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Anaerobe 17 (2011) 369e374

Contents lists available at ScienceDirect

Anaerobe
journal homepage: www.elsevier.com/locate/anaerobe

Clinical Microbiology

Immunology and probiotic impact of the newborn and young children intestinal
microora
Eugenia Bezirtzoglou a, *, Elisabeth Stavropoulou b
a

Democritus University of Thrace, Faculty of Agricultural Development, Department of Food Science and Technology, Laboratory of Microbiology,
Biotechnology and Hygiene, Orestiada GR68200, Greece
b
Democritus University of Thrace, Medical School, Alexandroupolis GR68100, Greece

a r t i c l e i n f o

a b s t r a c t

Article history:
Received 11 January 2011
Received in revised form
23 March 2011
Accepted 30 March 2011
Available online 16 April 2011

Human body has developed a holistic defence system, which mission is either to recognize and destroy
the aggressive invaders or to evolve mechanisms permitting to minimize or restore the consequences of
harmful actions. The host immune system keeps the capital role to preserve the microbial intestinal
balance via the barrier effect. Specically, pathogenic invaders such as, bacteria, parasites, viruses and
other xenobiotic invaders are rejected out of the body via barriers formed by the skin, mucosa and
intestinal ora. In case physical barriers are breached, the immune system with its many components
comes into action in order to fence infection. The intestine itself is considered as an active organ due to
its abundant bacterial ora and to its large metabolic activity. The variation among different species or
even among different strains within a species reects the complexity of the genetic polymorphism which
regulates the immune system functions. Additionally factors such as, gender, particular habits, smoking,
alcohol consumption, diet, religion, age, gender, precedent infections and vaccinations must be involved.
Hormonal prole and stress seems to be associated to the integrity microbiota and inducing immune
system alterations. Which bacterial species are needed for inducing a proper barrier effect is not known,
but it is generally accepted that this barrier function can be strongly supported by providing benec
alimentary supplements called functional foods. In this vein it is stressed the fact that early intestinal
colonization with organisms such as Lactobacilli and Bidobacteria and possibly subsequent protection
from many different types of diseases. Moreover, this benec microora dominated but Bidobacteria
and Lactobacilli support the concept of their ability to modify the gut microbiota by reducing the risk of
cancer following their capacity to decrease b-glucoronidase and carcinogen levels. Because of their
benecial roles in the human gastrointestinal tract, LAB are referred to as probiotics, and efforts are
underway to employ them in modern nutrition habits with so-called functional foods. Members of
Lactobacillus and Bidobacterium genera are normal residents of the microbiota in the human gastrointestinal tract, in which they developed soon after birth. But, whether such probiotic strains derived
from the human gut should be commercially employed in the so-called functional foods is a matter of
debate between scientists and the industrial world. Within a few hours from birth the newborn develops
its normal bacterial ora. Indeed human milk frequently contains low amounts of non-pathogenic
bacteria like Streptococcus, Micrococcus, Lactobacillus, Staphylococcus, Corynebacterium and Bidobacterium. In general, bacteria start to appear in feces within a few hours after birth. Colonization by Bidobacterium occurs generally within 4 days of life. Claims have been made for positive effects of
Bidobacterium on infant growth and health. The effect of certain bacteria having a benec action on the
intestinal ecosystem is largely discussed during the last years by many authors. Bidobacterium is
reported to be a probiotic bacterium, exercising a benecial effect on the intestinal ora. An antagonism
has been reported between B. bidum and C. perfringens in the intestine of newborns delivered by
cesarian section. The aim of the probiotic approach is to repair the deciencies in the gut ora and
restore the protective effect. However, the possible ways in which the gut microbiota is being inuenced
by probiotics is yet unknown.
2011 Elsevier Ltd. All rights reserved.

Keywords:
Probiotics
Newborn
Intestinal
Microora
Immunology
Children

* Corresponding author. Tel./fax: 30 2552 41149, 30 210 9962452.


E-mail addresses: empezirt@agro.duth.gr (E. Bezirtzoglou), zabetoulini@hotmail.com (E. Stavropoulou).
1075-9964/$ e see front matter 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.anaerobe.2011.03.010

