Prestage Department of Poultry Science

Developing Leadership for the Global Marketplace

The Role of Intestinal Health in
Broiler Production
Edgar O. Oviedo-Rondón
DVM, MSc., PhD, Dipl. ACPV
Associate Professor / Extension Specialist
 Outline of this Presentation
● Importance of intestinal health
● Gut microbiome and its impacts
● Factors that disrupt gut ecosystem affecting health
● Current methods to study microbial ecology
● Alternatives to growth promoting antibiotics
Healthy Guts: Key for Poultry Production
 Factors Giving More Relevance to
Gut Health Lately
● Growth promotant antibiotic bans or avoidance
● Increased concern for food safety – microbiological
● Coccidiosis: increased resistance & lack new products
● Growing environmental and welfare concerns
● Feed ingredient costs & increased use of byproducts
● Higher variability on feed composition
● Need to improve digestibility and efficiency
Indirect

Indirectly
Tendencies in the
Poultry Industry
 Outline of this Presentation
● Importance of intestinal health
● Gut microbiome and its impacts
 Lactobacilos, Lactobacilos,
Streptococos & Streptococos &
Coliformes Coliformes

Bifidobactérias,
Bacteroides,
Streptococos &
Clostridium

Gut: Ecosystem with bird as host

Gut Ecosystem in Equilibrium = Healthy
 Effects of Microbial Populations in the Intestine
11 Edgar

Negative Positive
Oviedo

Bacteroides
Toxin production, potential
carcinogens = AGV, Amonia Bifidobacterium

Intestinal putrefaction = Eubacterium Inhibition of pathogenic

biogenic amines Metanogenic bacteria
Transformation de anaer. G(+) cocci Immune system
Bile salts stimulation
Sulfate reducers
8 Lactobacillus
Diarrhea E. coli Help in digestion and
Constipation
enterobacteria absorption of nutrients
Infections
and minerals
Systemic effects
Veillonella
Vitamin Synthesis
Clostridium
Staphylococcus
Salmonella Vibrionaceae
Ps. aeruginosa 2 (G.R Gibson, 1998)
Pathobionts Populations, log cfu/g fezes
 Microbial Populations in the Intestine
11 Edgar

Negative Positive
Oviedo

Bacteroides
Toxin production, potential
carcinogens = AGV, Amonia Bifidobacterium

Intestinal putrefaction = Eubacterium Inhibition of pathogenic

biogenic amines Metanogenic bacteria
Transformation de anaer. G(+) cocci Immune system
Bile salts stimulation
Sulfate reducers
8 Lactobacillus
Diarrhea E. coli Help in digestion and
Constipation
enterobacteria absorption of nutrients
Infections
and minerals
Systemic effects
Veillonella
Vitamin Synthesis
Clostridium
Staphylococcus
Salmonella Vibrionaceae
80% unknown/unclassified
Ps. aeruginosa Only
2
1 to 5% cultivable
G.R Gibson, 1998 ( )
Populations, log cfu/g fezes
Healthy Guts May Help to Reduce
Gas Emissions, Odors and
Environmental Impact
Less Ammonia, Less Animal Welfare
and Processing Issues
 Outline of this Presentation
● Importance of intestinal health
● Gut microbiome and its impacts
● Factors that disrupt gut ecosystem affecting health
 Factors that Disrupt Gut Ecosystem Equilibrium
● Stress: temperature,
• Toxins: mycotoxins,
environmental factors (light)
biogenic amines, rancid fat
● Water: excess salts, organic
matter, pH, temperature
• Parasites: coccidia,
worms, histomonas, hexamita
● Litter: ammonia, air quality
• Virus: entero, reo, corona,
• Diet: particle size, form
astro, rota, adeno, alphi,
byproducts, fiber Newcastle
● Antinutrients: trypsin inhibitors,
• Bacteria: Campylobacter,
phytate, NSPs, ovecooked
Clostridium, Salmonella, E. coli
protein, AAimbalance, fat factors,
calcium levels
 Intestinal Barriers to Gut Pathogen
Translocation
1. Physical barriers 5. Polymeric
2. Peristalsis (transit time) Immunoglobulins
3. Secretions (water, 6. Physical properties,
electrolytes, HCl, pH in each section
enzymes, bile, Ig A)
7. Mucus - mucins
4. Continuous mucosa cell
sloughing 8. Associated flora
 2. Normal Gut Motility in Avian Species
• Peristaltic refluxes
• Passage rate could be altered
by nutrient density, diet
digestibility, and osmotic
properties
Adequate particle size
The stimulates gizzard action
Gizzard Normalizes feed passage rate
“Pace-setter” of Gut
Motility
Reverse Peristalsis • Gastric reflux:
Gizzard to Proventriculus

