In order to determine the testicular cycle of the Korean brown frog, Rana coreana, the gonadosomatic index (GSI) and the changes of germ cells in testis for adult males were investigated throughout the year. The study indicated that the spermatogenesis in the seminiferous tubule of testis began in August and became most active in the month of September, and the GSI was recorded the highest and the cross area of seminiferous tubule was the widest on this period. Furthermore the seminiferous tubules at the post spawning stage appeared in testis during February, and the spermatogenesis was quiescence period of time from March to July and the GSI and the cross area of seminiferous tubule were found to be the lowest. Based on these observations, we suggest that, GSI of male Korean brown frog changes significantly between July to August, indicating the testicular cycle with discontinuous spermatogenic process, and the breeding season was confirmed to be February.
The biochemical characteristics of immunorecative atrial natriuretic peptide (irAUPP and the changes in the levels of irANP in the heart were investigated during the metamorphosis of frog tadpoles. Immunohistochemical localization of pro-ANP in cardiocfes and the presence of irAW in the peritoneal fluid of metamorphosing tadpoles were also examined. The major form of irANP in the cardiocvtes of tadpoles (Rana omurensisp was high molecular weight on gel filtration chromatographv and reverse-phase HPLC. The levels of irANP in the atrium of tadpoles were five to seven times higher than those in the ventricle. In metamorphosing tadpoles the levels of irANP in the atrium increased at stage XX, the climax of metamorphosis, and decreased at stage XXV (P < 0.051, the completion of metamorphosis. When the levels of irANP was expressed as a function of body weight of tadpoles, a continuous increase in the levels of irANP was observed from pre- to postmetamorphosis (P < 0.051. The levels of irAUP in the ventricle were found to be higher in the adult frogs than in tadpoles (R. omurensisl (P < 0.01). Pro-ANP (31-67) immunoreactivity was detected in the ventricle as well as in the atrium of tadpoles fR. nigromaculotal. The peritoneal fluid was also found to contain low molecular weight of irAUP and the levmils of irANP were 55.4 $\pm$ 9.1 pg/ml. Changes of the level of irANP at different stases of the life cycle suggest that ANP may play a role in the regulation of body fluid homeostasis of frog tadpoles during the metamorphosis.
Journal of the Korean Society of Environmental Restoration Technology
/
v.19
no.3
/
pp.103-119
/
2016
This study aims to assess habitat feature on the large-scale spawning ground of the Boreal Digging Frog Kaloula borealis in Daemyung retarding basin of Daegu, and to analyze the relationships between species abundance and meteorological factors for each habitat. Fifty-seven(57) pitfalls were installed to collect species abundance of 4 survey regions, and high-resolution satellite image, soil sampling equipment, digital topographic map, and GPS were used to develop habitat features such as terrain, soil, vegetation, human disturbance. The analysis shows that the frog is most abundant in sloped region with densely herbaceous cover in southern part of the retarding basin. In the breeding season, lowland regions, where Phragmites communis and P. japonica dominant wetlands and temporary ponds distributed, are heavily concentrated by the species for spawning and foraging. Located in between legally protected Dalsung wetands and lowland regions of the retarding basin, riverine natural levee is ecologically important area as core habitat for Kaloula borealis, and high number of individuals were detected both breeding and non-breeding seasons. Temperate- and pressure-related meteorological elements are selected as statistically significant variables in species abundance of non-breeding season in lowland and highland regions. However, in sloped regions, only a few variables are statistically significant during non-breeding season. Moreover, breeding activities in sloped regions are statistically significant with minimum temperature, grass minimum temperature, dew point temperature, and vapor pressure. Significant meteorological factors with habitat features are effectively applied to establish species conservation strategy of the retarding basin and to construct for avoiding massive road-kills on neighboring roads of the study sites, particularly post-breeding movements from spawning to burrowing areas.
Objective: To evaluate the effectiveness of the Frog appliance in three dimensions by using cone-beam computed tomography (CBCT) images. Methods: Forty patients (21 boys and 19 girls), averaged 11.7 years old, with an Angle Class II division 1 malocclusion were included in our study. They had either late mixed dentition or early permanent dentition, and the maxillary second molars had not yet erupted. All patients underwent CBCT before and after the treatment for measuring changes in the maxillary first molars, second premolars, central incisors, and profile. Paired-samples t-test was used to compare the mean difference in each variable before treatment and after the first phase of treatment. Results: The maxillary first molars were effectively distalized by 4.25 mm (p < 0.001) and 3.53 mm (p < 0.05) in the dental crown and root apex, respectively. The tipping increased by $2.25^{\circ}$, but the difference was not significant. Moreover the teeth moved buccally by 0.84 mm (p < 0.05) and 2.87 mm (p < 0.01) in the mesiobuccal and distobuccal cusps, respectively, whereas no significant changes occurred in the root apex. Regarding the anchorage parts, the angle of the maxillary central incisor's long axis to the sella-nasion plane increased by $2.76^{\circ}$ (p < 0.05) and the distance from the upper lip to the esthetic plane decreased by 0.52 mm (p = 0.01). Conclusions: The Frog appliance effectively distalized the maxillary molars with an acceptable degree of tipping, distobuccal rotation, and buccal crown torque, with only slight anchorage loss. Furthermore, CBCT image demonstrated that it is a simple and reliable method for three-dimensional analysis.
