Skin glands of an aquatic salamander vary in size and distribution and release antimicrobial secretions effective against chytrid fungal pathogens.

Amphibian skin is unique among vertebrate classes, containing a large number of multicellular exocrine glands that vary among species and have diverse functions. The secretions of skin glands contain a rich array of bioactive compounds including antimicrobial peptides (AMPs). Such compounds are important for amphibian innate immune responses and may protect some species from chytridiomycosis, a lethal skin disease caused by the fungal pathogens Batrachochytrium dendrobatidis (Bd) and Batrachochytrium salamandrivorans (Bsal). While the bioactivity of skin secretions against Bd has been assessed for many amphibian taxa, similar studies are lacking for Bsal, a chytrid fungus that is especially pathogenic for salamanders. We studied the skin glands and their potential functions in an aquatic salamander, the three-toed amphiuma (Amphiuma tridactylum). Skin secretions of captive adult salamanders were analyzed by RP-HPLC and tested against the growth of Bd and Bsal using in vitro assays. We found that compounds within collected skin secretions were similar between male and female salamanders and inhibited the growth of Bd and Bsal. Thus, skin secretions that protect against Bd may also provide protection against Bsal. Histological examination of the skin glands of preserved salamanders revealed the presence of enlarged granular glands concentrated within caudal body regions. A site of potential gland specialization was identified at the tail base and may indicate specialized granular glands related to courtship and communication.

The influence of thermal biology on road mortality risk in snakes.

Road mortality is a significant threat to terrestrial vertebrates in many areas, and the novel thermal environment of black-topped roads may represent ecological traps for some species and demographic groups. We investigated the relationship between ambient temperature and on-road detection in a snake assemblage in southeastern Louisiana by comparing observations of live snakes on a black-topped road, across measurements of air temperature and road temperature on survey days. Analyses indicated on-road detection of snakes was significantly influenced by ambient temperature conditions for five snake species. Additionally, road temperatures, and the difference between air and road temperatures, were strong drivers of on-road snake detections. Permutation analysis methods revealed that significant temperature related group (species or sex) structure exists in occurrences of snakes on the roadway, and that road temperature was the strongest driver of species differences. We also compared how air and road temperatures affected occurrence on the road between sexes in the colubrid snakes Nerodia fasciata, Nerodia cyclopion, Thamnophis proximus, and Pantherophis obsoletus. Males and females of the viviparous species N. fasciata, N. cyclopion, and T. proximus diverged significantly in temperature preferences, with females found under warmer conditions, while males and females of the oviparous species P. obsoletus did not. Road temperature was also the strongest driver of differences between sexes. Our results indicate that black-topped roads are an ecological trap that is heavily influenced by sex, reproductive condition, and species specific thermoregulatory requirements, particularly for viviparous species.

Serum complement activity in the three-toed amphiuma (Amphiuma tridactylum).

Some animals routinely endure serious injuries from predators or during intraspecific territorial conflicts. Such is the case for Amphiuma tridactylum, an aquatic salamander that lives in an environment rich in potentially infectious microbes, apparently with rare or no pathogenic infection. Some vertebrates possess innate immune mechanisms, but whether this is the case for Amphiuma is unknown. To assess this potential, plasma from 19 A. tridactylum was pooled and used for characterisation of serum complement activity. The ability of A. tridactylum plasma to hemolyse unsensitised sheep red blood cells (SRBCs) was titer-dependent, with low activity observed even at high plasma titers. The kinetic characterisation of SRBC hemolysis revealed that significant activity could be measured within 10min of incubation, and maximal activity occurred within 60min. The SRBC hemolysis by A. tridactylum plasma was also temperature-dependent, with maximal activity at 30°C. In addition, this activity was sensitive to mild heat treatment, with 96% of activity inhibited by incubation at 56°C for 30min. The SRBC hemolysis could also be inactivated by pretreatment of the plasma with proteases, indicating that this activity was protein dependent. The activity required divalent metals ions, with activity inhibited by EDTA, citrate, or phosphate. However, the chelator-inhibited activity could be restored by the addition of excess Ca(2+) or Mg(2+), but not Cu(2+) or Ba(2+), indicating specificity of the divalent metal ion requirement. The sensitivity to heat, proteases, and divalent metal ion chelators strongly suggests that A. tridactylum plasma-mediated hemolysis of SRBCs is mediated by the serum complement system of proteins.

Variation in pupil diameter in North American Gartersnakes (Thamnophis) is regulated by immersion in water, not by light intensity.

A variable pupil generally regulates the amount of incoming light available for image formation on the retina. However, some of the semi-aquatic snakes (North American Gartersnakes, Thamnophis) that forage in relatively low light conditions reduce the pupil aperture in response to submergence underwater at the expense incoming light. Given that these snakes have all-cone retinas, reduction of incoming light because of pupillary constriction upon immersion seems counterintuitive. To test the effect of light and water on pupil aperture, three species of North American Gartersnakes (T. atratus, T. hammondii, and T. sirtalis) were exposed to nine light intensities in air and water. There was no effect of light on relative pupil aperture for any species. However, all three species showed a significant reduction in pupil aperture upon submergence underwater. The lack of a light response is surprising, and may be related to the method of accommodation in snakes. Snakes lack a ciliary muscle, and move the lens by constricting the pupil, which increases pressure in the posterior chamber and pushes the lens forward. Upon submergence, the snakes may be attempting to overcome the change in refractive index and defocus imposed by the water, by constricting the pupil. Thus, having the iris muscle involved in accommodation may preclude it from much of a light regulating function.