Selection of cryoprotectants based on their toxic effects on oyster gametes and embryos

Nascimento, Iracema Andrade; Leite, Maria Bernadete Neiva Lemos; Araujo, Milena Maria Sampaio de; Sansone, Giovanni; Pereira, Solange Andrade; Santo, Maristela do Espírito

doi:10.1016/j.cryobiol.2005.04.006

Cited by 32 publications

(11 citation statements)

References 8 publications

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“…Our results generally correspond with the observation of Nascimento et al. () on the larvae of Crassostrea rhizoprae who found that the EC15–24 h (effective concentration of cryoprotectant that causes abnormalities in 15% of the population in 24 h) was 12.92% DMSO in case of a 30‐min exposure. Our choice of veligers or D‐shaped larvae for the candidate larval stage for cryopreservation was further supported by the findings of Choi and Chang () who found that this larval stage displayed the highest post‐thaw survival (91%) in the pearl oyster ( Pinctada furcata martensii ).…”

Section: Discussionsupporting

confidence: 92%

“…Oyster larvae were found to respond to 30-min exposure to DMSO in a dose-dependent manner. Our results generally correspond with the observation of Nascimento et al (2005) on the larvae of Crassostrea rhizoprae who found that the EC15-24 h (effective concentration of cryoprotectant that causes abnormalities in 15% of the population in 24 h) was 12.92% DMSO in case of a 30-min exposure. Our choice of veligers or D-shaped larvae for the candidate larval stage for cryopreservation was further supported by the findings of Á .…”

Section: Discussionsupporting

confidence: 92%

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Cryopreservation of sperm and larvae of the European flat oyster (Ostrea edulis )

et al. 2012

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Summary Cryopreservation of sperm and larvae of the European flat oyster (Ostrea edulis) was carried out to assess their performance characteristics using methods commonly employed in fish spermatozoa and other oyster species. Sperm was cryopreserved in Calcium‐free Hanks’ balanced salt solution in the presence of cryoprotectants methanol and dimethyl‐sulfoxide (DMSO) at concentrations of 5 or 10%. The highest post‐thaw motility (progressive: 3 ± 6%, static: 8 ± 10%) was observed with 10% DMSO with a noted decrease in comparison to fresh sperm (progressive: 13 ± 6%, static: 63 ± 23%). Membrane integrity of spermatozoa was measured by live‐dead fluorescent staining. The highest percentage of membrane‐intact cells (60 ± 6%) was also observed when 10% DMSO was used. A 30‐min exposure of trochophorae and veliger larvae to 5, 10, 15 or 20% DMSO was carried out to test the toxicity of this cryoprotectant on early life stages, resulting in the highest survival of veligers (99 ± 2%) exposed to 10% DMSO. Cryopreservation of veligers was carried out in two steps [(i) at 13 cm above the level of liquid nitrogen for 10 min, (ii) 10 cm for 12 or 23 min] in the presence of 5 or 10% DMSO. The highest post‐thaw survival of larvae (59 ± 31%) was observed at 12 cm of the second step and 10% DMSO. However, none of the larvae survived for 24 h after thawing probably due to the inaccuracies of the freezing protocol, thus, further studies are needed to standardize cooling conditions.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionsupporting

confidence: 92%

Cryopreservation of sperm and larvae of the European flat oyster (Ostrea edulis )

et al. 2012

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“…Cryoprotectant can penetrate the cell and make it dehydrated, protecting cells against the formation of intracellular ice crystals and impeding the reordering of water molecules F I G U R E 3 Effects of cryoprotectants type (DMSO, MET, PG, EG) and concentration (5, 10, 20, 30%) on velocity parameters (VCL, VSL, VAP) of the noble Scallop Chlamys nobilis [Colour figure can be viewed at wileyonlinelibrary.com] around super cold nuclei (Leung, 1991). Cryoprotectants are toxic to cells, especially at high concentrations, thus, the optimal cryoprotectant concentration should be a balance between its cryoprotective and toxic effects (Elliott, Wang, & Fuller, 2017;Nascimento et al, 2005). Therefore, a prudent first step is to screen permeability and low toxicity cryoprotectants for a particular cryopreservation protocol.…”

