Hostname: page-component-cd9895bd7-8ctnn Total loading time: 0 Render date: 2024-12-27T08:47:21.906Z Has data issue: false hasContentIssue false

Insights into soy lecithin and egg yolk-based extenders for chilling canine spermatozoa

Published online by Cambridge University Press:  05 December 2018

Andressa Dalmazzo
Affiliation:
University of São Paulo, College of Veterinary Medicine and Animal Science, Department of Animal Reproduction, Cidade Universitária, São Paulo, Brazil
Daniel de Souza Ramos Angrimani
Affiliation:
University of São Paulo, College of Veterinary Medicine and Animal Science, Department of Animal Reproduction, Cidade Universitária, São Paulo, Brazil
João Diego A. Losano*
Affiliation:
University of São Paulo, College of Veterinary Medicine and Animal Science, Department of Animal Reproduction, Cidade Universitária, São Paulo, Brazil
Carolina C. Rocha
Affiliation:
University of São Paulo, College of Veterinary Medicine and Animal Science, Department of Animal Reproduction, Cidade Universitária, São Paulo, Brazil
Carlos A. B. Sobrinho
Affiliation:
University of São Paulo, College of Veterinary Medicine and Animal Science, Department of Animal Reproduction, Cidade Universitária, São Paulo, Brazil
João Rafael Chinait Gurgel
Affiliation:
University of São Paulo, College of Veterinary Medicine and Animal Science, Department of Animal Reproduction, Cidade Universitária, São Paulo, Brazil
Pedro Ivo Monteiro Pacheco
Affiliation:
University of São Paulo, College of Veterinary Medicine and Animal Science, Department of Animal Reproduction, Cidade Universitária, São Paulo, Brazil
Claudia Kiyomi Minazaki
Affiliation:
University of São Paulo, College of Veterinary Medicine and Animal Science, Department of Animal Reproduction, Cidade Universitária, São Paulo, Brazil
Silvia E. Crusco
Affiliation:
University of São Paulo, College of Veterinary Medicine and Animal Science, Department of Animal Reproduction, Cidade Universitária, São Paulo, Brazil
Marcilio Nichi
Affiliation:
University of São Paulo, College of Veterinary Medicine and Animal Science, Department of Animal Reproduction, Cidade Universitária, São Paulo, Brazil
Valquíria H. Barnabe
Affiliation:
University of São Paulo, College of Veterinary Medicine and Animal Science, Department of Animal Reproduction, Cidade Universitária, São Paulo, Brazil
*
Address for correspondence: João Diego A. Losano. University of São Paulo, College of Veterinary Medicine and Animal Science, Department of Animal Reproduction, Av. Prof. Dr. Orlando Marques de Paiva, n. 87, Cidade Universitária, São Paulo, SP, CEP 05508–270, Brazil. E-mail: jdalosano@usp.br

Summary

The aim of this study was to compare different concentrations of soy lecithin (LEC0.01%, LEC0.05% and LEC0.1%) with egg yolk (Control) in cooling extenders during the storage of semen at 5ºC for 5 days. Twelve dogs (n = 12) were selected, and semen was cooled and assessed after 2, 24, 48, 72, 96 or 120 h. At each time point, sperm were analyzed for kinetic patterns (using computer-assisted sperm analysis), mitochondrial activity (3′3- diaminobenzidine assay), lipid peroxidation (TBARS assay), DNA fragmentation (SCSA®) and plasma and acrosome membrane integrity (eosin/nigrosin and fast green/rose Bengal stains, respectively). The Control group (1814.4 ± 197.2) presented the highest rates of lipid peroxidation at 120 h. Conversely, progressive motility (42.8 ± 4%), linearity (45.4 ± 1%), and VAP (88 ± 3%) were higher in the Control group. In addition, there was lower mitochondrial activity in the Control group at 72 h. Therefore, our data show that lecithin used at these concentrations was not able to maintain sperm viability at as high qualities as would egg yolk. Moreover, the decrease in high mitochondrial activity and the persistence of sperm motility may indicate a compensatory mechanism in canine spermatozoa (i.e., glycolytic pathway). Furthermore, these higher lipid peroxidation indexes could indicate the necessity for future therapy using extenders and antioxidants over a long cooling time for dog sperm.

