AN IMMUNE-MODULATOR ALGAL EXTRACT TO FIGHT AGAINST DISEASES THE EXAMPLE OF EMS IN L. VANNAMEI

Transcription

1 AN IMMUNE-MODULATOR ALGAL EXTRACT TO FIGHT AGAINST DISEASES THE EXAMPLE OF EMS IN L. VANNAMEI

2 DISEASE IN AQUACULTURE Shrimp industry losses Cost of shrimp disease in Asia (Shinn et al, 2016) Shrimp farming industry has been heavily touch by: 1. Bacteria Gram - : Acute Hepatopancreatic Necrosis Disease (AHPND/EMS) (V. parahaemolyticus) 2. Microsporidia : Enterocytozoon hepatopenaei (EHP) 3. Virus : White Spot Sydrome Virus (WSSV) as the most world wide spread Leading to tremendeous losses disreagarding the causative agent. 2

3 FAO-FDA approuve drugs 2010 & - O.L. Akinbowale 2006 Antimicrobial resistance in aquaculture DISEASE IN AQUACULTURE Antibiotics, a possibility but with majors constrains Declining Antibacterial approvals Bacterial resistance Florfenicol (Flo) Oxytetracycline dihydrate (Oxy) Sulfadimethoxine & ormetoprim (Sul) 20% of bacteria isolated from aquaculture setting in Australia show resistance to approved aquaculture drugs 3

4 ALGAE VARIETY BROWN SEAWEEDS RED SEAWEEDS GREEN SEAWEEDS VARIETY OF GENOME Variety of protein structure Variety of functions

5 ALGAE COMPOSITION Carbohydrates (20-70% dry weight) - mainly polysaccharides : - Alginates (brown algae) - Carraghenanes (red algae) - Ulvans (green algae) Proteins (1-35% dry weight) Lipids (0,5-4% dry weight) Minerals (10-30% dry weight) - Iron, Iodine, Copper, Potassium, Sodium, Calcium, - Magnesium, Sulfur, Phosphorous, Chlorine, Manganese - Many other trace elements. Vitamins - A, B1, B2, B6, B12, C, D3, E, K Pigments Phycoerythrin (antioxidant) Carotenoids 5

6 ALGAE ALTERNATIVE Biological activites Seaweeds (green, brown and red) sulfated polysaccharides have been identify to have: Anticoagulant and antithrombotic Anti-infectious Immune-modulating Antioxidant Antilipidemic Wound healing Branched sulfated hetero-polysaccharide Sulfate S S S S S S 6

7 OLMIX MSP s PROPERTIES Olmix Technology

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9 OLMIX MSP EXTRACTION Full traceability from collection to final products

10 nature review / cancert : IMMUNE SYSTEM Innate and Adaptative systems Unlikes terrestrial species, fish and shrimp moslty relies on the innate immune response 10

11 C. J. Secombes et al & C. Uribe 2011 & Liu 2009 IMMUNE SYSTEM 1st step: the pathogen recognition 1. Pathogen Associated Molecular Patterns (PAMPs): Lipopolysaccharide (LPS), Non Soluble Polysaccharides (beta 1.3-glucan, ) peptidoglycan, zymozan (yeast cell wall), other dead pathogen cell wall, 2. Pattern Recognition Proteins (PRPs) Are non-self-recognition factor that bind with PAMPs Lipopolysaccharide & glucans binding protein (LGBP, GBP), lectins 11

12 C. J. Secombes et al & C. Uribe 2011 & Liu 2009 IMMUNE SYSTEM 2nd step: innate immune response induced Toxic metabolites PO Opsonization TNFα, IL-1, IL-6 CCL2, CCL20 Specific protein/peptides MAC Phagocytosis Expression of messenger that promote the recruitment of immune cells Cytokines : TNFα, IL-1, IL-6, Chemokines: CCL2, CCL20 Innate response consist of: Phagocythosis (encapsulation of the pathogen by phagocythe Opsonisation (encapsulation to enhance phagocytosis) Membrane Attack Complex (MAC), it creates a hole in the pathogen s cell membrane) Toxic metabolites (ROs, NOs) & Phenol oxidase (PO) for shrimp 12

13 Alii Reza K. et al 2017 IMMUNE SYSTEM Pathogen proliferation risk Due to the time required to activate the all immune system, the pathogen may proliferate imparing growth and feed efficiancy 13

14 Yu Yuan 2016 IMMUNE MODULATION Olmix MSP immunity mode of action OLMIX MSPs are considered by the organism as PAMPs. Thus, they are recognized by the PRPs (TLR4/TLR2) Messenger molecules will be produced Macrophage and others immune cells recruitment will begin The natural immune defences will be modulate 14

15 IMMUNE MODULATION Olmix MSP immunity impact on messenger molecule production Berri et al Marine-sulfated polysaccharides extract of Ulva armoricana green algae exhibits an antimicrobial activity and stimulates cytokine expression by intestinal epithelial cells. J ApplPhycol. Vol 28, no1 & INRA Research study

16 IMMUNE MODULATION Olmix MSP immunity impact on messenger molecule production CYTOKINES AND CHIMIOKINES S EXPRESSION IN IPEC-1 EPITHELIAL CELLS Dose (mg/ml) Increased expression of several «Messenger molecules» of the immune system in differentiated IPEC-1 cells when they are stimulated with MSP at the concentration of 1mg/ml. (**: P<0.01) 16

17 Response IMMUNE SYSTEM Pathogen proliferation risk Innate response In case of pathogen infection, the all immune system will be ready to kill it before any proliferation Adaptative response Time after MSP s administration 17

18 IMMUNE MODULATION Conclusion MSP Immunity Innate Immunity Increase of the cytokine s* production *Production or recruitment vector of immune cells (i.e. -> chemokines, interferons, ) Stimulation of immune cells (i.e. -> macrophages, ) Prepare the organism to fight against pathogen Adaptive Immunity Migration and modulation of immune cells (i.e. -> Lymphocytes, ) From Berri et al, 2016 ; Chen et al, 2008; Liu et al.,

19 TRIALS 19

20 Searup EMS challenge trial L. vannamei Can Tho University, College of Aquaculture and Fisheries, Dept. Of Aquatic Pathology All PLs tested negative to WSSV, YHV, TSV, MBV and V. parahaemolyticus causing EMS by PCR method Shrimp PL5 raised in 4 (500L) tanks at 50pcs/L PL5 Treatment 1 Treatment 2 Treatment 3 Treatment 4 Challenge test PL 6 PL7 PL8 PL9 PL10 PL11 PL12 PL13 PL14 PL15 PL16 PL17 PL18 PL19 PL20 PL21 PL22 PL23 PL24 PL25 At stage PL25 shrimp were submitted to an immersion challenge with EMS causing bacteria (Vibrio parahaemolyticus).

21 Cummulative mortality (%) Searup EMS challenge trial L. vannamei Cumulative mortality post AHPND challenge Neg cont Pos cont 3d Searup 5d Searup 7d Searup Days post challenge

22 DOSAGE FARM USE Aquaculture use: Fish: Normal condition : 1-2ml/10kg live weight 2 times a week Under stress event/ farm troubles : 1-2ml/10kg live weight every day for 7 days Shrimp: Hatchery : 2ml/kg feed every feeding time Grow out : 2-4ml/kg feed 1 to 2 times a day MSP Immunity

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24 Reinforces natural defenses And compensates potential disbalances