MSPD 와 HPLC 를 이용한 어류의 잔류 설파제와 테트라사이클린계 항생물질의 동시분석

하대식,김종수,김곤섭 한국식품위생안전성학회 1997 한국식품위생안전성학회지 Vol.12 No.2

A simple, rapid and simultaneous analytical method is described for the detection of Sulfonamide and Tetracycline residues, i.e., Sulfamerazine (SMR), Sulfamethazine (SMT), Sulfamonomethoxine (SMM), Sulfadimethoxine (SDM), Sulfaquinoxaline (SQN), Oxytetracycline (OXY), Tetracycline (TC), Chlortetracycline (CTC). Blank control and sulfonamide and tetracycline fortified fish muscle samples (0.5 g) were blended with octadecylsilyl (C,e, 40 gm, 21% load, 60Å) derivatized silica packing material (2 g). Blended fish samples were washed with hexane, then, benzene and dichloromethane were used for the elution of tetracycline and sulfonamide, respectively, The eluants containg tetracycline and sulfonamide were analyzed by HPLC. Correlation coefficients of standard curves for individual sulfonamide and tetracycline isolated from fortified samples were linear (0.9993±0.0003-0.9997±0.0003, 0.9493±0.078-0.9753±0.036), respectively, The average percentage recoveries of sulfonamide and tetracycline ranged as 80.86-96.52% to 85.88-92.23%, and 30.01-37.12% to 65.89-73.40%, for the concentration range (0.1--1.0 ppm) examined, respectively. Limit of detection for sulfonamide was 0. 05 ug/g, then, tetracycline was 0.1 ug/g. Detection of quantitation of sulfonamide residue was 0.0012 ppm for SMR in Paralichthys Odiuacleus and 0.0020 ppm for SMR, 0.015 ppm for SMM in Cyprinus Carpio. The applicability of this procedure is demonstrated by separation and detection of incurred tetracycline and sulfonamide residues in fish muscle tissue.

HPLC에 의한 계육의 설파메타진 잔류량 분석

하대식,김종수,김곤섭,Hah, Dae-sik,Kim, Jong-shu,Kim, Gon-sup 대한수의학회 1994 大韓獸醫學會誌 Vol.34 No.1

This study was carried out to determine the sulfamethazine residues in liver and kidney of chickens. For this experiment total 80 samples of livers and kidneys were collected at random 4 points(east area 2, west area 2) meat markets in Kyong-nam area 2 and were analysed by HPLC system. The results were as follows : 1. The average concentration of sulfamethazine residues in liver and kidney were 0.056 ppm and 0.035 ppm, respectively, the sulfamethazine residues in chicken tissue was higher in liver than kidney. 2. The sulfamethazine residues of livers were exceed 0.1 ppm in three samples and no samples were exceed than 0.1 ppm in kidney. 3. No sulfamethazine residues in liver and kidney were 14 and 25 samples respectively.

Aflatoxin에 노출된 닭에서 활성탄과 어성초의 독성완화 효과

하대식,지대해,조상래,박애라,정은희,박동엽,이국천,허정호,김종수 한국동물위생학회 2010 韓國家畜衛生學會誌 Vol.33 No.2

This study was conducted to evaluate the alleviative effects of activated charcoal (AC) and Houttuynia cordata (HC) singly or in combination in broiler chickens during aflatoxicosis. Activated charcoal (1% or 0.5%) and H. cordata (1% or 0.5%) were mixed into the diets for the ability to reduced the deleterious effects of 2.4mg total aflatoxin (AFB₁) kg-¹ diet on growing broiler chickens from 1 to 21 days of age. A total of 160 1-day-old (Hyline Variety Brown) broiler chicks were housed in eight treatment groups [Control, AFB₁, AC 1%, HC 1%, AFB₁ plus AC 1% plus HC 1%, AFB₁ plus AC 1% plus HC 0.5%, AFB₁ plus AC 0.5% plus HC 1%, AFB₁ plus AC 0.5% plus HC 0.5%] each consisting of 20 chicks. Compared to control, 2.4mg AFB1 alone treatment group significantly decreased body weight gains of chickens. The addition of mixed AC 1% and HC 1% including 6, 7 groups to the 2.4mg AFB₁-containing diet moderately reduced the adverse effects of AFB₁ on performances of chickens. The chickens consuming 2.4mg AFB₁plus AC 0.5% and HC 0.5%-containing diet showed very slightly reduced the adverse effects on investigated parameters compared to the AFB₁ only treated group. Also, the single addition of AC or HC to the AFB₁-free diet had no adverse effects in chickens. These results suggest that AC and HC mixed can reduced the aflatoxicosis in broilers and may be contribute to a solution of the aflatoxicosis problem in poultry production.

