prebiotics and probiotics in animal production: present status and future perspectives ·...

Prebiotics and probiotics in animal production: present status and future

perspectives

R. Ducatelle, F. Haesebrouck and F. Van Immerseel

Department of Pathology, Bacteriology and Avian Medicine

Faculty of Veterinary Medicine

We are born 100% human

We are born 100% human but we die 100% bacterial

We are born 100% human but we die 100% bacterial in between we are 90% bacterial

105/g

101/g

103/g

108/g

1011/g

1011/g

1. It is forbidden by law in the EU to use antibiotics for treatment of Salmonella in poultry

2. It is forbidden by law in the EU to use antibiotics as growth promoters in farm animals

Probiotics:

Single or mixed cultures of living microorganisms which beneficially Affect the host by improving the properties of the indigenous microbiota. (Fuller, 1992)

Prebiotics:

Non-digestible feed ingredients that selectively favor the multiplication or metabolic activity of a specific fraction of the intestinal microbiota. (Gibson & Roberfroid, 1995)

>100 microorganisms >30 gut flora stabilizers No separate category for prebiotics

Probiotics:

Single or mixed cultures of living microorganisms which beneficially affect the host by improving the properties of the indigenous microbiota. (Fuller, 1992)

Enterococcus, Lactobacillus, Saccharomyces, Bacillus, Pediococcus

Prebiotics:

Non-digestible feed ingredients that selectively favor the multiplication or metabolic activity of a specific fraction of the intestinal microbiota. (Gibson & Roberfroid, 1995)

Probiotics improve performance under challenge conditions: Coccidiosis Clostridium perfringens Salmonella LPS (acute phase response)

Probiotics, their health benefits and applications for developing healthier foods: a review

FEMS Microbiology Letters Volume 334, Issue 1, pages 1-15, 28 MAY 2012 DOI: 10.1111/j.1574-6968.2012.02593.x http://onlinelibrary.wiley.com/doi/10.1111/j.1574-6968.2012.02593.x/full#fml2593-fig-0001

(Nagpal et al., 2012)

Probiotics improve performance under challenge conditions: Coccidiosis Clostridium perfringens Salmonella LPS (acute phase response)

Probiotics inhibit bacterial translocation: Wire-floor model (Wideman et al., Poult. Sci., 2012)

Probiotics:

Single or mixed cultures of living microorganisms which beneficially affect the host by improving the properties of the indigenous microbiota. (Fuller, 1992)

Prebiotics:

Non-digestible feed ingredients that selectively favor the multiplication or metabolic activity of a specific fraction of the intestinal microbiota. (Gibson & Roberfroid, 1995)

FOS, XOS, GOS, IMO, RFO

Prebiotics : Increase bioavailability of minerals Stimulate the immune system

Probiotics:

Single or mixed cultures of living microorganisms which beneficially affect the host by improving the properties of the indigenous microbiota. (Fuller, 1992)

Prebiotics:

Non-digestible feed ingredients that selectively favor the multiplication or metabolic activity of a specific fraction of the intestinal microbiota. (Gibson & Roberfroid, 1995)

FOS, XOS, GOS, IMO, RFO

MOS:

Experimental model 1

Experimental set up

- Antimicrobial growth promoter

(Zn-bacitracin 100 mg/kg)

- 2 ≠ carbohydrate sources

↔

Corn Wheat/rye

(Teirlynck et al., Brit. J. Nutr. 2009)

Feed conversion

Feed conversion d1-15

1,24

1,26

1,28

1,3

1,32

1,34

1,36

1,38

1,4

1,42

Diet

Feed

co

nvers

ion

W/R

W/R+B

M

M+B

Villus length

Villi Duodenum M+B diet (dag 15) Villi Duodenum W/R diet (dag 15)

Villus length

Villus length in duodenum

1500

1600

1700

1800

1900

2000

2100

2200

2300

2400

2500

1 2 3

Age (days)

Villu

s len

gth

(u

m)

W/ R

W/ R + B

M

M + B

d15 d29 d42

Villus length in duodenum

1500

1600

1700

1800

1900

2000

2100

2200

2300

2400

2500

day 15 day 29 day 42

Age(days)

Villu

s len

gth

(u

m)

W/ R

W/ R + B

M

M + B

d15 d29 d42

Villus fusion

Villi Jejunum M+B dieet (dag 15)

Villi Jejunum W/R dieet (dag 15) Villi Jejunum W/R dieet (dag 15)

Villus fusion

Villusfusion jejunum d29

0

1

2

3

4

5

6

Dieet

Sco

re

W/R W/R+B M M+B

Muscularis thickness

M+B: Dikte tunica muscularis duodenum (dag 15) (20x)

W/R: Dikte tunica muscularis duodenum (dag 15) (20x)

Muscularis thickness

Thickness tunica muscularis duodenum

140

190

240

290

340

390

day 15 day 29 day 42

age (days)

Dik

te (

um

) W/R

W/R + B

M

M + B

M+B: T-cel infiltration in duodenum

W/R: T-cel infiltration in duodenum

M+B: T-cel infiltration in caecum W/R: T-cel infiltration in caecum

Immune cell infiltration

T-lympocyte infiltration d15

4

6

8

10

12

14

16

18

20

22

place

T-l

ym

ph

ocyte

are

a %

W/ R

W/ R + B

M

M + B

duo jej ile cae

T-lymphocyte infiltration d15

4

9

14

19

24

T-ly

mph

ocyt

e ar

ea %

W/R

W/R + B

M

M + B

duo jej ile cae

Goblet cells

W/R: ileum (100 x) M: ileum (100x)

