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

B.S. Oceanography & M.S. Aquaculture

Technical Account Manager

Ridley Aquafeed - Australia

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

SUMMAR SUMMARY

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)

DEFINITI DEFINITION: ON: BIOFL BIOFLOCS OCS

Biofloc under microscope view, from left to right, 10x, 40x, 40x.

Biosecurity Strict environmental control Isolation from contamination sources Enhances prawn immune system Farm Efficiency Requires less land and water Reuse of feed wastes Allows higher densities, optimize number of crops, increases productivity Higher Quality Enhances animal health and produces stronger animals, increasing quality after harvest Environment Zero or limited water exchange Less effluent discharge Less waste and better waste management

DEFINITI DEFINITION: ON: BIOFL BIOFLOC OC TE TECHNOL CHNOLOGY OGY (BFT) (BFT) SY SYST STEM EM

HO HOW W IT W IT WORK ORKS? S?

TAN [ NH3 + NH4

+ ]

Feces Excretion Feed waste

NO2 NO3 (Non toxic)

Feeds

C org (C:N ratio)

Microbial Biomass

C inorg + Oxygen

Nitrifying Bacteria Physical Substrate

Microorganisms in the system (bioflocs) – two major roles: 1. Uptake of nitrogen compounds generating “in situ” microbial protein and 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

• Protein-rich
• Protein = 16% N
• N leaching and accumulation

Water Exchange & Nitrogen-rich Effluent discharges

NITR NITROGEN OGEN WAST WASTE E & & MICR MICROB OBIAL L PR PROTE TEIN IN

Discharge of effluents:

• Eutrophication of natural waters
• Ecological unbalance of the recipient environment
• Spread of diseases and contamination of wild

populations (permanent reservoirs) Waste Management

• Environmental regulations
• Market trend for organic and ”green”
• Increase efficiency of feeds

NITR NITROGEN OGEN WAST WASTE E & & MICR MICROB OBIAL L PR PROTE TEIN IN

BFT 70% less than conventional system

BFT

• Recycling nitrogen into bacterial protein
• Establishment of microbial food chain (protein-rich)
• Transfer of N into prawn biomass
• Maintenance of N in non-toxic levels
• Less generation of waste

NITR NITROGEN OGEN WAST WASTE E & & MICR MICROB OBIAL L PR PROTE TEIN IN

30% - 40% of prawn’s biomass is

• btained by biofloc consumption

in BFT system

(Burford et al., 2004; Cardona et al., 2015)

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
• Reduce feed demand
• Consequently increases efficiency

NITR NITROGEN OGEN WAST WASTE E & & MICR MICROB OBIAL L PR PROTE TEIN IN

Microorganisms – three major groups:

• 1. Heterotrophic bacteria
• 2. Chemo-autotrophic bacteria
• 3. Photo-autotrophic microalgae

2.3. 2.3. MICR MICROBI BIAL L COMM COMMUN UNITIES ITIES

• Assimilate Ammonia into protein
• Consume organic carbon
• Very fast cell duplication

HETEROTROPHIC BACTERIA

• Form the Bioflocs
• Proteobacteria, Bacteroidetes,

Bacillus spp.

Imhoff cone showing biofloc settled – Floc level will increase with the growth of heterotrophic bacteria Heterotrophic bacteria Bacillus spp.

MICR MICROB OBIA IAL L COMM COMMUNI NITIE TIES

Molasses as carbon source applications, followed by drop of TAN levels. Nitrogen assimilated as heterotrophic bacteria biomass. Da Silva et al., 2013.

• Assimilate Ammonia into protein
• Consume organic carbon
• Very fast cell duplication

HETEROTROPHIC BACTERIA

• Form the Bioflocs
• Proteobacteria, Bacteroidetes,

Bacillus spp.

MICR MICROB OBIA IAL L COMM COMMUNI NITIE TIES

Addition of Corg

• 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 (NO2)
• Nitrite Oxidizer Bacteria (NBO)
• Nitrobacter, Nitrococcus, Nitrospira
• Oxidize NO2 into nitrate (NO3) non toxic

CHEMO-AUTOTROPHIC BACTERIA

MICR MICROB OBIA IAL L COMM COMMUNI NITIE TIES

Nitrification process in biofloc system where nitrite is being oxidized into nitrate. Da Silva et al., 2013.

