J Physiol Biochem, 65 (3), 315-328, 2009

Dietary fructooligosaccharides and potential

benefits on health
M. Sabater-Molina1, E. Larqué1, F. Torrella2 and S. Zamora1
1Department of Physiology; 2Department of Microbiology, Faculty of Biology,
University of Murcia, Spain

(Received on March, 2009)

M. SABATER-MOLINA, E. LARQUÉ, F. TORRELLA and S. ZAMORA.

Dietary fructooligosaccharides and potential benefits on health (minireview). J Phys-
iol Biochem, 65 (3), 315-328, 2009.
Fructooligosaccharides (FOS) are oligosaccharides that occur naturally in plants
such as onion, chicory, garlic, asparagus, banana, artichoke, among many others.
They are composed of linear chains of fructose units, linked by β (2-1) bonds. The
number of fructose units ranges from 2 to 60 and often terminate in a glucose unit.
Dietary FOS are not hydrolyzed by small intestinal glycosidases and reach the cecum
structurally unchanged. There, they are metabolized by the intestinal microflora to
form short-chain carboxylic acids, L -lactate, CO2, hydrogen and other metabolites.
FOS have a number of interesting properties, including a low sweetness intensity;
they are also calorie free, non-cariogenic and are considered as soluble dietary fibre.
Furthermore, FOS have important beneficial physiological effects such as low car-
cinogenicity, a prebiotic effect, improved mineral absorption and decreased levels of
serum cholesterol, triacylglycerols and phospholipids. Currently FOS are increas-
ingly included in food products and infant formulas due to their prebiotic effect stim-
ulate the growth of nonpathogenic intestinal microflora. Their consumption increas-
es fecal bolus and the frequency of depositions, while a dose of 4-15 g/day given to
healthy subjects will reduce constipation, considered one of the growing problems of
modern society, and newborns during the first months of life.

Key words: Fructooligosaccharides, Prebiotic, Infant formulas, Human milk.

Fructooligosaccharides (FOS) are genic oligosaccharides (49, 56). They are

nondigestible carbohydrates that repre-
sent one of the major classes of bifido-
Correspondence to M. Sabater-Molina (Tel.: +34 968
363942; Fax. +34 968 363963; e-mail: mariasm@um.
es).
compounds of a vegetable origin and are
found in varying concentrations in many
foods such as onions, asparagus, arti-
chokes, garlic, wheat, bananas, tomatoes
and honey (57). Their chemical structure
consists of a chain of fructose units with a
316 M. SABATER-MOLINA, E. LARQUÉ, F. TORRELLA AND S. ZAMORA
terminal glucose unit linked by β-(2-1)
glycosidic bonds, which means they can-
not be hydrolysed by human digestive
enzymes which are specific for α-glyco-
sidic bonds. The length of the chain ranges
from 2 to 60.
There are three categories of FOS, each
of which is structurally distinct: inulin,
has a polymerization degree of 2 to about
60 monomers of fructose, with an average
of 12 units (57); oligofructose is produced
by the enzymatic hydrolysis of inulin and
is defined as a fraction of oligosaccharides
with degree of polymerization lower than
20, although commercial products tend to
have a mean value of 9; these FOS are pro-
duced by the enzymatic hydrolysis of
inulin and consists of fructosyl chains of
different lengths, with glucose and fruc-
tose terminals. Finally, scFOS (short
chain fructooligosaccharides) are specifi-
cally defined as mixed chains of fructosyl
with a glucose terminal unit; they have a
maximum of 5 units and are derived from
sugar through natural fermentation
processes, producing 1-kestose (GF 2),
nistose (GF 3) and 1-fructosyl - nistose
(GF 4) in which the fructosyl units (F) are
linked at the β-(2-1) position of sucrose
(57) (Fig. 1).
Fig. 1. Chemical structure of the short chain fructooligosacharides.
J Physiol Biochem, 65 (3), 2009
In recent decades, several methods have
been described for analysing nondi-
gestible oligosaccharides (NDOS), most
of which involve gas-liquid chromatogra-
phy (21) or high performance liquid chro-
matography (HPLC) (12).
The first, HPLC method was based on
the separation of NDOS showing a degree
of polymerization of up to 15 through
low-pressure ion-exchange columns or of
impregnation columns (65, 69).
Thin layer chromatography (TLC) can
be used for identification of FOS from
sucrose, fructose and glucose (70). How-
ever, the long analyses times needed for
such methods has led to the development
of alternative methods such as the high-
performance anion-exchange chromatog-
raphy (HPAEC) which combined with
pulsed amperimetric detection (PAD) are
useful for separating a great variety of
NDOS (23, 34, 65).
Nevertheless, the identification of com-
plex mixtures of oligosaccharides contin-
ues to be difficult, and so partial charac-
terization of the specific enzymes (51),
methylation analysis (11), mass spectrom-
etry (MS), and/or nuclear magnetic reso-
nance (NMR) are used to reveal the exact
structure of FOS (65).
 FOS AND DIET
FOS are water-soluble and their sweet-
ness is 0.3-0.6 times that of sucrose,
depending on the chemical structure and
the degree of polymerization of the oligo-
saccharide (14, 70). They are highly
hygroscopic and their water holding
capacity is greater than that of sucrose and
the same as sorbitol (6, 70). The viscosity
of a FOS solution is higher than that of
sucrose at the same concentration due to
the greater molecular weight of FOS. The
enhanced viscosity of the gastrointestinal
content may delay the rate of gastric emp-
tying and the digestion and absorption of
nutrients. Their thermal stability also is
greater than that of sucrose and they. FOS
are highly stable in the normal range of
food pH (4.0-7.0) (50).
FOS can substitute sucrose as regards
many of its properties, including solubili-
ty, freezing and fusion point and crys-
talline properties (70). It has been estimat-
ed that the caloric value of FOS ranges
from 1.5 to 2.0 kcal/g, which represents
40-50% of that of digestible carbohy-
drates such as sucrose (50).