•Upper intestinal reflux

Jejunum to Gizzard

• Lower intestinal reflux

Colon to Ceca 27
Gut Motility Affects Enteric Ecosystem
• Vigorous gut refluxes or antiperistalsis
compensates for a short intestine in birds
• Digesta exposure to gastric secretions
• Vigorously mix digesta with enzymes
• Enhances nutrient absorption
• Discourages microbial proliferation
• Fine cereal grinding and extensive feed
processing contribute to gut health problems
Optimal Rate of Digestion Results in Little
Substrate for Bacteria
Absorbed nutrients

Starch
Fat
Protein

Fewer Bacteria Bedford (2002)

 Slow digestion by the bird leads to more
substrate for bacteria
Fewer absorbed nutrients

Starch
Fat

Protein

More Bacteria
Bedford (2002)
Feed Passage
 Slow digestion by the bird leads to more
substrate for bacteria
Fewer absorbed nutrients

Starch
Fat
Acute
Protein response
dysbacteriosis
or enteritis
More Bacteria
Bedford (2002)
 Slow digestion by the bird leads to more
substrate for bacteria
Fewer absorbed nutrients

Starch
Fat
Acute
Protein response
dysbacteriosis
or enteritis
Chronic More Bacteria Bedford (2002)
Response is to produce more enzymes, immunological reaction and
grow a larger intestine. Costly in nutrient - energy terms.
Proventriculitis and
gizzard atrophy
 6. Physical properties, pH
pH 2,5 pH 3,5
pH 6,5 -7,0

pH 7,0

pH 7-8

pH 5,0
 Feeding Mash Particle Size (Exp. 1) and Pelleting a Fine
Mash on the Relative Weights of Gizzard and Small
Intestine and the pH of their Contents
Gut parameters Experiment 1 Experiment 2
Mash Texture Fine Mash P<
Fine Medium Coarse Mash Pellet
Relative Weight
(g/100 g BW)

Gizzard 3.95 5.50 5.97 2.92 1.82 0.01

Small Intestine 6.61 5.71 5.98 4.95 5.68 0.05
 Feeding Mash Particle Size (Exp. 1) and Pelleting a Fine
Mash on the Relative Weights of Gizzard and Small
Intestine and the pH of their Contents
Gut parameters Experiment 1 Experiment 2
Mash Texture Fine Mash P<
Fine Medium Coarse Mash Pellet
Relative Weight
(g/100 g BW)

Gizzard 3.95b 5.50a 5.97a 2.92a 1.82b 0.01

Small Intestine 6.61a 5.71ab 5.98b 4.95b 5.68a 0.05
pH of contents
Gizzard 3.47a 3.03a 2.74b 3.29 3.45 0.05
Small Intestine 5.92b 6.26a 6.32a 5.98 5.49 0.05
 6. Physical properties, pH
pH 2,5 pH > 3,5
pH <6,5 -7,0

pH < 7,0

pH < 7-8

pH < 5,0
 Symptoms of Poor Gut Motility
• Proventriculitis and gizzard atrophy
• Increased feather and litter picking
• Poor protein and fat digestion
• Reduced feed conversion, Increased Feed
passage
• Increased susceptibility to colonization of enteric
pathogens
• Poor water and electrolyte reabsorption
• Increased mortality rate
 Outline of this Presentation
● Importance of intestinal health
● Gut microbiome and its impacts
● Factors that disrupt gut ecosystem affecting health
● Current methods to study microbial ecology
 Methods to Study Intestinal Microflora
• Traditional Methods based on:
• Anaerobic culture techniques
• Phenotypic characterization
• Enumeration in selective media
• Optic or electronic microscopy

• Only 1% of gut microbial communities are cultivable

and 8% known and named (Hugenholtz et al., 1998).

