Independent erosion of conserved transcription factor binding sites points to shared hindlimb, vision and external testes loss in different mammals.

Genetic variation in cis-regulatory elements is thought to be a major driving force in morphological and physiological changes. However, identifying transcription factor binding events that code for complex traits remains a challenge, motivating novel means of detecting putatively important binding events. Using a curated set of 1154 high-quality transcription factor motifs, we demonstrate that independently eroded binding sites are enriched for independently lost traits in three distinct pairs of placental mammals. We show that these independently eroded events pinpoint the loss of hindlimbs in dolphin and manatee, degradation of vision in naked mole-rat and star-nosed mole, and the loss of external testes in white rhinoceros and Weddell seal. We additionally show that our method may also be utilized with more than two species. Our study exhibits a novel methodology to detect cis-regulatory mutations which help explain a portion of the molecular mechanism underlying complex trait formation and loss.

Implications of Scientific Collaboration Networks on Studies of Aquatic Vertebrates in the Brazilian Amazon.

The quantity of wildlife extracted from the Amazon has increased in the past decades as a consequence of an increase in human population density and income growth. To evaluate the spatial distribution of studies on subsistence and/or commercial hunting conducted in the Brazilian Amazon, we selected eight mid-sized and large-bodied aquatic vertebrate species with a history of human exploitation in the region. We used a combination of searches in the gray and scientific literature from the past 24 years to provide an updated distributional map of studies on the target species. We calculated the distances between the study sites and the locations of the research institutes/universities that the first and last authors of the same study were affiliated to. For the period of 1990 to 2014, we found 105 studies on the subsistence and/or commercial hunting of aquatic vertebrates in the Brazilian Amazon in 271 locations that involved 43 institutions (37 Brazilian and 6 international). The spatial distribution of the studies across the Brazilian Amazon varied, but over 80% took place in the northeast and central Amazon, encompassing three States of the Legal Brazilian Amazon (Amazonas, 51.42%; Pará, 19.05%; and Amapá, 16.19%). Over half of the research study sites (52.91%) were within 500 km of the research institute/university of the first or last authors. Some research institutes/universities did not have any inter-institutional collaborations, while others collaborated with eight or more institutes. Some research institutes/universities conducted many studies, had an extensive collaboration network, and contributed greatly to the network of studies on Amazonian aquatic vertebrates. Our research contributes to the knowledge of studies on the subsistence and/or commercial hunting of the most exploited aquatic vertebrates of the Brazilian Amazon, illustrates the impact that collaboration networks have on research, and highlights potential areas for improvement and the generation of new collaborations.

Sublethal red tide toxin exposure in free-ranging manatees (Trichechus manatus) affects the immune system through reduced lymphocyte proliferation responses, inflammation, and oxidative stress.

The health of many Florida manatees (Trichechus manatus latirostris) is adversely affected by exposure to blooms of the toxic dinoflagellate, Karenia brevis. K. brevis blooms are common in manatee habitats of Florida's southwestern coast and produce a group of cyclic polyether toxins collectively referred to as red tide toxins, or brevetoxins. Although a large number of manatees exposed to significant levels of red tide toxins die, several manatees are rescued from sublethal exposure and are successfully treated and returned to the wild. Sublethal brevetoxin exposure may potentially impact the manatee immune system. Lymphocyte proliferative responses and a suite of immune function parameters in the plasma were used to evaluate effects of brevetoxin exposure on health of manatees rescued from natural exposure to red tide toxins in their habitat. Blood samples were collected from rescued manatees at Lowry Park Zoo in Tampa, FL and from healthy, unexposed manatees in Crystal River, FL. Peripheral blood leukocytes (PBL) isolated from whole blood were stimulated with T-cell mitogens, ConA and PHA. A suite of plasma parameters, including plasma protein electrophoresis profiles, lysozyme activity, superoxide dismutase (SOD) activity, and reactive oxygen/nitrogen (ROS/RNS) species, was also used to assess manatee health. Significant decreases (p<0.05) in lymphocyte proliferation were observed in ConA and PHA stimulated lymphocytes from rescued animals compared to non-exposed animals. Significant correlations were observed between oxidative stress markers (SOD, ROS/RNS) and plasma brevetoxin concentrations. Sublethal exposure to brevetoxins in the wild impacts some immune function components, and thus, overall health, in the Florida manatee.

Unique biochemical and mineral composition of whale ear bones.

