• Animal cells and systems
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• v.6 no.1
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• pp.75-80
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• 2002

A study on the effects of bisphenol A (BPA) on sex differentiation and gonadal development in medaka, Oryzias latipes, was investigated by histological examination. The fish were exposed to aqueous solutions of BPA at nominal concentrations of 50, 100, and 200 $\mu\textrm{g}$/L from newly-hatched larvae stage to 70 d. The ovaries of female fish were composed of oocytes at the chromatin nucleolus and peri-nucleolus stages at 20 d after the exposure. The testes contained a number of spermatogonia and spermatocytes at 30 d. In the process of sex differentiation. gonadal development was not different in all experimental groups until 30 d after the exposure. At 70 d after the exposure, however, advanced development of oocytes in the ovary and inhibition of spermatogenesis in the testis were observed in the BPA-treated groups compared to the non-treated controls. More females than males were identified in the 50 and 100 $\mu\textrm{g}$/L BPA-treated groups, in comparison to the 200 $\mu\textrm{g}$/L BPA-treated group and non-treated controls. Medaka exposed to 200 $\mu\textrm{g}$/L BPA were bigger compared to other experimental groups. The present study suggests that BPA may lead to problems in either mating or sexual behavior due to the difference in growth and disparity of sexual maturation between male and female fish.

• Development and Reproduction
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• v.21 no.1
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• pp.19-33
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• 2017

Optimum concentrations of anesthetic clove oil and anesthetic lidocaine-HCl were determined for a species of adult marine medaka, Oryzias dancena, over a range of salinity conditions, and investigated in a transport simulation experiment by analyzing various water and physiological parameters. Research indicated that the higher the concentration of anesthetic at each salinity, the shorter the anesthesia time at each salinity. At each concentration, fish were anesthetized slower at water salinities over 10 ppt (P<0.05). Anesthesia time at 10 ppt was faster than any other salinity. In 10 ppt salinity, the dissolved oxygen (DO) concentrations and respiratory frequencies of the clove-oil-administered groups decreased until 48 hours (P<0.05), whereas the $NH_4{^+}$ and $CO_2$ concentrations increased until 48 hours (P<0.05). In same period, the DO, $NH_4{^+}$, and $CO_2$ concentrations and respiratory frequencies all decreased as the clove oil concentration increased (P<0.05). The trends in the DO, $NH_4{^+}$, and $CO_2$ concentrations and respiratory frequencies in the lidocaine-HCl-administered groups were similar to those in the clove-oil-administered groups. In conclusion, clove oil and lidocaine-HCl are effective anesthetics, improving the transportation of the marine medaka. The results from this study will contribute to safe laboratory handling of the marine medaka, which are commonly required by many research studies and experiments.

• Korean Journal of Environmental Biology
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• v.30 no.4
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• pp.314-318
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• 2012

The aims of the present study were to estimate the possibility for toxicity test and compare acute toxicity of potassium dichromate in the larva stage of Neocaridina denticulata, Daphnia magna and the juvenile stage of Oryzias latipes. N. denticulate, a freshwater shrimp lives in Korea, is an indigenous species and considered to be useful for toxicity test. D. magna and O. latipes were recommended as a test species for the OECD test guideline. The 96 h-$LC_{50}$ potassium dichromate value was 0.62 mg $L^{-1}$ for the larva stage of N. denticulata and 168.44 mg $L^{-1}$ for the juvenile stage of O. latipes. The 48 h-$EC_{50}$ value was 1.27 mg $L^{-1}$ for the D. magna. The study was confirmed higher sensitivity of the larva stage of N. denticulata to potassium dichromate compared to the D. magna and the juvenile stage of O. latipes.

• Fisheries and Aquatic Sciences
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• v.19 no.9
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• pp.41.1-41.7
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• 2016

Background: The transgenic approach using estrogen-responsive regulator in fish has been given much attention as a potential means to detect and/or address estrogen-related aquatic pollutions. In order to address the development stage- and reproduction status-dependent expression patterns of the chgH-rfp transgene (red fluorescent protein transgene driven by choriogenin H promoter) in marine medaka Oryzias dancena, naturally occurring red fluorescent protein (RFP) signals under non-exposed conditions as well as the transgenically induced RFP signals under estrogen-exposed conditions were assayed. Results: Female transgenics begun to show naturally occurring RFP signals from the age of 7 weeks post hatching (WPH) without experimental estrogen exposure. Afterward, these RFP signals in female transgenics became robust with the progress of ovarian maturation. On the other hand, male transgenics did not show any naturally occurring RFP signal under non-exposed conditions irrespective of developmental stages and maturation statue. Upon exposures using estradiol-$17{\beta}$ (E2) and $17{\alpha}$-ethinylestradiol (EE2), RFP signals were significantly induced specifically in the livers of transgenic males. Conclusions: Male chgH-rfp transgenics were able to keep the "off" state of RFP expression during their entire life cycle unless exposed to exogenous estrogens. Owing to their tight regulation capability of estrogen-responsive transgene, transgenesis of chgH-rfp in male marine medaka could offer a useful model system for future ecotoxicogenomic studies regarding estrogenicity-related issues in aquatic and marine environments.

