The effect of temperature on the duration of spawning markers—migratory-nucleus and hydrated oocytes and postovulatory follicles—in the multiple-batch spawner Japanese flounder (Paralichthys olivaceus)

The duration of spawning markers (e.g. signs of previous or imminent spawnings) is essential information for estimating spawning frequency of fish. In this study, the effect of temperature on the duration of spawning markers (i.e., oocytes at early migratory nucleus, late migratory nucleus, and hydrated stages, as well as new postovulatory follicles) of an indeterminate multiple-batch spawner, Japanese f lounder (Paralichthys olivaceus), was evaluated. Cannulation was performed to remove samples of oocytes, eggs, and postovulatory follicles in individual females at 2–4 hour intervals over 27–48 hours. The duration of spawning markers was successfully evaluated in 14 trials ranging between 9.2° and 22.6°C for six females (total length 484–730 mm). The durations of spawning markers decreased exponentially with temperature and were seen to decrease by a factor of 0.16, 0.36, 0.30, and 0.31 as temperature increased by 10°C for oocytes at early migratory nucleus, late migratory nucleus, and hydrated stages, and new postovulatory follicles, respectively. Thus, temperature should be considered when estimating spawning frequency from these spawning markers, especially for those fish that do not spawn synchronously in the population

Aquaculture Asia, Vol. 11, No. 1, pp.1-36, January - March 2006

*Table of Contents* Research & farming techniques Nursery rearing of Puntius goniotus: A preliminary trial K.N. Mohnta, J.K. Jena & S.N. Mohanty Artemia enrichment and biomass production for larval finfish and shellfish culture A.S. Ninawe Vembanad Lake: A potential spawner bank of the giant freshwater prawn Macrobrachium rosenbergii on the southwest coast of India Paramaraj Balamurugan, Pitchaimuthu Mariappan & Chellam Balasundaram Seed production of mud crab Scylla serrata at the Rajiv Gandhi Center for Aquaculture, Tamil Nadu, India Mohamed Shaji, Emilia T. Quinitio, Thampi Samraj, S. Kandan, K. Ganesh, Dinesh Kumar, S. Arulraj, S. Pandiarajan, Shajina Ismail and K. Dhandapan. Sustainable aquaculture Fish wastes in urban and suburban markets of Kolkata: Problems and potentials Kausik Mondal, Anilava Kaviraj & P.K. Mukhopadhyay People in aquaculture Peter Edwards writes on rural aquaculture: Farming carps in leased ponds by groups of poor women in Chandpur, Bangladesh Aquatic animal health Lymphocystis disease and diagnostic methods in China Jing Xing, Xiuzhen Sheng & Wenbin Zhan Asia-Pacific Marine Finfish Aquaculture Network Mesocosm technology advances grouper culture in northern Australia Elizabeth Cox, Peter Fry & Anjanette Johnsto

ヒラメの獲得免疫制御に及ぼすサイトカインの影響

東京海洋大学博士学位論文 平成25年度(2013) 応用生命科学 課程博士 甲第304号指導教員: 近藤秀裕全文公表年月日: 2018-05-17東京海洋大学201

Effect of feeding ratio on growth and body composition of juvenile and sub-adult olive flounder Paralichthys olivaceus

