Recent developments in the application of live feeds in the freshwater ornamental fish culture

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Recent developments in the application of live feeds in the freshwater ornamental fish culture Lian Chuan LIM, Agri-food & Veterinary Authority of Singapore Philippe Dhert, INVE Technologies NV, Belgium Patrick Sorgeloos, Ghent University, Belgium

Ornamental Fish Business Netherlands 2.8% Philippines 3.9% Others 26.5% Singapore 26.4% Israel 4.3% Japan 4.6% Sri Lanka 5.0% Malaysia 5.3% Czech Rep 6.4% USA 6.5% China 8.2% Multi-million-dollar business Total export trade in 1998: US$ 163 mil.

Due to lack of research input and technology promotion, the freshwater ornamental fish culture technology lags far behind that of marine foodfish, especially in larviculture

Live Feeds for Fry Production Marine foodfish species: Availability of large quantity of live feeds has contributed to its successful fry production Freshwater ornamental fish culture: Industrial development has been hampered by the lack of suitable live feeds for feeding

Traditional Feeds for Freshwater Ornamental Fish Culture For larval feeding Inert food items: Egg yolk suspension, milk powder or powdered feeds Natural plankton bloom induced by pond fertilization. Feeding of larger fish and brooders Moina & Tubifex cultured in water fertilized with organic manure

Problems of using traditional feeds limits the fish stocking density due to poor water quality adversely affects fish quality, e.g. parasitic infection no suitable live feed for feeding larvae with small month size

Objective To explore the application of four live feeds in freshwater ornamental fish culture: Freshwater rotifers, Brachionus calyciflorus Artemia nauplii Decapsulated Artemia cysts On-grown Artemia

Use of rotifers Marine rotifers (Brachionus plicatilis) can survive in fresh water for at least 2 h sink quickly to bottom - not suitable for feeding pelagic larvae More appropriate to use freshwater rotifers, e.g. B. calyciflorus

Use of Artemia nauplii Major drawback in feeding Artemia nauplii to freshwater fish: Nauplii die in fresh water within an hour Can be overcome by cold storage nauplii at 4 o C. The technique would allow a constant supply of high quality nauplii, more frequent feeding to FW fish larvae and fry

Dwarf Gourami (Colisa lalia) Larvae measure 2.74 mm Raised in fertilized ponds of >100 m 3 capacity Fed natural plankton supplemented with egg yolk particles for first 10 days Low stocking density of <0.5/l

Dwarf Gourami: Effects of rotifer feeding Phase 1: Rotifer vs egg-yolk feeding, 30 larvae/l Phase 2: Both groups fed Artemia, 10 larvae/l Total length (mm) 12 10 8 6 4 2 Egg-yolk group Rotifer group b a a b Survival (%) 100 80 60 40 20 a b Egg-yolk group a Rotifer group b 0 Rotifer (D12) Artemia (D32) 0 Rotifer (D2-12) Artemia (D13-32) Feeding phase Feeding phase Food is not limiting, quality of feed in initial feeding is crucial to later developments

Extensive culture in ponds using egg-yolk Intensive culture in tanks using rotifers Overall survival (%) 17.5% 65.1-74.5% Yield (No./m 3 ) 90 6,500 7,500 Use of rotifers for feeding would enable intensive larviculture and improve the larval performance in freshwater ornamental fish.

Discus: Parental feeding Feed on body mucus of parent fish during first two weeks Problems: Risk of larvae being eaten up Parent fish do not spawn

Discus: Artificial feeding Smuggle valuable larvae to low quality parents (foster parents) : Risk of cannibalism Smear yolk-food below water line: change water and food prepared afresh; tedious and laborious

Discus larvae could be raised in the absence of the parent fish through feeding with B. calyciflorus from D4-7 Freshwater rotifers Artemia nauplii for a week, D8-14 Artemia nauplii

Growth and survival were comparable to parental feeding Total length (mm) 12 10 8 6 4 2 0 AF30 AF20 PF20 Rotifer (D7) NM Artemia (D14) Survival rate (%) 100 80 60 40 20 0 AF30 AF20 PF20 Rotifer (D2-7) Artemia (D8-14) Feeding phase Feeding phase AF30: Artificial feeding; 30 larvae/l; AF20: Artificial feeding; 20 larvae/l PF30 : Parental feeding; 30 larvae/l; NM: Not measured

Discus: Advantages of using live feeds for artificial feeding Safe: no cannibalism Less tedious: frequent change of water not required More practical for commercial application

Use of Artemia nauplii in Guppy Culture: Stress resistance Performances of Guppy adults and fry fed Artemia nauplii are better than those fed Moina 300 250 Artem ia Moina a a a a Stress index 200 150 100 50 0 Fry Adults

Use of Artemia nauplii in Guppy Culture: Growth Wet weight 350 300 250 200 150 100 Artem ia Moina a b a b 50 0 Fry Adults

Use of decapsulated cysts Have been used for feeding larvae and fry of marine shrimp freshwater prawn freshwater foodfish Their use in marine fish not successful, as the larvae and fry do not have the necessary enzyme for digestion of the cysts embryonic membranes.

