Fine-scale hormonal patterns associated with birth and maternal care in the cottonmouth (Agkistrodon piscivorus), a North American pitviper snake.

Steroid hormones regulate many aspects of reproductive physiology and behavior, including parental care. Reptiles display a variety of egg- and neonate-directed parental behaviors, yet few studies have addressed their endocrine correlates. Viviparous female pitvipers remain at the birth site with their young for one to two weeks until neonates complete their first shed cycle ('ecdysis'). To study possible relationships between steroid hormones and these behaviors, we conducted a captive study on wild-caught pregnant cottonmouths. Females were divided into two treatment groups: Maternal Attendance (MA) - females were allowed a maternal attendance period, where neonates were left with the mother until they completed ecdysis and then were removed; Separated (SE) - females had their neonates removed within 24h of birth. Serial blood samples were collected from MA females at various points during and after attendance; SE females had samples collected on a similar temporal schedule. Plasma levels of progesterone (P), estradiol (E2), testosterone (T), and corticosterone (CORT) were measured in all samples. We did not find a difference in the overall pattern of P, E2, or T between MA and SE females; however, MA females exhibited a significant peak in CORT on the day that neonates shed that was not observed in SE females. It is possible that the elevated CORT observed in MA females was stimulated by increased activity and/or changing chemical cues of shedding neonates. Based on evidence that free-ranging pitvipers cease MA when all offspring complete ecdysis, we hypothesize that CORT has a role in signaling mothers to terminate care and disperse.

Facultative parthenogenesis discovered in wild vertebrates.

Facultative parthenogenesis (FP)-asexual reproduction by bisexual species-has been documented in a variety of multi-cellular organisms but only recently in snakes, varanid lizards, birds and sharks. Unlike the approximately 80 taxa of unisexual reptiles, amphibians and fishes that exist in nature, FP has yet to be documented in the wild. Based on captive documentation, it appears that FP is widespread in squamate reptiles (snakes, lizards and amphisbaenians), and its occurrence in nature seems inevitable, yet the task of detecting FP in wild individuals has been deemed formidable. Here we show, using microsatellite DNA genotyping and litter characteristics, the first cases of FP in wild-collected pregnant females and their offspring of two closely related species of North American pitviper snakes-the copperhead (Agkistrodon contortrix) and cottonmouth (Agkistrodon piscivorus). Our findings support the view that non-hybrid origins of parthenogenesis, such as FP, are more common in squamates than previously thought. With this confirmation, FP can no longer be viewed as a rare curiosity outside the mainstream of vertebrate evolution. Future research on FP in squamate reptiles related to proximate control of induction, reproductive competence of parthenogens and population genetics modelling is warranted.

Reproduction in female copperhead snakes (Agkistrodon contortrix): plasma steroid profiles during gestation and post-birth periods.

We investigated levels of plasma progesterone (P4), 17ß-estradiol (E2), testosterone (T), and corticosterone (CORT) during gestation and post-birth periods in wild-collected female copperhead snakes (Viperidae; Agkistrodon contortrix). We also sought to determine whether CORT levels at (or near) birth dramatically increase and were correlated with duration of labor and litter size. Specifically, pregnant subjects (N = 14) were collected during early- to mid-gestation, held in the laboratory, and repeatedly bled to obtain plasma for steroid analyses. Progesterone showed significant changes during gestation, with the highest levels at the onset of sampling (circa 50 days prior to birth); P4 progressively declined up to parturition, and basal levels were observed thereafter. At the onset of sampling, E2 was at peak levels and fell sharply at circa 30 days prior to birth, a trend observed throughout the post-birth sampling period. Throughout the entire sampling period, T was undetectable. Although CORT showed no significant changes during gestation and several days following parturition, there was a highly significant peak at the time of birth. Our findings mirror the results of previous studies on pregnancy and steroid hormones of other live-bearing snakes, lizards, and mammals. As expected, there was a significant relationship between duration of labor and litter size; however, although levels of CORT did not achieve significance, there was a positive trend with litter size. We suggest that elevation of CORT at birth is involved in the mobilization and regulation of energy stores necessary for the physiological process of parturition and as a possible mechanism to trigger birth.

The reproductive biology of male cottonmouths (Agkistrodon piscivorus): do plasma steroid hormones predict the mating season?

To better understand the proximate causation of the two major types of mating seasons described for North American pitvipers, we conducted a field study of the cottonmouth (Agkistrodon piscivorus) in Georgia from September 2003 to May 2005 that included an extensive observational regime and collection of tissues for behavioral, anatomical, histological, and hormone analysis. Enzyme immunoassays (EIA) of plasma samples and standard histological procedures were conducted on reproductive tissues. Evidence from the annual testosterone (T) and sexual segment of the kidney (SSK) cycle and their relationship to the spermatogenic cycle provide correlative evidence of a unimodal mating pattern in this species of pitviper, as these variables consistently predict the mating season in all snake species previously examined under natural conditions. In most reptiles studied to date, high plasma levels of T and corticosterone (CORT) coincide during the mating period, making the cottonmouth an exception to this trend; we suggest two possible explanations for increased CORT during spring (regulation of a spring basking period), and decreased CORT during summer (avoiding reproductive behavioral inhibition), in this species.