여름철 동안 부상용 배합사료 (Extruded pellet)의 다양한 사료 공급율에 따른 넙치 (Paralichthys olivaceus) 유어기 (실험 Ⅰ) 및 미성어기 (실험 Ⅱ)의 성장과 체조성에 미치는 영향을 조사하였다 . 실험 Ⅰ에는 21개의 180 L 유수식 탱크에 30마리의 유어 (시작 시 무게 17 g)를 수용하였다 . 실험에 이용된 사료 공급율은 5% 간격으로 설정하였고 , 총 실험구는 7구간 (100% 만복 , 만복의 95%,90%, 85%, 80%, 75% 및 70%)을 두었으며 , 각 실험구는 3 반복구로 7주 동안 사육실험을 실시하였다 . 실험 Ⅱ에는 15개의 500 L 유수식 탱크에 13마리의 미성어 (시작 시 무게 319 g)를 수용하였다 . 실험에 이용된 사료 공급율은 5% 간격으로 설정하였고 , 총 실험구는 5구간 (100% 만복 , 95%, 90%, 85% 및 80%)을 두었으며 , 각 실험구는 3반복구로 10주 동안 사육실험을 실시하였다 . 실험Ⅰ과 Ⅱ에서의 사료 공급양은 대조구 (100% 만복 )에서는 1일 2회 매일 손으로 만복시 까지 사료를 공급하였다 . 나머지 6구간 (실험 Ⅰ) 및 4구간 (실험 Ⅱ)은 대조구의 일일 사료 섭취량에 따라서 매일 조절하여 공급하였다 . 실험 Ⅰ에서 유어 넙치의 생존율은 사료 공급율에 따른 유의적인 차이를 보이지 않았다 (P>0.05). 사료를 만복시 까지 공급한 대조구의 어체중 증가 및 일일성장률 (SGR)은 만복의 95%를 공급한 실험구와 유의적인 차이를 보이지 않았지만 , 만복의 90%, 85%, 80%, 75% 및 70%를 공급한 실험구보다 어체중 증가 및 일일성장률은 유의적으로 높게 나타났다 (P0.05). 어체와 간의 조단백질 함량은 유의적인 차이를 보였지만 (P0.05). 간체장지수 (HSI)와 비만도 (CF)는 사료 공급율에 따른 유의적 차이를 보이지 않았다 (P>0.05). 실험 Ⅱ에서 사료를 만복시 까지 공급한 미성어 넙치의 어체중 증가는 만복의 85%와 80%를 공급한 실험구와 유의적인 차이를 보였지만 (P<0.05), 만복의 95%와 90%를 공급한 실험구와는 유의적인 차이를 보이지 않았다 (P>0.05). 사료효율 (FER), 단백질전환효율 (PER) 및 단백질축적율 (PR)은 사료공급율에 따른 유의적인 차이를 보이지 않았다 (P>0.05). 사료 공급율에 따른혈청의 total protein, glucose 및 glutamic pyruvic transaminase는 유의적인 차이를 보이지 않았지만 , triglyceride와 glutamic oxaloacetic transaminase는 유의적인 차이를 보였다 (P<0.05). 이상의 결과를 고려할 때 , 여름철 유어기 넙치의 적정 사료 공급율은 만복의 95%이고 , 미성어기는 90%로 판단되며 , 넙치가 성장함에 따라서 적정 사료 공급율은 감소하였다 .The effect of feeding ratio on growth and body composition of juvenile (feeding trial Ⅰ) and sub-adult (feeding trial Ⅱ) olive flounder (Paralichthys olivaceus) fed extruded pellets was determined during the summer season. In the feeding trial Ⅰ, thirty juvenile olive flounder (initial body weight of 17 g) per tank were distributed into 21, 180 L flow-through tanks. Seven treatments that included triplicate groups of feeding ratio in 5% decrement were prepared for this study: 100% (satiation), 95%, 90%, 85%, 80%, 75% and 70% of satiation. In the feeding trial Ⅱ, thirteen sub-adult olive flounder (an initial body weight of 319 g) per tank were distributed into 15, 500 L flow-through tanks. Five treatments of feeding ratio in 5% decrement were prepared in triplicate: 100% (satiation), 95%, 90%, 85% and 80% of satiation. Fish in the control group (100% of satiation) were hand-fed to apparent satiation twice a day for 7 and 10 weeks in the feeding trials Ⅰ and Ⅱ, respectively. Then feed allowance in the rest of six for juvenile and four groups for sub-adult was determined based on average feed consumption of fish in the control group. In the feeding trial Ⅰ, survival of juvenile olive flounder was over 97% and was not significantly (P>0.05) affected by the feeding ratios. Weight gain and specific growth rate (SGR) of fish fed to 100% of satiation were not significantly different from those fed to 95% of satiation but significantly (P0.05) affected by the feeding ratio. The crude protein levels of the whole body without liver or liver were significantly affected by the feeding ratio. Hepatosomatic index and condition factor of fish were not significantly (P>0.05) affected by the feeding ratio. In the feeding trial Ⅱ, weight gain of sub-adult olive flounder fed to 100% of satiation was significantly (P0.05) different from that of fish fed to 95% and 90% of satiation. However, efficiency (FER, PER, and PR) of diet of flounder was not significantly (P>0.05) affected by feeding ratio. Serum total protein, glucose and glutamic pyruvic transaminase were not significantly (P>0.05) affected by feeding ratio, but triglyceride and glutamic oxaloacetic transaminase were. In considering the results from the feeding trials Ⅰ and Ⅱ, it can be concluded that optimum feeding ratio for growth of juvenile and sub-adult olive flounder could be lowered to 95% and 90% of satiation, respectively, without growth suppression when fish were fed the extruded pellets twice a day during the summer season. And optimum feed allowance decreased as fish grew.General Introduction = 1 Feeding trial Ⅰ. Effect of feeding ratio on growth and body composition of juvenile olive flounder paralichthys olivaceus fed extruded pellets during the summer season = 3 Abstract = 3 1. Introduction = 3 2. Materials and Methods = 4 2. 1. Fish and the experimental conditions = 4 2. 2. Design of the feeding trial = 4 2. 3. Preparation of the experimental diet = 4 2. 4. Analytical procedures of fish = 5 2. 5. Statistical analysis = 5 3. Results = 5 4. Discussion = 6 5. References = 8 Feeding trial Ⅱ. Effect of daily feeding ratio on growth and body composition of sub-adult olive flounder,Paralichthys olivaceus, fed extruded pellet during the summer season = 10 Abstract = 10 1. Introduction = 11 2. Materials and Methods = 12 2. 1. Fish and the experimental conditions = 12 2. 2. Design of the feeding trial = 12 2. 3. Preparation of the experimental diet = 13 2. 4. Analytical procedures of fish = 13 2. 5. Blood analysis of fish = 13 2. 6. Statistical analysis = 15 3. Results and Discussion = 15 4. Literature Cited = 24 Conclusion = 29 국문요약 = 30 Acknowledgments = 32 References = 3

