Contribution of host species and pathogen clade to snake fungal disease hotspots in Europe.

Infectious diseases are influenced by interactions between host and pathogen, and the number of infected hosts is rarely homogenous across the landscape. Areas with elevated pathogen prevalence can maintain a high force of infection and may indicate areas with disease impacts on host populations. However, isolating the ecological processes that result in increases in infection prevalence and intensity remains a challenge. Here we elucidate the contribution of pathogen clade and host species in disease hotspots caused by Ophidiomyces ophidiicola, the pathogen responsible for snake fungal disease, in 21 species of snakes infected with multiple pathogen strains across 10 countries in Europe. We found isolated areas of disease hotspots in a landscape where infections were otherwise low. O. ophidiicola clade had important effects on transmission, and areas with multiple pathogen clades had higher host infection prevalence. Snake species further influenced infection, with most positive detections coming from species within the Natrix genus. Our results suggest that both host and pathogen identity are essential components contributing to increased pathogen prevalence.

Paranannizziopsis spp. Infection in Wild Vipers, Europe.

We describe the detection of Paranannizziopsis sp. fungus in a wild population of vipers in Europe. Fungal infections were severe, and 1 animal likely died from infection. Surveillance efforts are needed to better understand the threat of this pathogen to snake conservation.

Yolk removal generates hatching asynchrony in snake eggs.

Hatching synchrony is wide-spread amongst egg-laying species and is thought to enhance offspring survival, notably by diluting predation risks. Turtle and snake eggs were shown to achieve synchronous hatching by altering development rates (where less advanced eggs may accelerate development) or by hatching prematurely (where underdeveloped embryos hatch concurrently with full-term embryos). In Natricine snakes, smaller eggs tend to slow down metabolism throughout incubation in order to hatch synchronously with larger eggs. To explore the underlying mechanism of this phenomenon we experimentally manipulated six clutches, where half of the eggs were reduced in mass by removing 7.2% of yolk, and half were used as the control. The former experienced higher heart rates throughout the incubation period, hatched earlier and produced smaller hatchlings than the latter. This study supports the idea that developmental rates are related to egg mass in snake eggs and demonstrates that the relationship can be influenced by removing yolk after egg-laying. The shift in heart rates however occurred in the opposite direction to expected, with higher heart rates in yolk-removed eggs resulting in earlier hatching rather than lower heart rates resulting in synchronous hatching, warranting further research on the topic.

Only child syndrome in snakes: Eggs incubated alone produce asocial individuals.

Egg-clustering and communal nesting behaviours provide advantages to offspring. Advantages range from anti-predatory benefits, maintenance of moisture and temperature levels within the nest, preventing the eggs from rolling, to enabling hatching synchrony through embryo communication. It was recently suggested that embryo communication may extend beyond development fine-tuning, and potentially convey information about the quality of the natal environment as well as provide an indication of forthcoming competition amongst siblings, conspecifics or even heterospecifics. Here we show that preventing embryos from communicating not only altered development rates but also strongly influenced post-natal social behaviour in snakes. Clutches of water snakes, Natrix maura, were split evenly into half-clutches and incubated as (1) clusters (i.e. eggs in physical contact with each other) or (2) as single eggs placed in individual goblets (i.e. no physical contact amongst sibling eggs). Single incubated eggs produced less-sociable young snakes than their siblings that were incubated in a cluster: the former were more active, less aggregated and physically contacted each other less often than the latter. Potential long-term effects and evolutionary drivers for this new example of informed dispersal are discussed.

Heartbeat, embryo communication and hatching synchrony in snake eggs.

Communication is central to life at all levels of complexity, from cells to organs, through to organisms and communities. Turtle eggs were recently shown to communicate with each other in order to synchronise their development and generate beneficial hatching synchrony. Yet the mechanism underlying embryo to embryo communication remains unknown. Here we show that within a clutch, developing snake embryos use heart beats emanating from neighbouring eggs as a clue for their metabolic level, in order to synchronise development and ultimately hatching. Eggs of the water snake Natrix maura increased heart rates and hatched earlier than control eggs in response to being incubated in physical contact with more advanced eggs. The former produced shorter and slower swimming young than their control siblings. Our results suggest potential fitness consequences of embryo to embryo communication and describe a novel driver for the evolution of egg-clustering behaviour in animals.

Myth busting? Effects of embryo positioning and egg turning on hatching success in the water snake Natrix maura.

It is a common belief that reptile eggs should not be turned after oviposition once the embryo has attached itself to the inner membrane of the shell as it might kill developing embryos. Here, we used 338 eggs from 32 clutches of the water snake Natrix maura to (1) thoroughly describe natural clutch arrangement, (2) experimentally assess the effects of natural embryo positioning and (3) egg turning on embryo metabolism, hatching success, and hatchling phenotype. Clutches contained, on average, 59% of embryos located at the top, 28% at the bottom, and 14% on a side of the egg. Larger females laid larger clutches with higher proportion of top located embryos. Top embryos displayed higher metabolic rates (heart rates), shorter incubation time, and produced lighter and shorter snakes than bottom embryos. Egg turning did not significantly influence egg development, hatching success or hatchling phenotypes. However, post-birth mortality was significantly higher in turned (37.5%) compared to unturned (4.5%) embryos, providing support to the common belief that eggs should not be moved from their natural position.

