Diet & Microbial Interactions ti In Palinurid Lobster Larvae Presented by: Mike Hall
Marine Lobsters Nephropoidea Palinuridea Scyllaridae Various Jasus spp. Panulirus cygnus Tropical Panulirus spp. Homarus americanus Panulirus argus Panulirus argus Nehrops norvegicus
Lobster Biodiversity Species: Wild Fisheries and Aquaculture Potential Group Species Wild Fishery Major Minor/ None Aquaculture Potential Seafood Aquarium Family Palinuroidea 47 31 16 +++ ++ Family Scyllaridae Subfamily Arctidinae 16 - Some ++ + Subfamily Ibacinae Subfamily Scyllarinae Subfamily Theninae 13 +40 5 - - - Some - 5 + - +++ + ++ + Family Synaxidae 3 - - - +++ Family Nephropoidea Subfamily Neophoberinae Subfamily Thymopinae 2 20 - - - - - + - - Subfamily Nephropinae 26 3 Some +(++) + Family Thaumastochelidae 3 - - - - Family Enoplometopoidae +11 - - - +++ TOTAL +176 34 Some Several Some
Seafood Groups as Production Units Price per kg per whole animal Fish Crustacean Molluscs Cephalopods 70 60 50 40 30 US $/kg 20 10 European Lobster American Lobster Mantis shrimp Whiteleg shrimp Rose shrimp Black Tiger Shrimp c/w Octopus Sepia Squid Carp Nile Perch Trout Salmon Eel Misc Mackerel Sprat Swordfish Skipjack Yellowfin Bluefin Sole Turbot Cod 0 Clam Scallop Oyster Mussel Spiny Lo lob obster Source: Globefish, European Price Report 2008
Australian Rock Lobster Biodiversity Potential Aquaculture Candidates (9 species) P. polyphagus P. homarus P. versicolor P. ornatus $22 mill. P. penicillatus P. longipes P. cygnus $281 mill. J. edwardsii $152 mill. J. verreauxi $5.4 mill.
Model of Aquaculture Propagation System Closed life cycle Wild stock Broodstock Maturation ti Harvesting Processing Marketing Closed-life Cycle (Farming) Closed-cycle l Spawning Farming Independent of wild stock Selective breeding Biosecurity Domestication Grow- out Hatching Larval Rearing
Closed-life cycle Propagation System Wild stock (Captured Rearing) Broodstock Maturation Spawning Harvesting Processing Marketing Growout? Larval Rearing
Larval Periods of Selected Crustacea Panulirus ornatus P. cygnus Jasus verreauxi J. edwardsii Homarus Crabs Cherax spp. Penaeus spp. Aquaculture Species 0 4 8 12 16 20 24 Time (months)
Palinuridae Phyllosoma Larval Rearing Larval Cycle Completed in > 9 species Species Panulirus species Reference Panulirus argus Moe (1991), Goldstein et al. (2008) Panulirus elephas Mercer et al. (1997), Kittaka et al. (1988), Kittaka et al. (2000) Panulirus homarus Radhakrishnan and Vijayakumaran (1993) Panulirus japonicus Oshima (1936), Kittaka and Kimura (1989), Yamakawa et al. (1989), Sekine et al. (2000) Panulirus interruptus Johnson (1956), Dexter (1972) Panulirus lalandii Kittaka (1988) Panulirus longipes Matsuda and Yamakawa (2000) Panulirus ornatus MGKailis (2006), AIMS (2007) Panulirus pencillatus Matsuda et al. (2006) Panulirus polyphagus Saisho (1966), Sin (1967) Jasus edwardsii Jasus species Kittaka et al. (1988), Booth (1996), Illingworth et al. (1997), Tong (1997), Moss (2000), TAFI (2005) Jasus verreauxi Kiattaka et al. (1997), TAFI (2006) Thenus orientalis Mikami and Greenwood (1997) Scyllarid species
Smith, G., Salmon, M., Kenway, M. and Hall, M. (2009) Description of the larval morphology of captive reared Panulirus ornatus spiny lobsters, benchmarked against wild caught specimens. Aquaculture 295:76-88.