Type
Research Article
Copyright
© Cambridge University Press 2018 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Aitken, RJ and De Iuliis, GN (2010) On the possible origins of DNA damage in human spermatozoa. Mol Hum Reprod 16, 313.Google Scholar
Angrimani, DS, Lucio, CF, Veiga, GA, Silva, LC, Regazzi, FM, Nichi, M and Vannucchi, CI (2014) Sperm maturation in dogs: sperm profile and enzymatic antioxidant status in ejaculated and epididymal spermatozoa. Andrologia 46, 814819.Google Scholar
Angrimani, DS, Barros, PM, Losano, JD, Cortada, CN, Bertolla, RP, Guimaraes, MA, Correa, SH, Barnabe, VH and Nichi, M (2017) Effect of different semen extenders for the storage of chilled sperm in Tigrina (Leopardus tigrinus). Theriogenology 89, 146154.Google Scholar
Axner, E and Lagerson, E (2016) Cryopreservation of dog semen in a Tris extender with 1% or 2% soya bean lecithin as a replacement of egg yolk. Reprod Domest Anim 51, 262268.Google Scholar
Beccaglia, M, Anastasi, P, Chigioni, S and Luvoni, GC (2009a) Tris-lecithin extender supplemented with antioxidant catalase for chilling of canine semen. Reprod Domest Anim 44, 345349.Google Scholar
Beccaglia, M, Anastasi, P and Luvoni, GC (2009b) Freezing of canine semen in an animal-free protein extender. Vet Res Commun 33(Suppl 1), 7780.Google Scholar
Brito, MM, Lucio, CF, Angrimani, DS, Losano, JD, Dalmazzo, A, Nichi, M and Vannucchi, CI (2017) Comparison of cryopreservation protocols (single and two-steps) and thawing (fast and slow) for canine sperm. Anim Biotechnol 28, 6773.Google Scholar
Buege, JA and Aust, SD (1978) Microsomal lipid peroxidation. Methods Enzymol 52, 302310.Google Scholar
Crespilho, AM, Nichi, M, Guasti, PN, Freitas-Dell’Aqua, CP, Sa, MF, Maziero, RR, Dell’Aqua, JA and Papa, FO (2014) Sperm fertility and viability following 48 h of refrigeration: evaluation of different extenders for the preservation of bull semen in liquid state. Anim Reprod Sci 146, 126133.Google Scholar
Cross, NL (1994) Phosphatidylcholine enhances the acrosomal responsiveness of human sperm. J Androl 15, 484488.Google Scholar
Evenson, D and Jost, L (2000) Sperm chromatin structure assay is useful for fertility assessment. Methods Cell Sci 22, 169189.Google Scholar
Fariello, RM, Pariz, JR, Spaine, DM, Cedenho, AP, Bertolla, RP and Fraietta, R (2012) Association between obesity and alteration of sperm DNA integrity and mitochondrial activity. BJU Int 110, 863867.Google Scholar
Forouzanfar, M, Sharafi, M, Hosseini, SM, Ostadhosseini, S, Hajian, M, Hosseini, L, Abedi, P, Nili, N, Rahmani, HR and Nasr-Esfahani, MH (2010) In vitro comparison of egg yolk-based and soybean lecithin-based extenders for cryopreservation of ram semen. Theriogenology 73, 480487.Google Scholar
Fukui, Y, Kohno, H, Togari, T, Hiwasa, M and Okabe, K (2008) Fertility after artificial insemination using a soybean-based semen extender in sheep. J Reprod Dev 54, 286289.Google Scholar
Galvez, MJ, Ortiz, I, Hidalgo, M, Morrell, JM and Dorado, J (2015) Should single layer centrifugation of dog semen be done before or after the semen is cooled? Vet Rec 176, 359.Google Scholar