Ultrasound imaging of the inguinal region of adult male loggerhead sea turtles (Caretta caretta).

The biology and reproductive anatomy of male loggerhead sea turtles (Caretta caretta) have been difficult to study. The principle method for evaluation of the coelomic cavity in both adult and juvenile male sea turtles is celioscopy. The purpose of this study was to describe the technique and structures seen when scanning the inguinal region of live, wild-caught, adult male loggerhead sea turtles and to compare these findings to those resulting from celioscopy and necropsy. Twenty-one adult male loggerhead sea turtles were collected by trawling in the Cape Canaveral shipping channel in April 2007. All turtles were placed in dorsal recumbency and imaged with a Sonosite 180 Vet Plus (Sonosite, Inc., Bothell, Washington 98021, USA) and a microconvex, 4-7-MHz curvilinear array probe. The inguinal region was divided into four quadrants: cranial, lateral, medial, and caudal. Celioscopy was performed on 13 turtles, and biopsies were obtained of the testes and the epididymides to confirm correct identification of the structures. In the cranial aspect of the inguinal region, the urinary bladder and large and small intestines were identified. In the lateral inguinal region, the lung and kidney were seen. In the medial aspect of the inguinal region, the testis and epididymis were routinely identified. In the caudal aspect of the inguinal region, the coxofemoral joint was seen. A small learning curve was required; however, correlation with celioscopy and biopsy showed that consistent, repeatable identification of caudal coelomic structures was easily achieved. Ultrasound provided an inexpensive, rapid, noninvasive method to evaluate the reproductive anatomy of live-captured, male loggerhead sea turtles.

Diamondback terrapins, Malaclemys terrapin, as a sentinel species for monitoring mercury pollution of estuarine systems in South Carolina and Georgia, USA.

Total mercury concentrations were measured in diamondback terrapin blood and scutes collected from four sites in South Carolina, USA, and at a superfund site in Brunswick, Georgia, USA. There was a strong correlation between mercury concentrations in the two terrapin body compartments (Kendall's tau = 0.79, p < 0.001). Mercury concentrations in terrapin scute and blood and in salt marsh periwinkles, Littoraria irrorata, were significantly higher in Brunswick (scute x = 3810.2 ng/g, blood x = 746.2 ng/g) than from all other sites (scute x = 309.5 ng/g, blood x = 43.2 ng/g, p < 0.001). Seasonal fluctuations of total mercury in the blood and scutes of terrapins collected in the Ashley River, South Carolina, were significantly lower in August than in April, June, or October in blood (p < 0.001); however, scute concentrations did not vary seasonally. Overall, we found higher concentrations of mercury in the scutes of females than males (n = 32, p < 0.05). Larger females may preferentially prey on larger food items, like large periwinkles, which had significantly higher mercury levels in their body tissues than smaller periwinkles (p < 0.001). Methylmercury levels in terrapin scutes were measured, revealing that 90% of the total mercury stored in this compartment was in the organic form. A methylmercury biomagnification factor of 173.5 was calculated from snails to terrapin scutes, and we found that mercury levels in scutes were representative of the mercury levels in other compartments of the ecosystem. These findings show that terrapin scutes are good predictors of mercury pollution and that this species could be used as a bioindicator for assessing mercury contamination of estuarine systems.

Putative involvement of crustacean hyperglycemic hormone isoforms in the neuroendocrine mediation of osmoregulation in the crayfish Astacus leptodactylus.

This study investigates the involvement of eyestalk neuroendocrine factors on osmoregulation in the crayfish Astacus leptodactylus maintained in freshwater. Eyestalk removal was followed by a significant decrease in hemolymph osmolality and Na(+) concentration and by a 50% increase in mass after one molting cycle. Several neurohormones have been isolated from the sinus gland through high-performance liquid chromatography (HPLC), and different crustacean hyperglycemic hormone (CHH)-related peptides, including stereoisomers (L-CHH and D-Phe(3) CHH), have been identified by direct enzyme-linked immunosorbent assay (ELISA). A glucose quantification bioassay demonstrated a strong hyperglycemic activity following injection of the immunoreactive chromatographic fractions and showed that the D-Phe(3) CHH was the most efficient. Destalked crayfish were then injected with purified CHH HPLC fractions. The D-Phe(3) CHH fraction significantly increased the hemolymph osmolality and Na(+) content 24 h after injection. Two other CHH-related peptides caused a smaller increase in Na(+) concentration. No significant variation was observed in hemolymph Cl(-) concentration following injection of any of the CHH isoforms. These results constitute the first observation of the effects of a CHH isoform, specifically the D-Phe(3) CHH, on osmoregulatory parameters in a freshwater crustacean. The effects of eyestalk ablation and CHH injection on osmoregulation and the identification of different CHH-related peptides and isoforms in crustaceans are discussed.