초록 ▼

1. AFS 처리활성물질 분석기술 개발
1-1. AFS 처리활성물질 분석기술 정립 및 공정시험기준안 도출 : CuPT, ZnPT, Diuron, Irgarol, Sea-Nine에 대한 질량분석법을 정립하고 이를 바탕으로 시범지침서를 작성함.
1-2. 선박부착 및 수중제거 파생물 시험적용 : 파생물 내 중금속 및 유기계살생물질 잔존 특성을 파악하고 배출되는 입자성 물질의 크기별 분포특성을 파악함.

2. 선박부착 파생물의 LAB-SCALE영향평가, 지표발굴, 모사실험 및 AFS활성물질 영향 평가
2-1. 선

1. AFS 처리활성물질 분석기술 개발
1-1. AFS 처리활성물질 분석기술 정립 및 공정시험기준안 도출 : CuPT, ZnPT, Diuron, Irgarol, Sea-Nine에 대한 질량분석법을 정립하고 이를 바탕으로 시범지침서를 작성함.
1-2. 선박부착 및 수중제거 파생물 시험적용 : 파생물 내 중금속 및 유기계살생물질 잔존 특성을 파악하고 배출되는 입자성 물질의 크기별 분포특성을 파악함.

2. 선박부착 파생물의 LAB-SCALE영향평가, 지표발굴, 모사실험 및 AFS활성물질 영향 평가
2-1. 선박부착 파생물의 LAB-SCALE영향 및 지표발굴
2-1-1. 넙치: 고압세척용출수(Water jet effluents)와 MeOH 추출물에 노출된 배아는 공통적으로 세포 형태형성, 신호전달, 근절섬유 분절기작에 유의하게 영향을 받음
2-1-2. 곤쟁이/단각류: 고압세척 용출수는 중금속독성영향으로 인해 MeOH 추출액보다 발생독성이 4배에서 7배까지 높음
2-1-3. 둥근성게: 이어도호는 고압세척용출수의 독성이 강한 것으로 나타났고, 이사부호는 MeOH 추출액이 고압세척용출수에 비해 독성이 강한 것으로 나타났음.
2-1-4. 동물플랑크톤: 대표 연안요각류 중 Paracalanus parvus s.l 이 Acartia omorii보다 독성에 더 민감함

2-2. 선박부착생물 모사실험 및 LAB-SCALE 영향 평가
2-2-1. 넙치: 심장부종 빈도는 Biocide, Silicone, background 실험구의 10,000배 희석구간에서만 유의한 증가를 확인
2-2-2. 둥근성게: 모사판 실험에 의한 고압세척용출수 내 독성은 biocide 처리구에서 가장 높았고, MeOH 추출액 중에서는 background 처리구의 독성이 가장 강하게 나타났음.
2-2-3. 동물플랑크톤: Silicone 도포 모사판 시험구에서 독성이 가장 낮았고, 나머지 시험구들은 유의한 독성 차이가 없었음

2-3. 국내 주상용 Principal biocide AFS처리 활성물질에 대한 독성영향평가
2-3-1. 넙치: 3 종(Sea-Nine, Irgarol, Diuron)의 booster biocides에 배아가 노출 되었을 때, 치골형성, 신호전달, 중추신경뉴런발달 기능 유전자 등의 기작이 유의한 영향을 받음
2-3-2. 곤쟁이/단각류: 곤쟁이는 단각류보다 3배 가량 민감한 반수치사율을 보임
2-3-3. 둥근성게: Irgarol의 EC₅₀은 4,476 ㎍/L, Diuron의 EC₅₀은 10,777 ㎍/L, Sea-Nine의 EC₅₀은 8.134 ㎍/L CuPT의 EC₅₀은 3.423 ㎍/L, ZnPT의 EC₅₀은 6.488 ㎍/L였음. 둥근성게 배아에 미치는 독성은 CuPT가 가장 높은 것으로 나타났음.
2-3-4. 동물플랑크톤: Sea-Nine 211이 다른 활성물질에 비해 매우 강한 독성을 나타냄

3. 선박부착기인 생물 및 위해성 요인 탐색 및 판별시도
3-1. 선저부착생물 확보 및 판별 그리고 위해성요인 탐색
3-1-1. 미세조류: 선박 방오도료 내성 미세조류 탐색 (Olfantiella onnuriae)
3-1-2. 원생생물: 4척의 선박에서 섬모충 6종 탐색, 장목 2호에서 분리된 Euplotes 2종은 국내 미기록 종으로 확인됨
3-1-3. 부착대형저서동물: 5척의 연구조사선을 대상으로 부착대형저서동물을 조사한 결과 총 47종이 출현하였으며, 평균 10,297 개체/m²와 생체량 2,381 gWWt/m²이 출현함

