BIOF BIOFLOC OC AS AS A A BIOSE BIOSECUR CURITY ITY TOOL - PowerPoint PPT Presentation

BIOF BIOFLOC OC AS AS A A BIOSE BIOSECUR CURITY ITY TOOL OOL AGAINST GAINST WSSV WSSV Gabriel B. Santos Marcell B. de Carvalho Technical Manager of Shrimp Breeding Program Technical Account Manager B.S. Oceanography & M.S. Aquaculture Ridley Aquafeed - Australia

SUMMARY SUMMAR  DEFINITION  OVERVIEW OF THE SYSTEM  How it works  Nitrogen waste and Microbial Protein  Microbial Communities  APPLICATION & MANAGEMENT  BIOSECURITY  EXAMPLES  CONCLUSION

DEFINITI DEFINITION: ON: BIOFL BIOFLOCS OCS Aggregates formed by a complex interaction between particulate organic matter and a large range of microorganisms, such as bacteria and phytoplankton, and grazers, such as rotifers, ciliates and flagellates protozoa and copepods. (Avnimelech, 2007; Ray et al. , 2010; Emerenciano et al. , 2013) Biofloc under microscope view, from left to right, 10x, 40x, 40x.

DEFINITI DEFINITION: ON: BIOFL BIOFLOC OC TE TECHNOL CHNOLOGY OGY (BFT) (BFT) SY SYST STEM EM Biosecurity Higher Quality Strict environmental control Enhances animal health and produces Isolation from contamination sources stronger animals, increasing quality Enhances prawn immune system after harvest Farm Efficiency Environment Requires less land and water Zero or limited water exchange Reuse of feed wastes Less effluent discharge Allows higher densities, optimize Less waste and better waste number of crops, increases productivity management

HO HOW W IT W IT WORK ORKS? S? C org C inorg + Feeds (C:N ratio) Oxygen NO 3 (Non toxic) Physical Substrate NO 2 Microbial Nitrifying Biomass Bacteria TAN + ] Feces [ NH 3 + NH 4 Excretion Feed waste

NITROGEN NITR OGEN WAST WASTE E & & MICR MICROB OBIAL L PR PROTE TEIN IN Microorganisms in the system (bioflocs) – two major roles: Uptake of nitrogen compounds generating “in situ” microbial protein and 1. maintaining water quality; and 2. Increasing culture feasibility by reducing FCR and a decrease of feed costs by reducing protein demand

NITR NITROGEN OGEN WAST WASTE E & & MICR MICROB OBIAL L PR PROTE TEIN IN Feeds Water Exchange • Protein-rich & • Protein = 16% N Nitrogen-rich Effluent discharges • N leaching and accumulation

NITR NITROGEN OGEN WAST WASTE E & & MICR MICROB OBIAL L PR PROTE TEIN IN Discharge of effluents: Waste Management • • Eutrophication of natural waters Environmental regulations • Market trend for organic and ”green” • Ecological unbalance of the recipient environment • • Spread of diseases and contamination of wild Increase efficiency of feeds populations (permanent reservoirs)

NITR NITROGEN OGEN WAST WASTE E & & MICR MICROB OBIAL L PR PROTE TEIN IN BFT • Recycling nitrogen into bacterial protein • Establishment of microbial food chain (protein-rich) 70% less than • Transfer of N into prawn biomass conventional system • Maintenance of N in non-toxic levels • Less generation of waste BFT

NITROGEN NITR OGEN WAST WASTE E & & MICR MICROB OBIAL L PR PROTE TEIN IN Crude Protein Reference 43% McIntosh et al., 2000 12 - 42% Soares et al., 2004 26 - 41.9% Ju et al., 2008 31% Tacon et al., 2010 38.8 - 40.5% Kuhn et al., 2010 28 - 43% Maicá et al., 2012 • Reduce FCR 30% - 40% of prawn’s biomass is obtained by biofloc consumption • Reduce feed demand in BFT system • Consequently increases efficiency (Burford et al., 2004; Cardona et al., 2015)

2.3. 2.3. MICR MICROBI BIAL L COMM COMMUN UNITIES ITIES Microorganisms – three major groups: 1. Heterotrophic bacteria 2. Chemo-autotrophic bacteria 3. Photo-autotrophic microalgae

MICROB MICR OBIA IAL L COMM COMMUNI NITIE TIES Heterotrophic HETEROTROPHIC BACTERIA bacteria Bacillus spp . • Assimilate Ammonia into protein • Consume organic carbon • Very fast cell duplication • Form the Bioflocs • Proteobacteria, Bacteroidetes, Bacillus spp. Imhoff cone showing biofloc settled – Floc level will increase with the growth of heterotrophic bacteria

MICROB MICR OBIA IAL L COMM COMMUNI NITIE TIES HETEROTROPHIC BACTERIA Addition of C org • Assimilate Ammonia into protein • Consume organic carbon • Very fast cell duplication • Form the Bioflocs • Proteobacteria, Bacteroidetes, Bacillus spp. Molasses as carbon source applications, followed by drop of TAN levels. Nitrogen assimilated as heterotrophic bacteria biomass. Da Silva et al., 2013.

