Hernández Cruz, C.M.

Iakovopoulos, G.

Jerez, Salvador

Martín, María Virginia

Papandroulakis, Nikos

Pérez, José Antonio

English

Papandroulakis, N. (Nikos)

Pérez, J.A. (José Antonio)

Jerez, S. (Salvador)

Martín, M.V. (María Virginia)

Repositorio Institucional Digital del IEO

Hatchery production of the greater amberjack (Seriola Dumerili) N. Papandroulakis, Tsalafouta, P. Anastasiadis, I. E. Papadakis, N. Mitrizakis (HCMR) C.M. Hernández Cruz, A. La Barbera, H. Fernández-Palacios Barber, M. Izquierdo (FCPCT) S. Jerez, M. Virginia Martín (IEO) J. Perez, C. Rodríguez (ULL)  G. Iakovopoulos (GMF S.A.) Hellenic Centre for Marine Research Institute of Marine Biology, Biotechnology & Aquaculture n  Data on applicable rearing conditions in captivity is scarce ¨  particularly for the early developmental stages Aim: n  Define (some of) the parameters related to the larval rearing in order to optimize the applied methodologies ¨  larvae are optical predators ¨  adequate foraging of appropriate preys n  Objectives: to obtain new information on the larval rearing parameters  ¨ Acquire basic biological information  n  Ontogeny of digestion and vision system  n  Ontogeny of Somatotropic axis n  Skeletal Ontogeny ¨ Feeding regime - Prey enriching diet ¨ Husbandry parameters  n  tank type-shape,  n  stocking density,  n  light conditions  Ontogenesis of key biological systems Mesocosm	 Intensive	rearing	n  The	“pseudo-green”	water	methodology	–	closed	hydraulic	circuit	n  Controlled	conditions	¨  temperature	¨  photophase	¨  feed	distribution				n  computer	based	application	n  A	semi-intensive	technology	that	facilitates	the	rearing	of	several	species	providing	the	required	“biological	control”	Methodology (HCMR) Mesocosm n  Water: filtered (5 µm) natural seawater (salinity 40 psu) UV treated  n  T: 24 ± 0.7 °C n  pH: 8.0 to 8.2  n  D.O. 5.8 to 6.8 mg l-1.  n  Photophase: 24L:00D from 3 to 25 dph then to 18L:06D.  n  Light intensity: 500 lux to 1,000 lux,                               ~250 lux at night Intensive n  Water: filtered borehole 35 psu-water.  n  T:  22 ± 0.5°C until mouth opening             24 ± 0.5°C afterwards.  n  pH: 8.0 to 8.2  n  D.O. 6.8 to 7.2 mg l-1.  n  Photophase: 24L:00D from 3 to 25 dph then 18L:06D  n  Light intensity: 200 - 800 lux                              ~200 lux at night  n  Feeding ¨  3 to 18 dph : Microalgae (Chlorella sp) and enriched rotifers (Brachionus sp)  ¨  12 to 14 dph. Artemia AF A0 nauplii and 14 to 30 dph enriched Artemia EG A1 nauplii  ¨  16 dph (MES) and 21 dph (INT) artificial feeds (grain size 200–300 µm; and 300–500 µm)  ¨  Frozen gilthead sea bream eggs in MES after 20 dph ¨   Mesocosm tanks developed naturally zooplankton (harpacticoida copepods) ¨  Two different tanks types: 40,000 l and 2,000 l n  Photoperiod was natural (14:10 h, L:D),  n  S=37 psu, and  n  T=25-27ºC  n  DO=6.78 ± 0.5 ppm  n  Feeding: live phytoplankton (Nannochloropsis sp. ), enriched rotifers (1-30 dph), enriched Artemia (12-30 dph), microdiets of 75, 150 and 300 µm (13-30 dph) Methodology (FCPCT) Ontogeny of digestive and vision systems n  The digestive system  ¨  enables the fish to capture, ingest, digest and absorb nutrients from the food  ¨  is of special relevance to establish the larval  feeding protocol  ¨  ontogenesis of related organs and digestive enzymes 	The digestive system ontogenesis is a rapid process The gastric glands mark the transition from larval to juvenile and the development of a functional stomach •  in greater amberjack appeared after 5.5 mm of TL in all the rearing systems (16-20 dph).  Growth rate (exp) Survival Mesocosm 0.065 d-1 13.9±0.8% Intensive 0.047 d-1 8.2±3.1% Ontogenesis of digestive enzymes 0-3 dph: enzymatic activities before onset of exogenous feeding   importance of egg glycogen as energy during the embryonic development  3-5 dph: higher activity of alkaline protease than lipase   proteins the main energy source  5-10 dph: period of  rotifer feeding   proper nutritional condition of the larvae 10-15 dph: in both systems malnutrition   difficult acceptance of Artemia Ontogenesis of Vision system  –  0 dph: retina an undifferentiated neural epithelium (UNE)  –  1-2 dph the first structures appeared  –  ganglia cell layer (GCL), the inner plexiform layer (IPL), the inner nuclear layer (INL), the outer plexiform layer (OPL), the outer nuclear layer (ONL) and the photoreceptor layer (PL)).   –  3 dph- onwards: the pigment epithelium PE appeared on the external area of the retina –  the nucleus of the cone cells appeared in the outer nuclear layer –  Rods first appeared at TL 5.0±0.2 mm Lessons learned n  Transitions to juveniles after 5.5 mm n  Protein main energy source since first feeding  n  Night vision after 5.0 mm n  Rotifer appropriate as diet n  Difficult adaptation to Artemia Ontogenesis of the somatotropic axis IGF Binding Proteins 1, 2, 3, 4, 5 Available for the first time information on the regulation of the various components of the IGF signaling pathway in greater amberjack Skeletal Ontogeny TL: 3.38 ± 0.15mm Cranial development  TL: 4.49 ± 0.14mm Vertebral column and fin ontogeny TL: 4.49±0.14mm TL: 5.36±0.2mm TL: 13.03±0.09mm TL: 10.15 ± 1.86mm Completed bone mineralization of the vertebral column at 15mm Effect of feeding regime and probiotics n  Effect of  ¨  prey concentration and supply frequency  ¨  use of immune modulators substances during the rotifer administration n  Rotifers  ¨  Density: 5 vs 10 rot ml-1 ¨  Distribution 2 vs 3 times d-1 n  Four enriching diets ¨  A commercial diet (T1)  ¨  LC60/20:4n-6/10ppm carotenoids basic emulsion (T2),  ¨  LC60/20:4n-6/10ppm carotenoids basic emulsion with 20% Echium oil (T3)  ¨  LC60/20:4n-6/10ppm carotenoids basic emulsion with 20% black cumin oil (T4) Larval growth and survival ¨  Similar regardless rotifer density  n  slightly better at 5 rot ml-1 ¨  Better for  feeding 3 times day-1 ¨  Better with T4 Humoral innate immune activities 7 dph and 12 dph larvae  Effect of larvae density n  3 densities: 25, 50 and 75 eggs l-1 ¨  Two different tanks types: 40,000 l and 2,000 l tested for 30 days n  Photoperiod natural (14:10 h, L:D),  n  S: 37 psu,  n  T: 25-27ºC  n  DO: 6.78 ± 0.5 ppm  n  Feeding: live phytoplankton (Nannochloropsis sp. ), enriched rotifers (1-30 dph), enriched Artemia (12-30 dph), microdiets of 75, 150 and 300 µm (13-30 dph) 25Eggs/l 50Eggs/l 75Eggs/lSurvival (%)02468101214161825 eggs/l50 eggs/l75 eggs/lbabaDeformities observed A: Lordosis  B, G: Vertebral fusion  C: Fusion of neural arch and spines  D: Fusion of caudal vertebrae 	F,	H,	I:	Anomalous	dentary 	G:	Partial	vertebral	fusion	in	caudal	vertebrae 	J:	Cephalic	anomalies	(glossohyal)  Effect of environmental parameters Tank hydrodynamics ¨  Two different tanks types: 40,000 l and 2,000 l were tested n  eggs from natural spawning (10 eggs l-1)  n  photoperiod natural (14:10 h, L:D),  n  S: 37 psu,  n  T: 25-27ºC.  n  DO: 5–8 g l-1.  n  Feeding: live phytoplankton (Nannochloropsis sp. ), enriched rotifers (1-30 dph), enriched Artemia (12-30 dph), microdiets of 75, 150 and 300 µm (13-30 dph) 2,000	Lt40,000	LtDifferences in current profiles (generally higher in the 2,000 l tanks)   Survival low: 2-3% The effect of light (intensity and duration) •  Photophase 18L:06D vs 24L:00D)  •  Background color (black, green, white) Intensive	rearing	n  The “pseudo-green” water methodology – closed hydraulic circuit ¨  Water: filtered borehole 35 psu-water.  ¨  T:  22 ± 0.5°C until mouth opening                         24 ± 0.5°C afterwards.  ¨  pH: 8.0 to 8.2  ¨  D.O. 6.8 to 7.2 mg l-1.  ¨  Photophase: 24L:00D from 3 to 25 dph then 18L:06D  ¨  Light intensity: 200 - 800 lux                                           ~200 lux at night Growth rate (exp) Survival 18L:06D 0.031 d-1 10.6±4.2% 24L:00D 0.031 d-1 8.2±3.1% Results - Photophase      SURVIVAL: WHITE:    22,2%                          BLACK:      8,2%                         GREEN:    16,5% Growth rate (exp): White:  0.0393 d-1                                 Black:  0.0481 d-1                                Green:  0.0355 d-1  0	2	4	6	8	10	12	14	0	 5	 10	 15	 20	 25	TL	(mm)	Time	(days)	Black	White	Green	q  Different expression of genes IGF-I, IGF-II and IGF Binding Proteins 2,3 & 4  o  higher levels for WHITE background Results - Background color n  Max light