초록 ▼

○ 물렁증 발생 하한수온은 6.9℃이상, 용존산소 반수치사농도 3.55ppm, 반수치사 염분농도 25.4psu으로 분석되었으며, 42일간 절식시험결과 물렁증 발생률은 15%으로 단기간내 저산소, 저염분 및 절식 등 환경요인에 의한 물렁증 발생은 희박한 것으로 추정됨

○ 멍게의 대량폐사원인 구명 및 폐사방지 방안 마련을 위해 양식가이드북「멍게양식과 물렁증 예방대책」을 발간하여 어업인들에게 보급

○ 멍게 물렁증원인체로서 동원핵편모충 Azumiobodo hoyamushi, 미생물 2종 Hasllibacter ha

○ 물렁증 발생 하한수온은 6.9℃이상, 용존산소 반수치사농도 3.55ppm, 반수치사 염분농도 25.4psu으로 분석되었으며, 42일간 절식시험결과 물렁증 발생률은 15%으로 단기간내 저산소, 저염분 및 절식 등 환경요인에 의한 물렁증 발생은 희박한 것으로 추정됨

○ 멍게의 대량폐사원인 구명 및 폐사방지 방안 마련을 위해 양식가이드북「멍게양식과 물렁증 예방대책」을 발간하여 어업인들에게 보급

○ 멍게 물렁증원인체로서 동원핵편모충 Azumiobodo hoyamushi, 미생물 2종 Hasllibacter halocynthiae, Pseudoalteromonas elyakovii을 구명하였음

○ 물렁증원인체인 미생물구제제로서 굴에서 추출한 항균물질 Homarine 개발

○ A. hoyamushi의 최적 성장 범위는 5-10℃, 30-35 psu으로 나타났음

○ 물렁증 원인체로서 바이러스 검출안됨

○ 현재까지 존재하는 존재하는 모든 멍게관련 정보와 현 연구를 통해 생산되어진 멍게 유전자 정보를 바탕으로 멍게 유전자 정보 DB 구축

○ 기생충구제제를 위한 약욕시험결과 F:H 8:2 ㎍/㎖의 농도로 1시간 약욕했을 때 멍게에 대한 독성이 낮고 기생충 구제 효과가 높았음

○ 동해안 지역의 물렁증 발생은 주로 5∼6월에 발생하며, 물렁증원인체는 남해안 양식멍게와 동일한 것으로 추정됨

(출처 : 보고서 요약 2p)

Abstract ▼

Ⅵ. Results
1. Tolerance range by environmental factors and biomarker development
○ Water temperature (35days-LT50) which causes more than 50% of softness syndrome was 6.9℃ 3.4∼22.5℃), whereas fatal high temperature 30days-LT50) was identified as 21.2℃ 16.7∼23.6℃).
○ 5days-LT50 was 3.55 (.86

Ⅵ. Results
1. Tolerance range by environmental factors and biomarker development
○ Water temperature (35days-LT50) which causes more than 50% of softness syndrome was 6.9℃ 3.4∼22.5℃), whereas fatal high temperature 30days-LT50) was identified as 21.2℃ 16.7∼23.6℃).
○ 5days-LT50 was 3.55 (.86-4.96ppm) and soft-tunic syndrome was not found in short-term acute hypoxia.
○ 8 days-LS50 was 25.4psu and after 42 days fasting, 15% of soft-tunic syndrome and 50% of survival rate was shown respectively, thus environmental factors in the short-term have little affect on the syndrome.

2. Study on healthy seed production and improvement of nurturing method
○ We published a Sea Squirts Aquaculture and Preventative Measures of Soft-tunic Syndrome and distributed it to fishermen to find out causes of mass mortality of the species and establish preventative measures.
○ 500 fascines of healthy seed were collected and transplanted in Tongyoung, Gyeongsangnam-do, in March and they are under cultivation. Test results in Nov. showed 1.62 individuals attachment, 10.2mm of body length, 0.41g of total weight.
○ To improve nurturing method, we are operating a test fishing ground (20ha of size, 40-50m of water depth) in Nodae waters, Yokji-eup, Tongyoung.

3. Study on microparasite of sea squirts and their influence on death
○ In the aerotropic condition, we separated 250 strains of microorganisms from normal sea squirts and 293 strains from abnormal sea squirts. In the anaerobic condition, we separated 25 strains of microorganisms from normal sea squirts and 26 strains from abnormal sea squirts. 24 strains from the aerotropic condition and 6 strains from the anaerobic condition were separated from the collecting area.
○ There's no significant difference between bacterial groups of normal and abnormal (soft-tunic) sea squirts. Also, seasonal effect of bacterial groups did not appear.
○ After repetitive studies on the death effect of dominant microorganisms separated from soft-tunic sea squirts, Strain KME 002T, AD 37 group showed death after 13 days which is similar to the death after softness syndrome and was evaluated as a major cause of the syndrome. After identification, KME 002T was named as Hasllibacter halocynthiae, and Strain AD 37 as Pseudoalteromonas elyakovii.
○ To reduce the death rate of sea squirts, we developed Homarine which is extracted from oyster and has an antibacterial effect on soft-tunic microorganisms.

