표제지
목차
요약문 4
SUMMARY 5
제1장 연구개발과제의 개요 9
제1절 연구개발의 목적 9
제2장 국내외 기술개발 현황 10
1. 국내 기술개발 현황 10
2. 국외 기술개발 현황 10
제3장 연구개발 수행내용 및 결과 11
제1절 어미 관리 기술 개발 11
1. 어미 후보군 확보 및 관리 기술개발 11
2. 기초 번식내분비 특성 조사 35
3. 조기 성숙유도 기술 개발 41
4. 수정란 생산 및 생산조건 탐색 44
5. 실내 월동 양성 및 성숙 유도 시험 49
제2절 종자생산 기술 개발 50
1. 부화자어 적정 사육환경 요인 탐색 50
2. 난질평가 인자 탐색 54
3. 종자 발달단계별 성장 및 질병 모니터링 59
4. 종자 생존율 향상 연구 65
제3절 중간육성 고도화 기술 개발 68
1. 실내 중간육성 조건 탐색 68
2. 양성방법별 양성효과 분석 70
제4장 목표달성도 및 관련분야에의 기여도 72
제5장 연구개발결과의 활용계획 73
제6장 참고문헌 74
제7장 부록 75
판권기 76
Table 1. Transportation of wild pomfret broodstock by different collection methods (2020) 14
Table 2. Results of 6 hours-transportation of wild broodstock from adjusted in sea cage (7th, Aug. 2020.) 14
Table 3. Results of 6 hours-transportation of wild broodstock under different stock density (30th~31st, Oct. 2020) 15
Table 4. Collection of juveniles and broodstock of pomfrets and their tansportation(6 hours, 2021) 16
Table 5. Rearing status for juveniles and broodstock of pomfrets (31st, Dec. 2021) 16
Table 6. Survival rate for transportation according to stock density 19
Table 7. Sex ratio of pomfrets broodstock 20
Table 8. broodstock management of Korean pomfret (at the end of Dec., 2022) 20
Table 9. Comparison of survival rate for broodstock followed anesthetic concentration, low water temperature treatment and different stock densities during transportation 21
Table 10. Investigation of feed preference for pomfret broodstock 23
Table 11. Investigation for feed reaction of broodstock and juvenile pomfrets 25
Table 12. Disease monitoring for dead broodstock during transporation and adjustment in indoor tank 26
Table 13. Disease monitoring of dead fish during transportation and rearing in indoor tank 27
Table 14. Bacterial monitoring of seawater and rearing water 27
Table 15. Monthly number of bacterial cells from rearing water of broodstock 28
Table 16. Analysis of relationship among morphometrics in pomfrets 34
Table 17. Species-specific genetic marker for pomfrets 34
Table 18. Morphometric and genetic analysis for pomfrets from each site 35
Table 19. Analysis of species-specific genetic marker for identification 35
Table 20. Monthly sex ratio of indoor-reared pomfrets 37
Table 21. Fecundity of pomfrets according to collection period and morphometric characteristics 39
Table 22. Maturity of broodstock during wintering 42
Table 23. Gonadal development of broodstock during rearing period (Dec., 2021 to July, 2022) 44
Table 24. Summary of natural spawning from indoor reared broodstock 46
Table 25. Comparison of artificial egg and sperm collection according to maturation stage 48
Table 26. Summary of artificial spawning from wild broodstock 48
Table 27. Comparison of fertilization rate according to different methods 51
Table 28. Fertilization and hatching in artificial egg-collection 52
Table 29. Results for artificial collection of eggs and hatching in P. argenteus 52
Table 30. Monthly changes in total length, body weight and GSI of wild P. argenteus 53
Table 31. Comparison of total amino acid contents in different organs of pomfrets 55
Table 32. Comparison of fatty acid in different organs of pomfrets 56
Table 33. Composition of fatty acid composition in different organs of pomfrets 57
Table 34. Composition of fatty acid between unfertilized and fertilized eggs of P. argenteus 58
Table 35. Summary of seed production for P. argenteus during 2020~2022 63
Table 36. Weekly changes in rearing water condition for hatched larvae P. argenteus 64
Table 37. Comparison of survival rate and total length in hatched larvae P. argenteus among different fee supply treatment 65
Table 38. Effects of food supply on growth and survival of P. argenteus larvae 69
Table 39. Effects of high water temperature treatment on survival of hatched larvae 69
Table 40. Comparison of rearing water condition between flow-through and RAS intermediate culture 71
Fig. 1. Establishment of double fish tank for broodstock management. (A) scheme of the tank, (B) setting of tank 11
Fig. 2. Collection methods and adjustment place for wild pomfret broodstock. (A) fence net, (B) stow net, (C) set net, (D) sea cage 12
Fig. 3. Establishment of double circular fish tank for broodstock management. (A) rearing system, (B) scheme of the rearing system (① boiler, ② heat exchanger,... 12