Abstract Cetaceans are obligate aquatic mammals derived from terrestrial artiodactyls. The defining characteristic of cetaceans is a thick and dense lip (pachyosteosclerotic involucrum) of an ear bone (the tympanic). This unique feature is absent in modern terrestrial artiodactyls and is suggested to be important in underwater hearing. Here, we investigate the mineralogical and biochemical properties of the involucrum, as these may hold clues to the aquatic adaptations of cetaceans. We compared bioapatites (enamel, dentine, cementum, and skeletal bone) of cetaceans with those of terrestrial artiodactyls and pachyosteosclerotic ribs of manatees (Sirenia). We investigated organic, carbonate, and mineral composition as well as crystal size and crystallinity index. In all studied variables, bioapatites of the cetacean involucrum were intermediate in composition and structure between those of tooth enamel on the one hand and those of dentine, cementum, and skeletal bone on the other. We also studied the amino acid composition of the cetacean involucrum relative to that of other skeletal bone. The central involucrum had low glycine and hydroxyproline concentrations but high concentrations of nonessential amino acids, unlike most bone samples but similar to the tympanic of hippos and the (pachyosteosclerotic) ribs of manatees. These amino acid results are evidence of rapid bone development. We hypothesize that the mineralogical and amino acid composition of cetacean bullae differs from that of other bone because of (1) functional modifications for underwater sound reception and (2) structural adaptations related to rapid ossification.

Estimating upper bounds for occupancy and number of manatees in areas potentially affected by oil from the Deepwater Horizon oil spill.

The explosion of the Deepwater Horizon drilling platform created the largest marine oil spill in U.S. history. As part of the Natural Resource Damage Assessment process, we applied an innovative modeling approach to obtain upper estimates for occupancy and for number of manatees in areas potentially affected by the oil spill. Our data consisted of aerial survey counts in waters of the Florida Panhandle, Alabama and Mississippi. Our method, which uses a Bayesian approach, allows for the propagation of uncertainty associated with estimates from empirical data and from the published literature. We illustrate that it is possible to derive estimates of occupancy rate and upper estimates of the number of manatees present at the time of sampling, even when no manatees were observed in our sampled plots during surveys. We estimated that fewer than 2.4% of potentially affected manatee habitat in our Florida study area may have been occupied by manatees. The upper estimate for the number of manatees present in potentially impacted areas (within our study area) was estimated with our model to be 74 (95%CI 46 to 107). This upper estimate for the number of manatees was conditioned on the upper 95%CI value of the occupancy rate. In other words, based on our estimates, it is highly probable that there were 107 or fewer manatees in our study area during the time of our surveys. Because our analyses apply to habitats considered likely manatee habitats, our inference is restricted to these sites and to the time frame of our surveys. Given that manatees may be hard to see during aerial surveys, it was important to account for imperfect detection. The approach that we described can be useful for determining the best allocation of resources for monitoring and conservation.

Manatee vibrissae: evidence for a "lateral line" function.

Aquatic mammals use vibrissae to detect hydrodynamic stimuli over a range from 5 to 150 Hz, similar to the range detected by lateral line systems in fishes and amphibians. Manatees possess ∼5,300 vibrissae distributed over the body, innervated by ∼209,000 axons. This extensive innervation devoted to vibrissae follicles is reflected in enlarged, elaborate somatosensory regions of the gracile, cuneate, and Bischoff's brain-stem nuclei, ventrobasal thalamus, and presumptive somatosensory cortex. Our preliminary psychophysical testing indicates that in Florida and Antillean manatees the Weber fraction for detection thresholds for grating textures ranges from 0.025 to 0.14. At the lower end of this range, sensitivity is comparable to human index finger thresholds. For hydrodynamic stimuli of 5-150 Hz, detection threshold levels for manatees using facial or postfacial vibrissae were substantially lower than those reported for harbor seals and similar to reports of sensitivity for the lateral line systems of some fish. Our findings suggest that the facial and postfacial vibrissae are used to detect hydrodynamic stimuli, whereas only the facial vibrissae are used for direct contact investigation.

Auditory temporal resolution of a wild white-beaked dolphin (Lagenorhynchus albirostris).

Adequate temporal resolution is required across taxa to properly utilize amplitude modulated acoustic signals. Among mammals, odontocete marine mammals are considered to have relatively high temporal resolution, which is a selective advantage when processing fast traveling underwater sound. However, multiple methods used to estimate auditory temporal resolution have left comparisons among odontocetes and other mammals somewhat vague. Here we present the estimated auditory temporal resolution of an adult male white-beaked dolphin, (Lagenorhynchus albirostris), using auditory evoked potentials and click stimuli. Ours is the first of such studies performed on a wild dolphin in a capture-and-release scenario. The white-beaked dolphin followed rhythmic clicks up to a rate of approximately 1,125-1,250 Hz, after which the modulation rate transfer function (MRTF) cut-off steeply. However, 10% of the maximum response was still found at 1,450 Hz indicating high temporal resolution. The MRTF was similar in shape and bandwidth to that of other odontocetes. The estimated maximal temporal resolution of white-beaked dolphins and other odontocetes was approximately twice that of pinnipeds and manatees, and more than ten-times faster than humans and gerbils. The exceptionally high temporal resolution abilities of odontocetes are likely due primarily to echolocation capabilities that require rapid processing of acoustic cues.

