깔따구 (Diptera: Chironomidae)는 저서성 대형무척추동 물로 환경오염 및 수질 모니터링에 이용되는 중요한 지표생물이다. 본 연구에서는 인천 수돗물 정수장에서 발견된 깔따구류의 정밀한 종 동정을 위해 형태적 분류와 미토콘드리아 DNA에서 cytochrome c oxidase subunit I (COI) 유전자의 염기서열을 이용하여 분석하였다. 정수장 6곳의 20 개체는 안개무늬날개깔따구 (Chironomus kiiensis) 12개체, 노랑털깔따구 (Chironomus flaviplumus) 6개체, 등깔따구 (Chironomus dorsalis) 1개체, 용산무늬깔따구 (Polypedilum yongsanensis) 1개체 등 4종으로 확인되었다. 각 깔따구 종의 형태적 특징은 두부, 하순기절, 대악, 안테나, 발톱의 형 태적 특징을 살펴보았다. NCBI Genbank에 등록된 깔따구 17종 21개체의 COI 염기서열을 바탕으로 본 연구에서 조사된 20개체의 계통진화적 분석한 결과 각 4종의 깔따구 COI 염기서열은 등록된 동인 종과 높은 상동성을 보이며 (99~100%) 같은 계통군 (clade)으로 나타났다. 이러한 결과는 국내 깔따구의 종 동정을 위한 형태적 - 유전적 정보를 통합적으로 제공함으로 담수생태계의 모니터링을 위한 주요한 정보로 활용될 것이다.

중금속은 다양한 경로를 통해 환경 중 배출되어 서식 생물에 노출되며 체내 다양한 생리학적 불균형을 유도한다. 본 연구에서는 수서생물지표종으로 이용되는 깔따구 (Chironomus riparius)를 이용하여 야외 중금속 (Al, Aluminum; Cr, Chromium; Cu, copper; Mn, Manganese; Zn, Zinc) 농도 노출에 따른 다양한 분자발현 반응과 상관성을 분석하였다. 생물 체내 분자 반응을 관찰하기 위해 heat shock protein 40, 70, 90 (HSP40, 70, 90), cytochrome 450 (CYP450), Glutathione S-transferase (GST) and Serine-type endopeptidase (SP)를 이용하였다. 그 결과, 스트레스 분자마 커로 이용되는 HSPs 유전자들은 중금속 노출된 개체들에서 대조군보다 높은 경향을 보였으며 Cu 노출 시 가장 높은 발현을 나타냈다. 해독에 관여하는 CYP450과 GST 유전자 발현 결과, Cr과 Cu에서는 다른 노출군에 비해 높은 발현 경향을 나타냈다. SP 유전자 발현 결과 Al을 제외한 모든 노출군이 대조군과 유사한 발현 패턴을 보였다. 이와 같은 연구 결과는 실내에서 환경 중 존재하는 실제 농도를 반영한 독성실험을 통해 노출물질과 농도에 따라 특이적으로 발현하는 분자마커 패턴을 보고하였다. 또한, 수생태계로 유입되는 중금속이 하천에 서식하는 생물에 주는 유해 영향에 대한 정보와 분자 지표 유전자들의 현장 적용 가능성을 보여준다.

인간의 인위적 활동으로 인해 발생하는 중금속 중 카드 뮴 (Cd)과 아연 (Zn)은 다양한 경로를 통해 하천으로 유입 되어 서식생물에게 유해 영향을 준다. 본 연구에서는 도심 하천에서 우점하는 장수깔따구 (Chironomus plumosus)를 이용하여 Cd와 Zn 노출에 따른 생태독성학적 반응을 연구 하였다. 생활사를 고려하여, 생존율, 성장율, 우화율, 성비와 하순기절 기형을 관찰하였다. 장수깔따구 생존율은 Cd 와 Zn 노출에 따라 시간, 농도의존적인 경향을 나타냈다. 성장율은 Cd 노출 후 Day 4와 Day 7, 농도의존적인 감소를 보였으며, 고농도인 50 mg L-1에서 대조군에 비해 급격한 감소를 나타냈다. 또한, 우화율은 10, 50 mg L-1 Cd에서 대조군에 비해 감소함을 관찰하였다. 성비 변화는 100 mg L-1 Zn에서 암컷의 비율이 증가였으나 농도가 높아짐에 따라 수컷의 비율이 높아짐을 보이며 성비불균형이 관찰되었다. 게다가, 장수깔따구의 하순기절에서는 대조군에 비해 Cd와 Zn 노출된 장수깔따구에서 기형이 관찰되었다. 이와 같은 결과는 수생태계로 유입되는 중금속이 하천의 하상저층에 서식하는 저서무척추동물에게 유해한 영향을 주며, 중금속 노출에 따른 생물학적 분석을 위한 현장 지표 종으로서의 가능성을 보여주었다.