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

MICR MICROB OBIA IAL L COMM COMMUNI NITIE TIES

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/m3)

• 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/m3; 60% survival at 5000 PLs/m3)

• Basic initial cost: 15-25 USD/m2 *

Biofloc nursery for L. vannamei post larvae. Agua Blanca Seafood, Oaxaca, Mexico

* 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

and sources of contamination (birds, crabs, wind, etc)

Ecological interactions among the microbial community are more diverse in biofloc system when in outdoors conditions, what requires a stronger manipulation of the environment in order to set the functional roles of each 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

• In large industrial scale, mostly
• L. vannamei
• Basic initial cost: 7-10 USD/m2 *

* Cost based on HPED liner and aeration system. Values will vary according to regional availability and market price. Biofloc grow out ponds

• f L. vannamei. Above,

Agua Blanca Seafood, Oaxaca, Mexico; on the right, Southern Brazil.

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

• Basic initial cost: 15-25 USD/m2 *

* Cost based on HPED liner, aeration system and greenhouse structure. Values will vary according to regional availability and market price.

Indoors Biofloc prawn farming. On the right, Marvesta Shrimp Farms, Maryland, USA; below, Fazenda Cultivamar, Southern Brazil

APP APPLICA LICATI TION ONS

BROOD STOCK CULTURE

• Brood stock domestication
• Breeding Programs
• Avoid vertical contamination
• Generation of SPF lines
• Production of disease free PLs
• Culture under highest biosecurity levels
• Surveillance programs
• Artificial Insemination and individual validation

for new brood stock generation

Brood stock production facility operating in biofloc system in Saudi Arabia

• L. vannamei brood

stock reared in BFT in site previously affected by WSSV

• utbreak

MAN MANAGEMENT GEMENT

Specialized management

• Qualified personnel

Dissolved Oxygen / Aeration

• High O2 demand: respiration and microbial processes
• Suspended solids

pH and Alkalinity

• Acidification; nitrification; buffer effect

Different kinds of aeration to keep biofloc in suspension and

• xygenate the water: above, paddle-wheels that provide

horizontal circulation; and on the left, microporous hoses that generate vertical circulation cells

MAN MANAGEMENT GEMENT

Solids

• Excess: DO issues, gill fouling, sludge
• Scarcity: poor microbial process and

control of nitrogenous wastes

Imhoff cone showing scarcity (left) and excess (right) of floc. Clarifiers for solid control in BFT system in Saudi Arabia Sludge accumulated by solid excess

MAN MANAGEMENT GEMENT

Algae community

• Outdoors and greenhouses
• Balanced microbial community
• Filamentous species and blue-green algae

Filamentous algae bloomed in biofloc prawn culture Blue-green algae Nodularia spp. contamination of biofloc pond (right) in southern Brazil.

MAN MANAGEMENT GEMENT

Feeding

• Adjusted feed rates; prawn and

microbial performance

• High quality feed = high quality floc

Nitrogen

• TAN / NO2 / NO3
• Constant and accurate monitoring
• Calculation of carbon applications
• Affects feeding management

Water quality monitoring by spectrophotometry 32% protein prawn feed produced in Brazil

BIOSE BIOSECUR CURITY TY

BIOFLOC & BIOSECURITY Physical Barrier

• Minimum water exchange
• Rigorous water treatment
• Indoors culture
• Higher environmental control

Biological Barrier

• SPF brood stock and larvae
• Beneficial bacteria (probiotic effect)
• Ecological competition
• Enhancement of immune system

BIOSE BIOSECUR CURITY TY

• Biofloc has bioactive compounds that contribute for a

healthy status of cultured prawns (Ju et al., 2008b)

• Expressions of certain hemocytes enzymes related to

immune system is enhanced in biofloc reared L. vannamei (Jang et al., 2011)

• Bioflocs have positive effect in the immune response of
• L. vannamei leading to higher resistance against IMNV

challenge (Ekasari et al., 2014)

• Immune system and antioxidants enhanced in L.

vannamei juveniles reared in biofloc (Xu & Pan, 2013)

• Prawns show resistance to Vibrio spp when reared in

BFT (Liu et al., 2017)

(Ju et al., 2008b)

BIOSE BIOSECUR CURITY TY

Biofloc: higher degree of biosecurity

• Limited water exchange
• Higher environmental control
• Physical barrier against pathogens