• Culturemethods can not simulate gut environment

conditions, especially ecological relations.
• Data from culture methods are very variable and
always incomplete.
Gut microbial ecology is much bigger than expected
No gut microbial community acts by itself
 Differences?
It’s the same microflora?
Gel Eletroforeses
Pyrosequencing
 Krona chart of bacteria
from chicken cecum (from
919 of the 972 chicken cecal sequences)
Low Relative Diversity

Most
predominant
Clostridium,
Ruminococccus,
Lactobacillus,
and Bacteroids
Wei et al., 2013. PS 92: 671-683
Distribution of bacterial
genera identified in the
cecal microbiomes of
chicken and turkey.
Red bars: chicken cecum;
green bars: turkey cecum.

Chickens and
turkeys only share
16% similarity at
species level
Wei et al., 2013. PS 92: 671-683
 Cecal microbial communities from male broilers with
improved performance (▾) or poorer-performing birds (▴)

Torok V A et al. Appl. Environ. Microbiol. 2011;77:5868-5878

 Outline of this Presentation
● Importance of intestinal health
● Gut microbiome and its impacts
● Factors that disrupt gut ecosystem affecting health
● Current methods to study microbial ecology
● Alternatives to growth promoting antibiotics
 Alternatives to Growth Promotant
Antibiotics and Coccidiostats
Minerals Prebiotics
• Zinc Oxide • MOS - OS
• Copper Sulfate • FOS - Inulin
• Arsenicals Plant extracts
Exogenous enzymes • Saponins
• Carvacrol, timol,
• Proteases, Amilases
•Feed Processing and Composition
• ß-mannanase
• Xylanase, ß-glucans •Management practices and
Biosecurity
Organic acids
• Propionic, Acetic, Butiric
•Vaccination against specific pathogens
•Immuno-modulation
Probioticos
• Water or Spray Application •Genetic Selection
• Feed additive
Influence of probiotics and
antibiotics on goblet cells (Uni et al., 2005)

50 µm 50 µm 50 µm
Control Probiotic Antibiotic
 Effects of Essential Oils in the
Avian Gastro Intestinal Tract
ƒ Intestinal villus development 9

ƒ Mucin production 9
ƒ Enterocyte migration 8
ƒ Intestinal pH ~8

ƒ Pancreas estimulation ~9
ƒ Enzymes
 Effects of essential oil blend on ileal
mucin production in broilers at 46 d
300 a
Mucin Productrion (µg/g)

250
ab b
200
150
100
50
0
0 150 300
Essential oil + Benzoic acid concentration (ppm)
 Experience
with Eubiotic
Additives

Probiotics and
Essential Oils
 Experiment 1: Treatments
1. Positive Control (PC) antibiotic (BMD®) &
ionophore (Coban®)
2. Negative Control (NC) ionophore only
3. Probiotic 1 (B. coagulans) BC-30
4. Probiotic 2 (B. licheniformis, B. subtilis) B2B
5. Probiotic 3 (B. subtilis) Calsporin
6. Essential oil blend 1 (CPP) 300 ppm
7. Essential oil blend 1 (CPP) 150 ppm
12 replicates per treatment
* All diets contained ionophore up to 35 d
 Experiment 1
Feeding
Phase
Variable Mean SEM CV % P -value
BWG, kg 0.540 0.005 3.4 NS
Starter
FI, kg 0.800 0.006 2.9 NS
1 – 18 d
FCR, kg:kg 1.481 0.011 2.5 0.011
BWG, kg 1.379 0.009 2.4 0.130
Grower
FI, kg 2.295 0.016 2.5 NS
18 – 35 d
FCR, kg:kg 1.661 0.008 1.7 0.006
BWG, kg 0.700 0.009 4.8 0.033
Finisher
FI, kg 2.295 0.016 2.5 NS
35 – 43 d
FCR, kg:kg 1.965 0.021 3.6 0.025
Exp. 1 Starter 1-18 d

P = 0.79

P = 0.01
 Outline of this Seminar
• Intestinal Health
• Methods to Evaluate Microbiota
• Essential Oils and Probiotics
– Performance
– Microbial Ecology
 Lactobacilos, Lactobacilos,
Streptococos & Streptococos &
Coliformes Coliformes

Bifidobactérias,
Bacteroides,
Streptococos &
Clostridium

Effects on Gut Microflora?