Heavy metals resulted from the increase of human industrial activity are introduced into the environment through rainfall and wastewater, and have harmful effects on inhabitants. In this study, we investigated biological responses such as survival rate, growth rate, emergence rate and sex ratio, and morphological effects of mentum deformity in Chironomus plumosus, an indicator organism to evaluate pollutions on aquatic ecosystem. The survival rate of C. plumosus showed time- and dose-dependent decrease after chromium and copper exposures. Growth rate decreased at 4th day after chromium exposure and significantly reduced at exposure to relatively high concentration (copper 1000 mg L-1) for all exposure times. In addition, we observed that the emergence rate by exposure to copper 1000 mg L-1 was significantly lower than that of the control group. The imbalance of sex ratios showed at relatively low concentrations (chromium 10 and 50 mg L-1) with the high proportion of female and at the relative high concentration (copper 1000 mg L-1) with the high proportion of male. Furthermore, the morphological mentum deformities of C. plumosus observed in the exposed group according to chromium and copper exposure. These results suggest that the heavy metal exposure in environment may influence biosynthetic and morphological stresses of benthic invertebrate C. plumosus, and aquatic midge C. plumosus are potential indicators for toxicity assessment of heavy metals such as chromium and copper.

We investigate that the impact of freshwater organism exposed to the salinity environment by the frequent rainfall following climate change. To evaluate the stress response following salinity exposure, we assessed the survival rate, molting success rate, the developmental period and mouthpart deformities in Chironomus riparius. In addition, we measured the molecular responses of biomarker gene, gene expression of heat shock protein 70 (HSP70) in C. riparius exposed to salinity after 96 hour. The C. riparius survival rates were showed on time dependent manner and not observed survival organisms above 15 psu at day 4. The pupation and emergence of C. riparius were not seen above 15 psu, and the molting success rate was less than 20% at 10 psu. The developmental retardation of C. riparius was well observed in the pupation and emergence period and was delayed by 4 days at 10 psu compared to the control and 5 psu. The mouthpart deformities after salinity exposure at 96 or 72 hour were observed at 10 psu and 15 psu. The expression of C. riparius HSP70 level was significantly increased exposure to 5 psu and 10 psu. Thus, salinity has been caused to be various ecotoxicological and molecular stress responses on freshwater organisms similar to harmful substances such as EDCs and so on.

Despite the importance of behavioral ecotoxicological tests, few researches have been conducted to assess the quality of freshwater using behavior of test species, in part due to lack of standardized test method for quantifying the response. In this study, we developed a new test methods using non-biting midge larva’s burrowing behavior to assess field water quality and a new index, AUC (area under curve), to compare burrowing behavior quantitatively. Four kinds of field samples (residential, natural, agricultural and industrial areas) were tested using the method to validate our methodology and index. There were not significant differences in 96 h mortality and heavy metal concentration between samples, but AUC from the industry sample was significantly decreased.

