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Post-cooling survival, growth and deformity rates in zebrafish embryos (Danio rerio)

  • Maria do Carmo Faria Paes (a1) and Laura Satiko Okada Nakaghi (a2)

Summary

This study investigated and analysed survival, growth and macro- and microscopic damage during the development of zebrafish embryos up to the adult stage after undergoing cooling. The embryos at 50% epiboly stage were selected, submerged in cryoprotectant solution of methanol and sucrose, cooled gradually to 0 ± 2°C temperature, and divided into two groups with different storage times (6 and 18 h). Subsequently, the embryos were reheated, rehydrated and incubated normally. The experiment lasted 5 months and, from hatching onward, the larvae were examined, collected and processed at pre-established time intervals. The hatching rate was significantly higher for the larvae stored for 18 h compared with the 6-h group. However, embryos from this group gave rise to a larger number of malformations, and these were much more severe compared with those in the 6 h group, which led to a higher mortality in the long term. Regarding larval length, the animals of the 6 h group had higher mean total length compared with the 18 h group, but both treatments were inferior to the control. Numerous macro- and microscopic malformations were observed and, in both treatments, only the morphologically normal individuals were able to develop to the adult stage, with organ development similar to the control, except for the gonads that were still undifferentiated in treated animals.

Copyright

Corresponding author

All correspondence to: Maria do Carmo Faria Paes. Department of Animal Morphology and Physiology, São Paulo State University, Campus Jaboticabal, São Paulo, Brazil. Access way Professor Paulo Donato Castellane, S/N, Jaboticabal-SP, CEP 14884–900, Brazil. Tel:/Fax: +55 16 3209 2654 (ext. 232). E-mail: maria.paes@fcav.unesp.br