Fructooligosaccharides have interesting
properties:
• Low sweetness intensity: this proper-
ty makes them useful for various kinds of
foods where the use of sucrose is restrict-
ed due to its high sweetness (70).
• Calorie free; i.e., the human body
lacks the necessary enzymes to hydrolyze
the beta bonds, so that they are not
hydrolyzed by the digestive enzymes.
Thus, since these substances can not be
used as an energy source in the body, they
are safe for diabetics and people on slim-
ming diets (56, 70).
• Non-cariogenic, since they are not
used by Streptococcus mutans to form the
acids and insoluble β-glucans that are the
main causes of dental caries (56, 70).
J Physiol Biochem, 65 (3), 2009
317
• They behave as soluble food fibre
from a physiological point of view. They
are non-digestible carbohydrates of a veg-
etable origin that reach to the large intes-
tine, where they can be fermented by the
colonic flora to promote the growth of
bifidobacteria and prevent the growth of
potentially pathogenic microorganisms (7,
14, 36, 66). The bacterial degradation of
FOS occurs in two stages: in the first
stage, the monomers are hydrolyzed by
bacterial beta-oxidases. In the second, the
monomers released ferment anaerobically
to produce volatile fat acids (SCFA) such
as acetate, propionate and butyrate, and
gases (H2, CO2, CH4) (54).
These properties, together with their
other beneficial physiological effects
(low carcinogenicity, prebiotic effect,
improved mineral absorption, and
decreased serum cholesterol, phospho-
lipid and triacylglycerol levels) (17, 37)
defend the addition of FOS to foods as
infant formulas which, in any case, have
only very low quantities of these nutri-
ents.
Beneficial effects of FOS on the organism
1. Prebiotic effect.– It is generally
accepted that the bacterial community
resident in the human gastrointestinal
tract has a fundamental impact on intesti-
nal functioning and the human health (2,
13). This community consists of at least
1014 bacterial cells of more than 400 dif-
ferent species (8). Though the bacteria are
distributed through the gastrointestinal
tract, their diversity and numerical impor-
tance vary in the different sections (8, 60).
While the stomach and the small intestine
contain only <103 and 104-106 cells/ml,
respectively (60), the large intestine
(in particular the colon) is an intensely
populated ecosystem with a cellular densi-
318 M. SABATER-MOLINA, E. LARQUÉ, F. TORRELLA AND S. ZAMORA
ty of 1012 bacterial cells/g dry mass (2, 25).
Most of these bacteria in the colon are
strictly anaerobic (2, 13).
Generally, the bacteria of the intestinal
flora can be split into genera that are neg-
ative or beneficial for the host. Undesir-
able bacteria include species of the genera
Clostridium, Veillonella, Staphylococcus,
Proteus, and sometimes Bacteroides,
Enterococcus, Escherichia and Strepto-
coccus. These bacteria produce potential-
ly injurious substances for the host,
including toxic and carcinogenic products
(27) and they can have pathogenic effects,
such as diarrhea, infections, liver damages,
carcinogenesis and intestinal putrefaction
(24). The beneficial bacteria, which
include species of the genera Lactobacil-
lus, Bifidobacterium, Eubacterium and,
some Streptococcus, Enterococcus and
Bacteroides, are useful for aspects related
with nourishment and the disease preven-
tion (8, 27). Any increase in number and
activity of these bacterial groups is desir-
able (24).
The FOS in the diet escape enzymatic
hydrolysis in the small intestine and enter
to the cecum with their structure
unchanged. These substances are not
excreted in the faeces, which indicates that
they are thoroughly fermented in the
colon (2, 13). The use of these compo-
nents is mediated by the bacterial hydro-
lases of the colon, so that the bacteria pro-
duce glycolitic enzymes that hydrolyse
into mono- or disaccharides, which are
transported to the interior of the cell,
where they are metabolized to SCFAs, L-
lactate, CO2 and hydrogen (27, 28, 60).
These SCFAs, particularly acetate, propi-
onate, and butyrate, are the main end-
products of bacterial fermentation reac-
tions that acidify the colon (46, 65). This
decrease in the pH of the medium favours
the development of bacteria such as bifi-
J Physiol Biochem, 65 (3), 2009
dobacteria and lactobacilli, but is harmful
for the growth of potentially pathogenic
species (2, 65).
Resistance to gastric acidity, to hydrol-
ysis by human enzymes and to intestinal
absorption, as well as the fact that they are
fermented by the intestinal microflora and
selectively stimulate the growth and/or
activity of bacteria that contribute to the
welfare and health of the host, confer on
FOS their property as prebiotics. We
defined prebiotic as “a nondigestible food
ingredient that beneficially affects the host
by selectively stimulating the growth
and/or activity of one or a limited number
of bacteria in the colon, and thus improves
host health” (26).
2. Effects related to the production of
short chain fatty acids.– As mentioned
above, colonic fermentation of FOS by
bacteria produces short chain fatty acids
(SCFAs), lactate and gases as products of
their digestion. All the SCFAs are
absorbed quickly in the large intestine
where they are metabolized by the differ-
ent tissues: butyrate by the colonic epithe-
lium, propionate and acetate (in part) by
the liver, and acetate (in part) by the mus-
cle and other peripheral tissues (2, 60).
According to some authors, the
absorption of SCFAS influences the
metabolism of the host (46, 65). Acetate
and the propionate influence the carbohy-
drate and lipid metabolism (9, 57, 60).
Propionate reduces hepatic gluconeogene-
sis and inhibits urea formation in the liver
(18, 57). Acetate is a strong acid and
reduces the pH, effectively eliminating
pathogenic bacteria before these produce
metabolites that may be pre-carcinogenic
(9). Butyrate is an important source of
energy, as well as a regulator of cell
growth and differentiation in the mucous
intestinal (9).
 FOS AND DIET
3. Relief of the constipation.– Constipa-
tion is a common complaint that affects a
large part of the population, particularly
the elderly and newborns during the first
months of life. Other groups at risk are
pregnant and lactating women, weaned
children, people trying to lose weight, and
individuals with an unsettled way of life
(personnel working on shifts or the busi-
ness men that travel to long distances) (9).