370

E. Bezirtzoglou, E. Stavropoulou / Anaerobe 17 (2011) 369e374

1. General colonization pattern


In the last decades, considerable actions have been invested in
research concerning new functional dairy foods and their impact
upon the human microbiota [1e4]. These actions focus to provide
us a valuable insight into the potential, effectiveness and limits of
functional foods [5,6].
Nowadays, scientists debate to appoint effective methods to
determine the incriminated food functionality in vitro or in vivo
through animal or human assessments [7]. However, an important
chapter to all these items seems to be the integrity of our immune
system.
Human body has developed a holistic defence system, which
mission is either to recognize and destroy the aggressive invaders
or to evolve mechanisms permitting to minimize or restore the
consequences of harmful actions. The host immune system keeps
the capital role to preserve the microbial intestinal balance via the
barrier effect [8,9]. Specically, pathogenic invaders such as
bacteria, parasites, viruses and other xenobiotic invaders are
rejected out of the body via barriers formed by the skin, mucosa
and intestinal ora. In case those physical barriers are breached, the
immune system with its many components comes into action in
order to fence infection. The human newborn devoid of bacteria
before birth is particularly prone to infection during the rst days of
life. This risk of infection is related to his insufciently developed
defence mechanisms and to exposure to a variety of micro-organisms [8,9]. A rapid and regular rise of the bacterial count was seen
during the rst week, the level of 109 CFU/g of feces being reached
after one week. In comparing the route of delivery in newborns
coming from the same environment (hospital), the means of total
bacterial counts were shown similar in both vaginally and cesarian
section delivered infants during this period. However, the breastfed infants have shown higher total counts in the rst days of life,
due to contamination by the mothers skin and milk. Indeed human
milk frequently contains low amounts of non-pathogenic bacteria
like Streptococcus, Micrococcus, Lactobacillus, Staphylococcus, diphtheroids and Bidobacterium. The rst bacteria colonizing the
intestine in newborns delivered by vaginal delivery are of maternal
origin mainly constituted from anaerobic bacteria. In caesarean
section infants, this ora is characterized by the lack of anaerobic
bacteria, which do not survive in contact with air. Only microaerophilic micro-organisms, facultative anaerobes and sporulate
forms of bacteria like Clostridium colonizes generally the caesarean
section newborn. In general, bacteria start to appear in feces within
few hours after birth. Escherichia coli, Staphylococcus and Streptococcus are isolated even in the very rst stool in large numbers.
These rst appearing micro-organisms are probably creating,
during the rst few days of life, a reduced environment favourable
to the immediate subsequent appearance of anaerobes. Anaerobic
bacteria belonging to the Bacteroides, Clostridium and Bidobacterium genera can be detected in feces within two days, sometimes at
high levels. Among them, Staphylococcus, Corynebacterium, Propionibacterium and Streptococcus were the genera most commonly
isolated from the newborn fecal ora at birth.
1.1. Intestinal immune function
The immune system possesses two functional actions which
includes the innate immune system and the acquired immune
system [10]. Both components of immunity produce their function
through an elaborate and dynamic network of cells, tissues and
organs that work together to protect the host following a series of
steps called the immune response [10].
Most cells of the immune system originate in bone marrow.
However, thymus and spleen keep as well an important role in

some immune cells processing. The immune system operates


throughout the host body. In particular, the cells of the immune
system are organised into specic structures classied as central
lymphoid tissue (bone marrow, thymus) and peripheral lymphoid
tissue (lymph nodes, spleen, mucosa-associated lymphoid tissue)
[10,11].
To respond to this challenge, lymphoid organs and tissue are
dispersed in the host body in form of clumps (lymph nodes)
accumulating reserves of leucocytes. Moreover, the leucocytes
circulate through the body between the organs and lymph nodes by
means of the lymphatic or blood vessels [12]. The leucocytes fall
into two broad categories; (a) the phagocytes, including neutrophil
polymorphs, monocytes and macrophages which form part of the
innate immune system; (b) lymphocytes by the means of B and
T lymphocytes which mediate the acquired immunity [13e16].
Lymphocytes and some mononuclear phagocytes can recirculate
between lymphoid and non-lymphoid tissues. This helps in allowing lymphocytes to be exposed to the antigens which they recognize
and is, therefore, valuable in the distribution of effector cells of the
immune response to the sites where they are needed [10].
The recirculation is a complex process depending on interactions
between the cells of the immune response and other cell types such
as endothelial cells [7,14e16]. Virgin lymphocytes move from the
primary to secondary lymphoid tissue via the blood activated
lymphocytes move from the spleen, lymph nodes and MALT into the
blood and hence to other lymphoid and non-lymphoid tissues
antigen-presenting cells such as macrophages and dendrites cells
may carry antigen back to lymphoid tissues from the periphery
[7,13]. The complex patterns of recirculation depend on the state of
activation of the lymphocytes, the adhesion molecules expressed by
endothelial cells and the presence of chemo tactic molecules which
selectively attract particular populations of lymphocytes or macrophages. This last type of immunity is based on the adaptation process
which creates immunological memories and allows even more
effective protection during future encounters. This process is the
basis of vaccination [10,17,18].
A group of proteins called complement are also part of the
immune system. The complement system is a biochemical cascade
that attacks the surfaces of foreign encounters, as a major humoral
component [10,19,20].
Moreover, recent research focuses on the importance of
communication within the acquired immune system and between
the innate and acquired systems [7,10]. This communication is
mediated directly cell to cell through production of cytokines by
binding to specic receptors adhering cellular surfacial sites [10].
Thereby, changes or alterations in growth, development or activity
of the target cell are induced [10]. Cytokines are involved in the
immune response following an immunological stimulus by
promoting synthesis of acute phase proteins, tissues proteolysis
and lipolysis and interacting with T lymphocytes for triggering the
acquired immune response. At this stage their will be a cellmediated response to the antigen as in their tour T lymphocytes
will produce cytokines to regulate the activity of the involved
immunocompetent and nally promote antibody production by
B lymphocytes [10] designated to eliminate the source of antigen.
We can then assume that the immune system is a remarkably
effective structure that incorporates specicity, inductibility and
adaptation. Any disturbance or any faulty programming of the
immune system is associated to a failure of host defence and fall
into three broad axes: immunodeciency, autoimmunity and
hypersensitivities.
The intestinal mucosa-associated lymphoid tissue contains
around 80% of cells within the immune system. This Gut-associated
Lymphoid Tissue (GALT) contains more lymphocytes than all of the
secondary lymphoid organs combined [21]. The defence system in

