Poultry USA - January 2020 - 25

Assessment of safety and heat stability
under commercial conditions of the probiotic
strain Bacillus amyloliquefaciens CECT 5940
(Ecobiol®)
Anita Menconi1, Stella Molck2, Bayo Sokale1, and Mark Daniel1
1: Evonik Corporation, USA; 2: Evonik Nutrition & Care GmbH

Direct-Fed Microbials (DFMs) have been
an important functional feed additive in the
Veterinarian's and Nutritionist's "toolbox" for the
maintenance of intestinal health in poultry. One
critical aspect pertaining to the use of DFMs is the
close attention that must be given to the entire
process of selecting a strain, which encompasses
the knowledge on the strain safety and genetic
assessment, antimicrobial susceptibility, application
potential, survivability, functionality under research
and commercial conditions, and different modes
of action. Under commercial conditions, each one
of the above-mentioned aspects must fall in place
to assure the delivery and functionality of the live
beneficial microorganisms chosen for that specific
poultry flock.
Although several species of Bacillus used as
DFMs are generally recognized as safe, individual
strains could contain genes conferring resistance
to antibiotics and/or have the potential for toxin
production and hemolytic effects (Marteau, 2001;
Pedersen et al., 2002; Doron and Snydman,
2015). Consequently, it is crucial to assess and
validate the safety and characteristics of strains
considered for use in livestock production. In
addition to safety related factors, the stability
and survival rate of strains can vary among the
same species, which may influence commercial
feeding applications. Therefore, the purpose of
this paper is to describe safety and antimicrobial
susceptibility related studies, and, as the next
step into commercial testing, to demonstrate
the stability of the strain of Ecobiol (Bacillus
amyloliquefaciens CECT 5940) under commercial
feed manufacturing processes.

ASSESSMENT OF STRAIN SAFETY
A bioinformatic study based on the whole genome
sequence (WGS) was conducted to investigate
the absence of genes encoding enterotoxins
(nhe, hbl, and cytK) and cereulide synthase (ces)
in B. amyloliquefaciens CECT 5940. Briefly, the
genome sequencing of the above-mentioned strain
was performed using the whole genome shotgun
method. Data generated by Illumina and SMRT
sequencing was used to acquire the complete
circular genome sequence of B. amyloliquefaciens
CECT 5940. A BLAST analysis was used to
exclude the presence of the following genes
encoding relevant toxins: cytK (single protein
cytotoxin K), hblA, hblB, hblC, and hblD (hemolysin
BL, four protein components), nheA, nheB, and
nheC (non-hemolytic enterotoxin, three protein
components) and cesA (cereulide synthetase gene
cluster). The screening for potential presence of
toxin related genes was conducted using Bacillus
cereus reference genes (EFSA, 2014). Results
have shown that the B. amyloliquefaciens CECT
5940 strain does not contain hemolytic enterotoxin
(nheABC), hemolysin (hblABCD), cytotoxin (cytK),
or cereulide (cesA) genes.
Following the screening for potential presence
of toxin related genes, a cytotoxicity study was
conducted to evaluate any potential production
of cyclic lipopeptides with surfactant properties.
The cytotoxicity study was conducted according
to the "Guidance on the characterization of
microorganisms used as feed additives or as
production organisms" (EFSA, 2018). In short, the
toxigenic potential of Bacillus amyloliquefaciens
CECT 5940 strain was tested on Vero C1008

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Poultry USA - January 2020

Table of Contents for the Digital Edition of Poultry USA - January 2020

Poultry USA - January 2020
Contents
Will the trade war cool off in 2020?
Poultry’s Perch: USPOULTRY invests millions in poultry industry research
US could find $1 billion poultry market in China
Feed additives that will succeed in 2020 and beyond
What role will alternative proteins play in the future?
Mar-Jac Poultry feed mill built for capacity, expansion
Poultry consumers want convenience, claims
Perdue Farms, Niman Ranch have been good for each other
Product Review
Market Place
Ad Index
Poultry USA - January 2020 - CT1
Poultry USA - January 2020 - CT2
Poultry USA - January 2020 - Poultry USA - January 2020
Poultry USA - January 2020 - Cover2
Poultry USA - January 2020 - Contents
Poultry USA - January 2020 - Will the trade war cool off in 2020?
Poultry USA - January 2020 - 3
Poultry USA - January 2020 - 4
Poultry USA - January 2020 - 5
Poultry USA - January 2020 - Poultry’s Perch: USPOULTRY invests millions in poultry industry research
Poultry USA - January 2020 - 7
Poultry USA - January 2020 - 8
Poultry USA - January 2020 - 9
Poultry USA - January 2020 - 10
Poultry USA - January 2020 - 11
Poultry USA - January 2020 - US could find $1 billion poultry market in China
Poultry USA - January 2020 - 13
Poultry USA - January 2020 - 14
Poultry USA - January 2020 - 15
Poultry USA - January 2020 - Feed additives that will succeed in 2020 and beyond
Poultry USA - January 2020 - 17
Poultry USA - January 2020 - 18
Poultry USA - January 2020 - 19
Poultry USA - January 2020 - 20
Poultry USA - January 2020 - 21
Poultry USA - January 2020 - What role will alternative proteins play in the future?
Poultry USA - January 2020 - 23
Poultry USA - January 2020 - 24
Poultry USA - January 2020 - 25
Poultry USA - January 2020 - 26
Poultry USA - January 2020 - 27
Poultry USA - January 2020 - Mar-Jac Poultry feed mill built for capacity, expansion
Poultry USA - January 2020 - 29
Poultry USA - January 2020 - 30
Poultry USA - January 2020 - 31
Poultry USA - January 2020 - 32
Poultry USA - January 2020 - 33
Poultry USA - January 2020 - 34
Poultry USA - January 2020 - 35
Poultry USA - January 2020 - 36
Poultry USA - January 2020 - 37
Poultry USA - January 2020 - 38
Poultry USA - January 2020 - 39
Poultry USA - January 2020 - Poultry consumers want convenience, claims
Poultry USA - January 2020 - 41
Poultry USA - January 2020 - 42
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Poultry USA - January 2020 - Perdue Farms, Niman Ranch have been good for each other
Poultry USA - January 2020 - 49
Poultry USA - January 2020 - 50
Poultry USA - January 2020 - Product Review
Poultry USA - January 2020 - Ad Index
Poultry USA - January 2020 - Cover3
Poultry USA - January 2020 - Cover4
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