Animal Reproduction (AR)
https://animal-reproduction.org/article/doi/10.21451/1984-3143-AR2019-0067
Animal Reproduction (AR)
Original Article

Effect of supplementation of Aloe vera extracts in cold storage media and cryopreservation of domestic cat epididymal spermatozoa

Brenna de Sousa Barbosa; Fernanda Araujo dos Santos; Luãn Barbalho do Macêdo; Roberta Gonçalves Izzo; Denilsa Pires Fernandes; Érika Almeida Praxedes; Alexandre Rodrigues Silva; Marcelo Barbosa Bezerra

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Abstract

Abstract: This study evaluated the effect of the extract of Aloe vera at concentrations of 10% and 20% on the cryopreservation of sperm from the epididymis of domestic cats. Epididymal spermatozoa were recovered using the flotation technique and used in the treatments: control (TRIS-egg yolk at 20%), T10% (TRIS plus 10% of A. vera extract), and T20% (TRIS plus 20% of A. vera extract). The spermatozoa were subjected to 4ºC for 60 minutes, followed by 20 minutes in nitrogen vapors, and stored in a cryogenic cylinder. The samples were thawed at 37°C for 30 seconds. The sperm motility decreased (P<0.05) after thawing in the three treatments. Only the spermatozoa in the control treatment maintained post-thawing vigor. The viability of spermatozoa decreased in the treatments with A. vera (P<0.05). According to the hypoosmotic test, all treatments maintained the sperm membrane functionality (P>0.05) during freezing; however, after thawing, it decreased (P<0.05) in the T10% and T20% treatments. The morphology and chromatin condensation of spermatozoa did not differ, regardless of the treatments and time of evaluation (P>0.05). The effect of the crude A. vera extract was not satisfactory on the cryopreservation of epididymal spermatozoa of domestic cats after thawing; although the motility of spermatozoa was similar to that found with the use of egg yolk, and it presented maintenance of the chromatin integrity. However, it is necessary to understand the action of the substances present in A. vera with the feline spermatozoa, well as the standardization and adjustment of physicochemical characteristics aiming at the future application of the vegetal extract.

Keywords

freezing, tomcat, sperm, Aloe vera, ryoprotectant

References

Aguiar GV, Santos BMB, Cavalcante JMM, Rodrigues FRN, Salgueiro CCM, Nunes JF. Addition of Aloe vera to powder coconut water solution (ACP-101®) like cryoprotector of the goat semen cooled at 4°C. Ciênc Anim. 2012;22:292-5.

Arruda RP, Celeghini ECC, Alonso MA, Carvalho HF, Oliveira LZ, Nascimento J, Silva DF, Affonso FJ, Lemes KM, Jaime JD. Métodos de avaliação da morfologia e função espermática: momento atual e desafios futuros. Rev Bras Reprod Anim. 2011;35:145-51.

Axnér E, Forsberg C. Sperm Morphology in the Domestic Cat, and its Relation with Fertility: A Retrospective Study. Reprod Domest Anim. 2007;291(3):282-91. http://dx.doi.org/10.1111/j.1439-0531.2007.00780.x. PMid:17506807.

Bittencourt RF, Ribeiro AL Fo, Santos ADF, Chalhoub M, Alves SGG, Vasconcelos MF, Leandro EES, Guimarães JD. Utilização do teste hiposmótico para avaliar a eficácia de diferentes protocolos de criopreservação do sêmen caprino. Cienc Anim Bras. 2006;6:213-8.

Bousseau S, Brillard JP, Marguant-Le Guienne B, Guérin B, Camus A, Lechat M. Comparison of bacteriological qualities of various egg yolk sources and the in vitro and in vivo fertilizing potential of bovine semen frozen in egg yolk or lecithin based diluents. Theriogenology. 1998;50(5):699-706. http://dx.doi.org/10.1016/S0093-691X(98)00175-7. PMid:10734444.

