Animal Reproduction (AR)
https://animal-reproduction.org/article/doi/10.1590/1984-3143-AR2023-0004
Animal Reproduction (AR)
ORIGINAL ARTICLE

Canine sperm vitrification with nonpermeable cryoprotectants and coconut water extender

Anton Antonov; Boyana Ivanova

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Abstract

This study was aimed to assess the efficiency of coconut water extender with addition of soy lecithin and sucrose as nonpermeable cryoprotectants for canine semen vitrification, using a simple method that yields a high survival rate of spermatozoa for clinical use. Twelve ejaculates from 12 adult normozoospermic dogs were collected separately by digital manipulation and only the second semen fraction was used in this study. After evaluation of volume, concentration, viability, total and progressive motility, velocity parameters and morphology, semen was diluted with a coconut water extender (50% (v/v(volume per volume)) coconut water, 25% (v/v) distilled water and 25% (v/v) 5% anhydrous monosodium citrate solution) with addition of soy lecithin and fructose at 1% and 0.25M sucrose until final concentration of 100x106 spermatozoa/ml. After equilibration at 5ºC for 60 minutes, semen was vitrified by “direct dropping method” into liquid nitrogen in spheres with a volume of 30 μl. After a week of storage the spheres were devitrified as three of them were dropped into 0.5 mL of CaniPlus AI medium (Minitüb, Germany), which was previously warmed in a water bath at 42ºC for 2 minutes and evaluated about the above mentioned parameters. It was found that vitrification resulted in a lower percentage of viable sperms, normal morphology, total and progressive motilities (p<0.05), but most of velocity parameters (VCL, VSL, VAP, LIN, ALH and BCF) did not differ (p>0.05) compared to fresh semen samples. In conclusion, our results demonstrate that vitrification with coconut water extender with addition of 1% soy lecithin and 0.25M sucrose as cryoprotectants, has an excellent potential for routine canine sperm cryopreservation.

Keywords

dog sperm, vitrification, lecithin, sucrose, coconut water

References

Abdel-Aziz Swelum A, Saadeldin IM, Ba-Awadh H, Al-Mutary MG, Moumen AF, Alowaimer AN, Abdalla H. Efficiency of commercial egg yolk-free and egg yolk-supplemented tris-based extenders for dromedary camel semen cryopreservation. Animals (Basel). 2019;9(11):999. http://dx.doi.org/10.3390/ani9110999 PMid:31752406.

Amirat-Briand L, Bencharif D, Vera-Munoz O, Pineau S, Thorin C, Destrumelle S, Desherces S, Anton M, Jouan M, Shmitt E, Tainturier D. In vivo fertility of bull semen following cryopreservation with an LDL (low density lipoprotein) extender: preliminary results of artificial inseminations. Anim Reprod Sci. 2010;122(3-4):282-7. http://dx.doi.org/10.1016/j.anireprosci.2010.09.006. PMid:20961718.

Antonov A, Ivanova B. Effect of equilibration process on canine spermatozoa after vitrification using coconut water extender with addition of soy lecithin and sucrose as nonpermiable cryoprotectants. Spermova. 2022;12(1):27-32. http://dx.doi.org/10.18548/aspe/0010.05.

Baiee F, Al-Wahab BA, Almusawi AA, Yu LL, Fitri WN, Haron AW, Bustani GS. Effect of vitrification on spermatozoa quality in bull semen. Eurasia J Biosci. 2020;14:3897-904.

Bustani GS, Baiee FH. Semen extenders: an evaluative overview of preservative mechanisms of semen and semen extenders. Vet World. 2021;14(5):1220-33. http://dx.doi.org/10.14202/vetworld.2021.1220-1233. PMid:34220124.

Cardoso RCS, Silva AR, Silva LDM. Comparison of two dilution rates on canine semen quality after cryopreservation in a coconut water extender. Anim Reprod Sci. 2006;92(3-4):384-91. http://dx.doi.org/10.1016/j.anireprosci.2005.05.028. PMid:16005585.

