Immobilisation of living coral embryos and larvae

Randall, Carly J.; Giuliano, Christine; Mead, David A.; Heyward, Andrew; Negri, Andrew P.

doi:10.1038/s41598-019-51072-5

Cited by 3 publications

(3 citation statements)

References 19 publications

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“…Large-particle flow cytometry has also been applied to the screening and sorting of microencapsulated particles, cells, and organisms, such as fungal spores encapsulated in calcium alginate beads 18 . A recent investigation that immobilised several species of coral propagules in an agarose gel found that survival of immobilised larvae was high, and that the encapsulation process did not significantly impede larval settlement after immobilisation 42 . A work flow could be developed whereby the output from automated encapsulation feeds directly into a large-particle flow cytometer to measure and sort individually-encapsulated particles.…”

Section: Discussionmentioning

confidence: 98%

Rapid counting and spectral sorting of live coral larvae using large-particle flow cytometry

Randall

Speaks

Lager³

et al. 2020

Sci Rep

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Research with coral embryos and larvae often requires laborious manual counting and sorting of individual specimens, usually via microscopy. Because many coral species spawn only once per year during a narrow temporal window, sample processing is a time-limiting step for research on the early life-history stages of corals. Flow cytometry, an automated technique for measuring and sorting particles, cells, and cell-clusters, is a potential solution to this bottleneck. Yet most flow cytometers do not accommodate live organisms of the size of most coral embryos (> 250 µm), and sample processing is often destructive. Here we tested the ability of a large-particle flow cytometer with a gentle pneumatic sorting mechanism to process and spectrally sort live and preserved Montipora capitata coral embryos and larvae. Average survival rates of mechanically-sorted larvae were over 90% and were comparable to those achieved by careful hand-sorting. Preserved eggs and embryos remained intact throughout the sorting process and were successfully sorted based on real-time size and fluorescence detection. In-line bright-field microscopy images were captured for each sample object as it passed through the flow-cell, enabling the identification of early-stage embryos (2-cell to morula stage). Samples were counted and sorted at an average rate of 4 s larva −1 and as high as 0.2 s larva −1 for high-density samples. Results presented here suggest that large-particle flow cytometry has the potential to significantly increase efficiency and accuracy of data collection and sample processing during time-limited coral spawning events, facilitating larger-scale and higher-replication studies with an expanded number of species. Coral populations have suffered widespread losses over the last half century as a result of local, regional, and global pressures driven by human activities 1-4. Research on the early life-history stages of corals is foundational to our understanding of coral bioecology and is an increasingly active area of research as we enter a new era of restoration and adaptation science 5. Yet, research on the early stages of broadcast spawning corals is severely restricted by spawning events that only occur on a few nights of the year for a given species and location 6,7. These arduous periods often are defined by long working hours, demands on equipment and resources shared across research groups, and hence missed research opportunities. Much research relies on the painstaking process of counting and hand-sorting individual sub-millimetre diameter oocytes, embryos and larvae into well plates, test tubes, or petri dishes 8. Furthermore, rapid rates of cleavage 9 leave little time to work with early developmental stages. Investigations that employ CRISPR/Cas9, for example, require fertilized but not yet cleaving eggs 10 , offering as little as a few hours per year for research. A rapid and reliable method for sorting coral eggs, embryos and larvae, capturing population-level data, and sorting the samples based on condition would en...

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Section: Discussionmentioning

confidence: 98%

Rapid counting and spectral sorting of live coral larvae using large-particle flow cytometry

Randall

Speaks

Lager³

et al. 2020

Sci Rep

Self Cite

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“…Coral planulae have been studied to understand the skeletogenesis in scleractinians (le Tissier, 1988) and the effects of factors such as pollution (Rinkevich and Loya, 1979). Studies have also tested new biotechnology for coral reef research and restoration (Randall et al, 2019).…”

Section: Discussionmentioning

confidence: 99%

First successful production of adult corals derived from cryopreserved larvae

Narida

Tsai

Hsieh³

et al. 2023

Front. Mar. Sci.

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Coral reefs worldwide are declining due to increasing concentrations of greenhouse gases, which, combined with local anthropogenic pressure, are exacerbating unprecedented mass coral bleaching. For corals to survive, restoring coral reefs through cryopreservation is crucial. The aim of this study was to vitrify and laser-warm Stylophora pistillata planulae to allow for feasible settlement, post-settlement survival, and the production of adult corals. The no-observed-effect concentrations were used to determine the best cryoprotective agents for S. pistillata. The larvae were then subjected to cooling and nanolaser warming (300 V, 10 ms pulse width, 2 mm beam diameter) by using two vitrification solutions (VSs; VS1: 2 M dimethyl sulfoxide and 1 M ethylene glycol [EG]; VS2: 2 M propylene glycol and 1 M EG) and gold nanoparticles. The results revealed that VS1-treated larvae had a higher vitrification rate (65%), swimming rate (23.1%), settlement rate (11.54%), and post settlement survival rate (11.54%) than those treated with VS2. Seasonal variations also affected the cryopreservation of the planulae; VS1 was more favorable for the planulae in spring than in fall. Although laser-warmed larvae developed slower morphologically than their controlled counterparts, the production of cryopreserved adult S. pistillata corals was achieved. The proposed technique can improve the cryopreservation of corals and advance efforts to protect endangered coral species.

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“…For example, mechanisms that improve coral recruitment (i.e., survivorship rates of recruitment) could be an important research frontier to enhance coral population recovery after disturbance. Examples include innovations in new materials (e.g., hydrogels) to protect corals in vulnerable early life stages and increase survivorship of coral recruits (Randall et al, 2019), the incorporation of crustose coralline algae and biofilms in restoration projects (Heyward & Negri, 1999), or the use of acoustic playback of a healthy reef to enhance coral settlement in degraded sites (Gordon et al, 2019; Lillis et al, 2016).…”

Section: Future Directionsmentioning

confidence: 99%

A roadmap to integrating resilience into the practice of coral reef restoration

Shaver

McLeod

Hein³

et al. 2022

Global Change Biology

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Recent warm temperatures driven by climate change have caused mass coral bleaching and mortality across the world, prompting managers, policymakers, and conservation practitioners to embrace restoration as a strategy to sustain coral reefs. Despite a proliferation of new coral reef restoration efforts globally and increasing scientific recognition and research on interventions aimed at supporting reef resilience to climate impacts, few restoration programs are currently incorporating climate change and resilience in project design. As climate change will continue to degrade coral reefs for decades to come, guidance is needed to support managers and restoration practitioners to conduct restoration that promotes resilience through enhanced coral reef recovery, resistance, and adaptation. Here, we address this critical implementation gap by providing recommendations that integrate resilience principles into restoration design and practice, including for project planning and design, coral selection, site selection, and broader ecosystem context. We also discuss future opportunities to improve restoration methods to support enhanced outcomes for coral reefs in response to climate change. As coral reefs are one of the most vulnerable ecosystems to climate change, interventions that enhance reef resilience will help to ensure restoration efforts have a greater chance of success in a warming world. They are also more likely to provide essential contributions to global targets to protect natural biodiversity and the human communities that rely on reefs.

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Immobilisation of living coral embryos and larvae

Cited by 3 publications

References 19 publications

Rapid counting and spectral sorting of live coral larvae using large-particle flow cytometry

Rapid counting and spectral sorting of live coral larvae using large-particle flow cytometry

First successful production of adult corals derived from cryopreserved larvae

A roadmap to integrating resilience into the practice of coral reef restoration

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