(indoors)

• Biological barrier against pathogens
• Enhances prawn immune system

Biosecurity Protocols

• Hygiene and sanitation
• Training of personnel
• Disinfection of facilities, materials, vehicles, staff
• Control and validation of raw materials
• Biosecurity zones and controlled movement
• Surveillance program

SPF & Tolerant lines

• Vertical contamination
• SPF larvae
• Head start for producer
• Brood stock domestication importance
• Tolerance against diseases

Water Treatment

• Need of sterile water
• Filter bags
• Sand, cartridge and carbon filters
• UV system
• Ozone system

EXA EXAMP MPLES LES – LA LAGUN GUNA, A, SC SC - SOUT SOUTHERN HERN BR BRAZ AZIL IL

• L. vannamei farming completely devastated by

WSSV outbreak in early 2000’s

• Contamination of natural water bodies and all

farms in the region

• Biofloc pilot installed in the epicentre of
• utbreak
• Basic remodelling of the farm: smaller,

lined ponds, bird nets, crab fences, basic biosecurity protocol, stocking of SPF PLs

Remodelling of WSSV contaminated ponds into biofloc operational units; implementation of basic biosecurity measures – bird nets, crab fences, vehicle disinfection area. Laguna, SC - Brazil

EXA EXAMP MPLES LES – LA LAGUN GUNA, A, SC SC - SOUT SOUTHERN HERN BR BRAZ AZIL IL

Crops Stock density (pcs/m2) FCR Survival Yield (kg ha-1) WSSV

• ccurrence

1 100 1.32 63% 6.36 NEGATIVE 2 118 1.25 75% 7.97 NEGATIVE 3 100 1.15 75% 6.75 NEGATIVE

Results of pilot crops in biofloc system operated in WSSV contaminated area in Laguna, SC – Southern Brazil. Poersch et al., 2013 Panorama da Aquicultura Magazine

Panoramic view of the farm after remodelling to operate in Biofloc system.

• 3

successful crops regularly monitored through PCR and histopathological analysis – all WSSV negative

EXA EXAMP MPLES LES – SA SAUDI UDI ARABI ARABIA

• First crop 2015: 15.000 t
• Second crop 2016: 17.500 t
• Target for 2017: 30.000 t
• Surveillance program: all WSSV negative
• P.

indicus farming completely devastated by WSSV

• utbreak in early 2010’s - Vertical contamination
• Introduction of SPF Tolerant L. vannamei
• Implementation of biosecurity protocols
• Biofloc system for the production of brood stock and

implementation of breeding program

• L. vannamei prawn farms in Saudi Arabia – Rigorous biosecurity strategy and

biofloc-reared brood stock guarantee success in conventional system grow

• ut.
• L. vannamei SPF

brood stock reared in BFT system in Saudi Arabia

CONCL CONCLUSI USION ON

S W T O

• Higher Biosecurity
• Higher Productivity
• Less use of water and land
• Less environmental impact
• Higher investment
• Remodelling of existing units
• Require qualified

management

• Operate in areas affected

by WSSV outbreak

• Operate in seasonal

periods and continental areas where conventional system cannot

• Needs to be a part of a

rigorous biosecurity strategy in order to thrive

• Risk of system crash if

not well managed

CONCL CONCLUSI USION ON

Disease outbreaks, particularly WSSS, typically cause major constraints to prawn

• aquaculture. Industries in Australia and around the world must adapt their practices to

minimize and/or prevent the impact of these outbreaks. New technologies, such as Biofloc and RAS, are available to assist in this challenge. Although the initial investment is higher than other systems, best practices worldwide have shown that BFT is suitable for industrial prawn aquaculture. It increases farm efficiency, production, and enhances the immune-system of the prawns and on-farm

• biosecurity. Such characteristics make BTF a powerful tool to produce in WSSV affect

areas, although it needs to be part of a much larger biosecurity strategy. Transfer of technology, system remodeling, adaptations, and proper management must be made to expand the scope of the commercial production in BTF to other species, such as P. monodon. Transitioning to BTF will help improve prawn farming, which will lead the aquaculture industry toward a higher level of sustainability and animal health.

Gabriel B. Santos Technical Manager – Shrimp Breeding Program B.S. Oceanography & M.S. Aquaculture e-mail: oc.gabrielsantos@gmail.com