Change in Larval Size through Development 11 Stages
Model of Larval Survival Continuous larval attrition and mass mortality events 100,000 90,000 80,000 Mortality/month 10% 6 month larval phase Progen ny 70,000 60,000000 50,000 40,000 30,000 20,000 10,000 0 20% 30% 40% 50% 0 1 2 3 4 5 6 Months 10% Survival
Conceptual Model Interactions between Nutrition Health - Growth 6 Metabolomics Healthomics Nutrition Health Growth Nutrigenomics Microbiomics
Nutrition Disease - Growth 6 Nutrition Health Growth Growth
Poor Nutrition Health - Growth 6 Nutrition Health Growth Growth
Poor Nutrition Disease - Growth 6 Nutrition Health Growth Morbidity Death Growth
Optimising the Triad Interactions between Nutrition Disease - Growth 6 Metabolomics Healthomics Nutrition Health Growth Nutrigenomics Microbiomics
Microbial Inputs Microbial Biosecurity Primary Input 1 Seawater 2 Added daily 2. Feed Artemia 60:1 x 100 s days Feed Hatchery Tank 3 Biofilm Planktonic bacteria 4 Phyllosomas Continuous Added once 3. Biofilm Complex community 4. Phyllosomas 90% of cells are bacteria Random Other vectors Aerosols, staff Discharge
Microbial Sources and Sinks in Hatchery Four Compartments 1. Seawater Column 2. Larval Feeds 3. Biofilm 4. Phyllosoma External Internal
Biofilm Complex Bacterial Ecosystem Diving through the biofilm Surface to Tank (2 µm steps)
Interactions between Nutrition Disease - Growth Larval finfish consume feed whole - Feed size related to mouth gape Source: Artemia World
Interactions between Nutrition Disease - Growth Phyllosomas consume feed piecemeal Stage 1 Stage 7
Health - External Fouling Mandibles Paragnaths 1st Maxilla Endites Basal Coxal
Changes in Mouthpart through Development Stages 1 to 11 Stage 1 Stage 11
Larval phyllosomas consume feed piecemeal Feeding video artemia
External Bacterial Fouling of Phyllosoma Mouthparts Day 1 Day 3 Post-molt Post-molt
External Bacterial Fouling of Phyllosoma Mouthparts Day 11 Post-molt Day 13 Post-molt FISH probe - Thiothrix Immediate Post-molt
Fouling (Thiothrix) of Phyllosoma Mouthparts Stage 1 to 2 ree of Fo ouling Deg 10 9 8 7 6 5 4 3 2 1 0 S1 to S2 Typical Moult Cycle 7days 0 1 2 3 4 5 6 7 8 9 10 11 12 13 Day since Moult
Fouling (Thiothrix) of Phyllosoma Mouthparts Stage 3 to 4 compared to Stage 1 to 2 Deg gree of Fouling 10 9 8 7 6 5 4 3 2 1 0 S1 to S2 S3 to S4 Typical Moult Cycle 7 days 0 1 2 3 4 5 6 7 8 9 10 11 12 13 Day since Moult
Interactions between Nutrition Health - Growth Fouled Hatchery compared to Unfouled 12.0 10.00 FOULED WILD Body Length (m mm) 8.0 6.0 4.0 2.0 0.0 0 1 2 3 4 5 6 7 8 9 10 11 Developmental Stage
Interactions between Nutrition Health - Growth Unfouled Hatchery compared to Wild 35.0 WILD HATCHERY 30.00 Body Length (m mm) 25.0 20.0 15.0 10.0 5.0 0.0 0 1 2 3 4 5 6 7 8 9 10 11 Developmental l Stage
Interactions between Nutrition Health Growth Internal Infection
Interactions between Nutrition Health Growth Ingestion and Digestion
Interactions between Nutrition Health Growth Internal Infection Wild-caught larva with intact hepatopancreas Hatchery larva with white-gut syndrome, infected by Vibrio spp.
