Sécuritésanitaire en élevage avicole en climat chaud : quelles stratégies de prévention? Alain Réocreux September 2014 OlmixFrance

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1 Sécuritésanitaire en élevage avicole en climat chaud : quelles stratégies de prévention? Alain Réocreux September 2014 OlmixFrance

2 Nanotechnologyapplications in Animal Feeds Eco Concept M T.X+ Toxin Binder M Feed Natural Growth Promoter

3 UMR CNRS 7016 LMPC Amadeite, the origins to his industrial applications a b c Discovered in 2005 by Olmix in France

4 UMR CNRS 7016 LMPC Collaboration between Olmix, LPMC and CEVA UMR CNRS 7016 LMPC

5 Amadeite, Amadeite, the origins to his its industrial applications Why using Montmorillonite? oxygen silicon a b c hydroxyl or oxygen aluminium and magnesium Na + Ca 2+ Na + Net negative charge of the layers Ca 2+ Na + Ca 2+ Interlayer cations UMR CNRS 7016 LMPC

6 Amadeite, Amadeite, the origins to his its industrial applications Why using biopolymers extrated from seaweeds? Seaweeds UMR CNRS 7016 LMPC

7 Amadeite, Amadeite, the origins to his its industrial applications Why using clays and biopolymers extrated from seaweeds? Na + Ca 2+ Ca 2+ Na + Na + Ca 2+ + biopolymers montmorillonite Specific biding of big size mycotoxins such as Fumonisines, Trichothecenes and zearalenone UMR CNRS 7016 LMPC

8 CLAY ACTIVATION: THE P.I.L.C REVOLUTION Removal of the limiting factor Opening of the interlayer spaces allowing the entrance of the big size mycotoxins in the structure and their binding by adsorption on the internal surfaces. A wide spectrum of adsorption NO YES

9 Médical attentions préventives mesures CONTROL CONTROL management CONTROL housing CONTROL CONTROL Breeder feeding

10 MYCOTOXINS AND IMMUNITY constat Worldwide view : every country is concerned

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12 Contamination level worldwide

13 Occurrence of mycotoxins, some data: East Europe, Hungary Mycotoxin contamination in grain samples. Rafait et al, Feed additives and Contaminants

14 Occurrence of mycotoxins, some data: East Europe, Hungary Mycotoxin contamination in grain samples. Rafait et al, Feed additives and Contaminants

15 Mycotoxins doesn t end in the feed! Transferred into eggs, milk Decrease quality of food ( changes in lipid profiles, antioxidant concentrations ) Affect bloodcomposition JE Dvorska Veterinary Medicine department University Ukraine-WPC July 2008 Australia

16 Sources of mycotoxins Feed : plant + storage + in shed. ( 80% ) (Feed International 2008 Ref Guide) Airbone contamination ( 20%) (University Beijing International Animal Hygiene Conference Estonia) Bedding Litter ( Feed + airbone ) ( USDA Research Center Texas USA WPC 2008 )

17 Multiple toxiceffectsof mycotoxins Symptoms AFB1 DON T2 tox. FB1 OTA ZEA Anorexia Growth Liver damage Kidney damage Abortion Infertility Vulvovaginitis Pulmonary oedema Immunomodulation

18 Two ways of contamination AIRBORNE INHALATION FEED CONSUMPTION

19 Dynamism of the aerosol system MI CROORGANI SMS Feed partic les/mucus droplets form aerosols Endotoxins Mycotoxins Source: Stable gas and malodorous g as molecules bedding materiel, feces, condensation in the shed, feed. 19

20 Airborne contamination in dust Dominant microorganisms involved bacteria, fungi More damage with solid ammonia on a vector as dust than ammonia gaz Organic dust (feces, food, ) present higher risk of contamination than mineral dust (Guillam et al., 2007)

21 Airborne contamination by mycotoxin Mycotoxins can enter the birds 2 times, 3x, 4x, Ventilation Feed Faeces 2 3 Litter Condensa 1, 2, 3, 4 x

22 Airborne contamination by mycotoxin Example of contamination: «Mycotoxins in the ventilation systems 4 school in Finland» Mycotoxins Dust sample (ng/g = ppb) Aflatoxine B Deoxynivalenol (DON) Ochratoxin A T-2 toxin Zearalenone (ZEA) Imagine in poultry farming (Hintika et al., 2009)

23 Airborne: modulation of immune responsiveness of poultry ( WPC2008 Australia ) Important consequences of airborne on : Vaccine mangement Disease resistances Performances of industrial kept chickens HK Parmentier,G.De Vries, A Lammers, B Kemp Wageningen University Holland

