bacterial relationships in enteric...

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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

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. butyricum C. paraputrificum C. cadaveris


C. sordellii

15%

40%

4%

14%

15%

1%1%

1%

4%4%

Clostridial Spores in Grow-Finish Litter from Non-Endemic GD farms



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. 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. cochlearium C. tetani C. sordellii

C. difficile Others

21%

10%

2%6%

22%1%

24%

6%

6%

Clostridial Spores in Brood Litter


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. bifermentans others

4%

3% 1%

4%

8%

8%

4%

23%

43%

3%

Enriched Spores in Grow-Finish Litter




C. sordellii

15%

40%

4%

14%

15%

1%1%

1%

4%4%

Clostridial Spores in Grow-Finish Litter from Non-Endemic GD farms



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. cochlearium C. sordellii C. difficile

Others

7%

1%

57%5%2%

3%

4%

3% 1%

4%0%

0% 1%

1%

0%12%

Chart Title




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




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

Salmonella infection

15

Atm

ua

et

al. 2

00

1

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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