International Journal of Engineering Trends and Technology (IJETT) Volume 5 number 2 - Nov 2013

ISSN: 2231-5381 http://www.ijettjournal.org Page 86

Impact Of Human Milk Oligosaccharide (HMO) In
Neonates
S.Divya
#1
, B.SureshSubramonian
#2

#
Student, B.Tech (Food Technology)
1
,

Professor
2
College of Food and Dairy Technology,TANUVAS
Koduvalli, Redhills, Chennai-52, India.


AbstractHuman milk oligosaccharides (HMOs)
represent a highly heterogeneous class of carbohydrates
which are widely accepted to be beneficial for breast fed
infants because of their assumed anti-inflammatory, anti-
infective and immune stimulating properties. Human milk
oligosaccharides (HMOs) are known to be prebiotics
which is stimulating the growth of beneficial intestinal
bacteria, and actas receptor analogs by inhibiting the
binding of pathogens with cell surface glycans. In
addition, the development of a balanced intestinal micro
florasystem plays an important role in modulating the
postnatal immune system. And this article also describes
about the various local effects and systemic effects of
human milk oligosaccharide (HMO).The glycobiology of
the oligosaccharide and its impact study is reviewed.
Key words human milk oligosaccharide (HMO),
prebiotic, Anti-adhesive, glycobiology, probiotic, host
epithelial cell, neonates.
I. Introduction
uman Milk Oligosaccharide (HMO) is the premiere
prebiotics for the neonates. Human Breast milk is rich in
oligosaccharides and act as a source of commensal and
probiotic bacteria which seems to play an important role in
gut colonization and modulation of the infant gut [1].Human
milk oligosaccharide (HMO) plays an important role in
protecting neonates from pathogenic micro-organism
especially the enteric bacteria. This is due to their ability to
act as soluble receptors that inhibit the bacteria binding to
their host cell target ligands. HMO not only acts as substrates
for specific bacterial species, but also resembles their
adhesion molecules or ligands which may enable them to
directly affect gut maturation or influence inflammatory
processes.



A. Human milk oligosaccharide
In breast-fed babies, Human milk is often the sole
dietary source for the first few months of life. It contains all
the nutrients necessary for the infant to thrive, but also
ingredients that may provide health benefits beyond those of
traditional nutrients. Human milk oligosaccharides (HMO)
comprise part of these functional ingredients and nearly 1
Litre of mature human milk contains approximately 510 g
unbound oligosaccharides, and over one hundred and thirty
different HMO have been identified.
Oligosaccharide is the third most abundant
component of human milk. Human milk oligosaccharides
play a clinical role in protecting the neonates from infant
diarrhea as a result of Campylobacter, caliciviruses, and
diarrhoea of all causes in the breast-fed infants [2].

TABLE I

Micro nutrients of Human milk in g/l is compared with the
Bovine Milk:

Micro nutrients Human
milk(g/l)
Bovine
milk(g/l)
Protein 10 35
Fat 35 35
Lactose 65 45
Oligosaccharide 5-10* 0.05

The building blocks of Human milk oligosaccharides
are the 5 monosaccharides such as D-glucose (Glc), D-
galactose (Gal), N-acetylglucosamine (GlcNAc), L-Fucose
(Fuc), and sialic acid [(N-acetyl neuraminic acid (Neu5Ac)].
HMO act as a potential probiotic and immunogenic
compounds which is resistant to low pH and resistant to
pancreatic and brush border enzymes.
H
International Journal of Engineering Trends and Technology (IJETT) Volume 5 number 2 - Nov 2013
ISSN: 2231-5381 http://www.ijettjournal.org Page 87


Fig.1. Structure of HMO

.
The important functions of human milk oligosaccharides
are Prebiotic, Anti-adhesive, Anti-Microbial and Glycome
modifications. At present the modern methods have been
developed for the analysis of oligosaccharide. Mass
spectrometry and Nano flow liquid chromatography allow
rapid profiling for saccharide composition. There are nearly
200 different oligosaccharides in human milk while in other
mammals number is less than 50 and often less than 20. Only
trace amounts of these oligosaccharides are present in mature
bovine milk and, as a consequence, it is low in bovine milk
based infant formula. Recent advances in glycobiology and
nutrition which includes the use of stable isotopes, frontal-
affinity chromatography, glycan microarrays, MS, and
automated solid-phase carbohydrate synthesis, will help
verify hypotheses and unravel the mysteries behind HMO.

