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Effect of light emitting diode (LED) on viral hemorrhagic septicemia (VHS) and growth in olive flounder

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Alternative Title
LED (발광 다이오드)의 넙치에 대한 Viral hemorrhagic septicemia (VHS)와 성장 효과
Abstract
Although previous studies have intensively studied about antimicrobial effects of blue light on bacteria and parasite, little is known about the antimicrobial effects of blue light on virus. In addition, green LED light has been proved to have great growth promotion effects on some fish species but there is limited information about the effects of green LED light on growth performance of olive flounder. This study was aimed to determine the effects of blue LED light on viral hemorrhagic septicemia virus (VHSV) and on VHS in olive flounder (Chapter 1), growth promotion effects of green LED on olive flounder (Chapter 2), and their adverse effects (Chapter 3). In chapter 1, VHSV (in vitro) and olive flounder infected with VHSV (in vivo) were exposed to 405 nm LED with the light intensity of 10, 20, 30, and 40 µ·mol∙m-2∙s-1 at 15°C. Results show that blue LED light successfully inactivated VHSV with a dose-dependent manner. In in vivo experiment, virus concentration in rearing water and survival rate of fish exposed to 405 nm LED groups whose light intensity were larger than 20 µ·mol∙m-2∙s-1 were higher than those in the control groups, suggesting that 405 nm with the light intensity of 20 µ·mol∙m-2∙s-1 or more could be used to inactivate virus in rearing water, prevent disease transmission and increase fish survival rate. In chapter 2, olive flounder were exposed to ambient light, 405 nm, 520 nm, and mixed lights (450 nm and 520 nm). There were two experiments including small-scale (8 weeks) and pilot-scale (12 weeks). Results show that growth performance of olive flounder exposed to 520 nm LED and mixed lights in either 8 weeks or 12 weeks were significantly higher than that of fish under 405 nm LED and ambient light. The expression level of growth hormone in fish under green light was significantly higher compared to that of fish in other groups. Chapter 3 focused on determining the adverse effects of LEDs on olive flounder with the same wavelengths and experimental design as chapter 2. Results show that there were no significant differences in the expression levels of stress and immune-related genes, serum glucose level and eye histopathological morphology when olive flounder exposed to LEDs in small-scale and pilot-scale. In conclusion, this study presents antimicrobial effects of blue LED on virus, growth promotion effects of green LED on olive flounder with limiting adverse effects, suggesting possible eco-friendly treatment method to future fish farming system.