Guppy Adults Parameters Stress index (at 35 ) Wet weight (mg) Brine cysts 225.5 a (4.80) 323.3 a (19.06) Dried cysts 242.8 a (9.91) 343.9 a (15.48) Artemia nauplii 233.5 a (17.75) 323.1 a (14.43) Moina 246.5 a (7.33) 260.3 b (15.76) Fish fed on cyst diets: No difference in stress resistance with AN and MN Dry weight (mg) Total length (mm) Survival rate (% ) 112.6 a (10.97) 30.2 a (0.25) 100.0 a (0) 119.5 a (6.12) 31.1 a (0.53) 99.8 a (0.50) 110.1 a (2.62) 30.2 a (0.52) 99.5 a (1.00) 78.0 b (2.44) 29.0 b (0.62) 98.0 b (0.82) No difference in growth & survival with AN Better growth and survival than MN

Guppy fry Parameters Stress index (at 30 ) Wet weight (mg) Dry weight (mg) Total length (mm) Survival rate (%) Brine cysts 210.5 a (3.11) 92.0 b (1.04) 24.9 b (1.54) 20.6 b (0.47) 65.3 a (4.33) Dried cysts 221.8 b (5.25) 122.3 a (4.94) 37.0 a (2.87) 21.7 a (0.46) 60.6 a,b (1.11) Artemia nauplii 247.0 c (6.88) 91.9 b (8.51) 23.9 b,c (2.51) 20.2 b (0.51) 58.0 b (2.71) Moina 253.5 c (4.51) 76.7 c (4.71) 19.1 c (2.58) 20.1 b (0.66) 57.0 b (3.72) Fry fed on cysts diets displayed: Better stress resistance than AN & MN Better growth than MN; better or comparable growth with AN Better or comparable survival with AN or MN

For Guppy fry and adults fed cysts diet, their performance in terms of stress resistance, growth and survival are better than or comparable with those fed Artemia nauplii and Moina Similar results were obtained in the fry of Molly (Poecilla sphenops) Platy (Xiphophorus maculatus) Swordtail (X. helleri) Black Neon Tetra (Hyphessobrycon herbertaxelrodi)

Better performance in cyst-fed fish could at least partly due to superior fatty acid composition of cysts Corresponds to higher energy content in Artemia cysts than nauplii Fatty acids (mg/g DW) Brine cysts Dried cysts Artemia naplii Moina 20:5(n-3) EPA 4.0 4.4 0.9 2.3 22:6(n-3) DHA 0.7 1.9 0.3 0.2 Total (n-3)hufa 6.0 8.0 2.0 3.1 DHA/EPA ratio 0.17 0.44 0.35 0.10 (n-6)/(n-3) 0.21 0.22 0.26 0.84 Total FAME 109.1 126.4 47.5 73.2

Decapsulated Artemia cysts can be used as a substitute for Artemia nauplii or Moina Advantages Being a hygienic off-the-shelf feed Direct use of the cysts signifies a new area of application of low-hatch cysts in the OF industry Saving in feed cost

Use of on-grown Artemia Bigger and older on-grown Artemia may be a good alternative live feed for feeding to larger OF including brooders Its use not as popular as Artemia nauplii, due to lack of supply Culture system developed in Singapore to facilitate supply for OF culture

Pilot production system for On-grown Artemia 21 culture raceways (5.6 m 3 each); each unit consists of 18 air-lift pumps and 2 waste collectors

Pilot production system for On-grown Artemia Uses artificial seawater of 20 for culture Artemia are fed rice bran and de-fat soybeans Production rate: 3 kg/m 3 in 12-day cycle Production capacity: 8 mt./yr Construction cost: US$ 82,000 Compared with existing system: cost effective, simple and easy to set up and operate