II. Effects of HMO
A. Local effects
Once the human milk is ingested into the human
system, HMO can withstand the low pH in the gut and resist
degradation through enzymes from the pancreas and brush
border membrane [3], [4]. Therefore, intact HMO rinses the
infant's esophagus, stomach, and small intestine before they
serve as nutrients for colon bacteria. As in [5] it was reported
about the prebiotic effects of HMO showing that a mixture of
HMO which promotes the growth of Bifidobacterium
bifidum.
Other oligosaccharides, structurally different from
those in human milk, have also been added to the infant
formula to mimic potential prebiotic effects as in [6], [7].


Adhesion to the host's epithelial surface is essential
for the virulence of most pathogenic microorganisms, e.g.
Campylobacter jejuni, Escherichia coli, Vibrio cholera, and
Shigella and Salmonella strains and C. jejuni, is one of the
most predominant causes of diarrhoea worldwide. The
adhesion-related virulence factors are often lectins,
carbohydrate-binding proteins, which dock to
oligosaccharides on the epithelial cell surface. These
carbohydrate-binding determinants are also often part of
HMO, suggesting that HMO serve as soluble ligand analogs,
block pathogen adhesion, and protect breast-fed infants
against infections and diarrhea [8]. Fucosylated HMO will
inhibit the growth of Campylobacter species binding to
human intestinal mucosa [9]. Antimicrobial effects of HMO
were also described for calicivirus diarrhea [10] and
infections with heat-stable enterotoxin of E. coli as in [11].
HMO may change the glycome of intestinal
epithelial cells. 3sialyllactose (3SL) is one of the
predominant sialylated HMO which reduces the expression of
various glycosyltransferases, which diminishes the content of
cell surface sialic acid, fucose, and galactose. Such
glycocalyx modifications may alter the ability of certain
pathogens to adhere. 3SL exposure of Caco-2 cells reduces
the adhesion of enteropathogenic E. coli (EPEC) by 50%
[12].
HMO rinses the laryngopharyngeal region and may also
reduce pathogen adhesion at the entry to the upper respiratory
tract. Human milk inhibits adhesion of Streptococcus
pneumoniae and Haemophilusinfluenzae to human
pharyngeal mucosa. These pathogens attach to sialylated
glycans on the host's epithelium and are responsible for most
of the otitis media cases and respiratory tract infections in
newborn infants [13]. Sialylated glycans are also part of
HMO and may partially account for the beneficial effects of
human milk as in [14].

B. Systemic effects
HMO is partially absorbed intact in the infant's
intestine and appears in the urine of breast-fed, but not
formula-fed infants. HMO serves as anti-adhesive receptor
analogs for urinary pathogens. And the appearance of HMO
in the urine indirectly proves their presence in the systemic
circulation. They may alter protein-carbohydrate interactions
also on a systemic level. Selectins, for example, are involved
in cell-cell interactions in the immune system [15]. P-selectin
is also involved in the formation of platelet-neutrophil
complexes (PNC), a subpopulation of highly activated
neutrophils primed for adhesion, phagocytosis, and enhanced
production of reactive oxygen species (ROS) [16].HMO
inhibit leukocyte adhesion at sites of inflammation and reduce
formation of highly active PNCs, they may contribute to the
International Journal of Engineering Trends and Technology (IJETT) Volume 5 number 2 - Nov 2013
ISSN: 2231-5381 http://www.ijettjournal.org Page 88

protection of breast-fed infants against NEC and other
inflammatory diseases as in [17], [18].

