Cryopreservation: Extending the viability of biological material from sea urchin (Echinometra lucunter) in ecotoxicity tests

Ribeiro, Raphaela Cantarino; Veronez, Alexandra Caroline da Silva; Tovar, Thaís Tristão; Adams, Serean L.; Bartolomeu, Dayse Aline Ferreira Silva; Perônico, Clayton; Furley, Tatiana Heid

doi:10.1016/j.cryobiol.2017.10.002

Cited by 19 publications

(12 citation statements)

References 25 publications

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“…In general, very few cold treatments are applied to marine ecotoxicological studies, and only limited to cryopreservation (Paredes and Bellas 2009;Cantarino Ribeiro et al 2018).…”

Section: Discussionmentioning

confidence: 99%

“…Cryopreservation of marine invertebrates has been proposed for several development stages and cell types (i.e., sperm, oocytes, embryos and larvae) from a wide number of different species (Guo and Weng 2020), with a high commercial and ecological value. Specific cryopreservation protocols have been developed for marine vertebrates and invertebrates (Paredes 2015;2019;Asturiano et al 2017), mainly on species of global economic importance, including fish, sea urchins, and mollusks used in aquaculture (Campos et al 2021;Cantarino Ribeiro et al 2018). As to crustaceans, few studies have been proposed for larvae cryopreservation, for example for penaeid prawn larvae (Arun and Subramoniam 1997;Subramoniam and Arun 1999) and barnacles larvae, including Balanus improvisus and A. amphitrite (Gakhova et al 1990;Khin-Maung-Oo et al 1998).…”

Section: Introductionmentioning

confidence: 99%

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Cold storage effects on lethal and sublethal responses of Amphibalanus amphitrite Nauplii

et al. 2022

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Bioassays are extensively used in ecotoxicology and there is a constant need for even more sensitive, reliable and easy to rear and obtain model organisms. Larvae of the crustacean Amphibalanus amphitrite are a good ecotoxicological model, for their high sensitivity to a wide range of toxicants and emerging contaminants. A standardized protocol for this toxicity bioassay has been recently proposed. Nevertheless, a limit of this model organism is the lack of resting stages and the need to use larvae immediately after their release from adults, thus increasing laboratory efforts related to the maintenance of adults. The aim of this work is to verify if short-term cold storage of A. amphitrite larvae prior to use in ecotoxicological tests may affect the ecotoxicological responses of these organisms. Three end-points (mortality, immobilization and swimming speed alteration) were measured on nauplii after storing them at 4 ± 1 °C for different times (24, 72 and 120 h) before bioassay set-up. Bioassays were set up using: (i) clean filtered natural sea water (0.22 µm FNSW), (ii) a reference toxicant (Cadmium Nitrate) and (iii) an environmental matrix (sediment elutriate). Results show that mortality, differently from the other two endpoints, was not affected by cold-storage. Even after 5 days of larvae storage at 4 ± 1 °C before bioassay set up, mortality data were comparable to those obtained for non-cold-stored organisms. Moreover, larval sensitivity to the reference toxicant and sediment elutriate did not change. Regarding the other two end points, low cadmium concentrations significantly changed immobility and swimming activity in cold-stored nauplii compared to larvae used immediately after larval release. In conclusion, short-term cold storage of A. amphitrite nauplii before bioassay set up is an appropriate procedure in ecotoxicological testing if mortality is the endpoint to be considered for final evaluation.

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“…In general, very few cold treatments are applied to marine ecotoxicological studies, and only limited to cryopreservation (Paredes and Bellas 2009;Cantarino Ribeiro et al 2018).…”

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Cold storage effects on lethal and sublethal responses of Amphibalanus amphitrite Nauplii

et al. 2022

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“…This increased sensitivity may be because cryopreserved organisms are going through a recovery process after thawing, and might be more sensitive to additional stress, such as toxicant exposure. Ribeiro et al (2018) developed a cryopreservation protocol for Echinometra lucunter sperm and they are studying the cryopreservation of embryos for water quality assessment. There is a cryopreservation protocol described for P. lividus sperm (Fabbrocini et al, 2014) that yields good motility.…”

Section: Cryopreserved Echinoidsmentioning

confidence: 99%

The Use of Cryopreserved Biological Material for Water Quality Assessment

Paredes

Bellas

2019

Front. Mar. Sci.