This study was executed to know the effect of three differently controlled temperature conditions on Huanren brown frog (Rana huanrensis )'s growth in 2013. We've collected nine Huanren brown frog egg's sacs on Mt. Surak ($37^{\circ}40^{\prime}55.86^{{\prime}{\prime}}N$, $127^{\circ}05^{\prime}19.99^{{\prime}{\prime}}E$) in Seoul. We put those nine egg sacs in the controlled growth chambers under low temperature (LT, $5{\pm}2^{\circ}C$), medium temperature (MT, $10{\pm}2^{\circ}C$), and high temperature (HT, $13{\pm}2^{\circ}C$) conditions with three egg sacs, respectively. We measured the eggs' hatching rate, their hatching periods, and the size of the hatched individuals. The hatching rate was higher in MT (95.6%) and the rates of the other treatments were relatively lower but very similar such as LT (82.2%) and HT (82.6%). The three hatching periods were 10 days at HT, 14 days at MT and 23 days at LT. The body sizes of the hatched individuals were biggest at MT ($7.62{\pm}0.11mm$), smallest at LT ($6.82{\pm}0.10mm$) and medium at HT ($7.19{\pm}0.15mm$) (P-value ${\leq}0.0001$). From our results, we found that the various water temperatures could be very effective to Huanren brown frog eggs' hatch and growth including their body sizes. We suggest if we study more about the growth of Huanren brown adult frogs under similar temperature conditions for a long term period, it must be very helpful for conservation study about metamorphosis rate and size of adult frog as well as we could understand about the amphibians who are adapting to the climate change.
To ascertain the existence of various adrenoceptors involved in active transport of sodium in the frog skin and to delineate their physiological roles, the influence of various adrenergic agonists and antagonists on the potential difference (PD), short-circuit current (SCC) and total skin conductance (TSC) of the isolated frog skin of Rana nigromaculata were investigated. PD and SCC were determined with Ussing's technique. Drugs were administered to the serosal side of the skin. Experimental results were summarized as follows: 1. The responses to norepinephrine (NE, $6{\times}10^{-8}-6{\times}10^{-5})M$), phenylephrine (PE, $5{\times}10^{-6}-5{\times}10^{-4}M$) and epinephrine (Epi, $5.5{\times}10^{-7}-5.5{\times}10^{-5}M$) were characterized by marked elevation of PD & SCC in dose-related fashion, but the maximal effect attained by Epi was less than those of NE and PE. 2. These increments of PD & SCC were significantly inhibited by prazosin $(2{\times}10^{-6}M)$, a speciflc ${\alpha}_1$-adrenoceptor blocker. The stimulatory effect on PD & SCC were completely abolished by phenoxybenzamine (PBZ, $3.3{\times}10^{-5}M$), an irreversible ${\alpha}$-adrenoceptor blocking agent. Furthermore, with a larger doses of Epi produced marked decline of PD & SCC after the PBZ pretreatment. 3. Isoproterenol (ISP), a ${\beta}$-adrenoceptor agonist, in concentrations ranging from $5{\times}10^{-7}$ to $5{\times}10^{-6}M$ produced dose-related decrease in PD & SCC, which could be abolished by pretreatment with propranolol $(4{\times}10^{-6}M)$, a specific ${\beta}$-adrenoceptor blocker. It was further noted that the effects of Epi on PD & SCC were markedly potentiated by Propranolol pretreatment. 4. Clonidine as well as guanabenz produced increases in PD & SCC and these effects were inhibited more specifically by prazosin pretreatment than by yohimbine. These results indicated that there exist in the frog skin two distinctive types of adrenoceptors, ${\alpha}$ and ${\beta}$, which roughly corresponds to those in mammals, and that the ${\alpha}$ type of adrenoceptors mediate the stimulation of PD & SCC, whereas ${\beta}$-adrenoceptors mediate the inhibition. However, based on evidence at hand, no conclusion could be drawn on the subtype of ${\alpha}$-adrenoceptors which is involved in the stimulation of sodium transport in the frog skin.
