Characterization of seasonal reproductive and stress steroid hormones in wild Radiated Tortoises, Astrochelys radiata.

The critically endangered Radiated Tortoise (Astrochelys radiata) is endemic to the southern coastlines of Madagascar. Once common, wild populations of this tortoise have undergone dramatic declines in recent years. Although there have been studies documenting reproductive activities, reproductive physiological parameters are unknown yet may be crucial in the recovery of the species. Over four research seasons in remote field locations native to A. radiata, we surveyed for, radio-tracked, and sampled wild, free ranging tortoises. We sampled and measured stress and reproductive parameters (corticosterone [CORT], testosterone [T], estradiol-17ß [E2], and progesterone [P]) in 311 plasma samples from 203 wild A. radiata, capturing their active period. Generally, hormone concentrations were associated with body condition, temperature, and humidity. There was wide variation in CORT that varied monthly and by group. Juvenile tortoises maintained more than twice the mean basal CORT concentrations than either adult sex, with the most dramatic distinctions in the middle of the wet season. For adult sex hormones, the last months of the dry season and into the wet season when ground humidities are low and just begin to rise prior to temperature declines, male T concentrations gradually increased to a peak before returning to near undetectable values into the dry season. We had limited data for T concentrations in females, but found average T concentrations were much lower than in males and positively correlated with larger female home range sizes. For female hormone cycles, E2 also peaked in the early 1/3 of the wet season along with male T, and was followed by an uptick in P which correlates to the putative ovulatory cycle. Females tracked over four years showed variation in patterns of P, indicating that number and frequency of clutches vary. Our results suggest that 1) there is high species plasticity in response to stress; 2) A. radiata reproductive cycling is somewhat dissociated with courtship timing and is instead triggered by environmental cues; and 3) individual female reproductive output is irregular. This study is oone of the first to document and describe multi-year seasonal stress and reproductive hormones in a free-ranging Malagasy chelonian. These data may be used to identify key high-production habitats for conservation, and aide in captive management and reproduction in assurance colonies for species health and survival.

Comparative ecophysiology of a critically endangered (CR) ectotherm: Implications for conservation management.

Captive breeding is a vital conservation tool for many endangered species programs. It is often a last resort when wild animal population numbers drop to below critical minimums for natural reproduction. However, critical ecophysiological information of wild counterparts may not be well documented or understood, leading to years of minimal breeding successes. We collected endocrine and associated ecological data on a critically endangered ectotherm concurrently in the wild and in captivity over several years. We tracked plasma concentrations of steroid stress and reproductive hormones, body condition, activity, and environmental parameters in three populations (one wild and two geographically distinct captive) of ploughshare tortoise (Astrochelys yniphora). Hormone profiles along with environmental and behavioral data are presented and compared. We show that animals have particular seasonal environmental requirements that can affect annual reproduction, captivity affects reproductive state, and sociality may be required at certain times of the year for breeding to be successful. Our data suggest that changes in climatic conditions experienced by individuals, either due to decades-long shifts or hemispheric differences when translocated from their native range, can stifle breeding success for several years while the animals physiologically acclimatize. We also found that captivity affects stress (plasma corticosterone) and body condition of adults and juveniles differently and seasonally. Our results indicate that phenotypic plasticity in reproduction and behavior is related to environmental cues in long-lived ectotherms, and detailed ecophysiological data should be used when establishing and improving captive husbandry conditions for conservation breeding programs. Further, considering the recent revelation of this tortoises' possible extirpation from the wild, these data are critically opportune and may be key to the survival of this species.

Stress response to handling is short lived but may reflect personalities in a wild, Critically Endangered tortoise species.

We investigated the acute stress response associated with animal personalities by measuring plasma glucocorticoids throughout handling and collected ~2 years of movement and behavioural data in a wild, Critically Endangered animal, Astrochelys radiata (radiated tortoise). To determine whether our standard, brief conscientious handling procedures induce a stress response in our target species, we applied a stressor by way of initial animal processing and deployment of telemetry equipment. During surveys and processing, we sampled animals immediately upon detection, again after completing transmitter attachment and processing, and a final time the following day. We then used radiotelemetry to follow a subset of the animals for 22 months while collecting behavioural, climatic and location data. We found that brief and conscientious handling did not illicit consistent changes in plasma concentrations of the stress hormone corticosterone (CORT) but did reveal tremendous individual variation in response. The CORT concentration ranged more than 200-fold after imposing the stressor and returned to near-baseline values by the following day. When we accounted for the wide variation by calculating the degree of each individual's stress response relative to its baseline over its processing time, we discovered two non-overlapping physiological response types; those in which CORT concentrations increased dramatically in response to handling (219 ± 89.8 pg/ml/min) and those in which CORT varied only slightly (5.3 ± 8.9 pg/ml/min). The response types (strong vs. mild) also predicted body condition, home range size, activity, and behavioural tendencies. The degree of the individual's stress response in this species may be one component of correlated physiological and behavioural traits (animal personalities), which have previously been obscured in other chelonian studies by the use of mean values and should be considered in future conservation management applications for chelonian species.

Radio Transmitter Implantation and Movement in the Wild Timber Rattlesnake ( Crotalus horridus ).

Radiotelemetry transmitters have become critical to studies of wildlife ecology. However, little is known about how transmitter implantation surgery affects the mobility of some species, including the timber rattlesnake, Crotalus horridus . Tracking snake movement can provide insights into the effects of transmitter implantation. During 2007-11, 71 radio transmitters were surgically implanted intracoelomically in 47 timber rattlesnakes. Over 20 of these snakes underwent surgery at least twice in 5 yr to replace old transmitters. Surgeries were performed under general anesthesia with a local nerve block at the site of implantation, 20 cm cranial to the cloaca. Snakes were also administered postsurgical meloxicam and enrofloxacin every 24 h for three doses. Two to five days after surgery, snakes were released at their original locations and radiotracked regularly during the active seasons (April-October 2007-11). Average daily movement data (distance traveled) were compiled for each snake. Snakes undergoing transmitter surgery in a given year did not differ significantly in distance traveled compared to snakes that had transmitters but did not have surgery in that year. Distance traveled for each snake did not differ before or after surgery or between weeks 1 and 2 postsurgery, indicating that the transmitter implantation did not alter snake movement.