3-2. 선박부착생물 모사실험 파생생물 판별
3-2-1. 미세조류: 선박부착생물 모사판을 통한 부착 미세조류 군집 파악 및 파생미세조류의 Viability 연구 수행
3-2-2. 원생생물: 부착판 섬모충 4종 탐색, 방오도료 처리와 미처리 판의 섬모충 초기생물상은 차이를 보임
3-2-3. 부착대형저서동물: Background 모사판에는 유령멍게(Ciona intestinalis), 거친대추멍게(Ascidiella aspersa), 큰다발이끼 벌레(Sugula neritina)가 부착하였으며 Treatment 모사판에는 주로 주걱따개비(Balanus amphtrite), 삼각따개비(Balanus trigonus)가 부착함

4. 수중제거 핵심 관리요소 선정 및 관리 framework 제시
4-1. 수중제거 시나리오 탐색: 현재 사용되는 수중제거 기술은 크게 manual removal, mechanical removal, surface treatment, shrouding techniques 방식으로 구분되며, 국가 간 이동하는 대형 상선에서는 mechanical removal, surface treatment 방식이 주로 사용됨
4-2. 관리 수준 대응 핵심 요소 도출: 위해성 평가/관리 대응 핵심요소를 기반으로 40 가지의 수중제거 시나리오를 작성함
4-3. 수중제거관리를 위한 위해성평가 기법 제시: 수중제거 시나리오에 대한 RPN 산정 및 MAMPEC 모델 구동에 의한 생물학적·화학적 위해성평가 절차를 완성하여 시범적용 평가하였음

(출처 : 초록 3p)

Abstract ▼

IV. Results
a. Development of analysis technique for active substances from antifouling system
(1) Establishment of analysis techniques and guidelines for active substances discharged from antifouling system
To establish analysis method for active substances discharged from antifouling sys

IV. Results
a. Development of analysis technique for active substances from antifouling system
(1) Establishment of analysis techniques and guidelines for active substances discharged from antifouling system
To establish analysis method for active substances discharged from antifouling systems, target analytes were first selected based on the international regulations, as well as international market share of antifouling paints. Copper pyrithione and zinc pyrithione were selected because they are most actively used in antifouling paints worldwide. Diuron, Irgarol, and DCOIT (Sea-Nine) were also included because they were highly used in the past and still persist in the environment. In addition, metallic copper and zinc were included, which are one of the main ingredients in antifouling paint as active substances. Mass library of each biocides were obtained and Q1/Q3 ions were selected from the library for quantification of biocides using liquid chromatograph and mass spectrometer. The mass spectrometric conditions and liquid chromatographic separation were further optimized for more sensitive detection of the analytes. Based on the developed methods, a pilot guideline was prepared for the standardization of the analysis. It includes introduction, consideration for sampling, sampling and preservation methods, pre-treatment methods, QA/QC, and analytical methods in annex 1, 2, and 3.

