Rebeca Marques Mascarenhas, Tarcízio Antonio Rego de Paula


The effect of the low intensity centrifugation (300xg), as well as the breed variation were evaluated. To this, semen of 12 dogs of the Beagle (n = 3), Schnauzer (n = 3), Doberman (n = 3) and Boxer (n = 3) breeds, 3 ejaculates collected per individual, totaling 36 ejaculates were used. The direct microscopic evaluation of the motility parameters and the hypoosmotic and supravital staining tests were used. The individual means of sperm vigor and motility varied from 3.5 to 5.0 and from 63 to 90% respectively, while the breed means ranged from 4.4 to 4.9 and from 80 to 90%, respectively. No significant difference was observed in any of the parameters evaluated in fresh or centrifuged semen of dogs among the different breeds studied. Centrifugation of semen resulted in a reduction of about 13% in sperm vigor and 14% in motility and sperm index. The analysis of the parameters of viability and sperm membrane integrity shows a 38% and 14% reduction in the percentage of reactive cells to the hypoosmotic test and supravital staining respectively. These results allow us to conclude that the centrifugation protocol, even at low intensity, significantly reduces sperm motility, the membrane integrity and sperm viability, this effect being more clearly perceived through the hypoosmotic test.



El efecto de la centrifugación a baja intensidad (300xg), así como la variación racial fueron evaluados. Para esto, semen de 12 perros de las razas Beagle (n=3), Schnauzer (n =3), Doberman (n = 3) y Boxer (n = 3), 3 eyaculados colectados por individuo, totalizando 36 eyaculados fueron usados. La medición por microscopia directa de los parámetros de motilidad, así como el test hipoosmótico y la tinción supravital fueron usadas. La media individual del vigor espermático y la motilidad variaron desde 3,5 a 5,0 y desde 63 a 90% respectivamente, mientras que las medias de las razas variaron desde 4,4 a 4,9 y desde 80 a 90%, respectivamente. No se observaron diferencias significativas en los parámetros evaluados el semen fresco y centrifugado entre las razas estudiadas. El análisis de los parámetros de vitalidad y la integridad de la membrana espermática muestra una reducción de 38% y 14% en el porcentaje de células reactivas al test hipoosmótico y a la tinción supravital respectivamente. Estos resultados llevan a la conclusión de que el protocolo de centrifugación, aun a baja intensidad, significativamente reduce la motilidad espermática y la integridad de la membrana y vitalidad espermática, siendo este efecto percibido más claramente a través del test hipoosmótico.

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Aquino-Cortez A., Pinheiro BQ., Lima DBC., Silva HVR., Mota-Filho AC., Martins JAM., Rodriguez-Villamil P., Moura AA., Silva LDM. (2017). Proteomic characterization of canine seminal plasma. Theriogenology 95: 178-186.

Aurich C. 2005. Factors affecting the plasma membrane function of cooled-stored stallion spermatozoa. Animal Reproduction. Science 89:65-75.

Batista M., Alamo D., Gonzáles F., Cruz MG. 2006. Garcia, A. Influence of freezing technique (Nitrogen liquid vs Ultrafreezer of -1520 C) and male-to-male variation over semen quality in Canarian Mastiff breed dogs. Reproduction in Domestic Animals 41: 423-428.

Bueno R., Costa E P., Guimarães JD., Valentim FM. 2001. Qualidade espermática de sêmen canino criopreservado. II - Utilização de dois protocolos de resfriamento. Arquivo Brasileiro de Medicina Veterinária e Zootecnia 53(3):372-379.

CBRA 1998. Manual para Exame Andrológico e Avaliação de Sêmen Animal.2.ed. Belo Horizonte: COLÉGIO BRASILEIRO DE REPRODUÇÃO ANIMAL, 1998. 49p.

Christensen BW., Asa CS., Wang C., Bauman K., Agnew MK., Lorton SP., Callahan M. 2013. Kinematic activity of gray wolf (Canis lupus) sperm in different extenders, added before or after centrifugation. Theriogenology 79: 953–960.