• Development and Reproduction
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• v.20 no.4
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• pp.305-314
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• 2016

The marine medaka, Oryzias dancena is a suitable sample as a laboratory animal because it has a small size and clearly distinguishes between female and male. Data on the growth and maturity of the diploid and triploid sea cucurbit species suitable for laboratory animals are very useful for studying other species. Triploidy was induced in the marine medaka by cold shock treatment ($0^{\circ}C$) of fertilized eggs for 45 min, applied two minutes after fertilization. The diploid and triploid male fish were larger than their female counterparts (P<0.05), and the concentrations of thyroid stimulating hormone (TSH) and thyroxine (T4) were higher in the induced triploids over 1 year (P<0.05). In both the diploid and tri-ploid groups the concentrations of TSH and T4 were higher in the male fish than in the females (P<0.05), while the testo-sterone and estradiol-$17{\beta}$ concentrations in the induced triploids were lower than in the diploids (P<0.05). The gonadosomatic index (GSI) of the triploid fish was lower than that for the diploids, and the GSI for females in each ploidy group were higher than that for the males. For both groups the GSI was highest at 4 months of age, and decreased thereafter to 12 months. Analysis of the gonads of one-year-old triploid fish suggested that the induction of triploidy probably causes sterility in this species; this effect was more apparent in females than in males.

• Fisheries and Aquatic Sciences
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• v.13 no.3
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• pp.224-230
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• 2010

Growth hormone (GH) is known as one of the main osmoregulators in euryhaline teleosts during seawater (SW) adaptation. Many of the physiological actions of GH are mediated through insulin-like growth factor-I (IGF-I), and the GH/IGF-I axis is associated with osmoregulation of fish during SW acclimation. However, little information is available on the response of fish IGF-II to hyperosmotic stress. Here we present the first cloned IGF-I and IGF-II cDNAs of marine medaka, Oryzias dancena, and an analysis of the molecular characteristics of the genes. The marine medaka IGF-I cDNA is 1,340 bp long with a 257-bp 5' untranslated region (UTR), a 528 bp 3' UTR, and a 555-bp open reading frame (ORF) encoding a propeptide of 184 amino acid (aa) residues. The full-length marine medaka IGF-II cDNA consists of a 639 bp ORF encoding 212 aa, a 109 bp 5' UTR, and a 416 bp 3' UTR. Homology comparison of the deduced aa sequences with other IGF-Is and IGF-IIs showed that these genes in marine medaka shared high structural homology with orthologs from other teleost as well as mammalian species, suggesting high conservation of IGFs throughout vertebrates. The IGF-I mRNA level increased following transfer of marine medaka from freshwater (FW) to SW, and the expression level was higher than that of the control group, which was maintained in FW. This significantly elevated IGF-I level was maintained throughout the experiment (14 days), suggesting that in marine medaka, IGF-I is deeply involved in the adaptation to abrupt salinity change. In contrast to IGF-I, the increased level of marine medaka IGF-II mRNA was only maintained for a short period, and quickly returned a level similar to that of the control group, suggesting that marine medaka IGF-II might be a gene that responds to acute stress or one that produces a supplemental protein to assist with the osmoregulatory function of IGF-I during an early phase of salinity change.

• Fisheries and Aquatic Sciences
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• v.20 no.9
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• pp.21.1-21.10
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• 2017

The aim of this study was to assess visible implant fluorescent elastomer (VIE) tagging and stress response in marine medaka, Oryzias dancena. The experimental fish were anesthetized individually and marked with red, yellow, or green elastomer at each of the following three body locations: (1) the abdomen, (2) the back, and (3) the caudal vasculature. During 12 months, the accumulated survival rates of fish in the experimental treatments were not different among red, yellow, and green elastomers. The experimental fish retained > 85% of the tags injected in the back, > 70% of the tags injected in the caudal vasculature, and > 60% of the tags injected in the abdomen (P < 0.05). An important observation was that the abdomen site was associated with poor tag retention. For all injected sites, the red and green tags were able to be detected more easily than the yellow tags when observed under both visible and UV lights. Tag readability was lower for the abdomen site than for the other sites (back and caudal vasculature). Thus, VIE tags were easy to apply to marine medaka (< 1 min per fish) and were readily visible when viewed under UV light.