(2) Application of the analysis techniques to the wastes from haul-out or in-water cleaning activities
Zinc pyrithione (172 - 260 ppb) and copper pyrithione (28.8 - 187 ppb) were detected in all wastewater collected during haul-out cleanings. Diuron was detected only in the wastewater from JANGMOK 2. Sea—Nine and Irgarol were not detected in all samples. The concentrations of zinc pyrithione and copper pyrithione were equivalent to the lethal concentrations for the 35 to 61% of the species reported in the ECOTOX database. Concentrations of copper and zinc in the wastewater from haul-out cleaning exceeded marine water quality standard and discharge criteria by 3,400 and 22 times, respectively. They were higher than the lethal concentrations for almost all species reported in the ECOTOX database. Iron (12-25%), zinc (16-21%), and copper (0.5-5%) were major components in the particle of the waste waters. Total suspended solid (TSS) in the wastewater was 1,570 mg/L, exceeding discharge criteria to ‘clean’ area and STP discharge criteria by 80 and 319 times, respectively. Particle size distribution in the wastewater and the relationship between particle size and cumulative particle weight was also obtained. Metal compositions in seaweeds collected from ship’s hull were similar to that in the particles of wastewater, mostly consisting of zinc and copper. The concentrations in the seaweeds were up to 68 and 220 times higher than background concentrations of copper and zinc, respectively. In the paint chips and sediment collected at the shipyard, copper concentration was highest and followed by iron, zinc, and barium, which is unlike the composition in the wastewater or in seaweeds. Copper is the most frequently added component in antifouling paint. TSS in seawater was 897 mg/L on average. After the release of the wastewater into adjacent seawater, the seawater TSS was increased by 10%. Copper, zinc, manganese, and barium, the major components in antifouling paints, were also higher in adjacent seawater. The concentrations were increased by about two times after cleaning activity, exceeding marine water quality standards by 39 and 9.4 times for copper and zinc, respectively. TSS in the wastewater from the in-water cleaning simulation exceeded discharge criteria to ‘clean’ area and STP discharge criteria by 55 and 218 times, respectively. Zinc (557 ppb), copper (359 ppb), and iron (103 ppb) were relatively higher in the wastewater from the plate covered with antifouling paint. The concentrations of copper and zinc exceeded the marine water quality standards by 120 and 17 times, respectively. Booster biocides were not detected in all wastewater from simulation plates. Copper (5,170 ppb) and zinc (1,070 ppb) were also relatively higher in the particles collected in the wastewater from the plate covered with antifouling paint. The removal of foulants is expected to increase the concentrations of dissolved active substances in seawater, as well as to continuously release active compounds into the water column as a large amount of particles move along the water circulation.

b. Simulated experiment, assessment and development of index for lab-scale effects on the wastes from ship’s surface during in-water cleaning activities and active substances of antifouling systems
(1) Assessment and development of index for lab-scale effects on the wastes from ship’s surface during in-water cleaning activities
(a) Evaluation of toxicity test for embryo development of flounder Antifouling constituents enter the marine environment following the removal of spent residues during hull maintenance. Removal of antifouling paint by scraping, blasting or hosing usually takes place on the hard standings of marinas and harbours, and fine residues are readily transported into the marine environment where they become interspersed with sediments, creating ‘hotspots’ of contamination. Compared with leached biocides, those bound to particles of the paint matrix are considerably more persistent and are, therefore, likely to pose a longer-term threat to the local marine environment. The present study determined the developmental toxic effects of the high-pressure cleaning effluent (R/V EARDO and IS ABU) and methanol extract on the early developmental stages of flounder (Paralichthys olivaceus). At 48h after exposure, embryos were shown 100% of mortality in the exposure group of 100 dilution rate for high-pressure cleaning effluent (R/V EARDO) and methanol extract. Overall, two materials produced a largely overlapping suite of malformation defects, marked by the well-known effects including caudal fin fold defects, dorsal curvature, and pericardial edema. But, the embryonic flounder exposed to water jet effluent produced the higher malformation effects than those of methanol extract exposure group. We used high-throughput sequencing (RNA-seq) to characterize the developmental toxic effects from oil exposure. In gene ontology analysis, genes associated with cell morphogenesis, regulation of signal transduction, and sacomere significantly changed (cutoff P<0.01) in embryonic flounder exposed to high-pressure cleaning effluent (R/V EARDO). Genes associated immune system, reproduction and development significantly changed in emrbyonic flounder exposed to high-pressure cleaning effluent (R/V ISABU) (cutoff P <0.01).

(b) Evaluation of toxicity test for mysids & amphipods
Since antifouling is effected by the slow, controlled leaching of biocides from the painted surface, elevated environmental concentrations of these chemicals are most significant in semi-enclosed marine systems, such as harbours, marinas and estuaries, where the transport, berthing or docking of vessels is important. In most cases, previous risk assessment of these biocides has been inadequate so that their possible effects on aquatic ecosystems is a matter of great concern. The first-year study was determined the acute toxic effects of high-pressure cleaning effluent (R/V EARDO) and methanol extract on mysids and amphipod. The value of LC₅₀ for amphipod exposed to high-pressure cleaning effluent and methanol extract were 0.642% and 2.798%, respectively. The value of LC₅₀ were 0.412% and 2.680% in mysids exposed to water jet effluent and methanol extract. Regarding the toxic effects, the high-pressure cleaning effluent was more toxic than methanol extract in mysids and amphipod. The second-year of study was determined the acute toxic effects of high-pressure cleaning effluent and methanol extract from R/V ISABU on mysids and copepod. The value of LC₅₀ for copepod exposed to high-pressure cleaning effluent and methanol extract were 0.006% and 0.056%, respectively. The value of LC₅₀ were 0.232% and 0.084% in mysids exposed to high-pressure cleaning effluent and methanol extract from R/V ISABU.