The main symptoms are difficulty to defe-
cate and incomplete emptying of the rec-
tum, while the first treatment tends to be
diet-based (60).
The positive effects of a daily intake of
FOS can contribute to an increase in the
microbial mass and gas production,
increasing the cecal content of the colon
(33). The end products of FOS fermenta-
tion by the bacteria of the colon, SCFAs,
are absorbed and used by the cells of the
human colonic epithelium, stimulating
their growth as well as the absorption of
salts and water, which increases the
humidity of the cecal bolus through
osmotic pressure, resulting in increased
intestinal motility (50). This increase in
the cecal bolus stimulates its passage
through the colon, shortening its transit
time and reducing the time available for
the reabsorption of water. All these fac-
tors increase the weight of the faeces and
impart a softer composition (9).
However, despite what has been said
above, non-digestible oligosaccharides
can also have undesirable effects. Since
FOS have osmotic properties and are
thoroughly fermented in the large intes-
tine, in some subjects they may cause
intestinal discomfort and even operate as
laxatives (at high daily doses). FOS are an
important source of intestinal gas and, for
some people, rapid gas generation may
produce abdominal pain, eructation, flatu-
lence, stomach swelling, and intestinal
J Physiol Biochem, 65 (3), 2009
319
cramps. Furthermore, the high intake of
FOS can cause diarrheic processes due to
the osmotic retention of liquids in the
small and large intestine. However, the
appearance and intensity of these effects
depends on the dose, on the daily con-
sumption of the product, and can vary sig-
nificantly from one individual to another
(65). Hond et al (2000) observed that the
time of intestinal transit is not affected by
a daily dose of 4-15 g/day, although such
a dose would increase deposition frequen-
cy and fecal volume in healthy individuals
(30). In older persons suffering constipa-
tion it has been seen that the consumption
of inulin at dose of 20-40 g/day increases
deposition frequency although with great
variability between subjects and accompa-
nied in some of the cases by flatulence
(38).
4. Effects on mineral absorption.– In
several animal experimental models, the
consumption of FOS has been demon-
strated to have a positive effect on the
absorption of calcium, magnesium, iron
and zinc (54, 61, 62). As a rule, the dietet-
ic fibre binds to or sequesters the miner-
als, reducing their absorption in the small
intestine and their arrival in the colon (50.
57, 61). However, during fermentation of
the soluble fibre in the colon, the bound
or sequestered minerals are released and
become available for absorption. In addi-
tion, the high concentration of SCFAs
resulting from colonic fermentation of the
FOS reduces the pH of the colon and
increases the solubility/availability of the
minerals, particularly of calcium and mag-
nesium (57, 62). Furthermore, the FOS
may well increase the water content of the
colon, increasing at the same time the sol-
ubility of some minerals. It has been sug-
gested that SCFAs, especially butyrate
and lactate, stimulate the proliferation of
320 M. SABATER-MOLINA, E. LARQUÉ, F. TORRELLA AND S. ZAMORA
epithelial cells in the colon, increasing the
absorptive capacity of the epithelium and
improvement of gut health (59, 63).
Regular FOS consumption may be par-
ticularly recommendable for post-
menopausal women and the elderly, pre-
venting or postponing osteoporosis and
the anaemia. Increased calcium absorption
reduces the risk of osteoporosis since this
mineral increases bone density and mass
(61). Improved calcium absorption in
adolescents and post-menopausal women
has been seen to result from the consump-
tion of non-digestible oligosaccharides
(61, 63, 65).
The presence of Ca2+ and Mg2+ in the
colon may have important applications:
they can help to maintain the health of the
colon by controlling the rate of cell
turnover; high concentrations of Ca2+ in
the colonic content may help to form bile
salts or insoluble fatty acids, reducing the
harmful activity of the bile and the fatty
acids on the cells of the colon (68).
5. Regulation of lipidic metabolism.– It
has been seen that FOS reduce the level of
serum lipids, have a hypotriglyceridemic
effect, and decrease the level of serum
cholesterol, reducing the risk of diabetes
and obesity (50). Three mechanisms have
been proposed to explain these effects:
The first involves modification of the
glucose or insulin concentrations. FOS
reduce the glucose peaks in blood that
occur after eating and, as consequence, the
production of glucose and insulin induced
lipidic enzymes is largely reduced.
The second involves the production of
SCFAs in the colon. Propionate inhibits
the cholesterogenesis and lipogenesis
pathways, while acetate stimulates the
same. It seems, then, that the pattern of
FOS fermentation and, specially, the quo-
tient between acetate and propionate that
J Physiol Biochem, 65 (3), 2009
reaches the liver may act as one of the
potential lipid reducing characteristics of
FOS.
The third mechanism proposed is that
serum cholesterol is reduced due to the
precipitation and excretion of bile acids in
the intestine, which implies that the liver
uses cholesterol to synthesize bile acids.
On the other hand, changes in the con-
centration of serum cholesterol have been
related to changes in intestinal microflora.
Some strains of Lactobacillus acidophilus
assimilate the cholesterol present in the
medium, while others inhibit cholesterol
absorption through the intestinal wall
(50).
There is a high positive correlation
between the level of lipids in serum and
the incidence of cardiovascular diseases,
which makes FOS a possible tool for their
prevention (17). In rats fed a lipid-rich
diet supplemented with 100g of fruc-
tooligosaccharides/ kg of diet, a decrease
in the triglyceridemia of the animals was
observed but no protective effect against
the accumulation of hepatic triglycerides
and lypogenesis, suggesting a possible
peripheral mode of action (40). On the
other hand, obese Zucker rats fed a diet
supplemented with fructans showed a
reduction in hepatic steatosis, with no
effect on postprandial triglyceridemia
(15). This effect is probably the result of
the reduced availability of non-esterified
fatty acids in adipose tissue, since the fat
mass were reduced by the treatment. Any
protection against steatosis strongly
depends on the pattern of fermentation
(16); the high proportion of propionate
produced in the cecum (which reaches the
liver through the portal vein carries) is, at
least in animals, a key factor in the reduc-
tion of hepatic triacylglycerol synthesis
observed when oligosaccharides are given
to obese and normal Zucker rats (17).
