본 연구에서는 상수원지(영천호, 안계호, 가창호, 팔당호)에서 남조류가 생성하는 간장독소인 microcystin 생성특성을 조사 하였으며, 상수원수와 어류의 체내 존재하는 microcystin의 위해가능성을 알아보고 Microcystis 종들의 세포당 microcystin 농도에 근거한 위해성 기준을 제안하고자 하였다. 조사대상 상수원지에서 Microcystis 5종(...
본 연구에서는 상수원지(영천호, 안계호, 가창호, 팔당호)에서 남조류가 생성하는 간장독소인 microcystin 생성특성을 조사 하였으며, 상수원수와 어류의 체내 존재하는 microcystin의 위해가능성을 알아보고 Microcystis 종들의 세포당 microcystin 농도에 근거한 위해성 기준을 제안하고자 하였다. 조사대상 상수원지에서 Microcystis 5종(M. aeruginosa, M. ichthyoblabe, M. flos-aquae, M. wesenbergii, M. novacekii)과 Anabaena 2종(A. spiroides 및 A. sp.)이 출현하였다. 이들 남조류는 영천호에서 4월 ~ 12월, 안계호와 가창호에서 4월 ~ 11월 사이에 발생하였으며, 팔당호에서는 6월과 7월에만 관찰되었다. 분자생물학적 방법(TOX2P/TOX2F primer set)을 이용한 실험에서 팔당호를 제외한 세 개의 호수(영천호, 안계호, 가창호)에서 모두 microcystin 농도가 검출되지 않은 남조류 발생초기인 4월과 5월에 mcy 유전자가 확인되었다. HPLC법을 이용한 microcystin의 정성ㆍ정량적 분석에서 영천호는 microcystin(-RR, -YR, -LR)이 6월 ~ 12월(9.5 μgㆍg-1 ~ 821.1 μgㆍg-1), 안계호는 6월 ~ 11월(40.3 μgㆍg-1 ~ 356.3 μgㆍg-1), 가창호는 6월 ~ 11월(44.6 ~ 169.3 μgㆍg-1)에 microcystin이 각각 검출되었으며, 이들 상수원지에서 모두 공통적으로 microcystin-RR과 -LR의 검출빈도가 높았으며, 특히 microcystin-RR의 농도가 가장 높았다. 영천호, 안계호 및 가창호에서 M. aeruginosa는 microcystin 농도와 가장 높은 상관성을 나타내는 주요 독성종으로 확인되었으며, 총인과 총질소 등의 영양염이 M. aeruginosa의 증식을 유발함으로써 간접적으로 microcystin 생성의 증가에 영향을 주는 것으로 나타났다. 현장에서 분리한 M. aeruginosa 균주들의 microcystin 생성비율(75%)은 다른 Microcystis 균주들 보다 높았으며, microcystin 정성·정량적 분석결과 또한 현장의 microcystin 생성패턴과 매우 유사하였다. 높은 농도의 microcystin 생성이 관찰된 M. aeruginosa YC, M. ichthyoblabe AK와 M. viridis AK는 배양실험 조건에서 최적의 성장 온도(30 ~ 35℃)보다 낮은 20℃와 10 ~ 20 mgㆍL-1의 질소 농도에서 가장 높은 microcystin 생성을 나타내었다. M. aeruginosa YC는 120 μmolㆍm-2ㆍs-1의 조도와 0.5 mgㆍL-1의 인 농도에서 microcystin 생성의 최대값을 나타내었다. 반면에, M. ichthyoblabe AK와 M. viridis AK는 각각 70 μmolㆍm-2ㆍs-1와 35 μmolㆍm-2ㆍs-1의 조도와 5 mgㆍL-1의 인 농도에서 microcystin 생성의 최대값을 나타내었다. 우리나라의 조류예보제 기준(남조류 세포수)과 WHO (1998)의 microcystin-LR 가이드라인(1 μgㆍL-1)에 기초하여 상수원수내 microcystin의 위해성을 검토한 결과, 영천호와 안계호는 위해가능성이 있는 것으로 간주하는 조류경보 단계(5,000 cellsㆍmL-1 초과)가 각각 6개월(6월 ~ 11월)과 3개월(9월 ~ 11월)동안 지속된 반면, 가창호는 5월부터 10월까지 위해성이 없는 것으로 간주하는 조류주의보 수준(500 cellsㆍmL-1 초과, 5,000 cellsㆍmL-1 미만)을 유지하였다. 하지만, WHO (1998)가 정한 가이드라인에 기초했을 때, 가창호는 6월 ~ 11월에 최대 4배 초과하였으며, 안계호는 6월 ~ 11월에 약 1배 ~ 8배 초과하였다. 반면에, 영천호는 8월 ~ 11월 사이에 2배 ~ 33배 초과하였다. 더욱이 Falconer et al. (1994)이 제시한 세포당 microcystin 농도 계산식을 적용한 가창호(M. aeruginosa A3, M. aeruginosa JE10)와 영천호(M. aeruginosa SE20) 균주들의 실험결과들은 우리나라의 조류경보수준(5,000 cellsㆍmL-1)의 1/2 이하인 약 2,000 cellsㆍmL-1에서 독소 가이드라인인 1 μgㆍL-1의 microcystin 농도와 일치하여 M. aeruginosa를 중심으로 조류경보수준의 재검토가 필요한 것으로 조사되었다. 영천호에서 채집한 어류들의 체내 축적된 microcystin이 인체에 미치는 잠재적 위해성을 알아보았다. TDI (total daily intake) 값과 비교했을 때 어류들은 모두 간에서 EDI (estimated daily intake) 값이 TDI 값을 모두 초과하였으며, 떡붕어의 간에서 EDI 값이 가장 높게 나타났고 TDI 값의 약 8배 초과하였다. 따라서, 영천호에서 남조류 대발생시 떡붕어 섭취에 의한 위해가능성이 확인되었다.
