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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 toGut 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, Streptococos &
Coliformes
Lactobacilos, Streptococos &
Coliformes
Bifidobactérias, Bacteroides,
Streptococos & Clostridium
Gut: Ecosystem with bird as host
Gut Ecosystem in Equilibrium = Healthy
Toxin production, potential carcinogens = AGV, Amonia
Intestinal putrefaction = biogenic amines
Transformation de Bile salts
Effects of Microbial Populations in the Intestine
Negative Positive
(G.R Gibson, 1998)
Bifidobacterium
Bacteroides
Eubacterium
Metanogenic
anaer. G(+) cocci
Sulfate reducers
Lactobacillus
enterobacteria
Veillonella
Clostridium
Staphylococcus
Vibrionaceae
Ps. aeruginosa
11
2
8
Populations, log cfu/g fezes
E. coliDiarrheaConstipationInfectionsSystemic effects
Inhibition of pathogenic bacteria
Immune system stimulation
Help in digestion and absorption of nutrients and minerals
Vitamin Synthesis
Salmonella
Edgar Oviedo
Pathobionts
Toxin production, potential carcinogens = AGV, Amonia
Intestinal putrefaction = biogenic amines
Transformation de Bile salts
Microbial Populations in the Intestine
Negative Positive
(G.R Gibson, 1998)
Bifidobacterium
Bacteroides
Eubacterium
Metanogenic
anaer. G(+) cocci
Sulfate reducers
Lactobacillus
enterobacteria
Veillonella
Clostridium
Staphylococcus
Vibrionaceae
Ps. aeruginosa
11
2
8
Populations, log cfu/g fezes
E. coliDiarrheaConstipationInfectionsSystemic effects
Inhibition of pathogenic bacteria
Immune system stimulation
Help in digestion and absorption of nutrients and minerals
Vitamin Synthesis
Salmonella
Edgar Oviedo
80% unknown/unclassifiedOnly 1 to 5% cultivable
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
• Toxins: mycotoxins, biogenic amines, rancid fat
• Parasites: coccidia, worms, histomonas, hexamita
• Virus: entero, reo, corona, astro, rota, adeno, alphi, Newcastle
• Bacteria: Campylobacter, Clostridium, Salmonella, E. coli
● Stress: temperature, environmental factors (light)
● Water: excess salts, organic matter, pH, temperature
● Litter: ammonia, air quality• Diet: particle size, form
byproducts, fiber●Antinutrients: trypsin inhibitors,
phytate, NSPs, ovecookedprotein, AAimbalance, fat factors, calcium levels
Intestinal Barriers to Gut Pathogen Translocation
1. Physical barriers
2. Peristalsis (transit time)
3. Secretions (water, electrolytes, HCl, enzymes, bile, Ig A)
4. Continuous mucosa cell sloughing
5. Polymeric
Immunoglobulins
6. Physical properties,
pH in each section
7. Mucus - mucins
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 stimulates gizzard action
Normalizes feed passage rateThe
Gizzard
“Pace-setter” of Gut Motility
27
•Upper intestinal refluxJejunum to Gizzard
• Lower intestinal refluxColon to Ceca
Reverse Peristalsis • Gastric reflux:Gizzard to Proventriculus
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
Starch
Fat
Protein
Fewer Bacteria
Absorbed nutrients
Bedford (2002)
Slow digestion by the bird leads to more substrate for bacteria
Starch
Fat
Protein
More Bacteria
Fewer absorbed nutrients
Bedford (2002)
Feed Passage
Slow digestion by the bird leads to more substrate for bacteria
Starch
Fat
Protein
More Bacteria
Fewer absorbed nutrients
Bedford (2002)
Acute response
dysbacteriosis or enteritis
Slow digestion by the bird leads to more substrate for bacteria
Starch
Fat
Protein
More Bacteria
Fewer absorbed nutrients
Response is to produce more enzymes, immunological reaction and grow a larger intestine. Costly in nutrient - energy terms.
Bedford (2002)Chronic
Acute response
dysbacteriosis or enteritis
Proventriculitis and
gizzard atrophy
pH 2,5 pH 3,5
pH 5,0
pH 6,5 -7,0
pH 7-8
pH 7,0
6. Physical properties, pH
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
P <Mash Texture Fine Mash
Fine Medium Coarse Mash PelletRelative Weight
(g/100 g BW)
Gizzard 3.95 5.50 5.97 2.92 1.82 0.01Small 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
P <Mash Texture Fine Mash
Fine Medium Coarse Mash PelletRelative Weight
(g/100 g BW)
Gizzard 3.95b 5.50a 5.97a 2.92a 1.82b 0.01Small Intestine 6.61a 5.71ab 5.98b 4.95b 5.68a 0.05pH of contents
Gizzard 3.47a 3.03a 2.74b 3.29 3.45 0.05Small Intestine 5.92b 6.26a 6.32a 5.98 5.49 0.05
pH 2,5 pH > 3,5
pH < 5,0
pH <6,5 -7,0
pH < 7-8
pH < 7,0
6. Physical properties, pH
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 cultivableand 8% known and named (Hugenholtz et al., 1998).