MICROB MICR OBIA IAL L COMM COMMUNI NITIE TIES CHEMO-AUTOTROPHIC BACTERIA • Nitrifying Bacteria • Late establishment in the system • Require to be attached for effective nitrification • Consume inorganic carbon (alkalinity) • Probiotics and/or Inoculum • Ammonia Oxidizer Bacteria (AOB) • Nitrosomonas, Nitrosococcus, Nitrosospira • Oxidize ammonia into nitrite (NO 2 ) • Nitrite Oxidizer Bacteria (NBO) • Nitrobacter, Nitrococcus, Nitrospira • Oxidize NO 2 into nitrate (NO 3 ) non toxic Nitrification process in biofloc system where nitrite is being oxidized into nitrate. Da Silva et al., 2013.

MICR MICROB OBIA IAL L COMM COMMUNI NITIE TIES PHOTO-AUTOTROPHIC MICROALGAE • Light penetration • Outdoor ponds and greenhouse enclosed systems • Daily fluctuations • Diatoms • Filamentous algae and blue-green algae • Management to balance communities

MICR MICROB OBIA IAL L COMM COMMUNI NITIE TIES Immature System • Heterotrophic pathway 100% of N recycling • Carbon addition necessary • Increase in bioflocs (surface area) • Protein source and immune system Mature System • Nitrification process established • Chemo-autotrophic pathway – 65% of N recycling • Heterotrophic pathway – 35% of N recycling • Carbon provided by feeds (organic) and alkalinity (inorganic) Outdoors and abundant light conditions • Microalgae • Can be beneficial if well managed • Synergic balance among communities

APP APPLICA LICATI TION ONS NURSERY PHASE • High-biosecurity facilities to grow post- larvae (0.3 – 3.0g) • Very high stocking densities and biomass (500 – 10.000 PL/m 3 ) • Management of nitrogenous wastes • Improves immune system • Stock grow out with more resistant juveniles High density nurseries provide safe environment for the most sensible life stage after hatchery. When stocked in the farms, prawns are stronger and usually show compensatory growth. In the image, a greenhouse enclosed nursey operating in BFT system in southern Brazil.

APP APPLICA LICATI TION ONS NURSERY PHASE • USA, Mexico, Central America, Ecuador, Brazil, Saudi Arabia, Southeast Asia – mostly for L. vannamei • Also successfully applied for F. paulensis, F. brasiliensis, F. setiferus and P. monodon • P. monodon (88% survival at 1000 PLs/m 3 ; 60% survival at 5000 PLs/m 3 ) Biofloc nursery for L. vannamei post larvae. Agua Blanca Seafood, Oaxaca, • Mexico Basic initial cost: 15-25 USD/m 2 * * Cost based on HPED liner, aeration system and greenhouse structure. Values will vary according to regional availability and market price.

APP APPLICA LICATI TION ONS OUTDOORS GROW OUT • Lined, smaller ponds • Higher stocking density • Higher aeration power • Algae presence • Biofloc Inoculum • Biofloc system = higher animal health & biosecurity • Susceptible to environmental conditions Ecological interactions among the microbial community are more diverse in and sources of contamination (birds, biofloc system when in outdoors conditions, what requires a stronger manipulation of the environment in order to set the functional roles of each crabs, wind, etc) community in synergy. In the image, outdoor grow out pond operating in BFT system in southern Brazil

APP APPLICA LICATI TION ONS OUTDOORS GROW OUT • Central and South America, Southeast Asia Biofloc grow out ponds of L. vannamei . Above, • In large industrial scale, mostly Agua Blanca Seafood, Oaxaca, Mexico; on the L. vannamei right, Southern Brazil. • Basic initial cost: 7-10 USD/m 2 * * Cost based on HPED liner and aeration system. Values will vary according to regional availability and market price.

APP APPLICA LICATI TION ONS INDOORS GROW OUT • Higher stocking densities = less land and smaller production units • Higher initial investment • Automatization • Higher environmental control • Barriers against contamination sources = higher biosecurity • Allows production in land and in seasonal periods of low temperature • Operation in sites previously affected by WSSV

APP APPLICA LICATI TION ONS INDOORS GROW OUT • USA, Central and South America, Saudi Arabia, Korea, China Indoors Biofloc prawn farming. • Basic initial cost: 15-25 USD/m 2 * On the right, Marvesta Shrimp Farms, Maryland, USA; below, Fazenda Cultivamar, Southern Brazil * Cost based on HPED liner, aeration system and greenhouse structure. Values will vary according to regional availability and market price.