BIFIDOBACTERIUM LACTIS BI-07 ®
TM 55-3 US 7.19
Technical Memorandum
Benefits summary
Extensive in vitro and in vivo studies
support the health-enhancing,
probiotic properties of B. lactis Bi-07.
These attributes can be summarized
as follows:
• Long history of safe use
• Well-suited to intestinal survival
- High tolerance to gastrointestinal
conditions (acid and bile)
- Strong adhesion to intestinal cell
lines
• Improves gastrointestinal health
and well-being
• Beneficial modulation of immune
functions
Introduction
A growing awareness of the
relationship between diet and health
has led to an increasing demand for
products that are able to enhance
health beyond basic nutrition. Studies
have shown that ingestion of
probiotics, or health-promoting
bacteria, is beneficial for maintaining
the microbial balance in the body.
This balance is known to particularly
enhance intestinal health and the
immune system, as well as other
physiological functions, making it a
critical factor for general human
well-being (de Moreno de LeBlanc
and LeBlanc, 2014; Kechagia et al,
2013; Vandenplas et al, 2015).
Probiotics are defined as: live
microorganisms that, when
administered in adequate amounts,
confer a health benefit on the host
(Hill et al, 2014).
Most probiotics are either lactobacilli
or bifidobacteria, although species of
other microbial genera are also
reported to have probiotic properties.
The beneficial effects of probiotics
either involve reducing risk factors for
a certain disease or improving some
of the body’s natural functions,
thereby helping to maintain the
health of the consumer. So far, these
effects have been documented
primarily in two areas, which are also
the main areas of probiotic research
for DuPont:
• gastrointestinal well-being
• beneficial modulation of the immune
system
The suggested health benefits of
probiotics are many. It should,
however, be noted that each probiotic
strain has its own specific health
benefits, and no probiotic strain elicits
all the health benefits proposed for
probiotics in general. Furthermore,
when one probiotic strain has a
certain health benefit, it cannot be
assumed that another strain, not even
of the same species, has similar
properties.
The origin of a bacterial strain, e.g.,
the human gastrointestinal tract, is no
guarantee or precondition for its
performance as a probiotic. For a
probiotic strain to be successful, it has
to fulfill certain requirements. These
will improve its functionality in the
intestine after consumption and
enhance its survival in the product.
• The strain must be safe and thus
free from potential risk-genes. This
requires identification by appropriate
molecular techniques
• The strain has to be able to resist
acid and bile
• The strain must have scientifically
proven health benefits
• The strain should have good
technological properties, such as
the ability to survive in the final
consumer product in sufficient
counts until end of shelf-life,
whether food or dietary
supplements.
The only certain way to establish the
true quality and value of a probiotic
strain is by systematic in vitro and in
vivo studies and, in particular, human
clinical trials.
Characteristics of the species
Bifidobacterium spp. are anaerobic,
Gram-positive, non-spore forming,
pleomorphic lactic acid bacteria
commonly found in the guts of
healthy humans and infants
(Arumugam et al, 2011; Mitsuoka 1996;
Scardovi et al, 1986; Turroni et al,
2009; Ventura et al, 2007).
Bifidobacteria were discovered in
1899 in the feces of breast-fed infants.
1
This was of particular interest to
scientists as these bacteria are
typically the most abundant species
present in the intestine of breastfed
infants and regarded as a primary
reason for the infants’ greater
resistance to disease. Today
bifidobacteria are broadly recognized
for their key role in the human
intestinal microbiota throughout life.
A high proportion of bifidobacteria in
the intestinal tract is considered
beneficial to health.
Selection and taxonomy
Bifidobacterium lactis was originally
described by Meile et al (Meile et al,
1997) and was recently re-classified as
B. animalis subsp. lactis (Masco et al,
2004). In the interests of simplicity,
DuPont refers to strains of this species
as B. lactis. B. lactis Bi-07 has been
genetically characterized and
properly classified as B. lactis by
independent labs using modern
genotypic methods, including 16S
rRNA gene sequencing and PCR using
species-specific primers (Ventura and
Zink, 2002). B. lactis Bi-07 is of human
origin and has been shown to grow
well in milk. B. lactis Bi-07 has been
deposited in the American Type
Culture Collection’s safe deposit as
SD5220.
Genomics
The complete genome sequence of
B. lactis Bi-07 has been published,
allowing for more stringent strain
identity confirmation among other
genetically similar B. lactis strains
(Stahl and Barrangou, 2012). The
genome sequence of strain B. lactis
Bi-07 can be found in GenBank under
the number CP003498. The genomic
variability within the B. animalis
subsp. lactis is very low and
associated only with single-nucleotide
polymorphism (SNP) or with
insertions or deletions of bases
(INDEL) (Briczinski et al, 2009; Milani
et al, 2013). However, SNPs and
Figure 1. Scanning electron micrograph of Bifidobacterium lactis Bi-07 (©DuPont)
INDELs can be applied to differentiate safety (Aguirre et al, 1994; Salminen
between B. animalis subsp. lactis et al, 1998; Borriello et al, 2003; Boyle
strains by a method described by et al, 2006). Furthermore, B. lactis has
Briczinski et al. The strain-specific been present in human food for
typing technique can be used in decades.
clinical, regulatory, and commercial
This species B. lactis is considered
settings to identify the selected strain
safe for human consumption and
(Briczinski et al, 2009). Furthermore, a
marketed as a food supplement
thorough assessment for safety
worldwide. The strain B. lactis Bi-07 is
showed no evidence of genetic risk
manufactured in accordance with U.S.
elements (Morovic et al, 2017)
Food & Drug Administration’s (FDA)
regulations. B. lactis strain Bi-07, is
Consistent strain identity
recognized as GRAS with no objection
For a strain with documented probiotic
from U.S. FDA (GRAS Notification
activity, it is very important that it is
#445, U.S. Food & Drug
not subjected to any genetic or
Administration, 2013). It is enlisted in
physiological change during
the Inventory of Microorganisms with
processing. To maintain the quality,
Documented History of Use in Human
purity, and consistency of each
Food (Bourdichon 2012) and the
production batch of the strain, DuPont
Qualified Presumption of Safety list
makes rigorous bacterial frozen seed
by EFSA (Ricci et al, 2017). No harmful
inventories to reduce the risk of
metabolic or toxigenic activities are
genetic drift over time and maintain
associated with B. lactis.
strain integrity. DuPont also performs
bacterial identification based on 16s In human clinical studies, B. lactis
rRNA gene sequence similarity for Bi-07 has been safely used as a single
every produced batch of probiotics. entity and in combination with other
probiotics and/or prebiotics. The age
Safe for consumption
of the subjects in these trials ranged
Bifidobacterium spp. has long been
between 7 months (toddlers) and
considered safe and suitable for
~70 years. Most of the studies were
human consumption with several
conducted in healthy subjects, but
published studies addressing its
2
some also involved subjects with
atopic dermatitis, functional bowel
disorders, or infections requiring
antibiotic treatment. No serious
adverse effects have been associated
with the administration of B. lactis
Bi-07.
The safety of formulas containing
probiotics and prebiotics as well as
fecal microbiology and colonization of
the probiotic was evaluated in a
multicenter, double-blind, placebo-
controlled, randomized study with
healthy toddlers, 12-34 months of age.
Toddlers were randomly divided into
three groups: control, probiotic (B.
lactis Bi-07), and synbiotic (B. lactis
Bi-07 and fructo-oligosaccharide, FOS).
B. lactis was detected significantly
more frequently in the feces of the
probiotic and synbiotic group at days 7
and 28, compared to the control group.
No significant differences were found
in fecal concentrations of Bacteroides,
Streptococcus, or total Bifidobacterium
groups. There were no statistically
significant differences across formula
groups in the numbers and kinds of
formula-related adverse effects
(Bettler et al, 2006).
To investigate the safety and
tolerability B. lactis Bi-07 in
conjunction with defined clinical
end-points, a cohort of healthy active
adults from the trial by West et al
(West et al, 2014b) was analyzed for
routine hematology and clinical
chemistry markers. Supplementation
with a combination of L. acidophilus
NCFM and B. lactis Bi-07 (5 × 109 CFU of
each strain) had no impact on these
markers (Cox et al, 2014). This data
provides evidence supporting the use
of this probiotic supplement over a
period of 5 months in healthy active
adults without obvious safety or
tolerability issues.
Antibiotic susceptibility patterns
Antibiotic susceptibility profiles are an
important means of demonstrating
the potential of an organism to be
readily inactivated by the antibiotics
used in human therapy. Antibiotic
resistance is a natural property of
microorganisms and existed before
antibiotics became used by humans.
In many cases, resistance is due to the
absence of the specific antibiotic
target or is a consequence of natural
selection. Antibiotic resistance can be
defined as the ability of some bacteria
to survive or even grow in the
presence of certain substances that
usually inhibit or kill other bacteria.
This resistance may be inherent/
intrinsic or acquired.
Inherent or intrinsic resistance
Most, if not all, strains of a certain
bacterial species are not normally
susceptible to a certain antibiotic.
The antibiotic has no effect on these
cells, being unable to kill or inhibit
the bacterium, for example because
the target for the antibiotic may
be missing.
Acquired resistance
Most strains of a bacterial species are
usually susceptible to a given
antibiotic. However, some strains may
be resistant, having adapted to
survive antibiotic exposure. Possible
explanations for this include:
• a mutation in the gene coding for
the antibiotic’s target can make an
antibiotic less efficient. This type of
antibiotic resistance is usually not
transferable.
• a resistance gene may have been
acquired from another bacterium.
Of the acquired resistances, the latter
is of most concern, as it may be
passed on to other bacteria; including
potentially pathogenic ones.
Much concern has arisen in recent
years regarding vancomycin
resistance. Vancomycin-resistant
enterococci are a leading cause of
hospital-acquired infections and are
refractory to treatment. The
transmissible nature of genetic
elements that encode vancomycin
resistance in these enterococci is an
important mechanism of
pathogenicity.
Resistance to vancomycin in certain
lactobacilli, pediococci, and
leuconostoc is due to intrinsic factors
related to the composition of their cell
wall. It is not due to any transmissible
elements (Delcour et al, 1999). B. lactis
Bi-07 is vancomycin-sensitive.
A tetracycline resistance gene
common for B. lactis is also present in
the B. lactis Bi-07 genome in a stable
genomic location suggesting unlikely
gene transferal or alteration in
phenotypic activity (Stahl and
Barrangou, 2012). To date, antibiotic
resistance transfer cases have not
been reported for Bifidobacterium
used in foods and feed. Measurements
of antibiotic sensitivity did not
demonstrate resistance for B. lactis
Bi-07 at levels exceeding any
breakpoints defined by EFSA (Morovic
et al, 2017). The expressed level of
tetracycline resistance in B. lactis Bi-07
is, however, equal to the EFSA cut-off
value (EFSA, 2012). The antibiotic
susceptibility patterns (antibiogram)
for B. lactis Bi-07 are summarized in
Table 1.
Production of biogenic amines
Histamine and tyramine are biogenic
amines that occur naturally in a wide
range of foods including fermented
products. They are formed by the
enzymes present in the raw material
or are generated by microbial
decarboxylation of amino acids. The
consumption of food containing large
amounts of these amines can induce
adverse reactions such as nausea,
headaches, rashes, and changes in
blood pressure (Ladero et al, 2010).
3
Table 1. B. lactis Bi-07 antibiotic susceptibility profiles
Antibiogram of B. lactis Bi-07 was established using ISO 10932 IDF223 method and VetMIC Lact-1 and 2 micro-dilution plates that include all
antibiotics that are recommended by The Panel on Additives and Products or Substances used in Animal Feed (FEEDAP). Recorded Minimum
Inhibitory Concentrations (MICs) are displayed in the table below. All MIC values are below or equal to the Microbial Break Points (MBPs) defined
for Bifidobacterium (EFSA Journal 2012;10(6):2740). According to the results B. lactis Bi-07 does not bear acquired antibiotic resistance (Morovic et al,
2017).