Cardoso RCS, Silva AR, Silva LDM. Use of the powdered coconut water (ACP-106®) for cryopreservation of canine spermatozoa. Anim Reprod. 2005;2(4):257-62. PMid:17214766.

Cardoso RCS, Silva AR, Uchoa DC, Silva LDM. Cryopreservation of canine semen using a coconut water extender with egg yolk and three different glycerol concentrations. Theriogenology. 2003;59(3-4):743-51. http://dx.doi.org/10.1016/S0093-691X(02)01151-2. PMid:12517378.

Caturla-Sánchez E, Sánchez-Calabuig M, Pérez-Gutiérrez JF, Cerdeira J, Castaño C, Santiago-Moreno J. Vitrification of dog spermatozoa: effects of two cryoprotectants (sucrose or trehalose) and two warming procedures. Cryobiology. 2018;80:126-9. http://dx.doi.org/10.1016/j.cryobiol.2017.11.001. PMid:29126865.

Concannon PW, Battista M. Canine semen freezing and artificial insemination. In: Kirk RW, editor. Current veterinary therapy X: small animal practice. Philadelphia: WB Saunders; 1989. p. 1247–59.

Curry MR. Cryopreservation of semen from domestic livestock. Rev Reprod. 2000;5(1):46-52. http://dx.doi.org/10.1530/ror.0.0050046. PMid:10711735.

Domosławska A, Zduńczyk S, Niżański W, Janowski T. Assessment of semen quality in infertile dogs using computer-assisted sperm analysis by the Hamilton-Thorne Semen Analyser. Bull Vet Inst Pulawy. 2013;57(3):429-32. http://dx.doi.org/10.2478/bvip-2013-0074.

Eilts BE. Theoretical aspects of canine semen cryopreservation. Theriogenology. 2005;64(3):692-7. http://dx.doi.org/10.1016/j.theriogenology.2005.05.019. PMid:15955551.

Enciso M, López-Fernández C, Fernández JL, García P, Gosálbez A, Gosálvez J. A new method to analyze boar sperm DNA fragmentation under bright-field or fluorescence microscopy. Theriogenology. 2006;65(2):308-16. http://dx.doi.org/10.1016/j.theriogenology.2005.05.044. PMid:15996725.

England GCW. Semen evaluation, artificial insemenation. In: Etinger JS, Feldman EC, editors. Veterinary internal medicine. Philadelphia, USA: WB Saunders Company; 2000. p. 1571-85.

Fiser PS, Fairfull RW, Hansen C, Panich PL, Shrestha JNB, Underhill L. The effect of warming velocity on motility and acrosomal integrity of boar sperm as influenced by the rate of freezing and glycerol level. Mol Reprod Dev. 1993;34:190-5. http://dx.doi.org/10.1002/mrd.1080340211. PMid: 8442956.

Galarza DA, Landi G, Mejía E, Samaniego JX, Mendez S, Soria ME, Taboada J, Sanchez-Calabuig MJ, Castano C, Santiago-Moreno J. Cryopreservation of dog epididymal spermatozoa by conventional freezing or ultra-rapid freezing with nonpermeable cryoprotectant. Cryobiology. 2021;103:15-21. http://dx.doi.org/10.1016/j.cryobiol.2021.10.002. PMid:34715114.

Gharajelar SN, Sadrkhanloo RA, Onsori M, Saberivand A. A comparative study on the effects of different cryoprotectants on the quality of canine sperm during vitrification process. Vet Res Forum. 2016;7(3):235-9. PMid:27872720.

Gunawan IWNF, Kardena IM, Suatha IK, Puja IK. Coconut water based extender effects on motility, viability, and DNA integrity of chilled kintamani dog semen. Veterinary Science and Medicine Journal. 2016;4:17-21.

Holt WV. Fundamental aspects of sperm cryobiology: the importance of species and individual differences. Theriogenology. 2000;53(1):47-58. http://dx.doi.org/10.1016/S0093-691X(99)00239-3. PMid:10735061.