24 The antigen-specific immune response FB1 (Taranu et al., 2005) Antigen DON (Pinton et al., 2008) Antigen Presenting Cells -- B lymphocytes T lymphocytes Antibody production +/ - Cytokines production +/ - Clonal expansion Cytotoxicity Humoral response Cellular response

25 DON and phagocytics cells DON 3500 control a LPS (10 µg/ml) IL -8 MIGRATION? CELLULAR COOPERATION? IL 8 concentration (pg/ml) d b c a,b,c,d (P <0.05) d d d d 0 DON (µm) 0 0 0, DON inhibits strongly the neutrophile IL 8 release (INRA/AFSSA, Waché et al., 2009)

26 Mechanism of action of DON DON inhibits protein synthesis For example, in hepatocyte (Doell et al., 2009): secreted protein (31 42 %) cellular protein (39-47 %)

27 TO DAY WE KNOW ONLY THE TOP OF THE ICEBERG ( nano )

28 In vitro tests Binding capacity of MTox Plus on Aflatoxins Contamination level 1 µg / mg of MTOX 2,5 µg / mg of MTOX 5 µg / mg of MTOX AF B1 AF B2 AF G1 AF G2 99, , ,5 99, Ref : LDA 22 France 2002 Trial protocol: Mise en solution dans un mélange KCl: 0,05 M HCl 0,013 M ( V/V ) àph 2,35 des différentes mycotoxines afin de réaliser une gamme de concentration à3 niveaux de l ordre de 10 à50 µg/ml par molécule. 110 mg de chaque support est mis en contact avec 1 ml ces différents niveaux de concentrations dans un tube conique. Après agitation au vortex, ces tubes sont placés àl étuve à38 +/ -1 C pendant 1 heure sous agitation par retournement Après remise à température ambiante, centrifugation à 1500 Tr/ mn pendant 5 mn Injection de 10 µl du surnageant dans un système de Chromatographie Liquide Haute Performance ( HPLC ) avec détection Fluorimétrique ( selon normes AFNOR ). Identification et Quantification des molécules retrouvées dans le surnageant

29 In vitro tests Binding capacity of MTox+ on DON and Zearalenone DON Intestinal adsorption % MTox + batch 1 59 MTox + batch 2 61 MTox + batch 3 65 Zearalenone Intestinal adsorption % MTox + batch 1 62 MTox + batch 2 72 MTox + batch 3 69 Ref : In vitro test, Utrecht University, NL, 2002

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32 Results presented at World Myctoxins Forum USA Fumonisin The bioaccessibility of fumonisin from contaminated pig feed was strongly inhibited by the addition of Amadéiteat the levels of 0.01% and 0.1%. A reduction of 50% to 60%of the bioaccessibility of fumonisin. Absorbed amount of fumonisin (ug) %-jej 0.01%-jej 0.10%-jej 0.00%-il 0.01%-il 0.10%-il 0.00%-total 0.01%-total 0.10%-total

33 Results presented at World Myctoxins Forum USA Absorbed amount of DON (ug) DON A strong inhibition of absorption of DON was found by the addition of Amadéiteat the level of 0.1%. The reduction 40% in comparison to the control %-jej 0.01%-jej 0.10%-jej 0.00%-il 0.01%-il 0.10%-il 0.00%-total 0.01%-total 0.10%-total

34 Montmorillonite Yeast cell walls Diatomaceous earth + + =

35 Spectrum of Activity AFLATOXINS OCHRATOXINS CLAYS ZON DON T-2 FUMONISIN YEAST CELL WALL

36 Aluminosilicates NOT ALL THE CLAYS ARE ALIKE

37 CLAY ACTIVATION: THE P.I.L.C REVOLUTION Removal of the limiting factor Opening of the interlayer spaces allowing the entrance of the big size mycotoxins in the structure and their binding by adsorption on the internal surfaces. A wide spectrum of adsorption NO YES MT.X+

38 Mycotoxins Take home messages

39 SIGNIFICANCE TO CHICKEN HEALTH OF THE IMMUNE IMPACT OF MYCOTOXINS Susceptibility to infectious disease Reactivation of chronic infection Vaccination efficacy Therapeutic efficacy

40 Mycotoxins synergy enemies N 1 High prevalence in feed Strong synergy with aflatoxine Strong effect on vaccine responses Immunity ppb Mycotoxins