Chloramphenicol
Erythromycin
Streptomycin

Clindamycin

Vancomycin
Tetracycline
Gentamycin

Kanamycin

Ampicillin
Gm Km Sm Tc Em Cl Ch Amp Va

Max MIC μg/ml

Bifidobacterium animalis Bi-07 64 256 64 8 0.12 <0.03 2 0.25 0.5

MBP for Bifidobacterium** 64 NR*** 128 8 1 1 4 2 2

**EFSA Journal 2012;10(6):2740.

NR***: not required

In lactic acid bacteria, production of
histamine results from the catabolism
of histidine by a histidine
decarboxylase, and production of
tyramine results from the catabolism
of tyrosine by tyrosine decarboxylase.
A specific detection method for
histidine and tyrosine decarboxylase
genes has been developed internally
to DuPont based on the scientific
literature and on the most updated
genomic databases. With this method,
no histidine or tyrosine decarboxylase
gene was identified in B. lactis Bi-07
genome (Morovic et al, 2017).
Consequently, B. lactis Bi-07 is unlikely
to produce histamine or tyramine.
L/D-lactic acid production
Lactic acid is the most important
metabolic end product of fermentation
processes by lactic acid bacteria and
other microorganisms. Lactic acid
fermentation has been used for
thousands of years in the production
of fermented foods. Due to its
molecular structure, lactic acid has
two optical isomers. One is known as
L(+)-lactic acid and the other, its mirror
image, is D(-)-lactic acid.
In humans, animals, plants and that probiotics have to be consumed
microorganisms, L(+)-lactic acid is a in sufficient numbers to provide the
normal intermediate or end product of desired health benefit. It is likely that
the carbohydrate and amino acid different strains and different effects
metabolic processes whereas, D(-)- require different dosages.
lactic acid was thought to be “non-
Delivering the proper dose of B. lactis
physiological” and a possible cause for
Bi-07 over the shelf-life of a product
lactate acidosis.
can be dependent on many factors
Although some species of probiotic such as formulation, processing,
cultures, as nutritional ingredients packaging, and storage temperature
that may produce D(-)-lactic acid, have and humidity. It is important to
been safely administered to infants consider these factors and run
(Connolly et al, 2005), Codex specifies stability trials to develop sound
that only L(+)-lactic acid producing products.
cultures can be used for infant
Health-related properties
formula and follow up formula. B.
B. lactis Bi-07 has been extensively
lactis Bi-07 only produces L(+)-lactic
studied for its health sustaining or
acid (Table 2).
promoting effects. The mechanisms
Product Stability of action have been demonstrated in
Today there is a general consensus animal models and the efficacy of
Table 2. L/D Lactic acid production
L/D Lactic acid production
100/0
L/D lactic acid production
molar ratio
Boehringer Mannheim/R-Biopharm UV-method
Source: DuPont internally-generated data