Comments on "Intraspecific and geographic variation of West Indian manatee (Trichechus manatus spp.) vocalizations" [J. Acoust. Soc. Am. 114, 66-69 (2003)].

This letter concerns the paper "Intraspecific and geographic variation of West Indian manatee (Trichechus manatus spp.) vocalizations" [Nowacek et al., J. Acoust. Soc. Am. 114, 66-69 (2003)]. The purpose here is to correct the fundamental frequency range and information on intraindividual variation in the vocalizations of Amazonian manatees reported by Nowacek et al. (2003) in citing the paper "Signature information and individual recognition in the isolation calls of Amazonian manatees, Trichechus inunguis (Mammalia: Sirenia)" [Sousa-Lima et al., Anim. Behav. 63, 301-310 (2002)].

Temporal resolution of the Florida manatee (Trichechus manatus latirostris) auditory system.

Auditory evoked potential (AEP) measurements of two Florida manatees (Trichechus manatus latirostris) were measured in response to amplitude modulated tones. The AEP measurements showed weak responses to test stimuli from 4 kHz to 40 kHz. The manatee modulation rate transfer function (MRTF) is maximally sensitive to 150 and 600 Hz amplitude modulation (AM) rates. The 600 Hz AM rate is midway between the AM sensitivities of terrestrial mammals (chinchillas, gerbils, and humans) (80-150 Hz) and dolphins (1,000-1,200 Hz). Audiograms estimated from the input-output functions of the EPs greatly underestimate behavioral hearing thresholds measured in two other manatees. This underestimation is probably due to the electrodes being located several centimeters from the brain.

Cone visual pigments of aquatic mammals.

It has long been hypothesized that the visual systems of animals are evolutionarily adapted to their visual environment. The entrance many millions of years ago of mammals into the sea gave these new aquatic mammals completely novel visual surroundings with respect to light availability and predominant wavelengths. This study examines the cone opsins of marine mammals, hypothesizing, based on previous studies [Fasick et al. (1998) and Levenson & Dizon (2003)], that the deep-dwelling marine mammals would not have color vision because the pressure to maintain color vision in the dark monochromatic ocean environment has been relaxed. Short-wavelength-sensitive (SWS) and long-wavelength-sensitive (LWS) cone opsin genes from two orders (Cetacea and Sirenia) and an additional suborder (Pinnipedia) of aquatic mammals were amplified from genomic DNA (for SWS) and cDNA (for LWS) by PCR, cloned, and sequenced. All animals studied from the order Cetacea have SWS pseudogenes, whereas a representative from the order Sirenia has an intact SWS gene, for which the corresponding mRNA was found in the retina. One of the pinnipeds studied (harp seal) has an SWS pseudogene, while another species (harbor seal) appeared to have an intact SWS gene. However, no SWS cone opsin mRNA was found in the harbor seal retina, suggesting a promoter or splice site mutation preventing transcription of the gene. The LWS opsins from the different species were expressed in mammalian cells and reconstituted with the 11-cis-retinal chromophore in order to determine maximal absorption wavelengths (lambda(max)) for each. The deeper dwelling Cetacean species had blue shifted lambda(max) values compared to shallower-dwelling aquatic species. Taken together, these findings support the hypothesis that in the monochromatic oceanic habitat, the pressure to maintain color vision has been relaxed and mutations are retained in the SWS genes, resulting in pseudogenes. Additionally, LWS opsins are retained in the retina and, in deeper-dwelling animals, are blue shifted in lambda(max).

Diaphragm structure and function in the Florida manatee (Trichechus manatus latirostris).

Relative to many other mammals, little is known about the functional morphology of the four extant species of the order Sirenia. In this study, 166 Florida manatee (Trichechus manatus latirostris) carcasses fresh enough to collect detailed anatomical information were examined to describe the gross anatomy of the diaphragm. Our results show that the Florida manatee's diaphragm differs from those of other mammals in that it: lies in a dorsal plane, rather than in the more typical transverse plane; is located dorsal to the heart and does not attach to the sternum; and attaches medially at the "I"-shaped central tendon to bony projections extending ventrally from the vertebral bodies, forming two distinct hemidiaphragms. The manatee's transverse septum is a separate structure that lies at a right angle to the diaphragm and separates the heart from the liver and other viscera. The extreme muscularity of the diaphragm and the ability of manatees to adjust their position in the water column with minimal external movement suggest that diaphragmatic contractions may change the volume of each pleural cavity to affect buoyancy, roll, and pitch. We also hypothesize that such contractions, in concert with contractions of powerful abdominal muscles, may compress gas in the massive large intestine, and thereby also contribute to buoyancy control.