• Korean Journal of Veterinary Research
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• v.35 no.3
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• pp.635-641
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• 1995

The purpose of this study was to investigate the effects of developmental stage and equilibration time on survival of rabbit embryos following freezing by vitrification. Adult New Zealand White female rabbits were superovulated with PMSG and hCG. The 8-cell stage embryos were collected from 40 to 45 hours after hCG injection by flushing oviducts with Dulbecco's phosphated buffered saline and in vitro cultured in TCM-199 containing 10% fetal calf serum(FCS). Each embryos developed in vitro to 16-cell, compact morula and blastocyst was cryopreserved and cultured following thawing to examine their developmental potential to expanded blastocyst stage in vitro. The frozen-thawed-cultured embryos were stained with Hoechst 33342, and their nuclei were counted using a fluorescence microscope. On the toxicity test of EFS solution as cryopreservation, the survival rates of 8-cell stage embryos was decreased in reverse to increasing of exposure time over 5 minutes. The post-thaw survival rates of embryos on equilibration times was significantly(P<0.05) higher for 2 min. than for 5 or 10 minutes. From morula to blastocyst of rabbit embryos was more suitable than 8-cell stage for cryopreservation by vitrification. The higher post-thaw survival rate of embryos can be achieved by keeping the cryoprotectant at $4^{\circ}C$ than at $20^{\circ}C$. The mean number of nuclei per embryo following freezing by vitrification and in vitro culture to expanded blastocyst at compacted morula and blastcyst was not significantly differ from fresh blastocyst.

• Korean Journal of Veterinary Research
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• v.34 no.2
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• pp.361-368
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• 1994

Teratogenic and embryotoxic effects of mercury have been reported, however, there is little information about possible antidotes against mercury exposure during gestation. In order to evaluate therapeutic effects of selenium as an antidote against mercury poisoning, pregnant CD-1 mice were exposed to methylmercury chloride(20ppm) through the drinking water with treatment of sodium selenite (1.0mg, 2.0mg or 3.0mg/kg b.w., subcutaneously) or BAL(5.0mg/kg b.w., subcutaneously) under the single or combination base as the therapeutic agents from day 6 to 15 of gestation. Fetal growth parameters such as body weight and crown-rump length in the mice exposed to mercury, were reduced as was placental weight compared to those in the control. Treatment of selenium(alone, combination with BAL) reduced the harmful effects induced by mercury on the fetal growth parameters even though no specific relationship between dose and therapeutic effect. The incidence of dead fetuses/resorptions and malformed fetuses(especially cleft palate) was also increased in the mercury only treated group. Selenium treatment demonostrated reduced the incidence of abnormal fetuses under the exposure of mercury. Relative maternal organ weights(liver, kidney, spleen) were increased significantly but relative brain weight was decreased as evidenced by decreased in the mercury treated mice compared to that in the control. A subtle indication of maternal mercury toxicity evidenced by changes of relative maternal organ weights, decreased water and feed consumption were also prevented efficiently by selenium treatment. The present study suggests that methylmercuric chloride is embrytoxic and teratogenic in CD-1 mice when exposured during organogenesis and that selenium administration may have therapeutic application for the treatment of mercury poisoning although more applicable study in human should be performed with caution in the future.

• Journal of Environmental Health Sciences
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• v.42 no.4
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• pp.266-273
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• 2016

Objectives: This study evaluated the virucidal efficacy against avian influenza virus (AIV) of a disinfectant spray containing 0.25% grapefruit seed extract, 0.2% citric acid, 0.0625% malic acid and 0.0125% benzalkonium chloride. Methods: The disinfectant spray was diluted several times with hard water (HW) and organic matter (OM). Two point five mL of each diluent was added into each test tube, and 2.5 mL of AIV suspension was inserted into each test tube. After 30 minutes of virus-disinfectant contact reaction at $4^{\circ}C$, 2.5 mL of 10% inactivated fetal bovine serum was added into each test tube to neutralize the sanitizer efficacy. The neutralized solutions were serial 10-fold dilutions with phosphate buffer solution, and 0.2 mL of the diluents was injected into the allantoic cavity of five ten-day-old-chickens per dilution time. After incubation of the embryos for five days, the viability of the AIV was examined by hemagglutination titer. The valid dilution of the disinfectant spray was estimated according to the dilution time that the virus titer was inactivated more than $10^4$ 50% egg-infective dose (EID50)/mL compared with pathogen control. Results: In HW and OM conditions, the valid dilutions of the disinfectant spray against AIV were seven- and three-fold dilutions, respectively. The AIV titer of the pathogen control was more than 6.1 log10EID50/mL, and there was no embryonic toxicity. Conclusion: The present study showed that this disinfectant spray has effective virucidal activity against AIV.