E. Bezirtzoglou, E. Stavropoulou / Anaerobe 17 (2011) 369e374

the gut is dependant to three components characteristics: the gut


ora, the gut mucosa and epithelium and the related immune
system.
The surface of the epithelium consists of intestinal glycoconjugates made up of mucus gel. These mucins and glycoconjugates play a key role in the barrier effect [8,9,22]. Mucins
derived from various cells through the gastrointestinal tract, as well
as from diet, may protect the mucosa cells from being attacked by
pathogenic invaders, ora constituents and may also act as a colonizing layer and source of energy.
For description of the interactions found between the host and
its microbiota, two new terms, namely GAC and MAC, were newly
brought in to use by Midtvedt [23e25]. A list of MACs (Microora
Associated Characteristics), contains several parameters e degradation of tryptic activity, conversion of cholesterol to coprostanol,
conversion of bilirubin to urobilinogen, absence of b-aspartylglycine, breakdown of mucin [23,24]. When microbes are totally
absent, as in germ-free animals, any recording made of the functions and structures studied can be classied as a GAC (Germ-free
Animal Characteristics). Antimicrobial drugs can change MAC to
GAC [22e26].
Exposure to exogenic invaders via the gut may be a key determinant in proper development of tolerance to innocuous antigens.
The intestinal microbiota contains the basic pool of developed
antigens supporting the immune system. Exposure to food antigens
and allergens seems to stimulate the immune system [25]. Delayed
maturation of the secretory immune system in children is associated with allergies [27,28]. Moreover, secretory IgA responses
correlate with the degree of environmental microbial exposure
[29,30]. It is then understandable that integrity of the immune
system provides an integral and effective immune response.
As already exposed the Gut-associated Lymphoid Tissue (GALT)
is the largest immune organ containing approximately 80% of all
antibody producing cells and producing 40 mg IgA/kg/day. The
intestine itself is considered as an active organ due to its abundant bacterial ora (1014 microbes) and to its large metabolic
activity [22e24]. The impact of the nutrient supply seems to
inuence considerably the immune function [23,27].
1.2. Food functionality and nutrition
During the last years new methodologies penetrate the scientic
community to offer us assess to this approach and more accurate
information. Assessments are made to evaluate the incriminated
food functionality in vitro or in vivo through animal or human
studies. Selective biomarkers for exploration of the integrity of the
immune system have been proposed. Specically, in vivo studies
include approaches of bacterial killing, radioactive measurement of
lymphocytes proliferation or cytokines production after exposure to
a stimulus, production of total or specic Ig by lymphocytes, cell
surface expression of molecules involved in antigen presentation
(MHC), circulating cytokines, cytokine receptors, size and
morphology of lymphoid organs, secretory IgA, intradermal delayed
hypersensibility response, pathogens found in animals organs and
nally severity of symptoms or disease caused [10].
In vitro studies are focused on the approach of adding the
nutrient in pure form directly to immune cells in culture under
a complete controlled environment [10]. All the above in vivo
studies measurements can be evaluated in this case, however, these
experimental environmental does not reect the host itself and its
particularities in items of quantity of nutrient availability and
inuence of other multifactorial parameters. Albeit these facts, we
must assume that the in vivo studies offer an ideal controlled model
for studies concerning the mechanisms of action but it is conceivable that actions observed must be extrapolated to the human

371

communities by dietary studies as some responses can apply to


signicant interindividual or interspecies variation [27]. This variation among different species or even among different strains
within a species reects the complexity of the genetic polymorphism which regulates the immune system functions.
Additionally factors such as gender, particular habits, smoking,
alcohol consumption, diet, religion, age, gender, precedent infections and vaccinations must be involved [17,27,28,31]. Hormonal
prole and stress seem to be associated to the integrity of the
microora by inducing immune system alterations [31,32].
Nutrition is a critical determinant of immune response [33].
Under breast-feeding the ora is less diversied than under bottlefeeding. However, by about the end of the second year, the fecal
ora of both groups of infants resembles that of adults [4,9,34]. In
formula-fed infants, Bacteroides dominated among the anaerobes
and high count of Enterobacteriaceae is found. It has been more
than an axiom that in breast-fed infants the intestinal ora is
dominated by Bidobacterium. The effect of human milk seemed to
be the result of Bidobacterium bidum proliferation, in contrast to
articial alimentation that seemed to favour Clostridium perfringens
implantation [4,9,34]. An antagonism between these bacteria
seems to be established in the newborn intestine, via the alimentation. The composition and properties of human milk, such as high
lactose, low casein and calcium phosphate and low buffering
capacity, seem to favour the development of Bidobacterium [4].
In developing countries hypoproteinemia following malnutrition ranks as an important cause of immune deciency. Antibodies
responses and cell-mediated immunity are strongly impaired due
to the atrophy of the thymus and the upcoming deciency of helper
T cells. However, it must not be understimated the impact of
undernutrition in developed countries in elderly, persons with
eating disorders, alcoholics, certain diseased patients, persons
undertaking inappropriate diets and nally premature newborns or
small weight newborns. In the case of newborns the immaturity of
the intestinal system seems to be the main associated factor [34], as
it is the case of germ-free animals which has not developed
strongly their intestinal system due to the lacking of antigenic
stimulation [22].
All antigens coming through the fecal oral route in the gut
become processed by the gut-associated lymphoid system and
induce a state of immunological tolerance, so-called oral tolerance
[36]. Oral tolerance is manifested by suppressed immune responsiveness to the same antigens when later administered systemically
[37]. This oral tolerance has been observed in humans, as well as in
animal species [38]. As discussed, germ-free animals showed lower
capacity to develop tolerance to dietary antigens compared to
conventional animals, while certain bacterial toxins, notably
cholera toxin and E. coli heat labile toxin, counteract oral tolerance
[39,40].
Protein energy malnutrition is associated with a signicant
impairment of cell-mediated immunity, phagocyte function,
complement system, secretory immunoglobulin A antibody
concentrations and cytokine production [33]. Micronutrients such
as zinc, selenium, iron, copper, vitamins and folic acid can reduce
immunity [33]. However, the processes of the immune system are
restored after enabling the decient nutrient [1,33].
1.3. Intestinal colonization ability associated factors
Newborns delivered by caesarean section begins life with
a bacteriologically clean state, and offer an ideal model to better
understand the installation of bacteria in the context of the
neonates hospital. In babies born by caesarean section the rst
contact with bacteria is more fortuitous. These bacteria are coming
from the environment or the hospital staff. These bacteria coming

