Sécurité sanitaire en élevage

avicole en climat chaud :

quelles stratégies de

prévention ?

• Alain Réocreux September 2014

• Olmix France

Nanotechnology applications

in

Animal Feeds

Eco Concept

• M T.X+ Toxin Binder

• M Feed Natural Growth Promoter

LMPC

UMRCNRS 7016

ab

c

Amadeite®, the origins to his industrial applications

Discovered in 2005 by Olmix in France

LMPC

UMRCNRS 7016

Collaboration between Olmix, LPMC and CEVA

LMPC

UMRCNRS 7016

LMPC

UMRCNRS 7016

Why using Montmorillonite?

ab

c

Na+

Na+

Ca2+

Ca2+

Na+

Ca2+

oxygen

silicon

aluminium and

magnesium

hydroxyl or

oxygen

Net negative charge of the layers

Interlayer cations

Amadeite™, the origins to his industrial applicationsAmadeite® , the origins to its industrial applications

Why using biopolymers extrated from seaweeds?

LMPC

UMRCNRS 7016

Seaweeds

Amadeite™, the origins to his industrial applicationsAmadeite® , the origins to its industrial applications

Specific biding of big size mycotoxins such as Fumon isines, Trichothecenesand zearalenone

Amadeite™, the origins to his industrial applications

LMPC

UMRCNRS 7016

LMPC

UMRCNRS 7016

montmorillonite

Na+

Na+

Ca2+

Ca2+

Na+

Ca2+

+

Why using clays and biopolymers extrated from seaweeds?

biopolymers

Amadeite® , the origins to its industrial applications

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.

Removal of the limiting factor

NO

YES

A wide spectrum of adsorption

CLAY ACTIVATION: THE P.I.L.C REVOLUTIONCLAY ACTIVATION: THE P.I.L.C REVOLUTION

housing feeding

Médical attentions préventives mesures

management

Breeder

CONTROL

CONTROL

CONTROL

CONTROL

CONTROL

feeding

MYCOTOXINS AND IMMUNITY

constat

Worldwide view : every country is concerned

Contamination level worldwide

Occurrence of mycotoxins, some data:

East Europe, Hungary

Mycotoxin contamination in grain samples. Rafait et a l, 2000. Feed additives and Contaminants

Occurrence of mycotoxins, some data:

East Europe, Hungary

Mycotoxin contamination in grain samples. Rafait et a l, 2000. Feed additives and Contaminants

Mycotoxins doesn’t end in the feed!

• Transferred into eggs, milk …

• Decrease quality of food ( changes in lipid profiles, antioxidant

concentrations…)

• Affect blood composition

JE Dvorska Veterinary Medicine department University Ukraine-WPC July 2008 Australia

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 )

Multiple toxic effects of mycotoxins

Symptoms AFB1 DON T2 tox. FB1 OTA ZEA

Anorexia + +++ +++ + + 9

Growth +++ +++ ++ + + 10

Liver damage +++ ++ + 6

Kidney damage + +++ 4

Abortion + ++ 3

Infertility +++ 3

Vulvovaginitis +++ 3

Pulmonary oedema +++ 3

Immunomodulation +++ ++ ++ +++ + + 12

10 8 7 12 7 9

Two ways of contamination Two ways of contamination

AIRBORNE INHALATION

FEED CONSUMPTION

19

Dynamism of the aerosol systemDynamism of the aerosol system

MICROORGANI SMS

E ndotoxins

MycotoxinsStable gas and malodorous

g as molecules

F eed partic les /mucus

droplets form aerosols

Source:

bedding materiel, feces, condensation in the shed, feed.

• 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)

Airborne contamination in dustAirborne contamination in dust

Airborne contamination by mycotoxinAirborne contamination by mycotoxin

Mycotoxins can enter the birds 2 times, 3x, 4x,…

1, 2, 3, 4 x…

1

Feed

Ventilation

4

Faeces2

3

Litter

Condensation

5

Mycotoxins Dust sample (ng/g = ppb)

Aflatoxine B1 18 - 60

Deoxynivalenol (DON) 120 – 390

Ochratoxin A 24 - 78

T-2 toxin 18 - 60

Zearalenone (ZEA) 42 - 140

Airborne contamination by mycotoxinAirborne contamination by mycotoxin

Example of contamination:

« Mycotoxins in the ventilation systems 4 school in Finland »

(Hintika et al., 2009)

…Imagine in poultry farming…

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

Cytokines

production

Cytotoxicity

Cellular response

Clonal

expansion

The antigenThe antigen--specific immune responsespecific immune response

Antigen

Antibody

production

Humoral response

Antigen Presenting Cells

B lymphocytes T lymphocytes

+/

-

+/

-

FB1 (Taranu et al., 2005)

--

DON (Pinton et al., 2008)

DON

MIGRATION?