 FOS AND DIET
Acetate can be considered a lipogenic
and cholesterogenic substrate, while pro-
pionate acts as an inhibitor of hepatic lipid
synthesis (19). The effect of a fruc-
tooligosaccharide supplemented diet (8-
20g/d) on serum lipids has been investi-
gated in numerous human studies. Three
out of eleven studies observed a signifi-
cant reduction in serum triglycerides,
while five studies observed a modest
reduction in total cholesterol and LDL -
cholesterol levels (17).
6. Influence on glycemia/insulinemia.–
Results concerning the effects of FOS on
glycemia and insulinemia are contradicto-
ry. Some data indicate that these effects
depend to a large extent on the physiolog-
ical conditions or the degree of evolution
of the disease (diabetes). It has been
observed that in rats a diet supplemented
with 10% oligofructose for 30 days
reduces postprandial glycemia and insu-
linemia by 17% and 26%, respectively.
However, the results for a glucose toler-
ance test after a night of fasting were iden-
tical in control rats and those receiving the
supplemented diet (57).
Hyperglycemia and hyperinsulinemia
may indicate diabetes mellitus, a metabol-
ic disorder in which the body is unable to
reproduce or respond to insulin. There are
two forms of diabetes, type I and II, of
which type II is the most common form.
Almost 10% of elderly people suffer from
diabetes type II, also known as the non
insulin dependence form of diabetes mel-
litus. This type of diabetes is often linked
to obesity and patients can delay or con-
trol the disease by diet. One of the objec-
tives of modifying the diet is to prevent
postprandial glucose peaks in blood (65).
In 1955, it was already known that inulin
prevented such peaks and modified the
absorption of macronutrients by delaying
J Physiol Biochem, 65 (3), 2009
321
gastric emptying and /or decreasing the
time of intestinal transit (35).
FOS also influence plasmatic glycaemia
and insulinemia through their effect on
the production of bacterial SCFAs. Propi-
onate, particularly, reduces gluconeogene-
sis and favors hepatic glycolysis. This
fatty acid also indirectly influences the
hepatic metabolism of glucose, decreasing
fatty acid concentration of plasma, a fac-
tor that it is related to gluconeogenesis
(35).
The fact that the nondigestible
oligosaccharides may influence the pro-
duction of intestinal hormones is interest-
ing, and suggests that such hormones act
as a link between the FOS fermentation
that occurs in the lower part of the intes-
tine and its systemic consequences (17).
7. Decreased risk of colon cancer.– The
diet is an important factor that influences
the prevalence of colon cancer. Diets that
contain high animal fat and protein con-
centrations and low dietary fibre concen-
trations have been associated with a
greater risk of colon cancer. Recent inves-
tigations suggest that FOS can inhibit the
process of developing colon cancer, main-
ly by the increasing the levels of beneficial
bacteria and the SCFAs produced during
the fermentation of FOS in the colon. The
bacteria that promote health inhibit the
growth of pathogenic bacteria and thus
reduce the production of the carcinogenic
substances and bacterial enzymes that
play a role in carcinogenesis in the colon.
At the same time, bacterial growth
increases biomass and the cecal bolus,
accelerating the time of colonic transit.
Consequently, the exposure time of the
colonic microbiota to potential carcino-
genic agents is reduced. There is also evi-
dence that bile salts are implicated in
colonic carcinogenesis and that FOS may
322 M. SABATER-MOLINA, E. LARQUÉ, F. TORRELLA AND S. ZAMORA
reduce their faecal concentration, proba-
bly through the reduction of colonic pH.
The presence of butyrate promotes nor-
mal cell proliferation and suppresses the
proliferation of carcinogenic cells. Fur-
thermore, it increases programmed cell
death in the transformed cells but not in
normal cells (65).
Aberrant crypts are potential precur-
sors of adenomas and carcinomas that
might develop in the colon. The adminis-
tration of oligofructosae in the diet has
been seen to meaningfully depress the
total number of aberrant crypt areas com-
pared with a control diet, suggesting that
FOS can suppress the development of
tumors in the colon (55, 67). In animals,
the administration of lyophilized B.
longum has been seen to inhibit the devel-
opment of aberrant crypts and the forma-
tion of colon and breast tumors (41;55).
The inhibition of colon cancer has been
associated with a decrease in the prolifer-
ation of cells in the colon mucosa and in
the activity of the enzyme ornithine
decarboxylase (55), a key enzyme in the
process of polyamine synthesis.
8. Modulation of the immune system.–
Recently, FOS consumption has also been
associated with beneficial effects on
immunomodulation of the intestinal
immune system. These include
immunoregulation of the intestinal secre-
tion of IgA and interferon (IFN)-γ
through the Peyer plates, the increased
expression of polymeric immunoglobulin
receptors in the small intestine of suckling
mice, and the development of the gut-
associated lymphoid tissue associated
with the intestine (GALT) (3, 57, 64).
Among the mechanism proposed for such
effects are: first, selective increase/
decrease in specific intestinal bacteria that
modulate local cytokine and antibody
J Physiol Biochem, 65 (3), 2009
productionthat the lactic acid bacteria
produced by FOS fermentation, including
their cell wall, or cytoplasm contents can
penetrate the cells of the intestinal epithe-
lium, activating GALTlymphoid tissue
associated with the intestine; the second
hypothesis suggests that the SCFAs pro-
duced by fermentation influence the
immune system through their immuno
modulator and anti-inflammatory proper-
ties (64). A third hypothesis maintains
that butyrate reduces the glutamine
requirements of the epithelial cells, leav-
ing it for the cells of the immune system
(65).
Oligosaccharides in human milk and
their role in neonates
Free oligosaccharides are natural con-
stituents of the milk of all mammals.