Prevention of hypermelanosis by rearing Japanese flounder Paralichthys olivaceus in net-lined tanks

In artificially reared flatfish, especially the Japanese flounder Paralichthys olivaceus, pigmented skin (hypermelanosis) frequently appears on the fish’s blind side after normal metamorphosis. As no practical prevention method has yet been proposed, we examined the effectiveness of a loose net placed inside the rearing tank that covers the bottom and walls like a pouch. When juveniles (standard length [SL] 6 cm) were transferred to the net-lined tank (mesh size 4 mm) before the first appearance of hypermelanosis, the pigmented area after 2 months covered about 0.5% of the blind side; this is about 1/40th of the area covered by pigment in fish reared in an ordinary tank (20%). Although the initial appearance of pigmentation in the axilla area (the area covered by the pectoral fin) was not suppressed, utilization of a larger mesh size (12 mm) decreased the expansion of pigmentation in this area. Juveniles reared in the net-lined tank were about 5–15% smaller (SL) than those reared in the ordinary tank, but their body depth:SL ratio was closer to that of wild-caught juveniles. From the results of this study, we propose that net-lined rearing tanks with larger-sized mesh are a practical method of preventing hypermelanosis in Japanese flounder aquaculture systems

Effects of time and duration of rearing with bottom sand on the occurrence and expansion of staining-type hypermelanosis in the Japanese flounder Paralichthys olivaceus

We previously reported that the progression of staining-type hypermelanosis spontaneously ceased at a specific time and area in Japanese flounder Paralichthys olivaceus. To examine whether time is a limiting factor in the spontaneous cessation of staining, we experimentally controlled the initiation and duration of staining by manipulating the bottom substrate condition in the fish tanks. At 151 days post hatching (DPH; 11 weeks), spontaneous cessation of staining was observed in fish reared in tanks without a sandy substrate. However, staining resumed (or was initiated) in tanks where sand was removed from 11 weeks, indicating a strong but temporary effect of bottom sand and the absence of time limitation in the staining progression by 151 DPH. Extended duration of the inhibitory period of hypermelanosis expansion (9 weeks or more) aided in only a 20 % reduction of the final staining area because of the increased rate of staining expansion. The bottom sandy substrate decreased the visibility of the staining area in individuals, but this was observed only before the completion of the staining expansion. These findings are discussed in relation to possible presence of area limitation of future staining, as well as the fundamental nature of staining

