Effects of Feed Contaminant Deoxynivalenol on Plasma Cytokines and mRNA Expression of Immune Genes in the Intestine of Broiler Chickens
et al. (2013) Effects of Feed Contaminant Deoxynivalenol on Plasma Cytokines and mRNA
Expression of Immune Genes in the Intestine of Broiler Chickens. PLoS ONE 8(8): e71492. doi:10.1371/journal.pone.0071492
Effects of Feed Contaminant Deoxynivalenol on Plasma Cytokines and mRNA Expression of Immune Genes in the Intestine of Broiler Chickens
Josef Bo hm 0
Juliet Spencer, University of San Francisco, United States of America
0 1 Department for Farm Animals and Veterinary Public Health, Institute of Animal Nutrition and Functional Plant Compounds, University of Veterinary Medicine , Vienna , Austria , 2 Department of Animal Hygiene , Behaviour and Management , Faculty of Veterinary Medicine, South Valley University , Qena, Egypt, 3 Clinic for Avian , Reptile and Fish Medicine, Department for Farm Animals and Veterinary Public Health, University of Veterinary Medicine , Vienna , Austria , 4 Department of Biomedical Sciences, Institute of Physiology , Pathophysiology and Biophysics , University of Veterinary Medicine , Vienna , Austria
An experiment was conducted to investigate the individual and combined effects of dietary deoxynivalenol (DON) and a microbial feed additive on plasma cytokine level and on the expression of immune relevant genes in jejunal tissues of broilers. A total of 40 broiler chicks were obtained from a commercial hatchery and divided randomly into four groups (10 birds per group). Birds were reared in battery cages from one day old for 5 weeks. The dietary groups were 1) control birds fed basal diet; 2) DON group fed basal diet contaminated with 10 mg DON/ kg feed; 3) DON + Mycofix group fed basal diet contaminated with 10 mg DON/ kg feed and supplemented with a commercial feed additive, MycofixH Select (MS) (2.5 kg/ ton of feed); 4) Mycofix group fed basal diet supplemented with MS (2.5 kg/ton of feed). At 35 days, the plasma levels of tumor necrosis factor alpha (TNF-a) and interleukin 8 (IL-8) were quantified by ELISA test kits. Furthermore, the mRNA expression of TNF-a, IL-8, IL-1b, interferon gamma (IFNc), transforming growth factor beta receptor I (TGFBR1) and nuclear factor kappa-light-chain-enhancer of activated B cells 1 (NF-kb1) in jejunum were quantified by qRT-PCR. The results showed that the plasma TNF-a decreased in response to DON, while in combination with MS, the effect of DON was reduced. DON down-regulated the relative gene expression of IL-1b, TGFBR1 and IFN-c, and addition of MS to the DON contaminated diet compensates these effects on IL-1b, TGFBR1 but not for IFN-c. Furthermore, supplementation of MS to either DON contaminated or control diet up-regulated the mRNA expression of NF-kb1. In conclusion, DON has the potential to provoke and modulate immunological reactions of broilers and subsequently could increase their susceptibility to disease. The additive seemed to have almost as much of an effect as DON, albeit on different genes.
Funding: This work received the financial support from Biomin GTI GmbH, Herzogenburg, Austria. The funders had no role in study design, data collection and
analysis, decision to publish, or preparation of the manuscript.
Competing Interests: This work received the financial support from Biomin GTI GmbH, Herzogenburg, Austria. This does not alter the authors adherence to all
the PLOS ONE policies on sharing data and materials.
. These authors author contributed equally to this work.
Deoxynivalenol (DON), or vomitoxin, is produced by Fusarium
graminearum (Gibberella zea) and F. culmorum  and considered the
most common contaminant in poultry feedstuffs. It has negative
effects on growth, feed consumption and may induce intestinal
alterations, neurological and reproductive problems . However,
immune impairment is considered the most important outcome of
DON mycotoxicosis . It was shown that DON has both
immunostimulatory and immunosuppressive effects according to
concentration, time and duration of exposure . DON can be
immunotoxic at low dietary concentrations even if there is no
alteration of the productivity traits [3,4,5]. Unfortunately, limited
information is available regarding the immunotoxicity of DON in
poultry. In broiler chickens, DON was shown to suppress the
vaccination response to infectious bronchitis virus (IBV)  and to
Newcastle disease virus (NDV) [6,7]. Recently, DON was shown
to suppress the antibody response to infectious bronchitis vaccine
(IBV) in broiler chickens [8,9]. Furthermore, it was shown that the
dietary inclusion of DON in diets of laying hens resulted in a
reduction of white blood cell number and total lymphocyte
The ability of DON to affect cytokines is important because this
can lead to dysregulation of immune functions. In domestic pigs,
lower IL-1b and IL-8 expression occurred in blood and ileal tissue
after feeding of low doses of DON . Similarly, in broiler
chickens, splenic mRNA expression of IFN-c was down-regulated
as a result of chronic feeding of diets naturally contaminated with
DON and Fusarium mycotoxins .