Cunha ICN., Lopes MD. 2009. Efeito da centrifugação sobre a qualidade do sêmen canino. Arquivo Brasileiro de Medicina Veterinária e Zootecnia 61(1):104-109.

Davis MJ., Pinto CR., Kozinsk DM., Minter LJ. 2007. Detection of reactive oxygen species in canine semen. Theriogenology 68: 492-518.

Ellerbrock RE., Prell MJ., Stewart JL., Bojko MS., Lima FS., Canisso IF. 2017. Comparison of Centrifugation and Noncentrifugation Methods to Cryopreserve Stallion Epididymal Semen. Journal of Equine Veterinary Science 50: 27-32.

Fontbonne A., Badinand F. 1993. Studies on freezing dog spermatozoa: effect of glycerol on motility after thawing. Journal of Reproduction and Fertility Suppl. 47:531-532.

Hermansson U., Ponglowhapan S., Linde-Forsberg C, Ström-Holst B. 2006. A short sperm–oocyte incubation time ZBA in the dog. Theriogenology 66:717-725.

Krause W., Viethen G. 1999. Quality assessment of computer-assisted semen analysis (CASA) in the andrology laboratory. Andrologia 31:125-129.

Koderle M., Aurich C., Schäfer-Somi S. 2009. The influence of cryopreservation and seminal plasma on the chromatin structure of dog spermatozoa. Theriogenology 72: 1215–1220.

Linde-Forsberg C., Forsberg M. 1989. Fertility in dogs in relation to semen quality and the time and site of insemination with fresh and frozen semen. Journal of Reproduction and Fertility Suppl. 39: 299-310.

Linde-Forsberg C., Strom Holst B., Govette G. 1999. Comparison of fertility data from vaginal vs intrauterine insemination of frozen-thawed dog semen: a retrospective study. Theriogenology 52:11-23.

Martins-Bessa A., Rocha A., Mayenco-Aguirre A. 2006. Comparing ethylene glycol with glycerol for cryopreservation of canine semen in egg-yolk TRIS extenders. Theriogenology 66:2047-2055.

Mascarenhas RM., Paula TAR., Matta SLP., Lanna LL., Fonseca CC., Neves MTD. 2006. Morfometria macro e microscópica e índices somáticos dos componentes testiculares de cães sem raça definida, da puberdade à senilidade. Revista CERES 53:106-112.

Matás C., Decuadro G., Martínez-Miró S., Gadea J. 2007. Evaluation of a cushioned method for centrifugation and processing for freezing boar semen. Theriogenology 67:1087-1091.

Morais RN., Mucciolo RG., Gomes MLF., Lacerda O., Moraes W., Moreira N., Graham LH., Swanson WF., Brown JL. 2002. Seasonal analysis of semen characteristics, serum testosterone and fecal androgens in the oncelot (Leopardus pardalis), margay (L. wiedii) and tigrina (L. tigrinus). Theriogenology 57:2027-2041.

Nöthling JO., Gerstenberg C., Volkmann DH. 1997.Semen quality after thawing: correlation with fertility and fresh semen quality in dogs. Journal of Reproduction and Fertility Suppl. 51:109-116.

Paula TAR., Cardoso FM. 1995. Alterações etárias na espermatogênese do cão. II. População celular dos túbulos seminíferos e rendimento espermatogênico. Arquivo Brasileiro de Medicina Veterinária e Zootecnia 47(4):535-547.

Peña AI., Barrio F., Quintela LA., Herradón PG. 1998. Effects of different glycerol treatments on frozen-thawed dog sperm longevity and acrosomal integrity. Theriogenology 50:63-174.

Peña AI., Lugilde LL., Barrio F., Herradón PG., Quintela LA. 2003. Effects of equex from different sources on post-thaw survival, longevity and intracellular Ca2+ concentration of dog spermatozoa. Theriogenology 59:1725-1739.

Peña A., Linde-Forsberg C. 2000. Effects of Equex, one- or two-step dilution, and two freezing and thawing rates on post-thaw survival of dog spermatozoa. Theriogenology 54:859-875.

