Short Communication: Effect of cryopreservation on ultrastructure and mitochondrial function of albino Pangasius catfish spermatozoa




Abstract. Hasanah U, Binawanto, Alimuddin A, Boediono A, Kursini E. 2020. Short Communication: Effect of cryopreservation on ultrastructure and mitochondrial function of albino Pangasius catfish spermatozoa. Biodiversitas 21: 4524-4528. Cryopreservation techniques have been carried out on many endangered species and animals with unique characteristics. Successful cryopreservation techniques vary between species depending on various factors. The study used cryopreserved spermatozoa of the albino Pangasius catfish as samples. The cryopreserved spermatozoa were analyzed by its ultrastructure, functional mitochondria, and viability. The cryopreservation was performed using a combination of 10% methanol, skim milk, and fish ringer extender. A deep freezer is used for cryopreservation at -80 °C with a storage period of 14 days. Observations were made on fresh spermatozoa, post-equilibration spermatozoa, and frozen-thaw spermatozoa. This study found there were differences in ultrastructure and morphology in the three treatments. Fresh spermatozoa and post-equilibration spermatozoa appeared intact membrane, mitochondrial, and flagellar structures. In contrast, in frozen-thaw spermatozoa, there was damage to the cell membrane. The study showed different percentages of yields on functional mitochondria of fresh spermatozoa (98 ± 2%), spermatozoa post-equilibration (57 ± 7%), frozen-thaw spermatozoa (42 ± 3.21%). Cell viability showed that there were differences in viability of fresh spermatozoa and frozen-thaw (p <0.05), the results of fresh spermatozoa (92 ± 0.57%) spermatozoa post-equilibration (80 ± 3.51%), frozen-thaw spermatozoa (61 ± 2.30%).The study concluded that the spermatozoa cryopreservation affects the ultrastructure, mitochondrial function, and viability in albino Pangasius catfish spermatozoa.


Bautista FEA., Junior ASV, Corcini CD, Acosta IB, Caldas SS, Primel, Zanette EGJ. 2018. The herbicide atrazine affects sperm quality and the expression of antioxidant and spermatogenesis genes in zebrafish testes. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology 206: 17-22.
Carton-Garcia F, Riesco MF, Cabrita E, Herráez MP, Robles V. 2013. Quantification of lesions in nuclear and mitochondrial genes of Sparus aurata cryopreserved sperm. Aquaculture 402: 106-112.
De Baulny BO, Le Vern Y, Kerboeuf D, Maisse G. 1997. Flow cytometric evaluation of mitochondrial activity and membrane integrity in fresh and cryopreserved rainbow trout (Oncorhynchus mykiss) spermatozoa. Cryobiology 34 (2): 141-149.
Evenson DP, Darzynkiewicz Z, Melamed MR. 1982. Simultaneous measurement by flow cytometry of sperm cell viability and mitochondrial membrane potential related to cell motility. Journal of Histochemistry & Cytochemistry 30(3): 279-280.
Graham JK, Kunze E, Hammerstedt RH. 1990. Analysis of sperm cell viability, acrosomal integrity, and mitochondrial function using flow cytometry. Biology of reproduction 43(1): 55-64.
Guo W, Shao J, Li P, Wu J, Wei Q. 2016. Morphology and ultrastructure of Brachymystax lenox tsinlingensis spermatozoa by scanning and transmission electron microscopy. Tissue and cell 48(4): 321-327.
Harvey B, Kelley RN, Ashwood-Smith MJ. 1982. Cryopreservation of zebra fish spermatozoa using methanol. Canadian Journal of Zoology 60(8) : 1867-1870.
He S, Woods C. 2004. Effects of dimethyl sulfoxide and glycine on cryopreservation induced damage of plasma membranes and mitochondria to striped bass (Morone saxatilis) sperm. Cryobiology 48(3): 254-262.
Horvath Á., Miskolczi E, Urbányi B. 2003. Cryopreservation of common carp sperm. Aquatic Living Resources 16(5): 457-460.
Iswadi MI, Ann ZF. 2016. Morphometry of head spermatozoa of sexually matured Malayan Gaur Bulls (Bos gaurus hubbacki). J. Trop. Agric and Fd. Sc 44(2): 315-324.
Islam MS, Akhter T. 2011. Tale of fish sperm and factors affecting sperm motility: a review. Advances in Life Sciences 1(1): 11-19.
Kopeika J, Zhang T, Rawson DM, Elgar G. 2005. Effect of cryopreservation on mitochondrial DNA of zebrafish (Danio rerio) blastomere cells. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis 570(1): 49-61.
Lahnsteiner F, Weismann T, Patzner RA. 1997. Methanol as cryoprotectant and the suitability of 1.2 ml and 5 ml straws for cryopreservation of semen from salmonid fishes. Aquaculture Research 28(6) : 471-479.
Lahnsteiner F. 2003. The spermatozoa and eggs of the cardinal fish. Journal of Fish Biology 62(1): 115-128.
Lahnsteiner F, Mansour N, Kunz FA. 2011. The effect of antioxidants on the quality of cryopreserved semen in two salmonid fish, the brook trout (Salvelinus fontinalis) and the rainbow trout (Oncorhynchus mykiss). Theriogenology 76(5): 882-890.
Madhavi M, Kailasam M, Mohanlal DL. 2015. Ultrastructure of sperm of the Spotted scat (Scatophagus argus, Linnaeus, 1766) observed by scanning and transmission electron microscopy. Animal reproduction science 153: 69-75.
Maricchiolo G, Genovese L, Laurà R, Micale V, Muglia U. 2004. Fine structure of spermatozoa in the common pandora (Pagellus erythrinus Linnaeus, 1758)(Perciformes, Sparidae). Histology and histopathology.
Moraes CR, Meyers S. 2018. The sperm mitochondrion: Organelle of many functions. Animal reproduction science 194: 71-80.
Perchec G, Jeulin C, Cosson J, Andre F, Billard R. 1995. Relationship between sperm ATP content and motility of carp spermatozoa. Journal of cell science 108(2): 747-753.
Tiersch TR, Goudie CA, Carmichael GJ. 1994. Cryopreservation of channel catfish sperm: storage in cryoprotectants, fertilization trials, and growth of channel catfish produced with cryopreserved sperm. Transactions of the American Fisheries Society 123(4) : 580-586.
Viveiros AT, Amaral TB, Orfao LH, Isau ZA., Caneppele D, Leal MC. 2011. Sperm cryopreservation of tiete tetra Brycon insignis (Characiformes): effects of cryoprotectants, extenders, thawing temperatures and activating agents on motility features. Aquaculture Research 42(6): 858-865.

Most read articles by the same author(s)

1 2 > >>