4
B. lactis Bi-07 has been further
evaluated in human clinical trials.
Clinical trials to date have shown that
B. lactis Bi-07 has been included in
probiotic studies to understand its
potential role in resistance to
respiratory and gastrointestinal (GI)
infections, functional intestinal
symptoms, and the remedy of and
incidence of symptoms of antibiotic
associated diarrhea. Studies have
been conducted by using B. lactis
Bi-07 as a single strain or in
combination with other probiotic
strains and/or prebiotics. The key
findings of this research are
summarized below.
Benefits to intestinal health and
well-being
The human GI tract is an extremely
complex ecosystem and represents
the host’s greatest area of contact
with the environment. This ecosystem
comprises:
• the gastrointestinal epithelium
• immune cells
• resident microbiota
The primary function of the human GI
tract has long been considered to be
the digestion and absorption of
nutrients and the excretion of waste
end-products. In recent years,
however it has become recognized
that the GI tract fulfills many other
functions, which are essential to our
well-being. The GI tract harbors a vast
number of microbial cells (1014), which
may surpass the number of cells that
make up the human body (Sender et
al, 2016).
The intestinal microbiota is estimated
to consist of at least 1000 species,
although 95-99% of all bacteria belong
to just 10 genera. Many members of
the intestinal microbiota are beneficial,
while others are potentially
detrimental or their function not
known. A higher concentration of
Figure 2. Stability of B. lactis Bi-07 culture concentrate.
n 4°C n 25°C n 30°C
1012
1011
Log CFU/g
1010
109
0 3 6 9 12 15 18 21 24 27 30 33 36
Time (months)
Source: DuPont internally-generated data
certain genera, including Lactobacillus Resistance to acid and bile and
and Bifidobacterium, is generally survival in the intestinal passage
thought to be associated with a According to the generally accepted
healthier GI tract. definition of probiotics, a probiotic
microorganism should be viable at the
The resident microbes are involved in
time of ingestion to confer health
many metabolic processes, such as
benefit. A variety of traits are believed
the fermentation of undigested
relevant for surviving passage through
carbohydrates into short-chain fatty
the GI tract, the most important of
acids, lipid metabolism, and vitamin
which is tolerance to the highly acidic
synthesis. Another important function
conditions present in the stomach and
of the intestinal microbiota is to
to the concentrations of bile salts
stimulate the maturation of the
found in the small intestine.
immune system and provide
protection against incoming, In vitro studies have shown that B.
potentially pathogenic microbes. lactis Bi-07 is resistant to low pH
conditions and has a good survival rate
When the delicate ecological balance
in the presence of bile at the
of this highly complex microbial
concentrations present in the
community is disturbed by
duodenum (Ding and Shah, 2007;
environmental or physiological factors,
DuPont, Internally generated data)
predisposition to infectious and
(Table 3). Accordingly, B. lactis Bi-07
immuno-inflammatory diseases is
consumption elevates B. lactis counts
enhanced. It may then become
in feces during consumption (Bettler et
necessary to re-establish a beneficial
al, 2006; Childs et al, 2014; Forssten et
microbiota. Research has shown that
al, 2014; Larsen et al, 2011; Ringel-Kulka
specific probiotic strains can be used
et al, 2011) suggesting survival of
to optimize the composition and
gastrointestinal passage by the strain.
activity of the intestinal microbiota
and, thus, to reduce the risk for a range Adhesion to intestinal mucosa
of diseases or unfavorable conditions While adhesion is not a pre-requisite
(Guarino, 2013; Lin et al, 2014; for a strain to elicit probiotic
Ouwehand, 2006; Scott et al, 2015). properties, interaction with the
intestinal mucosa is considered
5
important for a number of reasons.
Binding to the intestinal mucosa may
prolong the time a probiotic strain
can reside in the intestine. This
interaction with the mucosa brings
the probiotic in close contact with the
intestinal immune system, giving it a
better opportunity to modulate the
immune response. It may also protect
against enteric pathogens by limiting
their ability to colonize the intestine.
B. lactis Bi-07 has been demonstrated
to adhere to human epithelial cell
lines (Caco-2 and HT-29; Table 4) and
to human colonic mucus (Table 5) in
vitro.
Similar results were observed in a
study that evaluated bacterial
adhesion to Caco-2 cells, either by
using radio-labelled bacteria and
counting Caco-2-bound radioactivity
(radioactive adhesion assay), or by
quantifying Caco-2-bound bacteria
with genus or species-specific primers
via real-time PCR (non-radioactive
adhesion assay). The study found that
the Caco-2 cell adhesion activity of
seven bifidobacterial strains,
belonging to 5 different species,
varied considerably. B. lactis Bi-07 was
one of the two most adhesive strains
(Candela et al, 2005).
Intestinal bacteria must make their
way through the mucosal layer before
attaching to cell surface. The
extracellular glycocalyx matrix and
mucins are made mostly of
proteoglycans that are susceptible to
enzymatic degradation. Many
pathogens such as streptococci and
staphylococci, as well as non-
pathogenic bifidobacteria, capture
plasminogen on their outer surface.
Plasminogen is a proenzyme that is
converted to plasmin by tissue type
plasminogen activator or urokinase.
The best-known function of plasmin is
the degradation of fibrin clots, but its
protease activity also extends to
proteoglycans (Lähteenmäki et al,
Table 3. Acid and bile tolerance of B. lactis Bi-07
Survival in GI tract conditions in vitro
++ 7 0-79% survival in hydrochloric acid and 1% pepsin at
Acid tolerance
pH 3.5 for 1h at 37°C
Bile salt tolerance ++ 70-79% survival in 0.3% bile salt-containing medium
++++ Excellent +++ Very good ++ Good + Fair
Source: DuPont internally-generated data
Table 4. Adherence of B. lactis Bi-07 to human intestinal cells in vitro
In vitro adhesion to intestinal epithelial cells
HT-29 ++
Caco-2 +++
++++ Excellent +++ Very good ++ Good + Fair
Source: DuPont internally-generated data
Table 5. Adherence of B. lactis Bi-07 to human colonic mucus in vitro
Adherence of B. lactis Bi-07 to human colonic mucus in vitro
Measured by fluorescence labelling of bacteria.
Adhesion rate [%] is calculated comparing the fluorescence of
3.8% ++++
the adhered bacterial cells with the fluorescence of labelled
bacteria applied.
+ poor (< 0.5) ++ fair (0.5 - 2%) +++ good (2 – 3.5%)
++++ very good (3.5-5%) +++++ excellent (> 5%)
Source: DuPont internally-generated data
2001). Candela et al showed that B. and results in more avid binding to
lactis Bi-07 uses enolase as a surface Caco-2 cells (Candela et al, 2007;
receptor for human plasminogen and Candela et al, 2008a; Candela et al,
that B. lactis Bi-07 can acquire 2008b).
plasminogen from crude extracts of
The identification of five putative
human feces (Candela et al, 2009;
plasminogen-binding proteins among
Candela et al, 2011). It was also shown
the cell wall fraction of B. lactis Bi-07,
that plasminogen bound to B. lactis
suggest that plasminogen binding to
Bi-07 can be converted into active
B. lactis is due to the concerted action
plasmin and this increases migration
of a number of proteins located on
of B. lactis Bi-07 through fibrin-matrix
the bacterial cell surface. These
6
Table 6. B. lactis Bi-07 pathogen inhibition in vitro
B. lactis Bi-07 pathogen inhibition in vitro
Salmonella Typhimurium
Staphylococcus aureus
Escherichia coli
Listeria monocytogenes
++++ Excellent +++ Very good ++ Good + Fair
findings represent a step forward in
understanding the mechanisms
involved in Bifidobacterium-host
interaction (Candela et al, 2007;
Candela et al, 2008c).
Inhibition of pathogens
The protective role of probiotic
bacteria against gastrointestinal
pathogens is highly important to
therapeutic modulation of the enteric
microbiota. Probiotics are able to
inhibit, displace, and compete with
pathogens, although these abilities are
strain-dependent. The probiotic strains’
putative mechanisms of action against
pathogenic microorganisms include
the production of inhibitory
compounds, competition with
pathogens for adhesion sites or
nutritional sources, inhibition of the
production or action of bacterial
toxins, ability to co-aggregate with
pathogens, and the stimulation of the
immune system.
In vitro inhibition is usually
investigated using an agar inhibition
assay, where soft agar containing the
pathogen is laid over colonies of
probiotic cultures, causing the
development of inhibition zones
around the colonies. This effect may
be due to the production of acids,
hydrogen peroxide, bacteriocins, and
+
+ Typhimurium, and no bifidobacteria
+++
+
Source: DuPont internally-generated data
other substances that act as antibiotic
agents and as competition for
nutrients. It should be pointed out,
however, that the extrapolation of
such results to the in vivo situation is
not straightforward. The assessment in
Table 6 is based on such an in vitro
assay.
One in vitro study investigated how
various bifidobacteria, including B.
lactis Bi-07, and adhesive
enteropathogens (Salmonella enterica
serovar Typhimurium, Yersinia
enterocolitica, and Escherichia coli
H10407) compete for adhesion to
Caco-2 cells (Candela et al, 2005). Two
competition conditions, displacement
and exclusion, were tested for each
bifidobacterium-enteropathogen pair.
In the displacement assay, the
enteropathogen was added to the
Caco-2 cell monolayer before the
addition of the bifidobacterium. In the
exclusion assay, the bifidobacteria
were added to the Caco-2 cell
monolayer before the addition of the
enteropathogen. At the end of both
competition assays, the
bifidobacterium and enteropathogen
cells bound to Caco-2 cells were
specifically quantified using real-time
PCR. All the bifidobacterial strains
showed strong displacement activity
towards these enteropathogens.
In exclusion studies, the adhesive
bifidobacterial strains excluded Y.
enterocolitica. Only one strain of B.
bifidum exerted exclusion activity
towards S. enterica serovar
strain excluded E. coli H1040.
Enteropathogens excluded none of the
bifidobacterium strains in the
exclusion assays. These results show
the ability of B. lactis Bi-07 to compete
with pathogens for epithelial
monolayer adhesion, which plays a
possible role in protecting against, or
recovery from, pathogen colonization
(Candela et al, 2005).
B. lactis Bi-07 was shown to prevent
inflammation-dependent dysbiosis on
HT-29 cell line treated with fecal slurry
collected from healthy young adults
(Centanni et al, 2014). B. lactis Bi-07
reduced the over-growth of pathogens
such as members of Fusobacteriaceae,
Bacillaceae, and Enterococcales on the
enterocyte surface. Tumor necrosis
factor (TNF)-α-induced inflammation
increased the growth of bifidobacteria
in vitro proposedly due to an adaptive
response to maintain microbiota
homeostasis. Results suggest a
beneficial effect of B. lactis Bi-07 in the
treatment of intestinal inflammation
and in the prevention of dysbiosis.
Prebiotic utilization
Prebiotics are food ingredients that are
non-digestible for humans, but are
utilized by bacteria in the intestine.
Prebiotics have been shown to
maintain balance in the gut microbiota
by promoting growth of some strains
of bifidobacteria and lactobacilli,
whereas pathogens have in general
more restricted carbohydrate
metabolism. Most commonly used
prebiotics are complex
oligosaccharides like fructo-
oligosaccharides (FOS) and galacto-
oligosaccharides (GOS).
7
B. lactis Bi-07 can utilize panose, Figure 3. The probiotic mixture containing B. lactis Bi-07 protects the fecal microbiota from
disruption by antibiotics, as indicated by the greater dissimilarity of the microbiota of the placebo
sophrose, FOS, gentobiose, and
group compared to the baseline microbiota composition (Adapted from: Engelbrektson et al, 2006).
xylo-oligosaccharides (XOS) of
110 Probiotic Placebo
different lengths as a carbon source
in vitro (Mäkeläinen et al, 2009; 100