372

E. Bezirtzoglou, E. Stavropoulou / Anaerobe 17 (2011) 369e374

Fig. 1. Mean kinetic curve of CFU/g of feces in each maternity hospital.

from the hospital environment have low colonization ability during


the rst 7 days of life. After the 7th day the bacterial counts in the
newborn intestine are dependent on the environment (Fig. 1).
Anaerobic colonization is generally delayed but Bidobacterium
retrieval and E. coli presence were similar in vaginally and
caesarean section delivered infants. It is known actually that,
Bidobacterium species tend to cluster geographically in infants;
this suggests that nosocomial sources may be more important than
maternal ones. An increase incidence of C. perfringens, is reported in
relation with the hospital environment [4,9,31,34].
Hospitalization is incriminated to change the normal microora.
Changes in intestinal microora as regards antimicrobial resistance
of the bacteria and changes of bacteria species are noted. Also, in
newborns, intestinal colonization by Klebsiella, as well as by other
Enterobacteriaceae occurs. It is much more pronounced after
caesarean section. It has been observed a delay in Bidobacterium
colonization, a predominance of Bacteroides, especially after vaginal
delivery and an increased incidence of Clostridium species. No
differences seem to exist between term and preterm infants
[4,9,31,34].
Another contributor to the immune system integrity is the
intestinal microbiota itself as bacteria like Bacteroides fragilis and
E. coli can balance the levels of vitamins in our organism [33]. Over
nutrition and obesity seems also to reduce immunity [33]. Intestinal microbiota balance is believed to contribute to the immunological integrity of the host via the barrier effect which constricts of
a drastic factor to the elimination of exogenous pathogenic bacteria
[8]. This barrier effect can be bleached by different disease states or
by alterations of the microora caused by antibiotics or other
factors [8,33,35].

1.4. Probiotic impact of the intestinal microora


Which bacterial species are needed for inducing a proper barrier
effect is not known, but it is generally accepted that this barrier
function can be strongly supported by providing benec alimentary
supplements called functional foods. In this vein it is stressed the
fact that early intestinal colonization with organisms such as Lactobacilli and Bidobacteria and possibly subsequent protection
from many different types of diseases [34].
Moreover, this benec microora dominated but Bidobacteria
and Lactobacilli support the concept of their ability to modify the
gut microbiota by reducing the risk of cancer following their
capacity to decrease b-glucoronidase and carcinogen levels [41]. In
vitro studies with Lactobacillus rhamnosus and Bidobacterium and
an in vivo study with L. rhamnosus showed a decrease in availability
of carcinogenic aatoxin in the lumen [42e44]. The most studied
functional foods that play an important role in modulating the
immune system are; probiotics, prebiotics, fatty acids and avonoids/antioxidants.