Section: Discussionmentioning

confidence: 99%

Sperm cryopreservation of the noble Scallop Chlamys nobilis by a programmable freezing method: Effect of cryoprotectant

et al. 2019

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A study on Chlamys nobilis sperm cryopreservation by a programmable freezing method was conducted under laboratory condition. Four cryoprotectant agents (dimethyl sulfoxide [DMSO], methanol [MET], propanediol[PG] and ethylene glycol [EG]) and four concentrations (5%, 10%, 20% and 30%) were evaluated for their ability to retain sperm motility, movement characteristics and fertility. Results showed that cryopreserved sperm total motility produced by DMSO and MET at 5%, 10% and 20% were higher than other cryoprotectant treatment groups (CPA groups), as well as rapid sperm percentage. The curvilinear (VCL) and straight line (VSL) velocity produced by DMSO at 5% significantly higher than other CPA groups (p < 0.05), while no significant differences were found for average path (VAP) velocity. The lateral head displacement (ALH) in all CPA groups was similar and without significant difference (p > 0.05), as well as the beat‐cross frequency (BCF). A significant higher fertilization rate was produced in DMSO than that in MET at same concentration (p < 0.05), and no significant differences were found for differing concentrations of the same cryoprotectant (p > 0.05). Overall, 5%‐20% DMSO was more suitable for Chlamys nobilis sperm programmable cryopreservation when the calcium‐free Hanks’ balanced salt solution was used as the extender, and 10°C/min from 0°C to −80°C was used as freezing rate. The findings presented in this study will benefit conservation programs for Chlamys nobilis.

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“…() observed up to 70% of progressive sperm motility after thawing using lower concentrations of PG and dimethyl sulfoxide (Me 2 SO), although lower survival rates were observed. For the mangrove oyster Crassotrea rhizophorae , MetOH was more toxic than PG in the assessment of normal D larvae obtained from normal oocytes fertilized using exposed sperm (Nascimento, Leite, Araújo, Sansone, Pereira & Espírito‐Santo ; Sansone, Nascimento, Leite, Araújo, Pereira & Mariani ). Similar results were obtained on Crassostrea gigas : normal oocytes were not fertilized by sperm cryopreserved using MetOH, while sperm frozen using PG at 5% and 10% concentrations achieved fertilization rates higher than 20% (Tervit, Adams, Roberts, McGowan, Pugh, Smith & Janke ).…”

Section: Discussionmentioning

confidence: 99%

Sperm motility and fertilization performance ofNodipecten nodosus(L., 1758) exposed at two different cryoprotectants

et al. 2014

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Cryopreservation is a valuable tool for aquaculture as it provides continuous seed production, regardless of the spawning season of the brood stock. The selection of a suitable cryoprotectant with low toxicity and high water solubility is important to avoid membrane injuries and intracellular ice crystallization. This study has been aimed at the assessment of the toxic effects of two usually applied cryoprotectants, 1-2 propylene glycol (PG) and methanol (MetOH), on spermatozoa of the of lion-paw scallop Nodipecten nodosus, by evaluating the sperm motility and the development of D larvae after fertilization procedure. Sperm was exposed at room temperature (22°C) for 10, 20 and 30 min to different concentration ranges of two cryoprotectants. Regarding the sperm motility, PG5%, PG7%, MetOH4% and MetOH6% did not show differences compared to control (semen incubated in seawater) (P < 0.05). The development of D larvae was not affected by the exposition to PG5%, MetOH 4% and MetOH 6%. These results indicate the potential use of both cryoprotectants for cryopreservation procedures.

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Selection of cryoprotectants based on their toxic effects on oyster gametes and embryos

Cited by 32 publications

References 8 publications

Cryopreservation of sperm and larvae of the European flat oyster (Ostrea edulis )

Cryopreservation of sperm and larvae of the European flat oyster (Ostrea edulis )

Sperm cryopreservation of the noble Scallop Chlamys nobilis by a programmable freezing method: Effect of cryoprotectant

Sperm motility and fertilization performance ofNodipecten nodosus(L., 1758) exposed at two different cryoprotectants

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