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Animal reproduction2019; 16(2); 340-347; doi: 10.21451/1984-3143-AR2019-0039

Can we use LDL instead of egg yolk in BotuCrio® extender to cryopreserve sperm from the Mangalarga Marchador stallion?

Abstract: The objective of this study was to compare the BotuCrio extender with the Merk - egg yolk and the INRA 82 modified by the inclusion of acetamide, methyl cellulose and trehalose in substitution of glycerol for freezing equine semen. The semen was diluted after centrifugation to obtain 100 x 10 of sperm/ml in: BotuCrio (control); Merk - egg yolk or INRA 82 modified (Experiment 1). The extended semen was packaged in 0.5 ml straws, cooled and frozen in a freezing machine. The control extender was superior in preserving the motility, VCL, VSL, VAP, LIN, STR and the BCF when compared to the Merk - egg yolk and INRA 82 modified (P < 0.05). The BotuCrio® preserved more effectively the equine sperm viability characteristics evaluated in Experiment 1 and was used as a control extender in Experiment 2 to test the effectiveness of using LDL in replacement of egg yolk. BotuCrio was superior to preserve progressive motility, VCL, VSL, VAP, LIN, STR and the percentage of functional integrity of sperm membranes compared to BotuCrio LDL (P 0.05). The fertility rate after AI with frozen semen in BotuCrio LDL was 37.5%.
Publication Date: 2019-10-23 PubMed ID: 33224296PubMed Central: PMC7673584DOI: 10.21451/1984-3143-AR2019-0039Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research aimed to examine the efficiency of using LDL (low-density lipoprotein), instead of egg yolk, in the BotuCrio® extender to preserve sperm from Mangalarga Marchador stallions. The study confirmed that while both LDL and the traditional egg yolk technique maintained similar overall sperm health, the traditional BotuCrio® extender showed superiority concerning specific sperm mobility and percentages of functional sperm membrane integrity.

Context and Methodology

  • The research was primarily focused on identifying better methods to preserve stallion sperm, specifically from the Mangalarga Marchador breed.
  • The extender compounds used were BotuCrio®, Merk – egg yolk, and a modified INRA 82. Out of these, BotuCrio® was marked as the control.
  • The comparative study was divided into two experiments; Experiment 1 compared BotuCrio®, Merk – egg yolk, and the modified INRA 82, while experiment 2 compared the effectiveness of BotuCrio® with LDL in place of egg yolk.
  • The semen was diluted, extended, packaged, cooled, and frozen in a freezing machine for assessment.

Findings of Experiment 1

  • The traditional BotuCrio® extender was found to be more effective than both Merk – egg yolk and the modified INRA 82 in preserving several features of the semen.
  • These features included sperm motility, VCL (track speed), VSL (straight line velocity), VAP (path velocity), LIN (linearity), STR (straightness), and BCF (beat cross frequency).
  • The success of BotuCrio® in Experiment 1 led to its use as a control in Experiment 2.

Findings of Experiment 2

  • When LDL was used as an alternative to egg yolk in BotuCrio®, it showed similar results in maintaining total sperm motility, ALH (amplitude of lateral head displacement), BCF, and the structural integrity of the membranes.
  • However, the traditional BotuCrio® was still superior in preserving progressive sperm motility, VCL, VSL, VAP, LIN, STR, and the percentage of functional integrity of sperm membranes.
  • The fertility rate with semen frozen in BotuCrio® LDL was found to be 37.5%.

Conclusion

  • The study suggests while LDL can maintain overall sperm health similarly as the egg yolk technique, the traditional BotuCrio® extender is superior in maintaining specific mobility characteristics and a higher percentage of functional sperm membrane integrity.

Cite This Article

APA
Snoeck PPDN, Pessoa THO, Pereira MGS, Bastos ICL, de Melo MIV. (2019). Can we use LDL instead of egg yolk in BotuCrio® extender to cryopreserve sperm from the Mangalarga Marchador stallion? Anim Reprod, 16(2), 340-347. https://doi.org/10.21451/1984-3143-AR2019-0039

Publication

ISSN: 1984-3143
NlmUniqueID: 101272804
Country: Brazil
Language: English
Volume: 16
Issue: 2
Pages: 340-347

Researcher Affiliations

Snoeck, Paola Pereira das Neves
  • Universidade Estadual de Santa Cruz, Ciências Agrárias e Ambientais, Ilheus, Bahia, Brazil.
Pessoa, Tainá Hortência Oliveira
  • Universidade Estadual de Santa Cruz, Ciências Agrárias e Ambientais, Ilheus, Bahia, Brazil.
Pereira, Marcelo Gonçalves Sousa
  • Universidade Estadual de Santa Cruz, Ciências Agrárias e Ambientais, Ilheus, Bahia, Brazil.
Bastos, Inara Cristina Leal
  • Universidade Estadual de Santa Cruz, Ciências Agrárias e Ambientais, Ilheus, Bahia, Brazil.
de Melo, Maria Isabel Vaz
  • Pontífica Universidade Católica de Minas Gerais, Medicina Veterinária, Belo Horizonte, Brazil.