Probiotic micro-organisms are found in commercial fermented


foods, in traditional foods. More often, they are included in dairy
products and preparations. Actually, most research has been
oriented to evaluate the effect of the probiotic supplements upon
the microora development. The most discussed been lactic acid
bacteria (LAB) and Bidobacteria [4,45].
Historically, Metchnikoff links longevity of the Caucasians
people with high consumption of fermented milks. Much later,
Tissier observations report on the benecial role of certain microorganisms called Bidobacterium sp. upon the newborn intestinal
microora [46e48]. Since the early observations of Metschnikoff in
1908, lactic acid bacteria (LAB) have been shown to reduce or
eliminate potentially pathogenic micro-organisms, as well as
various toxins, mutagens and carcinogens.
In addition, LAB can modulate the innate and adaptive immune
defence mechanisms by improving the human intestinal microbiota [49]. Recent studies suggest that LAB promotes and maintain
gastrointestinal motility, prevent inammation [50], and act
protectively against cancer development by binding to mutagens,
inactivating them through the production of antioxidants, and by
enhancing the human immune system [51]. Because of their
benecial roles in the human gastrointestinal tract, LAB is often
referred to as probiotics, and efforts are underway to employ
them in modern nutrition habits with so-called functional foods.
Members of Lactobacillus and Bidobacterium genera are normal
residents of the microbiota in the human gastrointestinal tract, in
which they developed soon after birth. But, whether such probiotic
strains derived from the human gut should be commercially
employed in the so-called functional foods is a matter of debate
between scientists and the industrial world.
Within a few hours from birth the newborn develops its normal
bacterial ora [34]. Indeed human milk frequently contains low
amounts of non-pathogenic bacteria like Streptococcus, Micrococcus, Lactobacillus, Staphylococcus, Corynebacterium and Bidobacterium [46]. In general, bacteria start to appear in feces within
a few hours after birth. Colonization by Bidobacterium occurs
generally within 4 days of life [46]. Claims have been made for
positive effects of Bidobacterium on infant growth and health [46].
The effect of certain bacteria having a benec action on the intestinal ecosystem is largely discussed during the last years by many
authors [52e56]. Bidobacterium is reported to be a probiotic
bacterium, exercising a benecial effect on the intestinal ora. An
antagonism has been reported between B. bidum and
C. perfringens in the intestine of newborns delivered by cesarian
section [57]. The aim of the probiotic approach is to repair the
deciencies in the gut ora and restore the protective effect. But the
possible ways in which the gut microora is being inuenced by
probiotics is yet unknown.
It has recently been shown that Lactobacillus, another probiotic,
administered per os can stimulate macrophage activity [55]. Stimulation of macrophage activity is unlikely to affect the composition
of the gut microora, but there is evidence of inux in the intestine
of systemically produced antibody [54]. Competition for nutrients
or adhesion receptors in the gut, production of antibacterial
substances, differences in oxidoreduction potential or pH could
explain possibly this type of antagonism. In most cases, children
consume milk and dairy products more frequently than adults.
Various bacterial species used in fermentation of dairy products
often colonize these children in high numbers, specically Lactobacillus paracasei, Lactobacillus delbrueckii lactis, Lactobacillus lactis
lactis, Leuconostoc. Bidobacterium was also found sparsely in the
children microbiota (10%), as well as in the elderly (5%) [4]. The
decline in benecial Bidobacterium sp. numbers is one of the most
marked changes in the elderly gut [31]. Moreover, Bidobacterium
species diversity observed in infants and adults decreases as

E. Bezirtzoglou, E. Stavropoulou / Anaerobe 17 (2011) 369e374

Bidobacterium adolescentis and Bidobacterium longum become


most predominant. Reduced attachment of Bidobacterium sp. to
epithelial cells may be due to changes in colon structure, chemical
composition, mucus quality and abundance, and gut functionality,
which could all be induced by the ageing process [58].
Cytokine production, antibody production and NK cell population have been shown to increase with yogurt consumption [59]
in all ages. Furthermore, elders have particular dietetic habits, as
traditional food reserves a key role in their nutrition. This may
result in colonization by bacterial species used for traditional food
preparation. A role for the intestinal ora in determining the risk
for allergy development is suggested by theoretical models of the
function of the gut-associated lymphoid system and tolerance
mechanisms.
The hygiene hypothesis was stressed successfully by comparing
epidemiological data of newborns intestinal microoras coming
from two different developing and developed countries. Colonization by E. coli is delayed in Swedish infants as compared to Pakistani
infants [60]. This must be explained by the fact that newborns in
developed encounter an overall lower number of microbial antigens compared to infants in developing countries. As result,
Swedish newborns present a delayed maturation of the salivary
secretory IgA response, compared to Pakistani infants [30].
Bidobacterium was also found sparsely in the children microbiota in developed countries. However, results seems to be
controversial as in other European countries Bidobacterium
retrieval in the children microbiota seems to be strongly associated
to the effect of breast-feeding [61]. Children developing allergies
often exhibit a transient IgA deciency at mucosal sites, which
might be a sign of an overall low immune stimulation. Allergy is
produced as hypersensibility to a stressor which can be mounts
inappropriate and exaggerated immune responses. It is known that
probiotic bacteria available on the market have demonstrated
effects on different conditions, such as prevention and reduction of
intestinal tract infections by inhibiting colonization by enteric
pathogens childhood acute diarrhoea, hypocholesterolemic action,
nutritional improvement extrapolated to the prevention of cancer,
osteoporosis and allergy [62,63].
Probiotic bacteria preparations reduce the incidence of antibiotic associated diarrhoea in children [62,63] and adults consuming
prebiotic preparations presented enhanced phagocytosis by
neutrophils and monocytes [64]. Controversial results to the above
benec effects were obtained in the case children who have been
given the probiotic L. rhamnosus during their rst six months of
life and have shown increased risk of developing respiratory
allergy by four years of age, compared to a cohort of non-treated
children [65].
Animal studies showed increasing numbers in the different
markers of immune function such as, T lymphocytes, CD4 cells,
antibodies secreting cells, lymphocyte proliferation, natural killer
cell activity, Il-1, TNF and IFN-g production, secretory IgA, phagocytes activity [66,67] following oral administration of several LAB
strains. In animal studies Salmonella typhimurium seems to be
suppressed by LAB strains and repair symptoms of enterocolitis [68].
The effect of functional foods and ingredients such as probiotics,
prebiotics, selenium, dietary antioxidants (vitamins A, E, C) on the
immune function is evident. However, the optimum intake level
has not yet been established [69]. Deciencies as well as nutrient
overdose are related to disturbances of the immune function. The
recognition of medicinal mushrooms as functional foods or as
dietary supplements is fully discussed especially in the concept of
Chinese holistic medicine and modern immunology [70]. Spirulina
seems to inhibit viral replication and so strengthening cellular and
humoral immunity, so far exciting results are obtained in inhibition
of cancer and HIV-1 [35,71].

