The effects of cryoprotectants on chilled pirapitinga (Piaractus brachypomus) embryos at various ontogenetic stages
Resumo
Cryopreservation has not been successfully used to preserve fish embryos, although chilling techniques have been used with good results. The aim of this study was to chill Piaractus brachypomus embryos at different stages of development in some cryoprotectants and for various periods of chilling. Embryos at the following ontogenetic stages were used: blastoderm – 1.2 hours post-fertilization (hpf); epiboly – 5 hpf; blastopore closure – 8 hpf; and appearance of optic vesicle – 13 hpf. One hundred embryos were selected from each of the four stages and chilled in methanol, methylglycol or dimethylsulfoxide (DMSO) for 6, 8, 10 or 12 hours, at 2ºC. The total number of treatments was 4 stages x 3 cryoprotectants x 4 periods of chilling. The highest percentage of normal and live larvae (30.6%) was observed when embryos were chilled at 13-hpf in methanol for 6 hours. In general, larvae chilled at a more developed stages (8 and 13 hpf), in methanol and for shorter periods could survive chilling and develop normally, compared to the other treatments. Therefore, P. brachypomus embryos at the optical vesicle appearance stage (13 hpf) should be chilled in a solution containing 17.5% glucose and 10% methanol for up to eight six at 2°C.
Palavras-chave
Referências
AHAMMAD M.M.; BHATTACHARYYA D.; JANA B.B . The hatching of common carp (Cyprinus carpio L.) embryos in response to exposure to different concentrations of cryoprotectant at low temperatures. Cryobiology v.44, p. 114-121, 2002. doi: 10.1016/S0011-2240(02)00012-3
AHAMMAD M.M.; BHATTACHARYYA D.; JANA B.B. Stage-dependent hatching responses of rohu (Labeo rohita) embryos to different concentrations of cryoprotectants and temperatures. Cryobiology, v.46, p. 1-16, 2003. doi: 10.1016/S0011-2240(02)00138-4
ASAHINA E.; TAKAHASHI T. Cryopreservation of sea urchin embryos and sperm. Development, Growth and Differentiation, v. 21, p. 423-430. 1979. doi: 10.1111/j.1440-169X.1979.00423.x
BART A. New approaches in cryopreservation of fish embryos, In: CRYOPRESERVATION IN AQUATIC SPECIES. World Aquaculture Society, Baton Rouge, 2000. p.179-187.
BEGOVAC P.C.; WALLACE R.C. Major vitelline envelope proteins in piperfish oocytes originate within the follicle and are associated with the Z3 layer. Journal of Experimental Zoology. v.251, p. 56–73. 1989. doi: 10.1002/jez.1402510108
BEIRÃO J.; ROBLES V.; HERRAEZ M.P.; SARASQUETE C.; DINIS M.T.; CABRITA E. Cryoprotectant microinjection toxicity and chilling sensitivity in gilthead seabream (Sparus aurata) embryos. Aquaculture, v.261, p. 897-903, 2006. doi: 10.1016/j.aquaculture.2006.07.039
CABRITA E.; ROBLES V.; CHEREGUINI O.; WALLACE J.C.; HERRÁEZ M.P. Effect of different cryoprotectants and vitrificant solutions on the hatching rate of turbot embryos (Scophthalmus maximus). Cryobiology, v.47, p. 204-213, 2003. doi: 10.1016/j.cryobiol.2003.10.001
CABRITA E.; ROBLES V.; WALLACE J.C.; SARASQUETE M.C.; HERRÁEZ M.P. Preliminary studies on the cryopreservation of gilthead seabream (Sparus aurata) embryos. Aquaculture, v.251, p.245–255. 2006. doi: 10.1016/j.aquaculture.2005.04.077.
CHAO N.H.; LIN T.T.; CHEN Y.J.; HSU H.W.; ILIAO.C. Cryopreservation of late embryos and early larvae in the oyster and hard clam. Aquaculture, v. 155, p.31-44, 1997. doi: 10.1016/S0044-8486(97)00107-5
CHEN S.L.; TIAN Y.S. Cryopreservation of flounder (Paralichthys olivaceus) embryos by vitrification. Theriogenology, v. 63, p. 1207-1219, 2005. doi: 10.1016/j.theriogenology.2004.06.007
COSTA R.S.; VELARDI J.M.C.; SENHORINI J.A.; VERÍSSIMO-SILVEIRA R.; SILVEIRA A.N. Resfriamento de embriões de peixes neotropicais In: AQUACIÊNCIA, Palmas-TO, 2012.