Quantitatively, oligosaccharides are the
third most plentiful component of human
milk after lactose and lipids (29). The
European Society for Paediatric Gas-
troenterology Hepathology and Nutri-
tion (ESPGHAN) indicates that human
milk contains approximately 70 g/l of car-
bohydrates, 90% of which is lactose and
the rest oligosaccharides (7-12 g/l) (5).
Compared with human milk, the concen-
tration of oligosaccharides in the milk of
most domesticated animals is lower by a
factor of 10 to 100 (5). The biological
functions of oligosaccharides are related
to their conformation (39). The main
components of the oligosaccharides found
in human milk are sialic acid, N - acetyl-
glucosamine, L-fucose, D-glucose and D-
galactose. These components are com-
bined in a way different to form 130 dif-
ferent oligosaccharides, such as fucosilat-
ed and neutral oligosaccharides, sialil lac-
tose and Gal (β1-4) GlcNAc (1, 59).
 FOS AND DIET
During lactation, oligosaccharide com-
position of maternal milk varies, the great-
est concentrations occurring in the first
stages (42). The highest concentrations of
oligosaccharides are found in the
colostrum (24% of total carbohydrates),
falling to 19% in the first month and to
15% in the second month (20, 45). In
addition to these changes, there is great
variability in the composition of the dif-
ferent oligosaccharides in women at the
same lactation stage (42). The oligosaccha-
ride content of human milk varies with
the gestational age of the baby, the dura-
tion of lactation, the time of day and the
genetic inheritance of the mother (44). It is
possible that these changes in the
oligosaccharide composition are due to a
programmed adjustment of the composi-
tion of the milk to cover the needs of the
children, associated with increased matu-
ration of the immune system or with the
aging of the cells responsible for milk pro-
duction (45).
1. Benefits of oligosaccharides in mater-
nal milk.– The oligosaccharides found in
maternal milk have numerous functions
that help protect the health of the breast
fed children (44). Oligosaccharides form
the soluble fibre of human milk and are
not hydrolyzed in the small intestine (20),
but reach the large intestine with their
structure intact, acting as competitive lig-
ands that protect to the child from
pathogens. They are the substrate for
colonic bacteria and thus contribute to the
differences observed in the pH and the
fecal biota between children fed human
milk and those fed infant formulations
(44).
This anti-infective characteristic of
oligosaccharides arises from their capacity
to inhibit bacterial adhesion to the epithe-
lial surface in the gastrointestinal, respira-
J Physiol Biochem, 65 (3), 2009
323
tory and urogenital tracts. Free oligosac-
charides and the glycoproteins of mater-
nal milk, both of which are present in
great quantity and variety, could prevent
the microorganism attack, acting as analo-
gous receptors that compete with the
epithelium ligands to bind to the bacteria
(42, 52). In this way, the oligosaccharides
can act in children (whose stomach pH is
not so acid as that of adults, and whose
immune system is not totally developed),
as additional protection against enteric
pathogens by inhibiting their adhesion to
the mucosa of the intestinal surface (20,
52).
Another way in which oligosaccharides
act is to decrease intestinal pH. Oligosac-
charides are a growth factor for Bifidobac-
terium bifidum var. pennylvanicus. In the
presence of lactose, this microorganism
releases SCFA and creates an acidic medi-
um that inhibits the growth of pathogenic
agents (42, 56).
Oligosaccharides may also play an
important role in the development of the
postnatal brain (10). The oligosaccharides
are hydrolyzed to monosaccharides (D-
glucose, D - galactose, N-acetylglucosa-
mine, L-fucose and sialic acid), and the
sialic acid is a basic component of the gan-
gliosides of the brain and glycoproteins.
Furthermore, the high sialic acid content
of human milk during the first lactation
week coincides with a period of rapid syn-
thesis of sialoglycoprotein and ganglio-
sides (45, 56). Galactocerebrosides are the
predominant glycolipids of myelin. The
liver is not capable of providing all the
galactose necessary during the period of
myelinization and brain development.
Another possible role of oligosaccharides
in maternal milk, of which galactose is a
principal component, would be to ensure
that galactose levels do not become a lim-
iting factor for the child in this period
(42).
324 M. SABATER-MOLINA, E. LARQUÉ, F. TORRELLA AND S. ZAMORA

Oligosaccharides are associated with reduction of pathogens, the composition

low risk of infection and diarrhea, and
with the improved development of the
immune system response and in the avail-
ability of minerals (56).
2. Possible effects of FOS addition to the
infant formulas.– The composition and
structure of the oligosaccharides of
human milk can not be accurately repro-
duced by the food industry since cow’s
milk is used as the basis. For this reason,
other groups of oligosaccharides of a veg-
etable origin (such as fructooligosaccha-
rides) are used in infant foods, in an
attempt to mimic the beneficial effects of
the oligosaccharides in human milk (56).
Few papers have studied how the dif-
ferent chemistries of the oligosaccharides
of human milk and fructooligosacarides
may affect their functions and physiologi-
cal effects on the newborn. In several clin-
ical trials it has been demonstrated that
the mixture of galactooligosaccharides
(GOS) and FOS stimulates the intestinal
flora of children fed infant formulations in
the same way as occurs in the children fed
with maternal milk (4, 5). This was seen in
the fecal bifidobacterial count (44), the
of fecal short chain fatty acids, fecal pH,
as well as in the frequency and consisten-
cy of the faeces (4, 39, 47). In several
experiments it has been observed that the
addition of FOS to the infant formulas
produces an increase in cell proliferation
in the colon mucosa, suggesting that the
consumption of FOS increases the popu-
lations of bifidobacteria and prevents
atrophy of the mucosa of the colonic
epithelium in neonates fed with a control
diet (31, 32). It was suggested that some of
the selected bacteria by FOS might
increase the production of polyamines in
the cecum (53). Polyamines namely
putrescine, spermine and spermidine, are
molecules involves in cell growth and dif-
ferenciation (43, 58). We observed that
bifidobacteria and lactobacillus were able
to produce polyamines in vitro (Fig. 2).