373

Recently a new type of functional food has been developed


collecting together the dynamic prole of probiotics and prebiotics.
They are called synbiotic food. Their effect seems to be extremely
benecial for the host [72].
2. Conclusion
The links between immunity and human health are still very
poorly understood. Collecting empirical data and developing
methods of integrating predictive health impact assessment and
new functional dairy foods will be on the future research agenda in
this eld. Additionally, we must consider that cross-cultural variance, neophobia and consumers attitudes of functional food
acceptance [73].
Other necessary government actions to mitigate the health
effects of the recently developed functional dairy foods may
include making more effective strategies and forecasting systems to
alert the general public and health authorities; improving the
surveillance of immunity related diseases, in order to detect
signicant changes and orient public health responses; and
providing the public with information about the behaviour and
consumption habits they can adopt to prevent some of the adverse
health effects.
Moreover, the benets from functional foods must be considered for their reasonable applicability in specic clinical cases.
Beyond the present knowledge, it is apparent that further research
must be designed to thoroughly clarify the optimal dietetic conditions regarding functional foods in order to assure safe and accurate
feeding of the consumer.
The aim of this review was to bring together a good deal of the
so numerous data on infant intestinal ora focussing in its probiotic
impact. Controlling mechanisms and host factors that inuence
bacterial succession and the effect of alimentation contribute more
to the complexity of the problem, as well as the fact that Bidobacterium sp. tend to cluster geographically.
Benet bacteria of the newborn intestine must be considered
and extensively studied for their applicability as functional foods in
treatment of specic clinical states. It is obvious, that cooperative
research in different parts of the world needed to widen our
knowledge about the global ecology of different species, colonizing
the newborn intestine and particularly of the source and mode of
transmission.
References
[1] Calder CP, Kew S. The immune system: a target for functional foods? Br J Nutr
2002;88:S165e76.
[2] Salminen S, Bouley C, Boutron-Ruault CM, Cummings HJ, Franck A, Gibson GR,
et al. Functional food science and gastrointestinal physiology and function. Br J
Nutr 1998;80:S147e71.
[3] Roberfroid BM. Prebiotics as probiotics: are they functional foods? Am J Clin
Nutr 2000;71:S1682e7.
[4] Bezirtzoglou E, Romond BM. Occurrence of Bidobacterium in the feces of
newborns delivered by cesarian section. Biol Neonate 1990;58(5):247e51.
[5] Grajek W, Olejnik A, Sip A. Probiotics, prebiotics and antioxidants as functional foods. Acta Biochem Pol 2005;52:665e71.
[6] Jacobson FM, Silverglade B, Heller RI. Functional foods: health boon or
quackery? West J Med 2000;172:8e9.
[7] McMurray ND. Cell-mediated in nutritional deciency. Prog Food Nutr Sci
1984;8:193e228.
[8] Ducluzeau R, Raibaud P. Ecologie microbienne du tube digestif. Paris: Masson;
1979.
[9] Kleessen B, Bezirtzoglou E, Matto J. Culture-based knowledge on biodiversity,
development and stability of human gastrointestinal microora. Micr Ecol
Health Dis 2000;2:S53e63.
[10] Roitt I, Brostoff J, Male D. Immunology. London, New York: Gower Medical
Publishing; 1985.
[11] Pancer Z, Cooper M. The evolution of adaptive immunity. Ann Rev Immunol
2000;24:497e518.
[12] Zen K, Parkos C. Leucocytes epithelial interactions. Curr Opin Cell Biol 2003;
15:557e64.