비 지질 산화손상에 대한 어성초 뿌리의 항산화 효과

하대식,김충희,김의경,강정부,김종수,Hah, Dae-Sik,Kim, Chung-Hui,Kim, Euikyung,Kang, Chung-boo,Kim, Jong-shu 대한수의학회 2007 大韓獸醫學會誌 Vol.47 No.1

Houttuynia cordata root on non-lipid oxidative damage. The antioxidative efects of methanolic (MeOH) extract of Houttuynia cordata rooton non-lipid, including liposome oxidation, oxidation of deoxyribose, protein oxidation, chelating, scavenging,and 2'-deoxyguanosine (2'dG) oxidation were investigated. Houttuynia cordata root exhibited highantioxidative effect in a liposome model system. The inhibitory effect of MeOH extract on deoxyribosedamage exhibited antioxidative effect and it afforded considerable protection against damage to deoxyribose.In addition, MeOH extract at over 300extracts exhibited metal binding ability for hydrogen peroxide. Furthermore, the oxidation of 2'dG to 8-hydroxy-2-deoxyguanosine was inhibited by MeOH extracts, and scavenging activity for hydroxyl radicalexhibited a remarkable effect. The present results on biological model systems showed that MeOH extractswas effective in the protection of non-lipids against various oxidative model systems.

경남지역 야생들쥐에서 Orientia tsutsugamushi에 대한 항체 조사 및 PCR에 의한 검색

하대식,김영훈,박정웅,박재갑,김충희,류재두,정명호,허정호,서종립,조명희,이국천,김곤섭,김의경,김종수,Hah, Dae-Sik,Kim, Young-Hoon,Park, Jung-Ung,Park, Jae-Kap,Kim, Chung-Hui,Ryu, Jae-Doo,Jong, Myung-Ho,Heo, Jung-Ho,Shu, Jong-Lip,Cho, Myung-Heui 대한수의학회 2004 大韓獸醫學會誌 Vol.44 No.2

As a part of epidemiologic investigation of tsutsugamushi disease, the wild rodents which were captured in Gyeongnam area were diagnosed with indirect immunofluorescent antibody assay (IFA) and Polymerase Chain Reaction (PCR) to find if they have an antibody against Orientia tsutsugamushi. The conclusion was drawn as followings. (1) The captured 58 wild rodents showed that the subspecies distribution of Apodemus agrarius was 86.2%, Microtus fortis was 8.6% and Crocidura lasiura was 5.2%. (2) The antibody positive rate of O. tsutsugamushi Gilliam, Karp, Karto and Boryong by IFA method was 32.0% in Apodemus agrarius among 50 wild rodents and 40.0% in Microtus fortis among 5 wild rodents, respectively. It was negative in the case of all the 3 Crocidura lasiura. (3) The antibody titers on Apodemus agrarius, Microtus fortis and Crocidura lasiura against Gilliam, Karp, Karto and Boryong were measured between 1:20 and 1:640. The antibody titer against each antigen was in the order Boryong>Gilliam>Karp. (4) O. tsutsugamushi was detected from the blood, spleen and kidney from the artificially infected mice by IFA and PCR. IFA showed the positive response between 3 and 18 days after inoculation. On the other hand, positive response was found from all the samples by PCR. (5) From PCR of the genomic DNA extracted from the blood, spleen and kidney samples of the captured wild rodents, Boryong-specific amplification product with size of 210 bp, which is particular in Boryong, was detected from spleen and kidney samples, but not detected in the blood. (6) Boryong-specific amplification product was detected from spleen and kidney samples which were obtained at 3, 6, 12, 18 and 24 days after the infection with Boryong. But, it wasn't detected from the uninfected samples. (7) From PCR of spleen and kidney samples of the captured wild rodents, it was found that positive rate of O. tsutsugamushi in Apodemus agrarius and Microtus fortis were 25.0% (4/16) and 20.0% (1/5), respectively. From the above results, it can be concluded that Apodemus agrarius resided in Gyeongnam area carried O. tsutsugamushi and PCR method might be a simple, precise, rapid and useful diagnostic tool than IFA for the diagnosis of O. tsutsugamushi.

