©2019, Elanco Animal Health
Efficacy of dried egg product administered to male broiler chickens during experimental necrotic enteritis
J. Escobar1, G. Mathis2, B. Lumpkins2, J. Sand3, N. Olmeda-Geniec1
1. Elanco Animal Health, 2. Southern Poultry Research
3. Ab E Discovery
18 December 2019
USDA 3rd International Symposium on Alternatives to Antibiotics
Bangkok, Thailand
©2019, Elanco Animal Health
Necrotic enteritis (NE) • “NE is the most common and financially devastating
bacterial disease in modern broiler flocks” (poultryhub.org)
• Wet litter, diarrhea, increase in mortality
• Depression of growth rate and feed efficiency
• Multifactorial disease
– Coccidiosis: the trigger!
– Clostridium perfringens
• Current efforts:
– Vaccination, medication,
and management 2 Source: poultryhub.org
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Interleukin-10 IL-10 is a potent anti-inflammatory cytokine that decreases:
• Innate immune response
– MHC Class II proteins, costimulatory molecules, and reactive oxygen
intermediates
• Adaptive immune response
– Antigen specific T-cell generation
• Inflammation
– Inhibits production of IFN-γ, TNF-a, IL-1, IL-2, IL-6 and GMCSF
IL-10 main function
• “Resolve” inflammatory processes
3
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IL-10 reduces IFN-g production and release in ConA-stimulated chicken splenocytes in vitro
4 Source: Arendt et al., 2016. Poult Sci 95:430
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Role of IL-10 in pathogenesis
• Epstein Barr Virus encodes for IL-10 and inhibits anti-viral response
• Yersinia pestis upregulates IL-10 production and favors its survival
• Leishmania donovani upregulates IL-10 and IL-10 KO mice are resistant
• Eimeria resistant chickens express low IL-10
• Eimeria infection increases IL-10 levels
5 …. a cloaking mechanism
Certain pathogens appear to use high expression of IL-10 to thwart
and fool an immune response Source: hindustantimes.com
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Hypothesis
6
Neutralization of intestinal secreted IL-10 with an IgY may contribute to increase IFN-g production to elicit an appropriate immune response resulting in better growth performance of broilers with necrotic enteritis
VL VH
CL
Cv3
Cv4
Cv2
Cv1
©2019, Elanco Animal Health
Dried Egg Product (DEP)
• Patents: 8652457; US20150037277; 9505836; US20160280778; US20160008436
• Laying hens are hyper-immunized with a patented
peptide against chicken IL-10 (Arendt et al., 2016 Poult. Sci. 95:430)
• Eggs containing anti-chicken IL-10 IgY antibodies
• Eggs are dried in a manner to
preserve IgY binding activity
7
VL VH
CL
Cv3
Cv4
Cv2
Cv1
©2019, Elanco Animal Health
Experimental design and analyses
• Randomized complete block design
• Six DEP doses (0, 143, 287, 358, 430, and 573 U/MT)
– Starter and Grower; no Finisher
• Cobb 500 males
• 50 birds/pen
• 70 floor pens (10 pens/treatment)
• Study length: 42 d
– Starter (d 0 to 14), Grower (d 14 to 28), and Finisher (d 28 to 42)
• Southern Poultry Research
8
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Experimental design and analyses, continued
• Experimental necrotic enteritis
– Coccivac® B52 (Merck Animal Health, Kenilworth, NJ) spray vaccination on day of hatch according to label
– Clostridium perfringens
• 108 cfu/bird/d in feed
• Day 18, 19 and 20 of study
• 4 bird/pen randomly selected for NE lesion scoring on d 20
• Growth performance
– Each feeding phase and overall
• Pen was the experimental unit with treatment as a fixed effect and block as a random effect (JMP v.14.1)
9
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Results