Phyllosoma gut infection Real-time video
Health Internal Infections Survival of P. ornatus phyllosomas (S5) challenged with Vibrio sp. (DY05) Surv vival (%) 100 90 80 70 60 50 40 30 20 10 0 0 1 2 3 4 5 6 7 8 9 10 Time (Days) Control DY05
Phyllosoma gut Internal bacterial colonization Healthy Moribund Histopathology of hepatopancreas (H&E) Healthy: caeca open and clear Moribound: caeca are blocked and disrupted
Fluorescent in situ hybridisation (FISH) Bacterial localisation Healthy Moribund Probes: Red = general bacteria, Purple = Vibrio
Bacterial Diversity in Hatchery Phyllosoma Clone Library Analysis Only single representatives: Rare Operational Taxonomic Units (OTUs) Thiothrix Vibrio Bacteroidetes Photobacterium Sulfitobacter Thiothrix Vibrio Bacteroidetes Photobacterium Sulfitobacter Episilonbacter Roseobacter Exiguobacterium Psychrobacter Single OTUs
Interactions between Nutrition Health - Growth Oceanic Hunting Probiotics Tank Health Natural Microbial community Health Probiotic Microbial community
Larval Ecology of P.ornatus in NE Australia Phyllosoma entrained in the Coral Sea Gyre
Bacterial Diversity in Wild Phyllosoma Clone Library Analysis Psychrobacter Only single representatives: Rare Operational Taxonomic Units (OTUs) Sulfitobacter Episilonbacter il Thiothrix Vibrio Bacteroidetes Photobacterium Sulfitobacter Episilonbacter Roseobacter Exiguobacterium Psychrobacter Single OTUs Exiguobacterium Roseobacter
Bacterial Community Composition Hatchery vs Wild Hatchery Wild External Internal Internal Thiothrix Vibrio Bacteroidetes Photobacterium Sulfitobacter Epsilonbacteria i Roseobacter Exiguobacterium Psychrobacter Single OTUs 60 50 40 30 20 10 10 20 30 40 50 60 % Bacterial Clone Library Representation
The search for Probiotics Well diffusion assay agar infused with strain DY05 Homogenise Replica plating Sample natural environment 16S rdna sequencing Identification + AIMS culture collection - - - + + - Well Diffusion Assay Target organism: Vibrio sp. DY05
Sourcing Probiotic Strains 18% (92/508) of isolates inhibited growth of known P. ornatus pathogens (Vibrio strains DY05 & D40) in well-diffusion assays 4% 2% 1% 2% Zooplankton 39% 52% Wild Phyllosomas Hatchery Phyllosomas AIMS Hatchery Seawater Other
Identifying the Probiotic Candidates 16S rdna sequencing of 60 candidate probionts Psychrobater Roseobacter 13% Bacillus 3% 2% 2% 2%2% Vibrio Pseudoaltermonas 76% Pseudoaltermonas Vibrio Roseobacter Psychrobacter Bacillus Ferrimonas Acinetobacter
Current and future direction Interactions ti between Nutrition Health - Growth Elucidation of pathogen-probiont p interactions in vivo Vibrio (DY05) Infection GFP labelled Vibrio (DY05) At Artemia gut
Closed life cycle Aquaculture Panulirus us ornatus Wild stock Broodstock Maturation ti Spawning Hatching Harvesting Processing Marketing Grow- out Larval Rearing
Wild stock (Captured Rearing) Closed life cycle Aquaculture Panulirus ornatus Broodstock Maturation Closed-life Cycle (Farming) Spawning Harvesting Processing Marketing Growout Larval Rearing
Staff Investment Area Water Quality Broodstock Facility Larval Rearing Facility Live Feed Facility Health (Microbiology) Research Diet (Larval Feeds) Research Cruises Natural History, Prey, Probiotics, Diet Associated Larval Research Staff Matt Kenway Jane Gioffre Michael Clarkson Katie Holroyd Matt Salmon Grant Milton Justin Hochen Greg Smith Lone Høj Mike Hall Greg Smith Mike Hall Above staff plus volunteers 6 PhDs, 1 MSc Evan Goulden Ana Cano Gomez