Science, trial data, solutions

Science,
trial data,
solutions
ORGANISED BY:
Supporting The Immune System Of Piglets During The First Weeks Of Life By Feeding Egg Immunoglobulins
Fellipe Barbosa
Product Manager Globigen® Product Line
Agenda:
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Background
Egg immunoglobulins and IgY concept
Basic Immunology
Experimental data
Take home messages
Background
• Young mammalians are protected against diseases by specific
antibodies:
• Passive immunity  mother through milk/colostrum
• Active immunity  produced by the organism after contact
with a pathogen
• Some cases  The maternal barrier between the fetus and the
womb of some animals impedes the transference of parental
immunity.
• Young animals are born without an own immune protection and
the supply of antibodies from colostrum (passive immunization)
shortly after birth is therefore of vital importance.
Background
•
Piglets are born without immune protection •
Essential supply of immunoglobulins by colostrum
•
The immunoglobulin level in sow colostrum rapidly decreases during the first 24 hours after birth
•
Transfer of immunoglobulins through the intestinal wall of the piglet decreases
Markowska-Daniel and Pomorska-Mol, 2010
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The second critical period is the time around weaning  growth, and
subsequent performance of the piglet.
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Non‐digested ingredients ends up as a substrate for the multiplication of
pathogenic bacteria.
•
Disturb the balance of the microflora by competitive exclusion of beneficial
microbes, causing the severe symptoms of diarrhea.
•
Different strains of ETEC  diarrhea in weaned piglets (Fairbrother et al.,
2005).
•
da Rosa et al. (2015), the fimbrial types F4 and F18 are related to diarrhea
in weaned piglets and the fimbriae from these pathogens will attach to the
microvilli surface initiating bacterial colonization (Zhang et al., 2007).
Pathogenic germs of suckling piglets
C. perfringens types A/C
Escherichia coli
Rotavirus
Cryptosporidium
TGE virus
Immunity gap in young animals
Egg immunoglobulins and the IgY concept
History
• 1893: Dr. Felix Klemperer: Published the first article showing that
hens produced antibodies against tetanus bacteria and transferred
them into the egg. The egg yolk could be used successfully in mice
• 1969: Hideo Tokoro: Concept “From anti‐bio to pro‐bio“ with the
aim to develop natural alternatives to feed antibiotics.
The IgY concept
Antibodies are formed
Transfer into the eggs
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Immunoglobulin = Antibody
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Ig = Abbreviation for Immunoglobulins
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Antigen = Pathogen
Production
Binding of Pathogens at the target site
• Basic Immunology
• How does it work?
Source: Wikipedia
Basic Immunology
• Ig are proteins used by the immune
system to identify and neutralize
foreign substances
FAB
• FAB – Fragments of Antigen‐Binding
• V – Variable regions:
• Antigen‐specific design
• They react in a key‐and‐lock model
Wikipedia.org\antibody (picture)
The IgY concept
• Stein: “If it is correct that it is the immunoglobulins in spray dried
plasma that provide the improvement in pig immunity then it would
be expected that other sources of immunoglobulins also can improve
the immune status of pigs.”
• It has been demonstrated that if eggs containing high
concentrations of antibodies against E.coli are fed to weanling pigs,
these pigs will experience fewer cases of E.coli associated diarrhea
(Yokoyama et al., 1997; Marquardt et al., 1999).
• Egg proteins also contain Ig and the inclusion of eggs to weaning diets
may also improve pig performance. If hens are challenged with
certain pathogens, they will produce antibodies linked to these
pathogens (Schmidt et al., 2003; Hong et al., 2004).
> Improvement of immune status in piglets.
Mode of Action
1) Adherence inhibition of pathogens
2) Neutralization of viruses 3) Agglutination
1) Mode of Action – Adherence inhibition
Mechanism of pathogenesis due to ETEC
water
water
Toxins
water
Epithelial cells
Enterotoxins release from colonizing ETEC strain and trigger severe diarrhea, thus causing considerable economic losses.
Mechanism of pathogenesis due to ETEC
IgY
Bacterial fimbriae
Villi
Epithelial cells
IgY can effectively block the initial stages of the colonization process
2) Mode of Action – Neutralization (viruses)
• IgY prevent invasion of viruses so they cannot enter the host cells
• Viruses are inactivated
Mechanism of pathogenesis due to viral infection
water
Virus
water
water
Villi
Epithelial cells
Virus replication results in intestinal villous epithelial cell dysfunction / death and digestive and absorptive functions rapidly decrease
IgY can effectively block initial stages of infection process
IgY
Virus
Villi
Epithelial cells
IgY can effectively block initial stages of the infection process
3) Mode of Action – Agglutination
• IgY “glue” antigens/foreign particles together and stop their
movement
• IgY help immune cells (macrophages) to catch and eliminate
pathogenic germs
• IgY indirectly enhance the immune status of the host (Erhard et al.
