Molecular Fish Sexing on Taisho Sanshoku Koi (Cyprinus carpio) Based on ArS.9-15 Gene Amplification using PCR Method
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Taisho Sanshoku is a variant of Koi fish (Cyprinus carpio) that has high demand due to its high economic value and relatively expensive price. This study aimed to determine the sex of the Taisho Sanshoku Koi fish by molecular sexing using the PCR method to amplify the ArS.9-15 gene. This study was initiated by rearing a 4–6 month-old of 10 Taisho Sanshoku Koi fish in a fish tank with a filter and oxygen aeration. The fish were fed with fish pellets for 1–3 days. The Koi fishes were then anesthetized using Koi anesthesia containing β-hydroxyethyl phenyl ether. Each fish's peripheral blood was collected as much as 0.5 mL per fish and then stored in tubes containing Ca-EDTA anticoagulant. The genomic DNA was extracted from peripheral blood samples and used as a template DNA for PCR amplification targeting ArS.9-15 gene. Agarose with 1.5% concentration and CybrSafe staining was used in electrophoresis for visualization of the PCR results then visualized in a dark chamber using a UV transilluminator. The Taisho Sanshoku Koi fish's sex was determined using descriptive analysis based on the electrophoresis results. According to the PCR results, the female Taisho Sanshoku Koi fish only produced one 800 bp DNA band, whereas the male fish produced two 800 bp and 1,100 bp DNA bands. The outcome of molecular fish sexing of the 10 Taisho Sanshoku Koi fish reported that 60% were male and 40% were female.
Andrian, K. N., ’Aisy, N. R., Novindasari, B. B. M., Nurrahmi, I. A., Santi, M. D., & Haryanto, A. (2023). Random Amplified Polymorphic DNA-Polymerase Chain Reaction Analysis of Four Koi Fish (Cyprinus carpio var. koi) Variants from Yogyakarta, Indonesia. IOP Conference Series: Earth and Environmental Science, 1174, 1–7.
Ariyanti, Y., & Sianturi, S. (2019). Ekstraksi DNA total dari sumber jaringan hewan (Ikan Kerapu) menggunakan metode kit for animal tissue. 3(December 2018), 40–45.
Aysi, N. R., Santi, M. D., Andrian, K. N., & Haryanto, A. (2022). Molecular fish sexing on Kohaku Koi (Cyprinus carpio) based on ArS.9-15 gene amplification by PCR method. IOP Conference Series: Earth and Environmental Science, 976(1), 1–6.
Budd, A., Banh, Q., Domingos, J., & Jerry, D. (2015). Sex Control in Fish: Approaches, Challenges and Opportunities for Aquaculture. Journal of Marine Science and Engineering, 2015, 329–355.
Chen, J., Wang, Y., Yue, Y., Xia, X., Du, Q., & Chang, Z. (2009). A novel male-specific DNA sequence in the common carp, Cyprinus carpio. Molecular and Cellular Probes, 23(5), 235–239.
de Kock, S., & Gomelsky, B. (2015). Japanese Ornamental Koi Carp: Origin, Variation and Genetics. In Biology and Ecology of Carp. CRC Press. pp: 27–53.
Falaye, A. E., Abah, A., & Sule, S. O. (2024). Effect of Gum Arabic (Acacia senegal) on Growth Performance, Carcass Quality and Health of Clarias gariepinus Juveniles. Jurnal Medik Veteriner, 7(1), 163–176.
Fikri, F., Wardhana, D. K., Purnomo, A., Khairani, S., Chhetri, S., & Purnama, M. T. E. (2022). Aerolysin gene characterization and antimicrobial resistance profile of Aeromonas hydrophila isolated from milkfish (Chanos chanos) in Gresik, Indonesia. Veterinary World, 15(7), 1759–1764.
Jaya, I., & Iqbal, M. (2009). Development of a Technique for Early Sexing of Koi (Ornamental Carp). Jurnal Ilmu-Ilmu Perairan Dan Perikanan Indonesia, 16(1), 7–15.
Lee, P. Y., Costumbrado, J., Hsu, C. Y., & Kim, Y. H. (2012). Agarose gel electrophoresis for the separation of DNA fragments. Journal of Visualized Experiments, 62, 1–5.
Li, M., Xu, H., Xu, W., Zhou, Q., Xu, X., Zhu, Y., Zheng, W., Li, W., Pang, Z., & Chen, S. (2020). Isolation of a Male-Specific Molecular Marker and Development of a Genetic Sex Identification Technique in Spotted Knifejaw (Oplegnathus punctatus). Marine Biotechnology, 22(4), 467–474.
Lind, C. E., Safari, A., Agyakwah, S. K., Attipoe, F. Y. K., El-Naggar, G. O., Hamzah, A., Hulata, G., Ibrahim, N. A., Khaw, H. L., Nguyen, N. H., Maluwa, A. O., Zaid, M., Zak, T., & Ponzoni, R. W. (2015). Differences in sexual size dimorphism among farmed tilapia species and strains undergoing genetic improvement for body weight. Aquaculture Reports, 1, 20–27.
Liu, H., Pang, M., Yu, X., Zhou, Y., Tong, J., & Fu, B. (2018). Sex-specific markers developed by next-generation sequencing confirmed an XX/XY sex determination system in bighead carp (Hypophthalmichthys nobilis) and silver carp (Hypophthalmichthys molitrix). DNA Research, 25(3), 257–264.
Martínez, P., Viñas, A. M., Sánchez, L., Díaz, N., Ribas, L., & Piferrer, F. (2014). Genetic architecture of sex determination in fish: Applications to sex ratio control in aquaculture. Frontiers in Genetics, 5(SEP), 1–13.
Mei, J., & Gui, J. F. (2015). Genetic basis and biotechnological manipulation of sexual dimorphism and sex determination in fish. Science China Life Sciences, 58(2), 124–136.
Monticini, P. (2010). The Ornamental Fish Trade Production and Commerce of Ornamental Fish: technical-managerial and legislative aspects. In GLOBEFISH Research Programme, 102, 134.
Nie, Z., Lü, P., Zhang, R., Tu, Y., Liu, Z., Li, Y., Tang, C., Li, X., Zhao, K., Zhou, Q., Li, F., Wang, J., Zeng, Z., Tu, M., & Zhang, H. (2021). A simple and rapid method for fish sex identification based on recombinase-aided amplification and its use in Cynoglossus semilaevis. Scientific Reports, 11(1), 1–11.
Tahir, N. D. M., Matori, M. F., & Gan, H. M. (2024). First Report of Aeromonas schubertii Infection in Striped Snakehead Channa striata (Bloch, 1793) Fingerlings in Malaysia. Jurnal Medik Veteriner, 7(1), 33–40.
Wasko, A. P., Martins, C., Oliveira, C., & Foresti, F. (2003). Non-destructive genetic sampling in fish. An improved method for DNA extraction from fish fins and scales. Hereditas, 138, 161–165.
Xing, T. F., Li, Y. L., & Liu, J. X. (2021). Female-specific genomic regions and molecular sex identification of the clearhead icefish (Protosalanx hyalocranius). BMC Genomics, 22(1), 1–9.
Yu, Y., Hilsdorf, A. W. S., Zhou, L., Lin, Q., & Gao, Z. X. (2022). Editorial: Genetics and molecular breeding in aquaculture animals. In Frontiers in Genetics, 13.
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