(c) Evaluation of toxicity test for fertilization of sea urchin
To assess the adverse effects of the wastes on marine biota from ship’s surface during in-water cleaning activities, we conducted the toxicity test for by-products using sea urchin embryos. The fertilization rate of sea urchin embryos exposed on the water samples from the water jet operation on the R/V EARDO was 79±5.8% at 1,000-fold diluted solution, and the fertilization rates exposed in l -fold and 100-fold diluted solutions were 0.2±0.5% and 2.3±1.4%, respectively. The fertilization rate in the water jet samples from the R/V ISABU was 0.2% in 10-fold diluted solutions, but normal fertilization was found (>90%) in 100-fold diluted solution. The fertilization rate on the MeOH extracted solution of wastes from the R/V EARDO was 0% in both 30 and 300-fold diluted solutions. The fertilization rate was more than 90% in 3,000-fold diluted solutions. The fertilization rate on the MeOH extracted solution from the R/V ISABU was 0% in both 2.5 and 250-fold diluted solutions. The normal fertilization was found (>90%) in 2,500-fold diluted solutions. The highest toxicity on sea urchin embryos was found in the wastes from the water jet operation on the R/V EARDO, and the toxicity of MeOH extract was higher than wastes from the R/V ISABU.

(d) Evaluation of toxicity test for zooplankton
Effluents from ship bottom washing using a high pressure washer were used to determine the toxic effects of ships’ biofouling during the ship repair in the R/V EARDO of Korea Institute of Ocean Science and Technology Research. Experiments on ships’ biofouling have confirmed egg hatching rates and larval mortality rates of Acartia omorii and Paracalanus parvus s.l. for water-jet effluents and water-jet methanol extracts. In experiment section for each concentration, the pH was 8.04±0.03 and the saturation of dissolved oxygen(DO) was 98.38±0.37 which was almost saturated. The egg hatching rate of P. parvus s.l. in a water-jet methanol extract was slightly higher than that in a water-jet effluent, but it was difficult to compare clearly in the case of A. omorii. The larval mortality was higher a water-jet effluent than a water-jet methanol extract in both of the copepods, indicating that the toxicity of a water-jet effluent was stronger. Overall, P. parvus s.l. was more sensitive to Development of risk assessment for control technology to reduce transfer of ship’s biofouling： In-water cleaning technique toxicity than A. omorii. The LC₅₀ of P. parvus s.l. in 24h-toxicity test showed that the water-jet effluent (about 9,000 times dilution) was about 15 times higher than the water-jet methanol extract (about 650 times dilution).

(2) Simulated experiment for ship’s biofouling and lab-scale effects on the wastes from the simulated experiment
(a) Evaluation of toxicity test for embryo development of flounder
In the second-year of study, we evaluated the toxic effects on embryonic flounder exposed to waste water from in-water cleaning simulation. Four experimental group were divided including the Biocide, No-Biocide, Silicone and background (control) groups. After 70 days, they washed by sterilized-filtered sea water and diluted to 1,000, 10,000, 100,000 times using sterilized-filtered sea water. Frequency percentage of tail-fin defects and spinal curvature increased in embryonic flounder exposed to cleaning water from silicone and background groups (dilution of 10,000 times group). Frequency percentage of pericardial edema increased in embryos exposed to cleaning water from Biocide, silicone and background groups (dilution of 10,000 times group).

(b) Evaluation of toxicity test for fertilization of sea urchin
The stock solution for the sea urchin embryo test was obtained by submerging four different kinds of simulated plates treated with different biocide in sea water for two months and wastes from the water jet operation and their MeOH extracted solution. As a results of fertilization rate for water jet samples, ‘Biocide’ treated plates showed more than 90% fertilization rate from 100-fold diluted solution. In the 'No-Biocide5 and ‘Background’ treated plates, the fertilization rate was different from control in only undiluted solution, and ‘Silicone’ treated plates showed no difference from the control in the undiluted solution. In the 'No-Biocide5 treatment, the fertilization rate after filtration was higher than 90%, and it is considered to be the effect of the particulate matter rather than the toxicity in the water jet. As a results of fertilization rate on MeOH extracts of each treatment and the ‘Biocide’ treatment showed more than 80% fertilization rate from l -fold dilutions. 'No-Biocide5 treatment showed more than 90% fertilization rate from 100-fold dilution. ‘Silicone’ treatment was 96.3±2.1% in 10-fold dilution. In the ‘Background’ treatment, the fertilization rate was 0% in both 10- and 100-fold diluted solutions.