4. Establishment of ormics research base for sea squirtss with soft-tunic syndrome
○ Analysis study between normal and abnormal sea squirtss after establishing the cDNA library and searching with transcriptomical method showed an increase in calponin, muscle actin in the abnormal group and decrease in halocidin.
○ We found out four suspicious infection agents after establishing a genomic library, analyzing end-sequencing and bio-information.
○ Based on end-sequencing of abnormal sea squirt's genomic library, we revealed

4 suspicious clones.
○ After reading infected dielectrics and collecting all the ESTs, PCR product in K07 was detected.
○ In sea squirts with the syndrome, cause transfectant was extracted via metagenomic analysis and explored a candidate idioblast primer for infection diagnosis.
○ Based on all research on sea squirts so far plus genetic information of the species through this study, sea squirt genetics information DB has been made.
○ Development of soft-tunic syndrome diagnosis kit-By developing diagnosis kit with LAMP, faster diagnosis on flagellate with small amount than PCR can be achieved .

5. Investigation of the causes of soft-tunic syndrome and death mechanism
○ Protozoan which has 2 flagellum in the film was found, and 97.5% of infection rate was shown, detected specifically in soft-tunic sea squirts.
○ Flagellate was separated and then injected to healthy sea squirts. As a result, specific soft-tunic syndrome occurred.
○ According to phylogenetic systematic research, the causing agent of the syndrome was Azumiobodo hoyamushi, a kind of kinetonucleus flagellate.
○ A. hoyamushi showed optimum growth at 5-10℃, 30-35psu but it slowed down in growth or died if it was not within the range.
○ Based on real-time PCR measurement of A. hoyamushi by film part, we found out parasite level at exhalent/inhalent opening was notably higher than others, which means they play an important role in early infection.
○ It was reported that Formalin (F) is useful for killing A. hoyamushi, especially the combination of F and hydrogen peroxide (H) functions 3 times than the sole chemicals.
○ Vivo test targeting soft-tunic sea squirts revealed that F:H 8:2 ㎍/㎖ ratio for 1 hour was the most effective for disinfestation and has the lowest toxicity.

6. Study on soft-tunic syndrome from cultured sea squirts in the East Sea
○ Softness syndrome in the East Sea usually happens in May to June when cold water occurs frequently, with big difference in water temperature between surface and bottom.
○ 30day-LT50 of sea squirts in the East Sea was 22.16℃ (19.08∼26.16)℃ and 7day-LS50 was 24.79psu (24.08∼5.37psu).
○ The agent causing the softness syndrome was considered to be the same in the East and Southern Sea.

(출처 : SUMMARY 9p)

목차 Contents

• 표지 ... 1
• 보고서 요약 ... 2
• 요약문 ... 3
• SUMMARY ... 9
• 목차 ... 16
• CONTENTS ... 19
• 그림목차 ... 22
• FIGURE LISTS ... 26
• 표목차 ... 30
• TABLE LISTS ... 32
• 제 1 장 연구개발과제의 개요 ... 34
• 제 2 장 국내외 기술개발 현황 ... 35
• 제 3 장 연구개발 수행내용 및 결과 ... 36
• 제 1절 연구개발 수행 방법 ... 36
• 1. 환경요인별 내성범위 및 바이오마커 개발 ... 36
• 2. 건강종묘생산 및 양성방법 개선 ... 36
• 3. 멍게 기생 미생물 분석 및 폐사영향성 구명 ... 37
• 4. 물렁증에 걸린 멍게의 오믹스연구 기반 구축 ... 38
• 5. 멍게물렁증 원인체 탐색 및 폐사 기작 연구 ... 41
• 6. 동해안 멍게 물렁증 원인 탐색 ... 51
• 7. 물렁증 진단키트 개발 및 제작 고리매개 등온증폭반응 loop-mediated isothermalamplication, LAMP 을 이용한 검출법 ... 51
• 제 2절 연구개발 수행 결과 및 고찰 ... 52
• 1. 환경요인별 내성범위(수온, 절식, 염분, 용존산소) 및 바이오마커 개발 ... 52
• 2. 건강종묘생산 및 양성방법 개선 연구 ... 63
• 3. 멍게 기생미생물 분석 및 폐사영향성 연구 ... 64
• 4. 오믹스기반 구축을 통한 멍게 물렁증 감염체 탐색 ... 76
• 5. 멍게 물렁증 원인체 탐색 및 폐사 기작 연구 ... 87
• 6. 동해안 멍게 물렁증 원인 탐색 ... 104
• 7. 물렁증 진단 키트 개발 및 제작 ... 106
• 제 4 장 목표달성도 및 관련분야에의 기여도 ... 115
• 제 1절 목표달성도 ... 115
• 1. 멍게 환경내성범위 및 바이오마커 개발 및 양식방법 개선 ... 115
• 2. 멍게 기생미생물 분석 및 페사영향성 연구 ... 115
• 3. 물렁증에 걸린 멍게의 오믹스 연구기반 구축 ... 115
• 3. 멍게 물렁증 원인체 탐색 및 폐사 기작 연구 ... 115
• 4. 동해안 양식멍게 물렁증 탐색 연구 ... 116
• 제 2절 관련분야에의 기여도 ... 116
• 제 5 장 연구개발결과의 활용계획 ... 116
• 제 6 장 참고문헌 ... 117
• 끝페이지 ... 121