본 연구에서는 상수원지(영천호, 안계호, 가창호, 팔당호)에서 남조류가 생성하는 간장독소인 microcystin 생성특성을 조사 하였으며, 상수원수와 어류의 체내 존재하는 microcystin의 위해가능성을 알아보고 Microcystis 종들의 세포당 microcystin 농도에 근거한 위해성 기준을 제안하고자 하였다. 조사대상 상수원지에서 Microcystis 5종(M. aeruginosa, M. ichthyoblabe, M. flos-aquae, M. wesenbergii, M. novacekii)과 Anabaena 2종(A. spiroides 및 A. sp.)이 출현하였다. 이들 남조류는 영천호에서 4월 ~ 12월, 안계호와 가창호에서 4월 ~ 11월 사이에 발생하였으며, 팔당호에서는 6월과 7월에만 관찰되었다. 분자생물학적 방법(TOX2P/TOX2F primer set)을 이용한 실험에서 팔당호를 제외한 세 개의 호수(영천호, 안계호, 가창호)에서 모두 microcystin 농도가 검출되지 않은 남조류 발생초기인 4월과 5월에 mcy 유전자가 확인되었다. HPLC법을 이용한 microcystin의 정성ㆍ정량적 분석에서 영천호는 microcystin(-RR, -YR, -LR)이 6월 ~ 12월(9.5 μgㆍg-1 ~ 821.1 μgㆍg-1), 안계호는 6월 ~ 11월(40.3 μgㆍg-1 ~ 356.3 μgㆍg-1), 가창호는 6월 ~ 11월(44.6 ~ 169.3 μgㆍg-1)에 microcystin이 각각 검출되었으며, 이들 상수원지에서 모두 공통적으로 microcystin-RR과 -LR의 검출빈도가 높았으며, 특히 microcystin-RR의 농도가 가장 높았다. 영천호, 안계호 및 가창호에서 M. aeruginosa는 microcystin 농도와 가장 높은 상관성을 나타내는 주요 독성종으로 확인되었으며, 총인과 총질소 등의 영양염이 M. aeruginosa의 증식을 유발함으로써 간접적으로 microcystin 생성의 증가에 영향을 주는 것으로 나타났다. 현장에서 분리한 M. aeruginosa 균주들의 microcystin 생성비율(75%)은 다른 Microcystis 균주들 보다 높았으며, microcystin 정성·정량적 분석결과 또한 현장의 microcystin 생성패턴과 매우 유사하였다. 높은 농도의 microcystin 생성이 관찰된 M. aeruginosa YC, M. ichthyoblabe AK와 M. viridis AK는 배양실험 조건에서 최적의 성장 온도(30 ~ 35℃)보다 낮은 20℃와 10 ~ 20 mgㆍL-1의 질소 농도에서 가장 높은 microcystin 생성을 나타내었다. M. aeruginosa YC는 120 μmolㆍm-2ㆍs-1의 조도와 0.5 mgㆍL-1의 인 농도에서 microcystin 생성의 최대값을 나타내었다. 반면에, M. ichthyoblabe AK와 M. viridis AK는 각각 70 μmolㆍm-2ㆍs-1와 35 μmolㆍm-2ㆍs-1의 조도와 5 mgㆍL-1의 인 농도에서 microcystin 생성의 최대값을 나타내었다. 우리나라의 조류예보제 기준(남조류 세포수)과 WHO (1998)의 microcystin-LR 가이드라인(1 μgㆍL-1)에 기초하여 상수원수내 microcystin의 위해성을 검토한 결과, 영천호와 안계호는 위해가능성이 있는 것으로 간주하는 조류경보 단계(5,000 cellsㆍmL-1 초과)가 각각 6개월(6월 ~ 11월)과 3개월(9월 ~ 11월)동안 지속된 반면, 가창호는 5월부터 10월까지 위해성이 없는 것으로 간주하는 조류주의보 수준(500 cellsㆍmL-1 초과, 5,000 cellsㆍmL-1 미만)을 유지하였다. 하지만, WHO (1998)가 정한 가이드라인에 기초했을 때, 가창호는 6월 ~ 11월에 최대 4배 초과하였으며, 안계호는 6월 ~ 11월에 약 1배 ~ 8배 초과하였다. 반면에, 영천호는 8월 ~ 11월 사이에 2배 ~ 33배 초과하였다. 더욱이 Falconer et al. (1994)이 제시한 세포당 microcystin 농도 계산식을 적용한 가창호(M. aeruginosa A3, M. aeruginosa JE10)와 영천호(M. aeruginosa SE20) 균주들의 실험결과들은 우리나라의 조류경보수준(5,000 cellsㆍmL-1)의 1/2 이하인 약 2,000 cellsㆍmL-1에서 독소 가이드라인인 1 μgㆍL-1의 microcystin 농도와 일치하여 M. aeruginosa를 중심으로 조류경보수준의 재검토가 필요한 것으로 조사되었다. 영천호에서 채집한 어류들의 체내 축적된 microcystin이 인체에 미치는 잠재적 위해성을 알아보았다. TDI (total daily intake) 값과 비교했을 때 어류들은 모두 간에서 EDI (estimated daily intake) 값이 TDI 값을 모두 초과하였으며, 떡붕어의 간에서 EDI 값이 가장 높게 나타났고 TDI 값의 약 8배 초과하였다. 따라서, 영천호에서 남조류 대발생시 떡붕어 섭취에 의한 위해가능성이 확인되었다.
This study examined the production characteristics of hepatotoxic microcystin, produced by cyanobacteria living in Lake Yeongchun, Ankei, Gachang and Paldang, to identify possibility of health risks from microcystin. It also aims to suggest a guideline for water quality based on risk assessment by a...
