic significant behavioral recovery from neglect relative to controls. Severing the corpus callosum to destroy inputs from the contralesional hemisphere resulted in reinstatement of severe neglect, pointing to a possible role of interhemispheric mechanisms in behavioral recovery from neglect.

Striatal projections from the rat lateral posterior thalamic nucleus.

The dorsocentral striatum (DCS) has been implicated as an associative striatal area receiving inputs from several cortical areas including medial agranular cortex (AGm), posterior parietal cortex (PPC), and visual association cortex to form a cortical-subcortical circuit involved in directed attention and neglect. The lateral posterior thalamic nucleus (LP) may also play a role in directed attention and neglect because LP has robust reciprocal connections with these cortical areas and projects to DCS. We used anterograde axonal tracing to map thalamostriatal projections from LP and surrounding thalamic nuclei, with a focus on projections to DCS. The thalamic nuclei investigated included LP, laterodorsal thalamic nucleus (LD), central lateral nucleus (CL), and posterior thalamic nucleus (Po). We found that the mediorostral part of LP (LPMR) projects strongly to DCS as well as to the dorsal peripheral region of the striatum. Further, there is topography within LPMR and DCS such that the far medial LPMR projects to the central region of DCS (projection area of AGm) and the central LPMR projects to the dorsal region of DCS (projection area of PPC and Oc2M). In contrast, the laterorostral part of LP (LPLR) and other thalamic nuclei surrounding LP project to dorsolateral to dorsomedial peripheral regions of the striatum but do not project to DCS. These findings indicate that DCS is a region of convergence for thalamostriatal and corticostriatal projections from regions that are themselves interconnected, serving as the key element of the corticostriatal-thalamic network mediating spatial processing and directed attention.

Quantification of synaptic density in corticostriatal projections from rat medial agranular cortex.

Medial agranular cortex (AGm) has a prominent bilateral projection to the dorsocentral striatum (DCS). We wished to develop a normal baseline by which to assess neuronal plasticity in this corticostriatal system in rats with neglect resulting from a unilateral lesion in AGm, followed by treatment with agents that promote sprouting and functional recovery in other systems. Injections of biotinylated dextran amine were made into AGm in normal rats, and unbiased sampling was used to quantify the density of axons and axonal varicosities present in DCS (the latter represent presynaptic profiles). Labeling density in contralateral DCS is approximately half of that seen in ipsilateral DCS (this ratio is 0.50 for axons, 0.55 for varicosities). The ratio of varicosities is stable over a greater than seven-fold range of absolute densities. There is no consistent relationship between the absolute density of axons and axon varicosities; however, the ratio measures are strongly correlated. We conclude that changes in the contralateral/ipsilateral ratio of axon density after experimental treatments do reflect changes in synaptic density, but axon varicosities are likely to be the most sensitive anatomical parameter by which to assess plasticity at the light microscopic level.

Thromboelastography in wild Florida manatees (Trichechus manatus latirostris).

BACKGROUND: Thromboelastography (TEG) provides a comprehensive evaluation of blood clot formation. This test can be used to identify abnormalities in coagulation by assessing multiple aspects of the clotting cascade, including the speed of clot initiation and formation, clot strength, and ultimately fibrinolysis. Thromboembolic disease has been hypothesized to play a role in the pathophysiology of cold stress syndrome (CSS), an important cause of mortality in the threatened Florida manatee (Trichechus manatus latirostris). OBJECTIVES: The objective of this study was to establish thromboelastography RIs using the TEG 5000 with citrated whole blood samples and kaolin activation in wild, healthy manatees. METHODS: In December 2014 and January 2015, 29 wild manatees (17 male and 12 female) were blood sampled as part of the annual wild manatee health assessments organized by United States Geological Survey (USGS). TEG was performed using citrated kaolin-activated samples. RESULTS: The samples were obtained from manatees caught in Crystal River, Citrus County, and used to identify the mean ± SD of normal TEG parameters: R = reaction time 2.1 minutes (0.8), K = clot formation time 0.8 min (0), α angle = 83.1° (2), MA = maximum amplitude 75 mm (7.6), and LY30 = clot lysis 0.41% (0.68). No significant differences were found between manatee size, sex, or time between sampling and running the test. CONCLUSIONS: Manatee TEG parameters demonstrate a relatively hypercoagulable condition when compared to other mammals. This information will facilitate detection of changes in hemostasis during injury and disease and provide a valuable reference range.

Adaptations in the structure and innervation of follicle-sinus complexes to an aquatic environment as seen in the Florida manatee (Trichechus manatus latirostris).

Florida manatees are large-bodied aquatic herbivores that use large tactile vibrissae for several purposes. Facial vibrissae are used to forage in a turbid water environment, and the largest perioral vibrissae can also grasp and manipulate objects. Other vibrissae distributed over the entire postfacial body appear to function as a lateral line system. All manatee vibrissae emanate from densely innervated follicle-sinus complexes (FSCs) like those in other mammals, although proportionately larger commensurate with the caliber of the vibrissae. As revealed by immunofluorescence, all manatee FSCs have many types of C, Adelta and Abeta innervation including Merkel, club, and longitudinal lanceolate endings at the level of the ring sinus, but they lack other types such as reticular and spiny endings at the level of the cavernous sinus. As in non-whisking terrestrial species, the inner conical bodies of facial FSCs are well innervated but lack Abeta-fiber terminals. Importantly, manatee FSCs have two unique types of Abeta-fiber endings. First, all of the FSCs have exceptionally large-caliber axons that branch to terminate as novel, gigantic spindle-like endings located at the upper ring sinus. Second, facial FSCs have smaller caliber Abeta fibers that terminate in the trabeculae of the cavernous sinus as an ending that resembles a Golgi tendon organ. In addition, the largest perioral vibrissae, which are used for grasping, have exceptionally well-developed medullary cores that have a structure and dense small-fiber innervation resembling that of tooth pulp. Other features of the epidermis and upper dermis structure and innervation differ from that seen in terrestrial mammals.

Topographical organization of the facial motor nucleus in Florida manatees (Trichechus manatus latirostris).

Florida manatees (Trichechus manatus latirostris) possess modified vibrissae that are used in conjunction with specialized perioral musculature to manipulate vegetation for ingestion, and aid in the tactile exploration of their environment. Therefore it is expected that manatees possess a large facial motor nucleus that exhibits a complex organization relative to other taxa. The topographical organization of the facial motor nucleus of five adult Florida manatees was analyzed using neuroanatomical methods. Cresyl violet and hematoxylin staining were used to localize the rostrocaudal extent of the facial motor nucleus as well as the organization and location of subdivisions within this nucleus. Differences in size, length, and organization of the facial motor nucleus among mammals correspond to the functional importance of the superficial facial muscles, including perioral musculature involved in the movement of mystacial vibrissae. The facial motor nucleus of Florida manatees was divided into seven subnuclei. The mean rostrocaudal length, width, and height of the entire Florida manatee facial motor nucleus was 6.6 mm (SD 8 0.51; range: 6.2-7.5 mm), 4.7 mm (SD 8 0.65; range: 4.0-5.6 mm), and 3.9 mm (SD 8 0.26; range: 3.5-4.2 mm), respectively. It is speculated that manatees could possess direct descending corticomotorneuron projections to the facial motornucleus. This conjecture is based on recent data for rodents, similiarities in the rodent and sirenian muscular-vibrissal complex, and the analogous nature of the sirenian cortical Rindenkerne system with the rodent barrel system.