Cytokines are secreted proteins that regulate the nature of
immune responses by affecting growth, differentiation, and
activation of cells. They are involved in almost all stages of
immunity and inflammation, and cytokine production is induced
Pumpkin seed expeller1
Trace element premix3
1Pumpkin seed expeller is a byproduct of oil manufacture, obtained by pressing
of pumpkin seeds, Cucurbita maxima Duch, moschata (Duch) Poir., Cucurbita
pepo L., and other species of Cucurbita.
2Produced by MIAVIT GmbH & Co. KG, Essen (Oldb.), Germany. Each kilogram of
vitamin premix contains vitamin A, 200,000 IU; vitamin D3, 80,000 IU; vitamin E,
1,600 mg; vitamin K3, 34 mg; vitamin C, 1,300 mg; vitamin B1, 35 mg; vitamin
B2, 135 mg; vitamin B6, 100 mg; vitamin B12, 670 mg; nicotinic acid, 1,340 mg;
calcium pantothenic acid, 235 mg; choline chloride, 8,400 mg; folic acid, 34 mg;
biotin, 3,350 mg; and methionine, 30 g.
3Produced by MIAVIT GmbH & Co. KG, Essen (Oldb.), Germany. Each kilogram of
trace element premix contains calcium, 196 g; phosphorous, 64 g; sodium,
30 g; magnesium, 6 g; copper, 400 mg; zinc, 1,200 mg; iron, 2,000 mg;
manganese, 1,200 mg; cobalt, 20 mg; iodine, 40 mg; selenium, 8 mg.
4Ten milligrams of deoxynivalenol/kg were added to constitute the DON group.
5Two and half kg of MycofixH Select (Biomin GmbH, Herzogenburg, Austria)/ton
of diet were added to constitute the MycofixH group. This plus 10 mg DON/kg
of diet constitutes DON+Mycofix group.
by a variety of stimuli such as viral, bacterial or parasitic infection,
cancer, inflammation, or the interaction between T cells and
antigens. The biological actions of cytokines are produced when
they act as ligands and bind to their high-affinity receptors.
Cytokines can act by an autocrine manner, affecting the cell that
releases the cytokine, or by paracrine manner, affecting nearby
cells, or by an endocrine manner, affecting distant cells after their
distribution via the blood circulation [13,14]. There is a lack of
information about the impacts of DON on the plasma level of
Furthermore, the gastrointestinal tract (GIT) is considered as
important barrier against toxins and contaminants  which has
significant physical, chemical, immunological and microbiological
characteristics. Intestines are large immune organs and have a
broad capability for innate and acquired immune reactions against
various antigens . A variety of soluble mediators have been
shown to participate in the gastrointestinal inflammatory process,
including the pro-inflammatory interleukins . Thus, it
becomes important to investigate the effects of DON on the
relative expression levels of immune genes, including cytokines and
transcription factors, in the intestines of broiler chickens.
Microbial feed additives are frequently used to reduce the
negative impacts of DON. It has been shown that Mycofix can
reduce the negative impact of DON on vaccinal immune response
to IBV, stress index and blood lymphocyte DNA [8,18].
Therefore, the current study was conducted to assess the effects
of five weeks dietary exposure to DON and a microbial feed
additive on the plasma levels of TNF-a and IL-8 and on the
expression of TNF-a, IL-8, IL-1b, TGFBR1, IFNc, TGFBR1 and
NF-kb1 in jejunum of broiler chickens. TNF-a and IL-8 were
selected for investigation in the plasma because both are involved
in the systemic inflammation, activation of the acute phase
reaction, and mediation of inflammatory responses in order to
overcome the infectious antigens. In addition, TNF-a and IL-8 are
known targets of NF-kB, which is modulated in cases of DON
toxicity as a result of activation of mitogen activated protein
kinases . To our knowledge, this is the first in vivo study which
observes these parameters in broiler chickens.
