Two heads are not always better than one: Craniofacial and axial bifurcation in cheloniid embryos and hatchlings (Chelonia mydas and Caretta caretta).

Morphological data on craniofacial and axial bifurcation in sea turtles is not well documented in the literature. Here, we use micro-computed tomography (µ-CT) imaging to describe the body, skull, and vertebral morphology in axially-bifurcated cheloniid sea turtle embryos and hatchlings (Chelonia mydas and Caretta caretta) from south Florida beaches. We describe three types of craniofacial and axial bifurcations: bifacial, bicephalic, and bicephalic with biaxial duplication ranging from facial bones to the sacrum. We predicted smaller body dimensions in bifacial and bicephalic embryos and hatchlings compared with their normal counterparts. In addition, we hypothesized that bicephalic individuals would have greater rostral deviation angles than bifacial animals, and that vertebral dimensions would vary between the control and anomalous embryos and hatchlings. Among hatchlings (developmental Stage 31), we found that maximum curved carapace length and curved carapace width were greatest in the control specimens when compared with the anomalous specimens. Overall, we found that rostral deviations were smaller in bifacial animals compared with their bicephalic counterparts. Right and left rostral deviations were symmetrical or nearly symmetrical in all bifacial and bicephalic specimens. Among C. caretta, we found that bicephalic animals had greater standardized vertebral measurements than their bifacial conspecifics. In bifacial animals, bifurcation extended to either the frontal or parietal skull bones, while duplication extended to C5 vertebrae and T8 vertebrae in bicephalic animals. This study provides an in-depth description of anatomical alterations associated with these abnormalities. Prognosis of these organisms is poor; however, understanding the prevalence of these malformations can allow for better assessments of population health, as numerous environmental factors are known to lead to these changes.

Damsels in Distress: Oil Exposure Modifies Behavior and Olfaction in Bicolor Damselfish (Stegastes partitus).

In fishes, olfactory cues evoke behavioral responses that are crucial to survival; however, the receptors, olfactory sensory neurons, are directly exposed to the environment and are susceptible to damage from aquatic contaminants. In 2010, 4.9 million barrels of crude oil were released into the northern Gulf of Mexico from the Deepwater Horizon disaster, exposing marine organisms to this environmental contaminant. We examined the ability of bicolor damselfish (Stegastes partitus), exposed to the water accommodated fraction (WAF) of crude oil, to respond to chemical alarm cue (CAC) using a two-channel flume. Control bicolor damselfish avoided CAC in the flume choice test, whereas WAF-exposed conspecifics did not. This lack of avoidance persisted following 8 days of control water conditions. We then examined the physiological response to CAC, brine shrimp rinse, bile salt, and amino acid cues using the electro-olfactogram (EOG) technique and found that WAF-exposed bicolor damselfish were less likely to detect CAC as an olfactory cue but showed no difference in EOG amplitude or duration compared to controls. These data indicate that a sublethal WAF exposure directly modifies detection and avoidance of CAC beyond the exposure period and may suggest reduced predator avoidance behavior in oil-exposed fish in the wild.

Leptin's effect on taste bud calcium responses and transmitter secretion.

Leptin, a peptide hormone released by adipose tissue, acts on the hypothalamus to control cravings and appetite. Leptin also acts to decrease taste responses to sweet substances, though there is little detailed information regarding where leptin acts in the taste transduction cascade. The present study examined the effects of leptin on sweet-evoked responses and neuro transmitter release from isolated taste buds. Our results indicate that leptin moderately decreased sweet-evoked calcium mobilization in isolated mouse taste buds. We also employed Chinese hamster ovary biosensor cells to examine taste transmitter release from isolated taste buds. Leptin reduced ATP and increased serotonin release in response to sweet stimulation. However, leptin has no effect on bitter-evoked transmitter release, further showing that the action of leptin is sweet specific. Our results support those of previous studies, which state that leptin acts on taste tissue via the leptin receptor, most likely on Type II (Receptor) cells, but also possibly on Type III (Presynaptic) cells.

The somatotopic organization of the olfactory bulb in elasmobranchs.

The olfactory bulbs (OBs) are bilaterally paired structures in the vertebrate forebrain that receive and process odor information from the olfactory receptor neurons (ORNs) in the periphery. Virtually all vertebrate OBs are arranged chemotopically, with different regions of the OB processing different types of odorants. However, there is some evidence that elasmobranch fishes (sharks, rays, and skates) may possess a gross somatotopic organization instead. To test this hypothesis, we used histological staining and retrograde tracing techniques to examine the morphology and organization of ORN projections from the olfactory epithelium (OE) to the OB in three elasmobranch species with varying OB morphologies. In all three species, glomeruli in the OB received projections from ORNs located on only the three to five lamellae situated immediately anterior within the OE. These results support that the gross arrangement of the elasmobranch OB is somatotopic, an organization unique among fishes and most other vertebrates. In addition, certain elasmobranch species possess a unique OB morphology in which each OB is physically subdivided into two or more "hemi-olfactory bulbs." Somatotopy could provide a preadaptation which facilitated the evolution of olfactory hemibulbs in these species.

Sensitivity and specificity of the olfactory epithelia of two elasmobranch species to bile salts.

Odor detection in vertebrates occurs when odorants enter the nose and bind to molecular olfactory receptors on the cilia or microvilli of olfactory receptor neurons (ORNs). Several vertebrate groups possess multiple, morphologically distinct types of ORNs. In teleost fishes, these different ORN types detect specific classes of biologically relevant odorants, such as amino acids, nucleotides and bile salts. For example, bile salts are reported to be detected exclusively by ciliated ORNs. The olfactory epithelium of elasmobranch fishes (sharks, rays and skates) is comprised of microvillous and crypt ORNs, but lacks ciliated ORNs; thus, it was questioned whether the olfactory system of this group of fishes is capable of detecting bile salts. The present investigation clearly indicates that the olfactory system of representative shark and stingray species does detect and respond to bile salts. Additionally, these species detect glycine-conjugated, taurine-conjugated and non-conjugated bile salts, as do teleosts. These elasmobranchs are less sensitive to the tested bile salts than reported for both agnathans and teleosts, but this may be due to the particular bile salts selected in this study, as elasmobranch-produced bile salts are commercially unavailable. Cross-adaptation experiments indicate further that the responses to bile salts are independent of those to amino acids, a major class of odorant molecules for all tested fishes.

Olfactory morphology and physiology of elasmobranchs.

Elasmobranch fishes are thought to possess greater olfactory sensitivities than teleost fishes due in part to the large amount of epithelial surface area that comprises their olfactory organs; however, direct evidence correlating the size of the olfactory organ to olfactory sensitivity is lacking. This study examined the olfactory morphology and physiology of five distantly related elasmobranch species. Specifically, we quantified the number of lamellae and lamellar surface area (as if it were a flat sheet, not considering secondary lamellae) that comprise their olfactory organs. We also calculated the olfactory thresholds and relative effectiveness of amino acid odorants for each species. The olfactory organs varied in both the number of lamellae and lamellar surface area, which may be related to their general habitat, but neither correlated with olfactory threshold. Thresholds to amino acid odorants, major olfactory stimuli of all fishes, ranged from 10⁻9·° to 10⁻6·9 mol l⁻¹, which indicates that these elasmobranch species demonstrate comparable thresholds with teleosts. In addition, the relative effectiveness of amino acid stimuli to the olfactory organ of elasmobranchs is similar to that previously described in teleosts with neutral amino acids eliciting significantly greater responses than others. Collectively, these results indicate parallels in olfactory physiology between these two groups of fishes.