Cryopreservation of Prochilodus lineatus semen: effect of cryoprotectants combination
Abstract
The study was conducted with the objective of comparing the toxicity and the effect of the combination of intra and extracellular cryoprotectants in curimba Prochilodus lineatus sperm cells subjected to cryopreservation. Semen from 19 males were analyzed and diluted in four solutions comprised of intra and extracellular cryoprotectants in the following combinations: methanol + lactose, methanol + egg yolk, DMSO + lactose; and DMSO + egg yolk. A portion of the diluted semen was frozen while the remaining fraction was kept in repose and evaluated after 10 min. For freezing, the diluted samples were packaged in 0.50 mL straws and placed into liquid nitrogen vapor for 24 h and, after this time, submerged into liquid nitrogen for 10 days. The combination of DMSO + lactose was less toxic to the diluted semen, resulting in higher motility rates and durations when compared to the other treatments. After thawing, the highest motility rate and duration were obtained using lactose as extracellular cryoprotectant, regardless of its combination. There was no significant difference between treatments when analyzing sperm morphology after thawing. Considering the effects of the tested treatments, the use of lactose as an extracellular cryoprotectant added with DMSO or methanol is the most suitable, since these combinations presented the highest motility rates and durations and low morphological change rate after thawing.
References
Aquaculture International, 12(1): 69-85.
FRANCISCATTO, R.T.; MURGAS, L.D.S.; MILIORINI, A.B.; SILVA, M.O.B.; LOGATO, P.V.R. 2002 Qualidade do sêmen de curimba (Prochilodus lineatus) e taxa de fertilidade após resfriamento a 4 ºC. Revista Brasileira de Reprodução Animal, 26(3): 213-215.
GAO, D.Y.; LIU, J.; LIU, C.; MCGANN, L.E.;WATSON, P.F.; KLEINHANS, F.W.; MAZUR, P.; CRITSER, E.S.; CRITSER, J.K. 1995 Prevention of osmotic injury to human spermatozoa during addition and removal of glycerol. Human Reproduction, 10(5): 1109-1122.
HARVEY, B. and KELLEY, R.N. 1984 Chilled storage of Sarotherodon mossambicus milt. Aquaculture, 36(1-2): 85-95.
JAMIESON, B.G.M. 1991 Fish evolution and systematics: evidence from spermatozoa. Cambridge: Cambridge University Press. 319p.
KAVAMOTO, E.T.; BARNABE, V.H.; CAMPOS,B.E.S.; ANDRADE-TALMELLI, E.F. 1999 Anormalidades morfológicas nos espermatozóides do Curimbatá, Prochilodus scrofa (Steidachner, 1881) (Osteichthyes, Characiformes, Prochilodontidae). Boletim do Instituto de Pesca, 25(único): 61-66.
LI, P.; HULAK, M.; KOUBEK, P.; SULC, M.;DZYUBA, B.; BORYSHPOLETS, S.; RODINA, M;, GELA, D.; MANASKOVA-POSTLEROVA, P.; PEKNICOVA, J.; LINHART, O. 2010 Ice-age endurance: the effects of cryopreservation on proteins of sperm of common carp, Cyprinus carpio L. Theriogenology, 74(3): 413-423.
MARIA, N.A.; VIVEIROS, A.T.M.; FREITAS, R.T.F.; OLIVEIRA, A.V. 2006 Extenders and cryoprotectants for cooling and freezing of piracanjuba (Brycon orbignyanus) semen, an endangered Brazilian teleost fish. Aquaculture, 260(1): 298í 306.
MARTÍNEZ-PÁRAMO, S.; PÉREZ-CEREZALES, S.; GÓMEZ-ROMANO, F.; BLANCO, G.; SÁNCHEZ,
J.A.; HERRÁEZ, M.P. 2009 Cryobanking as tool for conservation of biodiversity: effect of brown
trout sperm cryopreservation on the male genetic potential. Theriogenology, 71(4): 594í 604.
MARTÍNEZ-PÁRAMO, S.; DIOGO, P.; DINIS, M.T.; HERRÁEZ, M.P.; SARASQUETE, C.; CABRITA,
E. 2012 Incorporation of ascorbic acid andαtocopherol to the extender media to enhance
antioxidant system of cryopreserved sea bass sperm. Theriogenology, 77(6): 1129-1136.
MILIORINI, A.B. 2006 Ativadores e concentrações de metanol e dimetilsulfóxido na qualidade do sêmen criopreservado de curimba (Prochilodus lineatus). Lavras. 99p. (Dissertação de Mestrado.
Universidade Federal de Lavras - UFLA).Available at: <http://repositorio.ufla.br/handle
/1/1884> Access on: 18 Nov. 2015.
MILIORINI, A.B.; MURGAS, L.D.S.; ROSA, P.V.;OBERLENDER, G.; PEREIRA, G.J.M.; COSTA, D.V. 2011 A morphological classification proposal for curimba (Prochilodus lineatus) sperm damages after cryopreservation. Aquaculture Research, 42(2): 177-187.
MORAES, G.V.; STREIT JR, D.P.; RIBEIRO, R.P.; SAKAGUTI, E.S.; SOUZA, E.D.; POVH, J.A. 2004
Ação de diferentes indutores reprodutivos hormonais no aparecimento de anormalidades
morfológicas em espermatozoides de piavuçu (Leporinus macrocephalus), curimbatá Prochilodus lineatus) e carpa comum (Cyprinus carpio). Boletim do Instituto de Pesca, 30(2): 109-116.
MURGAS, L.D.S.; MILIORINI, A.B.; FREITAS,R.T.F.; PEREIRA, G.J.M. 2007 Criopreservação
do sêmen de curimba (Prochilodus lineatus) mediante adição de diferentes diluidores,
ativadores e crioprotetores. Revista Brasileira de Zootecnia, 36(3): 526-531.
MURGAS, L.D.S.; FELIZARDO, V.O.; ANDRADE, E.S.; FERREIRA, M.R.; PAULA, D.A.J.;
CARVALHO, A.F.S. 2014 Cryopreservation of sperm in Brazilian migratory freshwater fish. In:
YAMASHIRO, H. Recent Advances in Cryopreservation. Rijeka: InTech. p.59-71.
NAVARRO, R.D.; OLIVEIRA, A.A.; FILHO, O.P.R.;CARRARA, F.P.; PEREIRA, F.K.S.; SANTOS,
L.C. 2007 Reprodução induzida de curimbatá (Prochilodus affinis) com uso de extrato bruto
hipofisário de rã touro (Rana catesbeiana). Zootecnia Tropical, 25(2): 143-147.
PAULINO, M.S.; MURGAS, L.D.S.; FELIZARDO, V.O.; FREITAS, R.T.F. 2012 Anormalidades espermáticas de Piaractus mesopotamicus após descongelamento utilizando diferentes
metodologias. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 64(6): 1591-1596.
RIBEIRO, R.I.M.A. and GODINHO, H.P. 2003 Criopreservação do semen testicular do teleósteo
"piau-açu†Leporinus macrocephalus. Arquivos Brasileiros de Medicina Veterinária e Zootecnia,
55(1): 75-79.
SADIQUL ISLAM, M. and AKHTER, T. 2011 Tale of fish sperm and factors affecting sperm motility:
a review. Advances in Life Sciences, 1(1): 11-19.
SQUIRES, E.L.; KEITH, S.L.; GRAHAM, J.K. 2004 Evaluation of alternative cryoprotectants for
preserving stallion spermatozoa. Theriogenology,62(6): 1056-1065.
