Reproduction Abstracts

Introduction: Currently, one-third of amphibians species are threatened with extinction and assisted reproductive technologies (ART) play an important role in their genetic management. The first step in developing ART is to create protocols for obtaining gametes. The aim of this study was to develop a protocol for hormonal induction of spermiation in the Natterjack toad (Bufo calamita).

Materials and methods: Males were divided into three groups (n=5/group) and each group was given an i.p. injection of 5, 10, or 15 IU hCG/g bodyweight. Spermic urine was collected from all males before the treatment and every hour post-treatment (up to 9 h) and sperm quality was assessed.

Results and discussion: None of the 15 males produced sperm before the hormone treatment. Results showed that all males treated with 10 and 15 IU/g of hCG released sperm, while only 40% of the males responded to 5 IU/g. There was no effect of hormone dose on sperm parameters, while time post-treatment did affect spermatozoa. The total percentage of moving sperm, percentage of sperm moving forward and quality of motility was higher from 1 to 4 h post-treatment with 10 IU hCG and from 1 to 6 h post-injection with 15 IU hCG (P≤0.044). Concentration of spermatozoa was lower in the first collection. Therefore, 15 IU of hCG should be used when sperm is going to be collected over a longer period of time for artificial fertilization. Here we describe for the first time a non-invasive protocol for spermatozoa collection from Bufo calamita, which may be important for future conservation efforts in this species.