(c) Evaluation of toxicity test for zooplankton
A static panel test was conducted to evaluate the toxic effects of zooplankton on AFS effluents. (EXP. 1: Background + Tie coat, EXP. 2: Background + Tie coat + Pain, EXP. 3: Background + Tie coat + Biocide, EXP. 4: Background + Tie coat + silicone). The three hatching rates for effluent dilution concentration were from 50% to 97% in the three test groups (EXP. 1, EXP. 2, EXP. 3) that were predicted to have a toxic gradient in the hatching rate test for AFS effluent. There was no statistically significant difference. These results imply that there is a need to verify the concentration of the antifouling paint active material and further experiments. These results should be verified through an analysis of the active substance concentration of the eluted simulated antifouling paints in the test water and additional toxicity tests. The test group of EXP. 4 shows a hatching rate of over 90% at all concentrations, indicating that there is no toxic effect in the effluent of a static panel test.

(3) Assessment of active substances for principally used in domestic antifouling system
(a) Evaluation of toxicity test for embryo development of flounder
The use of alternative biocides for antifouling application has increased since the restriction on the use of organotin compounds. However, there is the limited information of those biocides on the developmental toxicity to non-target marine organism. Assessment of the toxic effects associated with alternative biocides is also needed on resident species. The present study determined the developmental toxic effects of the alternative antifouling compounds including Diuron, Irgarol 1051, Sea-Nine 211, CuPT, ZnPT on the early developmental stages of flounder (Paralichthys olivacevs). At 48h after exposure, frequency percentage of mortality was < 10% in all the exposure group of Irgarol 1051 and Diuron. But embryos were shown 100% of mortality in the exposure group of 100 ㎍/L for Sea-Nine 211 and 1,000 ㎍/L for CuPT. Overall, five biocides produced a largely overlapping suite of defects, marked by the well-known effects including caudal fin fold defects, dorsal curvature, and pericardial edema. Those biocides may be ranked in the following order from highest malformation and mortality； Sea-Nine 211 > Irgarol 1051 〉Diuron. Embryos exposed to CuPT were shown the higher frequency percentage of malformation than those of ZnPT. We used high-throughput sequencing (RNA-seq) to characterize the developmental toxicity of the five biocides. Embryos exposed to Diuron showed changes related to cellular protein localization, whereas genes associated with immune system processes were up-regulated significantly in embryos exposed to Irgarol 1051®. Genes related to actin filament organization and embryonic morphogenesis were up-regulated in embryos exposed to Sea-Nine 211®. The genes related to metabolism, immune, development were variated significantly. The genes associated with cancer immune, development were changed in embryos exposed to ZnPT, significantly.

(b) Evaluation of toxicity test for mysids & amphipods
After the definitive ban on tin-based antifouling substances, new organic compounds have recently been introduced in antifouling paint formulations, as either principal or booster biocides. In most cases, previous risk assessment of these biocides has been inadequate so that their possible effects on aquatic ecosystems is a matter of great concern. We studied the acute toxic effects of new organic biocides often associated in paint formulations, Sea-Nine 211 on mysids and amphipod. In the study, The LC₅₀ for amphipod and mysids exposed to Sea-Nine 211 were 18.3 ㎍/L and 6.09 ㎍/L. Reports regarding the toxic sensitivity from Sea-Nine 211 exposure, the mysid was more sensitivity than those of amphipod.
In second-year study, we calculated the HC₅ value (Hazardous concentration 5% based on SSD graph (Species Sensitivity Distribution graph) using the toxic data base. In results, fish tend to be more sensitive than crustaceans. The value of LC₅₀ were 69.56 ㎍/L and 302.69 ㎍/L in biota exposed to CuPT and ZnPT and the value of HC₅ were 0.125 ㎍/L and 0.52 ㎍/L, respectively.

(c) Evaluation of toxicity test for fertilization of sea urchin
As a result of the fertilization rate of sea urchin for 5 kinds of biocide, the value of EC₅₀ of Sea-nine was 8.134 ㎍/L, but those of Diuron and Irgarol were 10,777 ㎍/L and 4,476 ㎍/L, respectively. The value of EC₅₀ of CuPT and ZnPT were 3.423 ㎍/L and 6.488 ㎍/L, respectively. Among 5 AFS active substances, the highest toxicity was found in CuPT.