Brusentsev E, Kizilova E, Mokrousova V, Kozhevnikova V, Rozhkova I, Amstislavsky S. Characteristics and fertility of domestic cat epididymal spermatozoa cryopreserved with two different freezing media. Theriogenology. 2018;110:148-52. http://dx.doi.org/10.1016/j.theriogenology.2017.12.038. PMid:29396043.

Buranaamnuay K. Protocols for sperm cryopreservation in the domestic cat: a review. Anim Reprod Sci. 2017;183:56-65. http://dx.doi.org/10.1016/j.anireprosci.2017.06.002. PMid:28629655.

CBRA. Manual para exame andrológico e avaliação de sêmen animal. 3ª ed. Belo Horizonte: Colégio Brasileiro de Reprodução Animal; 2013. 53 p.

Chatdarong K, Thuwanut P, Manee-in S, Lohachit C, Axner E. Effects of Thawing Temperature and Post‐thaw Dilution on the Quality of Cat Spermatozoa. Reprod Domest Anim. 2010;45(2):221-7. http://dx.doi.org/10.1111/j.1439-0531.2008.01218.x. PMid:19090816.

Chatdarong K. Retained fertilizing capability in cryopreserved feline spermatozoa. Reprod Domest Anim. 2017;52(Suppl 2):261-4. http://dx.doi.org/10.1111/rda.12855. PMid:27807913.

Cheuquemán C, Faúndez R, Sánchez R, Risopatrón J. Changes in sperm function and structure after freezing in domestic cat spermatozoa. Andrologia. 2018;50(9):e13080. http://dx.doi.org/10.1111/and.13080. PMid:30039626.

Cocchia N, Ciani F, El-Rass R, Russo M, Borzacchiello G, Esposito V, Montagnaro S, Avallone L, Tortora G, Lorizio R. Cryopreservation of feline epididymal spermatozoa from dead and alive animals and its use in assisted reproduction. Zygote. 2010;18(1):1-8. http://dx.doi.org/10.1017/S0967199409990256. PMid:19939333.

Di Domenico D, Pedroso EMSR, Teixeira PPM. Diferenças da célula espermática suína e crioprotetores: revisão. Nucl Anim. 2015;7:31-42. http://dx.doi.org/10.3738/1982.2278.1169.

Emerenciano KDM, Lima GL, Peixoto GCX, Silva MA, Oliveira MGC, De Paula VV, Silva AR. Recuperação de espermatozoides epididimários de gatos domésticos (Felis catus) utilizando soluções à base de tris ou água de coco em pó. Acta Vet. Bras. 2013;7:148-53.

Estakhr J, Javdan N. Spermatogenic activity of Aloe vera in adult male rats. Pharmacologyonline. 2011;2:886-9.

Farias CFA. Estudo da eficácia da Aloe vera como crioprotetor vegetal na refrigeração de espermatozoides epididimários de bovinos [trabalho de conclusão]. João Pessoa: Universidade Federal da Paraíba; 2017. Portuguese.

Figueroa E, Valdebenito I, Farias JG. Technologies used in the study of sperm function in cryopreserved fish spermatozoa. Aquacult Res. 2016;47(6):1691-705. http://dx.doi.org/10.1111/are.12630.

Filliers M, Rijsselaere T, Bossaert P, Zambelli D, Anastasi P, Hoogewijs M, Van Soom A. In vitro evaluation of fresh sperm quality in tomcats: a comparison of two collection techniques. Theriogenology. 2010;74(1):31-9. http://dx.doi.org/10.1016/j.theriogenology.2009.12.016. PMid:20219239.

Freitas VS, Rodrigues RAF, Gaspi FOG. Propriedades farmacológicas da Aloe vera (L.). Burm. F. Rev. Bras. Plant. Med. 2014;16(2):299-307. http://dx.doi.org/10.1590/S1516-05722014000200020.

García-Vázquez FA, Gadea J, Matás C, Holt WV. Importance of sperm morphology during sperm transport and fertilization in mammals. Asian J Androl. 2016;18(6):844-50. http://dx.doi.org/10.4103/1008-682X.186880. PMid:27624988.

