Effects of beta-1,3-glucan (AletaTM ) on vaccination response in broiler
chickens
G. Horst,∗,1 R. Levine
∗
Kemin Industries, Inc., Des Moines, IA 50317, USA; and † Southern Poultry Research Group, Inc.,
Watkinsville, GA 30677, USA
ABSTRACT This 42-day study evaluated the effects
of dietary supplementation with β -1,3-glucan (AletaTM )
on the vaccination response to Newcastle disease virus
(NDV), avian infectious bronchitis virus (IBV), and in-
fectious bursal disease (IBD) in a non-challenged envi-
ronment. This trial included 600 chicks (all vaccinated
with IBD at the hatchery) which were assigned to 1 of
3 treatments: vaccination (NDV, IBV), no vaccination,
or vaccination combined with feed supplemented with
Aleta (100 g/MT of feed). The vaccination with Aleta
treatment group showed a trend for improved FCR
that was not statistically significant. Control birds that
were not vaccinated for IBV had significantly lower IBV
Key words: beta glucan, broiler, Newcastle, vaccine
INTRODUCTION
Until recently, the prophylactic supplementation of
animal feed with antibiotics was common. This prac-
tice led to reduced incidence of some diseases, increased
growth rates, and improved feed efficiency in poultry
and other livestock. There is now, however, increasing
evidence that such long-term, and nonspecific, exposure
to antibiotics leads to bacterial resistance with signif-
icant impacts on the future efficacy of these critical
drugs (Aarestrup, 2005). Consequently, several coun-
tries have restricted or banned the prophylactic use of
antibiotics in animal feed. This gives a new urgency to
the search for alternative strategies to maintain animal
performance and protect animals from disease.
One promising strategy is the use of immune mod-
ulators such as beta-1,3-glucan to bolster an animal’s
innate immunity (Chan et al., 2009; Barsanti et al.,
2011). When animals are exposed to beta glucan, their
immune system detects the molecule as a pathogen-
associated molecular pattern, which triggers a cascade
of immune system responses. This response has evolved
over time since beta-1,3-glucan is found in the cells of