374

E. Bezirtzoglou, E. Stavropoulou / Anaerobe 17 (2011) 369e374

[13] Guermonprez P, Valladeau J, Zitvogel L, Thery C, Amirorena S. Antigen preparation and T cell stimulation by dendritic cells. Ann Rev Immunol 2000;20:621e67.
[14] Ryter A. Relationship between ultrastructure and specic functions of
macrophages. Comp Immunol Microbiol Infect Dis 1985;82:119e33.
[15] Kariyawasam H, Robinson D. The eosinophil: the cell and its weapons, the
cytokines, its locations. Semin Resp Crit Care Med 2006;27:117e27.
[16] Middleton D, Curan M, Maxwell L. Natural killer cells and their receptors.
Transpl Immunol 2002;10:147e64.
[17] Plotkin S. Vaccines: past, present and future. Nat Med 2005;11:S5e11.
[18] Lange T, Perras B, Fehm LH, Born J. Sleep enhances the human antibody
response to Hepatitis A vaccination. Psychosom Med 2003;65:831e5.
[19] Rus H, Cudrici C, Niculescu F. The role of the complement system in innate
immunity. Immunol Res 2005;33:103e12.
[20] Liszeski M, Farries T, Lublin D, Rooney I, Atkinson J. Control of the complement
system. Adv Immunol 2000;61:201e83.
[21] Nishimura M, Fujiyama Y, Niwakawa M, Sasaki T, Bamba T. In vivo cytokine
responses in gut-associated lymphoid tissue (GALT)and spleen following oral
administration of staphylococcal enterotoxin B. Immunol Lett 2002;81:77e85.
[22] Midtvedt T. Microora associated characteristics (MACs) and germfree animal
characteristics (GACs) in man and animals. Microecol Ther 1985;15:295e302.
[23] Midtvedt T. Microbial functional activities. In: Hansson LA, Yolken RH, editors.
Probiotics, other nutritional factors and intestinal microora. USA: LippincottRaven; 1999. p. 79e96.
[24] Midtvedt T, Carlstedt-Duke B, Hoverstad T, Midtvedt AC, Norin E, Saxerholt H.
Establishment of a biochemically active intestinal ecosystem in ex-germ free
rats. Appl Env Microbiol 1978;53:2866e71.
[25] Bezirtzoglou E, Norin E, Chen J, Midtvedt T. Inuence of roxitromycin on
mucin degradation in rats. Microecol Ther 1995;25:168e72.
[26] Midtvedt T, Frederichsen P. Inuence of antibiotics on microbial intestinal
transformation of cholesterol to coprostanol in man. Scand J Gastrenterol
1972;12:669e72.
[27] Langley-Evans CS, Carrington LJ. Diet and the developing immune system.
Lupus 2006;15:746e52.
[28] Gleeson M, Cripps WA, Clacy RL, Husband JA, Hensley JM, Leeder RS. Ontogeny
of the secretory immune system in man. Austr NZ J Med 1982;12:255e8.
[29] Bottcher FM, Jenmalm CM, Jorksten BB. Cytokine, chemokine and secretory
IgA levels in human milk in relation to atopic disease and IgA production in
infants. Ped Allergy Immunol 2003;14:35e41.
[30] Mellander L, Carlsson B, Jalil F, Sderstrm T, Hanson L. Secretory IgA antibody response against Escherichia coli antigens in infants in relation to
exposure. J Pediatr 1985;107:430e3.
[31] Bezirtzoglou E, Papadaki A, Tsiotsias A, Kotsovolou O, Konstanti M. Hormone
therapy alters the composition of the vaginal microora in ovariectomized
rats. Microb Ecol 2007;55(2008):751e9.
[32] Tsiotsias A, Voidarou C, Skoufos J, Simopoulos C, Konstandi M, Bezirtzoglou E.
Stress induced alterations in intestinal microora. Microb Ecol 2002;52:93e5.
[33] Chandra KR. Nutrition and immunology: from the clinic to cellular biology
and back again. Proc Nutr Sc 1999;58:681e3.
[34] Bezirtzoglou E. The intestinal microora during the rst weeks of life.
Anaerobe 1997;3:173e7.
[35] Kozlenko R, Henson HR. Spirulina: the immune system miracle work. Healthy
Nat J San Francisco Med Res Found 2000;3:1e4.
[36] Garside P, Mowat MA, Khoruts A. Oral tolerance in disease. Gut 1999;44:
137e42.
[37] Garside P, Mowat MA, Khoruts A. Mechanisms of oral tolerance. Cri Rev
Immunol 1997;17:119e37.
[38] Mestecky J, Huby S, Moldoveanu Z, Waldo BF, Van den Wall Bake WA,
Elson OC. Induction of tolerance in humans: effectiveness of oral and nasal
immunization routes. Ann NY Acad Sci 1996;778:194e201.
[39] Moreau CM, Corthier G. Effect of the gastrointestinal microora on induction
and maintenance of oral tolerance to ovalbumin in C3H/HeJ mice. Infect
Immun 1988;56:2766e8.
[40] Galoriau-Routhiau V, Moreau CM. Gut ora allows recovery of oral tolerance
to ovalbumine in mice after transient breakdown mediated by cholera toxin
or Escherichia coli heat-labile enterotoxin. Pediatr Res 1996;39:25e629.
[41] Haberer P, duToit M, Dicks TML, Ahrens F, Holzapfel HW. Effect of potentially
probiotic Lactobacilli on faecal enzyme activity in minipigs on a high-fat, high
cholesterol diet. Int J Food Microbiol 2003;87:287e91.
[42] Haskard C, Binnion C, Ahokas J. Factors affecting the sequestration of aatoxins by L. rhamnosus strain GG. Chem Biol Interact 2000;128:39e49.
[43] Ciegler A, Lillehoj BE, Peterson ER, Hall HH. Microbial detoxication of aatoxins. Appl Microbiol 1996;14:934e9.
[44] Oatley TJ, Rarick DM, Ji EG, Linz EJ. Binding of aatoxins B1 to Bidobacterium
in vitro. J Food Prot 2000;63:1133e6.

