Survival of
zebrafish, Brachydanio rerio (Hamilton-Buchanan), embryo after
immersion in methanol and exposure to ultrasound with implications to
cryopreservation
A.N. Bart, H.A. Kyaw-2003
Aquaculture
Research, 34(8): 609-615
Abstract:
This study examined the viability of
embryos after immersion in highly concentrated methanol solutions (40-60%)
and exposing embryos to ultrasound to enhance efficient transport of the
cryoprotectant. The exposure to ultrasound, methanol concentrations,
duration of treatment and the stages of embryonic development was found to
have measurable effects on embryo viability. The effect of ultrasound was
more evident at high voltage (>440 V) settings and at early
developmental stages (30 and 60% epiboly stage). Older embryos were more
resistant to cryoprotectant toxicity and ultrasound-induced mortality. The
high concentration of methanol (60%) was more toxic to embryos than the low
concentration (40%). When methanol treatment and ultrasound were applied
simultaneously the optimum concentration was found to be 45% methanol (45%
survival; P<0.05) in a 3 min treatment. Although there was no
significant difference between the 2 and 3 min treatments, embryos
treated for 4 min had a significantly lower survival rate (P<0.05).
These findings provide initial results to select the developmental stage of
the embryo, the concentration of methanol for the preparation of a
vitrification solution and duration of ultrasound treatment for
cryopreservation. Furthermore, it indicates the potential use of ultrasound
to enhance the transport of methanol intracellularly with minimum mortality
of the developing embryos.
(Aquaculture and Aquatic Resources
Management, Asian Institute of Technology, PO Box 4, Klong Luang,
Pathumthani 12120, Thailand. E-mail: bart@ait.ac.th)