% similar to baseline microbiota levels

Mäkeläinen et al, 2010b). The growth
rates were close to those achieved 90
with glucose. Some growth of B. lactis
80
Bi-07 was also observed with GOS,
polydextrose, lactitol, and pullulan as 70
substrates. Xylan did not induce
growth. For XOS it was later shown 60
that they enhance the growth of B.
50
lactis (not specifically Bi-07) in colon
simulator that used human feces as
40
an inoculum (Mäkeläinen et al, 2010a).
30
Antibiotic associated diarrhea and Baseline Immediately after 4 days post- 13 days post-
stabilization of microbiota during antibiotic treatment antibiotic regime antibiotic regime
antibiotic treatment
B. lactis Bi-07 was included in a
five-strain formulation investigated
for its ability to stabilize the intestinal Figure 4. The probiotic mixture containing B. lactis Bi-07 promotes the maintenance of bifidobacteria
levels in the feces of antibiotic-consuming subjects during post-treatment (*p=0.030) (Adapted from:
microbiota during and after antibiotic
Engelbrektson et al, 2006).
therapy (Engelbrektson et al, 2006). In
this human trial of patients taking 140 Probiotic Placebo
Bifidobacterium counts compared to baseline

antibiotics, the probiotic product was

found to reduce the disturbance of
the total microbiota population
(Figure 3). In addition, the probiotic
product still maintained bifidobacteria
at significantly higher levels than that
of the placebo group even two weeks
after the cessation of antibiotic
therapy (Figure 4).
B. lactis Bi-07 was evaluated in a
double-blind, placebo-controlled,
randomized human clinical study as
part of a three-strain formulation (also
including L. reuteri and L. acidophilus
NCFM). A total of 243 children aged
12-36 months were recruited. During
the 14-week intervention period, a
statistically significant reduction in the
incidence and episodic frequency of
diarrhea was recorded in the probiotic
versus the placebo group (Ruiz-
Palacios et al, 1999).
Four strains from the probiotic
mixture studied by Engelbrektson and
120
100
80
60
40
20
0
Baseline
antibiotic treatment
colleagues (2006 and 2009; mixture
including B. lactis Bi-07) were
evaluated in a large clinical trial with
503 hospitalized patients subjected to
antibiotic treatment (Ouwehand et al,
2014). The trial was designed as a
placebo-controlled dose-response
study with randomization stratified
according to age, length of antibiotic
treatment and gender. The patients
consumed the probiotic mixture
* * *
Immediately after 4 days post- 13 days post-
antibiotic regime antibiotic regime
during the antibiotic course (3 to 14
days) and for 7 days thereafter. The
incidence of antibiotic associated
diarrhea (AAD) was significantly
decreased with the higher dose. For
B. lactis Bi-07, a dose response effect
was seen in positively impacting
symptoms of AAD with daily doses of
1.70 x 1010 CFU and 4.17 x 109 CFU of a
multi-strain product applied.
Incidence of Clostridium difficile

