Phytonutrients 101 - Presented by: Yvette La-Garde, Director of Education of VitaMedica
Dietary phytonutrients enhance disease resistance … phytonutrients enhance disease resistance of...
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Dietary phytonutrients enhance disease resistance of pigs
Yanhong Liu,*,1 D. Bravo,2 and J. E. Pettigrew3
University of California, Davis, USA1, Pancosma SA, Geneva, Switzerland2, and University of Illinois, Urbana, USA3
UCDAVIS
2nd International Symposium - Alternatives to Antibiotics, 2016
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Outline
• Previous knowledge
• Hypothesis
• Test of hypothesis
• What does it mean?
UCDAVIS
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Weaning stress
• Maternal separation
• Environmental change
• Increased exposure to pathogens
• Social hierarchy stress
• Move to solid feed
• Transportation stress
UCDAVIS
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Gut morphology change of weanling pigs
• Reduced feed intake
• Negative effects on intestinal morphology
UCDAVIS
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Declined intestinal functions
• Reduced brush-border enzyme activity
• Reduced absorption ability
UCDAVIS
• Diarrhea
• Poor growth performance
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Immunity of weanling pigs• Passive immunity is declining• Active immunity is not fully developed
UCDAVIS
• Highly sensitive to infectious disease
• Divert nutrients away from growth to immune response
• Poor growth performance
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Antibiotic use on farms
UCDAVIS
1
2
3
Van Boeckel et al., 2015 PNAS
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Feed additives
UCDAVIS
• Mannan oligosaccharides
• Immune egg products
• Direct-fed bacteria
• Yeast/yeast products
• Plant extracts
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Plant extracts
UCDAVIS
• Extracted from parts of plants or synthesized
• Concentrated, hydrophobic, volatile aroma
• Mixtures of secondary plant metabolites
• Liquid or powder
• Phenolic compounds
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Plant extracts
UCDAVIS
• Biological effects:
Antimicrobial Anti-inflammatoryAntioxidantOthers: Antiviral, Antifungal, Antiparasitic,
Antitoxigenic
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Hypothesis
UCDAVIS
1) Certain plant extracts modify immune function of pigs
2) This leads to increased disease resistance
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Test of hypothesis
UCDAVIS
• Exp. 1: In vitro cell culture
• Exp. 2: E. coli challenge study
• Exp. 3: PRRS challenge study
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Experiment 1
UCDAVIS
In vitro cell culture assays
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Anti-inflammatory effects
UCDAVIS
Liu et al., 2012
**
*
*
*
**
*
** *
*
*
*
*
** *
*
*
* * *0
10
20
30
40
50
ng/m
L
02550100200
TNF-α *P < 0.05
LPS-stimulated porcine alveolar macrophages
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Anti-inflammatory effects
UCDAVIS
Liu et al., 2012
*
*
*
**
*
*
**
**
*
** **
*
0
2
4
6
8
10
12
14
ng/m
L
02550100200
IL-1β *P < 0.05
LPS-stimulated porcine alveolar macrophages
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Conclusions – Exp. 1
UCDAVIS
• All of plant extracts used in this experiment may have potent anti-inflammatory effects
• Carvacrol, cinnamaldehyde, eugenol, and garlicon might be the more powerful candidates
• Capsicum oleoresin, garlicon, and turmeric oleoresin were selected to do E. coli and PRRSV challenge studies
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In vivo E. coli challenge study
UCDAVIS
Liu et al., 2013a
Experiment 2
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UCDAVIS
Liu et al., 2013a
Procedures
d-4
d0
d3
Weaning (21 d old)
Daily diarrheaFecal culture
F18 E. coli inoculation (d0 to d2)
Room
ChamberWhite blood cell countsCytokinesAcute phase proteins
Sows screening
ImmunohistochemistryIntestinal morphologyqPCR & Microarray (d 5)
* 4 diets: control, 10 ppm capsicum oleoresin, 10 ppm garlicon, 10 ppm turmeric oleoresin
*
d5
d8
d11
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UCDAVIS
05
1015202530354045
d0-11 PI d0-11 PI
Sham E. coli
ControlCapsicumGarliconTurmeric
%
Control vs. plant extractsP < 0.05
Liu et al., 2013a
Control vs. plant extractsP < 0.05
Pig days with diarrhea score ≥ 31, normal; 5, watery diarrhea
Frequency of diarrhea
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Liu et al., 2013a, 2014
Possible mechanism for reduced diarrhea
* * *
050
100150200250300350400450
Sham E. coli
μm
* *
0
30
60
90
120
150
180
E. coli
ControlCapsicumGarliconTurmeric
Possibly improved gut barrier function!
