Viability of dourado embryos cooled in different cryoprotectant solutions

Fornari, Darci Carlos; Júnior, Danilo Pedro Streit; Galo, Juliana Minardi; Guerreiro, Luis Ricardo Jayme; Povh, Jayme Aparecido; Barrero, Nelson Mauricio Lopera; Oliveira, Diego de; Ribeiro, Ricardo Pereira

doi:10.1590/s0100-204x2013000800057

Cited by 4 publications

(4 citation statements)

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“…The addition of sucrose may promote better cell dehydration and effectively decrease intracellular ice formation in a shorter period of time 26 . Moreover, the addition of sucrose could affect post-thaw viability 27 28 . By interacting with the polar heads of the phospholipids, sucrose may have stabilised the membrane when ice crystals grew 29 .…”

Section: Discussionmentioning

confidence: 99%

Cryopreservation of the gorgonian endosymbiont Symbiodinium

Chong

Tsai

Wang

et al. 2016

Sci Rep

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The study focused on finding a suitable cryoprotectant (CPA) and an optimum freezing protocol for the cryopreservation of the endosymbiotic dinoflagellates (Symbiodinium, clade G) of Junceella fragilis wherein the success of experiments is crucial to both scientific and ecology studies. A two-step freezing technique was developed. The viability of the thawed dinoflagellates was assayed using the adenosine triphosphate (ATP) bioassay for the first time and was further confirmed through the culturing of dinoflagellates in vitro. The results suggested that 30 min was the most suitable holding time for the dinoflagellates, and the samples produced highest viability when suspended at 5 cm from the surface of LN2. Results also showed that 1 M methanol with 0.4 M sucrose was the most effective CPA, yielding the highest viability (56.93%). Although cell densities of both cryopreserved and control group suffered an initial decline of culture, the cell densities were maintained throughout the remaining duration. In the present study, the cryopreservation of clade G endosymbiont algae was studied for the first time and the method described here could be applied for future studies on symbiotic algae cryopreservation.

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Section: Discussionmentioning

confidence: 99%

Cryopreservation of the gorgonian endosymbiont Symbiodinium

Chong

Tsai

Wang

et al. 2016

Sci Rep

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“…Pesquisa realizada por Zhang e Rawson (1995) avaliando crioprotetores para embriões de Danio rerio, também sugeriram o uso de metanol associado com sacarose (0,1M). Outro trabalho que também corrobora com os resultados do presente experimento foi realizado com embriões de espécies sul americana por Fornari et al (2013) que observaram eficiência do metanol associado a sacarose no resfriamento de embriões de Salminus brasiliensis.…”

Section: Resultsunclassified

Crioprotetores para o resfriamento de embriões de piracanjuba Brycon orbignyanus (Valenciennes, 1850)

Fornari

Streit

Tonissi³

et al. 2014

Sem. Ci. Agr.

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ResumoO objetivo deste trabalho foi avaliar a eficiência dos crioprotetores para o resfriamento de embriões de B. orbignyanus. Foram selecionados 1.400 embriões viáveis, na fase de pós-gastrulação e submetidos ao resfriamento a -8ºC com diferentes combinações de crioprotetores permeáveis (Metanol e dimetilsulfóxido -DMSO 9% ou associados 4,5%) combinados à crioprotetores não permeáveis (Sacarose e Glicose, 17%). Para cada tratamento, 200 embriões foram alocados em tubos de vidro "vacutainer" e submetidos ao processo de resfriamento. Após o tempo proposto, os embriões foram retirados e transferidos para incubadoras de quatro litros até a eclosão das larvas. Os resultados com as soluções contendo glicose no resfriamento foram ineficiente, apenas 27% dos embriões sobreviveram no tratamento. Os tratamentos com associação do DMSO não houve sobrevivência. Para o resfriamento de embriões de B. orbignyanus exposto a -8°C por um período de seis horas deve-se utilizar solução crioprotetora contendo sacarose 17% associada ao metanol 9%. Palavras-chave: Biotecnologia, criopreservação, crioprotetor, metanol, sacarose AbstractThe aim of this study was to evaluate cryoprotectants for cooling embryos of B. orbignyanus. 1400 embryos were selected, in the post-gastrulation and subjected to cooling to -8°C with different combinations of intracellular cryoprotectants (methanol and 9% DMSO or associated 4.5%) combined with extracellular cryoprotectants (sucrose and glucose, 17%) diluted in distilled water to complete 100%. For each treatment, 200 embryos were placed in glass tubes and submitted to the cooling process. After the proposed time, the embryos were removed and transferred to four-liter incubation to hatching of the larvae. The results obtained from solutions containing glucose were inefficient. The treatments with association of DMSO showed no satisfactory results. For cooling of B. orbignyanus embryos at -8°C it is suggested to use a cryoprotectant solution containing 17% sucrose associated with 9% methanol.

