Russian Collaborative Development of Reproduction Technologies for the Sustainable Management of Amphibian Biodiversity

Reproduction technologies(RTs)can provide for the reliable reproduction of amphibians,as well as perpetuation of species genetic variation with the use of biobanks.In 1982,in anticipation of the biodiversity conservation crisis,major Russian institutions collaborated in a dynamic program to develop and implement RTs for the sustainable management of amphibian biodiversity.An initial primary focus was the captive breeding of threatened Russian endemic anuran and caudate species,using RTs that varied from environmental manipulation to the use of exogenous gonadotropic hormones to stimulate reproduction.These species were mostly from Palearctic or cool mountain regions,but also included a wide range of species from warm regions.Other early achievements included the successful cry opreservation of anuran spermatozoa and anuran diploid pluripotent cell nuclei,in order to store both the matrilineal and patrilineal genomes in biobanks,with their subsequent development to the blastula stage after implantation into enucleated oocytes.After the turn of the 21st Century,in support of the priorities of the Amphibian Conservation Action Plan(2007),we developed RTs for the refrigerated storage of testicular or urinary spermatozoa for days to weeks at 4℃,the cryopreservation of urinary spermatozoa using a novel cryoprotectant,the in vitro fertilisation of hormonally induced oocytes either fresh or after refrigerated ex situ or in situ storage,and the artificial insemination of salamanders with fresh spermatozoa.In this article,we describe previously unpublished techniques and techniques from obscure Russian sources.