DING F.H.; XIAO Z.Z.; LI J. Preliminary studies on the vitrification of red sea bream (Pagrus major) embryos. Theriogenology, v. 68, p. 702-708, 2007. doi: 10.1016/j.theriogenology.2007.05.064
DINNYÉS A.B.; URBÁNYI B.; BARANYAI I. Chilling sensitivity of carp (Cyprinus carpio) embryos at different developmental stages in the presence or absence of cryoprotectants: work in progress. Theriogenology, v. 50, p.1-13, 1998. doi: 10.1016/S0093-691X(98)00108-3
DOBRINSKY J.R. Advancements in cryopreservation of domestic animal embryos, Theriogenology, v. 57, p.285-302, 2002. doi: 10.1016/S0093-691X(01)00672-0
HAGA Y. On the subzero temperature preservation of fertilized eggs of rainbow trout. Nippon Suisan Gakkaishi v.48, p. 1569–1572. 1982. doi:10.2331/suisan.48.1569
FORNARI D.C.; RIBEIRO R.P.; STREIT D.P.; VARGAS L.; BARRERO N.M.L.; MORAES G.V. Freezing injuries in the embryos of Piaractus mesopotamicus. Zygote, v.19, p.345- 350. 2011. doi: 10.1017/S0967199410000432
HAGEDORN M,; KLEINHANS F.W.; FREITAS R.; LIU J.; HSU E.W.; WILDT D.E.; RALL W.F. Water distribution and permeability of zebrafish embryos, Brachydanio rerio. . Journal of Experimental Zoology. v. 278, p. 356–371.1997. doi: 10.1002/(SICI)1097-010X
HAGEDORN M.; PETERSON A.; MAZUR P.; KLEINHANS F.W. High ice nucleation temperature of zebrafish embryos: slow-cooling is not an option. Cryobiology v.49, p. 181–189.2004. doi: 10.1016/j.cryobiol.2004.07.001
HARVEY B. Cooling of embryonic cells, isolated blastoderms, and intact embryos of the zebra fish Brachydanio rerio to minus -196°C. Cryobiology v.20, p. 440–447.1983. doi: 10.1016/0011-2240(83)90034-2
HARVEY B.; ASHWOOD-SMITH M.J. Cryoprotectant penetration and supercooling in the eggs of Salmonid fish. Cryobiology v.19, p. 29–40. 1982. doi: 10.1016/0011-2240(82)90122-5
HOLT W.V. Basic aspects of frozen storage of semen. Animal Reproduction Science v.62, p. 3–22. 2000.doi: 10.1016/S0378-4320(00)00152-4
JAOUL A. ; ROUBAUD P. Resistance de l’oeuf de carp commune (Cyprinus carpio L. Cyprinidae) a des chocs thermiques chauds ou froids. Canadien Journal of Zoology, v. 60, p. 3409–3419.1982.doi: 10.1139/z82-431
LAHNSTEINER F. The effect of internal and external cryoprotectants on zebrafish (Danio rerio) embryos. Theriogenology v.69, p.384–396. 2008. doi: 10.1016/j.theriogenology.2007.10.007
LEIBO S.P. Sources of variation in cryopreservation. In: Cryopreservation in aquatic species. (ed by Tiersch T.R., Mazik P.M..), pp. 75–83. 2000. World Aquaculture Society, Baton Rouge.