Chironomous is aquatic insect belonging to order Diptera, family Chironomidae. Their larval stage can be found mainly in aquatic benthic environment, hence good model organism to study environmental toxicology assessments and consider as useful bio indicators of contamination of the aquatic environment. In this study, Chironomus Heat Shock Proteins, Cytochrome 450, Glutathione S-transferase, Serine-type endopeptidase gene expressions were compared between polluted field areas (Chironomus plumosus) and under laboratory conditions (Chironomus riparious) to investigate molecular indicators for environmental contaminant stress assessment. Heavy metal (Al, Fe, Mn, Cu, Cr, Zn, Se, Pb, As, Cd) concentrations in sediments collected from three study areas exceeded the reference values. Moreover, HSPs, CYP450 and GST gene expression except SP for C. plumosus showed higher expression than C. riparious gene expression. Similar gene expression pattern was observed in C. riparious that exposed environment waters up to 96 h when compared to C. plumosus exposed to waters that grown in lab conditions. In summary, this comparative gene expression analysis in Chironomous between field and laboratory condition gave useful information to select candidate molecular indicators in heavy metal contaminations in the environment.

Distribution of aquatic organisms is governed by various environmental factors Hydromorphological characteristics which are heavily affected by heavy rains in stream are one of the fundamental factors to influence the distribution of aquatic organisms. In this study we evaluated the difference response of two aquatic species Chironomus yoshimatisui and Limnodrilus hoffmesteri habiting to changes of discharge due to heavy rains. We compared the differences in their resilience and recover to disturbances. Our results showed that after a heavy rain during the sampling periods, the proportion of macrosubstrate highly increased and was not recovered into the previous states. Nonmetric Multi-dimensional scaling based on Chironomidae and Oligochaeta biota well represented annual and seasonal differences with heavy rain effects. Random forest model showed that the abundance of C. yoshimatisui were mainly influenced by microsubstrate, precipitation and Reynolds number, whereas stream roughness, mesosubstrate and macrosubstrate much more influenced the abundance of L. hoffmesteri.

Chironomus riparius, a non-biting midge (Chironomidae, Diptera), is extensively used in aquatic ecotoxicological studies for assessing acute and sub-lethal toxicities of contaminated sediments and for water monitoring due to their widespread occurrence, short life-cycle, easy to be reared in the laboratory, physiological tolerance to various environmental conditions. To date, the endpoints used for monitoring such effects in C. riparius are based on a small number of specific biomarkers and measurements of organism level effects, such as survival and reproduction. Genomic-based techniques based on expression analysis of genes are important tools for investigating molecular level effects caused by exposure to environmental pollutants, which will provide the ability to detect mechanisms of action and subsequent adverse cellular level effects and associated with different types of toxicity. As a pre-requisite for genomic based ecotoxicological studies knowledge on the C. riparius transcriptome is important but despite its ecotoxicological importance, no large scale transcriptome analysis of C. riparius has been done so far. Therefore, to gain a better understanding of C. riparius transcriptome, we recently developed Expressed Sequence Tags (ESTs) sequencing project on C. riparius larvae using 454 pyrosequencing. Sequencing runs, using normalized cDNA collections from fourth instar larvae, yielded 20,020 expressed sequence tags, which were assembled into 8,565 contigs and 11,455 singletons. Sequence analysis was performed by BlastX search against the National Center for Biotechnology Information (NCBI) nucleotide (nr) and uniprot protein database. Based on the gene ontology classifications, 24% (E-value ≤1-5) of the sequences had known gene functions, 24% had unknown functions and 52% of sequences did not match any known sequences in the existing database. Sequence comparison revealed 81% of the genes have homologous genes among other insects belonging to the order Diptera providing tools for comparative genome analyses. Targeted searches using these annotations identified genes associated with essential metabolic pathways, signaling pathways, detoxification of toxic metabolites and stress response genes of ecotoxicological interest. The results obtained from this study would eventually make ecotoxicogenomics possible in a truly environmentally relevant species, C. riparius. Various C. riparius ecotoxicity studies using stress response genes developed from 454 sequencing will be presented in the conference.