(d) Evaluation of toxicity test for zooplankton
Experiments on ship’s biofouling have confirmed egg hatching rates and larval mortality rates of Acartia omorii and Paracalanus parvus s.1. for active substances from antifouling system, such as Diurom, lrgarol 1501, Sea-Nine 211, CuPT, and Zn PT. Test organisms, which is commonly dominant in the coastal area, were directly collected in the situ. In experiment section for each concentration, the pH was 8.07±0.02 and the saturation of dissolved oxygen(DO) was 98.30±0.26 that was almost saturated. The eggs hatching rates of the two species of copepod were higher in order of lrgarol 1051, Diuron and Sea-Nine 211, respectively. The high mortality rate was Sea-Nine 211, Diuron and lrgarol 1051, respectively. A. omorii has a higher hatching rate and lower mortality rate than P. parvus s.1., indicating that A. omorii has a higher tolerance for the toxicity of biofouling debris from ships’ surface. The LC₅₀ of P. parvus s.l. in 24h-toxicity test was 2.17 ppm for Diuron, 1.24 ppm for lrgarol 1051, and 0.007 ppm for Sea-Nine 211, indicating that Sea-Nine 211 has considerably higher toxicity than other active substances. The LC₅₀ values of P. parvus s.1. for CuPT and ZnPT are 6.16 ppb and 1.19 ppb, respectively, which is lower than the LC₅₀ of CuPT and ZnPT. It means that CuPT and ZnPT is more toxic than CuPT and ZnPT.

c. Identification and investigation of risk factors for biofouling debris from ship’s surface during in-water cleaning activities
(1) Collection, identification and biological risk assessment of ship’s biofouling
(a) Fouling of diatoms on the bottom of ship’s surface
Abundances of fouling diatoms on the ships’ surfaces: Local(R/V JANGMOK 1 and 2) 〉 Coastal(R/V EARDO) > Ocean(R/V ONNURI). Pre-dominant diatom was Navicula muscatinei.

(b) Occurrence of protozoa
A total of six ciliate species was detected from the four vessels； one species from JANGMOK 1 (Loxophyllum rostratuni), two species from JANGMOK 2 (Euplotes parked E. plicatuni), one species from EARDO (Amphisiella annulata), and two species from ONNURI (Uronychia binucleata, Arcuseries petzi). There was no ciliate occurrence from ISABU surface. Euplotes parkei and E. plicatum isolated from JANGMOK 2 are unknown species in Korea. This result means that these species may be invasive ciliates originated from ship transportation. Further analyses should be added to confirm the invasive reality.

(c) Fouling of macrozoobenthos on the bottom of ship’s surface
A total of 47 species was collected with a mean density of 10,297 indi./m² and a mean biomass of 2,381 gWWt/m², respectively. Crustaceans were the dominant taxon in terms of species richness (24 species, 51.1%), and followed by polychaetes (9 species, 19.1%) and molluscs (6 species, 13.2%). The mean density were highest in the crustaceans in which the Balarms species appeared in the most abundant species, but the highest in biomass in the molluscs. The number of species of macrobenthos on R/V EARDO was highest in 25 species, and 18 species and 19 species were found in R/V JANGMOK 2 and R/V JANGMOK 1, respectively. However, the number of species at R/V ONNURI and R/V ISABU decreased to 9 species and 8 species, respectively. Fouling of macrozoobenthos at R/V JANGMOK 1 and R/V EARDO appeared in higher density and biomass. Macrozoobenthos at R/V ONNURI showed the highest density and biomass in the flat bottom of the hull, which Balanus improvisus and B. amphitrite were the dominant species. Macrozoobenthos at R/V EARDO found in the most abundant species (17 species) on propeller shafts of niche areas and showed high biomass. The density at R/V EARDO was higher in the stem thruster. The dominant species were B. amphitrite, dominant on the hull side and bottom, and B. trig-onus, dominant on the left thruster of the athlete of R/V EARDO. In R/V JANGMOK 2, 14 species were found in the upper part of the left side, and the high density was on the bottom area. Biomass was higher in the upper side of the ship and in the left propeller shaft. The dominant species were B. trigonus and H. ezoensis. In case of R/V JANGMOK 1, 9-12 species appeared on the side and bottom of the hull, showing high density. Biomass was higher in the propeller shaft. B. amphitrite showed dominant dominance and relatively high density distribution in the middle area of the side. R/V ISABU was found to have 2-4 species on the side and bottom of the hull, showing low density. Density and biomass was higher at the propeller shaft. The dominant species were B. amphitrite and B. trigonus.