지질 과산화에 대한 전통약용 식물의 항산화 효과

하대식,김충희,김곤섭,김의경,김종수,Hah, Dae-sik,Kim, Chung-hui,Kim, Gon-sup,Kim, Eui-gyung,Kim, Jong-shu 대한수의학회 2005 大韓獸醫學會誌 Vol.45 No.3

To assess the antioxidative activity of 12 medicinal plants on lipid peroxidation, twelves traditional medicinal plants extracted with 95% methanol were investigated the antioxidative activity using DPPH, thiocyanate acid method, and thiobarbituric acid (TBA) methods. Out of 12 medicinal plants extracted with methanol, the extraction yields of Sedum kamtschaticum was the highest values (49.46%) among them and Geranicum sibiricum, Saururus chinensis root (R), Agrimonia pilosa leaf (L), Agrimonia pilosa root was the lowest value (9.97%). Radical scavenging effect of the selected traditional medicinal plants extracted from different extract solution were examined by 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical method. Antioxidative activity of methanolic extracts was higher than those of ethanol and n-hexane extracts. Scavenging effects in Sedum kamtaschaticum (R) determined by DPPH radical showed the highest among the 12 plants. The antioxidative effects of the first four medicinal plants were similar to those of butylated hydroxyanisole (BHA), and butylated hydroxytoluene (BHT), but higher than that of tocopherol, which was used as a handled control. Antioxidative effects of each indicated concentration of the methanolic extracts on linoleic acid by thiocyanate method was the highest in Sedum kamtschaticum and followed by Geum japonicum and Agrimonia pilosa and their antioxidative effect were similar to those of BHA, and BHT, but higher than that of tocopherol. Antioxidative effects of the selected medicinal methanolic extract on linoleic acid by thiocyanate acid method were examined for 15 days. Peroxidation of control and tocopherol group occurred on days 5 and 9, respectively, but BHA, BHT, selected medicinal methanolic extract group did not occur until on day 15. Antioxidative effects of the selected medicinal methanolic extract on linoleic acid by TBA method were examined for 15 days. Antioxidative activity was similar to those obtained by thiocyanate acid method.

T-2 toxin을 투여한 닭에서 Comet assay 방법을 이용한 DNA 손상 평가와 독성

하대식(Dae Sik Hah),허정호(Jung Ho Heo),이국천(Kuk Cheon Lee),조명희(Myung Heui Cho),김국헌(Kuk Hun Kim),김충희(Chung Hui Kim),류재두(Jae Du Lue),이승환(Seung Hwan Lee),김곤섭(Gon Sup Kim),김의경(Eui Gyung Kim),김종수(Jong Shu Kim) 한국독성학회 2006 Toxicological Research Vol.22 No.2

This study was designed to evaluate the possible DNA damaging effects of T-2 toxin using an alkaline single cell gel electrophoresis (SCGE) comet assay and also to investigate toxic effects in chickens. A total of 20 chickens were used in these experiments. Graded concentrations of dietary T-2 toxin (0, 4, 8, and 16 ㎍/g of diet) were given to groups of 5 broiler chickens. In comet assay, The DNA damage was analysed by the tail extent moment (TEM) and tail length (TL), which were used as markers of DNA strand breaks in SCGE. A significant dose-dependent increase in the extent of DNA migration as well as in the percentage of cells with tails was observed after treatment with T-2 toxin (P<0.05). Treatment with the low T-2 toxin (4 μ/g of diet) induced a relatively low level of DNA damage in comparison with the high T-2 toxin (16 μ/g of diet) group. The growth rate was significantly reduced by concentrations of 8, and 16 μ/g of diet (P < 0.05). The feed conversion ratio were significantly affected by any concentrations (P < 0.05). The relative weight of the spleen, and lung was decreased by the growth inhibitory concentrations. The bursa of Fabricius, thymus, and kidney were decreased in relative weight by concentrations of 16 μ/g of diet. The relative weight of the liver and heart were unaffected. The hemoglobin (Hb), hematocrit (HCT), and mean corpuscular hemoglobin (MCH) were decreased at concentration of 16 μ/g of diet. As compared with control chickens, there was no marked change in serum components except uric acid in T-2 treated chickens. All lymphoid tissues retained atrophic and lymphoid cell depletion throughout the three weeks trial.