• Journal of Embryo Transfer
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• v.23 no.4
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• pp.263-267
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• 2008

The purpose of this study was to determine toxic effect of sucrose and trehalose prior to cryopreservation on nuclear maturation and embryonic development in immature bovine oocytes. All cryoprotectant was prepared in tissue culture medium 199-HEPES (TCM 199-HEPES) with 10% fetal bovine serum (FBS). Immature oocytes were exposed to 1.2M ethylene glycol (EG) and 0.1M sucrose or 1.2M EG and 0.1M trehalose for 3 min and then were exposed to 3.2 M EG and 0.25 M sucrose or 3.2 M EG and 0.25 M trehalose for 1 min. Oocytes treated with cryoprotectants were exposed to 0.25 M sucrose or 0.25 M trehalose for 5 min and then 0.1 M sucrose or 0.1 M trehalose for 5 min. Depending on type of sugar added to cryopreservation solution, oocytes were allocated to sucrose group and trehalose group, respectively. Oocytes exposed to TCM 199-HEPES with 10% FBS were considered as control. Oocytes were cultured in TCM 199 supplemented with 10% FBS, 5 ng/ml epidermal growth factor, 0.01 IU/ml luteinizing hormone, and $1\;{\mu}g/ml$ estradiol for 24 h in $39^{\circ}C$, 5% $CO_2$. Nuclear maturation was assessed by staining oocytes with 1% aceto-orcein. Oocytes were fertilized in vitro and were cultured in TCM 199 supplemented with 10% FBS, 5 mM sodium pyruvate, and antibiotics in $39^{\circ}C$, 5% $CO_2$. The rates of cleavage and blastocyst, and cell number in blastocyst were assessed. Metaphase II rates were not different among experimental groups regardless of type of sugar. The cleavage rate of trehalose group (73.3%) was significantly higher (p<0.05) than those of sucrose group (62.8%) and control group (60.8%). The blastocyst rate was significantly higher in trehalose group (p<0.05). Mean cell number in blastocyst were not different among experimental groups, although cell number of blastocyst in trehalose group was significantly higher on day 7 (p<0.05). In conclusion, sucrose and trehalose were not toxic to immature bovine oocytes prior to cryopreservation. In particular, trehalose was more effective on embryonic development.

• Journal of Environmental Health Sciences
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• v.33 no.4
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• pp.306-316
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• 2007

This study have a object to found out the effects of oriental herb medicine, Antaeeum, to dams of rats and their offsprings. The Antaeeum was savaged to female Sprague-Dawley rats at a dose of 5 mg/kg/day for 3 weeks during gestation periods. Dams of rat were sacrificed at 20th day of gestation, and were observed major internal and reproductive organs. Approximately live fetuses in the 20th days of gestation were selected randomly and examined with stereo microscopes. Others offsprings were fixed with 95% ethanol for skeletal examinations. The fixed fetuses were stained with alcian blue and alizarin red S to observe skeletal variations or malformations. Maternal body weight of Antaeeum treated dams have a tendency of increasing compared with control dams. There were no significant difference in internal and reproductive organs of weight or findings. The spleenic organ relative weight of treated dams were decreased compared with the control significaltly (p<0.05). There were no significant changes between two groups in blood chemistry and hematological values. There were no significant changes in number of corpus luteum, implantation, live fetuses and implantation rate, delivery rate, late resorption rate and sex ratio. But in the Antaeeum treated group showed lower early resorption rate than that of the control dams. Fetal body weight and number of fetus a dam at Antaeeum treated group were higher than that of control group. The fetuses of dams treated with Antaeeum didn't induced external malformations. Vertebral and sternal variations were observed in Antaeeum group, but compared with the control, those variations were not significant. The ossification numbers of rib, cervical, thoracic, and lumber were normal. Fetuses treated with Antaeeum to the dams showed no significant difference in the number of caudal vertebra (P>0.01). From these results, it can be concluded that Antaeeum showed no toxicity effects on maternal side especially on body weight, early resorption rate, and number of live fetuses. Also there were no significant changes on maternal organ weights except spleen, hematological data, reproductive organs. Although skeletal variations were examined at vertebra and sternum, this Antaeeum could not induced significant choses in bone malformation.