8
associated diarrhea (CDAD) and
diarrhea-associated symptoms (fever,
abdominal pain, bloating, severity and
length of diarrhea) were also reduced
in a dose-responsive manner
(Ouwehand et al, 2014).
Forssten and co-workers analyzed
whether the resilience of the GI
microbiota during antibiotic
consumption is enhanced with
probiotics (Forssten et al, 2014). Eighty
volunteers had a 7-day course of
amoxicillin and clavulanate and were
randomized to receive a concurrent
two-strain probiotic (L. acidophilus
NCFM and B. lactis Bi-07 at 12.5 x 109
CFU/d each) or placebo
supplementation ongoing for a total
of 14 days. The microbiomes
remained essentially stable in both
groups except for an elevation of
supplemented species within the
treatment group. This was in line with
the symptom assessment during the
trial which revealed only mild GI
symptoms due to the antibiotic
treatment and no statistically
significant differences in symptom
scores between the probiotic and
placebo groups.
Irritable Bowel Syndrome, IBS
Intestinal pain is one of the
symptoms of IBS. As L. acidophilus
NCFM is known to induce analgesic
receptors in rats (Rousseaux et al,
2007), Ringel-Kulka and colleagues
analyzed the gene expression levels
of mu-opioid receptor (MOR) and
cannabinoid receptor 2 (CB2) from
human mucosal biopsies (Ringel-
Kulka et al, 2014). Twenty female
subjects consumed B. lactis Bi-07 in
combination with L. acidophilus
NCFM or only L. acidophilus NCFM
for 21 days prior to donating biopsies
during sigmoidoscopy. The L.
acidophilus NCFM alone enhanced
MOR expression whereas the
combination product had no effect.
The combination product, however,
showed a trend for more efficient
reduction in the number of days with
abdominal pain, but the study was not
sufficiently powered for evaluating
the clinical outcome.
Bowel function
Constipated women were evaluated
according to Rome III criteria before
and after a 30-day open-labelled
intervention with B. lactis Bi-07
containing fresh cheese. They
consumed 30g of fresh cheese daily
with 108 CFU of B. lactis Bi-07 per
serving in the active product. The
probiotic cheese was found to
alleviate constipation and
constipation-related symptoms more
efficiently than plain fresh cheese, but
both groups responded to treatment
(Favretto et al, 2013).
Ringel-Kulka et al studied
combination of L. acidophilus NCFM
and B. lactis Bi-07 versus placebo for
the symptoms of bloating in patients
with functional bowel disorders in a
double-blind study (Ringel-Kulka et al,
2011). The global relief of GI symptoms
and satisfaction with treatment was
not affected, but bloating symptoms
were significantly reduced in
treatment group.
D’Souza and colleagues studied the
effects of combination of L.
acidophilus NCFM and B. lactis Bi-07
versus placebo on the length of days
to resolution of bloating, abdominal
pain, and altered bowel function in
patients who undertook colonoscopy
in a double-blind trial (D’Souza et al,
2015). A total of 320 patients were
randomized post-colonoscopy to the
trial. Patients in the L. acidophilus
NCFM+B. lactis Bi-07 group had a
significantly lower number of pain
days following colonoscopy when
compared with the placebo group.
There was no difference in bloating or
return to normal bowel habit days.
Subgroup analysis revealed that
patients with pre-existing abdominal
pain benefited from probiotics in
number of pain days.
Beneficial modulation of the
immune system
The human immune system is a highly
efficient and complex system for
defending the body against foreign
infectious agents (bacteria, viruses,
and parasites) as well as from
malignant cells and other noxious
agents. An immune system that
functions optimally is an important
safeguard against infectious and
non-infectious diseases. The GI tract is
the body’s largest immune organ,
containing an estimated 80% of all
antibody-producing cells. The
intestinal microbiota represents one of
the key elements in the body’s
immune defense system (Calder et al,
2013).
The immune system of a newborn is
functionally immature. Exposure to
antigens during early life is essential to
drive the development of the gut
mucosal immune system and to
maintain immune homeostasis.
Microbial antigens derived from the
intestinal microbiota and the
environment play a crucial role in the
maturation of gut-associated lymphoid
tissue and normal resistance to
disease. Reduced microbial exposure
in Western societies has also been
associated with an increased incidence
of atopic and autoimmune disorders
(Calder et al, 2013, Versini et al, 2015).
There is a significant amount of
evidence to suggest that specific
probiotic strains are able to stimulate
and regulate several aspects of
natural and acquired immune
responses. This could either be
through stimulation of the gut
immune system or modulation of
immune cell production and function
(Lei et al, 2015).
9
Probiotic bacteria with the ability to
modulate certain immune functions
may improve the response to oral
vaccination, shorten the duration or
reduce the risk of certain types of
infection, or reduce the risk of or
alleviate the symptoms of allergy and
other immune-based conditions
(Duerkop et al, 2009, Hardy et al, 2013).
Modulation of the immune system is
an area of intense study in relation to
the DuPont™ Danisco® range of
probiotics. The goal is to understand
how each strain contributes to the
maintenance and balance of optimal
immune function. The immune
system is controlled by compounds
known as cytokines. Cytokines are
hormone-like proteins made by cells
that affect the behavior of other cells
and, thereby, play an important role in
the regulation of immune system
functions. Cytokine expression can be
modulated by specific probiotic
bacteria. However, interpreting the
health relevance of changes in
cytokine levels, both from in vivo and
human studies, remains a challenge.
Expression of cytokines and other
immune markers
In vitro assays are widely used to
define the cytokine profiles of
probiotics and, thereby, determine
their immunological effects. By
measuring the impact of probiotic
bacteria during interaction with
cytokine-expressing peripheral blood
mononuclear cells (PBMCs),
information is generated that can
help determine the ability of each
strain to contribute to balanced
immune health.
B. lactis Bi-07 was investigated in vitro
for its ability to induce the PBMCs to
secrete selected cytokines:
interleukin (IL)-10 and IL-12. The results
were compared with a strain of
Lactococcus (Lc.) lactis and a strain of
non-pathogenic E. coli. Similar to Lc.
Figure 5. Induction of IL-10 and IL-12 by B. lactis Bi-07 in PBMCs, compared with Lc. lactis and E. coli
(Adapted from: Foligne et al, 2007).
1800 IL-10 IL-12
1000
1600 900
1400 800
700
1200
600
pg/ml
pg/ml
1000
500
800
400
600
300
400
200
200 100
0 0
Bi-07 Lc. lactis E. coli Bi-07 Lc. lactis E. coli
lactis, B. lactis Bi-07 induced moderate Figure 6. Cytokine response to bacterial DNA
amounts of IL-10. However, B. lactis (cultured with PBMC) compared with the
response to LPS (Adapted from: Lammers et
Bi-07 induced higher excretion of IL-12
al, 2003).
from PBMCs (Figure 5). This is known
to shift the immune system towards a 4500
so called Th1 type of response, which 4000
plays a key role in, for example, 3500
IL-10 pg/10E6 PBMC