References

This article includes 47 references
  1. Alvarenga MA, Graham JK, Keith SL, Landim-Alvarenga FC, Squires EL. Alternative cryoprotectors for freezing stallion spermatozoa. Abstracts of the 14th International Congress on Animal Reproduction; 2000; Stockholm, Sweden. Stockholm: ICAR; 2000. p. 157.
  2. Alvarenga MA, Leão KM, Papa FO, Graham Jk. Improvement of stallion semen post-thaw motility with the utilization of dimethyl-formamide as cryoprotector. Theriogenology 2002;57:459.
  3. Alvarenga MA, Leão KM, Papa FO, Landim-Alvarenga FC, Medeiros ASL, Gomes GM. The use of alternative cryoprotectors for freezing stallion semen. Proceedings of the Workshop on Transporting Gametes and Embryos; 2003; Massachusetts, USA. USA: Havemeyer Foundation; 2003. pp. 74–76.
  4. Alvarenga MA, Papa FO, Landim-Alvarenga FC, Medeiros AS. Amides as cryoprotectants for freezing stallion semen: a review.. Anim Reprod Sci 2005 Oct;89(1-4):105-13.
  5. Álvarez C, Gil L, González N, Olaciregui M, Luño V. Equine sperm post-thaw evaluation after the addition of different cryoprotectants added to INRA 96® extender.. Cryobiology 2014 Aug;69(1):144-8.
  6. Amirat L, Tainturier D, Jeanneau L, Thorin C, Gérard O, Courtens JL, Anton M. Bull semen in vitro fertility after cryopreservation using egg yolk LDL: a comparison with Optidyl, a commercial egg yolk extender.. Theriogenology 2004 Apr 1;61(5):895-907.
    pubmed: 14757475doi: 10.1016/s0093-691x(03)00259-0google scholar: lookup
  7. Avanzi BR, Ramos Rdos S, Araujo GH, Fioratti EG, Trinca LA, Dell'Aqua JA Jr, Melo E Oña CM, Zahn FS, Martin I, Alvarenga MA, Papa FO. Fixed-time insemination with frozen semen in mares: is it suitable for poorly fertile stallions?. Theriogenology 2015 Jun;83(9):1389-93.
  8. Bergeron A, Crête MH, Brindle Y, Manjunath P. Low-density lipoprotein fraction from hen's egg yolk decreases the binding of the major proteins of bovine seminal plasma to sperm and prevents lipid efflux from the sperm membrane.. Biol Reprod 2004 Mar;70(3):708-17.
    pubmed: 14613896doi: 10.1095/biolreprod.103.022996google scholar: lookup
  9. Bencharif D, Amirat-Briand L, Garand A, Anton M, Schmitt E, Desherces S, Delhomme G, Langlois ML, Barrière P, Destrumelle S, Vera-Munoz O, Tainturier D. Freezing canine sperm: comparison of semen extenders containing Equex and LDL (Low Density Lipoproteins).. Anim Reprod Sci 2010 Jun;119(3-4):305-13.
  10. Candeias ML, Alvarenga MA, Carmo MT, Ferreira HN, Maior MRS, Torres Filho RA, Rodrigues ALR, Brandão FZ. Semen cryopreservation protocols of Mangalarga Marchador stallions. R Bras Zootec 2012;41:1989–1995.
  11. CBRA. Manual for Andrological Examination and Evaluation of Animal Semen. 3rd. Belo Horizonte: CBRA; 2013. 104. Colégio Brasileiro de Reprodução Animal. 2013.
  12. Costa DNM, Silva DAM, Boakari YL, Ferreira SB, Branco MAC, Souza JAT. Efficiency of tris extenders and Botu-crio® on seminal parameters in stallion breeds Quarter Horse and Mangalarga Marchador. Cienc Anim Bras 2014;15:322–329.
  13. Dalimata AM, Graham JK. Cryopreservation of rabbit spermatozoa using acetamide in combination with trehalose and methyl cellulose.. Theriogenology 1997 Oct 1;48(5):831-41.
    pubmed: 16728175doi: 10.1016/s0093-691x(97)00305-1google scholar: lookup
  14. Gibb Z, Morris LH, Maxwell WM, Grupen CG. Dimethyl formamide improves the postthaw characteristics of sex-sorted and nonsorted stallion sperm.. Theriogenology 2013 Apr 15;79(7):1027-33.
