Sri Wahjuningsih, Khaeruddin Khaeruddin, Herlina Pratiwi, Aprilianna Putri Zahara Nafsina Luvita Sari, Yuli Arif Tribudi, Ani Atul Arif, Tatan Kostaman
The Gaga chicken, an indigenous Indonesian breed with distinctive vocal characteristics, is facing a population decline. Sperm cryopreservation is a potential method to preserve its genetic resources. This study aimed to evaluate the effects of different base diluents on calcium ion (Ca2+) concentration, malondialdehyde (MDA) levels, and sperm ultrastructure after freezing. Four diluents were tested: Ringer’s Lactate-egg yolk (RLEY), Ringer’s Acetate-egg yolk (RAEY), Ringer’s Lactate-skim milk (RLSM), and Ringer’s Acetate-skim milk (RASM). Semen was collected from male Gaga chickens using the massage method, diluted with each treatment, and packed into mini straws. The samples underwent equilibration at 5 °C for 2 h, pre-freezing for 10 min, freezing for 24 h, and thawing at 37 °C for 30 s. Results showed that RLEY produced the highest intracellular Ca2+ concentration post-thaw. MDA levels increased after freezing-thawing but were lower in RLEY than in RASM. Ultrastructural analysis revealed sperm head damage in all treatments except RLEY. Based on these findings, RLEY was more effective than RASM for preserving Gaga chicken sperm during cryopreservation, offering better protection against freezing-induced damage.Highlights sperm preservation of Gaga chicken is important to develop a breeding program the use of ringer diluent Ringer’s Lactate-egg yolk (RLEY) found to be effective as cryoprotectant. © 2025 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.
Department of Animal Science, Universitas Brawijaya, East Java, Malang, Indonesia; Program Study of Animal Science, Universitas Muhammadiyah Sinjai, South Sulawesi, Sinjai, Indonesia; Department of Animal Veterinary Science, Universitas Brawijaya, East Java, Malang, Indonesia; Department of Animal Production, Universitas Padjadjaran, West Java, Sumedang, Indonesia; Department of Animal Science, Universitas Tanjungpura, West Kalimantan, Pontianak, Indonesia; Center of Animal Husbandry, National Research and Innovation Agency (BRIN), Bogor, Indonesia