Ileal villi height (d 5 PI) MUC2 in Ileal mucosa (d 5 PI)
UCDAVIS
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020406080100
Control Capsicum Garlicon Turmeric
Plant extracts reduced systemic inflammation caused by E. coli infection
Liu et al., 2013a
*
*
*
* *
0
5
10
15
20
25
30
35
40
d5 PI d11 PI
×10
00/μ
L
White blood cell counts
* ** *
0
20
40
60
80
100
120
d5 PI d11 PI
pg/m
L
Serum TNF-α
UCDAVIS
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*** *
**
0
500
1000
1500
2000
2500
Macrophage Neutrophil
Control CapsicumGarlicon Turmeric
Plant extracts reduced gut inflammation caused by E. coli infection
Liu et al., 2013a
UCDAVIS
Ileal mucosa, d 5 PI
ShamMacrophage
E. coliMacrophage
Ileum (d 5 PI)
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Plant extracts reduced gut inflammation caused by E. coli infection
Liu et al., 2014
UCDAVIS
*** *
*
*
0
50
100
150
200
250
COX-2 TNFA
Control CapsicumGarlicon Turmeric
The Prostaglandin Pathway
Arachidonic acid
PGG2
PGH2
TXA2
PGF2αPGE2
PGD2PGI2
Inflammation!
E. coli challenge group
Cyclooxygenase-2 (COX-2)
TNF-α
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Conclusions – Exp. 2
UCDAVIS
• Feeding plant extracts reduced diarrhea and enhanced disease resistance of weanling pigs
• Possible mechanisms• Gut barrier function• Gut mucosa immunity • Systemic immunity
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In vivo porcine reproductive and respiratory syndrome virus (PRRSV)
challenge study
UCDAVIS
Liu et al., 2013b
Experiment 3
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Rectal temperature
UCDAVIS
Liu et al., 2013b
39.0
39.5
40.0
40.5
41.0
d 0 d 4 d 7 d 9 d 11 d 14
˚C
CONCAPGARTURCON+PRRSVCAP +PRRSVGAR+PRRSVTUR+PRRSV
d 7, 9, 11: PRRSV: P < 0.01
* CAP vs. CON: P < 0.05# GAR vs. CON: P < 0.05
* #
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Feed efficiency, d 0-14
UCDAVIS
Liu et al., 2013b
*
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
Sham PRRSV
ADG
:AD
FI
ControlCapsicumGarlicTurmeric
PRRSV: P = 0.07
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* *
*
10
13
16
19
22
25
28
31
d 7 d 14
Ct v
alue Control
CapsicumGarlicTurmeric
Serum viral load-PRRSV
UCDAVIS
Liu et al., 2013b
CON vs. PE: P < 0.05
d 7 & 14: PRRSV: P < 0.01
CON vs. PE: P = 0.05
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Serum TNF-α - PRRSV
UCDAVIS
Liu et al., 2013b
* *
0
50
100
150
200
250
d 7 d 14
pg/m
L ControlCapsicumGarlicTurmeric
CON vs. PE: P < 0.05
d 7 & 14: PRRSV: P < 0.01
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*
0
10
20
30
40
50
60
d 7 d 14
µg/m
L ControlCapsicumGarlicTurmeric
Serum C-reactive protein- PRRSV
UCDAVIS
Liu et al., 2013bd 7 & 14: PRRSV: P < 0.01
CON vs. PE: P < 0.05
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Conclusions – Exp. 3
UCDAVIS
• Feeding plant extracts delayed fever caused by PRRS infection
• Feeding plant extracts improved feed efficiency of pigs
• Possible mechanisms• Reduced viral load• Reduced systemic inflammation
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Hypothesis
UCDAVIS
1) Certain plant extracts modify immune function of pigs Accept
2) This leads to increased disease resistance Accept
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Acknowledgements
• Dr. Pettigrew lab – University of Illinois
• Pancosma
UCDAVIS
2nd International Symposium - Alternatives to Antibiotics, 2016
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UCDAVIS
2nd International Symposium - Alternatives to Antibiotics, 2016
Comparative Animal Nutrition & Physiology Laboratory
http://animalnutr-ansci.faculty.ucdavis.edu/