C 2018 Poultry Science Association Inc.
Received September 18, 2018.
Accepted November 8, 2018.
1
Corresponding author: geoff.horst@kemin.com
,∗ R. Chick, and C. Hofacre†
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titers on day 21 compared to birds that were vaccinated
(both with and without Aleta). Surprisingly, there was
significant separation among treatment groups for NDV
titer levels, especially on day 21, where birds vaccinated
and supplemented with Aleta had significantly higher
titer levels compared to vaccination alone or no vacci-
nation at all. Critically, only 14% of the birds receiving
the vaccine plus Aleta had titer levels below the critical
titer threshold for immunity compared to 28% of the
birds receiving the vaccine alone and 40% of the unvac-
cinated birds. This suggests that Aleta supplementation
may help to improve the vaccination response by birds,
especially for NDV.
2018 Poultry Science 0:1–5
http://dx.doi.org/10.3382/ps/pey523
potentially pathogenic organisms such as fungi, yeast,
and bacteria.
In poultry, supplementing feeds with beta-1,3-glucan
has shown to increase immune cell activation (Guo
et al., 2003; Chae et al., 2006; Lowry et al., 2005)
and immune cell recruitment to the gut (Levine et al.,
2018). Subsequently, several groups have demonstrated
improved resistance to bacterial infections (Lowry et
al., 2005; Huff et al., 2006; Chen et al., 2008), improved
intestinal health following coccidiosis infection (Cox
et al., 2010; Levine et al., 2018), and overall improve-
ments in growth rates and feed conversion efficiency
(Chae et al., 2006; An et al., 2008; Zhang et al.. 2008;
Morales-Lopez et al., 2009; Cox et al., 2010; Moon et
al., 2016; Levine et al., 2018).
Vaccination is another key tool to help protect an-
imals from certain types of diseases, especially viral
diseases such as infectious bronchitis (IBV), infectious
bursal disease (IBD), and Newcastle disease (ND).
However, there are not vaccines for all major diseases
and the response to certain vaccines, especially Newcas-
tle disease virus (NDV), can be variable, which leaves
some birds unprotected and can put a flock at risk of
an outbreak (van Boven et al., 2008; Botus et al., 2010;
Hossain et al., 2010; Ghahramani et al., 2014). Previ-
ous studies to determine potential synergistic effects of
yeast-based beta glucans and vaccines have been mixed:
Cheng et al. (2004) showed no effect on ND titers,
1
2 HORST ET AL.
whereas An et al. (2008) showed significant increases
in both ND and IBV titers in birds supplemented with
beta glucan.
In this study, we examine whether supplementation
with AletaTM (a highly-concentrated beta-1,3-glucan
product) can improve the immune response to NDV
and IBV vaccination and improve performance metrics
in a 42-day pen trial.
MATERIALS AND METHODS
Experimental Design
In this 12-pen study, 600 chicks were assigned to 3
treatment groups with 4 replicate pens, with 50 birds
allocated per pen. Treatment groups were assigned to
pens using randomized complete block design. Southern
Poultry Research Group completed randomization and
assignment of treatment groups to pens using random
permutation tables (Cochran and Cox, 1992).
Treatment Groups
1. Control feed (no AletaTM ), no IBV or ND vaccine
2. Control feed (no AletaTM ), vaccinated for IBV and
ND
3. Control feed with 100 ppm AletaTM , vaccinated for
IBV and ND
Birds
A total of 600 day-of-hatch Aviagen straight run
broiler chicks were obtained from Baldwin, GA. Birds
were non-sexed and received routine vaccinations
(HVT-IBD [Vaxxitek, R Boehringer Ingelheim]), and
breeder flock number information was recorded at the
hatchery. All animal handling procedures complied with
the Institutional Animal Care and Use Committee. The
study began when day-of-hatch birds were placed in
treatment groups on day 0 and allocated to experimen-
tal pens. Only healthy appearing birds were allocated
for study use, and the final number and disposition of
all birds not allocated was documented. No birds were
replaced during the study. Each bird was also tagged
and used as the unit of measure for virus titers. Pen
security prevented bird migration.
All birds were monitored for general flock condi-
tion, temperature, lighting, water, feed, litter condition,
unanticipated house conditions/events, and mortality;
findings were documented twice daily during the reg-
ular working hours. Birds were culled only to relieve
suffering. The date and removal weight were recorded
for any bird culled (or found dead), gross necropsy was
performed on all culled (or dead) birds, and gender and
probable cause of death were recorded. All birds were
disposed of by appropriate methods. All mortalities and
remaining feeds (including mixer flushes) were buried in
the Southern Poultry Research Group on-site disposal
pit.
Housing and Environmental Control
At study initiation, 50 broiler chicks were allocated
to 12 floor pens measuring 5 × 10 (1.0 ft2 /bird stock-
ing density) in a modified conventional poultry house
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(fan-cooled) with solid sides and dirt floors. Thermo-
statically controlled gas heaters were the primary heat
source with supplemental heat lamps (one lamp per
pen) providing heat when needed. Birds were raised
under ambient humidity and were provided 24-h light-
ing. At placement each pen contained approximately 4
inches of fresh pine shavings. Litter was not replaced
during the study course. Each pen contained 1 tube
feeder and 1 bell drinker resulting in a 50-bird/feeder
and drinker ratio. Feed and water were consumed ad
libitum throughout the trial.
Diet
Rations were fed as follows: starter diet day 0 to 21,
grower diet day 21 to 35, and finisher diet day 35to
42. Diets were fed as crumbles (starter diet) or pel-
lets (grower and finisher diets). On day 21, 35, and
42, all previous feed was removed from each pen, in-
dividually weighed, discarded, and replaced (day 21
and 35) or discarded (day 42). Feed formulations for
this study consisted of unmedicated commercial-type
broiler starter and grower diets compounded with com-
monly used United States feedstuffs representative of
local formulations, and calculated analyses met or ex-
ceeded NRC standards.
Experimental treatment feeds were prepared from a
basal starter and grower feed with quantities of all basal
feed and test articles used to prepare treatment batches
documented. Treatment group 3 contained the basal
diet supplemented with 100 g/MT of AletaTM (Kemin
Industries, Inc., Des Moines, IA). Aleta is dried Eu-
glena gracilis, a single-celled organism, which contains
approximately 50% beta glucan by weight. To assure
uniform distribution of the beta glucan, pre-pelleted
treatment feeds were mixed at the Southern Poultry
Research feed mill and pelleted in a California Pellet
mill at 80◦ C (with pellet temperature recorded). After
mixing was completed, feed was distributed among pens
of designated treatment groups.
Vaccination
On arrival at Southern Poultry Research Group and
before placement, all chicks in treatment groups 2 and
3 were eye dropped with one dose of ND vaccine (C2)
and one dose of IBV vaccine (GA98). A boost was re-
peated by eye drop at 21 d of age to all birds. No con-
comitant drug therapy was used during the study. To
prevent cross-contamination, plastic disposable boots
 BETA GLUCAN EFFECTS ON BROILER VACCINATION 3
were worn when entering pens and changed between to the low number of replicate pens (4) per treatment
each pen. group and the fact that the birds were not challenged
with either a biological or environmental stressor to dra-
matically affect growth.
Performance Parameters Birds treated with vaccine alone had numerically
Bird weights by pen were recorded on day 0 and day lower mortality (2%) compared to unvaccinated birds
42. Thirty birds per treatment group were bled on day (6%) and vaccinated birds fed Aleta (5%) (Table 1).
7, 21, and 42 for the serology study. Serum was stored Birds treated with vaccines (with and without supple-