[45] Perdigon G, Fuller R, Raya R. Lactic acid bacteria and their effect on the
immune system. Curr Issues Intest Microbiol 2001;2:27e42.
[46] Bezirtzoglou E, Maipa V, Chotoura N, Apazidou E, Tsiotsias A, Voidarou C, et al.
Occurrence of Bidobacterium in the intestine of newborns by uorescence in
situ hybridization. Comp Immunol Microbiol Infect Dis 2006;29:345e52.
[47] Mitsuoka T, Kimura N. Die Faekal ora bei Menschen. Zbl Bakteriol Parasit
Infekt Hyg 1973;1:A226e8.
[48] Bullen IC, Tearle VP. Bidobacteria in the intestinal tract of infants: an in vitro
study. J Med Microbiol 1976;9:325e33.
[49] Drugs B, Mercenier A, Lenoir-Wijnkoop I, Arnaud C. Immunity and probiotics.
Immunol Today 1999;20:387e90.
[50] Barton EB. Innate immunity: the bridge between adaptive immunity and
inammation. Inamm Res 1999;48:232e5.
[51] Vorbach C, Harrisson R, Capecchi RM. Xanthine oxidoreductase is central to
the evolution and function of innate immunity system. Trends Immunol 2003;
24:512e7.
[52] Fuller R. Probiotics in man and animals. J Appl Bacteriol 1989;66:365e78.
[53] Parker BR. Probiotics, the other half of the antibiotics story. Anim Nutr Health
1975;29:4e8.
[54] Lilley MD, Stillwell JR. Probiotics growth promoting factors produced by
microorganisms. Science 1965;145:747e8.
[55] Perdigon G, de Macias NEM, Alvarez S, Oliver G, de Ruiz Halgado PAA. Effect of
per-orally administered lactobacilli on macrophage activation in mice. Infect
Immun 1986;53:404e10.
[56] Bezirtzoglou E, Romond BM, Romond C. Regulation of the bacterial intestinal
implantation in infant born by cesarian section. Comp Immunol Microbiol
Infect Dis 1992;15:71e4.
[57] Bezirtzoglou E, Romond BM, Romond C. Modulation of C. perfringens intestinal
colonization in infants delivered by cesarian section. Infection 1989;17:
232e6.
[58] Gibson RG, McCartney LA, Rastall AR. Prebiotics and resistance to gastrointestinal infections. Br J Nutr 2005;93:31e4.
[59] Meydani NS, Ha KW. Immunologic effects of yoghurt. Am J Clin Nutr 2002;71:
861e72.
[60] Adlerberth I, Carlsson B, de Man P, Jalil F, Khan RS, Larsson P, et al. Intestinal
colonization with Enterobacteriaceae in Pakistani and Swedish hospital
delivered infants. Acta Paediatr Scand 1991;80:602e10.
[61] Bezirtzoglou E, Romond C. Effect of the feeding practices on the establishment
of bacterial interactions in the intestine of the newborn delivered by cesarian
section. J Perinat Med 1989;17:139e43.
[62] Mattila-Sandholm T, Saarela M. In: Mattila-Sandholm T, Saarela M, editors.
Functional dairy products. Weimar, Texas: CHIPS; 1997.
[63] Bezirtzoglou E. Probiotics and the intestinal microora overtime and space. In:
Psarianos K, Kourkoutas Y, editors. Microbial implication of safe and qualitative food products. Trivandrum, Kerala, India: Research Signpost, ISBN 97881-308-0216-9; 2008.
[64] Schiffrin JE, Rochat F, Link-Auster H, Aeschlimann MJ, Donnet-Hughes A.
Immunomodulation of human blood cells following the ingestion of lactic acid
bacteria. J Dairy Sci 1995;78:491e7.
[65] Kalliomaki M, Salminen S, Poussa T, Arvilommi H, Isolauri E. Probiotics and
prevention of atopic disease:4-year follow-up of a randomized placebo
controlled trial. Lancet 2003;361:1869e71.
[66] Helm MR, Mock IN, Simpson P, Mock MD. Certain immune markers are not
good indicators of mild to moderate biotin deciency in rats. J Nutr 2001;131:
3231e6.
[67] de Luis AD, Arranz M, Aller R, Izaola O, Cuellar L, Terroba CM. Immunoenhanced enteral nutrition: effect on inammation markers in head and
neck cancer patients. Eur J Clin Nutr 2005;59:145e7.
[68] Dahiya J, Wilkie D, Vankessel A, Drew M. Potential strategies for controlling
necrotic enteritis in broiler chickens in post-antibiotic era. Anim Feed Sci
Technol 2006;129:60e88.
[69] Lopez-Varela S, Gonzalez-Gross M, Marcos A. Functional foods and the
immune system: a review. Eur J Clin Nutr 2002;56:S29e33.
[70] Hwang CL. Recent research and development of functional food in Taiwan.
J Med Invest 2007;54:89e391.
[71] Hirahashi T, Matsumoto T, Hazeki K, Saeki Y, Ui M, Seya T. Activation of the
human immune system by Spirulina. Int Immunopharmacol 2002;2:423e34.
[72] Voidarou C, Kotzampassi K, Alexopoulos A, Voudouris A, Bezirtzoglou E.
Microbial prole of the intestinal microora in ICU patients following synbiotics treatment. Intenational Congress on Microbial Ecology and Disease,
September16e18 2007, Rome, Italy.
[73] Labrecque J, Doyon M, Bellavance F, Kolodinsky J. Acceptance of functional
foods: a comparison of French, American and French Canadian consumers.
Can J Agric Econ 2006;54:647e61.