This study examined the production characteristics of hepatotoxic microcystin, produced by cyanobacteria living in Lake Yeongchun, Ankei, Gachang and Paldang, to identify possibility of health risks from microcystin. It also aims to suggest a guideline for water quality based on risk assessment by analyzing the production level of microcystin per cell, produced by Microcystis. On-site research on the average concentration of total phosphorus, detected in the source water, demonstrated that all the subject lakes were in above the mesotrophic status. The water of Lake Paldang contained the richest nutrients, followed by Lake Ankei, Yeongchun and Gachang. In Lake Yeongchun, four major strains of cyanobacteria, Microcystis aeruginosa, M. ichthyoblabe, M. flos-aquae and A. spiroides, were observed. M. aeruginosa, M. ichthyoblabe, M. flos-aquae, M. wesenbergii and A. spiroides were surveyed in Lake Ankei. Lake Gachang only contained M. aeruginosa, and M. aeruginosa, M. flos-aquae, M. novacekii, M. wesenbergii and Anabaena sp. were found in Lake Paldang. Standing crops of cyanobacteria invetigated in Lake Yeongchun were from 250 to 67,200 cellsㆍmL-1 during April to December. During April to November in Lake Ankei and Gachang, from 10 to 228,510 cellsㆍmL-1 and from 33 to 6,000 cellsㆍmL-1 of cyanobacteria appeared respectively. The amount of cyanobacteria observed in Lake Paldang in June and July was from 532 to 4,200 cellsㆍmL-1. The average appearance and standing crops of cyanobacteria increased in proportion to the average concentration of total phosphorus in all the subject lakes but Paldang. A test performed to analyze quantity and quality of microcystin in the source water with HPLC method found a minimum level of 9.5 ugㆍg-1 and a maximum level of 821.1 ugㆍg-1 microcystin (-RR, -YR, -LR) in July and October, respectively, in Lake Yeongchun during June to December. Water of Lake Ankei contained microcystin during June to November, a minimum of 40.3 ugㆍg-1 in July and a maximum of 356.3 ugㆍg-1 in October. From 44.6 to 169.3 ugㆍg-1 of microcystin was detected during June to November in Lake Gachang, showing high level concentration during June to September. The level of microcystin was the most high in Lake Yeongchun, followed by Ankei and Gachang. Microcystin-RR and –LR were detected frequently in all the three sources, especially microcystin-RR at the highest concentration. In another test conducted with molecular biological method using TOX2P/TOX2F primers, mcy gene, producing microcystin, was identified in all the three lakes during the study period. The gene was also found in April and May, early stage of appearance of cyanobacteria in which no microcystin was detected even by HPLC analysis. This result demonstrates the molecular biological method suggested in this study is appropriate for early detection of potential toxic cyanobacteria ahead of the period