Seventh Framework Programme

Research for SMEs 2 years: 2011-2013 Budget: 1,112.388.65 €

SEMs RTDs

North Bay Shellfish Ltd

-

2.000

4.000

6.000

8.000

10.000

12.000

14.000

-

2.000.000

4.000.000

6.000.000

8.000.000

10.000.000

12.000.000

14.000.000

16.000.000

1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010

Valu

e (m

illio

n U

SD)

Prod

uctio

n (t

)

Aquaculture

Capture

Aquaculture, value

Global Bivalve Production

1. TOXIC EPISODES

In 2010 the costs of HABs in Galicia were estimated in 40,000,000 € (Research project purgademar) http://www.purgademar.com

In Europe, estimation of the economic impact of HABs on farmed shellfish was $ 754 Million from 2000 to 2009 which was an annual value of $ 75.4 Million (FAO)

2. MICROBIAL AND PARASITIC CONTAMINATION

Bacteria: E coli, Salmonella spp, L monocytogenes, Vibrio sp Virus: HAV , MNV (winter vomiting bug) Parasites: Bonamia ostreae

List of Work packages (WPs)

WP1: Investigate the use of detoxifying bacteria delivered by microencapsulation to sites of toxins in shellfish WP2: Investigate the use of administered microcapsules as an aid to effective and efficient depuration of pathogens WP3: Investigate the use of immunostimulants or delivery of direct anti-Bonamia agents WP4: Demonstration of results of detoxification/depuration at commercial depuration plants WP5: Evaluation. Assessment of IPR, Dissemination activities and Training WP6: Project Management

WP1: Investigate the use of detoxifying bacteria delivered by microencapsulation to sites of toxins in shellfish

Objectives: 1. To characterize and identify toxin

degrading bacteria

2. To determine properties that may significantly influence degradation of algal toxins by bacteria

3. To develop a “probiotic” diet of toxin degrading bacteria and a microencapsulation delivery system for shellfish consumption with targeted release in the DG to degrade toxins

Objectives

1. To design protocols using fluorescent microbeads to approve the suitability of shellfish purification systems

2. To define a panel of bacteria with potential activity against pathogens

3. To determine the best method for microencapsulation of selected bacteria

4. To investigate viability of applying microencapsulated diets to increase the depuration process of shellfish from pathogens

WP2: Investigate the use of administered micro-capsules as an aid to effective and efficient depuration of pathogens

WP3: Investigate the use of immunostimulants or delivery of direct anti-Bonamia agents

Objectives

1. Determine the size and types of microencapsulated

beads to use in oysters tissues

2. Optimize the delivery of microencapsulated beads

or immunostimulats to oysters infected with B ostreae

3. To determine if those enhance the immune

response of the infected oyster

4. To prove that there is a reduction in the prevalence

or intensity of infection

WP 4: Demonstration of results of detoxification/depuration at commercial depuration plants

Objectives

• To determine the potential of probiotic diets to detoxify shellfish in a commercial setting.

• To design “alternative” protocols using fluorescent microbeads (as a proxy for bacteria/virus) for shellfish depuration system approval.

• To determine the increase of the efficiency of shellfish depuration of bacteria and viruses using microencapsulated probiotics.

WP5: Evaluation. Assessment of IPR, Dissemination activities and Training WP6: Project Management

Objectives

1. To evaluate the results of the project

2. Dissemination and Exploitation of the results

3. To train key personnel in the SMEs and others

4. To manage the project: meetings, tasks, financial administration

Main Common Tasks WP1 and WP2 1. Isolation and characterization of toxin or pathogens degrading bacteria focusing on degradation of: ASP and DSP toxins Bacteria and Virus 2. Define the best method for construction of microcapsules-containing bacteria 3. Delivery of microencapsulated “probiotic” diets in the DG of shellfish to improve depuration and toxin degradation

MAIN ACHIEVMENTS OF THE BEADS PROJECT A protocol using fluorescent microbeads was designed to substitute the microbiological challenge test Almost 1000 bacteria isolated from shellfish and screened for activity against pathogens or toxins Some isolates, as bacteriocin-like substances show activity against L monocytogenes or E coli Compounds as zymosan microencapsulated present an inmunoestimulant effect on oysters

MAIN ACHIEVMENTS OF THE BEADS PROJECT

Some bacteria have potential to degrade toxins: OA and DA A method for microencapsulation and assessment of incorporation of bacteria or isolates into the DT of shellfish was developed Alginate capsules are a useful tool to transport probiotic bacteria and release them into de DG During passage through the intestine, the alginate beads are broken, so bacteria are released into the DT

20x

MAIN ACHIEVMENTS OF THE BEADS PROJECT

Alginate capsules can be used for fluorescent beads transport to the DG. These can be

monitored during their incorporation and later release in the DG of mussels and oysters and

then used in challenge tests.

Challenge test

Alginate capsules containing non degradable fluorescent dye microbeads in faeces of mussels.

MAIN ACHIEVMENTS OF THE BEADS PROJECT

Selection and microencapsulation of suitable probiotic bacteria or extracts

1. Encapsulation of 3M.21 In vitro activity:

2. Encapsulation of 3M18 Anti MNV-1 in vitro

3. Encapsulation of 4O11 Anti MNV-1 in vitro

Anti Listeria

Anti E. faecalis Anti Hepatitis A Anti MNV-1 activity

MAIN ACHIEVMENTS OF THE BEADS PROJECT

Delivery of beads containing probiotics to the target shellfish

• Alginate capsules transport probiotic bacteria and release them into DGs since they have been observed by light microscopy as intact in the DG of shellfish during feeding period. • Broken alginate beads were found in faeces, so during passage through the intestine bacteria embedded by alginate capsules are released.

•Probiotic diets embedded by alginate capsules can be released to improve depuration

Alginate capsules, empty and with a probiotic bacteria.

WP4. Demonstration of results of detoxification/depuration

at commercial depuration plants

Objectives

• To determine the potential of probiotic diets to detoxify shellfish in a commercial setting.

• To design “alternative” protocols using fluorescent microbeads (as a proxy for bacteria/virus) for shellfish depuration system approval.

• To determine the increase of the efficiency of shellfish depuration of bacteria and viruses using microencapsulated probiotics.

Months 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

WP4. Demonstration of results of detoxification/depuration at commercial depuration plants

Degradation of microalgal toxins

Depuration of bacteria/viruses

Challenge test

Assessment of: Toxin concentrations Microbeads clearance rates Bacterial and viral clearance rates

Microbeads Demonstration at commercial depuration plants

Report on demonstration of the challenge test

WP4. Demonstration of results of detoxification/depuration

at commercial depuration plants

Report on demonstration of the challenge test

WP4. Demonstration of results of detoxification/depuration

at commercial depuration plants

Conclusion: Microencapsulated probiotic bacteria can reach and colonize the DG of shellfish and then improve shellfish depuration Outcomes of this project belong to the industries of the consortium. They decided to put together some 'heads of terms' which is the basis for a joint venture or commercial collaboration in order to exploit the results reached in the project

A B C

10x 10x 10x

North Bay Shellfish Ltd

Thanks for your atention