• Korean Journal of Environmental Biology
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• v.22 no.2
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• pp.300-307
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• 2004

The chloroacetanilide herbicide alachlor (2-chloro-2', 6'-diethyl-N-(methoxymethyl)-acetanilide) and organochlorine insecticide endosulfan (6, 7, 8, 9, 10, 10-hexachloro-l, 5, 5a, 6, 9, 9a-hexahydro-6, 9-methano-2, 3, 4-benzodioxathiepin-3-oxide)are the highly toxic agricultural chemicals. Bombina orientalis is one of the most common amphibians in the world and comprises a large proportion of their total number. B. orientalis spawns in the farming regions at Spring when the massive application of agricultural chemicals occurs. These chemicals in farmland may threaten the reproduction of this frog. Therefore, we examined the embryotoxic and survival effects of alachlor and endosulfan at various concentrations in B. orientalis embryos. The survival rates of embryos at 312h post fertilization treated with alachlor and endosulfan were decreased with concentration dependent manner. Also, developmental malformations appeared by alachlor and endosulfan in B. orientalis embryos. The malformations showed in order of frequency with bent trunk, tail dysplasia, bent tail, thick-set body and ventral blister in alachlor treated embryos. The exposure of endosulfan produced 7 types of severe external malformations with tail dysplasia, pectoral blister, bent trunk, bent tail, cephalic dysplasia, ventral blister, and thick-set body. Following exposure to alachlor and endosulfan the types of malformations were diverse, suggesting these chemicals target multiple events in embryonic and larval development in this species. These results suggested that alachlor and endosulfan were detrimental for survival and development of B. orientalis embryos.

• Korean Journal of Environmental Biology
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• v.29 no.1
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• pp.74-80
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• 2011

In this study we present a mammalian cell culture model that allows to study the effect of endocrine disruptors (EDCs) on aromatase activity of aquatic amphibian, Bombina orientalis. Bombina orientalis aromatase gene was subcloned into a mammalian expression vector and subsequently transfected to mammalian cells. Although the protein expression level of Bombina orientalis aromatase was low, it had a significant aromatase activity. When EDCs were added to aromatase transfected cells, aromatase activity was significantly decreased. We report here that this system may be used to monitor the effect of EDCs on aromatase activity of aquatic organisms.

• Journal of the Korean Chemical Society
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• v.18 no.5
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• pp.368-372
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• 1974

Dark red extract of abdominal skin of Bombina Orientalis was separated and purified with TLC, PLC(preparative thin layer chromatography) and column chromatography. Through the physical and chemical properties, visible and infrared spectral characteristics the third major pigment was identified as ${\alpha}$-cryptoxanthin(3-hydoroxy-${\alpha}$-carotene)

• Proceedings of the Korea Society of Environmental Biology Conference
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• 2004.05a
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• pp.19-19
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• 2004

• Journal of the Korean Chemical Society
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• v.18 no.4
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• pp.289-296
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• 1974

More than ten cartenoid pigments were separated from the dark red extracts of abdominal skin of Bombina Orientalis by TLC, and column chromatography, and the visible and infrared spectrum of purified carotenoids were determined. Two major pigments were identified as ${\beta}$-carotene and 4-hydroxy-echinenone respectively through physical and chemical properties, chromatographic, visible and infrared spectral characteristics.

• Korean Journal of Environmental Biology
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• v.26 no.1
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• pp.22-29
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• 2008

Bombina orientalis, or Korean fire-bellied toad is belonging to Class Amphinia, Order Anura, Family Bombinatoridae, Genus Bombina. Ecological characteristics, such as small size, slow moving and swimming, gentle attitude to other species, under laboratory and/or wild condition lead the toad as suitable experimental animals with easy handling. Seasonal reproductive biology except breeding season (usually from Apr. to late Jun.) are unknown still caused by sampling methods for hibernating specimens have not been established yet. Most species of amphibian, include B. orientalis, are declined around the world related with global warming, therefore, wild caught specimen for any purposes must be limited controled soon. Even though, some pre-studies of biotope for increasing biodiversity have been reported, construction of an artificial biotope to get natural breeding cycling specimen for study of amphibian reproduction is unique modelling and only trial. In an effort to set year-round sampling for study of reproduction of B. orientalis, we suggest protocols for planning, construction manual and SOP(Standard Operation Process) to manage amphibian biotope. During pilot operation of a designed and constructed biotope, the seasonal acclimation during winter hibernation of B. orientalis were successful. It will be helpful for studying the reproductive biology of wild amphibians in temperate region and allow to get reasonable research conditions in amphibian laboratory.