Hermansson U, Axnér E. Epididymal and ejaculated cat spermatozoa are resistant to cold shock but egg yolk promotes sperm longevity during cold storage at 4ºC. Theriogenology. 2007;67(7):1239-48. http://dx.doi.org/10.1016/j.theriogenology.2007.01.008. PMid:17349687.

Howard J, Donoghue AM, Johnston LA, Wildt DE. Zona pellucida filtration of structurally abnormal spermatozoa and reduced fertilization in teratospermic cats. Biol Reprod. 1993;49(1):131-9. http://dx.doi.org/10.1095/biolreprod49.1.131. PMid:8353179.

Kalthur G, Adiga SK, Upadhya D, Rao S, Kumar P. Effect of cryopreservation on sperm DNA integrity in patients with teratospermia. Fertil Steril. 2008;89(6):1723-7. http://dx.doi.org/10.1016/j.fertnstert.2007.06.087. PMid:17953963.

Kamimura CF, Jacomini JO, Beletti ME. Alterações de cromatina em espermatozóides de ovinos e caprinos avaliadas por azul de toluidina e alaranjado de acridina. Cienc Agrotec. 2010;34(1):212-9. http://dx.doi.org/10.1590/S1413-70542010000100027.

Kunkitti P, Axnér E, Bergqvist AS, Sjunnesson Y. In vitro fertilization using frozen-thawed feline epididymal spermatozoa from corpus and cauda regions. Theriogenology. 2016;86(6):1403-8. http://dx.doi.org/10.1016/j.theriogenology.2016.04.085. PMid:27242180.

Kunkitti P, Chatdarong K, Suwimonteerabutr J, Nedumpun T, Johannisson A, Bergqvist AS, Sjunnesson Y, Axnér E. Osmotic tolerance of feline epididymal spermatozoa. Anim Reprod Sci. 2017;185:148-53. http://dx.doi.org/10.1016/j.anireprosci.2017.08.014. PMid:28847638.

Lima DBC, Silva TFP, Cortez AA, Pinto JN, Magalhães FF, Caldini BN, Silva LDM. Recovery of sperm after epididymal refrigeration from domestic cats using ACP-117c and Tris extenders. Arq Bras Med Vet Zootec. 2016;68(4):873-81. http://dx.doi.org/10.1590/1678-4162-8653.

Macente BI, Mansano CFM, Pereira MM, Martins MIM, Gioso MM, Savi PAP, Gutierrez RR. Congelação de espermatozoides epididimários de gatos utilizando o diluidor Botu-crio® após refrigeração por 24 h em contêiner de transporte de sêmen Botutainer®. Acta Vet. Bras. 2012;6:112-7.

Martins MIM, Justino RC. Criopreservação espermática em felinos: estado da arte. Rev. Bras. Reprod. Anim. 2015;39:129-35.

Melo CCS, Oliveira ÉCS, Ramos RP, Lima CF, Rodrigues AES, Guerra MMP. Renovação do diluidor Tris com gema de ovo ou Aloe vera sp. Na viabilidade do sêmen canino refrigerado a 5°C–resultados preliminares. Acta Vet. Bras. 2014;8:142-3.

Müller G, Martino-Andradea AJ, Santos AS, Reghelina AL, Garcia DM, Sant’Ana GR, Spercoski KM, Meyer KB, Torres SM, Silva VA Jr, Moraisa RN. Testicular testosterone: estradiol ratio in domestic cats and its relationship to spermatogenesis and epididymal sperm morphology. Theriogenology. 2012;78(6):1224-34. http://dx.doi.org/10.1016/j.theriogenology.2012.05.018. PMid:22898016.

Nava-Trujillo H, Hernández-Fernández A, Quintero-Moreno A. Integridad de la cromatina y forma de la cabeza del espermatozoide de toro: evaluación simultánea con la tinción de azul de toluidina. Rev. Cien. 2012;2:211-6.