 Evaluation of
Antibiotic Growth Promotant,
Ionophore,
Three Probiotics,
Two Essential Oils
Cecal contents at 43 days in
Two Consecutive Trials – Same Pens
Dendogram of Cecal Microbial Communities
in Broilers at 43 days Experiment 1

GPA + Ionophore
Ionophore

EO1 150ppm
Probio1

Probio2
EO1 300ppm
Probio3
Dendogram of Cecal Microbial Communities
in Broilers at 43 days Experiment 2

GPA + Ionophore
Ionophore
Probio1
Probio2
Probio3
EO1 300ppm

EO2 100ppm
 Summary Two Experiments
• Essential Oil Blend 1 (CPP) at 300 ppm
and probiotic 3 (B. subtilis) may be
consider as growth promotants.
• Each additive modulates gut microflora in
a different manner
 Effects of Essential Oils on
Corn and Wheat Diets
85 90 95 100

Similarity Coefficients
Corn,
A EO
46d 0ppm
86.5
Wheat, EO 0ppm
D 46d
80.3 Corn,BEO
46d150ppm
94.7
Corn,CEO
46d300ppm
86.6
Wheat,EEO
46d 150ppm
96.0
Wheat,FEO
46d300ppm
Additives: Antibiotics+ Ionophores, essential oils in
Coccidia vaccination and infection
• Corn + Soybean diets
• Additives:
BMD®+Coban®, Crina Poultry®, Crina Alternate®
• Gavage infection at 17 d:
E. acervulina, E. maxima, E. tenella
• Cecal samples collected 7 d post-infection
 Gut Microbial Ecology of Broilers Evaluated by DGGE and
Compared as SimilarityCoefficients (%SC)
Similarity Coefficients (%) between MC in Pre- and Post-Challenge
Periods
90 86.4
81.8 81.9
78.4 Duodenal Ileal Cecal 79.5

75
73.3
Similarity Coefficient

66.7 66.5 67.9

66.4
59.7 60.4
60
55.4

45

36.7 36.2

30
Unmedicated Unmedicated Infected Crina Poultry Crina Alternate
BMD + Coban
Uninfected Specific Essential Oils Blends

(Hume et al, 2006)

Pyrosequencing
Cecal Microflora in Broilers Fed
Basal Diet, Pre-infection 17 d

Vankley et al., 2013

 Cecal Microflora in Broilers Fed
Control - Basal Diet, Uninfected 26 d
Eimeria infection can cause severe changes on cecal microflora
Cecal Microflora in Broilers Fed
Basal diet, post- infection 26 d

Vankley et al., 2013

GPA (BMD) caused a partial modulation
post Eimeria-infection (26 d)

Vankley et al., 2013

Cecal Microflora of Broilers Fed Diet Supplemented with
EO Crina® Poultry, post Eimeria-infection (26 d)
Cecal Microflora of Broilers Fed Diet Supplemented with
EO Crina® Alternate, post Eimeria-infection (26 d)

Vankley et al., 2013

 Conclusions
• Intestinal health is key for live performance,
food safety, animal welfare and reduce
environmental impact of poultry production
• Intestines should be managed as ecosystems
• Understand gut microbial dynamics is
important to develop new additives, reduce
patogens and improve poultry performance
• Molecular methods are fundamental tools to
study gut Microbial Ecology
 Conclusions
• Each additive affects microflora in a different
manner
• Succesful and Sustentable Additives should
contribute to mantain microflora diversity
• Some additives may also affect the host
directly, not only the microbial communities
• Effects at host level should be understood and
used to improve holistic efficiency
Prestage Department of Poultry Science

International Course on Poultry Production, May 12-16, 2014

Feed Manufacturing Short-Course, May 16-17, 2014