Fig. 4. Rearing condition for adjustment in indoor tank of pomfret broodstock. (A) survival rate, (B) water temperature, (C) salinity, (D) DO, (E) pH 13
Fig. 5. Transportaion process of wild pomfret broodstock by fence net. (A) fence net, (B) collection by fence net, (C) transportation, (D) non-exposure transportation 14
Fig. 6. Changes in water quality during transportation of pomfret broodstock. (A) ammonia-N, (B) nitrite-N, (C) nitrate-N, (D) total nitrogen 15
Fig. 7. Collection process of pomfrets broodstock and adjustment of indoor tank. (A) set net collection, (B) macroscopy of ceollected fish, (C) Survival inspection... 16
Fig. 8. Changes in survival and status of rearing broodstock of pomfrets. (A) rearing of juveniles, (B) rearing of broodstock, (C) successfully adjusted fish in indoor tank,... 17
Fig. 9. Changes in rearing water quality during adjustment in indoor tank. (A) water temperature, (B) DO, (C) salinity 17
Fig. 10. Changes in water quality factors from rearing water during adjustment in indoor tank 18
Fig. 11. Changes in water quality during transportation according to stock density. (A) ammonium, (B) nitrite, (C) nitrate, (D) TN; E, DIN, (F) TP, (G) phosphate,... 18
Fig. 12. Relationship among factors for transportation of pomfrets 20
Fig. 13. Comparison of (A) changes in water temperature and (B) dissolved oxygen levels during transportation time for each treatment 21
Fig. 14. Comparison of water quality factors from water followed anesthetic concentration, low water temperature treatment and different stock densities after... 22
Fig. 15. Manufacturing process of gelatin feed for pomfret 23
Fig. 16. Stomach contents of pomfret and supplied shrimp. (A) stomach contents (shrimp), (B) stomach contents (shrimp, before digestion), (C) supplied mysids... 24
Fig. 17. Applied feeds for adjustment of pomfret broodstock. (A) submerging feed for flounders, (B) paste feeds for eels, (C) frozen mysids 24
Fig. 18. Comparison of characteristics for agar, bacto-agar, gelatin and supply of gelatin feed to broodstock. (A) agar, (B) bacto-agar, (C) gellatin, (D) gellatin-coated feed 25
Fig. 19. External features of dead fish and its disease monitoring. (A) mortality after transportation (scale peeling), (B) disease inspection of internal organs from... 26
Fig. 20. Symptoms for disease of dead pomfrets reared in indoor tank with flow-through system. (A) peeled epidermis, (B) hemophthalmus, (C). observation of Anisakis spp 27
Fig. 21. Changes in water temperature, salinity and survival of broodstock. (A) water temperature, (B) salinity and survival rate 29
Fig. 22. Survival rate and water temperature at each stock density. (A) survival rate, (B) water temperature at each density treatment 29
Fig. 23. Changes in water quality at each stock density for pomfret broodstock. (A) ammonium, (B) nitrite, (C) nitrate, (D) total nitrogen 30
Fig. 24. Comparison of (A) dissolved oxygen levels (B) survival rate and (C) number of dead individuals after transportation of broodstock with or without oxygen supply 31
Fig. 25. Process of species identification for Pomfrets by fish size. (A) image shooting, (B) calibraion for factor analysis, (C) coding and statistical analysis, (D) factor analysis 31
Fig. 26. Changes in morphology with growth of P. argenteus. (A) straight to curved as the upper curved surface of the head grows, (B) as the fin rays grow they gradually... 32
Fig. 27. Image analysis of morphometrics of pomfrets 32
Fig. 28. Results of factor analysis for morphometric images of pomfrets. (A) statistical analysis, (B) reversed-image matrix, (C) Total variation, (D) scree table,... 33
Fig. 29. Changes in GSI and HSI of different size of wild pomfrets. (A) GSI for middle size group, (B) HSI for middle size group, (C) GSI for large size group,... 36
Fig. 30. Histological observation of ovarian (left) and testicular (right) development. (A) early growing, (B) late growing, (C) maturation, (D) ripe 37
Fig. 31. Quantitative analysis of E2 and T levels in ovary of pomfrets collected in 2020. (A) E2, (B) T 38
Fig. 32. Quantitative analysis of 11-KT and T levels in testis of pomfrets collected in 2020. (A) 11-KT, (B) T 38
Fig. 33. Plasma or gonadal levels of sex steroids of Pomfret during reproductive period from 2021. (A) plasma levels of estrogen (E2), (B) plasma levels of Testosterone,... 39