41 The immune system of your animal your best partner keep it in a good shape silent but keep it

42 TRIAL RESULTS Asia Effects of MT.X+ inclusion in the diet on health and performance of broilers in Vietnam

43 TRIAL RESULTS Asia Effects of MT.X+ inclusion in the diet on health and performance of broilers in Vietnam

44 M TX+ AND LAYING HENS Malaysia Singapore months trial layers 2 sheds involved 1kg MT.X+ / ton ( = 40gr MT.X+/hen / year )

45 TRIAL RESULTS Asia Effects of MT.X+ on the performance of laying hens in Malaysia

46 M TX+ AND LAYING HENS Malaysia Singapore 2007 Cracked eggs reduced by 30% ( 1.1-0,75) Dirty eggs by 15% ( )

47 Why Zearalonone is phyto-oestrogenic

48 MTX+ in Hatchery Vaishnavi Breeding Farms Ltd, Dr.Ghuge S.B Director Warwati, Ambajogai, India.

49 MTX+ in Hatchery Reference with MT.x+ used in our farm. It s very please to say you that MT.x+ done very good job for saving our economical losses in breeders. We have got nice results :

50 MTX+ in Hatchery We observed that wrinkle eggs and shell-lesseggs come down Our hatchery eggs percentage also improved by 2.3%.

51 MTX+ in Hatchery In hatchability percentage : we improved our hatchability up to 2% We save cull percentage by 1.5% where as standard 2.0%. Cumulative hatchery performance improved by 3.5%.

52 M TX+ : Toxin binder New Generation Nanotechnology

53 AWARD - Malaysia 2007

54 THE IN & OUT ECO CONCEPT ABF 1,6 liter of water / bird must evaporate from the litter MAIN NH3 AND WATER EXCHANGE FOR A 1.9 KG BIRD Respiratory water g Evaporated Water from litter g NH3 23 g N 2 N 2 O Ingested water g Feed g 118 g N retained 60 g Water retained 1400g New litter Ex : straw 250 g/bird * = 38 g water + 1,5 g N Mistral Diaporama poultry 1005 * 20 birds / m2 de 1,9 kg [ C= 1,9 water / feed = 1,8 Faeces droppings 57 g N 1850 g water Chickens = 40 tons of droppings ~ 30T water + 36tons of water through respiration Litter losses Manure Per broiler g 275 g water 33 g N

55 Coccidiosis is increasingly being acknowledged as a major cause of diarrhoea in chickens. Oocysts in faeces spread the infection.

56 LITTER management & COCCIDIOSIS CONTROL

57 EasyApplication : feeders& drinkers

58 M FEED NANOTECHNOLGY FOR VECTORINGOF METALLIC IONS Mecanism of action : 3 ways

59 Property of M Feed Cu 2+ Cu 2+ Cu 2+ Structural copperat the end of the layer will act on the enzyme co-factor 59

60 M Feed 1 - DECREASES & CONTROLS VIABLE COUNTS OF INTESTINAL CLOSTRIDIUM, E COLI

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62 M Feed 2 - BOOSTS ENZYME ACTIVITY of gastrointestinal secretions

63 Enzymatic catalysis : Mecanism of action General information M FEED Cu 2+ Molecule for hydrolysis (A-B) co-factor co-factor addition (Cu 2+ ) Step 3 A-B co-factor Cu 2+ 63

64 Enzymatic catalysis : Mecanism of action enzymatic hydrolysis Final product after enzymatic hydrolysis (A and B) water Step 4 Step 5 A B Cu 2+ co-factor Cu 2+ co-factor co-factor regeneration 64

65 M FEED Results A -AMYLASE in small intestine B -2 times more amylase ( 18 versus 9 Somoguy units)

66 M FEED Results A -PROTEASE in small intestine B 25% increase with Cu MMT versus control or CuSO4

67 M FEED Results A -LIPASE in small intestine B 60% increase with Cu MMT versus control or CuSO4

68 M Feed 3 - IMPROVES INTESTINAL MUCOSAL MORPHOLOGY

69 A BLEND OF NATURAL FEED ADDITIVES (BNFAM FEED ) AS PERFORMANCE ENHANCER IN BROILERS Catherine G. Mangabat MS Animal Science University of the Philippines Los Baños A responsible and ethical feed industry for a sustainability in animal productions October 30, 2009 Hotel Windsor Suites, Bangkok Thailand

70 Effect of M FEED on villi height Villus height control Cu SO4 M FEED Duodenum 695b 702b 786a Jejinum 810b 821b 939a Ilium 585b 579b 681a

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72 IN CONCLUSION Mycotoxins under control No safety measure Economic losses

73 Climate changes = favor fungi Mycotoxins in feed : today the core problem, a situation we have to deal Not a disease with daily No choice, Need to adapt

74 Climate changes : need to adapt

75 Thanks for your attention