Results

Protective mucin layer

Results t-RFLP

Effect of diet

Green: wheat/rye

Blue: wheat/rye + Zn- Bacitracine

Yellow: maize

Red: Maize + Zn-Bacitracine

2 weeks old

Statement 1: Growth promotors cause a shift in the microbiota of caeca and colon

Statement 1: Growth promoters cause a shift in the microbiota of caeca and colon Statement 2: Growth promoters promote shift from inflammation to oral tolerance

Experimental model 2

High molecular weight pectins

Langhout et al., Poultry Sci. 2000

Starter Grower Finisher

Cellulose d0-d13

%

Pectine d0-d13

%

Cellulose d13-d26

%

Pectine d13-d26

%

Cellulose d26-d39

%

Pectine d26-d39

%

Corn 47,35 47,35 48,64 48,64 50,90 50,90

Soybean meal 48 (46) 31,95 31,95 29,60 29,60 27,44 27,44

Soybeans 4,68 4,68 4,68 4,68 4,68 4,68

Animal fat 3,60 3,60 5,45 5,45 5,00 5,00

Soybean oil 1,85 1,85 1,45 1,45 2,04 2,04

CaCO3 0,33 0,33 0,27 0,27 0,27 0,27

D-Ca-phosphate 1,81 1,81 1,60 1,60 1,30 1,30

NaCl 0,22 0,22 0,24 0,24 0,26 0,26

NaHCO3 0,14 0,14 0,11 0,11 0,10 0,10

L-Lys HCl 0,205 0,205 0,134 0,134 0,174 0,174

DL-methionine 0,295 0,295 0,236 0,236 0,237 0,237

L-threonine 0,050 0,050 0,025 0,025 0,025 0,025

Vit. & Tr. el. Premix 1,00 1,00 1,05 1,05 1,05 1,05

Phytase 0,02 0,02 0,02 0,02 0,02 0,02

Cellulose 6,50 3,50 6,50 3,50 6,50 3,50

Pectines 0,00 3,00 0,00 3,00 0,00 3,00

NSP (Bio FW) no no no no no no

Sum 100,00 100,00 100,01 100,01 100,00 100,00

Feed composition

Performance parameters

0

200

400

600

800

1000

1200

ggd13 ggd26

0

10

20

30

40

50

60

70

Vd0-13 Vd0-26

0,0

0,5

1,0

1,5

2,0

2,5

VOd0-13 VOd0-26

Weight

Feed intake

Feed conversion

gew

ich

t (g

)

voe

r/d

ier/

dag

(g)

VC

R

338a

251b

954a

733b

30,8a

27b

62,5a 59,4a

1,34a

1,66b 1,78a

2,23b

cellu

lose

cellu

lose

cellu

lose

pec

tin

e

pec

tin

e

pe

ctin

e

pec

tin

e

pec

tin

e

pec

tin

e

cellu

lose

cellu

lose

cellu

lose

Gemiddeldes met verschillend superscript zijn significant verschillend (P = 0,001)

day 22

Dysbacteriosis scoring

Cranial

Caudal

Cage

Treatment

Chicken

Gut Ballooning

Inflamma-

tion Vessels

Flaccid

Content

Thickness Translucen

cy Fragility

Inflamma-

tion Vessels

Flaccid

Content

Thickness Translucen

cy Fragility

Un- digested

feed

33 cellulose 1 1 0 0 1 0 0 0 0 0 0

35 pectine 1 0 0 0 1 0 0 0 1 1 0

Group Score Stdv

Cellulose 2,12 0,946

Pectine 3,54 0,997

0

2

4

6

8

10

12

14

Aan

tal d

iere

n

Cellulose Pectine 0 1

1

2

2

3

3

4

4

5

3 animals/pen 24animals/group

(according to Teilynck et al., Avian Pathol. 2011)

day 22

Ileum (relative percentage)

Caecum (relative percentage)

0%

2%

4%

6%

8%

10%

12%

14%

16%

18%

20%

cellulose pectine

Enterobacteriaceae

Cluster I

cluster IV

8,0 %

3,32 %

0,48 %

14,12 %

1,03 % 0,17 %

0%

2%

4%

6%

8%

10%

12%

14%

16%

18%

cellulose pectine

B. pullicaecorum

Enterobacteriaceae

Lactobacilli

Cluster I

CoA-transferase

11,31 %

2,3 %

3,1 %

1,3 % 0,9 %

14,1 %

13,3 % 12,6 %

0,9 % 0,5 %

Butyrate production

butyryl-CoA:acetate CoA-transferase

Butyryl CoA

Butyrate

Acetate

Acetyl-CoA

day 22

ButCoA-AcCoA-transferase

Log

cfu

/g f

aece

s

6

6,5

7

7,5

8

8,5

9

9,5

10

10,5

0 1 2 3 4 5 6

cellulose

pectine

Dysbacteriosis score

day 22

CoA-transferase

Log

cfu

/g f

aece

s

6

6,5

7

7,5

8

8,5

9

9,5

10

10,5

0 1 2 3 4 5 6

cellulose

pectine

Dysbacteriosis score

Butyric acid

Energy source for colonocytes

Anti-inflammatory properties Anticarcinogenic potential

What is the effect of SCFA on Salmonella?

Five groups of 20 chickens :

CTRL FORMIC ACETIC PROPIONIC BUTYRIC

Infection at day 5 with 5.10³ cfu S. Enteritidis

Euthanasia at day 8

Bacteriological analysis of caeca, liver and spleen : titration on BGA

ALL ACIDS IN COATED FORM

(Van Immerseel et al., Poultry Sci. 2004)

Caecal colonization of Salmonella

2

3

4

5

6

7

8

CTRL FOR ACE BUT PROP

treatment

mean log c

fu/g

a

a

b

c

ac

Thank you for your attention

Faculty of Veterinary Medicine Ghent University