  15. Gomes GM, Papa F O, Jacob JCF, Macedo LP, Leão KM, Machado MS, Alvarenga MA. Improvement of frozen-thawed spermatic parameters with utilization of MP-50 extender to Mangalarga Machador stallions. Rev Bras Reprod Anim 2002;26:187–189.
  16. Graham JK. Evaluation of alternative cryoprotectants for preserving stallion spermatozoa. Abstracts of the 14th International Congress on Animal Reproduction; 2000; Stockholm, Sweden. Stockholm: ICAR; 2000. p. 307.
  17. Hanada A, Nagase H. Cryoprotective effects of some amides on rabbit spermatozoa.. J Reprod Fertil 1980 Sep;60(1):247-52.
    pubmed: 7431324doi: 10.1530/jrf.0.0600247google scholar: lookup
  18. Harrison RA, Vickers SE. Use of fluorescent probes to assess membrane integrity in mammalian spermatozoa.. J Reprod Fertil 1990 Jan;88(1):343-52.
    pubmed: 1690300doi: 10.1530/jrf.0.0880343google scholar: lookup
  19. Henry M, Snoeck PPN, Cottorello ACP. Post-thaw spermatozoa plasma membrane integrity and motility of stallion semen frozen with different cryoprotectants. Theriogenology 2002;58:245–248.
  20. Hoffmann N, Oldenhof H, Morandini C, Rohn K, Sieme H. Optimal concentrations of cryoprotective agents for semen from stallions that are classified 'good' or 'poor' for freezing.. Anim Reprod Sci 2011 May;125(1-4):112-8.
  21. Jiang ZL, Li QW, Li WY, Hu JH, Zhao HW, Zhang SS. Effect of low density lipoprotein on DNA integrity of freezing-thawing boar sperm by neutral comet assay.. Anim Reprod Sci 2007 Jun;99(3-4):401-7.
  22. Juliani G, Henry M, Melo MIV. Freezing of equine semen in extenders with low density lipoproteins. Abstracts of the 15th International Congress on Animal Reproduction; 2004; Bahia, Brazil. Brazil: ICAR; 2004. p. 498.
  23. Magistrini M, Couty I, Palmer E. Interactions between sperm packaging, gas environment, temperature and diluent on fresh stallion sperm survival.. Acta Vet Scand Suppl 1992;88:97-110.
    pubmed: 1285546
  24. Manjunath P, Nauc V, Bergeron A, Ménard M. Major proteins of bovine seminal plasma bind to the low-density lipoprotein fraction of hen's egg yolk.. Biol Reprod 2002 Oct;67(4):1250-8.
    pubmed: 12297543doi: 10.1095/biolreprod67.4.1250google scholar: lookup
  25. Martin JC, Klug E, Günzel AR. Centrifugation of stallion semen and its storage in large volume straws.. J Reprod Fertil Suppl 1979;(27):47-51.
    pubmed: 289825
  26. Medeiros ASL, Gomes GM, Carmo MT, Papa FO, Alvarenga MA. Cryopreservation of stallion using different amides. Theriogenology 2002;58:273–276.
  27. Melo CM, Zahn FS, Martin I, Orlandi C, Dell’Aqua Jr JA, Alvarenga MA, Papa FO. Influence of semen storage and cryoprotectant on post-thaw viability and fertility of stallion spermatozoa. J Equine Vet Sci 2007;27:171–175.
  28. Melo MIV, Henry M. Hypoosmotic test for the evaluation of equine semen. Arq Bras Med Vet Zootec 1999;51:71–78.
  29. Melo MIV, Resende YF, Rezende EP, Snoeck PPN, Gomes DML, Carvalho TTC, Silva AVD, Neves MM, Bastos DG, Henry MRJM. Low density lipoproteins and egg yolk to preserve cooled donkey semen. Rev Bras Med Vet 2012;34:288–294.
  30. Moreno D, Bencharif D, Amirat-Briand L, Neira A, Destrumelle S, Tainturier D. Preliminary results: the advantages of low-density lipoproteins for the cryopreservation of equine semen. J Equine Vet Sci 2013;33:1068–1075.
  31. Moussa M, Marinet V, Trimeche A, Tainturier D, Anton M. Low density lipoproteins extracted from hen egg yolk by an easy method: cryoprotective effect on frozen-thawed bull semen.. Theriogenology 2002 Apr 1;57(6):1695-706.
    pubmed: 12035979doi: 10.1016/s0093-691x(02)00682-9google scholar: lookup
