Evaluation of Toxic Effects Caused by Pesticides in Escherichia coli Using Recombinant Bioluminescent Bacteria

Kim,,Jiwon,Gu,,Man,Bock 한국환경독성학회 2004 환경독성보건학회지 Vol.19 No.3

본 연구에서는 유전자 재조합 발광 박테리아를 이용하여 농약에 대한 박테리아의 스트레스 반응과 세포 독성을 분석하였다 15종류의 농약에 대하여 유전자 손상, 생물막 손상, 산화적 손상 및 단백질 손상을 측정할 수 있는 발광 박테리아와 독성 유무로 인한 세포 독성을 측정할 수 있는 발광 박테리아, 5종을 이용하여 스트레스 반응을 분류하고 세포 독성 정도를 분석하였다 그 결과, 농약의 화학적 구조가 박테리아의 스트레스 반응에 영향을 미치며, 산화과정이 진행 됨에 따라 독성의 작용 기작이 변하는 것을 확인할 수 있었다. 이와 같은 유전자 재조합 발광 박테리아를 이용한 생물체내의 독성 메커니즘에 대한 분석은 생태계 유해물질들에 의한 독성을 분석하고 예상하기 위해 적용될 수 있을 것이다.

대구지역 금호강 및 주요 지천 퇴적물의 시·공간적 독성변화

정홍배,문성환,정진애,김재현,박정규,배철한,황인영 한국환경독성학회 2001 환경독성보건학회지 Vol.16 No.4

1. 사전에 선정된 공통 조사 정점들 가운데 KㆍH4(금호대교)이후의 퇴적물 독성이 상부의 것보다 크게 나타났다. 즉, 정점 KH3과 KH4, 그리고 KH4와 KH5 사이에 독성물질 유입원이 있는 것으로 판단되었다. 2. 하천 퇴적물의 독성 수준에 영향을 주고 있는 물질의 종류와 배출원 파악이 필요하였다. 이러한 내용을 향후 계속적 연구 및 조사를 통하여 수행되어야 하는 과제라 사료된다. 3. 퇴적물 시료의 채취 시점에 따라서 동일 정점의 독성 변화가 심하였다. 유해화학물질에 의한 오염 수준과 하천 생태계의 영향을 예측하기 위해서는 하천 퇴적물을 대상으로 경시적으로 독성의 변화를 계속 관측하는 것이 필요하다고 판단되었다. In aqueous ecosystems, the level of toxicity is highly responsive dependant to multiple variables, including rainfall, sunlight, pH, adhesion, etc. Because Korea has particularly distinct wet and dry seasons, the toxicity of pollutants in rivers or streams is dependant on the sampling season and time. In order to examine the effects of rainfall on toxicity, sediment samples were collected from five sites along the Keumho river. It was found that Microlox toxicity levels were generally higher during the dry season than the wet season. It indicated that river pollutants are carried off more quickly by the water during the wet season. As a result, it was recommended that the point sources of pollutants of the Keumho river would be placed between KH3 (Paldalgyo) and KH4 (Keumhogyo), KH4 (Keumhogyo) and KH5 (Dasa).