Isachenko E, Isachenko V, Sanchez R, Katkov I, Kreienberg R. Cryopreservation of spermatozoa: old routine and new perspectives. In: Donnez J, editor. Principles and practice of fertility preservation. Cambridge: University Press; 2011. p. 176–198. http://dx.doi.org/10.1017/CBO9780511921896.017.

Isachenko V, Isachenko E, Katkov I, Montag M, Dessole S, Nawroth F, Van Der Ven H. Cryoprotectant-free cryopreservation of human spermatozoa by vitrification and freezing in vapor: effect on motility, DNA integrity, and fertilization ability. Biol Reprod. 2004;71(4):1167-73. http://dx.doi.org/10.1095/biolreprod.104.028811. PMid:15175233.

Kim S, Lee Y, Yang H, Kim Y-J. Rapid freezing without cooling equilibration in canine sperm. Anim Reprod Sci. 2012;130(1-2):111-8. http://dx.doi.org/10.1016/j.anireprosci.2011.12.014. PMid:22277841.

Layek SS, Mohanty TK, Kumaresan A, Parks JE. Cryopreservation of bull semen: evolution from egg yolk based to soybean based extenders. Anim Reprod Sci. 2016;172:1-9. http://dx.doi.org/10.1016/j.anireprosci.2016.04.013. PMid:27509873.

Linde-Forsberg C, Forsberg M. Fertility in dogs in relation to semen quality and the time and site of insemination with fresh or frozen semen. J Reprod Fertil. 1989;39:299-310. PMid:2621732.

Magnotti C, Cerqueira V, Lee‐Estevez M, Farias JG, Valdebenito I, Figueroa E. Cryopreservation and vitrification of fish semen: a review with special emphasis on marine species. Rev Aquacult. 2018;10(1):15-25. http://dx.doi.org/10.1111/raq.12145.

Mantena SK, Jagadish, Badduri SR, Siripurapu KB, Unnikrishnan MK. In vitro evaluation of antioxidant properties of Cocos nucifera Linn. water. Nahrung. 2003;47(2):126-31. http://dx.doi.org/10.1002/food.200390023 PMid:12744292.

Mazur P, Seki S. Survival of mouse oocytes after being cooled in a vitrification solution to -196°C at 95° to 70,000°C/min and warmed at 610° to 118,000°C/min: a new paradigm for cryopreservation by vitrification. Cryobiology. 2011;62(1):1-7. http://dx.doi.org/10.1016/j.cryobiol.2010.10.159. PMid:21055397.

Merino O, Risopatrón J, Sánchez R, Isachenko E, Figueroa E, Valdebenito I, Isachenko V. Fish (Oncorhynchus mykiss) spermatozoa cryoprotectant-free vitrification: stability of mitochondrion as criterion of effectiveness. Anim Reprod Sci. 2011;124(1-2):125-31. http://dx.doi.org/10.1016/j.anireprosci.2011.02.023. PMid:21392903.

Nunes JF, Sales FGM. El agua de coco (Cocus nucifera) in natura integral y adicionada con citoquinina, comodiluidor de semen caprino. Rev Ciencia Anim. 1993;3(3):273-81.

Ogbu CC, Ugwu SO, Ezebili CV. Use of coconut milk as an extender for cock semen. Res Opin Anim Vet Sci. 2014;4(10):571-7.

Partyka A, Niżański W, Ochota M. Methods of assessment of cryopreserved semen. In: Katkov II, editor. Current frontiers in cryobiology. Rijeka: In Tech Open Access; 2012. http://dx.doi.org/10.5772/33565.

Peña AI, Barrio F, Quintela LA, Herradón PG. Effect of different glycerol treatments on frozen-thawed dog sperm longevity and acrosomal integrity. Theriogenology. 1998;50(1):163-74. http://dx.doi.org/10.1016/S0093-691X(98)00122-8. PMid:10734483.

Peterson K, Kappen MA, Ursem PJ, Nothling JO, Colenbrander B, Gadella BM. Microscopic and flow cytometric semen assessment of Dutch AI-bucks: effect of semen processing procedures and their correlation to fertility. Theriogenology. 2007;67(4):863-71. http://dx.doi.org/10.1016/j.theriogenology.2006.11.003. PMid:17126897.