Materials and Methods
The animal experiments were discussed and approved by the
institutional ethics committee of the University of Veterinary
Medicine and Austrian Federal Ministry for Science and Research
under the license number GZ-68.205/0032-II/10b/2010. All
husbandry practices and euthanasia were performed with full
consideration of animal welfare.
Experimental design, birds and diets
A total of forty 1d male broiler chicks (Ross 308) were obtained
from a commercial hatchery and divided randomly into 4 groups
(10 birds per group). Birds of each group received one of the
following dietary treatments; 1) control birds fed basal diet; 2)
DON group fed basal diet contaminated with 10 mg DON/ kg
feed; 3) DON + Mycofix group fed basal diet contaminated with
10 mg DON/ kg feed and supplemented with 2.5 kg of a
commercially microbial feed additive, MycofixH Select (MS),
(Biomin GmbH, Herzogenburg, Austria) per ton of feed; 4)
Mycofix group fed basal diet supplemented with 2.5 kg of a
commercially microbial feed additive, MycofixH Select, (Biomin
GmbH, Herzogenburg, Austria) per ton of feed. The birds were
fed the starter feed from 113 days old and grower feed from 14
35 days old (Table 1). Water and feed were available ad libitum.
Real-time PCR, sense (59-39)
GCAACTATGTTGGACCTGCAAA CCCCTACCCTGTCCCACAA ACCAGGTCCTACTCCAGGAAGAC doi:10.1371/journal.pone.0071492.t002
The climatic conditions and lighting program were
computeroperated and followed the commercial recommendations.
Environmental temperature in the first week of life was 35uC and
reduced to 25uC until the end of the experiment. During the first
week, 22 h of light was provided with a reduction to 20 h
afterward. Representative feed samples were taken at the
beginning of the starter and grower periods and were analyzed
for nutrient and mycotoxin content. Levels of deoxynivalenol,
acetyldeoxynivalenol, zearalenone, nivalenol, and fusarenon-X
were determined in the diets using an HPLC technique .
Measurement of plasma cytokine level
At 35 days old blood samples from all experimental birds were
collected in heparinized tubes, and plasma of each bird was
separated by centrifugation at 1000g for 15 min for determination
of TNF-a and IL-8 levels. The concentrations for tumor necrosis
factor alpha (TNF-a) and interleukin 8 (IL-8) were quantified in
plasma using commercially available ELISA test kits
(Antibodiesonline.com, Aachen, Germany).
Quantification of mRNA of immune genes by
quantitative (q) RT-PCR Sample Extraction
Intestinal tissue samples from mid-jejunum (5 cm from Meckels
diverticulum) from all birds (10 birds/group) were removed
immediately after slaughter (stunning followed by bleeding). The
epithelial layer was collected in Petri dishes on ice by scraping with
a sterile scalpel and immersed in the RNA stabilizing agent,
RNAlater (Qiagen, Hilden, Germany) and frozen at 280uC. After
thawing, 30 mg of the tissue was used to extract total RNA with a
commercial extraction kit (RNeasy Protect Mini Kit, Qiagen,
Hilden, Germany). The RNA amount in extracts was determined
fluorospectrometrically with an EvaGreen RNA determination kit
(Invitrogen, Karlsruhe, Germany). For cDNA synthesis, total RNA
was diluted to 0.2 mg/mL in diethylpyrocarbonate-treated water.
Reverse transcription into cDNA was performed using a High
Capacity cDNA Archive Kit (Applied Biosystems, Darmstadt,
Germany) according to manufacturers instructions included in the
kit. The cDNA was diluted 1 :64 and stored at 220uC until further
Quantitative Real-Time PCR (qRT-PCR)
A commercial master mix (OneStep RT-PCR Kit, Qiagen,
Hilden, Germany) with the addition of EvaGreen as fluorescent
agent (0.08 nmol/L, final concentration) was used for
amplification of GAPDH, TNFa, IL-8, IL-1b, TGFBR1, IFNc, TGFBR1
and NF-kb1. The oligonucleotide primers used for PCR
amplification were synthesized (Table 2).
ACCCACCAAGCTGTGAGCAT TGAGTACTGCGGAGGGTTCAT AAAGCAGACAGGTCCAGCAATAA
The following PCR program was employed on the ViiATM 7
System (Applied Biosystems, Vienna, Austria) to amplify target
mRNA in tissue extracts: initial denaturation for 15 min at 95uC
(1 cycle); followed by 45 cycles of 30 sec 95uC and 50 sec 63uC. A
dissociation curve was generated after 45 cycles in order to
determine melting points of the amplified cDNA.