국내 하천 퇴적물 건강성평가를 위한 Microtox 독성시험 조건확립 연구

정홍배,박정규,문성환,류태권,김소정,배철한,황인영 한국환경독성학회 2001 환경독성보건학회지 Vol.16 No.4

1. 하천 퇴적물 공극수, 증류수 추출용액 및 유기용매 추출액을 pH6.0~6.5 범위로 조절한 후 Microtox 독성을 측정하는 것이 퇴적물내의 암모니아 독성이 배제된, 유해화학물질 오염에 의한 퇴적물 독성을 평가하는 방안임을 확인하였다. 2. 퇴적물에 대해 공극수, 증류수 추출물, 유기용매 추출물 구분하여 Microtox 독성을 구하는 것이 퇴적물 내 유해화학물질의 독성발현 특성을 예측할 수 있는 기법이라고 사료된다. 3. 조사정점 내 시료 채취 지점간 독성에 편차로 인하여, 동일 정점의 반경 50~100m 내에서 최소 4지점 이상의 퇴적물을 확보하는 것이 해당 정점의 독성 대표값을 산출할 수 있는 방안이 된다고 판단된다. 4. 한강수계인 남한강 지천인 복하천과 양화천, 금강수계인 청주 미호천, 대전 갑천, 낙동강수계인 대구 금호강, 영산강수계인 황룡강 그리고 익산 만경강을 대상으로 한 예비 조사한 결과, 집중연구대상 하천으로의 조건은 금호강이 가장 우수하였다. 5. 하천 퇴적물의 증류수 추출액과 유기용매 추출액의 Microtox 독성값 간의 상호 상관성이 높았다. 그러나, 농축 과정이 용이하므로 정점간 비교를 위한 독성 측정시 유기용매 추출액을 사용하는 것이 바람직하다고 사료되었다. Six rivers were selected as preliminary screening sites to determine the test conditions of Microtox in assessing the toxicity of the sediment. In addition, a pH range of 6.0--6.5 was established in testing pore water, aqueous extracts and organic extracts. Each extractable fraction of sediment showed different toxicities. Therefore, in order to properly examine the toxicity in the sediment, all extractable fractions of sediment samples needed to be tested with Microtox. Thus, sediment samples were additionally collected from at least 4 secondary sites within 50~100 m area of the primary sampling site to reduce any variation or deviation in toxicity assessment. From all sediment toxicity data that was collected from this study, it was concluded that the Keumho river was the most polluted with the highest sediment toxicity of all the rivers analyzed and needed further detailed research on its pollution problem.

2.4-Dichlorophenoxyacetic Acid의 毒性에 對한 Ethanol의 影響

安榮根,鮮于演,鄭鍾甲,金正勳 한국환경독성학회 1989 환경독성보건학회지 Vol.4 No.2

The effects of ethanol on the toxicity of 2, 4-Dichlorophenoxyacetic acid in ICR mice were examined. The results were summerized as follows; 1. The LD_(50) of 2.4-dichlorphenoxyacetic acid sodium injected intraperitoneally in mice was approximately 367 mg/kg. When the animals were administered with ethanol, 2ml/kg or 4ml/kg the LD_(50) of 2.4-D sodium were decreased to 338 mg/kg or 32 mg/kg, respectively. 2. In the acute experimental group, spontaneous motor activity in the ANIMEX system exhibited dose-dependent decrease in mice administered with 2.4-D sodium and ethanol. 3. However, in subacute experimental group, spontaneous motor activity in the ANIMEX system exhibited dose-dependent increase in mice treated with ethanol.

랏트에 대한 Polychlorinated Biphenyl의 毒性에 미치는 α-Tocopherol과 Perilla oil의 效果

崔經鉉,金文石,黃斗煥,文宰奎,金聖五 한국환경독성학회 1988 환경독성보건학회지 Vol.3 No.2

Effects of a-tocopherol and perilla oil on the toxicity of polychlorinated biphenyls (PCB) in male rat were studied. Rats were fed ad libitum for 6 weeks with the animal diet which contains PCB 30 ppm and 100 ppm. Perilla oil (0.5 g/kg body weight) and a-tocopherol (30 mg/kg body weight) were administered intraperitoneally twice a week for 6 weeks. Rats fed with PCB showed enlargement of liver and spleen, increase in aspartate aminotransferase, alkaline phosphatase, sereum lipid and cytochrome P 450 and decrease in body weight and glutathione. When perilla oil was administered to rats fed with PCB increase in aspartate aminotransferase, alkaline phosphatase, serum lipid and cytochrome P450 and decrease in body weight and glutathione were significantly augmented, compared to rats fed with PCB alone. This means that perilla oil potentiates the toxicity of PCB. On the other hand when a-tocopherol was administered to rats fed with PCB increase in aspartate aminotransferase, alkaline phosphatase, serum lipid and cytochrome P450 and decrease in body weight and glutathione were signigicantly reduced, compared to rats fed with PCB alone. This means that a-tocopherol reduces the toxicity of PCB. From the above results, it may be concluded that PCB is metabolized by microsomal mixed function oxidase and the metabolite causes the toxicity and microsomal glutathione plays a role of protection on the toxicity of PCB.