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in a freezer until trial termination. All serum was tested
for ND, IBV, and IBD by ELISA at one time with the
same IDEXX kit (IDEXX, Westbrook, ME).
Statistical Analysis
Mortality, weight gain, and feed conversion metrics
were analyzed by ANOVA using individual pens as
replicates. Viral titers were not normally distributed
after Log2 distributions, especially for NDV and IBV
titers. Therefore, titer statistical comparisons among
treatment groups (using individual birds as replicates)
were performed using the non-parametric Kruskal-
Wallis rank test followed by Conover post hoc pairwise
comparisons which corrects for multiple comparisons
(Sokal and Rohlf, 2001).
RESULTS AND DISCUSSION
Performance Parameters
Overall, there were no significant differences in mor-
tality, weight gain, or feed conversion between treat-
ment groups (Table 1). This could be expected due
mented Aleta) showed modest but not significant in-
creases in weight gain compared to the control group.
Vaccinated birds supplemented with Aleta had numer-
ically the best feed conversion efficiency, showing a 2.4-
point improvement (i.e., 0.024 reduction in feed con-
version ratio) over the control group and a 3.6-point
improvement over the vaccine alone group (Table 1).
Serological Data
Virus titers for NDV, IBD, and IBV displayed the
typical time-dependent pattern of high titer levels at
day 7, reflecting the high amount of maternal antibodies
still in circulation. Titer levels declined precipitously by
day 21 and then climbed again by day 42, reflecting the
bird’s natural antibody production (Table 2).
Aleta had no impact on IBD titer levels across all
time points (Table 2). According to facility personnel,
the IBD maternal antibody (mAb) titers at Fieldale
tend to be very high and this is reflected in the high
day 7 titers across all treatment groups (>4,000 or
Log2 >12). The mAb declined by day 21, and the
recombinant IBD vaccine titers began to rise by day
42. Since the IBD insert is in the Herpes virus (HVT)

Table 1. Summary of bird performance (day 0 to 42).

Treatment

(–) vaccine (+) vaccine (+) vaccine

Performance criteria (–) Aleta (−) Aleta (+) Aleta
Feed conversion (kg feed/kg weight gain) 1.645 (0.005) 1.657 (0.021) 1.621 (0.010)
Average weight gain (kg) 2.222 (0.031) 2.245 (0.041) 2.239 (0.061)
Mortality (%) 6.5 (0.2) 2.0 (0.1) 5.0 (0.1)

Means (+/− SE) of 4 replicate pens per treatment group.

Table 2. Mean (Log2 ), median (Log2 ), and coefficient of variation (CV) of titer to Newcastle disease virus (NDV), avian infectious
bronchitis virus (IBV), and infectious bursal disease (IBD) by treatment group on day 7, 21, and 42 (n = 120 birds per treatment
group).

Day 7 Day 21 Day 42

Treatment Mean Median CV Mean Median CV Mean Median CV

NDV No vaccine 8.86 9.08 0.24 3.44 4.25c 0.89 7.99 8.29b 0.24
Vaccine only 8.65 8.97 0.24 4.40 5.00b 0.72 8.76 8.82a 0.14
Vaccine w/Aleta 8.17 8.33 0.30 5.54 6.25a 0.47 8.85 8.91a 0.13
IBD No vaccine 12.40 12.62 0.05 8.40 8.36 0.14 12.28 12.47 0.08
Vaccine only 12.35 12.47 0.05 8.40 8.57 0.16 12.62 12.67 0.05
Vaccine w/Aleta 12.35 12.43 0.05 8.30 8.35 0.16 12.40 12.57 0.07
IBV No vaccine 10.85 10.91 0.09 4.53 5.25b 0.60 6.94 7.39 0.33
Vaccine only 10.60 10.54 0.10 5.93 6.17a 0.46 7.26 7.50 0.29
Vaccine w/Aleta 10.58 10.55 0.10 5.88 6.51a 0.51 6.67 7.19 0.37
a–c
Medians with different superscripts represent significantly different populations between treatment groups using the Kruskal–Wallis rank test
followed by Conover post-hoc comparisons.
4 HORST ET AL.