Calculation of relative mRNA Expression
GAPDH was investigated as a reference gene based on
expression stability. The expression of genes of interest (GOIs)
was normalized using the housekeeping gene GAPDH. Data were
analyzed using the efficiency corrected Delta-Delta-Ct method
. The mRNA expression of TNFa, IL-8, IL-1b, TGFBR1,
IFNc, and NF-kb1 were evaluated in jejunum.
The statistical program SPSS (version 17; SPSS GmbH, SPSS
Inc., Munich, Germany) was used for data analysis. The
Kolmogorov-Smirnov test was used to test the normal distribution
of the data. An ANOVA was performed between the 4 groups,
followed by Duncan test to find the significance between dietary
treatments. The probability values of 0.05 (P#0.05) were
Mycotoxin contents of diets
The dietary concentration of DON in the starter control feed
was 506680 mg/kg, and the level of the zearalenone was under
the limits of detection. In the DON-contaminated starter feed the
concentration was 10,509680 mg/kg, and other mycotoxins were
under the limits of detection. In the DON non-contaminated
control grower diet, DON concentration was 611695 mg/kg and
in the DON contaminated grower diet; DON was 10,614695 mg/
kg. Other mycotoxins were under the limits of detection.
Plasma cytokines production
In plasma, TNF-a was significantly down-regulated in broiler
chickens receiving DON (P,0.05) compared to the control group
(Table 3). Moreover, addition of MS counteracted the effect of
DON on plasma TNF-a and the value was comparable to that of
control birds. Differently, IL-8 did not significantly alter (P.0.05)
for all the broiler chickens receiving DON, counteracting agent or
DON and the counteracting agent compared to the control group.
Relative mRNA expression of immune genes
DON down-regulated (P,0.001) the mRNA relative expression
of IL-1b in jejunal tissue. However, feed supplementation with the
counteracting agent was efficient to increase the mRNA relative
expression of IL-1b to values that were comparable to control
birds (Figure 1B). Furthermore, DON feeding resulted in a trend
for down-regulation (P,0.1) of TGFBR-1 in the jejunal tissue of
broilers (Figure 1B). However, Mycofix addition to either DON
contaminated diet or basal control diet did not produce a
significant change on the relative gene expression of TGFBR-1
and IL-1 (Figure 1B). Moreover, the relative mRNA expression
of IL-8 was up-regulated (P,0.05) when Mycofix was added to
DON contaminated diet (P,0.05; Figure 1A). Interferon-gamma
(IFN-c) relative gene expression was significantly down-regulated
(P,0.001) due to DON feeding, as well as when Mycofix was
added to either DON contaminated feed or to basal control feed
Mycofix supplementation either with or without DON
significantly (P,0.05) up-regulates the expression of NF-kB1
(Figure 1C). Furthermore, the relative mRNA expression of
TNF-a also up-regulated (P,0.05) due to Mycofix
supplementation and tended to be up-regulated (P,0.1) when MS was added
to DON contaminated feed (Figure 1A).
The impairment of immune function is the most important toxic
effect of DON and may result in either immune-stimulation or
immune-suppression depending on the dose, time and duration of
exposure. The results of this study represent new information
regarding the impacts of deoxynivalenol oral exposure on plasma
cytokines and expression of immune genes, including the genes
that regulate the production of pro-inflammatory cytokines in the
gastrointestinal tract of broiler chickens.
DON is known to either suppress or stimulate immunological
parameters such as inhibition of lymphocyte proliferation with a
concomitant elevation of immunoglobulin and cytokine
concentrations in vitro or in vivo, even at permissible levels [3,4,21,22]. The
phenomenon that protein synthesis inhibitors up-regulate cytokine
gene expression and secretion is called superinduction and may
be due to inhibition of particularly labile translational repressor
proteins [23,24,25]. However, it should not be ignored that
immunotoxicity studies have focused mainly on the mouse model
with only few investigations in domestic animals. Therefore, the
current experiment was conducted to investigate the impacts of
DON and a microbial feed additive on plasma levels of cytokine
TNF-a and interleukin 8 (IL-8) and on the relative gene expression
of TNF-a, IL-8, NF-kB1, IFN-c, IL-1b, and TGFBR1 in the
jejunum of broilers.