Gomez, E, Irvine, DS and Aitken, RJ (1998) Evaluation of a spectrophotometric assay for the measurement of malondialdehyde and 4-hydroxyalkenals in human spermatozoa: relationships with semen quality and sperm function. Int J Androl 21, 8194.Google Scholar
Hesser, A, Darr, C, Gonzales, K, Power, H, Scanlan, T, Thompson, J, Love, C, Christensen, B and Meyers, S (2017) Semen evaluation and fertility assessment in a purebred dog breeding facility. Theriogenology 87, 115123.Google Scholar
Hidalgo, M, Bayon, JC, Galvez, MJ, Urbano, M, Ortiz, I and Dorado, J (2014) Cryopreservation of dog semen using a soybean lecithin-based extender: effect on sperm viability and acrosome integrity. Reprod Domest Anim 49, 72.Google Scholar
Hrudka, F (1987) Cytochemical and ultracytochemical demonstration of cytochrome c oxidase in spermatozoa and dynamics of its changes accompanying ageing or induced by stress. Int J Androl 11, 457458.Google Scholar
Iguer-ouada, M and Verstegen, JP (2001a) Evaluation of the “Hamilton Thorn computer-based automated system” for dog semen analysis. Theriogenology 55, 733749.Google Scholar
Iguer-ouada, M and Verstegen, JP (2001b) Long-term preservation of chilled canine semen: effect of commercial and laboratory prepared extenders. Theriogenology 55, 671684.Google Scholar
Irvine, DS, Twigg, JP, Gordon, EL, Fulton, N, Milne, PA and Aitken, RJ (2000) DNA integrity in human spermatozoa: relationships with semen quality. J Androl 21, 3344.Google Scholar
Kasimanickam, VR, Kasimanickam, RK, Memon, MA and Rogers, HA (2012) Effect of extenders on sperm mitochondrial membrane, plasma membrane and sperm kinetics during liquid storage of canine semen at 5°C. Anim Reprod Sci 136, 139145.Google Scholar
Katz, DF, Yanagimachi, R and Dresdner, RD (1978) Movement characteristics and power output of guinea-pig and hamster spermatozoa in relation to activation. J Reprod Fertil 52, 167172.Google Scholar
Kmenta, I, Strohmayer, C, Muller-Schlosser, F and Schafer-Somi, S (2011) Effects of a lecithin and catalase containing semen extender and a second dilution with different enhancing buffers on the quality of cold-stored canine spermatozoa. Theriogenology 75, 10951103.Google Scholar
Lagergren, CG (1953) On the eosin-nigrosin stain and some other methods for the appraisal of sperm vitality with special reference to practical application. Ann Ostet Ginecol 75, 9981005.Google Scholar
Leemans, B, Gadella, BM, Stout, TA, Nelis, H, Hoogewijs, M and Van Soom, A (2015) An alkaline follicular fluid fraction induces capacitation and limited release of oviduct epithelium-bound stallion sperm. Reproduction 150, 193208.Google Scholar
Lopes, G, Simoes, A, Ferreira, P, Martins-Bessa, A and Rocha, A (2009) Differences in preservation of canine chilled semen using different transport containers. Anim Reprod Sci 112, 158163.Google Scholar
Losano, J, Angrimani, D, Dalmazzo, A, Rui, BR, Brito, MM, Mendes, CM, Kawai, G, Vannucchi, CI, Assumpcao, M, Barnabe, VH and Nichi, M (2017a) Effect of mitochondrial uncoupling and glycolysis inhibition on ram sperm functionality. Reprod Domest Anim 52, 289297.Google Scholar