warding off tumors and viruses, and 3000
antagonizes allergic response (Foligne 2500
et al, 2007).
2000
Another study including B. lactis Bi-07, 1500
investigated whether bacterial DNA 1000
plays a role in the immunomodulatory 500
effects of probiotic treatment. PBMCs 0
LPS Bi-07
from healthy donors were incubated
with pure DNA from probiotic strains.
Figure 7. Percentage of protection in an
Cytokine production was analyzed in acute murine model of inflammation (TNBS)
culture supernatants. It was shown (Adapted from: Foligne et al, 2007).
that the DNA of B. lactis Bi-07
stimulated the secretion of IL-10, 50
exceeding the IL-10 levels induced by 40
lipopolysaccharide (LPS) (Figure 6)
30
(Lammers et al, 2003). These results
% protection
indicate the IL-10 inducing effect of 20
whole cells differs from that of DNA. 10
It also suggests that, even when cells
0
die and their DNA is released, they
may modulate the immune system. -10
-20
B. lactis Bi-07 has demonstrated an Bi-07 Lc. lactis E. coli
ability to modulate the immune
system in an intestinal inflammation inflammation. B. lactis Bi-07 exerted
animal model, adding on its ability to moderate but significant protection
contribute to a balanced immune from the intestinal inflammation in
system. Figure 7 demonstrates the this model, demonstrating its ability
percentage of protection from a to interact with and balance the
chemically-induced intestinal
10
intestinal mucosal immune response
(Figure 7) (Foligne et al, 2007).
Protection from experimental
Candida albicans infection
Candida yeasts are usually present in
most people but uncontrolled
overgrowth, for example due to
medication or underlying disease, can
lead to candidiasis. Candidiasis is a
fungal infection (mycosis), caused by
species of the genus Candida,
predominantly Candida albicans.
Candidiasis encompasses infections
that range from superficial, such as
oral thrush and vaginitis, to systemic
and potentially severe diseases. The
increased incidence of Candida
infections and their increasing
resistance to antifungal antibiotics
provides a strong impetus for new
research efforts to explore the use of
probiotic bacteria for the prophylaxis
and therapy of candidiasis.
B. lactis Bi-07 has been evaluated in a
Candida infection model for its
capability to protect immunodeficient
mice from lethal candidiasis (Wagner
et al, 1997). Here the strain was found
to reduce the level of Candida
colonization in all parts of the
gastrointestinal tract and significantly
reduce the incidence and severity of
candidiasis in mice. Furthermore, the
study showed higher total levels of
IgA, IgG, and IgM and an improved
specific antibody response. In
addition, B. lactis Bi-07 induced a
stronger cell-mediated immune
response against Candida. As a result,
the lethality of the candidiasis was
significantly reduced in both adult
and new-born mice.
A further study evaluated the
capacity of two B. lactis strains, B.
lactis Bi-07 and B. lactis Bb-12, to
protect two types of immunodeficient
mice from orogastric candidiasis and
systemic candidiasis of endogenous
origin (Wagner et al, 1998). In the
article, Bi-07 is called B. infantis due to species show quantitative and
an identification mistake at the time. qualitative differences in their
The authors describe the strain as “a possible biotherapeutic effects
human isolate obtained from Rhone- (Wagner et al, 1998).
Poulenc, Madison, Wisconsin” that
Another study has looked at the
indicates the strain origin and the
effect of prior colonization with
strain being Bi-07. It was seen that
probiotic bacteria on the antibody
both bifidobacteria prolonged the
responses of immunodeficient mice
survival of C. albicans-colonized adult
subject to alimentary tract
and neonatal mice. Mice diassociated
colonization with C. albicans.
with C. albicans and B. lactis Bi-07 or
Although the probiotic bacteria did
the other B. lactis strain had
not induce a vigorous antibody
significantly fewer C. albicans in their
response to their own antigens, the
stomachs and intestines compared
study demonstrated that they altered
with mice monoassociated with C.
the antibody response to C. albicans
albicans. The presence of either of the
in mice. The authors observed mixed
two B. lactis strains in the alimentary
immunomodulatory effects from the
tract reduced the incidence of
probiotic bacteria. The probiotic
disseminated candidiasis in mice. Less
strains induced antibody responses to
systemic candidiasis of endogenous
some C. albicans antigens but
origin in mice was detected in mice
inhibited antibody responses to
colonized with B. lactis Bi-07 rather
others. However, the data indicate
than the other strain of B. lactis.
that probiotic bacteria (such as L.
Immune responses were evaluated as
acidophilus NCFM and B. lactis Bi-07),
immunoglobulins in the sera of mice
which effectively prolonged the
either monoassociated with one of
survival of immunodeficient mice
the bifidobacteria or C. albicans or
colonized with C. albicans (Wagner et
diassociated with one of the
al, 1997), also strongly stimulated the
bifidobacteria and C. albicans.
production of antibodies to C. albicans
Both bifidobacteria affected the antigens. These results suggest that
production of antibodies to C. commensal bacterial flora should be
albicans, but the effects were considered an important component
different for the two mouse types and of the humoral immune system in
the two bifidobacterial strains. protecting against candidiasis. They
Despite these differences, both also demonstrate that the presence
mouse types monoassociated with B. of certain probiotic bacteria can
lactis Bi-07, but not the other B. lactis, enhance or suppress antibody
had increased serum IgG, IgA, and responses to antigens administered
IgM compared with sera from germ- via the mucosal surfaces of the
free (GF) controls. Additionally, in both alimentary tract (Wagner et al, 2000).
types of mice diassociated with C.
Modulation of the immune system
albicans and B. lactis Bi-07, the levels
in humans
of IgG, IgA, and IgM were higher
Effects of B. lactis Bi-07
compared to the GF control sera. The
supplementation with or without GOS
two bifidobacteria strains also
on immune cell activity and intestinal
suppressed weight loss associated
microbiota of elderly individuals was
with C. albicans infection. The results
evaluated in a double-blind
show that B. lactis Bi-07 can provide
randomized placebo-controlled
important protection against
cross-over trial (Maneerat et al, 2013).
candidiasis in immunodeficient mice
During the trial subjects consumed
and that different strains of the same
11
the investigational product for 21 days
with a 28-day wash-out period in
between each treatment period. All
subjects consumed all four treatments
(B. lactis Bi-07; GOS; B. lactis Bi-
07+GOS; placebo), but due to carry-
over effects only the first treatments’
samples were analyzed statistically as
if the study was designed for parallel
groups. Phagocytosis and oxidative
burst capacity of monocytes and
granulocytes, cytokine and chemokine
concentrations, composition of fecal
microbiota, and intestinal organic acid
levels were determined. It was found
that phagocytic activity of monocytes
and granulocytes was enhanced, but
intracellular reactive oxygen species
were not elevated. The results indicate