(2) Identification of biological debris from simulated experiment
(a) Diatoms from simulated experiment
Abundances of attached fouling diatoms: control plate (Max. 283,533 cells cm^-2) > treatment plate (Max. 45,130 cells cm^-2). Fouling diatoms from simulated experiment (viability test): continuous increase in cell density of fouling diatoms (biological debirs) in control and treatment groups.

(b) Species change of protozoa
Four ciliate species (Aspidisca leptaspis, Diophrys appendiculata, Euplotes parked and Protogastrostyla pulchra) were observed on the simulated plates. Species composition of ciliate fauna showed different patterns until 4 weeks between treatments and background plates. After 5 weeks, the fauna changed gradually to similar composition on both simulated plates. D. appendiculata and E. parkei appeared dominantly in the simulated plate respectively. These two species is sustained in culture strains.

(c) Macrozoobenthos from simulated experiment
Dominant macrozoobenthos on the background plate of simulated experiment were mainly Ciona intestinalis, Ascidiella aspersa, Bugula neritina and Balanus trig onus, which were mainly attached to the treatment plate, analyzing of biofouling index of attached macrobenthos, we found that stage III appeared in the background plate and stage I in the treatment plate after 1 month. After two months, stage V was in the background simulator and stage I in the treatment plate. After 3 months, stage V was showed in the background plate and 5 stage in the treatment plate. After 4 months, the background plate had 4 stage and the treatment plate had stage IV. In Secondary biofilm-1 experiments, many species, Ciona intestinalis, Bugula neritina” Sponges, attracted on two background plates (B1 and B2), but only the biofilm found on the two treatment plates (T1 and T2). Ascidiacea of B1 and B2 died more than 90% on the 2^nd day after the experiment and survived until the 5^th day. Sponges and Bryozoa died 100% on the 2^nd and 3^rd day, respectively. In case of Secondary biofilm-2 experiments, macrozoobenthos on two backbround plates (B1 and B2) appeared not only biofouling species such as Ciona intestinalis, Ascidiella aspersa, Bugula neritina, sponges but also mobile species such as amphipods, isopods, polychaetes. Only the biofilm was attached at the treatment plates (T1 and T2). All of mobile species died on the second day after the experiment, but Ascidiacea survived until the 5^th day after the experiment. All sponges died on the 2^nd day and Bryozoa survived 50% until the 41^st day of the experiment. In secondary biofilm-3 experiments, the background plates (B1 and B2) were attached not only biofouling species, Ciona intestinalis, Ascidiella aspersa, sponges, Actiniaria, and Bryozoa but also mobile species, amphipods, Isopods, and polychaetes. However, the 75% of density of barnacles appeared at the treatment plates (T1 and T2). All mobile species died on the 4^th day after the experiment, but 30% of attached Ascidiacea contiuned to survive until 30 days. All sponges and Bryozoa died on the 4^th day and the 9^th day after the experiment, respectively. In treatment plates (T1 and T2), 50% of the damaged barnacles died on the first day, but small amount of species (2.5%) survived until the end of the experiment. In secondary biofilm-4 experiments, the background plates (B1 and B2) were attached not only biofouling species, Ciona intestinalis, Ascidiella aspersa, sponges, Actiniaria, Bryozoa, but also mobile species, amphipods, isopods and polychaetes. Actiniaria and barnacles appeared to more than 65% of total density on the treatment plates (T1 and T2). In this experiment, we added the washed sand into the two experiment baths (T1, B1) and sterilized mud into the other baths (B2 and T2). All the mobile species of B1 and B2 died on the 5^th day of the experiment, but Ascidiacea survied until the 33^rd day. There was no significant difference in the survival rate between two sediment conditions. The barnacles on the treatment plates (T1 and T2) died on the 34^th day after the experiment, and the individuals buried in the sediments died within a short time after begining the experiment. However, in all water tanks, breeding of Actiniaria showed active movement and survived until the end of the experiment.

d. Selection of the key management factors and suggestion of framework for the in-water cleaning management
(1) Exploration of scenarios for in-water cleaning activities
Australia and New Zealand carry out the most systematically and strictly performing the management of underwater hull cleaning. They enacted the Craft Risk Management Standard (CRMS) in 2014, and is being piloted for four years. It will be enforced in May 2018. Most other European countries did not have no specific regulation of the management of underwater hull cleaning. Some European countries carry out underwater hull cleaning using a dry-dock in accordance with IMO’s regulation for removing fouling organism. Removal techniques of biofouling organism are classified into manual removal, mechanical removal, surface treatment, and shrouding technique. In case of large commercial vessels moving between countries, mechanical removal and surface treatment methods are mainly used to remove biofouling organism. However, the majority of mechanical removal method, including electric brushes, can be to affect the marine environment by fragments or leached active substances of AFS, and furthermore there is no collection device for removing AFS derivatives.