Peña FJ., Núñez-Martínez I., Morán JM. 2006. Semen technologies in dog breeding: an update. Reproduction in Domestic Animals 41(Supp.2):21-29.

Prapaiwan N., Tharasanit T., Punjachaipornpol S., Yamtang D., Roongsitthichai A., Moonarmart W., Kaeoket K., Manee-in S. 2016. Low-density Lipoprotein Improves Motility and Plasma Membrane Integrity of Cryopreserved Canine Epididymal Spermatozoa. Asian-Australasian Journal of Animal Science 29(5):646-651.

Rappa KL., Rodriguez HF., Hakkarainen GC., Anchan RM., Mutter GL., Asghar W. 2016. Sperm processing for advanced reproductive technologies: Where are we today?. Biotechnology Advances 34:578–587.

Rijseelaere T., Van Soom A., Tanghe S., Coryn M., Maes D., Kruif A. 2005. New techniques for the assessment of canine semen quality: A review. Theriogenology 64:706-719.

Rijseelaere T., Van Soom A., Maes D., Kruif A. 2002. Effect of centrifugation on in vitro survival of fresh diluted canine spermatozoa. Theriogenology 57:1669-1681.

Rota A., Iguer-Ouada M., Verstegen J. 1999. Fertility after vaginal or uterine deposition of dog semen frozen in a Tris extender with or without Equex STM paste. Theriogenology 51:1045-1058.

Rota A., Stöm B., Linde-Forsberg C. 1995. Effects of seminal plasma and three extenders on canine semen stored at 4°C. Theriogenology 44:885-900.

Rota A., Milani C., Romagnoli S. 2007. Effect of post-thaw dilution with autologous prostatic fluid on dog semen motility and sperm acrosome status. Theriogenology 67:520-525.

Rota A., Milani C., Romagnoli S., Zucchini P., Mollo A. 2010. Pregnancy and conception rate after two intravaginal inseminations with dog semen frozen either with 5% glycerol or 5% ethylene glycol. Animal Reproduction Science 118:94-97.

Santos IW., Lima VFMH., Nisfeld LC., Ribeiro APC. 2003. Congelação do sêmen canino comparando diferentes concentrações de glicerol e diferentes tempos de equilíbrio. Archives of Veterinary Science 8(2):57-62.

Schelling C., Gaillard C., Dolf G. 2005. Genetic variability of seven dog breeds based on microsatellite markers. Journal of Animal Breeding and Genetics 122(Suppl 1):71-77.

Schrack J., Dolf G., Reichler IM., Schelling C. 2017. Factors influencing litter size and puppy losses in the Entlebucher Mountain dog. Theriogenology 95:163 170.

Sieme H., Knop K., Rath D. 2006. Effects of cushioned centrifugation on sperm quality in stallion semen stored cooled at 50C for 24h, and stored cooled for 2 or 24h and then frozen. Animal Reproduction Science 94:99-103.

Silva LDM., Onclin K., Lejeune B. 1996. Comparisons of intravaginal and intrauterine insemination of bitches with fresh or frozen semen. Veterinary Record 138:154-157.

Verstegen JP., Onclin K., Iguer-Ouada M. 2005. Long-term motility and fertility conservation of chilled canine semen using egg yolk added tris-glucose extender: In vitro an in vivo studies. Theriogenology 64: 720-733.

Vyt P., Maes D., Rijsselaere T., Dejonckheere E., Castryck F., Van Soom A. 2004. Motility Assessment of Porcine Spermatozoa: A Comparison of Methods. Reproduction in Domestic Animals 39: 447–453.

Yamashiro H., Narita K., Sugimura S., Han Y., Sugawara A., Morohaku K., Nakazato F., Konno T., Yoshida M., Soto E. 2007. Trehalose enhanced the freezability of Poodle dog sperm collected by artificial vagina. Animal Reproduction Science 102: 165-171.

Zhang W., Yi K., Chen C., Hou X., Zhou X. 2012. Application of antioxidants and centrifugation for cryopreservation of boar spermatozoa. Animal Reproduction Science 132: 123– 128.

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