이전에 청색광 LED에 대한 항미생물효과와 항기생충효과는 많이 연구되었지만, 항바이러스효과에 대한 연구는 아직까지 미비하다. 또한, 녹색광 LED에서 여러 종류의 어류의 성장을 촉진시킨다고 알려져 있지만, 아직까지 국내에서 많이 양식되는 넙치에 대한 연구결과는 부족한 실정이다. 본 연구의 목적은 청색광 LED의 조사가 바이러스 출혈성 패혈증 (Viral hemorrhagic septicemia) 의 원인체인 VHSV에 미치는 영향에 대해 확인하였고 (Chapter 1), 녹색 LED의 조사가 넙치의 성장에 미치는 영향에 대해 조사하였으며 (Chapter 2), 사용된 LED 광조건에 대한 유해성 평가를 실시하였다 (Chapter 3). Chapter 1에서는 VHSV를 15 10, 20, 30 그리고 40 µ·mol∙m-2∙s-1 의 광량으로 청색광의 LED를 15°의 배양조건에서 조사해주었을 때, VHSV가 농도의존적으로 사멸되는 것을 확인하였다 (In vitro). 해당 조건을 적용하여 생체실험 (In vivo)을 진행했을 때는 20 µ·mol∙m-2∙s-1 이상의 광량에서 상대적으로 낮은 농도의 VHSV가 사육수에서 검출되었으며 생존률이 높아지는 것을 확인하였다. 이는 405nm를 20 µ·mol∙m-2∙s-1 이상으로 조사 할 때 바이러스의 전파를 예방하고 어류의 생존률이 개선 될 수 있다는 것을 의미한다. Chapter 2에서는 자연광, 405nm, 520nm 그리고 복합광 (405+520nm)을 넙치에 조사해 주었다. 실험실 규모의 실험을 8주간 진행하였으며, pilot 규모의 실험은 12주간 수행하였다. 그 결과 넙치의 성장이 8주 또는 12주간 520nm와 복합광을 조사한 구간에서 405nm와 자연광을 조사한 구간 대비 유의적으로 증가하는 것을 확인하였으며, 성장호르몬의 유전자 발현도 520nm에서 유의적으로 증가하는 것을 확인하였다. Chapter 3에서는 LED 조사시 발생할 수 있는 잠재적인 유해성의 유무에 대해 판단하였다. 유해성 평가에 사용된 실험 조건은 Chapter 2. 에서 사용한 조건과 동일하게 설정하였다. 그 결과, 어떤 구간에서도 스트레스, 면역 관련 유전자, 혈청 내 glucose 농도, 안구의 형태학적인 변화의 유의적인 발현 차이가 실험실 규모의 실험과 pilot 규모의 실험 모두에서 확인되지 않았다. 결론적으로, 본 연구에서는 청색광의 항바이러스 효과와 녹색광의 성장 효과를 확인하였으며, 여기에 대한 유해성이 거의 존재하지 않는다는 것을 확인하였으며, 본 연구의 결과는 친환경적 질병 치료 방법 및 미래 양식 시스템의 적용에 크게 기여할 것으로 판단된다.
Author(s)
HO DIEM THO
Issued Date
2019
Awarded Date
2019. 2
Type
Dissertation
Keyword
LED VHSV Olive flounder Growth performance
Publisher
부경대학교
URI
https://repository.pknu.ac.kr:8443/handle/2021.oak/23105
http://pknu.dcollection.net/common/orgView/200000182606
Affiliation
부경대학교 대학원
Department
대학원 수산생명의학과
Advisor
김도형
Table Of Contents
Chapter 1 1
Effect of blue light emitting diode (LED) on viral hemorrhagic septicemia (VHSV) and on VHS in olive flounder 1
1. Introduction 2
1.1. Viral haemorrhagic septicaemia (VHS) 2
1.2. Current strategy of light therapy for controlling virus 2
1.3. Light emitting diodes 4
2. Methodology 5
2.1. Light sources 5
2.2. Virus amplification and quantification 6
2.2.1. Virus strain used in this study 6
2.2.2. Virus amplification 6
2.2.3. Plaque assay 7
2.3. Anti-viral activity of 405 nm with different light intensities 8
2.4. Effect of 405 nm with different light intensities on olive flounder infected by VHSV 8
2.5. Statistical analysis 10
3. Results 11
3.1. Anti-viral activity of 405 nm with different light intensities 11
3.2. Effect of 405 nm with different light intensities on olive flounder infected with VHSV 13
3.2.1. Survival rate 13
3.2.2. Viral concentration in rearing water 19
4. Discussion 22
5. Conclusion 25
Chapter 2 27
Effect of green light emitting diode (LED) on somatic growth in olive flounder 27
1. Introduction 28
2. Methodology 29
2.1. Light sources 29
2.2. RNA extraction, cDNA synthesize and real-time PCR 29
2.3. Somatic growth of olive flounder under different wavelengths of LEDs 31
2.4. Statistical analysis 33
3. Results 35
3.1. Growth performance 35
3.2. Expression of growth-related genes 39
4. Discussion 41
5. Conclusion 44
Chapter 3 45
Harmful effects of LED light with different wavelengths on olive flounder 45
1. Introduction 46
2. Methodology 47
2.1. Light sources 47
2.2. RNA extraction, cDNA synthesize and real-time PCR 47
2.3. Side effects of LED with different wavelengths on olive flounder 50
2.4. Statistical analysis 52
3. Results 54
3.1. The expression level of stress and immune-related genes 54
3.2. Serum glucose level 54
3.3. Eye histopathology 54
4. Discussion 59
5. Conclusion 60
국문요약 63
Acknowledgment 65
References 67
Degree
Master
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대학원 > 수산생명의학과
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