There are many ongoing studies of infectious diseases as the major factor responsible for global declining of the amphibian population. Although some point out the amphibian rearing facilities like frog farms as one of the important sources of harboring and spreading amphibian infectious pathogens in the wild, there have been few related studies in South Korea. In this study, we investigated the bacterial and fungal colonies on the skin and in the internal organs of frogs and tadpoles collected inside and outside of Dybowski's brown frog farms in Inje, Goesan, and Gongju to compare the difference according to the region and between inside and outside the farm. We also intended to classify the bacteria collected from the tadpoles into species by analyzing 16s rDNA gene sequences. The result showed that the number of bacterial colonies found in the skin and gut of frogs and the number of fungal colonies found in the skin and liver of frogs collected in Goesan was significantly greater than those in the frogs in Inje. However, there was no difference between the frogs collected inside and outside of farms in both regions. In the case of tadpoles, the number of fungal colonies in the tadpoles collected from Gongju was greater than that in the tadpoles collected from Inje. The comparison of inside and outside frog farms showed that there were more bacterial colonies on the skin of the tadpoles collected from inside than outside the frog farm in Inje and more bacterial colonies in the organs of the tadpoles collected from outside than inside the farm in Gongju. The frogs with higher condition factor (body weight/snout-vent length*100) showed fewer bacterial colonies on the skin and fewer fungal colonies in the heart, but there were no significant relationships in tadpoles. We identified the total of 15 genera and four phyla of bacteria, but the difference according to regions and between inside and outside farm was not evident. The result of this study indicates that the different conditions according to the locality of farm and between inside and outside farm cause the difference in the population sizes of bacterial and fungal colonies and that it can affect the overall health condition of Dybowski's brown frogs in the farm. Moreover, the result suggests that effective disease control in the facility is greatly necessary to ensure successful operation of amphibian rearing facility and to prevent the possible spread of diseases from the facility to the wild.
In oreder to know the effect of the oocyte maturation with several heavy metal ions(Cd$^{2+}$, Hg$^{2+}$, Cu$^{2+}$, Pb$^{2+}$) on frog(Rana dybowskii) by FPH(Frog pituitary homogenate: 0.1 p.e./ml) in vitro, oocytes were cultured for 20 hours and exanimed the maturation rates with exposure of various comcentrations of those ions. The results showed that Cd$^{2+}$ at concentration of 0.1 ppm suppressed the maturation of the oocytes, while Hg$^{2+}$, Cu$^{2+}$ and Pb$^{2+}$ suppressed them significantly at 1 and 5 ppm respectively. To examine the reversibility of the inhibitory effects, the oocutes were exposed to the metal ions only for 3 hours and transferred to plain medium and cultured further for 17 hours. The oocytes were recovered from the toxic effect of the ions when they were exposed to 1 ppm of Cd$^{2+}$ for 3 hours and not available to 2.5 ppm. The effect of 2.5 ppm of Hg$^{2+}$, Cu$^{2+}$, Pb$^{2+}$ were also reversible and not available to 5 ppm for 3 hours exposure. From the abave results, it reveal that heavy metal ions in this study suppressed the maturation of oocytes at relatively low concentration. Therefore the oocyte culture system can be used as a useful tool to evaluate the toxicity of the pollutants in envirment.
The Frog Embryo Teratogenesis Assay-Xenopus(FETAX) protocol was recently adopted as a valuable tool fur evaluating chemical toxicity and the effect of environmental contaminants in frogs. In this study, the teratogenecity of NiCl$_2$, Carbofuran, Diazinon were determined in the Korean frog, Rana dybowskii using the FETAX protocol. The mortality rate and the percentage of malformed larvae were investigated by probit analysis, The teratogenic concentrations(EC$_{50}$) of NiCl$_2$, Carbofuran and Diazinon were 0.4 and 1.6 and 1.9 mg/1. The embryolethal concentrations(LC$_{50}$) of NiCl$_2$, Carbofuran and Diazinon were 17.6 and 41.5 and 20.2mg/1. The teratogenic indices (TI=LC$_{50}$/EC$_{50}$) were 43.8 for NiCl$_2$, 26.0 for Carbofuran and 10.6 for Diazinon. NiCl$_2$, Carbofuran and Diazinon were shown to be potent teratogens for Rana dybowskii embryo, causing concentration related increase of edema, tail and abdomen. The study results reveal that NiCl$_2$, Carbofuran and Diazinon suppress the development of embryos at relatively low concentrations. Therefore, the Rana dybowskii embryo teratogenesis assay system was proven to be a useful tool to evaluate the toxicity of environmental pollutants.lutants.
In order to clarify intra-and inter-specific variation of Korean Rana species, the partial DNA sequences of mitochondrial 16S rDNA gene were determined from 6 Korean and 1 Japanese Rana species, DNA sequences from Korean and Japanese species were comparison-analysed within, and also with the sequences from three species of Japanese brown frogs. DNA similarities were calculated as 91.3∼97.3% among brown frog (R. amurensis coreana, R. dybowskii and R. huanrenensis), as 96.11∼97.26% among pond frogs (R. nigromaculata and R. planeyi chosenica). Genetic distance of pond frog and wrinkle fyog (R. rugosa) were near than that of pond frog and brown frog. Two clusters were formed brown frogs and the other group by neigh-bor-joining and maximum-likelihood analysis, also the populations of R. nigromaculata were well distinguished between Korean peninsula and Korean island. But result from maximum-likelihood analysis slightly differed from neighbor-joining to cluster of R. rugosa. Further analyses for their population will be necessary to study the phylogenetic status.
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