Interleukins are a group of cytokines that are important
components of the immune system. They play a physiological
role in inflammation and a pathological role in systemic
inflammatory states, and any imbalance in cytokine production
and dysregulation of a cytokine process could result in various
pathological disorders . In the present study, plasma level of
IL-8 showed less change due to the addition of either DON or
Mycofix Select but these changes were not statistically significant.
Oral DON decreased the production of IL-8 which mediates the
inflammatory immune response, suggesting that DON ingestion
could impair immune function and elevate susceptibility to disease.
On the other hand, addition of Mycofix Select to either DON
contaminated diet or to basal control diet resulted in a smaller
increase of IL-8 production, indicating that this feed additive
improved the immune response of chickens, probably due to its
immune stimulating properties.
The current state of knowledge indicates that DON can have
marked immunological effects in chickens. Feeding of 10 mg
DON/kg feed to broilers decreased the plasma concentration of
TNF-a in the present study. TNF-a is a cytokine involved in
systemic inflammation and stimulation of the acute phase reaction.
It is produced chiefly by activated macrophages (M1), although it
can be produced by a variety of cells such as CD4+ lymphocytes,
NK cells, and neurons. Large amounts of TNF-a are released in
response to lipopolysaccharide, other bacterial products, and other
cytokines such as interleukin (IL)-1, IL-2, IFN-c, IFN-a,
Granulocyte Macrophage Colony Stimulating factor (GM-CSF), the
Transforming Growth Factor (TGF)- .
The primary role of TNF is in the regulation of immune cells. It
is possible that DON exposure resulting in decreased levels of
TNF-a contributes to impairment of immune function in poultry
and increased susceptibility to infectious diseases when birds are
fed DON for long periods. However, this hypothesis needs further
studies to be confirmed. In contrast, feeding of DON
contaminated diet (2.9 mg/kg feed) did not affect the TNF a level in the
plasma of pigs , and pigs that received both LPS and DON
induced TNF-a compared with controls. The difference between
chickens and pigs in the term of sensitivity to DON toxicity, the
dose, and period of exposure could explain the difference between
both studies regarding TNF-a level in the plasma. However, in
comparison to pigs that received LPS alone, DON and LPS pigs
had lower levels of plasma TNF a, indicating that DON reduced
plasma TNF a in the presence of LPS . In the current study,
the addition of a microbial feed additive counteracted the reducing
level of TNF-a in the plasma of broiler fed DON, suggesting that
this feed supplement was beneficial for the reduction of DON
effects on plasma cytokines. Furthermore, birds receiving DON
and/ or Mycofix Select appeared clinically healthy without any
clinical signs during the whole experimental period and their daily
body weight gain did not differ between dietary treatments.
It was shown that DON is able to phosphorylate Ikb in human
Caco-2 cells , resulting in release of the transcription factor
(Nuclear Factor kappa b, NF-kb) from its inhibitor Ikb, which
then allows NF-kb to translocate into nucleus and activate the
transcription of specific genes . Deoxynivalenol was found to
increase the binding activity of NF-kb in vitro  through
inhibition of resynthesis of IkBa, a member of Ik family.
Nuclear Factor kappa b (NF-kb) is a transcription factor that is
activated by various intra- and extra-cellular stimuli such as
cytokines, oxidant-free radicals, and bacterial or viral products.
NF-k stimulates the expression of genes involved in a wide
variety of biological functions. It is located normally and in a
steady state in the cytoplasm in association with the inhibitory Ik
protein in an inactive form . The activated transcription factor
(NF-kb ) then translocates to the nucleus of the cell
[33,34,35,36,37] and switches on gene expression for cytokines
(i.e. activates the transcription of pro-inflammatory genes) and
subsequent release of proinflammatory mediators  such as
tumor necrosis factor-a (TNF-a) and interleukins .
In this study, addition of DON caused a significant reduction of
mRNA levels of some genes (IL-1b, TGFBR1, IFN-c) but caused
no changes in others (TNF-a, IL-8, NF-kb). This is in contrast to
DON action in the usual mouse model but fits with DON action in
pigs. In mice, it was found that the levels of mRNA of IL-1, IL-6,
and TNF-a, IFN-c, IL-4 and IL-10 were increased after a single
oral dose of 5 and 25 mg/kg body weight . The capability of
DON to affect the cytokine gene expression is important because it
can indicate the modulation of immune functions. It was shown
that up-regulation of the relative gene expression of NF-kb 1,
TNF-a and IL-8 in case of DON toxicity due to the decrease of
the protein synthesis in the intestinal cells [23,24,25]. However, in
domestic pigs, lower IL-1b, IL-8 and TNF-a expressions occurred
in blood and ileal tissue after feeding of low doses of DON ,
suggesting the down-regulation of immune-related transcription
factors and pro-inflammatory immune factors of pigs fed DON.