(2) Derivation of key elements for the management level
The most important factors are systematically manage to the underwater hull cleaning and the present condition of arrival and departure in domestic port together in the comprehensive review of the documents reported by Australia and New Zealand related to remove biofouling organism. The selected main keys determining the management of underwater hull cleaning are biofouling origin, anti-fouling coating (AFC) type, biofouling type, cleaning method, and debris capture. We have created 40 scenarios to remove biofouling organism based on selected risk assessment/management key factors. Based on the prepared scenarios to remove biofouling organism, we plan to carry out biological and chemical risk assessment using assessment indicators.

(3) Suggestion of risk assessment technique for the in-water cleaning management
The International Maritime Organization (IMO) has recognized the risk of hull fouling and announced ‘2011 Guidelines for the control and management of ship’s biofouling to minimize the transfer of invasive aquatic species’ and is planning international regulations to enforce them in the future. In this study, to effectively respond to future international regulation, we introduce the case of leading countries related to management of hull fouling and also investigate environmental risk assessment techniques for in-water cleaning. Australia and New Zealand, the leading countries in hull fouling management, have established hull fouling regulations through biological and chemical risk assessment based on in-water cleaning scenarios. Most European countries without their government regulation have been found to perform in-water cleaning in accordance with the IMO’s hull fouling regulations. In the Republic of Korea, there is no domestic law for hull fouling organisms, and only approximately 17 species of marine ecological disturbance organisms, are designated and managed under the Marine Ecosystem Law. Since in-water cleaning is accompanied by diffusion of alien species and release of chemical substances into aquatic environments, results from biological as well as chemical risk assessment are performed separately, and then evaluation of in-water cleaning permission is judged by combining these two results. Biological risk assessment created 40 codes of in-water cleaning scenarios, and calculated Risk Priority Number (RPN) scores based on key factors that affect intrusion of alien species during in-water cleaning. Chemical risk assessment was performed using the MAMPEC (Marine Antifoulant Model to Predict Environmental Concentrations), to determine PEC and PNEC values based on copper concentration released during in-water cleaning. Finally, if the PEC/PNEC ratio is >1, it means that chemical risk is high. Based on the assumption that the R/V EARDO ship performs in-water cleaning at Busan’s Gamcheon Port, biological risk was estimated to be low due to the RPN value was <10,000, but the PEC/PNEC ratio was higher than 1, it was evaluated as impossible for in-water cleaning. Therefore, it will be necessary for the Republic of Korea to develop the in- water cleaning technology by referring to the case of leading countries and to establish suitable domestic law of ship’s hull fouling management in domestic harbors.

(출처 : SUMMARY 31p)

목차 Contents

• 표지 ... 1
• 제 출 문 ... 2
• 보고서 초록 ... 3
• 요 약 문 ... 7
• SUMMARY ... 26
• CONTENTS ... 49
• 목차 ... 50
• 그림목차 ... 51
• 표목차 ... 61
• 제 1 장 서 론 ... 64
• 제 1 절 연구개발의 필요성 ... 64
• 제 2 절 연구개발의 목표 및 내용 ... 70
• 제 2 장 국내외 기술개발 현황 ... 72
• 제 1 절 국내 연구동향 ... 72
• 제 2 절 국외 연구동향 ... 73
• 제 3 장 연구개발수행 내용 및 결과 ... 75
• 제1절 AFS 처리활성물질 분석기술개발 ... 75
• 제2절 선박부착 파생물의 LAB-SCALE영향평가, 지표발굴, 모사 실험 및 AFS활성물질 영향 평가 ... 109
• 제3절 선박부착기인 생물 및 위해성 요인탐색 및 판별시도 ... 192
• 제4절 수중제거 핵심 관리요소 선정 및 관리 framework제시 ... 278
• 제 4 장 연구개발목표 달성도 및 대외기여도 ... 300
• 제 1 절 목표 달성도 ... 300
• 제 2 절 대외 기여도 ... 304
• 제 5 장 연구개발결과의 활용계획 ... 306
• 제 6 장 참고문헌 ... 308
• Appendix ... 316
• 끝페이지 ... 328