Size of on-grown Artemia Total length (mm) 5 4 3 2 1 0 0 2 4 6 8 10 12 Days of culture Grew from 0.45 mm to 5mm in 12 days - size range suitable for feeding all ornamental fish up to 10 cm TL. By varying harvesting time, possible to tailor prey size accordingly to size and age of fish - other live feeds such as rotifers and Artemia nauplii do not have such flexibility

Feeding of on-grown Artemia to Discus juveniles Due to more effective food uptake, the Artemia-fed group grew faster than fish fed Moina or frozen bloodworms Survival: no significant difference Fish feeds Parameters On-grown Artemia Moina Frozen bloodworms W et weight (g) 0.85 a (0.01) 0.81 b (0.01) 0.75 c (0.01) Total length (mm) 3.48 a (0.03) 3.44 a,b (0.04) 3.37 b (0.04) Survival rate (%) 90.0 a (13.2) 78.3 a (10.4) 91.7 a (2.89)

Fatty acids composition of four diets Fatty acids (mg/g DW) 18:2(n-6) LLA 20:4(n-6) AA Total (n-6)hufa 18:3(n-3) LNA 20:5(n-3) EPA 22:6(n-3) DHA Total (n-3)hufa On-grown Artemia Artemia nauplii Moina Bloodworms 15.4 2.9 9.4 8.1 1.8 1.0 0.8 0.7 1.9 1.1 0.9 0.8 1.7 12.9 10.7 7.1 1.7 0.9 1.0 0.9 0.8 0.3 0.1 0.1 2.7 2.0 1.3 1.1 Total Fame 49.50 47.50 72.30 62.40 On-grown Artemia Deficient in LNA Highest in LLA AA, total (n- 6)HUFA, EPA & DHA AA might be essential to maturation and spawning of FW ornamental fish

Bioencapsulation of on-grown Artemia with DHA Continuous, non-selective feeding behavior - an ideal booster diets On-grown Artemia boosted with DHA show increases in EPA, DHA and (n-3)hhfa (n-3)hufa (mg/g DW 10 8 6 4 2 Control Boosted 0 LNA EPA DHA (n-3) HUFA

Bioencapsulation of on-grown Artemia with AA On-grown Artemia boosted with ascorbyl palmitate displayed significant increases in AA AA (ug/g WW) 100 80 60 40 20 0 Control ART ART 10% ART 20% b a b b a On-grown Artemia Guppy b Guppy fed boosted ongrown Artemia also resulted in concomitant increase in incorporated AA

Availability of on-grown Artemia would offer farmers and exporters a bigger alternative live feed for feeding larger ornamental fish and brooders More importantly, offer possibility of enhancing fish performance and quality through bioencapsulation

Conclusion The use of live feeds, such as FW rotifers, Artemia nauplii, decapsulated Artemia cysts and on-grown Artemia could be applied successfully in freshwater ornamental fish culture to improve fish performance Use of these live feeds are likely to have a positive impact to the ornamental fish industry

Application of bio-encapsulation Many possible applications, including boosting the ongrown Artemia with essential nutrient such as HUFAs to improve growth, survival & increase vigor pigments or color enhancer to obtain better coloration therapeutic drugs for disease treatment vitamin C or immuno-stimulants to enhance stress and disease resistance, and hormone to induce sex reversal, maturation & spawning

Discus: Parental feeding vs Artificial feeding Total length (mm) 12 10 8 6 4 2 0 AF30 AF20 PF20 a a a a a NM Rotifer (D7) Artemia (D14) Feeding phase AF30: Artificial feeding; 30 larvae/l; AF20: Artificial feeding; 20 larvae/l PF30 : Parental feeding; 30 larvae/l; NM: Not measured

Discus: Parental feeding vs Artificial feeding 100 Survival rate (%) 80 60 40 20 AF30 AF20 PF20 0 Rotifer (D2-7) Feeding phase Artemia (D8-14) AF30: Artificial feeding; 30 larvae/l; AF20: Artificial feeding; 20 larvae/l PF30 : Parental feeding; 30 larvae/l; NM: Not measured

Phase 1: Rotifer vs egg-yolk feeding, 30/l Phase 2: Both groups fed Artemia, 10/l Total length (mm) 14 12 10 8 6 4 2 Egg-yolk group Rotifer group b a a b Survival (%) 100 80 60 40 20 Egg-yolk group Rotifer group 0 Rotifer (D12) Artemia (D32) 0 Rotifer (D2-12) Artemia (D13-32) Feeding phase Feeding phase

Total length (mm) 12 10 8 6 4 2 0 AF30 AF20 PF20 a a NM a a a Rotifer (D7) Artemia (D14) Feeding phase