Fig. 34. Process of species identification for pomfrets. (A) image shooting, (B) calibraion for factor analysis, (C) coding and statistical analysus, (D) factor analysis 39
Fig. 35. Monthly changes in fecundity of pomfrets 40
Fig. 36. Relationship among total length (TL), fork length (FL), ovary weight (OW), body weight (BW), oocyte diameter (OD) and fecundity. (A) TL, (B) FL, (C) OW,... 40
Fig. 37. Changes in survival rates for broodstock during wintering 41
Fig. 38. Observation of ovarian development and measurement of oocyte diameter for broodstock during wintering. (A~B) external feature of ovary,... 42
Fig. 39. Histological observation of (A) ovarian and (B) testicular development for P. argenteus under indoor tank wintering 42
Fig. 40. Histological observation of ovarian developmental process during induction of spawning for broodstock. (A) early growing stage with perinuclues stage oocytes,... 43
Fig. 41. External gonopore and gonadal features of P. argenteus for artificial fertilization 45
Fig. 42. Matured female (left), number of spawning, spawning time and volume of spawned eggs 46
Fig. 43. Changes in egg (A) and oil droplet (B) diameter of naturally spawned eggs from indoor reared broodstock 47
Fig. 44. Comparison of fertilization rate, hatching rate according to fertilization time. (A) comparison of fertilizated eggs by fertilization time, (B) release of polar body,... 49
Fig. 45. Comparison of changes in water temperature and mortality of pomfrets broodstock during wintering. (A) daily variation of water temperature and mortality,... 50
Fig. 46. Process of egg and sperm collection with wild stock and microscopic observation. (A) egg collection, (B) sperm collection, (C) fertilization, (D) egg washing,... 50
Fig. 47. Unfertilized egg and abnormal embryonic development of P. argenteus. (A) not ovulated egg, (B) paused development after fertilization, (C) paused development,... 52
Fig. 48. Embyro development of P. argenteus. (A) unfertilized egg, (B) fertilized egg (5 min. after fertilization), (C) 2 cells stage (0.5 HPF), (D) 128 cells stage (6 HPF),... 53
Fig. 49. Frequency analysis of fertilized egg diameter for pomfrets 54
Fig. 50. Comparison of fertilized eggs by artificial egg collection. (A) trasnportation of fertilized eggs, (B) collection of ripe eggs, (C) floated eggs after fertilization 59
Fig. 51. Feed regime and tank management used during the larval rearing of P. argenteus 60
Fig. 52. Morphological changes in hatched larvae of P. argenteus. (A) 2 days after hatching (DAH), (B) 6 DAH, (C) 12 DAH, (D) 14 DAH 60
Fig. 53. Changes in survival rates of P. argenteus hatched larvae during rearing period 61
Fig. 54. Comparison of (A) egg and (B) oil droplet diameter between natural and artificial fertilization 61
Fig. 55. Embryo developmental process of P. argenteus 62
Fig. 56. Daily growth of hatched larvae. Scale bars = 1mm 62
Fig. 57. Weekly changes in total length, body height and optic vesicle of hatched larvae P.argenteus 62
Fig. 58. Observation of molformation in hatched larvae P. argenteus. (A) (B) (D), malformation of vertebrate; (C) expansion of swim bladder 63
Fig. 59. Weekly changes of nutrient salts in rearing water for hatched larvae P. argenteus 64
Fig. 60. A comparative experiment for survival rate of hatched larvae P. argenteus according to the air supply method. (A) no air supply tank, (B) air supply tank 66
Fig. 61. Comparison of (A) total length and (B) survival rate in hatched larvae P. argenteus according to different air supply treatment 66
Fig. 62. Changes in rearing water condition according to different air supply treatment 67
Fig. 63. Observation of food digestion after feeding of hatched larvae P. argenteus 67
Fig. 64. Observation of air-bubble inhalation and followed intestinal stenosis. left, normal; middle, air-bubble inhalation and suspected intestinal stenosis; right,... 67
Fig. 65. Changes in total nitrogen levels by water exchange 68
Fig. 66. Changes in (A) water temperature and (B) survival rates of P. argenteus by high water temperature treatment 69
Fig. 67. Experiment tank (1ton) for intermediate culture. (A) flow-through tank as a control, (B) RAS tank 70
Fig. 68. Comparison of (A) total length and (B) survival rate between flow-through and RAS intermediate culture 70