랏트에 있어서 Vitamin A Acetate의 毒性에 미치는 α-Tocopherol의 影響

安榮根,金聖五,吳然俊,朴榮吉 한국환경독성학회 1989 환경독성보건학회지 Vol.4 No.2

Influences of α-tocopherol on the toxicity of vitamin A acetate in male rats were studied. The obtained results are as follows; 1) The administration of vitamin A acetate 500,000 IU/Kg i.p. twice at 3 days interval decreased the liver weight/body weight and increased the spleen weight/body weight, and increased activities of SGOT and alkaline phosphatase, and also increased BUN and creatinine. 2) α-Tocopherol administered together with vitamin A acetate as given as the above 1) poteniated the increase of SGOT activity caused by vitamin A acetate and reduced the increase of alkaline phosphatase activity and creatinine which were caused by vitamin A acetate. 3) The administration of vitamin A acetate 500,000IU/Kg i.p. twice a week for 4 weeks showed remarkable decrease of body weight gain and the effect of it was larger in later stage than in early. It increased significantly liver weight/body weight and further increased the activities of SGOT, SGPT and alkaline phosphatase, and showed no influnence on BUN and creatinine. 4) α-Tocopherol administered together with vitamin A acetate as given as the above 3) reduced the decrease of body weight gain caused by vitamin A acetate, and potentiated remarkably the increased activities of SGOT and alkaline phosphatase which were caused by vitamin A acetate.

Testicular Toxicity of 2-Bromopropane in Male Sprague-Dawley Rats

Cha,,Sang-Eun,Park,,Sang-Rae,Kim,,Kgu-Hwan,Choi,,Jeung-Suk,Kim,,Sung-Kuk,Jeon,,Tae-Won 한국환경독성학회 2000 환경독성보건학회지 Vol.15 No.4

택트 스위치 제조공정 침지액의 주성분인 2-bromopropane 독성에 대한 연구로 최단기의 폭로로 농도를 달리하여 3주간 반복 투여 시험을 시행하여, 흰쥐의 혈액 및 세정관의 변화를 관찰하기 위해 투여기간 동안의 체중의 변화, 고환, 간, 신장 등의 장기무게, 혈액화학과 혈액학적 분석 및 고환의 병리조직의 변화 등을 관찰하여 2-bromopropane의 급성투여 조건에서의 중독현상을 비교렉劇?臼눼? 농도를 달리하여 투여에 따른 체중변동은 통계적으로 유의한(P<0.05) 체중의 감소를 나타내었다. 1,000㎎/㎏ 투여군에서 백혈구수, 적혈구수, 혈구용적과 혈색소 농도에서 유의한 변화(p<0.05)를 보였다. 조직병리학적 소견으로 정소에서는 세정관의 정조세포와 정모세포의 괴사를 볼 수 있었고, 기저막의 비후, 세정관의 Sertoli 세포는 광범위하게 세포질성 공포현상을 보여주고 있다. 또한 간질조직에서는 Leydig 세포의 증식을 볼 수 있었다. 2-bromopropane의 손상부위는 조혈과 생식계가 표적으로 생각되며, 고농도 투여가 저농도 투여에 비해 독성이 심하며 독성물질의 양-반응 관계를 보여주고 있다. 2-Bromopropane, important industrial chemical, specially in electronic industry at Yangsan in Korea has been reported to cause amenorrhea for female and azoospermia, oligozoospermia or reduced sperm motility for male. 2-BP was investigated through 21 days of repeated dose in male Sprague-Dawley rats. The dose levels per body weight were 0 (control), 250, 500 and 1,000 mg/kg.'2-BP dissolved in vehicle olive oil was injected into the intraperitoneum 6 times per week for 3 weeks, but 1,000 mg/kg dose group was 2 weeks because of serious illness. Male rats showed significant decreases in body weight and right and left testis showed typical weight losses depending on the 2-BP. The number of white blood cell and red blood cell, percentage of monocytes, and hemoglobin decreased significantly in high dose (P<0.05). Red cell volume distribution width increased significantly in the high dose (P<0.05). Histopathological findings of testes showed a decrease of spermatogenic cells, exfoliation of spermatid and spermatocyte, vacuolization of Sertoli cells and hyperplasia of Leydig cells. Protein band density between 113,000 dalton (β-galactosidase)and 53,900 dalton (ovalbumin) has decreased in 250 mg/kg dose group, but it has gradually increased to the higher density in 1,000mg/kg dose group than in control group.

Tyrosine Hydroxylase in Japanese Medaka(Oryzias latipes) : cDNA Cloning and Molecular Monitoring of TH Gene Expression As a Biomarker cDNA 클로닝 및 생물지표로서의 TH 유전자 발현의 분자생물학적 추적

Shin,,Sung-Woo,Kwon,,Oh-Shin,Kim,,Jung-Sang,Chon,,Tae-Soo,Lee,,Sung-Kyu,Koh,,Sung-Cheol 한국환경독성학회 2000 환경독성보건학회지 Vol.15 No.4