• Journal of Environmental Health Sciences
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• v.38 no.1
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• pp.8-17
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• 2012

Objectives: Perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) are man-made, persistent global pollutants widely diffused throughout the environment. They have been even found in the cord blood and breast milk of humans. Furthermore evidence of developmental toxicity in animals exists. To assess the distribution of maternal and fetal exposure to PFOS and PFOA, we analyzed paired maternal blood, cord blood and breast milk samples. Methods: Maternal blood, cord blood and breast milk were collected from 150 volunteers from the general population (aged 20-40, mean $30.5{\pm}2.9$) of the city of Busan in 2009-2010. The samples were extracted using the weak anion exchange and solid-phase extraction methods and quantified by high-performance liquid chromatograph (HPLC, Agilent 1200 Series) coupled with an Triple Quad LC-MS/MS system (Agilent 6410). Results: Median PFOA and PFOS concentrations in maternal blood were 2.18 and 3.32 ng/ml, in cord blood were 0.83 and 0.58 ng/ml, and in breast milk were 0.13 and 0.11 ng/ml, respectively. PFOS and PFOA concentrations were significantly correlated among matrices (Spearson's ${\rho}=0.226$, p = 0.05 for maternal blood; ${\rho}=0.736$, p < 0.01 for cord blood; ${\rho}=0.493$ p < 0.01 for breast milk). The ratio of cord blood/maternal blood was 0.39 for PFOA and 0.19 for PFOS. The ratio of breast milk/maternal blood was 0.07 for PFOA and 0.06 for PFOS. Conclusions: Our findings suggest that PFOA and PFOS exposure through the placenta was more prominent than through breast milk among Korean neonates born in Busan. The transfer efficiency of maternal blood to breast milk was similar between PFOA and PFOS, but that of maternal blood to cord blood was higher in PFOA than PFOS.

• Journal of Environmental Health Sciences
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• v.31 no.6
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• pp.457-466
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• 2005

Industrial plasticizers such as phthalates can induce peroxisome proliferation. A growing concern among environmental and health groups has arisen because phthalates such as di-2-ethy1hexy1 phthalate (DEHP) and DBP may cause hormonal disorders, reproductive toxicity, hepatocellular tumors, genital disorders owing to a capacity to bind estrogen receptors, and a low-dose toxic action during certain periods of fetal development. Phthalate esters are used extensively as a plasticizer for plastic manufacture such as PVC bags and medical devices. This study investigated the effects of leached components from spring water container's cap and seal film. Phthalates, e.g. dimethy1 phthalate (DMP), diethy1 phthalate (DEP), di-n-buty1 phthalate (DBP), benzy1buty1 phthalate (BBP), di-(2-ethy1hexy1) phthalate (DEHP), and bisphenol A (BPA) were measured in the spring water. The bisphenol A was not detected or below the detection limit on the leaching from cap, sealing or spring water. DEHP were detected 90-116 ppb on the leaching from seal after 2 weeks, and 0.48-0.51 ppb from the spring water after I week. BBP were measured from seal within 1 week 25.4-66 ppb and below 0.12 ppb from spring water within 2 days. DMP were detected from seal within 2 weeks 51-68.5 ppb and 0.12 ppb after 2 weeks. DEP were measured from seal within 2 weeks 48.1-141 ppb and the concentrations were increased by the time from 0.10 to 0.31 ppb at spring water. DBP were detected from the seal within 2 weeks 92.3-5100 ppb and the concentration were decreased by the time from 0.24 to 0.10 ppb at spring water. These results indicate that some phthalate esters contaminated with spring water using the intact cap and seal film. It is concluded that the measured levels of phthalates leaching from these materials might in vivo only be slightly less than 1/10 of the LOAEL.