intensity in the water column at 30 cm ~50 lux n  3-4x higher than without light Table 3. Measured light intensity (Lux) at several points in both tanks of each treatment. Values are means ± standard deviation. Measurement point Green-no light White-no light Green-light White-light Black Under surface Centre (n=2) 334 ± 4 303 ± 14  255 ± 11  234 ± 1.1 295 ± 24.6   Side  (n=8) 86 ± 102 78 ± 98 64 ± 10 66 ± 82 68 ±11 Depth ~30cm Angle 45° (n=8) 15 ± 7.4 13 ± 6.2 11 ± 4.5 11 ± 7.7 12 ± 2   Towards centre (n=8) 11 ± 1.7 a 12 ± 1.1 a 31 ± 1.7 b 27 ± 4.8 b 9 ± 3 n  This study showed a catalytic effect of the light conditions on larval survival ¨  may reflect requirements related to the pelagic nature of the species  n  The presented results with the modified “light environment” are better at one order of magnitude of any previous reported  n  Indicate a substantial technological step in the larval rearing of the greater amberjack   Towards an industrial protocol  Validation in Commercial hatcheries n  Hatchery 1  ¨  Larvae density 75 ind l-1 ¨  Light intensity n  800 lux on 3-6 dph, 1200 lux on 6-12 dph 1000 lux 15 dph 500 lux 20 dph  ¨  Photophase: 24L:00D from 3-20 dph then 18L:06D  ¨  Feeding: phytoplankton, enriched rotifers, Artemia and dry feeds. Frozen eggs after 20dph ¨  Size selection during weaning  ¨  In 2017 n  15.000 juveniles classified in 4 size-classes between 0.3 and 2.5 g to nursery ¨  In 2018 similar method n  until 20 dph survival ~10% n  between 20-30 dph: no sorting            mortality >65% due to cannibalism  n  30 dph 15.000 individuals were transferred to nursery n  Hatchery 2 ¨  Direct incubation of eggs in the larval rearing tanks  ¨  T: 24.5 to 25.0 ºC.  ¨  pH: 7.8 ¨  Light conditions : modified in some tanks n   intensity on the surface >1000 lux ¨  Photophase: 24L:00D from 3-20 dph then 18L:06D  ¨  Feeding: enriched rotifers, instar I (TL >5mm) and enriched instar II (TL >6mm) Artemia nauplii and artificial diets (TL >7mm).  ¨  Tanks with high light intensity performed better ¨  Sorting in size from 20 dph, improves significantly survival  n  48.300 juveniles of 25-50 g transferred in cages  Validation in Commercial hatcheries (2) Recommendations (1)  n  Rearing parameters ¨  large tank improves growth performance and survival ¨  egg stocking densities ~ 25 eggs l-1 ¨  DO: 5.0 to 8. mg l-1, preferably >6.0 mg l-1  ¨  S: 37 - 40 psu  ¨  pH: 7.8-8.5  ¨  T: 22 to 27ºC, preferably between 23.5 and 25.0ºC ¨  Photophase 18L:06D from 1 to 25 dph  ¨  Light intensity: Defused light of 800-1200lux, 3-12 dph                                         gradually 800 lux until 20 dph n  Feeding ¨  coordinate rearing conditions and larval development  ¨  live microalgae at 150-300 x 103 cell ml-1 since 2 dph  ¨  enriched rotifers min 2x d-1, 3-15 dph, at 3-7 rot ml-1; Artemia AF (TL>5mm), for 2-3 days; enriched Instar II (TL> 6.0 mm); dry diet (200-800 µm) ) (TL>6.5 mm) ¨  immune modulators (Echium and black cumin oil) together with optimized enriching emulsion and astaxanthin as carotenoid improve larval performance n  Husbandry ¨  high size variability (at 20-30 dph) in all rearing systems tested until today (unknown reasons)  n  early sorting to appropriate size classes  n  standard methods and equipment available in all hatcheries  n  significant higher survival compared to unsorted groups although the losses  n  mortality for unsorted groups >90%, for sorted groups 10-15%. ¨  Transportation of individuals n  larvae with TL <15 mm do not tolerate netting, transfer performed with care.  n  larvae with TL >20 mm can be netted normally  ¨  individuals >0.5-1 gr are handled easier  n   light anesthesia may help Recommendations (2)  2mm 5mm 20mm Thank you! Hellenic Centre for Marine Research  Institute of Marine Biology, Biotechnology & Aquaculture 

Hatchery production of the greater amberjack (Seriola dumerili)

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http://www.repositorio.ieo.es/e-ieo/bitstream/10508/12317/1/papandroulakis_n_hatchery_production%20%281%29.pdf

Hatchery production of the greater amberjack (Seriola dumerili)

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