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“…We chilled Piaractus mesopotamicus embryos using a chilling solution composed by 0.5 M (17.1%) sucrose and 2.81 M (9%) methanol for six hours at-8 °C, which resulted in a hatching rate of 69.2% [21]. The same protocol was used for other South American fish embryos, achieving good results as for Brycon orbygnianus with hatching rate above 62% [34], Rhinelepis aspera upper 50% [35] (Figure 1), for Salminus brasiliensis with hatching rate of 67.06 % [36], and for Colossoma macropomum achieving a hatching rate of 77.6% [36].…”

Section: Chillingmentioning

confidence: 99%

Cryopreservation of Embryos and Oocytes of South American Fish Species

Streit¹,

Godoy²,

Pereira-Ribeiro³

et al. 2014

Recent Advances in Cryopreservation

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Additional information is available at the end of the chapter http://dx.doi.org/10.5772/58703 . IntroductionThe importance of animal genetic resources for wildlife maintenance as well as farming production has become more and more evident in recent years. Fish stocks are globally threatened mainly due to overfishing and environmental pollution. Cryopreservation of aquatic germplasm brings the possibility of preserving the genome of endangered species, increasing the representation of genetically valuable animals for farming purposes and avoiding genetic losses through diseases and catastrophes.In fish, successful cryopreservation of semen from many species has been well documented and cryopreserved semen has been used for reproduction of many wild and farmed species. "ttempts to cryopreserve fish embryos have been conducted over the past three decades, nevertheless successful cryopreservation protocol for long-term storage still remains elusive.In this chapter we cover the cryobiology applications on assisted reproduction for fish farming, focused on embryo and oocyte cryopreservation. Our research group has been working in this area for more than years and the chapter shows the main results that we have achieved from researches on embryos cryopreservation. The barriers that have been identified as a hamper for successful cryopreservation and the sensitiveness of tropical fish embryos to low temperature exposure are also detailed here.Recently, researches have reported that the use of oocytes may offer some advantages when compared to fish embryos, improving the chances of a successful cryopreservation. On this point, the chapter presents the most recent achievements in oocytes cryopreservation from South "merican fish, new trends and future works on this area.© 2014 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. . Brazilian aquaculture growth and researches in cryobiology areaThe history of "razilian aquaculture dates back to 's when the first scientific studies related to fish farming were carried out by Rodolpho von Ihering [ ], however, the farming activities had bit higher impulses only from the 's by creation of research center facilities like CEPT" Center for Research and Training in "quaculture and the insertion of academic activities related to aquaculture at universities. From the mid 's the entry of some private companies boosted the sector, and from the 's aquaculture started to represent an important activity of the "razilian agribusiness.Fish farming is the sector of the "razilian animal production industry which has displayed the highest growth in recent years, reaching , t in , representing an increase of . % compared to its production in [ ]. From the total fish farmed in , about % , t of the production was from South "merican species [ ].The growth in fish...

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Viability of dourado embryos cooled in different cryoprotectant solutions

Cited by 4 publications

References 12 publications

Cryopreservation of the gorgonian endosymbiont Symbiodinium

Cryopreservation of the gorgonian endosymbiont Symbiodinium

Crioprotetores para o resfriamento de embriões de piracanjuba Brycon orbignyanus (Valenciennes, 1850)

Cryopreservation of Embryos and Oocytes of South American Fish Species

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