Response of dietary substitution of fishmeal with various protein sources on growth, body composition and blood chemistry of olive flounder (paralichthys olivaceus)

Animal and/or plant protein sources substituting fishmeal in the diets keep being developed due to its high price. The purpose of this study is to determine response of dietary substitution of fishmeal with silkworm pupae meal, promate meal, meat and bone meal and/or their combination on performance of juvenile olive flounder (Paralichthys olivaceus). A 60% fish meal was used as the main protein source, used as the control (Con) diet. The 10 and 20% fishmeal were substituted with silkworm pupae meal and meat and bone meal, referred to as the SPM10, SPM20, MBM10 and MBM20 diets, respectively. And the 10, 20 and 40% fishmeal were substituted with promate meal, referred to as the PM10, PM20 and PM40 diets, respectively. Finally the 10 and 20% fishmeal were substituted with combined silkworm pupae meal and promate meal, refereed to as the SPM+PM10 and SPM+PM20 diets, respectively. Weight gain and specific growth rate of fish fed the MBM10 diet were higher than those of fish fed the Con, SPM20, PM20, PM40 and SPM+PM20 diets. Feed efficiency ratio of fish fed the SPM10, MBM10, MBM20, PM10 and SPM+PM10 diets was higher than that of fish fed the SPM+PM20 and PM40 diets. Protein efficiency ratio of fish fed the MBM10 and MBM20 diets was higher than that of fish fed the SPM20, PM20, PM40 and SPM+PM20 diets. In conclusion, dietary substitution of fishmeal with 10% SPM, 20% MBM, 10% PM and 10% SPM+PM could be made.Contents i List of Tables ii Abstract (in Korean) iii Abstract v Abbreviation vii I.Introduction 1 II. Materials and methods 3 2.1. Fish and the Experimental Conditions 3 2.2. Design of the Feeding Trial and Preparation of the Experimental Diets 3 2.3. Analytical Procedures of the Experimental Diets and Fish 7 2.4. Statistical Analysis 8 III. Results and discussion 9 IV. Conclusions 22 V. Acknowledgements 23 VI. References 2

Protein requirement in granulated microdiets for olive flounder (Paralichthys olivaceus) larvae