when the examination of microcystin is available by analysis of concentration. Analysis on the relationship between community of cyanobacteria and microcystin production in the three lakes, Yeongchun, Ankei, and Gachang presented that M. aeruginosa was closely related to the production of total microcystin (R2=0.788, p<0.0001), microcystin-RR (R2=0.724, p<0.0001) and microcystin-LR (R2=0.890, p<0.0001), and was the most highly related to the most toxic microcystin-LR. In addition, nutrients such as total phosphorus and total nitrogen appeared to cause growth of M. aeruginosa, thereby having an indirect effect on increase of microcystin. According to the result of PCR test for 134 strains of Microcystis, mcy gene was detected in 55 strains of M. aeruginosa and 10 strains of M. ichthyoblabe. Other species such as M. flos-aquae, M. wesenbergii and M. novacekii contained low or no gene. The result of culture of M. aeruginosa, M. ichthyoblabe and M. viridis, producing microcystin among the isolated strains, indicated that all strains presented maximum production of microcystin at 20℃, lower than optimal growth temperature of 30~35℃, and at nitrogen concentration of 10~20 mgㆍL-1. The production was also affected by limited phosphorus concentration of 0.1~0.5 mgㆍL-1. Fish in Lake Yeongchun was analyzed by species and parts to assess and estimate potential human risk of microcystin. Comparing with the total daily intake (TDI) of microcystin, the estimated daily intake (EDI) level of microcystin in muscle and gill of fish in Yeongchun Lake did not exceed the TDI level, but EDI in liver exceeded TDI two to eight times. In particular, EDI in liver of Carassius cuvieri marked the highest level. The result of the test, analysis of potential health risk of microcystin through species of cyanobacteria and concentration of mycrocystin in water supply lakes presented that Lake Yeongchun and Ankei kept cyanobacteria warning level, above 5,000 cellsㆍmL-1 indicating danger of health risk, for certain period. Lake Gachang maintained the level lower than 5,000 cellsㆍmL-1, regarded as low risk to human health, during June to November. However, compared with WHO guideline (1998) for microcystin-LR, which is 1 ugㆍL-1, Lake Gachang exceeded the guideline maximum 4 times, Ankei 1~18 times and Yeongchun 2~33 times. In addition, the result of calculation for microcystin concentration per cell by Falconer et al. (1994), performed on M. aeruginosa A3 and M. aeruginosa JE10 isolated from Lake Gachang and M. aeruginosa SE20 from Lake Yeongchun, presented that 1 ugㆍL-1 of microcystin was identified at 2,000 cellsㆍmL-1, which was more than twice lower than warning level of 5,000 cellsㆍmL-1, implying high potential human health risk from microcystin in the sources of drinking water in Korea.