Similarly, feeding of low (3.44 mg/ kg feed), medium (6.30 to
10.68 mg/kg feed), and high (16.97 mg/kg feed) deoxynivalenol
for 3 weeks experiment did not alter IL-8 expression in the cecal
tonsils of broilers .
In contrast, IL-1b and IFN-c was down-regulated after oral
DON exposure in the present experiment. This result is in
agreement with the previous report of Cheng et al. ,which
found that in pigs treated with DON for 6 weeks, a decrease in the
expression of IFN-c, IL-1b was observed. Similarly, in broiler
chickens, splenic mRNA expression of IFN-c was down-regulated
as a result of chronic feeding of naturally contaminated diets with
DON and other Fusarium mycotoxin contaminated diet . In
another studies, IFN-c gene expression was up-regulated in the
cecal tonsils of chickens fed Furarium mycotoxin challenged with
coccidia . In this context, it becomes evident that DON has a
modulating effect on the innate immune response. Moreover, the
decline of IFN-c expression may impair the anti-virus ability of the
host. Cell-mediated and humoral immune response network are
also interfered by a decrease of IL-1b level. Cytokines play an
important role in the network of immune responses and account
for communication, activation, maturation and differentiation
among immune cells. Since dietary DON can modify gene
expression of cytokines, it may impair disease resistance in poultry.
Furthermore, DON had a similar down-regulating effect on
transforming growth factor beta receptor I (TGFBR1), which is a
cytokine protein secreted by many cell types, including
macrophages. TGFBR1 plays a crucial role in the regulation of the cell
cycle and regulates a variety of different cellular developmental
processes including growth, differentiation, proliferation, and cell
death. TGF-b is believed to be important in regulation of the
immune system by regulatory T cell and in the differentiation of
both regulatory T cell and Th17 cells. TGF-b appears to block the
activation of lymphocytes and monocyte derived phagocytes. In
the current study, DON down-regulated this cytokine in the
jejunal tissue of broilers. In addition, a microbial feed additive
when added to DON contaminated diet counteracted the immune
modulation of DON on TGFBR1, and IL-1b. However, it has no
counteracting effects on down-regulation of the relative mRNA
expression of IFN-c caused by DON. The ability of Mycofix Select
to reverse the effects of DON for some genes but not others could
be related to the dose of the feed additive added to DON
contaminated diet. In this study, 2.5 kg Mycofix Select per ton of
diet was used, which could not be enough to reverse all the effects
of of 10 mg DON/ kg feed. Interestingly, Mycofix in and of itself
increased the levels of several mRNAs that were not affected by
DON (.TNF-a, IL-8 and NF-kB), This indicates that addition of
feed additive Mycofix Select stimulates the immune response
genes, suggesting that it has an immune stimulating property. This
information may be important as Mycofix Select is used
commercially as antidote for mycotoxin.
In conclusion, DON reduced the level of plasma TNF-a and
down-regulated the relative mRNA expression of IFN-c, IL-1b,
and TGFBR1, which indicates the ability of DON to inhibit the
protein synthesis. However, mRNA relative expression of IL-8,
NF-kB and TNF-a were up-regulated, suggesting that DON could
have an effect on the innate immune response which can impair
the resistance of chickens to infectious diseases and consequently
increase the susceptibility of the host to infection. Furthermore, a
microbial feed additive has the ability to modulate the DON
effects on plasma level of TNF-a and on the relative expression of
mRNA of IL-1b and TGFBR1 in the intestinal epithelium of
This work received the financial support from Biomin GTI GmbH,
Herzogenburg, Austria. The funders had no role in study design, data
collection and analysis, decision to publish, or preparation of the
manuscript. We would like to thank Georg Hofer (Institute for Physiology,
Pathophysiology and Biophysics, University of Veterinary medicine,
Vienna, Austria) for his skillful technical assistance.
Conceived and designed the experiments: KG WA JB. Performed the
experiments: KG WA SS CS. Analyzed the data: KG WA AS SS.
Contributed reagents/materials/analysis tools: KG WA CS SS AS JB.
Wrote the paper: KG WA JB.
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