Aflatoxin에 노출된 닭에서 활성탄과 어성초의 독성완화 효과

하대식 ( Dae Sik Hah ),지대해 ( Dae Hae Ji ),조상래 ( Sang Rae Jo ),박애라 ( Ae Ra Park ),정은희 ( Eun Hee Jung ),박동엽 ( Dong Yeop Park ),이국천 ( Kuk Cheon Lee ),허정호 ( Jung Ho Heo ),김종수 ( Jong Shu Kim ) 한국가축위생학회 2010 韓國家畜衛生學會誌 Vol.33 No.2

This study was conducted to evaluate the alleviative effects of activated charcoal (AC) and Houttuynia cordata (HC) singly or in combination in broiler chickens during aflatoxicosis. Activated charcoal (1% or 0.5%) and H. cordata (1% or 0.5%) were mixed into the diets for the ability to reduced the deleterious effects of 2.4mg total aflatoxin (AFB1) kg-1 diet on growing broiler chickens from 1 to 21 days of age. A total of 160 1-day-old (Hyline Variety Brown) broiler chicks were housed in eight treatment groups [Control, AFB1, AC 1%, HC 1%, AFB1 plus AC 1% plus HC 1%, AFB1 plus AC 1% plus HC 0.5%, AFB1 plus AC 0.5% plus HC 1%, AFB1 plus AC 0.5% plus HC 0.5%] each consisting of 20 chicks. Compared to control, 2.4mg AFB1 alone treatment group significantly decreased body weight gains of chickens. The addition of mixed AC 1% and HC 1% including 6, 7 groups to the 2.4mg AFB1-containing diet moderately reduced the adverse effects of AFB1 on performances of chickens. The chickens consuming 2.4mg AFB1 plus AC 0.5% and HC 0.5%-containing diet showed very slightly reduced the adverse effects on investigated parameters compared to the AFB1 only treated group. Also, the single addition of AC or HC to the AFB1-free diet had no adverse effects in chickens. These results suggest that AC and HC mixed can reduced the aflatoxicosis in broilers and may be contribute to a solution of the aflatoxicosis problem in poultry production.

Evaluation of Antimicrobial Activity of the Methanol Extracts from 8 Traditional Medicinal Plants

강창근,하대식,김충희,김영환,김의경,김종수 한국독성학회 2011 Toxicological Research Vol.27 No.1

The methanol extract of 12 medicinal plants were evaluated for its antibacterial activity against Gram-positive (5 strains) and Gram-negative bacteria (10 strains) by assay for minimum inhibitory concentration (MIC) and minimum bacterial concentration (MBC). The antibacterial activity was determined by an agar dilution method (according to the guidelines of Clinical and Laboratory Standard Institute). All the compounds (12 extracts) of the 8 medicinal plants (leaf or root) were active against both Gram-negative and Gram-positive bacteria. Gram-negative showed a more potent action than Gram positive bacteria. The MIC concentrations were various ranged from 0.6 μg/ml to 5000 μg/ml. The lowest MIC (0.6 μg/ml) and MBC (1.22 μg/ml) values were obtained with extract on 4 and 3 of the 15 microorganisms tested, respectively.

Xylazine의 진정효과와 α-adrenergic 수용체 봉쇄약물의 길항효과

김충희,하대식,김양미,김종수,Kim, Chung-hui,Hah, Dae-sik,Kim, Yang-mi,Kim, Jong-shu 대한수의학회 1993 大韓獸醫學會誌 Vol.33 No.1

The central nervous system depressant effect of xylazine and xylazine-ketamine was studied in chicken and mice. Intraperitoneal injection of xylazine(1~30 mg/kg) and xylazine(1~30 mg/kg)-ketamine(100 mg/kg) induced a loss of the righting reflex in chicken and mice, respectively. These effects of xylazine were dose-dependent. The results obtained were as follows; 1. The effect of xylazine-induced depression was antagonized by adrenergic antagonists having ${\alpha}_2$-blocking activity(yohimbine, tolazoline, piperoxan and phentolamine). 2. Yohimbine was most effective in the reduction of the CNS depression by xylazine. 3. Phenoxybenzamine and prazosin did not reduced CNS depression by xylazine in both species. 4. Labetalol (${\alpha}_1$, ${\beta}_1$-adrenergic antagonist) and propranolol(${\beta}$-adrenergic blocking agent) were not effective in reducing xylazine induced depression. 5. Cholinergic blocking agents (atropine and mecamylamine), a dopaminergic antagonist (Haloperidol), a histamine $H_1$-antagonist(chlorpheniramine), a histamine $H_2$-antagonist(cimetidine), a serotonergic-histamine $H_1$ antagonist(cyproheptadine) were not effective in reducing xylazine-induced depression. 6. Xylazine-induced depression is mediated by ${\alpha}_2$-adrenergic receptors and appears not to be involved in cholinergic, dopaminergic, serotonergic or histaminergic pathways.