To elucidate this potential mechanism of
action of FOS, we evaluated the effect of
FOS consumption (8g/l) in a milk formu-
la on polyamine concentration in the cecal
content of early weaned piglets. We
showed that bifidogenic effect of FOS was
associated with an increase in bacterial
polyamine concentration in the cecal con-

Fig. 2. Polyamine concentration produced by Bifidobacterium spp., Lactobacillus fermentum and Lactobacillus
acidophilus in Falkow medium supplemented with ornithine (0.5% w/v).
Results are expressed as mean ± SEM.

J Physiol Biochem, 65 (3), 2009

 FOS AND DIET
tent of piglets of 15 days of age, but this
was mainly of putrescine with no changes
in polyamine concentration in cecal
mucose. Nevertheless, FOS consumption
decreased crypt depth and enzymatic
activities in these animals. The main
polyamines in gut mucose are spermine
and spermidine but not putrescine which
seems to have less effect in gut maturation
than the other polyamines.
Maternal milk contains a great variety
of immunological and anti-inflammatory
components. MORO et al. observed that
the consumption of a mixture of FOS
/GOS in six month old children signifi-
cantly reduced atopic dermatitis, which is
associated with delayed maturation of the
immune response at the beginning of lac-
tation and with high concentrations of
IgE due to the serum antigens obtained
from the diet (48). The consumption of
FOS has also been associated with the
decreased incidence of infections of the
respiratory tract and diarrhea in children
fed infant formulas (3).
Conclusions
In recent years, it has become accepted
that the intestinal microflora plays an
important role in the development of the
immune system of children. It has been
confirmed that the FOS are beneficial for
the health of the colon, since they stimu-
late the selective growth of bifidobacte-
rias and lactobacilli (prebiotic effect) and
hinder the attack of microorganisms
potentially pathogenic to intestinal cells.
The future selective manipulation of the
intestinal microbiota would represent a
new advance in the strategies available to
prevent allergies and infections in chil-
dren.
J Physiol Biochem, 65 (3), 2009
325
References
1. Baró, L., Jiménez, J., Martínez-Ferez, and Boza,
J.J. (2001): Bioactive compounds derived from
human milk. Ars Pharmaceutica, 42, 21-28.
2. Blaut, M. (2002): Relationship of prebiotics and
food to intestinal microflora. Eur J Nutr, 41, 11-
16,
3. Boehm, G., Jelinek, J., Knol, J., M’Rabet, L.,
Stahl, B., Vos, P. and Garssen, J. (2004): Prebi-
otics and Immune Responses. J Pediat Gastroen-
terol Nutr, 39, 772-773.
4. Boehm, G., Jelinek, J., Stahl, B., van Laere, K.,
Knol, J., Fanaro, S., Moro, G. and Vigi, V. (2004):
Prebiotics in Infant Formulas. J Clin Gastroen-
terol, 38, 76-79.
5. Boehm, G. and Stahl, B.(2007): Oligosaccharides
from Milk. J Nutr, 137, 847S-849.
6. Bornet, F.R.J. (1994): Undigestible sugars in
food products. Am J Clin Nutr, 59, 763S-769S.
7. Bornet, F.R., Brouns, F., Tashiro, Y. and Duvil-
lier, V. (2002): Nutritional aspects of short-chain
fructooligosaccharides: natural occurrence,
chemistry, physiology and health implications.
Digestive and Liver Disease, 34, 111-120.
8. Bourlioux, P., Koletzko, B., Guarner, F., and
Braesco, V. (2003): The intestine and its
microflore are partners for the protection of the
host: report on the Danone Symposium “The
intelligent intestine” Paris. 78, 675-683.
9. Brandt, L. (2001): Prebiotics enhance gut health.
Prepared Foods, 170, 7-10.
10. Carlson, S.E. (1985): N-acetylneuraminic acid
concentrations in human milk oligosaccharides
and glycoproteins during lactation. Am J Clin
Nutr, 41, 720-726.
11. Ciucanu, I. and Kerek, F. (1984): A simple and
rapid method for the permethylation of carbohy-
drates. Carbohydr Res, 131, 209-217.
12. Collins, F. and Chandorkar, K.R. Thin-layer
chromatography of fructo-oligosaccharides. J
Chromatography, 56, 167, 1971.
13. Conway, P.L. (2001): Prebiotics and human
health: the state of the art and future perspec-
tives. Scand J Nutr, 45, 13-21.
14. Crittenden, R.G. and Playne, M.J. (1996): Pro-
duction, properties and applications of food-
grade oligosaccharides. Trends in Food Science
and Technology, 7, 361.
15. Daubioul, C., De Wispelaere, L., Taper, H. and
Delzenne, N. (2000): Dietary oligofructose
lessens hepatic steatosis, but does not prevent
hypertriglyceridemia in obese Zucker rats. J
Nutr, 130, 1314-1319.
326 M. SABATER-MOLINA, E. LARQUÉ, F. TORRELLA AND S. ZAMORA
16. Daubioul, C., Rousseau, N., Demeure, R.,
Gallez, B., Taper, H., Declerck, B. and Delzenne,
N. (2002): tary fructans, but not cellulose,
decrease triglyceride accumulation in the liver of
obese Zucker fa/fa rats. J Nutr, 132, 967-073.
17. Delzenne, N.M. (2003): Oligosaccharides: state
of the art. Proc Nutr Soc, 62, 177-182.
18. Delzenne, N., Aertssens, J., Verplaetse, H., Roc-
caro, M. and Roberfroid, M. (1995): Effect of fer-
mentable fructo-oligosaccharides on mineral,
nitrogen and energy digestive balance in the rat.
Life Sciences, 57, 1579-1587.
19. Demigné, C., Rémésy, C, and Morand, C.
(1999): Short chain fatty acids. In G Gibson and
M Roberfroid, eds. Colonic Microbiota, Nutri-
tion and Health. Dordrecht, The Netherlands:
Kluwer Academic Publishers, 55-69.
20. Engfer, M.B., Stahl, B., Finke, B., Sawatzki, G.
and Daniel, H. (2000): Human milk oligosaccha-
rides are resistant to enzymatic hydrolysis in the
upper gastrointestinal tract. Am J Clin Nutr, 71,
1589-1596.