DON and phagocytics cellsDON and phagocytics cells

(INRA/AFSSA, Waché et al., 2009)

IL - 8

CELLULAR

COOPERATION?

DON inhibits strongly the neutrophile IL –8 release

a,b,c,d (P <0.05)

d

a

c

dd d d

b

IL –

8 co

ncen

trat

ion

(pg/

ml)

DON (µM) 0 0 0,5 1 2 5 10 50

0

500

1000

1500

2000

2500

3000

3500

control LPS (10 µg/ml)

DON inhibits protein synthesis

Mechanism of action of DONMechanism of action of DON

For example, in hepatocyte (Doell et al., 2009):

• secreted protein (31 – 42 %)

• cellular protein (39 - 47 %)

TO DAY WE KNOW ONLY THE TOP OF THE

ICEBERG ( nano )

Contamination level

AF B1 AF B2 AF G1 AF G2

1 µg / mg of MTOX

99,5 97 97 96,5

2,5 µg / mg of MTOX

100 98,5 99,5 98

5 µg / mg of MTOX

99 95 98 94

In vitro tests

Trial protocol :Mise en solution dans un mélange KCl : 0,05 M – HCl 0,013 M ( V/V ) à pH 2,35 des différentes mycotoxines afin de réaliser une gamme de concentration à 3 niveaux de l’ordre de 10 à 50 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 à l’étuve à 38 + / - 1°C pendant 1 heure sous agitation par retournementAprès remise à température ambiante, centrifugation à 1500 Tr / mn pendant 5 mnInjection 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

Ref : LDA 22

France 2002

Binding capacity of MTox Plus on Aflatoxins

In vitro tests

Binding capacity of MTox+ on DON and Zearalenone

Ref : In vitro test, Utrecht University, NL, 2002

ZearalenoneIntestinal adsorption %

MTox + batch 1 62MTox + batch 2 72MTox + batch 3 69

DONIntestinal adsorption %

MTox + batch 1 59MTox + batch 2 61MTox + batch 3 65

Results presented at World Myctoxins

Forum USA

Fumonisin

The bioaccessibility of fumonisin

from contaminated pig feed was

strongly inhibited by the

addition of Amadéite at the

levels of 0.01% and 0.1%.

A reduction of 50% to 60% of

the bioaccessibility of

fumonisin. 0

10

20

30

40

50

60

70

0.00%

-jej

0.01

%-je

j0.

10%

-jej

0.00

%-il

0.01

%-il

0.10%

-il

0.00

%-to

tal

0.01

%-to

tal

0.10

%-to

tal

Abs

orbe

d am

ount

of f

umon

isin

(ug)

Results presented at World Myctoxins

Forum USA

0

5

10

15

20

25

0.00

%-je

j0.

01%-je

j0.

10%

-jej

0.00

%-il

0.01%

-il0.

10%

-il

0.00

%-to

tal

0.01

%-to

tal

0.10

%-to

tal

Abs

orbe

d am

ount

of D

ON

(ug)DON

A strong inhibition of absorption

of DON was found by the addition

of Amadéite at the level of 0.1%.

The reduction 40% in comparison

to the control.

Yeast cell walls Diatomaceous earthMontmorillonite

+ +

=

Spectrum of Activity

AFLATOXINS

OCHRATOXINS

ZON

DON

FUMONISIN

T-2

CLAYS

YEAST CELL WALL

Aluminosilicates

• NOT ALL THE CLAYS ARE ALIKE

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.