최근 독성 유해물질의 환경으로의 방출로 인해 인간 및 생태계에 대한 위해성 문제가 심각하게 제기되고 있다. 독성화학물질을 포함한 여러 환경 오염물질의 위해성평가는 화학물질의 유해성과 노출량 측정을 동시에 측정함으로써 가능한데 이 경우 생물지표(biomarker)가 최근 각광을 받고 있다. 본 연구에서는 동물의 행동에 관련된 신경전달물질의 생성에 결정적 역할을 하는 tyrosine hydroxylase (TH) 및 그 유전자가 생물지표로서 이용 가능성이 있는지를 검토하였다. Ovary cDNA library의 PCR 스크리닝을 통한 송사리 TH 유전자를 부분적으로 클론하였으며(327bp), DNA 염기서열 분석 결과 쥐(rat)의 TH 유전자와 동일한 염기서열을 보였다. 그리고 다이아지논 처리구 및 무처리구에 송사리의 머리부분(head)및 몸통 부분(body)에서 추출된 총 RNA에 TH mRNA가 존재함을 RT-PCR를 통하여 확인하였다. 그러나 다이아지논의 처리효과가 송사리의 행동에 미치는 영향을 보기 위해서는 TH 의 발현을 보다 정량적으로 검토할 필요가 있을 것으로 판단된다. 생물지표로서 TH의 활성 및 mRNA의 기관별 또는 조직별 검출은 독성물질에 영향을 받는 어류 신경행동 변화를 모니터링 할 수 있는 유용한 수단이 될 것이다. 나아가 환경관리에 있어서 신경화학물질과 분자생물학적 상관관계를 통한 이상반응행동의 분석은 환경 위해성평가에 상당히 기여할 것이다. The release of hazardous waste materials into the environment poses serious risks in humans and ecoThe risk assessment of environmental polluincluding hazardous chemicals requires a commeasurement of hazard and exposure of the chemicals that can be achieved by toxicity evausing a biological system such as biomarkers. In this report we have tried to develop a biomarker used to elucidate a molecular basis of, and to monitor abnormal behaviors caused by diazinon in Japanese medaka (Oryzias latipes) as a model organism. First, an attempt was made to clone tyrosine hydroxylase gene from Japanese medaka that would be a candidate for a biomarker for neuronal modulations and behaviors. For monitoring experiments at behavioral and molecular biological levels, the fish were treated under different sublethal conditions of diazinon and their behavioral responses were observed. In this study we have successfully cloned a partial TH gene from the medaka fish through PCR screening of an ovary cDNA library. DNA sequencing analysis revealed that the amplified fragment was 327 by encoding 109 amino acids. Comparing the DNA sequence of medaka TH with other species, TH gene revealed the DNA sequence was completely identical to that of rat TH. In the RT-PCR, 330 by of mRNA was consistently amplified in all the treated samples including control. There were no significant differences in the TH expression level regardless of treating concentrations (1~5,000ppb) and time (0~48 hr). The reason appeared to be that RT-PCR was not performed using through a quantitative analysis normalized against an actin gene expression. Organ or tissue-specific detection of TH activity and mRNA as biomarkers will be a useful monitoring tool for neurobehavioral changes in fish influenced by toxic chemicals. Furthermore, quantitative analysis of locomotive patterns and its correlation with the neurochemical and molecular data would be highly useful in measuring toxicity and hazard of various environmental pollutants.

박테리아(E. Coli)의 AGB(alpha-glucosidase biosynthesis)를 이용한 중금속의 독성시험과 풀빅한에 의한 독성 억제효과

김재현,김명길,엄애선 한국환경독성학회 2004 환경독성보건학회지 Vol.19 No.1

Aim of the present study was the development of a bioassay which enables the detection of toxic effects of heavy metal ions to a bacterium, Escherichia coli. Inhibition effects of the metals on growth rates of the bacterium were studied in the absence or presence of fulvic acid. This method does not clearly differentiate among metals, hut does detect overall AGB inhibition rate (toxicity) for 5 different heavy metals. The toxicity of the metals in the absence of fulvic acid in the same testing conditions was significantly increased in following order: Hg<Ph, Zn<Cd<Cu, whereas the inhibition rate(toxicity) in the presence of FA was shown to be increased in following order: Cd<Ph, Hg<Cu<Zn. The results of the present study indicate that this simple and fast biomonitoring assay with direct exposure of E coli. might be a valuable supplement to analytical methods of contaminated media.

29차 환경성돌연변이 발암원학회 ( environmental mutagen society ) 를 다녀와서

류재천 한국독성학회 1998 한국독성학회보 Vol.12 No.2