Losano, JDA, Padin, JF, Mendez-Lopez, I, Angrimani, DSR, Garcia, AG, Barnabe, VH and Nichi, M (2017b) The stimulated glycolytic pathway is able to maintain ATP levels and kinetic patterns of bovine epididymal sperm subjected to mitochondrial uncoupling. Oxid Med Cell Longev 2017, 1682393.Google Scholar
Lucio, CF, Silva, LC, Regazzi, FM, Angrimani, DS, Nichi, M, Assumpcao, ME and Vannucchi, CI (2016) Effect of reduced glutathione (GSH) in canine sperm cryopreservation: In vitro and in vivo evaluation. Cryobiology 72, 135140.Google Scholar
Mellanby, RJ, Ogden, R, Clements, DN, French, AT, Gow, AG, Powell, R, Corcoran, B, Schoeman, JP and Summers, KM (2013) Population structure and genetic heterogeneity in popular dog breeds in the UK. Vet J 196, 9297.Google Scholar
Minervini, F, Guastamacchia, R, Pizzi, F, Dell’Aquila, ME and Barile, VL (2013) Assessment of different functional parameters of frozen–thawed buffalo spermatozoa by using cytofluorimetric determinations. Reprod Domest Anim 48, 317324.Google Scholar
Mukai, C and Okuno, M (2004) Glycolysis plays a major role for adenosine triphosphate supplementation in mouse sperm flagellar movement. Biol Reprod 71, 540547.Google Scholar
Nascimento, JM, Shi, LZ, Tam, J, Chandsawangbhuwana, C, Durrant, B, Botvinick, EL and Berns, MW (2008) Comparison of glycolysis and oxidative phosphorylation as energy sources for mammalian sperm motility, using the combination of fluorescence imaging, laser tweezers, and real-time automated tracking and trapping. J Cell Physiol 217, 745751.Google Scholar
Nichi, M, Goovaerts, IG, Cortada, CN, Barnabe, VH, De Clercq, JB and Bols, PE (2007) Roles of lipid peroxidation and cytoplasmic droplets on in vitro fertilization capacity of sperm collected from bovine epididymides stored at 4 and 34°C. Theriogenology 67, 334340.Google Scholar
Ohkawa, H, Ohishi, N and Yagi, K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95, 351358.Google Scholar
Pope, CE, Zhang, YZ and Dresser, BL (1991) A simple staining method for evaluating acrosomal status of cat spermatozoa. J Zoo Wildlife Med 22, 8795.Google Scholar
Reed, ML, Ezeh, PC, Hamic, A, Thompson, DJ and Caperton, CL (2009) Soy lecithin replaces egg yolk for cryopreservation of human sperm without adversely affecting postthaw motility, morphology, sperm DNA integrity, or sperm binding to hyaluronate. Fertil Steril 92, 17871790.Google Scholar
Rota, A, Strom, B and Linde-Forsberg, C (1995) Effects of seminal plasma and three extenders on canine semen stored at 4°C. Theriogenology 44, 885900.Google Scholar
Setyawan, EM, Kim, MJ, Oh, HJ, Kim, GA, Jo, YK, Lee, SH, Choi, YB and Lee, BC (2016) Spermine reduces reactive oxygen species levels and decreases cryocapacitation in canine sperm cryopreservation. Biochem Biophys Res Commun 479, 927932.Google Scholar
Sicherle, CC, Maia, MS, Bicudo, SD, Rodello, L and Azevedo, HC (2011) Lipid peroxidation and generation of hydrogen peroxide in frozen–thawed ram semen supplemented with catalase or Trolox. Small Rumin Res 95, 144149.Google Scholar
Toker, MB, Alcay, S, Gokce, E and Ustuner, B (2016) Cryopreservation of ram semen with antioxidant supplemented soybean lecithin-based extenders and impacts on incubation resilience. Cryobiology 72, 205209.Google Scholar