This study examined the production characteristics of hepatotoxic microcystin, produced by cyanobacteria living in Lake Yeongchun, Ankei, Gachang and Paldang, to identify possibility of health risks from microcystin. It also aims to suggest a guideline for water quality based on risk assessment by analyzing the production level of microcystin per cell, produced by Microcystis. On-site research on the average concentration of total phosphorus, detected in the source water, demonstrated that all the subject lakes were in above the mesotrophic status. The water of Lake Paldang contained the richest nutrients, followed by Lake Ankei, Yeongchun and Gachang. In Lake Yeongchun, four major strains of cyanobacteria, Microcystis aeruginosa, M. ichthyoblabe, M. flos-aquae and A. spiroides, were observed. M. aeruginosa, M. ichthyoblabe, M. flos-aquae, M. wesenbergii and A. spiroides were surveyed in Lake Ankei. Lake Gachang only contained M. aeruginosa, and M. aeruginosa, M. flos-aquae, M. novacekii, M. wesenbergii and Anabaena sp. were found in Lake Paldang. Standing crops of cyanobacteria invetigated in Lake Yeongchun were from 250 to 67,200 cellsㆍmL-1 during April to December. During April to November in Lake Ankei and Gachang, from 10 to 228,510 cellsㆍmL-1 and from 33 to 6,000 cellsㆍmL-1 of cyanobacteria appeared respectively. The amount of cyanobacteria observed in Lake Paldang in June and July was from 532 to 4,200 cellsㆍmL-1. The average appearance and standing crops of cyanobacteria increased in proportion to the average concentration of total phosphorus in all the subject lakes but Paldang. A test performed to analyze quantity and quality of microcystin in the source water with HPLC method found a minimum level of 9.5 ugㆍg-1 and a maximum level of 821.1 ugㆍg-1 microcystin (-RR, -YR, -LR) in July and October, respectively, in Lake Yeongchun during June to December. Water of Lake Ankei contained microcystin during June to November, a minimum of 40.3 ugㆍg-1 in July and a maximum of 356.3 ugㆍg-1 in October. From 44.6 to 169.3 ugㆍg-1 of microcystin was detected during June to November in Lake Gachang, showing high level concentration during June to September. The level of microcystin was the most high in Lake Yeongchun, followed by Ankei and Gachang. Microcystin-RR and –LR were detected frequently in all the three sources, especially microcystin-RR at the highest concentration. In another test conducted with molecular biological method using TOX2P/TOX2F primers, mcy gene, producing microcystin, was identified in all the three lakes during the study period. The gene was also found in April and May, early stage of appearance of cyanobacteria in which no microcystin was detected even by HPLC analysis. This result demonstrates the molecular biological method suggested in this study is appropriate for early detection of potential toxic cyanobacteria ahead of the period when the examination of microcystin is available by analysis of concentration. Analysis on the relationship between community of cyanobacteria and microcystin production in the three lakes, Yeongchun, Ankei, and Gachang presented that M. aeruginosa was closely related to the production of total microcystin (R2=0.788, p<0.0001), microcystin-RR (R2=0.724, p<0.0001) and microcystin-LR (R2=0.890, p<0.0001), and was the most highly related to the most toxic microcystin-LR. In addition, nutrients such as total phosphorus and total nitrogen appeared to cause growth of M. aeruginosa, thereby having an indirect effect on increase of microcystin. According to the result of PCR test for 134 strains of Microcystis, mcy gene was detected in 55 strains of M. aeruginosa and 10 strains of M. ichthyoblabe. Other species such as M. flos-aquae, M. wesenbergii and M. novacekii contained low or no gene. The result of culture of M. aeruginosa, M. ichthyoblabe and M. viridis, producing microcystin among the isolated strains, indicated that all strains presented maximum production of microcystin at 20℃, lower than optimal growth temperature of 30~35℃, and at nitrogen concentration of 10~20 mgㆍL-1. The production was also affected by limited phosphorus concentration of 0.1~0.5 mgㆍL-1. Fish in Lake Yeongchun was analyzed by species and parts to assess and estimate potential human risk of microcystin. Comparing with the total daily intake (TDI) of microcystin, the estimated daily intake (EDI) level of microcystin in muscle and gill of fish in Yeongchun Lake did not exceed the TDI level, but EDI in liver exceeded TDI two to eight times. In particular, EDI in liver of Carassius cuvieri marked the highest level. The result of the test, analysis of potential health risk of microcystin through species of cyanobacteria and concentration of mycrocystin in water supply lakes presented that Lake Yeongchun and Ankei kept cyanobacteria warning level, above 5,000 cellsㆍmL-1 indicating danger of health risk, for certain period. Lake Gachang maintained the level lower than 5,000 cellsㆍmL-1, regarded as low risk to human health, during June to November. However, compared with WHO guideline (1998) for microcystin-LR, which is 1 ugㆍL-1, Lake Gachang exceeded the guideline maximum 4 times, Ankei 1~18 times and Yeongchun 2~33 times. In addition, the result of calculation for microcystin concentration per cell by Falconer et al. (1994), performed on M. aeruginosa A3 and M. aeruginosa JE10 isolated from Lake Gachang and M. aeruginosa SE20 from Lake Yeongchun, presented that 1 ugㆍL-1 of microcystin was identified at 2,000 cellsㆍmL-1, which was more than twice lower than warning level of 5,000 cellsㆍmL-1, implying high potential human health risk from microcystin in the sources of drinking water in Korea.
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