• Culture methods can not simulate gut environmentconditions, especially ecological relations.
• Data from culture methods are very variable andalways incomplete.
Gut microbial ecology is much bigger than expectedNo 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
Wei et al., 2013. PS 92: 671-683
Most predominant Clostridium,
Ruminococccus, Lactobacillus,
and Bacteroids
Distribution of bacterial genera identified in the
cecal microbiomes ofchicken 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• Zinc Oxide• Copper Sulfate• Arsenicals
Exogenous enzymes• Proteases, Amilases• ß-mannanase• Xylanase, ß-glucans
Organic acids • Propionic, Acetic, Butiric
Probioticos• Water or Spray Application• Feed additive
Prebiotics• MOS - OS• FOS - Inulin
Plant extracts• Saponins• Carvacrol, timol,
•Feed Processing and Composition
•Management practices and Biosecurity
•Vaccination against specific pathogens
•Immuno-modulation
•Genetic Selection
AntibioticProbioticControl50 µm 50 µm 50 µm
Influence of probiotics and antibiotics on goblet cells (Uni et al., 2005)
Effects of Essential Oils in theAvian Gastro Intestinal Tract
Intestinal villus development
Mucin production
Enterocyte migration
Intestinal pH ~
Pancreas estimulation ~Enzymes
Effects of essential oil blend on ileal mucin production in broilers at 46 d
a
ab b
0
50
100
150
200
250
300
0 150 300Mu
cin
Pro
du
ctri
on (
µg/
g)
Essential oil + Benzoic acid concentration (ppm)
Experience with Eubiotic
Additives
Probiotics and Essential Oils
Experiment 1: Treatments1. 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 1Feeding Phase
Variable Mean SEM CV % P -value
Starter
1 – 18 d
BWG, kg 0.540 0.005 3.4 NSFI, kg 0.800 0.006 2.9 NSFCR, kg:kg 1.481 0.011 2.5 0.011
Grower
18 – 35 d
BWG, kg 1.379 0.009 2.4 0.130FI, kg 2.295 0.016 2.5 NSFCR, kg:kg 1.661 0.008 1.7 0.006
Finisher
35 – 43 d
BWG, kg 0.700 0.009 4.8 0.033
FI, kg 2.295 0.016 2.5 NS
FCR, kg:kg 1.965 0.021 3.6 0.025
P = 0.79
P = 0.01
Exp. 1 Starter 1-18 d
Outline of this Seminar
• Intestinal Health
• Methods to Evaluate Microbiota
• Essential Oils and Probiotics
– Performance
– Microbial Ecology
Lactobacilos, Streptococos &
Coliformes
Lactobacilos, Streptococos &
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
Probio1
Probio2
Probio3
EO1 300ppm
EO1 150ppm
GPA + Ionophore
Ionophore
Dendogram of Cecal Microbial Communities in Broilers at 43 days Experiment 1
GPA + Ionophore
Ionophore
Probio1
Probio2
Probio3
EO1 300ppm
EO2 100ppm
Dendogram of Cecal Microbial Communities in Broilers at 43 days Experiment 2
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
A 46dD 46dB 46dC 46dE 46dF 46d
100959085
80.3
86.6
94.7
86.5
96.0
Corn, EO 0ppm
Wheat, EO 0ppm
Corn, EO 150ppm
Corn, EO 300ppm
Wheat, EO 150ppmWheat, EO 300ppm
Effects of Essential Oils onCorn and Wheat Diets
Similarity Coefficients
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
78.4
36.7
73.3
59.7
66.7
55.4
81.8 81.986.4
79.5
67.9
60.466.4
66.5
36.2
30
45
60
75
90
Sim
ilarit
y C
oeff
icie
nt
Duodenal Ileal Cecal
Unmedicated Uninfected
BMD + CobanUnmedicated Infected Crina Poultry Crina Alternate
Specific Essential Oils Blends
(Hume et al, 2006)
Pyrosequencing
Cecal Microflora in Broilers FedBasal Diet, Pre-infection 17 d
Vankley et al., 2013
Cecal Microflora in Broilers FedControl - Basal Diet, Uninfected 26 d
Cecal Microflora in Broilers FedBasal diet, post- infection 26 d
Eimeria infection can cause severe changes on cecal microflora
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, 2014Feed Manufacturing Short-Course, May 16-17, 2014
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