References

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Ahammad, M.M., Bhattacharyya, D. & Jana, B.B. (2003). Hatching of common carp (Cyprinus carpio L.) embryos stored at 4 and –2°C in different concentrations of methanol and sucrose. Theriogenology 60, 1409–22.
Campbell, T.W. (2004). Hematology of Fish. In Thrall, M.A. (ed.) Veterinary Hematology and Clinical Chemistry. Lippincott Williams & Wilkins, pp. 277–89.
Coelho, E.B. (2014). Mechanisms of Edema Formation. Medical symposium: Semiology, Ribeirão Preto, 37, 189–98.
Costa, J.M. (2015). Phosphorus levels in diets for zebrafish Danio rerio. Thesis (doctorate in Animal Science), 89 pp. São Paulo State University – UNESP: Botucatu, 2015.
Dammski, A.P., Muller, B.R., Gaya, C. & Regonato, D. (2011). Zebrafish – Manual of Breeding in Captivity. Federal University of Paraná, Curitiba, 107 pp.
De Bem, J.C., Fontanetti, C.S., Senhorini, J.A. & Parise-Maltempi, P.P. (2012). Effectiveness of estradiol valerate on sex inversion in Astyanax altiparanae (Characiformes, Characidae). Braz. Archives Biol. Technol. 55, 283–90.
Desai, K., Spikings, E. & Zhang, T. (2011). Effect of chilling on sox2, sox3 and sox19a gene expression in zebrafish (Danio rerio) embryos. Cryobiology 2, 96103.
Desai, K., Spikings, E. & Zhang, T. (2015). Short-term chilled storage of zebrafish (Danio rerio) embryos in cryoprotectant as an alternative to cryopreservation. Fish Haus 12, 11115.
Digmayer, M. (2010). Viability of pacu embryos, Piaractus mesopotamicus (Holmberg, 1887), submitted to −8°C and different concentrations of cryoprotectants. Dissertation (Master's Degree in Animal Science), 69 pp. Maringá State University – UEM: Maringá.
Elonen, G.E., Spehar, R.L., Holcombe, G.W., Johnson, R.D., Fernandez, J.D., Erickson, R.J., Tietze, J.E. & Cook, P.M. (1998). Comparative toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin to seven freshwater fish species during early life stage development. Environ. Toxicol. Chem. 17, 472.
Fernandes, K.A. (2016). Acute effects and morphological changes in larvae of Astyanax altiparanae (Garutti & Britski, 2000) (Teleostei, Characidae) exposed to extracts of cyanobacteria producing and not producing microcystin-LR. Dissertation (Master in Environment and Water Resources) 52 pp. Federal University of Itajubá – UNIFEI: Itajubá.
Ferraz, E.M. & Cerqueira, V.R. (2010). Influence of temperature on gonadal maturation of sea bass, Centropomus undecimalis. Bol. Inst. Pesca 36, 7383.
Fornari, D.C., Ribeiro, R.P., Streit, D.P. Jr., Vargas, L., Godoy, L.C., Oliveira, D., Digmayer, M., Galo, J.M. & Neves, P.R. (2012). Increasing storage capability of pacu (Piaractus mesopotamicus) embryos by chilling: Development of a useful methodology for hatcheries management. Cryo-Letters 33, 126–34.
Godoy, L.C., Zampolla, T., Streit, D. Jr., Bos-Mikich, A. & Zhang, T. (2012). Vitrification of zebrafish (Danio rerio) ovarian follicles. Cryobiology 65, 3, 344.
Goltermann, H.L., Clymo, R.S. & Ohnstad, M.A.M. (1978). Methods for Physical and Chemical Analysis of Freshwaters. London: Blackwell Science Publication, IBP Handbook Number 8, 214 pp.
Kimmel, C.B., Ballard, W.W., Kimmel, S.R., Ullmann, B. & Schilling, T.F. (1995). Stages of embryonic development of the zebrafísh. Dev. Dynam. 203, 253310.
Lattman, E.E., Fiebig, K.M. & Dill, K.A. (1994). Modelling compact denatured states of proteins. Biochemistry 33, 6158–66.
Leatherland, J.F. & Woo, P.T. (eds). (1998) Fish Diseases Non-Infectious Disorders. CABI, 2, 379 pp.
Leme dos Santos, H.S. & Azoubel, R. (1996). Comparative Embryology. FUNEP, Jaboticabal, 189 pp.
Little, E.E., Fairchild, J.F. & De Lonay, A.J. (1993). Behavioral methods for assessing impacts of contaminants on early life stage fishes. Am. Fish. Soc. Symp. 14, 6776.
Lopes, T.S., Strei, D. Jr., Ribeiro, R.P. & Romagosa, E. (2014). Post-cooling damage to Piaractus mesopotamicus embryos at different stages of development. Atas de Saúde Amb. 2, 111.
Miliorini, A.B., Murgas, L.D.S., Viveiros, A.T.M., Franciscatto, R.T., Silva, M.O.B. & Maria, A.N. (2002). Cooling of pacu semen (Piaractus mesopotamicus) at 4°C using different concentrations of dimethylsulfoxide. Rev. Bras. Reprod. Anim. 26, 209–11.
Morris, G.J. & Watson, P.F. (1984). Cold shock injury – a comprehensive bibliography. Cryo-Lett. 5, 352–72.
Nagamatsu, P.C. (2013). Effects of neurotoxic metals on Rhamdia quelen larvae exposed in early stages of development. Dissertation (Master's Degree in Cellular and Molecular Biology), 85 pp. Federal University of Paraná – UFPR: Curitiba.
Nakatani, K., Agostinho, A.A. & Baumgartner, G. (2001). Freshwater Fish Eggs and Larvae: Development and Identification Manual. Nupélia, Maringá, 359 pp.
Neves, P.R., Ribeiro, R.P., Streit, D.P. Jr., Natali, M.R., Fornari, D.C., Santos, A.I. & Godoy, L.C. (2014). Injuries in pacu embryos (Piaractus mesopotamicus) after freezing and thawing. Zygote 22, 2531.
Ninhaus-Silveira, A., Foresti, F., Azevedo, A. et al. (2008). Cryogenic preservation of embryos of Prochilodus lineatus (Valenciennes, 1836) (Characiforme; Prochilodontidae). Zygote 17, 4555.
Paes, M.C.F., Silva, R.C., Nascimento, N.F., Valentin, F.N., Senhorini, J.A. & Nakaghi, L.S.O. (2014). Hatching, survival and deformities of piracanjuba (Brycon orbignyanus) embryos subjected to different cooling protocols. Cryobiology 69, 451–6.
Pinder, A.C. & Gozlan, R.E. (2004). Early ontogeny of sunbleak. J. Fish Biol. 64, 762–75.
Prasad, T.K. (1996). Mechanisms of chilling-induced oxidative stress injury and tolerance in developing maize seedlings: changes in antioxidant system, oxidation of proteins and lipids, and protease activities. Plant J. 10, 1017–26.
Rall, W.F. (1993). Recent advances in the cryopreservation of salmonid fishes. In Genetic Conservation of Salmonid Fishes (Cloud, J.G. & Thorgaard, G.H., eds), Plenum, New York pp. 137–58.
Rauen, U., Polzar, B., Stephan, H., Mannherz, H.G. & De Groot, H. (1999). Could induced apoptosis in cultured hepatocytes and liver endothelial cells: mediation by reactive oxygen species. FASEB J. 13, 115–68.
Rawson, D. & Zhang, T. (2005). New approaches to the cryopreservation of fish oocytes and embryos. The role of biotechnology. Villa Gualino, Turin, Italy – 5–7 March, pp. 209–10.
Sakamoto, S. & Yone, Y. (1978). Effect of starvation on hematological characteristics, and the contents of chemical components and activities of enzymes in blood serum of red sea bream. J. Fac. Agric. Kyushu Univ. 23, 63–9.
Stevanato, D.J. (2016). Larval and post-larval ontogeny of Astyanax altiparanae (Garutti & Britski, 2000) in laboratory. Dissertation (Masters in Animal Science) 79 pp. Federal University of Paraná – UFPR: Curitiba.
Tajima, K. & Shimizu, N. (1973). Effect of sterol, alcohol and dimethyl-sulfoxide on sorghum seedling damaged by above-freezing low temperature. Proc. Crop. Sci. Soc. Jpn. 42, 220–6.
Tolosa, E.M.C., Rodrigues, C.J., Behmer, O.A. & Freitas-Neto, A.G. (2003). Manual of Techniques for Normal and Pathological Histology. Manole, Barueri, 341 pp.
Walker, M. K. & Peterson, R.E. (1991). Potencies of polychlorinated dibenzo-p-dioxin, dibenzofuran, and biphenyl congeners, relative to 2,3,7,8-tetrachlorodibenzo-p-dioxin, for producing early life stage mortality in rainbow trout (Oncorhynchus mykiss). Aquat. Toxicol. 21, 219–37.
Wood, K.A. & Youle, R.J. (1995). The role of free radicals and p53 in neuron apoptosis in vivo. J. Neurosci. 15, 8, 5851–7.
Zanardi, M.F., Dias-Koberstein, T.C.R, Dos Santos, M.A. & Malheiros, E.B. (2011). Productive performance and sexual reversal in tilapia in two hormonal methods. Vet. Zootec. 18, 2436.

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