본 연구에서는 넙치 자치어의 과립형 미립자 초기사료내 적정 단백질 요구량에 대하여 조사하였다. 실험사료는 조단백질 함량이 42%에서부터 58%로서 4%씩 증가시키는 5종류의 과립형 미립자 초기사료(CP42, CP46, CP50, CP54, CP58사료)를 준비하였으며, 이때 에너지 함량은 4.42 kcal g-1 diet로 일정하게 유지시켰다. 실험사료를 공급하는 실험구는 3반복구를 두었다. 부화 후(DAH) 14일령의 넙치 자치어 7,500미를 무작위로 추출하여 15개의 70 L 사각형플라스틱 탱크에 탱크당 500마리씩 각각 수용하였다. 과립형 미립자 초기사료의 단백질 함량이 증가함에 따라 모든 필수아미노산(AA) 및 비필수아미노산의 함량은 증가하였다. 사육실험 종료시 넙치 자치어의 생존율은 실험사료에 따른 유의적인 차이를 보이지 않았다. 그러나 넙치 자치어의 마리당 증체량(weight gain)과 성장률(growth rate)은 CP54와 CP58사료 공급구에서 CP42, CP46와 CP50사료 공급구보다 유의적으로 높게 나타났다. 또한 자치어의 전장(total length)은 CP54와 CP58사료 공급구에서 CP42와 CP46사료 공급구보다 유의적으로 길게 나타났다. 넙치 자치어의 조지방 함량은 CP58사료 공급구에서 CP42, CP46와 CP50사료 공급구보다 유의적으로 높았다. 사육 실험 종료시 생존한 어체의 체구성 아미노산 분석 결과, 실험사료의 종류에 따른 유의적인 차이는 보이지 않았다. 이상의 결과를 고려할 때, 넙치 자치어의 과립형 미립자 초기사료내 단백질 요구량은 성장률(growth rate)에 근거하면 55.2%인 것으로 평가된다. | The optimal protein requirements in granulated microdiets were determined for larval olive flounder. Five granulated microdiets (CP42, CP46, CP50, CP54 and CP58), containing different levels of protein ranging from 42 to 58% and at a constant estimated energy level (4.42 kcal g-1 diet),were prepared in triplicate. 14 days after hatching (DAH), 7500 larvae were placed in 15 indoor 70 L square plastic tanks. As the protein levels increased in the granulated microdiets, all essential and nonessential amino acid (AA) contents increased. The weight gain and growth rate of the flounderlarvae fed the CP54 and CP58 diets were greater than of larvae fed the other (CP42, CP46 and CP50) diets. The total length of larval flounder fed the CP54 and CP58 diets was greater than of the flounder fed the CP42 and CP46 diets. The crude lipid content of the larval flounder fed the CP58 diet was higher than of the flounder fed the CP42, CP46 and CP50 diets. None of the whole body AA profiles of the founder larvae was affected by the protein levels in the granulated microdiets. In conclusion, dietary protein requirement was estimated to be 55.2% based on the growth rate of the larval olive flounder.I. Experiment 1 Abstract 1 1. Introduction 3 2. Materials and Methods 6 2.1. Spawning and larval rearing conditions 6 2.2. Preparation of the experimental diets 6 2.3. Experimental conditions 10 2.4. Analytical procedures for the microdiets and larval fish 11 2.5. Statistical analysis 11 3. Results 13 4. Discussion 19 II. Conclusion 23 III. Acknowledgements 24 IV. References 25Maste

THE EFFECTS OF TEMPERATURE ON CYP19A1A, FOXL2, DMRT1 AND AMH EXPRESSION DURING SEX DIFFERENTIATION IN SUMMER FLOUNDER (PARALICHTHYS DENTATUS)

Female summer flounder grow considerably faster and larger than males, and a tremendous increase in performance can therefore be realized through production of monosex female populations. Rearing temperature has been shown to affect sex differentiation in other teleost species by influencing expression of genes encoding transcription factors, hormones or enzymes involved in endocrine function such as cyp19a1a, foxl2, dmrt1 and amh. These genes have been linked to female (cyp19a1a, foxl2) or male (dmrt1, amh) development, and exhibit sexually dimorphic expression in some species. In the present study, summer flounder (37 days post hatch; DPH) were raised at 13°C, 16°C or 19°C. Fish from all three treatments were sampled during early development (38-66 DPH), and fish from 13 and 16°C treatments were sampled through later juvenile development (191 DPH). A partial summer flounder cDNA sequence for cyp19a1a was identified, verified, and submitted to the NCBI GenBank database. Partial summer flounder sequences were also were identified for foxl2, dmrt1 and amh, but these single sequence reads were not verified or submitted to NCBI. Summer flounder samples were analyzed in qPCR to determine cyp19a1a, foxl2, dmrt1 and amh gene expression levels. Sex ratios of additional fish grown to \u3e 150 mm at each temperature treatment were determined. Low female production was achieved overall (26.9, 17.6 and 0% at 13, 16 and 19°C, respectively). Cyp19a1a expression was significantly lower at 52 DPH (~15 mm total length) at the male producing temperature (19°C), and increased to similar levels as other treatments at 66 DPH. Cyp19a1a expression levels later in juvenile development (66-191 DPH) largely decreased with fish size. No clear trend in gene expression levels was present for foxl2, dmrt1 or amh. The period of sex differentiation in summer flounder remains unknown but cyp19a1a expression patterns suggest that it may occur earlier in development than that of congenerics, and that cyp19a1a may be a more robust sex marker in summer flounder than foxl2, dmrt1 or amh. Further research is necessary to understand the sex-determining mechanisms in this species before sexually dimorphic growth can be used to achieve economic advantages in commercial production