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August 4th, 2012
Daniel Petri 1 and Daniel Karunakaran 2
1 Animal & Environmental Applications, Dupont Nutrition & Health,
W227N752 Westmound Drive, Waukesha, WI 53186. 2 Danisco Animal Nutrition, DuPont Industrial Biosciences,
2008 S 8th St, St. Louis, MO 63104
Bacterial Relationships in Enteric Disease
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2 8/4/2012
~1e10 cfu/g
~1e5 cfu/g
~1e6 –
1e10 cfu/g ~1e11 cfu/g
Adapted from
Siragusa et al. 2010
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Mucosa differences along the GIT
3 Atmua et al. 2001
Page 4
4
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
d14 no DFM d14 DFM d28 no DFM d28 DFM d42 no DFM d42 DFM
Effects of DFM SupplementationMajor genera (> 0.1%)
p = 0.037 p = 0.425 p = 0.645
Succession of gut microbiota in healthy broiler chickens
Neumann et al., 2011
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Rough overview of major GIT microbes in the intestine
5
Bacterial species Beneficial Harmful Growth Rate Degrades Produces Sporeformer
LAB VFA (eg group D Streptococcus)
some (eg group A Streptococcus)
intermediate CHO lactate no
Enterococcus VFA, immuno-
modulation (E. facium)
eg E. cecorum (antibiotic resistence)
intermediate CHO lactate no
Lactobacillus Adherent, Immu-no modulation,
E. coli antagonist - intermediate CHO lactate no
Bifidobacterium Adherent in SI, VFA - slow lactate butyrate no
Propionibacterium VFA - slow CHO propionate no
E. coli if non-virulent (E. coli Nissle)
APEC (adherent, toxins)
fast CHO acetate, LPS no
Clostridium
C. cluster IV - soluble fiber butyrate yes
C. cluster XIVa - insoluble
fiber butyrate yes
- C. cluster I
(C. perfringens) fast
mucus, proteins
entero- toxins, H2S
yes
Bacteroides, Prevotella
Neutral slow fiber acetate, LPS no
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A balanced immune system is necessary for healthy birds
Inflammation is needed to fight
pathogens
Excessive inflammation decreases
performance
Direct-fed microbials (DFM) can be
used to influence the immune
system
Nature Reviews Immunology, 2009
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8/4/2012 7
Asymptomatic GD Symptomatic .
Hibberd et al., 2008
GD in Broiler Chickens
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Ileum mucosa microbiota of 11 live capture birds
Principle Component Analysis
8/4/2012 8
Asymptomatic birds
GD affected birds
Early et al., 2006
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GD in Turkeys - Relative abundance of major MspI TRFs
8/4/2012 9
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Composition of clostridial spores recovered from GD endemic and non-endemic turkey grow-out litter
9%0%
51%5%
6%
8%
2%
2%
0% 3%
0%
1%
3%9%
Clostridial Spores in Grow-Finish Litter
C. tertium C. septicum C. perfringens
C. beijerinckii C. butyricum C. paraputrificum
C. cadaveris C. novyi C. sporogenes
C. cochlearium C. tetani C. sordellii
C. difficile Others
21%
10%
2%6%
22%1%
24%
6%
6%
Clostridial Spores in Brood Litter
C. tertium C. perfringens C. beijerinckii
C. butyricum C. paraputrificum C. cochlearium
C. difficile C. innocuum Others
49%
5%
16%
2%
12%
4%
12%
1%
Enriched Spores in Brood Litter
C. tertium C. butyricum C. paraputrificum
C. novyi C. sporogenes C. cochlearium
C. bifermentans others
4%
3% 1%
4%
8%
8%
4%
23%
43%
3%
Enriched Spores in Grow-Finish Litter
C. tertium C. perfringens C. beijerinckii
C. butyricum C. paraputrificum C. cadaveris
C. novyi C. sporogenes C. cochlearium
C. sordellii
15%
40%
4%
14%
15%
1%1%
1%
4%4%
Clostridial Spores in Grow-Finish Litter from Non-Endemic GD farms
C. tertium C. perfringens C. beijerinckii
C. butyricum C. paraputrificum C. cadaveris
C. cochlearium C. tetani C. difficile
Others
7%
1%
57%5%2%
3%
4%
3%
1% 4%
1%1% 12%
Clostridial Spores in Grow-Finish Litter from Endemic GD farms
C. tertium C. septicum C. perfringens
C. beijerinckii C. butyricum C. paraputrificum
C. cadaveris C. novyi C. sporogenes
C. cochlearium C. sordellii C. difficile
Others
7%
1%
57%5%2%
3%
4%
3% 1%
4%0%
0% 1%
1%
0%12%
Chart Title