that B. lactis Bi-07 augments
phagocytosis but not inflammation
that could be harmful on homeostatic
situation (Maneerat et al, 2013).
As to effects on immune markers, XOS
influenced the cell-surface markers of
natural killer T cells and suppressed
IL-10 secretion, B. lactis Bi-07 lowered
IL-4 secretion and IgA levels while
enhancing IL-6 secretion, and the
synbiotic supplementation altered
B-cell surface markers. No significant
effects were seen on phagocytosis,
oxidative burst, or leukocyte counts.
No clear conclusions could be made
on mechanisms of immune effects of
treatments, but XOS and B. lactis Bi-07
were suggested to present an
immunostimulatory effect potentially
beneficial for subjects with suppressed
Th1 response (elderly) or excessive Th2
activity (atopic) (Childs et al, 2014).
The ability of B. lactis Bi-07 to
stimulate specific immunity has been
evaluated in a human study
measuring primary immune reaction
following vaccination. Human
volunteers were orally vaccinated
using cholera vaccine as a vaccination
model. In addition, they received
either a placebo (maltodextrin) or B.
lactis Bi-07. Supplementation with B.
lactis Bi-07 or the placebo started on
day 0 and continued for 21 days. The
subjects consumed two capsules per
day with 1010 CFU B. lactis Bi-07 or two
capsules per day with maltodextrin
(control). On day 7 and 14, the subjects
received the oral vaccine. Blood
samples were collected on day 0, 21,
and 28, and antigen-specific
antibodies IgG (immunoglobulin G)
were determined. Supplementation
with B. lactis Bi-07 showed the
tendency to increase specific IgG
induction compared to the placebo
group. This indicates the stimulation
of specific immunity by B. lactis Bi-07
(Figure 8) (Paineau et al, 2008).
Figure 8. Relative change in specific IgG
titre in orally vaccinated humans after
supplementation with B. lactis Bi-07 (Adapted
from: Paineau et al, 2008).
125 n Maltodextrin (control)
n B. lactis Bi-07
120
115
% change
110
105
100
95
90
0 21 28
days
Impact on respiratory health
The impact of a combination of B.
lactis Bi-07 and L. acidophilus NCFM
on respiratory health was investigated
in a randomized placebo controlled
study of 326 Chinese children, 3-5
years of age. Intake of two doses of L.
acidophilus NCFM+B. lactis Bi-07 daily
for 6 months significantly reduced the
incidence and duration of upper
respiratory tract infection (URTI)
symptoms and antibiotic use,
indicating improved efficacy
compared to a placebo or the single
strain alone. The effects were more
pronounced with L. acidophilus
NCFM+B. lactis Bi-07 than L.
acidophilus NCFM alone and
therefore the investigators speculated
that B. lactis Bi-07 might give
additional advantage for immune
defense against infections (Leyer et
al, 2009).
The same probiotic combination (L.
acidophilus NCFM+B. lactis Bi-07) was
tested for enhanced resilience against
URTI and GI illness among physically
active adults in a randomized
placebo-controlled intervention trial
with three parallel groups consuming
L. acidophilus NCFM+B. lactis Bi-07, B.
lactis Bl-04, or placebo (West et al,
2014b). During the wash-out period
and following the 150-day
consumption, the number of illness
episodes and duration were recorded
and ranked according to a categorical
subjective estimate. B. lactis BI-04
was associated with a significant
reduction of risk of URTI whereas the
combination product (L. acidophilus
NCFM+B. lactis Bi-07) only showed a
trend towards effectiveness. There
were too few GI illness episodes for
statistical analyses. However, no clear
effects were seen in innate immune
markers including 34 cytokines, white
cell differentials, and natural killer
(NK) cell and PBMC activity, that
would explain enhanced resilience
towards URTI (West et al, 2014a).
The beneficial effect of probiotic
combination (L. acidophilus NCFM+B.
lactis Bi-07) evidenced in the previous
study was evaluated further. PBMCs
were isolated to examine whether
probiotic consumption had affected
the frequency of circulating
regulatory T (Treg) cells (West et al,
2016). CD4+ T-cells were analyzed for
CD127loCD25+ surface-stained markers
and CD127loFoxP3+ intracellular-
stained markers to determine the
percentage of Treg-cells. Expression of
CCR9 and CXCR3 was evaluated in
12
effector/memory (CD45RO+) and naive
(CD45RO-) CD4+ T cells and Treg cells
using the surface-stained samples.
There were no significant differences
in CD4+ T-cell or Treg-cell subsets
between the L. acidophilus NCFM+B.
lactis Bi-07 group and the placebo
group. The intensity of physical
activity, on the other hand, had
significant effect on the frequency of
Treg-cells. After dividing all the
participants in the probiotics group
and the placebo group into tertiles
depending on the intensity of exercise
(low, medium, and high), the authors
observed a significant increase in the
frequency of Treg-cells in the samples
of medium-intensity group.
Combination of B. lactis Bi-07 and L.
acidophilus NCFM in healthy active
adults was not associated with
changes in the frequency of circulating
Treg-cells. However, regular physical
activity may have hidden the potential
influence of probiotic supplementation
on circulating Treg-cells (West et al,
2016).
Atopic dermatitis
In a human clinical trial, children with
atopic dermatitis (AD) (7-24 months
old) were given B. lactis Bi-07 (n=17), L.
acidophilus NCFM (n=17), or placebo
(n=16) (Gobel et al, 2010; Larsen et al,
2011). The fecal numbers of L.
acidophilus and B. lactis increased
significantly after intervention,
indicating survival of the ingested
bacteria. The administration of B.
lactis Bi-07 did not affect the
composition and diversity of the main
bacterial populations in feces.
Eosinophil cationic protein, IL-31,
interferon (IFN)-γ, IL-10, and IgE (total),
IgE (egg), IgE (milk) were measured
from plasma and calprotectin from
feces. No differences were found in
primary clinical or immunological
outcomes. In the B. lactis Bi-07 group,
levels of IFN-γ and IL-10 showed a
trend towards reduction. However,
post-hoc analysis using subjective
SCORAD (scoring atopic dermatitis)
clinical evaluation tool resulted in
significant improvement of scores in
B. lactis Bi-07 group (Gobel et al,
2010). The levels of Bifidobacterium
correlated positively, while the levels
of the Lactobacillus group correlated
negatively with improvement of
atopic eczema evaluated by the
atopic dermatitis score (SCORAD)
(Larsen et al, 2011). This correlation
was observed across the whole study
cohort and was not attributed to
probiotic intake.
Other health-related properties
Well-being and growth of children
Appropriate nutrition is particularly
important for children during acute
phases of illness to maximize energy
and fluid intake and to improve their
recovery. B. lactis Bi-07 has been
evaluated in synbiotic nutritional
supplements administered to 1 to
10-year old children (also including L.
acidophilus NCFM and FOS) in two
human clinical studies (Fisberg et al,
2002; Schrezenmeir et al, 2004). A
multicenter, open, randomized,
comparative study included acutely ill
children aged 1-6 years who required
antibiotic treatment for a bacterial
infection (Schrezenmeir et al, 2004).
The children received either a
synbiotic nutritional supplement, a
nutritional supplement without the
synbiotic components, or a fruit-