1) Control of amebiasis: Breast-fed infants are at
lower risk to develop such devastating disorders such
asamebiasis or necrotizing enterocolitis (NEC) and
worldwide, approximately 50 million people are infected with
E. histolytica, resulting in nearly 100,000 deaths annually.
E. histolytica resides in the colon, where it used as a specific
Gal/GalNAclectin to attach to and destroy the hosts epithelial
cells. Here, the specific HMO but also
Galactooligosaccharides (GOS) prevent E.histolytica
attachment and cytotoxicity. These results suggest that HMO
contribute to the lower incidence of amebiasis in breast-fed
infants compared to formula-fed infants.
III. Bioactive HMO
HMO is a heterogenic group of at least 130 different
compounds. Which of these oligosaccharides interact with a
given lectin is currently unknown and as per the above recent
methods we can profile the HMO glycome and describe inter-
individual differences as well as intra-individual changes
during the time of lactation. Once individual bioactive
HMO is identified, they can be chemically synthesized and
used to supplement infant formula for clinical trials. The
recent development of automated solid-phase oligosaccharide
synthesis has been a milestone for efficient and fast chemical
HMO synthesis [19].
Some of the natural sources of HMO like oligosaccharide
were Goat's milk is a rich source of HMO-like
oligosaccharides and may be suitable for supplementing
formula. These oligosaccharides separate as by-products in
goat cheese production and can be isolated by membrane
technology on a large scale [20].
IV. In-Vitro studies on effect of HMO
The effect of the human milk oligosaccharide fraction
(HMOS) on infant fecal microbiota cultured in vitro was
investigated to determine the prebiotic activity of HMO. It
supplemented cultures which is developed significantly at
lower pH and after fermentation, HMO supplemented
cultures contained higher lactic acid concentrations. HMO
supplemented microbiota contained significantly more
Bifidobacteria and Lactobacillus sp. By this activity the use of
HMO supplemented microbiota will significantly decrease the
number of E. coli, Clostridium perfringens, and Clostridium
difficile. When HMO is supplemented in-vitro, then the
growth of Bifidobacteria and Lactobacilli were stimulated,
and it inhibits the growth of clostridial pathogens. HMOS also
inhibit binding by C. perfringensto intestinal epithelial cells.
These changes in microbiota community composition are
highly reminiscent of differences in microbiota of breastfed
infants relative to microbiota of prematurely weaned infants
[21].
A.Carbohydrates as future anti-adhesion drugs for
infectious diseases
Sharon Nathan, (2006) Adhesion of pathogenic
organisms to host tissues is the prerequisite for the initiation
of the majority of infectious diseases. In many systems, it is
mediated by lectins present on the surface of the infectious
organism that bind to complementary carbohydrates on the
surface of the host tissues. Lectins-deficient mutants often
lack the ability to initiate infection. The bacterial lectins are
typically in the form of elongated submicroscopic multi-
subunit protein appendages, known as fimbriae (or pili).
Soluble carbohydrates recognized by the bacterial surface
lectins block the adhesion of the bacteria to animal cells in
vitro. And type 1 fimbriated E.Coli has been inhibited by
aromatic -mannosides and this is 1000 times more active due
to the presence of hydrophobic region next to the
monosaccharide-binding site of the fimbriae. The suitable
sugars will also inhibit the binding property with the cells of
carbohydrate-specific toxins, among them those of
Shigelladysenteriae Type 1, and of the homologous
Verotoxins of E. coli, specific for galabiose. The above data
provide clear proof for the feasibility of anti-adhesion therapy
of infectious diseases using CHO.
V. Benefits of breast-feeding compared to formula
feeding
Less diarrhea
Fewer and less severe episodes of infection
Higher intelligence
Less necrotizing enterocolitis
Less atopy
Possible protection from sudden infant death
syndrome
Long term health effects

VI. Conclusion
There is increasing evidence that HMOs are of particular
importance for the infant health. Functions which are
discussed include anti-adhesive and anti-inflammatory
effects, role as probiotics and it has an influence on brain
development or preventive effects with regard to certain
diseases and this oligosaccharide influence the composition of
intestinal micro biota and allow the growth of bifidobacterium
and lactobacillus organism. It is obvious to conclude that
HMO has wider functional importance in the biological
system and to maintain the infants in healthy state of being.
VII. Reference
[1] Coppa, Giovanni V; Bruni, Stefano; Morelli, Lorenzo; Soldi,
Sara; Gabrielli, Orazio. The First Prebiotics in Humans: Human
Milk Oligosaccharides, J ournal of Clinical
Gastroenterology;38(2),S80-S83, 2004.
[2] Ardythe.L. Morrow,Guillermo M. Ruiz-Palacios, Mekibib Altaye,
Xi J iang, M. Lourdes Guerrero,J areen K. Meinzen-
Derr,Tibor Farkas, Prasoon Chaturvedi,Larry K. Pickering, David
S. Newburg, Human milk oligosaccharides are associated with
protection against diarrhea in breast-fed infants, J ournal of
pediatrics, 145( 3 ), pp. 297-303, 2004
International Journal of Engineering Trends and Technology (IJETT) Volume 5 number 2 - Nov 2013
ISSN: 2231-5381 http://www.ijettjournal.org Page 89

[3] Gnoth MJ , Kunz C, Kinne-Saffran E, Rudloff S, Human milk
oligosaccharides are minimally digested in vitro. J
Nutr. ;130(12):3014-20; 2000 Dec
[4] Engfer MB, Stahl B, Finke B, Sawatzki G, Daniel H. Human milk
oligosaccharides are resistant to enzymatic hydrolysis in the
upper gastrointestinal tract. Am J ClinNutr; 71:158996, 2000.

