• Journal of Marine Life Science
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• v.5 no.2
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• pp.92-98
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• 2020

Heat shock proteins (HSPs) are highly conserved cellular proteins that contribute to adaptive responses of organisms to a variety of stressors. In response to stressors, cellular levels of HSPs are increased and play critical roles in protein stability, folding and molecular trafficking. The mRNA expression pattern of two well-known heat shock protein transcripts, HSP70 and HSP90 were studied in two tissues of nerve ganglia, cerebral ganglion and pleuropedal ganglion of Pacific abalone (Haliotis discus hannai). It was observed that both HSP70 and HSP90 transcripts were upregulated under heat stress in both ganglion tissues. Expression level of HSP70 was found higher than HSP90 in both ganglia whereas cerebral ganglion showed higher expression than pleuropedal ganglion. The HSP70 and HSP90 showed higher expression at Day-1 after exposed to heat stress, later decreased at Day-3 and Day-7 onwards. The present result suggested that HSP70 and HSP90 synthesize in nerve ganglion tissues and may provide efficient protection from stress.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.3
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• pp.223-231
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• 2020

Karyotype analysis is a major work in the process of triploid abalone production for the purpose of productivity and quality improvement. However, the metaphase spreads for karyotype analysis have been prepared just from the larvae at trochophore stage, which has restricted the spectrum of sample correction inhibiting more efficient analysis. Here, we investigated the feasibility of preparing metaphase spreads from the larvae at veliger stage that is the next developmental stage of trochophore. For this, diploid and triploid larvae at trochophore and veliger stages from Pacific abalone (Haliotis discus hannai) were subjected to metaphase spread preparation and its efficiencies were measured and compared each other. As the results, although the efficiencies of metaphase spread preparation were significantly lower in the larvae at veliger stage compared to the ones at trochophore stage regardless of ploidy status, we found that the preparation of metaphase spreads, which showed the clear chromosomal images containing the normal number of chromosomes, was possible from the veliger stage larvae. On the other hands, all larvae used in this study regardless of developmental stage and ploidy did not show colchicine sensitivity. Moreover, no significant difference was observed in cell cycle distribution of the cells comprising larvae between two developmental stages regardless of ploidy status. These suggested that the details of protocol to prepare metaphase spreads from abalone larvae should be optimized depending on its developmental stages. Taken together, we demonstrated the feasibility of preparing metaphase spreads from H. discus hannai veliger stage larvae for karyotype analysis.

• Journal of Animal Reproduction and Biotechnology
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• v.34 no.4
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• pp.280-291
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• 2019

Potential utility of 14 candidate housekeeping genes as normalization reference for RT-qPCR analysis with developmental samples (fertilized eggs to late veliger larvae) in Pacific abalone Haliotis discus hannai was evaluated using four different statistical algorithms (geNorm, NormFinder, BestKeeper and comparative ΔCT method). Different algorithms identified different genes as the best candidates, and geometric mean-based final ranking from the most to the least stable expression was as follow: RPL5, RPL4, RPS18, RPL8, RPL7, UBE2, RPL7A, GAPDH, RPL36, PPIB, EF1A, ACTB and B-TU. The findings were further validated via relative quantification of metallothionein (MT) transcripts using the stable and unstable reference genes, and expression levels of MT were greatly influenced according to the choice of reference genes. In overall, our data suggest that RPL5 and RPS18, either singly or in combination, are appropriate for normalizing gene expression in developmental samples of this abalone species, whereas ACTB, B-TU and EF1A are less stable and not recommended. In addition, our findings propose that standard deviations in geometric ranking as well as geometric mean itself should also be taken into account for the final selection of reference gene(s). This study could be a useful basis to facilitate the generation of accurate and reliable RT-qPCR data with developmental samples in this abalone species.

• The Korean Journal of Malacology
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• v.32 no.4
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• pp.249-254
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• 2016

The purpose of this study is to estimate genetic parameter and growth traits by sex of Pacific abalone, Haliotis discus hannai. The abalone 10 family produced using 1:1 mating system of male and female for analyses of sex ratio and growth traits (shell length, shell width, and total weight) by sex. Overall mean in phenotypic traits at 30-month-old showed 85.01 mm of shell length, 57.49 mm of shell width and 73.34 g of total weight respectively. The sex ratio (female : male) was 1:0.93 (n = 191:177). The values of growth traits by sex showed significant difference (P < 0.05). The each value of female growth traits were shown to be higher than the values of male growth traits. The heritability of growth traits by sex were estimated that the heritability of female growth traits are higher than male that. The results suggest a possibility of improving the growth of cultured abalone using selection breeding by sex.