which primarily replicates in T cells (after an initial B

cell), the immunity normally develops slowly to IBD.
The mAb for IBV were still present at day 7 and
began to drop by day 21 (Table 2). Since the primary
immunity to IBV is local (Harderian Gland/BALT), it
is not expected that measuring circulating antibodies in
serum by ELISA will result in elevated titers as com-
monly seen with IBD or NDV. IBV is highly contagious

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and despite facility personnel changing boots between
pens, it appears the IBV moved into the non-vaccinated
pens. Vaccinated birds, both with and without Aleta,
had significantly higher titers and narrower coefficient
of variation (CV) than the unvaccinated control group,
as expected (Table 2). However, there did not appear
to be a strong impact of Aleta in further enhancing the
titer levels for IBV in combination with vaccination.
NDV titers were most responsive to vaccination, both
with and without supplementation of Aleta. On day 21,
there was significant separation among the 3 treatment
groups, with the vaccinated group supplemented with
Aleta demonstrating the highest titer levels followed by
vaccine alone and the control group with the lowest
titer levels (Table 2 and Figure 1). The coefficient of
variation was also different between groups, with the
control group having the highest variation (0.89), fol-
lowed by the vaccine only group (0.72), and finally the
vaccinated birds supplemented with Aleta (0.47). Ide-
ally, CVs should be less than 0.50 to be considered an
effective and homogenous vaccination.
With closer inspection of the distribution of vaccine
titers among treatment groups, it is apparent that the
distribution is far from normal, even with Log2 trans-
formation (Figure 2). In fact, some proportion of the
birds had non-detectable titer levels, far below the as-
sumed Log2 NDV titer of >3 in order to have pro-
tection against ND (van Boven et al., 2008; Ghahra-
mani et al., 2014). During the critical time period when
maternal antibodies are declining and the intrinsically
produced antibodies are still ramping up, many of the
birds have titer levels dip below this critical threshold.
In the unvaccinated group, 40% of the birds fell below
this threshold and with vaccination alone, about 28%
Figure 1. Box plot of NDV titers (Log2 ) from 120 birds per treat-
ment group on day 7, 21, and 42. Treatment groups with different su-
perscripts represent significantly different populations between treat-
ment groups using the Kruskal–Wallis rank test followed by Conover
post hoc comparisons.
Figure 2. Relative frequency histogram of NDV titers (Log2) from
120 birds per treatment group on day 21. Treatment groups with
different superscripts represent significantly different populations be-
tween treatment groups using the Kruskal–Wallis rank test followed
by Conover post hoc comparison.
Figure 3. The percentage of birds from each treatment group that
exceed the NDV titer threshold of Log2 > 3 presumed required to confer
immunity.
of birds were still susceptible. By stark contrast, the
vaccination plus Aleta group had only 14% of the birds
fall below this critical titer threshold (Figure 3), which
is biologically significant since previous studies indicate
that herd immunity for NDV occurs when 85% or more
of the population is above the critical titer threshold
(van Boven et al., 2008).
The effect of vaccination on NDV titer levels contin-
ued to be apparent on day 42, with both vaccinated
groups having significantly higher titer levels and nar-
rower CVs compared to the unvaccinated control, but
there was no longer any separation between the vaccine
alone and vaccine with Aleta groups (Table 2, Figure 2).
By day 42, nearly all birds had titers above the criti-
cal threshold and therefore differences among groups
were unlikely to have an impact on disease prevention
by that time period.
In this study, NDV titers were the most variable
among all the virus antibodies tested. Specifically, the
unvaccinated control group was characterized by a high
coefficient of variation and, importantly, a large pro-
portion of birds with low to no observable NDV titers
on day 21, meaning they could be susceptible to ND
leading to a potential outbreak in the population. Vac-
cination alone did significantly increase titers levels and
decreased CV, but still a surprisingly high 28% of the
population was below the critical titer levels to con-
fer immunity. It was only with the vaccine plus Aleta
 BETA GLUCAN EFFECTS ON BROILER VACCINATION
supplementation that 86% of the population reached
the critical titer threshold, which is above the theoret-
ical 85% threshold for herd immunity from ND. Prior
research has shown that beta glucans can increase in-
nate immune activity while also prime critical immune
cells involved in adaptive immunity including both B-
and T cells (Guo et al., 2003); this may be a potential
mechanism for improving vaccine responsiveness seen
among birds supplemented with Aleta in combination
with normal vaccination protocols.
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