21. Englyst, H.N., Quigley, M.E. and Hudson, G.J.
(1994): Determination of dietary fiber as non-
starch polysaccharides with gas-liquid chro-
matographic, high-performance liquid chro-
matographic or spectrophotometric measure-
ment of constituent sugars. Analyst, 119, 1497-
1509.
22. Franck, A. (2002): Technological functionality of
inulin and oligofructose. Br J Nutr, 87, 287-291.
23. Gelders, G.G, Bijnens, L., Loosveld, A.M.,
Vidts, A. and Delcour, J.A. (2003): Fractionation
of starch hydrolysates into dextrins with narrow
molecular mass distribution and their detection
by high-performance anion-exchange chro-
matography with pulsed amperometric detec-
tion. J Chromatogr A, 992, 75-83.
24. Gibson, G.R. (1998): Dietary modulation of the
human gut microflora using prebiotics. Brit J
Nut, 80, 209-212.
25. Gibson, G.R. (1999): Dietary Modulation of the
Human Gut Microflora Using the Prebiotics
Oligofructose and Inulin. J Nutr, 129, 1438.
26. Gibson, G.R., Probert, H.M., Van Loo, J.A.E.
and Roberfroid, M.B. (2004): Dietary modula-
tion of the human colonic microbiota: Updating
the concept of prebiotics. Nutr Res Rev, 17, 259.
27. Gibson, G.R. and Roberfroid, M.B. (1995):
Dietary modulation of the human colonic micro-
biota: introducing the concept of prebiotics. J
Nutr, 125, 1401-1412.
28. Grizar, D. and Barthomeuf, C. (1999): Non-
digestible oligosaccharides used as prebiotic
agents: mode of production and beneficial effects
J Physiol Biochem, 65 (3), 2009
on animal and human health. Reprod Nutr Dev,
39, 563-588.
29. Gudiel-Urbano, M. and Goñi, I. (2001): Human
milk oligosaccharides. The rule in the health and
development of the infants. Arch Latinoam Nutr,
51, 332-339.
30. Hond, E.D., Geypens, B. and Ghoos, Y. (2000):
Effect of high performance chicory inulin on
constipation. Nutrition Research, 20, 731-736.
31. Howard, M.D., Gordon, D.T., Garleb, K.A. and
Kerley, M.S. (1995): Dietary Fructooligosaccha-
ride, Xylooligosaccharide and Gum Arabic Have
Variable Effects on Cecal and Colonic Microbio-
ta and Epithelial Cell Proliferation in Mice and
Rats. J Nutr, 125, 2604-2609.
32. Howard, M.D, Gordon, L., Pace, K., Garleb, and
Kerley, M. (1995): Effects of dietary supplemen-
tation with fructooligosaccharides on colonic
microbiota populations and epithelial cell prolif-
eration in neonatal pigs. J Pediatric Gastroenterol
Nutr, 21, 297-303.
33. Jenkins, D.J.A., Kendall, C.W.C. and Vuksan, V.
(1999): Inulin, Oligofructose and Intestinal
Function. J Nutr, 129, 1431.
34. Kabel, M.A., Schols, H.A. and Voragen, A.G.J.
(2002): Complex xylo-oligosaccharides identi-
fied from hydrothermally treated Eucalyptus
wood and brewery’s spent grain. Carbohydr
Polym, 50, 191-200.
35. Kaur, N. and Gupta, A.K. (2002): Applications
of inulin and oligofructose in health and nutri-
tion. J Biosci, 27, 703-714.
36. Kelly, G. (2009): Inulin-Type Prebiotics –
A Review: Part 1. Altern Med Rev, 13, 315-329.
37. Kelly, G. (2009): Inulin-Type Prebiotics:
A Review (Part 2). Altern Med Rev, 14, 36-55.
38. Kleessen, B., Sykura, B., Zunft, H.J. and Blaut,
M. (1997): Effects of inulin and lactose on fecal
microflora, microbial activity, and bowel habit in
elderly constipated persons. Am J Clin Nutr, 65,
1397-1402.
39. Knol, J., Scholtens, P., Kafka, C., Steenbakkers,
J., Gro, S., Helm, K., Klarczyk, M., Schopfer, H.,
Bockler, H.M. and Wells, J. (2005): Colon
Microflora in Infants Fed Formula with Galacto-
and Fructo-Oligosaccharides: More Like Breast-
Fed Infants. J Pediatric Gastroenterol Nutr, 40,
36-42.
40. Kok, N., Taper, H. and Delzenne, N. (1998):
Oligofructose modulates lipid metabolism alter-
ations induced by a fat-rich diet in rats. J App
Toxicol, 18, 47-53.
41. Kulkarni, N. and Reddy, B.S. (1994): Inhibitory
effect of Bifidobacterium longum cultures on the
azoxymethane-induced aberrant crypt foci for-
 FOS AND DIET
mation and fecal bacterial beta-glucuronidase.
Proc Soc Exp Biol Med, 207, 278-283.
42. Kunz, C., Rudloff, S., Baier, W., Klein, N. and
Strobel, S. (2000): Oligosaccharides in human
milk: Structural, Functional, and Metabolic
Aspects. Ann Rev Nut, 20, 699-722.
43. Larqué, E., Sabater Molina, M. and Zamora, S.
(2007): Biological significance of dietary
polyamines. Nutrition, 23, 87-95.
44. McVeagh, P. and Miller, J.B. (1997): Human
milk oligosaccharides: only the breast. J Paediatr
Child Health, 33, 281-286.
45. Miller, J.B., Bull, S., Miller, J. and McVeagh, P.
(1994): The oligosaccharide composition of
human milk: temporal and individual variations
in monosaccharide components. J Pediatric Gas-
troenterol Nut, 19, 371-376.
46. Miller, T. and Wolin, L. (1996): Pathways of
acetate, propionate, and butyrate formation by
the human fecal microbial flora. Appl Environ
Microbiol, 62, 1589-1592.