C. tertium C. septicum C. perfringens C. beijerinckii C. butyricum C. paraputrificum
C. cadaveris C. novyi C. sporogenes C. cochlearium C. tetani C. bifermentans
C. sordellii C. difficile C. innocuum Others
Brood Grow-out
GD Nonendemic GD Endemic
Enriched Brood Enriched Grow-out
Clostridial Spores Cultured from Litter and Chopped Meat
Enrichments
9%0%
51%5%
6%
8%
2%
2%
0% 3%
0%
1%
3%9%
Clostridial Spores in Grow-Finish Litter
C. tertium C. septicum C. perfringens
C. beijerinckii C. butyricum C. paraputrificum
C. cadaveris C. novyi C. sporogenes
C. cochlearium C. tetani C. sordellii
C. difficile Others
21%
10%
2%6%
22%1%
24%
6%
6%
Clostridial Spores in Brood Litter
C. tertium C. perfringens C. beijerinckii
C. butyricum C. paraputrificum C. cochlearium
C. difficile C. innocuum Others
49%
5%
16%
2%
12%
4%
12%
1%
Enriched Spores in Brood Litter
C. tertium C. butyricum C. paraputrificum
C. novyi C. sporogenes C. cochlearium
C. bifermentans others
4%
3% 1%
4%
8%
8%
4%
23%
43%
3%
Enriched Spores in Grow-Finish Litter
C. tertium C. perfringens C. beijerinckii
C. butyricum C. paraputrificum C. cadaveris
C. novyi C. sporogenes C. cochlearium
C. sordellii
15%
40%
4%
14%
15%
1%1%
1%
4%4%
Clostridial Spores in Grow-Finish Litter from Non-Endemic GD farms
C. tertium C. perfringens C. beijerinckii
C. butyricum C. paraputrificum C. cadaveris
C. cochlearium C. tetani C. difficile
Others
7%
1%
57%5%2%
3%
4%
3%
1% 4%
1%1% 12%
Clostridial Spores in Grow-Finish Litter from Endemic GD farms
C. tertium C. septicum C. perfringens
C. beijerinckii C. butyricum C. paraputrificum
C. cadaveris C. novyi C. sporogenes
C. cochlearium C. sordellii C. difficile
Others
7%
1%
57%5%2%
3%
4%
3% 1%
4%0%
0% 1%
1%
0%12%
Chart Title
C. tertium C. septicum C. perfringens C. beijerinckii C. butyricum C. paraputrificum
C. cadaveris C. novyi C. sporogenes C. cochlearium C. tetani C. bifermentans
C. sordellii C. difficile C. innocuum Others
Brood Grow-out
GD Nonendemic GD Endemic
Enriched Brood Enriched Grow-out
Clostridial Spores Cultured from Litter and Chopped Meat
Enrichments
7%
1%
57%5%2%
3%
4%
3% 1%
4%0%
0% 1%
1%
0%12%
Chart Title
C. tertium C. septicum C. perfringens C. beijerinckii C. butyricum C. paraputrificum
C. cadaveris C. novyi C. sporogenes C. cochlearium C. tetani C. bifermentans
C. sordellii C. difficile C. innocuum Others
Non-endemic (n= 73) Endemic (n= 136)
Early et al., 2006
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C. perfringens and C. septicum
in Turkey Cellulites
11
Neuman et al., 2007a,b
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Diversity of Avian Pathogenic E. coli
Toxin Genes in Pullets vs. Laying Hens
12
8
2 2
0 0.40 0.3
8
11
5
3
12
3
36
4
2
9
2211
2
9
43 3
20
13
17
10
15
0
4
8
12
16
20
24
28
32
36 Pullets Layers
Iss/i
ucC
/tsh
/cvaC
iss/cvaC
cvaC
Iss
/cv
aC
cvaC
no
ge
ne
s
no
ge
ne
s
Iss
/ts
h/c
va
C
Iss
/ts
h/c
va
C
Iss
/iu
cC
/tsh
/cv
aC
iucC
/tsh
iuc
C/t
sh
tsh
/cv
aC
tsh
/cv
aC
Batzley & Rehberger, 2006 42% Non-APEC 27%
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Characterization of the duodenal microbiota of commercial
layer hens affected by Focal Duodenal Necrosis
8/4/2012 13
Benson et al., 2009
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Characterization of the duodenal microbiota of commercial
layer hens affected by Focal Duodenal Necrosis
8/4/2012 14
Benson et al., 2009
Page 15
Salmonella infection
15
Atm
ua
et
al. 2
00
1
Page 17
Summary
In enteric disease, healthy gut microbiota balance is disrupted
Increased abundance of proteolytic and/or toxin producing Clostridia,
specifically C. perfringens, is directly or indirectly associated with most enteric
diseases
C. perfringens shows high variability on the species level leading to various
diseases (NE, GD)
Enteric disease via Avian Pathogenic E. coli is dependent on number of toxin
genes present
Enteric Salmonella infection is dependent on previous intestinal disease
Acute enteric disease is usually accompanied by a decrease in beneficial
bacterial populations, for example Lactobacillus and group D Streptococcus
8/4/2012 17
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