Pipan MZ, Casal ML, Šterbenc N, Virant Klun I, Mrkun J. Vitrification using soy lecithin and sucrose: a new way to store the sperm for the preservation of canine reproductive function. Animals (Basel). 2020;10(4):653. http://dx.doi.org/10.3390/ani10040653. PMid:32283781.

Puja IK, Sawitri NM, Maharani N, Gunawan IWNF, Heryani LGSS. A comparative study on the effects of coconut water based extenders on the quality of kintamani dog semen preserved at 4OC. Adv Anim Vet Sci. 2018;6(5):192-6. http://dx.doi.org/10.17582/journal.aavs/2018/6.5.192.196.

Rosato MP, Iaffaldano N. Cryopreservation of rabbit semen: comparing the effects of different cryoprotectants, cryoprotectant-free vitrification, and the use of albumin plus osmoprotectants on sperm survival and fertility after standard vapor freezing and vitrification. Theriogenology. 2013;79(3):508-16. http://dx.doi.org/10.1016/j.theriogenology.2012.11.008. PMid:23218394.

Sánchez R, Risopatrón J, Schulz M, Villegas J, Isachenko V, Kreinberg R, Isachenko E. Canine sperm vitrification with sucrose: effect on sperm function. Andrologia. 2011;43(4):233-41. http://dx.doi.org/10.1111/j.1439-0272.2010.01054.x. PMid:21486402.

Sansone G, Nastri MJF, Fabbrocini A. Storage of buffalo (Bubalus bubalis) semen. Anim Reprod Sci. 2000;62(1-3):55-76. http://dx.doi.org/10.1016/S0378-4320(00)00154-8. PMid:10924820.

Seeram NP, Adams LS, Henning SM, Niu Y, Zhang Y, Nair M, Heber D. In vitro antiproliferative, apoptotic and antioxidant activities of punicalagin, Ellagic acid and a total pomegranate tannin extract are enhanced in combination with other polyphenols as found in pomegranate juice. J Nutr Biochem. 2005;16(6):360-7. http://dx.doi.org/10.1016/j.jnutbio.2005.01.006. PMid:15936648.

Shah D, Rasappan, Shila, Karthik G. A simple method of human sperm vitrification. MethodsX. 2019;6:2198-204. http://dx.doi.org/10.1016/j.mex.2019.09.022. PMid:31667120.

Silva P, Bamunuarachchi A. Manufacture of carbonated tender coconut water and development of a process for the utilization of coconut flesh. Asian J Food Agro-Ind. 2009;2:210-3.

Simons N. Collection and cryopreservation of epididymal spermatozoa in dogs [dissertation]. Belgium: Faculty of Veterinary Medicine, Ghent University; 2018.

Ström B, Rota A, Linde-Forsberg C. In vitro characteristics of canine spermatozoa subjected to two methods of cryopreservation. Theriogenology. 1997;48(2):247-56. http://dx.doi.org/10.1016/S0093-691X(97)84072-1. PMid:16728124.

Tezcan F, Gültekin-Özgüven M, Diken T, Ozcelik B, Erim FB. Antioxidant activity and total phenolic, organic acid and sugar content in commercial pomegranate juices. Food Chem. 2009;115(3):873-7. http://dx.doi.org/10.1016/j.foodchem.2008.12.103.

Verstegen J, Iguer–Ouada M, Onclin M. Computer assisted semen analyzers in andrology research and veterinary practice. Theriogenology. 2002;57(1):149-79. http://dx.doi.org/10.1016/S0093-691X(01)00664-1. PMid:11775967.

Yong JWH, Ge L, Ng YF, Tan SN. The chemical composition and biological properties of coconut (Cocos nucifera L.) water. Molecules. 2009;14(12):5144-64. http://dx.doi.org/10.3390/molecules14125144. PMid:20032881.
 


Submitted date:
01/06/2023

Accepted date:
04/27/2023

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