농업생태계의 환경별로 깔따구류의 분포를 밝히고 우점종을 이용한 유기물 분해능을 평가하고자 대단위경작지, 친환경농업지, 생활하수 유입지, 축산폐수 유입지 및 주변수로 등의 5개 서식처별로 나누어 '04~'06년까지 각 구획을 대표하는 총 78개 지점에서 깔따구과 및 공서 무척추동물을 정량 채집하였다. 그 결과, 농업지대의 깔따구류는 총 16종으로 동정되었다. 서식지 유형별로 출현한 종수를 보면, 축산폐수 유입지에서 11종, 대단위 농경지는 8종,

Acetaminophen (CAS 103-90-2) is one of the most used pharmaceuticals around the world. In Korea, it was produced 1,069 tons in 2003. This chemical is not eliminated in wastewater treatment plant and may flow into the ecosystem through various routes. Therefore, there is a possibility that it can make an adverse effect on aquatic organisms. To examine its ecological toxicity, we used three native Korean aquatic invertebrate species, Daphnia sp., Chironomus yoshimatsui, and Ephemera orientalis. The acute toxicity on Daphnia sp. was moderately high, and its 48 hour median immobilization concentration (EC50-immobilization) was 51.7 mg/L. On the other side, the reproductive toxicity was very high, and its EC50 of 25 day reproduction test was 0.005 mg/L. In E. orientalis egg hatching test, the median egg hatching inhibition concentration was 0.199 mg/L. C. yoshimatsui was most tolerant to acetaminophen, in which 48 hour median lethal concentration (LC50) was 400.0 mg/L and 45 day median emergence inhibition concentration (EC50-emergence) was 45.27 mg/L. From this results, we concluded that acetaminophen is hazardous to freshwater macroinvertebrates, especially to water flea. Therefore we need to study more about pharmaceuticals' ecotoxicology including acetaminophen and to assess their potential ecological risk.

Traditionally oxygen demands and concentrations of variable pollutant have been used for the assessment of lentic or lotic water quality. But there is a high probability that newly synthesized chemicals flow into the aquatic ecosystem, and it is very hard to investigate that many kinds of chemicals interact even though they exit at lower level than legal standards. As a consequence of limitation of chemical assessment for media, biological assessment techniques for accepters which are affected by pollutants or physicochemical properties have been developed. However, it still has several weak points because it does not have the ecological relevances using the internationally standard test species which do not inhabit Korea. Besides, the assessment using just one or two species may have a possibility of overestimation or underestimation because a certain species can be extremely sensitive or insensitive to a certain chemical or physicochemical property. Therefore, we used four kinds of indigenous aquatic invertebrates to assess the local stream quality and integrated their responses into a new index which ranges from 0(bad) to 1(good). We assessed four streams throughout Ansan and one stream in Gapyeong, and index values of two streams near residential and industrial area in Ansan were under 0.9 and the others were over 0.9.

실내에서 사육된 C. riparius를 대상으로 내분비계 교란물질인 DEHP를 처리하여 노출시간에 따른 기형발생률과 발생한 기형의 유형을 세분화하였다. 하순기절 구조별 기형발생률은 MLT (medial lateral tooth)에서 가장 높았고 그 다음이 LT (lateral tooth), MIX 순으로 높게 나타났다. 비처리군과 처리군 모두 MLT 단독의 기형이 가장 많았다. 하순기절의 기형유형은 여섯 가지로 세분화하였다. 발생빈도가 높은 기형유형은

The exposed strain of C. riparius treated with di (ethyl- hexyl)-phthalate (DEHP) did not result in a consistent relationship between mortality or sex ratio and chemical concentrations. And after treating with DEHP, the emergent female from the exposed st

Development of the fourth-instar larvae of Chironomus riparius has a sensitive to ecdysteroid hormones. The 2D/E gel analysis for polypeptide expression reflecting early-ecdysterone inducible gene has conducted the emerged female from larval phase exposur

The 2D/E gel analysis for polypeptide expression reflecting I-18 C gene (early-ecdysterone inducible gene) has conducted the emerged C. riparius adults from larval phase exposure to tebufenozide acting as an ecdysteroidal molting hormone. Control group, t

한국산 요시마쯔깔따구(Chironomus yoshimatsui)의 세 가지 중금속(납, 카드뮴, 수은)에 대한 급성독성 및 행동 독성을 외국 표준 실험 종인 리파리깔따구(C. riparius)와 비교하였다 48시간 및 96시간 동안 수중 노출(water-only exposure)을 시킨 후 두 가지 실험종의 반수치사농도(LC50)및 깔따구의 유영 능력 감소에 영향을 주는 농도(EC50)를 바탕으로 두 실험 종간의 차이를 살펴보았다. 카드뮴