[5] Gyorgy P, Norris RF, Rose CS. Bifidus factor I. A variant of
Lactobacillus bifidus requiring a special growth factor. Arch
BiochemBiophys ; 48:193201, 1954

[6] Euler AR, Mitchell DK, Kline R, Pickering LK. Prebiotic effect of
fructo-oligosaccharide supplemented term infant formula at two
concentrations compared with unsupplemented formula and
human milk. J. Pedi atr. Gastroenterol. Nutr. 40:15764,
2005.

[7] Knol J , Scholtens P, Kafka C, Steenbakkers J , Gro S, HelmK,
Klarczyk M, Schopfer H, Bockler HM, Wells J . Colon microflora
in infants fed formula with galacto- and fructo-oligosaccharides:
more like breast-fed infants. J .Pediatr.Gastroenterol.Nutr.; 40:36
42, 2005

[8] Newburg DS, Ruiz-Palacios GM, Morrow AL. Human milk
glycans protect infants against enteric pathogens. Annu Rev
Nutr; 25:3758, 2005

[9] Ruiz-Palacios GM, Cervantes LE, Ramos P, Chavez-Munguia B,
Newburg DS. Campylobacter jejuni binds intestinal H(O) antigen
(Fuc alpha 1, 2Gal beta 1, 4GlcNAc), and
fucosyloligosaccharides of human milk inhibit its binding and
infection. J .Biol.Chem. 278:1411220, 2003

[10] Morrow AL, Ruiz-Palacios GM, Altaye M, J iang X, Guerrero
ML, Meinzen-Derr J K, Farkas T, Chaturvedi P, Pickering LK,
Newburg DS. Human milk oligosaccharides are associated with
protection against diarrhea in breast-fed infants. J Pediatr.
145:297303; 2004

[11] Newburg DS, Ruiz-Palacios GM, Altaye M, Chaturvedi P,
Guerrero ML, Meinzen-Derr J K, Farkas T, Chaturvedi P,
Pickering LK, Newburg DS. Human milk alphal,2-linked
fucosylated oligosaccharides decrease risk of diarrhea due to
stable toxin of E. coli in breastfed infants. Adv Exp Med Biol.
554:45761, 2004.
[12] Angeloni S, Ridet J L, Kusy N, Gao H, Crevoisier F, Guinchard S,
Kochhar S, Sigrist H, Sprenger N. Glycoprofiling with micro-
arrays of glycoconjugates and lectins. Glycobiology. 15:3141,
2005.
[13] Andersson B, Porras O, Hanson LA, Lagergard T, Svanborg-Eden
C. Inhibition of attachment of Streptococcus pneumoniae and
Haemophilusinfluenzae by human milk and receptor
oligosaccharides. J .Infect. Dis.153:2327; 1986.

[14] Dewey KG, Heinig MJ , Nommsen-Rivers LA. Differences in
morbidity between breast-fed and formula-fed infants. J Pediatr.
126:696702, 1995.


[15] Ley K. The role of selectins in inflammation and disease. Trends.
Mol. Med.9:2638; 2003.

[16] Peters MJ , Dixon G, Kotowicz KT, Hatch DJ , Heyderman RS,
Klein NJ . Circulating platelet-neutrophil complexes represent a
subpopulation of activated neutrophils primed for adhesion,
phagocytosis and intracellular killing. Br. J .Haematol; 106:391
9; 1999.


[17] Lucas A, Cole TJ . Breast milk and neonatal
necrotisingenterocolitis. Lancet.336:151923; 1990.

[18] Bode L, Kunz C, Muhly-Reinholz M, Mayer K, Seeger W,
Rudloff S. Inhibition of monocyte, lymphocyte, and neutrophil
adhesion to endothelial cells by human milk oligosaccharides.
ThrombHaemost. 92:140210; 2004.
[19] Plante OJ , Palmacci ER, Seeberger PH. Automated solid-phase
synthesis of oligosaccharides. Science. 291:15237; 2001.

[20] Martinez-Ferez A, Rudloff S, Guadix A, Henkel CA, Pohlentz G,
Boza J J , GuadixEm, Kunz, C. Goat's milk as a natural source of
lactose-derived oligosaccharides: isolation by membrane
technology. Int. Dairy. J . 16:174178; 2006.


[21] zhuo-tengyu and david s. newburg,Abstract on Human milk
oligosaccharides alter human faecalmicrobiota during in vitro
fermentation ,The First International Conference on the
Glycobiology of Human Milk Oligosaccharides May 16 & 17,
2011 Tivoli Hotel, Copenhagen, Denmark.

[22] Sharon Nathan,carbohydrate as future.e anti-adhestion drugs for
infectious disease; Biochimi ca et Bi ophysica Acta
vol. 1760, n
o
4, pp. 527-537 , 2006.