Nejatzadeh-Barandozi F. Antibacterial activities and antioxidant capacity of Aloe vera. Bioorg Med Chem Lett. 2013;3(1):1-8. PMid:23870710.

Penfold LM, Jost L, Evenson DP, Wildt DE. Normospermic versus teratospermic domestic cat sperm chromatin integrity evaluated by flow cytometry and intracytoplasmic sperm injection. Biol Reprod. 2003;69(5):1730-5. http://dx.doi.org/10.1095/biolreprod.103.016089. PMid:12890738.

Pukazhenthi B, Noiles E, Pelican K, Donoghue A, Wildt D, Howard J. Osmotic Effects on Feline Spermatozoa from Normospermic versus Teratospermic Donors. Cryobiology. 2000;40(2):139-50. http://dx.doi.org/10.1006/cryo.2000.2233. PMid:10788313.

Pukazhenthi B, Pelican K, Wildt D, Howard J. Sensitivity of domestic cat (Felis catus) sperm from normospermic versus teratospermic donors to cold-induced acrosomal damage. Biol Reprod. 1999;61(1):135-41. http://dx.doi.org/10.1095/biolreprod61.1.135. PMid:10377041.

Rozkot M, Frydrychová S, Beneŝová N, Opletal L. Small molecules of natural origin as a source of antimicrobial agents in pig breeding–review. Res Pig Breed. 2013;7:26-33.

Rui BR, Angrimani DSR, Bicudo LC, Losano JDA, Nichi M, Pereira RJG. A fast, low‐cost and efficient method for the diagnosis of sperm DNA fragmentation in several species. Reprod Domest Anim. 2017;53(1):171-5. http://dx.doi.org/10.1111/rda.13087. PMid:29086436.

Santos JFP, Gomes ET, Siqueira AKM, Cardoso RCS. Andrologia e criopreservação de sêmen em cães. Rev Bras Reprod Anim. 2016;40:167-79.

Sieme H, Oldenhof H, Wolkers WF. Mode of action of cryoprotectants for sperm preservation. Anim Reprod Sci. 2016;69:2-5. http://dx.doi.org/10.1016/j.anireprosci.2016.02.004. PMid:26936658.

Silva LDM, Mota-Filho AC. Recuperação e conservação de espermatozoides epididimários de mamíferos. Acta Vet. Bras. 2012;6:1-8.

Silva SV, Guerra MMP. Efeitos da criopreservação sobre as células espermáticas e alternativas para redução das crioinjúrias. Rev. Bras. Reprod. Anim. 2011;35:370-84.

Souza ALP, Lima GL, Peixoto GCX, Silva AM, Oliveira MF, Silva AR. Use of Aloe Vera-based extender for chilling and freezing collared peccary (Pecari tajacu) semen. Theriogenology. 2016;85(8):1432-8. http://dx.doi.org/10.1016/j.theriogenology.2016.01.007. PMid:26830302.

Souza ALP, Lima GL, Silva AR. Alternativas para o aperfeiçoamento dos protocolos de criopreservação de sêmen de animais selvagens. Rev Bras Reprod Anim. 2014;38:98-102.

Sringnam S, Kitiyanant Y, Lewin LM, Saikhun K. Semen quality and chromatin condensation in domestic cat sperm during passage through the epididymis. Witthayasan Kasetsat Witthayasat. 2011;45:46-58.

Vargas WDE. Efecto de la adición de un surfactante natural (Aloe vera) al diluyente Triladyl® para crioconservación de semen bovino en toros reproductores de agso-genes, Quito-Pichincha (tesis). Latacunga: Universidad Técnica de Cotopaxi; 2012.

Villaverde AISB, Melo CM, Corrente JE, Papa FO, Lopes MD. Comparação entre dois métodos de coloração para análise morfológica e acrossomal de espermatozóides de gato doméstico (Felis catus). Cienc Anim Bras. 2008;9:686-92.
 

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