ORIGINAL ARTICLE
Inclusion of Bacillus amyloliquefaciens strain TOA5001
in the diet of broilers suppresses the symptoms of
coccidiosis by modulating intestinal microbiota
Takamitsu TSUKAHARA,
1
Ryo INOUE,
2
Keizo NAKAYAMA
1
and Takio INATOMI
3,4
1
Kyoto Institute of Nutrition & Pathology, Ujitawara,
2
Kyoto Prefectural University, Shimogamo, Kyoto,
3
TOA
Pharmaceutical Co. Ltd., and
4
Inatomi Animal Hospital, Tokyo, Japan
ABSTRACT
Coccidiosis is an intestinal parasitic infection and one of the most prevalent and economically damaging diseases of
chickens. Furthermore, coccidia-induced mucogenesis promotes secondary colonization by Clostridium perfringens,a
major pathogen of chickens that causes necrotic enteritis. Our previous work found that supernatant of a culture of
Bacillus amyloliquefaciens strain TOA5001 (BA) inhibited the growth of C. perfringens on Gifu anaerobic broth med-
ium. Accordingly, we evaluated the effectiveness of dietary BA administration in inhibiting C. perfringens colonization
of the intestine in broilers that were experimentally infected with coccidia. Ten healthy broilers from a BA-supplemen-
ted (2 9 10
5
colony-forming units/g of feed) broiler group and 10 from a non-treated group were challenged with
Eimeria tenella and E. maxima (5000 oocysts of each species/chick) at 28 days old. At 36 days old, five chicks from
each group were slaughtered, whereas the remaining five in each group were killed at 49 days old. Dietary BA admin-
istration into Eimeria-challenged birds reduced coccidial symptoms such as intestinal lesions. It also modified the cecal
microbiota through suppressing C. perfringens and E. coli colonization, and inducing domination of Faecalibacterium
prausnitzii, the Lactobacillus group and unknown Lachnospiraceae genera by bacterial DNA-based metagenome analy-
ses. B. amyloliquefaciens TOA5001 supplementation suppressed the symptoms of coccidiosis by modulating cecal
microbiota in Eimeria-challenged broilers.
Key words: Bacillus amyloliquefaciens, broiler, cecal microbiota, Clostridium perfringens, coccidial infection.
INTRODUCTION
Coccidiosis is an intestinal parasitic infection and one of
the most prevalent and economically damaging dis-
eases of chickens. Parasitic protozoa of the genus Eime-
ria infect the intestinal tract and cause tissue damage,
resulting in interruption of feeding and digestive pro-
cesses, poor nutrient absorption, dehydration, blood
loss and increased susceptibility to other disease agents
(Morris & Gasser 2006). The clinical signs of coccidiosis
include mortality, morbidity, diarrhea and bloody feces
(Williams 1999). However, control and treatment of
the disease increases production costs (Hafez 2008).
Indeed, anti-coccidial drugs and live vaccines are the
main control measures for coccidiosis, but there are
increasing problems with prolonged drug usage and
the high cost of vaccines. Therefore, alternative strate-
gies are needed for safety and cost effectiveness of coc-
cidiosis treatment in chickens (Dalloul et al. 2006;
Williams 2006).
Probiotics have been among the most promising
alternative coccidiosis control strategies investigated
(Dalloul et al. 2003; Lee et al. 2007). For example, in
a previous study conducted in our laboratory, a diet-
ary probiotic mixture containing Bacillus mesentericus,
Clostridium butyricum and Streptococcus faecalis adminis-
tration was effective in preventing coccidial infection
in broilers (Hayakawa et al. 2014). The effectiveness
of this probiotic against coccidiosis was associated
with stimulation of total immunoglobulin A (IgA)
production in the intestine, and elongation of the
villous height in the jejunum. Bacillus amyloliquefa-
ciens strain TOA5001 (BA), a spore-forming bac-
terium, produces the anti-fungal factor Bacillomycin
Lc, and the culture supernatants of this strain were
Correspondence: Takamitsu Tsukahara, Kyoto Institute of
Nutrition & Pathology, Ujitawara, Kyoto 610-0231, Japan.
(Email: tsukahara@kyoto-inp.cc)
Received 9 May 2017; accepted for publication 17
November 2017.
© 2017 Japanese Society of Animal Science
Animal Science Journal (2017) , – doi:10.1111/asj.12980