flavored drink with their medication.
Total energy intake, weight gain, and
fecal lactobacilli levels were
significantly greater in the group that
consumed the synbiotic formula. This
group also had the lowest absolute
rate of relapse or new bacterial
infections, but these differences were
not statistically significant. There were
no significant differences in fecal
bifidobacteria levels at the end of
antibiotic therapy, although levels
were higher in the synbiotic nutritional
supplement group. There were also no
significant differences among the
groups in relation to the duration of
illness or treatment. All three
supplements were generally well
tolerated. The results suggested that
the use of nutritional supplements
containing B. lactis Bi-07 is beneficial
and safe in children undergoing
antibiotic treatment, but its effect in
preventing infections remains unclear
(Schrezenmeir et al, 2004).
A further study – a double-blind,
randomized, 4-month study – was
conducted at 13 locations in Brazil,
Mexico, Portugal, and Spain (Fisberg
et al, 2002). The objective was to
evaluate the incidence and duration
of illness plus anthropometrics in
children who received a nutritional
supplement with or without
synbiotics. Children recruited for the
study were 1 to 6 years old and
underweight (as defined by a World
Health Organization/National Center
for Health Statistics chart (WHO/
NCHS)), but otherwise healthy.
Overall, the incidence of sickness,
number of sick days, and antibiotic
use were similar between the two
groups. However, the subgroup of
children aged 3 to 5 years consuming
the synbiotic formula who had at
least one episode of sickness,
experienced significantly fewer sick
days. This suggests that the formula
may help to reduce the duration of
sickness in underweight children. The
synbiotic group experienced a
significant reduction in constipation
across all ages. All subjects
experienced growth in relation to
height, weight and weight/height-
ratio with no differences in growth
between the synbiotic and control
feeding groups. Both supplements
used in the study were well tolerated,
and the overall incidence of adverse
events was very low (Fisberg et al,
2002). These studies provide further
evidence of the safe, beneficial use of
B. lactis Bi-07 in child nutrition.
13
Necrotizing enterocolitis in very low
birth weight infants
Necrotizing enterocolitis (NEC) is a
multifactorial disease that is common
among preterm infants with very low
birth weight. The pathogenesis
involves intestinal immaturity,
formula feeding, and abnormal
microbial colonization. B. lactis Bi-07
was examined in a randomized
double-blind trial where mothers of
preterm infants were supplemented
with combination of B. lactis Bi-07 and
L. acidophilus NCFM (Benor et al, 2014).
The objective was to evaluate if
maternal probiotic intake decreases
the incidence of NEC, sepsis, and
death. Probiotic or placebo
supplementation started from 1 to 3
days postpartum and was continued
until the infant was discharged home.
Infants were breastfed and received at
least 50% of their enteral nutrition
from breast milk. The incidence of NEC
decreased by 66% and the incidence
of Bell stage II NEC by 84% in those
infants whose mothers received
probiotics supplementation compared
with infants whose mothers received
placebo. Although the reduction of
NEC incidence was non-significant, it
may be clinically relevant (Benor et al,
2014). There was no statistical
difference in the incidence of sepsis or
death between groups.
Oxalate-degrading activity
In humans, accumulation of oxalic
acid can result in a number of
pathological conditions, including
hyperoxaluria, kidney stones, renal
failure, cardiomyopathy, and cardiac
conductance disorders. Intestinal
oxalate-degrading bacteria could
prevent hyperabsorption of oxalate
and thereby participate in the
maintenance of oxalate homeostasis.
B. lactis Bi-07 has been shown to
perform high oxalate-degrading
activity with 100% oxalate
degradation. The putative genes
induced during oxalate degradation in
B. lactis Bi-07 were characterized as
orthologues of oxalyl-CoA
decarboxylase (oxc) and formyl-CoA
transferase (frc) (Turroni et al, 2010).
The results support the use of B. lactis
species, such as B. lactis Bi-07, over
other Bifidobacterium as a potential
maintenance for oxalate homeostasis
Modulation of adipogenesis
Evidence implies an association
between gut microbiota and obesity.
The molecular mechanisms on how
gut microbes modulate adipogenesis
remain unidentified. Obesity is
characterised by dysregulated
endocannabinoid system (eCB) tone
in the intestine and adipose tissue. In
lean mice, activation of cannabinoid
receptors leads to increased
adipogenesis. Overactivation of the
eCB system by cannabinoid receptor
agonist leads to the leakage of the
intestinal barrier and in increase of
plasma LPS level in obese mice. In
contrast, cannabinoid receptor
antagonist improves gut barrier
function (Muccioli et al, 2010).
In a recent study, B. lactis Bi-07
together with L. acidophilus NCFM
were administered to obese mice to
evaluate the effect of probiotic
treatment on gut permeability and
activation of the eCB (Muccioli et al,
2010). Treatment with combination of
B. lactis Bi-07+L. acidophilus NCFM
reduced eCB system tone, gut
permeability, and inflammation in
obese mice. In addition, probiotic
ingestion reduced fat mass
development in obese mice. The
results suggest a mechanism that
links gut microbiota to the regulation
of adipogenesis and that probiotics
may have beneficial effects in
modulation of adipogenesis.
Uremia
Uremic rats presenting inflammation
and elevated intestinal bacterial
translocation were subjected to daily
intragastric B. lactis Bi-07 dosages of
109 CFU/ml for 4 weeks and compared
with uremic non-treated rats and sham
group rats (Wei et al, 2014). The
probiotic treatment efficiently reduced
bacterial translocation during uremia
to the level of non-uremic sham group.
Similarly, inflammatory markers (IL-6,
CRP, and TNF-α) and intestinal
permeability were restored to the
level of non-uremic rats, although
endotoxin levels did not react to
treatment. Moreover, histological
examination showed clear
improvement of morphology and
integrity. These preliminary findings on
B. lactis Bi-07 effects putatively have
value in several acute and long-term
inflammatory conditions associated
with bacterial translocation (Wei et al,
2014).
General health
Childs and colleagues analyzed in a
double-blind, randomized, cross-over
study the effects of prebiotic (XOS,
8 g daily), probiotic (B. lactis Bi-07, 109
CFU daily) and synbiotic (XOS and B.
lactis Bi-07) supplementation on gut
and immune function, blood lipids,
and mood (Childs et al, 2014). Both
treatments with XOS elevated fecal
bifidobacterial counts whereas the
probiotic supplementation only
elevated B. lactis. The microbiome
was not observed to be affected
otherwise although all three
treatments had distinct effects on
short-chain fatty acids (SCFAs). XOS
supplementation elevated plasma
HDL, which was linked to a trend of
lower cholesterol levels, and vitality
and happiness. The other
supplementations did not affect blood
lipids or self-reported moods.
14
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18
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