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The stated aim of this perspective article is to present new developments and discuss future directions on the applications of cryopreserved organisms to marine water quality assessment. To facilitate this, the authors provide a background of essential knowledge of cryopreservation when applied to ecotoxicology, as well as, practical examples available in literature. An integrated approach with combined monitoring of chemical status plus measurements of biological effects has been recommended extensively by international institutions for the assessment of marine pollution. Among the available techniques, bioassays have been considered as sufficiently robust to be incorporated in marine pollution monitoring programs. However, the routine application of bioassays has also allowed the identification of one of the factors that limits a more extensive use of such biological methods: the availability of biological material throughout the year, regardless of natural spawning periods. A solution to this limitation is the application of cryopreservation techniques. Cryopreservation may, for instance, provide access to stable quality biological material when test species are out of the reproductive season, without the need for maintaining and conditioning organisms in the laboratory. It also guarantees access to a large variety of species that might not be available at the same time of the year and, on top of that, cryopreservation provides opportunities to laboratories that might not have the facilities to keep all these organisms in culture.

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“…The history of cryopreservation of sea urchin sperm began in 1973 (9), and at present, sperm of about 15 species have been cryopreserved as recently reviewed (11,26-28,31). Cryopreservation of sperm of all animals involves dilution of concentrated semen into an extender solution and then mixing this cell suspension with a cryoprotectant such as dimethyl sulfoxide (DMSO).…”

Section: Introductionmentioning

confidence: 99%

Cold Storage and Cryopreservation Methods For Spermatozoa of the sea urchinLytechinus pictus

Vacquier,

Hamdoun

2023

Preprint

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Sea urchins have contributed greatly to knowledge of fertilization, embryogenesis and cell biology. However, until now, they have not been a genetic model organism because of the long generation times of commonly used species, and lack of tools for husbandry and genetic manipulation. We recently establishedLytechinus pictus, as a multigenerational sea urchin model, because of its relatively short generation time of 4-6 months and ease of laboratory culture. To take full advantage of this new multigenerational species, methods are needed to biobank and share mutantL. pictussperm. Here, we describe a new extender based on sperm ion physiology before spawning of sperm into seawater. This extender maintains sperm capable of fertilization for at least 5-10 weeks when stored at 0 °C. We use the extender, and the cryoprotectant dimethyl sulfoxide (DMSO), to cryopreserve sperm of bothL. pictus, and the widely used sea urchin,Strongylocentrotus purpuratus. The simple methods we describe work well for both species, achieving > 90% development and producing larvae that successfully undergo metamorphosis to juvenile sea urchins. Sperm of these two species can be frozen and thawed at least twice and still give rise to larvae that undergo metamorphosis.Main PointsSperm can maintain fertilizing capacityex vivofor 5-10 weeks when stored at 0°C.When freezing in liquid nitrogen no stepwise addition of cryoprotectant, or stepwise drop in temperature are required.A standard fertilization assay is presented to score cleavage stage sea urchin embryos produced by cryopreserved sperm.Sperm frozen and thawed more than once can produce larvae.

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Cryopreservation: Extending the viability of biological material from sea urchin (Echinometra lucunter) in ecotoxicity tests

Cited by 19 publications

References 25 publications

Cold storage effects on lethal and sublethal responses of Amphibalanus amphitrite Nauplii

Cold storage effects on lethal and sublethal responses of Amphibalanus amphitrite Nauplii

The Use of Cryopreserved Biological Material for Water Quality Assessment

Cold Storage and Cryopreservation Methods For Spermatozoa of the sea urchinLytechinus pictus

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