1999
Serum IgG concentration
Globigen
Control
days after birth
Calculated total serum IgG
Globigen
Control
days after birth
Experimental data
Passive immunization of IgY against K88‐, K99‐, and 987P‐fimbriae by experimental enterotoxigenic
Escherichia coli infection in neonatal piglets
Infection and Immunity (1992, 60: 998‐1007)
Egg Immunoglobulins against ETEC
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76 colostrum‐deprived newborn piglets
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Antibody: against K88, K99, and 987P fimbria of enterotoxigenic Escherichia coli (ETEC)
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Groups: Control and IgY (1 titer, 4 titers, 16 titers)
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Test period: 1 to 7 days of age
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Mortality, Clinical symptoms and Bacteria detection
Egg Immunoglobulins against ETEC
Influence on diarrhea
Incidence of diarrhea (%)
100
80
60
40
20
0
0
Control (n=7)
1
Days after challenge
1 (n=7)
3
4 (n=7)
5
16 (n=7)
Attachment inhibition (bacteria)
(Yokoyama et al. 1992)
E. coli challenge trial with piglets
Control IgY
Experimental data
Efficacy of hyperimmunized hen egg yolks in the control of diarrhea in newly weaned piglets
Daniele Pozzebon da Rosa, Maite de Moraes Vieira, Alexandre Mello Kessler, Tiane Martin de Moura, Ana Paula Guedes Frazzon, Concepta Margaret McManus, Fábio Ritter Marx, Raquel Melchior & Andrea Machado Leal Ribeiro (2015): Efficacy of hyperimmunized hen egg yolks in the control of diarrhea in newly weaned piglets, Food and Agricultural Immunology., 2015
DOI: 10.1080/09540105.2014.998639
Challenge Trial – Brazil (da Rosa, 2015) Trial facility:
Objective
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The main objective of this study
was to evaluate the efficacy of
hyperimmunized hen egg powder
in the diarrhea control in newly
weaned piglets challenged with
F4 and F18 ETEC strains.
Trial Duration:
Sex:
Treatments:
Federal University of Rio Grande do Sul, Brazil
28 days (4 d of adaptation and 24 d of trial)
Male
T1 PC immune stimulated ETEC F4
(K88) + F18
T2 Immune stimulated ETEC F4 (K88)
+ F18 + 2kg/t Globigen® Jump
Start
T3 Immune stimulated ETEC F4 (K88)
+ F18+ 40 kg/t Plasma
T4 Negative Control with AGP –
colistin and zinc oxide – NC
N˚ of animals: Age at start: Criteria:
32 weaned piglets
22 days (average weight 5.4 ± 0.4 kg) Body weight (BW)
Feed intake
Average weight gain (WG)
Feeding efficiency (FE)
Fecal consistency score
Results – Animal performance
1 to 7 days after adaptation period
T1
T2
T3
T4
BW (kg)
6.54a
7.64bc
6.80ab
7.94c
Feed intake (kg)
1.46a
2.16ab
1.93ab
2.47b
WG (kg)
0.78a
1.88bc
1.04ab
2.19c
FE
0.38a
0.80ab
0.45ab
0.89b
Means followed by letters a,b,c in the same group of columns differ (p < 0.05).
BW  1.10 kg higher than the challenged animals!
da Rosa, 2015
Results – Animal performance
1 to 24 days after adaptation period
T1
T2
T3
T4
BW (kg)
13.82a
15.73ab
15.15ab
16.39b
Feed intake (kg)
10.45a
12.82ab
13.08ab
13.61b
WG (kg)
8.06a
9.97ab
9.38ab
10.62b
FE
0.76ab
0.78b
0.71a
0.78b
Means followed by letters a,b,c in the same group of columns differ (p < 0.05).
BW  1.91 kg higher than the challenged animals!
FI  2.37 kg higher than the challenged animals!
da Rosa, 2015
Results
Fecal Consistency Score 3.0
2.5
Score
2.0
1.5
1.0
0.5
0.0
1
2
3
4
5
6
7
8
9
Days of observation after the first inoculation T1
Globigen®
Egg Powder
T3
T4 (AGP)
da Rosa, 2015
Conclusions
• “In conclusion, the study demonstrated that the use of a
product based on egg yolks from hyperimmunized hens
(GLOBIGEN®*) containing antibodies against E. coli ETEC
reduced the severity of diarrhea and improved performance
of challenged animals.”
• “The yolks from hyperimmunized hens are a source of
antibodies competing with the porcine plasma and antibiotics,
not only to fight post‐weaning diarrhea, but also to prevent it,
resulting in a reduction in economic loss during this phase.”
*GLOBIGEN® is the exclusive brand of EW Nutrition for egg powder based products.
Experimental data
Prophylactic use of Ig‐PRO® in piglets
National Research Institute of Poland, 2009
• Ig‐PRO® is the exclusive brand of EW Nutrition for veterinary distribution.
• Most important ingredient: egg immunoglobulins (IgY) in high concentration.