• Fisheries and Aquatic Sciences
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• v.15 no.3
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• pp.221-231
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• 2012

The marine medaka Oryzias dancena choriogenin H gene (odChgH) and its mRNA expression during estradiol-$17{\beta}$ (E2) exposure were characterized. At the amino acid level, the choriogenin H protein is predicted to possess the conserved repetitive N-terminal region, as well as zona pellucida (ZP) and Trefoil factor family (TFF) domains. At the genomic level, odChgH has an eight-exon organization with a distribution pattern of transcription factor binding sites in the 5'-upstream region, which is commonly found in other estrogen-responsive genes. The tissue distribution pattern of odChgH mRNA was found to be gender-specific, whereby females showed a higher expression level and wider tissue distribution than did males. During embryonic development, odChgH mRNA was robustly detected from the stage of visceral blood vessel formation. Experimental E2 exposure of males resulted in odChgH mRNA being induced not only in the liver, but also in other several tissues. The E2-mediated induction was fairly dose-dependent. The basal expression levels of hepatic odChgH mRNA were lower in males that were acclimated to 30 ppt salinity than in those acclimated to 0 or 15 ppt salinity. In contrast, the inducibility of odChgH mRNA during E2 exposure was greater in seawater-acclimated fish than in brackish water- or freshwater-acclimated fish.

• Development and Reproduction
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• v.23 no.3
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• pp.199-211
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• 2019

The sterilization effects of methylene blue (MB), formalin, and iodine on the egg of marine medaka, Oryzias dancena, were investigated for disinfecting naididae worm, Chaetogaster diastrophus through sterilization. To determine harmfulness of MB, formalin, and iodine, lethal concentrations 50 ($LC_{50}$) of three chemicals were analyzed in the eggs of marine medaka. The sterilized periods of each chemical were set at 1 hr. Sterilized rates of naididae worm in each chemical were significantly affected and increased drastically as the concentration of each chemical increased (p<0.05). Sterilization abilities of naididae worm were most effective for formalin, but survival rates of egg and hatched rates for formalin were lowest among each chemical. The $LC_{50}$ of MB over 96 hrs were 185.26, 103.84, and 127.15 ppm for adults, juveniles, and eggs respectively. The toxic effects of MB were clearly dose dependent for each life stage (p<0.05). The toxicity sensitivity of juveniles to MB was dramatically higher than that of other groups. In 48 hrs after sterilization, cortisol and glucose concentrations of the adult group with MB treatment were significantly higher than those of the adult group with no treatment (p<0.05). This research provides useful data on sterilization effect of MB, formalin, and iodine, acute toxicity in marine medaka egg and toxicity, sensitivity of life stage of MB in marine medaka.

• Journal of fish pathology
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• v.24 no.2
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• pp.131-139
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• 2011

Bisphenol A (BPA) is mainly used in the production of epoxy resins and polycarbonate plastics, which is a known endocrine disruptor and acutely toxic to aquatic organisms. In this study, estrogenic effect of BPA was investigated on hybrid between Oryzias dancena and O. javanicus (ODJ). ODJ were exposed to BPA of various concentrations (eg. 2.5 mg/L, 5.0 mg/L and 10.0 mg/L) for 56 days. The growth rate, abnormality and the ratio of female and male were observed in test group and control group. As a result, the growth was $14.7{\pm}2.0$ mm in total length (TL) in 2.5 mg/L, $13.7{\pm}2.5$ mm in 5.0 mg/L, $12.8{\pm}2.5$ mm in 10.0 mg/L in test group while it was $18.0{\pm}1.2$ mm in TL in control group which was not treated with bisphenol A. The result showed that the growth decreased as the concentration of BPA increased. The abnormality rate was 13.6% in control group, 65.4% in 2.5 mg/L, 81.3% in 5.0 mg/L and 98.1 % in 10.0 mg/L which showed increase in abnormality as an increase of BPA concentration. As a result of analyzing ratio of sex in the test group and control group, 6.0% was examined to be interspecific in controls, 76.9% in 2.5 mg/L and 100.0% in 5.0 mg/L and 10.0 mg/L. In conclusion, these results suggest that BPA has estrogenic effect on ODJ.