Removal of the limiting factor

NO

YES

MT.X+

A wide spectrum of adsorption

CLAY ACTIVATION: THE P.I.L.C REVOLUTIONCLAY ACTIVATION: THE P.I.L.C REVOLUTION

Mycotoxins

Take home messages

SIGNIFICANCE TO CHICKEN HEALTH

OF THE IMMUNE IMPACT OF MYCOTOXINS

Susceptibility to infectious disease

Reactivation of chronic infection

Vaccination efficacy

Therapeutic efficacy

Mycotoxins synergy

enemies N°1

ImmunityMycotoxins

ppb

• High prevalence in feed

• Strong synergy with aflatoxine

• Strong effect on vaccine responses

The immune system of your animal

your best partner

keep it in a good shape

but keep it silent

Asia

TRIAL RESULTS

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

Asia

TRIAL RESULTS

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

M TX+ AND LAYING HENSMalaysia Singapore 2007

• 6 months trial

• 100 000 layers

• 2 sheds involved

• 1kg MT.X+ / ton ( = 40gr MT.X+/hen / year )

Asia

TRIAL RESULTS

Effects of MT.X+ on the performance of laying hens in Malaysia - 2007

M TX+ AND LAYING HENSMalaysia Singapore 2007

• Cracked eggs reduced by 30% ( 1.1 - 0,75)

• Dirty eggs by 15% ( 10.87 9.22)

Why Zearalonone is phyto-oestrogenic

MTX+ in Hatchery

• Vaishnavi Breeding Farms Ltd, Dr.Ghuge S.B Director

• Warwati, Ambajogai,

• India.

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 :

MTX+ in Hatchery

•We observed that wrinkle eggs and

shell-less eggs come down

• Our hatchery eggs percentage also

improved by 2.3%.

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%.

M TX+ : Toxin binder

New Generation Nanotechnology

AWARD - Malaysia 2007

THE IN & OUT ECO CONCEPT ABF

Respiratory water 3 300 g

Ingested water 6 500 g

Evaporated Water from litter

1 600 g NH3 23 g

N2 N2O

Feed 3 600 g 118 g

N retained 60 g Water retained 1400g

Faeces droppings 57 g N

1850 g water Litter losses

Manure Per broiler

1 100 g 275 g water

33 g N New litter Ex : straw 250 g/bird * = 38 g water + 1,5 g N

1,6 liter of water / bird must evaporate from the litter

MAIN NH3 AND WATER EXCHANGE FOR A 1.9 KG BIRD

* 20 birds / m2 de 1,9 kg [ C= 1,9 water / feed = 1,8

20 000 Chickens = 40 tons of droppings ~ 30T water

+ 36tons of water through respirationMistral Diaporama poultry 1005

Oocysts in faeces spread the infection.

Coccidiosis is increasingly being acknowledged as a major cause of diarrhoea in chickens.

LITTER management & LITTER management & COCCIDIOSIS CONTROLCOCCIDIOSIS CONTROL

Easy Application : feeders& drinkers

M FEED NANOTECHNOLGY

FOR VECTORING OF METALLIC IONS

Mecanism of action : 3 ways

59

Cu 2+

Cu 2+

Property of M Feed

Structural copperat the end of the layer will act on the enzyme co-factor

Cu2+

M Feed

1 - DECREASES & CONTROLS VIABLE

COUNTS OF INTESTINAL

CLOSTRIDIUM , E COLI

M Feed

2 - BOOSTS ENZYME ACTIVITY of

gastrointestinal secretions

63

Enzymatic catalysis : Mecanism of action

Cu2+ co-factor

M FEED

co-factor

Step 3

Molecule for hydrolysis

A-B(A-B)

General information

co-factor addition (Cu 2+)

Cu2+

64

co-factor

Step 4

water

Step 5

co-factor

Final product after enzymatic hydrolysis

(A and B)

AB

co-factor regeneration

Cu2+Cu2+

enzymatic hydrolysis

Enzymatic catalysis : Mecanism of action

M FEED Results

• A - AMYLASE in small intestine

• B - 2 times more amylase ( 18 versus 9 Somoguy units)

M FEED Results

• A - PROTEASE in small intestine

• B 25% increase with Cu MMT versus control or CuSO4

M FEED Results

• A - LIPASE in small intestine

• B 60% increase with Cu MMT versus control or CuSO4

M Feed

3 - IMPROVES INTESTINAL

MUCOSAL MORPHOLOGY

A BLEND OF NATURAL FEED ADDITIVES (BNFAM FEED ) AS

PERFORMANCEENHANCER 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, 2009Hotel Windsor Suites, Bangkok Thailand

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

IN CONCLUSION

Mycotoxins under control

No safety measure

Economic losses

Climate changes = favor fungi

•Mycotoxins in feed : today the core problem

– Not a disease , a situation we have to deal with daily

No choice , Need to adapt

Climate changes : need to adapt

Thanks for your attention