LIU K.; CHOU T.; LIN H. Cryosurvival of goldfish embryos after subzero freezing. Aquatic Living Resources v.6, p. 145–153. 1993.doi: 10.1051/alr:1993007
LIU X.H.; ZHANG T.; RAWSON D.M. Feasibility of vitrification of zebrafish embryos using methanol. Cryoletters v.19, p. 309–318. 1998.doi: 10.1006/cryo.1996.0001
LOPES T.S.; ROMAGOSA E.; STREIT Jr D.P.; RIBEIRO R.P.; DIGMAYER M Cooling of pacu (Piaractus mesopotamicus) embryos at various stages of development for 6 or 10 hours. Theriogenology v.75, p.570–576. 2011.. doi: 10.1016/j.theriogenology.2010.09.030
LOPES T.S; STREIT Jr. D.P.; FORNARI D.C.; de OLIVEIRA D.; RIBEIRO R.P.; ROMAGOSA E. Chilling curves for Piaractus mesopotamicus (Holmberg, 1887) embryos stored at -8C. Zygote v.21, p. 345 – 350. 2012.doi: 10.1017/S0967199412000020
MADDOCK B.G. A technique to prolong the incubation period of brown trout ova. Progressive Fish-Culturist v.36, p 219–222. 1974.doi: 10.1577/1548-8659(1974)36[219:ATTPTI]2.0.CO;2
NASCIMENTO A.F.; MARIA A.N,; PESSOA N.O.; CARVALHO M.A.M.; VIVEIROS A.T. Out-of-season sperm cryopreserved in different media of the Amazonian freshwater fish pirapitinga (Piaractus brachypomus). Animal Reproduction Science v.118, p.324–329. 2010. doi: 10.1016/j.anireprosci.2009.07.002
OLIVE P.J.W.; WANG W.B. Cryopreservation of Nereis virens (polychaeta Annelida) larvae: the mechanism of cryopreservation of a differentiated metazoan. Cryobiology v.34, p. 284–294.1997. doi: 10.1006/cryo.1997.2006
ROBERTSON S.M.; LAWRENCE A.L.; NEILL W.; ARNOLD C.R.; McCARTHY G Toxicity of the cryoprotectants glycerol, dimethyl sulfoxide, ethylene glycol, methanol, sucrose, and sea salt solutions to the embryos of red drum. Progressive Fish-Culturist v.50, p. 148–154. 1988. doi: 10.1577/1548-8640(1988)050<0148:TOTCGD>2.3.CO;
ROBLES V.; CABRITA E.; REAL M.; ÁLVAREZ R.; HERRÁEZ M.P. Vitrification of turbot embryos: preliminary assays. Cryobiology v.47, p 30–39.2003. doi: 10.1016/S0011-2240(03)00066-X
ROUBAUD P. ; CHAILLOU C. ; SJAFEI D. Variations cycliques de la tolerance a un thermique froid appliqué au cours de la segmentation de l'embryon de la carpe commune (Cyprinus carpio L.). Canadien Journal of Zoology v.63,p. 657–663. 1985. doi: 10.1139/z85-094
VAL A.L.; VAL-ALMEIDA V.M.F. Fishes of the Amazon and their environment: physiological and biochemical aspects. Springer Heidelberg. 1995. cap.1, p.39-63.
STREIT Jr. D.P.; DIGMAYER M.; RIBEIRO R.P.; SIROL R.N.; MORAES G.V.; GALO J.M. Embriões de pacu submetidos a diferentes protocolos de resfriamento [Pacu embryos submitted to different cooling protocols]. Pesquisa Agropecuária Brasileira v.42, p. 1199–1202. 2007. doi: 10.1590/S0100-204X2007000800018
XIAO Z.Z.; ZHANG L.L.; XU X.Z.; LIU Q.H.; LI J.; MA D.Y.; XU S.H.; XUE Y.P.; XUE Q.Z. Effect of cryoprotectants on hatching rate of red seabream (Pagrus major) embryos. Theriogenology v.70, p. 1086–1092. 2008. doi: 10.1016/j.theriogenology.2008.06.028
ZHANG T.; LIU X.Z.; RAWSON D.M. Effects of methanol and developmental arrest on chilling injury in zebrafish (Danio rerio) embryo. Theriogenology v.59 p. 1545–1556. 2003. doi: 10.1016/S0093-691X(02)01199-8
ZHANG T.; RAWSON D.M .Studies on chilling sensitivity of zebrafish (Brachydanio rerio) embryos. Cryobiology v.32, p. 239–246. 1995. doi :10.1006/cryo.1995.1023
ZHANG T.; RAWSON D.M.; MORRIS G.J. Cryopreservation of pre-hatch embryos of zebrafish (Brachydanio rerio) embryos. Aquatic Living Resources v.6, p. 145–153. 1993.doi: 10.1051/alr:1993014
ZHANG Y.Z.; ZHANG S.C.; LIU X.Z.; XU Y.J.; HU J.; XU Y.Y.; LI J.; CHEN S.L. Toxicity and protective efficiency of cryoprotectants to flounder (Paralichthys olivaceus) embryos. Theriogenology v.63, p.763–773. 2005. doi: 10.1016/j.theriogenology.2004.04.011
Apontamentos
- Não há apontamentos.
Visitas