• Applied Microscopy
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• v.28 no.3
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• pp.329-343
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• 1998

Early wound healing responses in the skin of the Korean fire bellied toad, Bombina orientalis, were examined using histochemical , scanning and transmission electron microscopical techniques. Cutaneous wounds were induced by sharp razor blade , and maintained in special cages for up to 3 days. By this treatment whole parts of epithelium and dermis were profoundly affected. After injury, retraction of damaged surface of the skin was observed. At 3 hours after injury, poly-band was form by mucous substance. The wound skin consisted of mucous gland secreted mucous substance which flows into wound cavity and the wound area was sealed by mucous substance which form thick layer at 6 hours. It appeared that natural disposition showed acid and neutral mucous substance by reaction of PAS and Alcian blue (pH 2.5) staining. After formation of the poly-band, epithelial cells were formed by migration of regenerating epithelial cells. These cells moved over the wound surface by migration at 12 hours after wounding. Poly-band was much increased in thickness and migrating cells were completely formed wound closure at 24 hours post-wounding. After 72 hours regenerated epithelial layer consists of four or five cell layers which are similar to clear, granular, and spinous layer.

• Applied Microscopy
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• v.28 no.3
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• pp.379-390
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• 1998

Dermal wound healing responses in the skin of the toad, Bombina orientalis, were examined using transmission electron microscopy. At 12 hours after wounding, debridement and collagenolysis occurred in damaged dermis. Histocyte has a large nucleus and long cytoplasm process. Phagocytic vesicle and lysosome were observed in the cytoplasm. Damaged blood cells were transformed spindle to irregular shape. Autolysis was observed in their cytoplasm. Histocytes are found in poly-band. The irregularly shaped nucleus is located peripheral region in cytoplasm. At 2 days after wounding, partial aggregation of blood cells is observed. Phagocytic, activity is observed in histocyte and collagenolytic collagen fibers are scattered. Fibroblast is observed in the dermis at 3 days after wounding. Clusters of ribosomes and some short cisternae of rough-surfaced endoplasmic reticulum are found in the cytoplasm. In histocyte at 7 days post wounding, various size granules composed of moderately dense material are found the cytoplasm. In this period histocyte is round to rod in profile, with slender processes projecting from the surface. At 7 day after injury, it was observed that formation of connective tissue fibers and amorphous ground substance in regenerating skin.

• The Korean Journal of Zoology
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• v.29 no.4
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• pp.261-271
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• 1986

The cutaneous xanthophore differentiation from larvae to adult in Bombina orientalis-the Korean fire bellied toad-is studied with light and electron microscopes. General structure of adult xanthophore which is composed of many pterinosomes and a small quantity of carotenoid vesicles is forming chromatophore complex with other dermal pigment cells. And the cytoplasmic process of xanthophore is distributed just beneath the basement membrane. The first differentiated xanthophore is originated from both rER rich cells and Golgi complex rich cells before and after the feeding larval stage. Formation of the pigment granule is proceeded gradually along the sequental metamorphic stages. After the metamorphosis, rapid multiplication of pterinosome is observed and enlargement of carotenoid vesicle is appeared after hybernation. These pigment granules are seen several structures by the differentiated level.

• Korean Journal of Environmental Biology
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• v.22 no.2
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• pp.329-335
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• 2004

To develop a biomarker for the monitoring of the contamination of estrogenic endocrine disrupters in the aquatic environment, reverse transcription -polymerase chain reaction (RT-PCR) analysis of vitellogenin (Vg) mRNA expression was optimized in Bombina orientalis, a Korean red bellied toad species. Based on partial cDNA sequences of both Vg and beta actin genes of B. orientalis, specific primers for RT-PCR of Vg and beta actin mRNAs were developed. Semiquantitative RT-PCR of the Vg mRNA in liver was optimized using a beta actin mRNA as an internal control in both sexes. In female RT-PCR using $1\;\mu{g}$ of the liver cDNA resulted in a linear increment in the PCR product of Vg from 18 to 34 cycles of amplification. In male, on the contrary, the RT- PCR product was first detected at 30 cycles of amplification and a linear increment was observed from 30 to 40 cycles of amplification, suggesting that male B. orientalis expresses minute amount of Vg mRNA which is a $2^{-12}$ equivalent of female. In conclusion, the optimized protocol for semiquantitative RT-PCR analysis of Vg mRNA level in B. orientalis male liver will be useful for the environmental monitoring the xenoestrogen contamination in the freshwater environment in Korea.