10
Source: wikipedia.org
©2019, Elanco Animal Health ©2019, Elanco Animal Health
DEP IgY, U/MT0 143 287 358 430 573
NE
le
sio
n s
co
re (
d 2
0)
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
No
n-i
nfe
cte
d
a
a,b
b,c
b,c
b,cc
0%
-19%
-28%
-38%
-44%-47%
a,b,c,d P<0.05
Infe
cte
d
Infe
cte
d
Infe
cte
d
Infe
cte
d
Infe
cte
d
Infe
cte
d
d
-100%
Necrotic enteritis lesion scores (d 20)
11
©2019, Elanco Animal Health
Dried egg product IgY, U/MT
0 143 287 358 430 573
Ov
era
ll A
DF
I (d
0-4
2),
g/d
0
10
20
30
40
50
60
70
80
90
100
110
No
n-i
nfe
cte
d
a
c
a,b a,b a,ba,bb
a,b,c P<0.050%
-7%
-1% -1% -1%-3%
0%
Infe
cte
d
Infe
cte
d
Infe
cte
d
Infe
cte
d
Infe
cte
d
Infe
cte
d
Overall average daily feed intake (d 0-42)
12
Linear P<0.79
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Overall average daily gain (d 0-42)
13
Dried egg product IgY, U/MT
0 143 287 358 430 573
Ov
era
ll A
DG
(d
0-4
2),
g/d
0
55
56
57
58
59
60
61
62
63
64
No
n-i
nfe
cte
d
a
b
b
b
b,c
d
c,d
0%
-2%
+4%
+3%
+5% +5%
+8%
a,b,c,d P<0.05
Infe
cte
d
Infe
cte
d
Infe
cte
d
Infe
cte
d
Infe
cte
d
Infe
cte
d
©2019, Elanco Animal Health
DEP IgY, U/MT0 143 287 358 430 573
Ov
era
ll F
CR
(d
0-4
2)
0.00
1.66
1.68
1.70
1.72
1.74
1.76
1.78
1.80
1.82
1.84
1.86
No
n-i
nfe
cte
d
a
b bb
b,c
dc,d
0 pts
-9 pts -9 pts-8 pts
-10 pts
-13 pts-14 pts
a,b,c,d P<0.05
Infe
cte
d
Infe
cte
d
Infe
cte
d
Infe
cte
d
Infe
cte
d
Infe
cte
d
Overall feed conversion ratio (d 0-42)
14
©2019, Elanco Animal Health
Dried egg product IgY, U/MT
0 143 287 358 430 573
Ov
era
ll P
EI (d
0-4
2)
0
270
280
290
300
310
320
330
340
350
No
n-i
nfe
cte
d
c
c
bb
a,b
a
a,b
0%
+4%
+11%+12%
+14%
+16%
+19% a,b,c,d P<0.05
Infe
cte
d
Infe
cte
d
Infe
cte
d
Infe
cte
d
Infe
cte
d
Infe
cte
d
Overall production efficiency index (d 0-42)
15
PEI = ADG*Survival FCR*10
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Conclusion • Use of dried egg product containing anti-chicken-IL-10 may be a
viable alternative to reduce the negative effects of coccidiosis and
thus ameliorating necrotic enteritis in broilers
16
Summary
• Inclusion of dried egg product containing anti-chicken-IL-10:
– Reduced necrotic enteritis lesions
– Improved growth performance
©2019, Elanco Animal Health
References
• Arendt et al. 2016. Interleukin-10 neutralizing antibody for detection of intestinal luminal
levels and as a dietary additive in Eimeria challenged broiler chicks. Poult. Sci. 95:430-438.
• Ohga et al. 2002. Immunological aspects of Epstein-Barr virus infection. Crit. Rev.
Ocol/Hematolog 44:203-215.
• Ping Ouyang et al. 2014. IL-10 encoded by viruses: a remarkable example of independent
acquisition of a cellular gene by viruses and its subsequent evolution in the viral genome. J.
Gen. Virology 95:245–262
• Rothwell et al. 2004. Cloning and characterization of chicken IL-10 and its role in the immune
response to Eimeria maxima. J. Immunol. 173:2675-2682.
• Yu et al. 2008. Measles virus infection in adults induces production of IL-10 and is associated
with increased CD4+CD25+ regulatory T Cells. J. Immunol. 181:7356-7366.
• Redpath et al. 2014. Protection and pathology during parasite infection: IL-10 strikes the
balance. Parasite Immunol. 36:233-252.
17
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Thank you, questions?
18
VL VH
CL
Cv3
Cv4
Cv2
Cv1
International Poultry Scientific Forum at IPPE 2020
• Efficacy of dried egg product administered to chickens during an enteritis infection caused by Eimeria spp. and Clostridium perfringens
• Binding kinetics of IgY contained in a dried egg product to recombinant chicken IL-10
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