  32. Moustacas VS, Zaffalon FG, Lagares MA, Loaiza-Eccheverri AM, Varago FC, Neves MM, Heneine LG, Arruda RP, Henry M. Natural, but not lyophilized, low density lypoproteins were an acceptable alternative to egg yolk for cryopreservation of ram semen.. Theriogenology 2011 Jan 15;75(2):300-7.
  33. Neves MM, Heneine LGD, Henry M. Cryoprotection effectiveness of low concentrations of natural and lyophilized LDL (low density lipoproteins) on canine spermatozoa. Arq Bras Med Vet Zootec 2014;66:769–777.
  34. Oliveira RA, Rubin MIB, Silva CAM. Pregnancy rates using frozen semen of Criollo stallions with glycerol or dimethylformamid as cryoprotectants. Cienc Anim Bras 2013;14:488–494.
  35. Papa FO, Zahn FS, Dell’Aqua Jr JA, Alvarenga MA. Using the MP 50 extender for cryopreservation of equine semen. Rev Bras Reprod Anim 2002;26:184–187.
  36. Pillet E, Batellier F, Duchamp G, Furstoss V, Vern YLE, Kerboeuf D, Vidament M, Renseigné N. Freezing stallion semen in INRA96® based extender improves fertility rates in comparison with INRA82. Dairy Sci Technol 2008;88:257–265.
  37. Pillet E, Duchamp G, Batellier F, Beaumal V, Anton M, Desherces S, Schmitt E, Magistrini M. Egg yolk plasma can replace egg yolk in stallion freezing extenders.. Theriogenology 2011 Jan 1;75(1):105-14.
  38. Ramires Neto C, Monteiro GA, Sancler-Silva YFR, Papa P, Guasti PN, Resende HL, Papa FO, Dell’Áqua Jr JA, Alvarenga MA. Comparison of different freezing extenders for semen cryopreservation from stallions with poor and good semen freezability. J Equine Vet Sci 2014;34:58–60.
  39. Rossi TC, Papa FO, Santos TB, Macedo LP, Alvarenga MA, Melo CM, Dell’Aqua Junior JA. Efeito da utilização de diferentes crioprotetores e suas associações no processo de congelação de sêmen eqüino com meio MP50. Rev Bras Reprod Anim 2003;27:350–352.
  40. Silva MC, Moura LCO, Melo MIV, Mambrini JVM, Neves MM, Henry MRJM, Snoeck PPN. Prolonged post cooling but not pre-cooling equilibrium lengh improves the viability of ram sperm cryopreserved in an extender containing low-density lipoproteins. Small Ruminant Res 2014;119:88–95.
  41. Snoeck PPN, Cottorello ACP, Henry M. Viability and fertility of stallion semen frozen with ethylene glycol and acetamide as a cryogenic agent. Anim Reprod 2012;9:33–39.
  42. Snoeck PPN, Henry M, Melo MIV. Effect of differents freezing extenders on post thaw equine spermatozoa viability. Arq Bras Med Vet Zootec 2007;59:56–64.
  43. Snoeck PPN, Moura LCO, Silva MC, Machado-Neves M, Melo MIV, Heneine LGD, Henry M. Effect of storage conditions on the LDL effectiveness in ovine sperm cryopreservation.. Cryobiology 2017 Apr;75:88-90.
  44. Squires EL, Keith SL, Graham JK. Evaluation of alternative cryoprotectants for preserving stallion spermatozoa.. Theriogenology 2004 Sep 15;62(6):1056-65.
  45. Terraciano PB, Bustamante – Filho IC, Miquelito LV, Arlas TR, Castro F, Mattos RC, Passos EP, Oberst ER, Lima EOC. Cryopreservation of equine spermatozoa comparing different freezing rates combined with commercial extenders: laboratorial analysis. Cienc Rural 2008;38:1972–1977.
  46. Varela Junior AS, Corcini CD, Ulguim RR, Alvarenga MV, Bianchi I, Corrêa MN, Lucia T Jr, Deschamps JC. Effect of low density lipoprotein on the quality of cryopreserved dog semen.. Anim Reprod Sci 2009 Oct;115(1-4):323-7.
  47. Vidament M, Daire C, Yvon J M., Doligez P, Bruneau B, Magistrini M, Ecot P. Motility and fertility of stallion semen frozen with glycerol and/or dimethyl formamide. Theriogenology 2002;58:249–251.

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