47. Moro, G., Minoli, I., Mosca, M., Fanaro, S.,
elinek, J., Stahl, B. and Boehm, G. (2002):
Dosage-Related Bifidogenic Effects of Galacto-
and Fructooligosaccharides in Formula-Fed
Term Infants. J Pediatric Gastroenterology and
Nutrition, 34, 291-295.
48. Moro, G., Arslanoglu, S., Stahl, B., Jelinek, J.,
Wahn, U. and Boehm, G. (2006): A mixture of
prebiotic oligosaccharides reduces the incidence
of atopic dermatitis during the first six months of
age. Arch Dis Child, 91, 814-819.
49. Murphy, O. (2001): Non-polyol low-digestible
carbohydrates: food applications and functional
benefits. Brit J Nutr, 1, 53.
50. Mussatto, S.I. and Mancilha, I.M. (2007): Non-
digestible oligosaccharides:A review. Carbohy-
drate Polymers, 68, 597.
51. Mutter, M., Renard, C.B.G., Schols, H.A. and
Voragen, A.G.J. (1998): Mode of action of RG-
hydrolase and RG-lyase toward rhamnogalac-
turonan oligomers. Characterization of degrada-
tion products using RG-rhamnohydrolase and
RG-galacturonohydrolase. Carbohydrate
Research, 311, 155-164.
52. Newburg, D.S. (1997): Do the binding properties
of oligosaccharides in milk protect human
infants from gastrointestinal bacteria? J Nutr,
127, 980S.
53. Noack, J., Dongowski, G., Hartmann, L. and
Blaut, M. (2000): The Human Gut Bacteria Bac-
teroides thetaiotaomicron and Fusobacterium
varium Produce Putrescine and Spermidine in
Cecum of Pectin-Fed Gnotobiotic Rats. J Nutr,
130, 1225-1231,.
J Physiol Biochem, 65 (3), 2009
327
54. Ohta, A., Ohtsuki, M., Baba, S., Adachi, T.,
Sakata, T. and Sakaguchi, E. (1995): Calcium and
Magnesium Absorption from the Colon and
Rectum Are Increased in Rats Fed Fruc-
tooligosaccharides. J Nutr, 125, 2417-2424.
55. Reddy, B.S. (1999): Possible Mechanisms by
Which Pro- and Prebiotics Influence Colon Car-
cinogenesis and Tumor Growth. J Nutr, 129,
1478.
56. Rivero-Urgell, M. and Santamaria-Orleans, A.
(2001): Oligosaccharides: application in infant
food. Early Human Development, 65, S43-S52.
57. Roberfroid, M. B. and Delzenne, N. M. (1998):
Dietary fructans. Ann Rev Nutr, 18, 117-143.
58. Sabater-Molina, M., Larqué, E., Torrella, F.,
Plaza, F., Lozano, M.T., Muñoz, A. and Zamora,
S. (2009): Effects of dietary polyamines at physi-
ological doses in early weaned piglets. Nutrition.
(in press).
59. Sabharwal, H., Sjöblad, S. and Lundblad, A.
(1991): Affinity chromatographic identification
and quantitation of blood group A-active
oligosaccharides in human milk and feces of
breast-fed infants. J Pediatric Gastroenterol Nut,
12, 474-479.
60. Salminen, S., Bouley, C., Boutron-Ruault, M.C.,
Cummings, J.H., Franck, A., Gibson, G.R., Iso-
lauri, E., Moreau, M.C., Roberfroid, M. and
Rowland, I. (1998): Functional food science and
gastrointestinal physiology and function. Brit J
Nut, 80, 147-171.
61. Scholz-Ahrens, K.E., Ade, P., Marten, B.,
Weber, P., Timm, W.,il, Y., Gluer, C.C. and
Schrezenmeir, J. (2007): Prebiotics, Probiotics,
and Synbiotics Affect Mineral Absorption, Bone
Mineral Content, and Bone Structure. J Nutr,
137, 838S-8846.
62. Scholz-Ahrens, K.E. and Schrezenmeir, J. (2007):
Inulin and Oligofructose and Mineral Metabo-
lism: The Evidence from Animal Trials. J Nutr,
137, 2513S-2523.
63. Scholz-Ahrens, K., Schaafsma, G., Van der
Heuvel, E. and Schrezenmeir, J. (2001): Efffects
of prebiotics on mineral metabolism. Am J Clin
Nutr, 73, 459S-464S.
64. Seifert, S. and Watzl, B. (2007): Inulin and
Oligofructose: Review of Experimental Data on
Immune Modulation. J Nutr, 137, 2563S-2567.
65. Swennen, K., ourtin, K.M. and elcour, J.A.
(2006): Non-digestible Oligosaccharides with
Prebiotic Properties. Critical Reviews in Food
Science and Nutrition, 46, 471.
66. Trowell H. Definition of dietary fiber and
hypotheses that it is a protective factor in certain
diseases. Am J Clin Nutr, 29, 417-427, 1976.
328 M. SABATER-MOLINA, E. LARQUÉ, F. TORRELLA AND S. ZAMORA
67. Wargovich, M.J., Chen, C.D., Jimenez, A.,
Steele, V.E., Velasco, M., Stephens, L.C., Price,
R., Gray, K., and Kelloff, G.J. (1996): Aberrant
crypts as a biomarker for colon cancer: evalua-
tion of potential chemopreventive agents in the
rat. Cancer Epidemiol Biomarkers Prev, 5, 355-
360.
68. Wargowich, M.J., Eng, V.W.S. and Newmark,
H. (1984): Ca inhibits the damaging and com-
pensatory proliferating effect of fatty acids on
J Physiol Biochem, 65 (3), 2009
mouse colon epithelium. Cancer Lett, 23, 253-
258.
69. White, C.A., Corran, P.H. and Kennedy, J.F.
(1980): Analysis of underivatised D-gluco-
oligosaccharides (d.p. 2-20) by high-pressure
liquid chromatography. Carbohydr Res, 87, 165-
173.
70. Yun, J.W. (1996): Fructooligosaccharides—
Occurrence, preparation, and application.
Enzyme and Microbial Technology, 19, 107-117.