• Main Ingredients: Oil , Immunoglobulins, Vitamins, Probiotics
Trial: Ig‐PRO®P University trial, Poland, 2009
• Treatment group (35 piglets, 3 litters):
• 2 ml Ig‐PRO® P on 1st day of life
• 1 ml on 2nd and 3rd day of life
• Control group (32 piglets, 3 litters)
• No treatment with Ig‐PRO® P
• Routine prophylactic program (Iron days 3+14, Toltrazuril days 3+4)
• Parameters evaluated:
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Incidence of diarrhea
Average daily gain
Blood count and differential blood count
Immunophenotyping and quantitation of Lymphocytes via flow cytometry (CD3+, CD21+, CD4+. CD8+, CD4+CD8+)
Trial: Ig‐PRO®P University trial, Poland, 2009
Performance parameters
Group
Incidence of Diarrhea
Daily weight gain from
farrowing to weaning
[g] Mean ±SD
Experimental Group Ig‐Pro P
1,95 / Litter
209.71* ± 53
Control
2,94 / Litter 175.21* ± 34
Data Summary ‐ University trial, Poland, 2009
• Treatment with Ig‐PRO® P reduced incidence of diarrhoea.
• Higher Average daily gains in the treatment group (+ 34,5g).
• Transient increase in total Leucocytes until 7 days after
prophylactic application of Ig‐PRO® P.
• Transient increase in Monocyte count until 7 days after
prophylactic application of Ig‐PRO® P.
• Transient increase of CD4+ T‐helper cells in Ig‐PRO ® P group.
• Persistent increase of CD4+/CD8+
Lymphocytes in Ig‐PRO ® P group.
effector/memory
T‐
Conclusion ‐ University trial, Poland, 2009
• Through prophylactic use of Ig‐PRO® P the general health
status of piglets can be improved and stabilized.
• Treatment with Egg derived Immunoglobulins induces
stimulatory effects on the innate immune response as well as
on adaptive immunity.
Experimental data
Globigen® Pig Doser Trial
Nakornratsrima, Thailand
• Globigen® Pig Doser is a commercial brand of EW Nutrition.
• Most important ingredient: egg immunoglobulins (IgY) in high concentration.
• Main Ingredients: Oil , Immunoglobulins, Vitamins, Probiotics
Trial design
• Treatment group:
• Globigen® Pigdoser per oral continuous 2 days after birth at 1 dose (2ml)/day.
• Control group:
• normal management practice – No Pig Doser
• Parameters evaluated:
• % Livability
• Weight at weaning (kg/pig)
• Weight gain (kg/pig)
• ADG (g/day)
• % Cull and mortality
Results: Thailand
Globigen®
Control
% Improve
No. of piglet at beginning
188
192
No. of piglet at weaning
175
171
% Livability
93.1
89.1
Weight at beginning (kg/pig)
1.8
1.8
Weight at weaning (kg/pig)
6.9
6.5
7.0
Weight gain (kg/pig)
5.1
4.7
8.8
ADG (g/day)
284.9
261.8
8.8
% Cull and mortality
16.7
20.5
18.2
4.5
Experimental data
Globigen® Pig Doser Trial
Nakornpratom, Thailand
• Globigen® Pig Doser is a commercial brand of EW Nutrition.
• Most important ingredient: egg immunoglobulins (IgY) in high concentration.
• Main Ingredients: Oil , Immunoglobulins, Vitamins, Probiotics
Trial design
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Treatment group:
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Globigen® Pigdoser per oral continuous 2 days after birth at 1 dose (2ml)/day.
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Control group:
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normal management practice – No Pig Doser
Parameters evaluated:
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% Livability
Weight at weaning (kg/pig)
Weight gain (kg/pig)
ADG (g/day)
% Cull and mortality
Results: Thailand
Globigen®
Control
% Improve
No. of piglet at beginning
379
383
No. of piglet at weaning
349
345
% Livability
92.08
90.08
Weight at beginning (kg/pig)
1.49
1.47
Weight at weaning (kg/pig)
6.02
5.84
+0.18
Weight gain (kg/pig)
4.53
4.37
+0.16
+2
Return on Investment (ROI) ‐ Thailand
Globigen®
Control
Weaning weight (kg/pig)
6.02
5.84
% Livability
92.08
90.08
More weaning piglet per 100 head +2
Weaning Pig price per head ($) 55.40
More Income per 100 head ($)
+110.80
Globigen® Pig Doser cost per 100 head ($)
50
ROI 1 : 2.2
(Not included weaning weight improvement)
Take Home Messages
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Piglets need external support to their immune system:
• Right after birth, before passive protection provided by colostrum
starts working
• The first weeks after weaning, when passive protection decreases
and the animals’ own immune system is still not developed.
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Egg antibodies  Efficient “immune ingredients”
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Decrease of diarrhea, costs for medical treatment, and mortality rates.
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At the end, a healthier piglet will always have the potential to perform to
its full capacity resulting in higher output, lower costs, and therefore
higher profit.
Globigen®
Mind the Immunity gap and use Thank you!
Fellipe Barbosa
EW Nutrition GmbH
Hogenbögen 1
49429 Visbek (Germany)
phone: +49 4445 9868‐0
www.ew‐nutrition.com
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