• Korean Journal of Veterinary Research
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• v.34 no.1
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• pp.165-172
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• 1994

DA-125 is a new anthracycline antitumor antibiotic, which is derived from adriamycin. The potential of DA-125 to induce embryotoxicity was evaluated in the Sprague-Dawley rats. One hundred twenty naturally mated SD rats(sperm in vaginal lavage=day 0) were distributed among three treated groups and a control group. DA-125 was administered intravenously at dose levels of 0. 0.1, 0.3 and 1.0mg/ kg/day. Dams were treated from day 7 to 17 of gestation and were subjected to the caesarean section on day 20. At 1 mg/kg, reduced food intake, reduced body weight and decreased weight of spleen were observed in dams. An increase in the resorption rate and a reduction in the fetal weight were also found. In addition, various types of external, visceral and skeletal malformations occurred at an incidence of 11.9, 41.8 and 14.5%, respectively. Characteristic malformations include exencephalia, gastroschisis, cleft lip, dilatation of lateral and 3rd ventricle, fused ribs, among others. There were no signs of maternal toxicity or embryotoxicity at 0.1 and 0.3mg/kg. The results show that the test agent DA-125 is embryotoxic at maternally subtoxic dose in rats.

• Journal of Food Hygiene and Safety
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• v.27 no.1
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• pp.74-80
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• 2012

Fish contain both the neurotoxin methyl mercury (MeHg) and nutrients important for brain development. The developing brain appears to be most sensitive to MeHg toxicity and mothers who consume fish during pregnancy expose their fetus prenatally. Although brain development is most dramatic during fetal life, it continues for years postnatally and additional exposure can occur when a mother breast feeds or the child consumes fish. This raises the possibility that MeHg might influence brain. We evaluated the relationship between fish consumption and mercury exposure levels in umbilical cord blood of the pregnant women of the city of Tongyeong city, Korea. A total of 159 pregnant women residing in the city of Tongyeong, Korea were recruited for the study between October 2010 and March 2011. Fish consumption was evaluated using food frequency questionnaires including detailed questions on fish consumption. We used ANOVA to estimated the particular relevance between the frequency of fish consumption and the umbilical cord blood mercury concentration, and other various factors. The average mean concentration of mercury levels in umbilical cord blood of pregnant women who participated in our study were $2.69{\pm}2.50ppb$, ranging from 0.01 to 14.80 ppb. The mean concentration of umbilical cord blood mercury exposure was lower than the level recommended by WHO (5.0 ppb), but the mercury exposure level exceeded the WHO recommended in 17 (10.7%) cases of umbilical cord blood. Mercury levels in cord blood of pregnant women were $2.04{\pm}2.00ppb$, ranging from 0 to 8.00 ppb in below 29 years old and $3.18{\pm}2.74ppb$, ranging from 0.01 to 14.80 ppb in more 30 years old. In this study, there was a significant difference for the frequency of eating fish between the groups (p < 0.01). The level of the groups that ate fish 3 to more times per week ($4.15{\pm}4.02ppb$) was significant higher as compared with the level of other groups that ate fish 1 to times per week ($2.63{\pm}2.22ppb$) and none per week ($1.06{\pm}1.44ppb$), respectively. We found that the mercury concentration of umbilical cord blood associate with fish consumption and this was statistically significant and this fact revels that fish consumption is positively related to mercury levels in the umbilical cord blood. We need systematic and periodic research on the general population to prevent mercury poisoning, which can be cause by low-level mercury exposure from dietary intake such as chronic fish consumption.