title: A hormone priming regimen and hibernation affect oviposition in the boreal toad (Anaxyrus boreas boreas).
abstract: Declines of the southern Rocky Mountain population of boreal toad (Anaxyrus boreas boreas) have led to the establishment of a captive assurance population and reintroduction program, in an attempt to preserve and propagate this geographically isolated population. One of the unique adaptations of this species is its ability to survive in cold environments by undergoing long periods of hibernation. In captivity, hibernation can be avoided altogether, decreasing morbidity caused by compromised immune systems. However, it is not entirely clear how essential hibernation is to reproductive success. In this study, the effects of hibernation versus nonhibernation, and exogenous hormones on oviposition, were examined in boreal toad females in the absence of males. In the summers of 2011 and 2012, 20 females housed at Mississippi State University were treated with a double priming dose of hCG and various ovulatory doses of hCG and LH-releasing hormone analog but denied hibernation. Exogenous hormones, in the absence of hibernation, could not induce oviposition over two breeding seasons (2011-2012). In contrast, during the summer of 2012 and 2013, 17 of 22 females (77%) housed at the Native Aquatic Species Restoration Facility (Alamosa, CO, USA) oviposited after they were treated with two priming doses of hCG (3.7 IU/g each) and a single ovulation dose of hCG (13.5 IU/g) and LH-releasing hormone analog (0.4 μg/g) after hibernation. There was a significant difference in oviposition between females that were hibernated and received hormones (2012, P < 0.05 and 2013, P < 0.01) compared to hibernated control females. In 2013, 12 of 16 remaining Mississippi State University females from the same group used in 2011 and 2012 were hibernated for 1, 3, and 6 months, respectively and then treated with the same hormone regimen administered to females at the Native Aquatic Species Restoration Facility. Together, hibernation and hormone treatments significantly increased oviposition (P < 0.05), with 33% of females ovipositing. These results suggest that (1) hibernation is a key factor influencing oviposition that cannot be exclusively circumvented by exogenous hormones; (2) females do not require the presence of a male to oviposit after hormone treatments; and (3) longer hibernation periods are not beneficial for oviposition. The hormonal induction of oviposition in the absence of males and shorter hibernation periods could have important captive management implications for the boreal toad. Furthermore, the production of viable offspring by IVF where natural mating is limited could become an important tool for genetic management of this boreal toad captive population.
authors.affiliation: Department of Biochemistry, Molecular Biology, Entomology and Plant Pathology, Mississippi State University, Starkville, Mississippi, USA; Department of Conservation and Research, Memphis Zoo, Memphis, Tennessee, USA.
authors.forename: N E
authors.initials: NE
authors.lastname: Calatayud
authors.affiliation: Department of Biochemistry, Molecular Biology, Entomology and Plant Pathology, Mississippi State University, Starkville, Mississippi, USA; Department of Conservation and Research, Memphis Zoo, Memphis, Tennessee, USA.
authors.forename: C J
authors.initials: CJ
authors.lastname: Langhorne
authors.affiliation: Department of Biochemistry, Molecular Biology, Entomology and Plant Pathology, Mississippi State University, Starkville, Mississippi, USA.
authors.forename: A C
authors.initials: AC
authors.lastname: Mullen
authors.affiliation: Department of Biochemistry, Molecular Biology, Entomology and Plant Pathology, Mississippi State University, Starkville, Mississippi, USA; Department of Conservation and Research, Memphis Zoo, Memphis, Tennessee, USA.
authors.forename: C L
authors.initials: CL
authors.lastname: Williams
authors.affiliation: Colorado Department of Parks and Wildlife, Native Aquatic Species Restoration Facility, Alamosa, Colorado, USA.
authors.forename: T
authors.initials: T
authors.lastname: Smith
authors.affiliation: Department of Biochemistry, Molecular Biology, Entomology and Plant Pathology, Mississippi State University, Starkville, Mississippi, USA.
authors.forename: L
authors.initials: L
authors.lastname: Bullock
authors.affiliation: Department of Conservation and Research, Memphis Zoo, Memphis, Tennessee, USA.
authors.forename: A J
authors.initials: AJ
authors.lastname: Kouba
authors.affiliation: Department of Biochemistry, Molecular Biology, Entomology and Plant Pathology, Mississippi State University, Starkville, Mississippi, USA. Electronic address: swillard@cals.msstate.edu.
authors.forename: S T
authors.initials: ST
authors.lastname: Willard
mesh_terms: D000818:Animals; D002025:Bufonidae / Q000502:physiology*; D006063:Chorionic Gonadotropin / Q000494:pharmacology*; D005260:Female; D007987:Gonadotropin-Releasing Hormone / Q000494:pharmacology*; D006605:Hibernation / Q000502:physiology*; D010058:Oviposition / Q000187:drug effects* / Q000502:physiology; D013997:Time Factors
chemical_list: D006063:Chorionic Gonadotropin; D007987:Gonadotropin-Releasing Hormone
keywords: Hibernation; Hormone; IVF; LH-releasing hormone analog; Oviposition; hCG