• The Korean Journal of Malacology
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• v.32 no.4
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• pp.241-247
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• 2016

Macrophage Migration Inhibitory Factor (MIF) are well-defined role as unique cytokine and critical mediator in acute and chronic inflammatory diseases, autoimmune diseases. In this study, we isolated and characterized a full-length of MIF cDNA from the abalone (Haliotis discus hannai). The full-length cDNA of abMIF was of 1264 bp, consisting of a 5'-terminal UTR of 143 bp, an open reading frame of 360 bp and a 3-terminal UTR of 761 bp. The abalone MIF cDNA encodes a 119-amino acid polypeptide with a calculated molecular mass of 13.4 kDa and isoelectric point of 9.07. Multiple alignments and phylogenetic analysis with the deduced abalone MIF protein and showed strong homology with disk abalone (Haliotis discusdiscus). The deduced amino acid sequence of abMIF exhibited homology with other reported MIFs, such as 80%, with that of other disk abalone H. discus discus MIF gene. Quantitative real-time PCR (qRT-PCR) analysis indicated that abMIF was highly expression observed in hapatopacreas, intestine, foot, and gonad of normal conditioned abalone. Even though AbMIF mRNA level in hemocytes was low under the normal condition, it was sharply up-regulated and reached the maximum at 6 h post-infection with Vibrio parahaemolyticus, and then decreased at 24 h post-infection. This result indicates that abMIF plays an important role in responding in the innate immune system.

• The Korean Journal of Malacology
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• v.28 no.4
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• pp.343-347
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• 2012

The growth rates of offspring of selected abalone (SA), Haliotis discus hannai, population for rapid growth were compared to those of offsping of the control abalone (CA) at bred under the same condition. The growth traits (shell length, shell breadth and total weight) of SA were significantly faster than those of CA at this experiment. At the 45th day and 90th day, the growth rates in shell length of SA were estimated 31.7% and 17.8% faster than those of CA, total weight of SA were estimated 43.7% and 25.6%, respectively. Moreover, the relative growth rate between shell length and total weight of SA showed significant differences higher than one in CA at 90th day. The results suggest a possibility of improving the growth rate of cultured abalone using selection techniques.

• Proceedings of the Zoological Society Korea Conference
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• 2003.08a
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• pp.209.4-209
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• 2003

No Abstract, See Full Text

• Korean Journal of Environmental Biology
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• v.35 no.4
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• pp.670-676
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• 2017

Recently, mass mortality of the young abalone Haliotis discus hannai has occurred in commercial seed production farms in Korea. The mortality rate was above 50% of the total cultured organisms in the farm, and the shell length of the moribund organisms was about 3cm. The mortal phenomenon was that the young abalones were weakly scattered on the bottom of the pond from the attachment matrix, or that they could not be moved back to their normal positions. The diseased farmed Pacific abalone had abdominal edema. From the edema in the moribund individuals, three bacterial strains were isolated and all the strains were identified as Vibrio harveyi. These strains were compared with thirty six strains isolated from the fish. The results was that the Vibrio harveyi from the fish were sorted into genogroup A or B; however, the three strains of the diseased farmed Pacific abalone were sorted into genogroup A and the new genogroup C. The identical mortality and pathological symptoms of the naturally infected organisms were reproduced by artificial infection with WA AG-1 and WA CS-5 strains. The $LD_{50}$ of WA AG-1 and WA CS-5 were each $1.0{\times}10^3cfu\;animal^{-1}$ and $1.7{\times}10^4cfu\;animal^{-1}$.

• Fisheries and Aquatic Sciences
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• v.18 no.1
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• pp.81-88
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• 2015

We investigated factors affecting primary cultures of Pacific abalone Haliotis discus hannai ovary-dissociated cells to identify general aspects of their early-phase culture. Ninety-seven cell populations derived from 30 individuals were cultured in different media with varying compositions of medium supplements, and initial attachment, subculture, and survival for ${\geq}10$ weeks were assessed according to medium composition and individual. We also examined the time required for subculture and the rate of cell death according to both culturing period and passage number within 10 weeks. A lack of fetal bovine serum (FBS) and hemolymph significantly inhibited the growth of cultured cells, while we detected no significant effect of medium composition on initial cell attachment. Through data reallocation, with the omission of data from cell populations cultured in FBS-free and hemolymph-free media, we showed that growth inhibition was also affected by individual differences among the abalones used. During the culture, we observed four different types of cell morphology. Moreover, considerable time was required for subculture-18.4 and 19.5 days for first and second subcultures, respectively-and cell death did not occur within 30 days or for passage 0. Our results will provide valuable information for developing universal cell culturing guidelines in abalone species and suggest the feasibility of culturing abalone ovary-dissociated cells.

• Animal cells and systems
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• v.14 no.4
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• pp.305-313
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• 2010

Pacific abalone Haliotis discus discus is an important fisheries resource in Jeju, Korea. For basic information about its current genetic status in relation to stock enhancement, the level and distribution of genetic variation between wild and released stocks of Pacific abalone in Jeju were examined at nine cross-species microsatellite markers including the use of two novel primers. High levels of polymorphism were observed between the two populations. A total of 146 different alleles were found at all loci, with some alleles being unique. The allelic variability ranged from five to 27 in the wild population and from four to 16 in the released sample. The average observed and expected heterozygosities were estimated to be 0.74 and 0.84 in the wild sample and 0.70 and 0.78 in the released sample, respectively. Although a considerable loss of rare alleles was observed in the released sample, no statistically significant reductions were found in heterozygosity or allelic diversity in the released sample compared to the wild population. Low but significant genetic differentiation was found between the wild and released populations. These results suggest that the intensive breeding practices for stock enhancement may have resulted in a further decrease in genetic diversity, and that the cross-species microsatellite markers used in this study represent a potentially efficient means for further genetic studies, providing beneficial information for the protection and management of H. discus discus.