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