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Impact of pasteurization and homogenization on the
digestion of human milk: an in vivo study in the
preterm infant
Samira Cassia de Oliveira, Olivia Ménard, Amandine Bellanger, Patrick
Pladys, Yann Le Gouar, Gwenaele Henry, Emelyne Dirson, Florence
Rousseau, Frédéric Carrière, Didier Dupont, et al.
To cite this version:
Samira Cassia de Oliveira, Olivia Ménard, Amandine Bellanger, Patrick Pladys, Yann Le Gouar, et
al.. Impact of pasteurization and homogenization on the digestion of human milk: an in vivo study
in the preterm infant. 15. Euro Fed Lipid Congress Oil, Fats and Lipids: New Technologies and
Applications for a Healthier Life, Aug 2017, Uppsala, Sweden. 2017. �hal-01583187�
Impact of pasteurization and homogenization
on the digestion of human milk: an in vivo
study in the preterm infant
SAMIRA DE OLIVEIRA
1
, OLIVIA MÉNARD
1
, AMANDINE BELLANGER
2,3
, PATRICK PLADYS
2,3
, YANN LE GOUAR
1
, GWENAËLE HENRY
1
,
EMELYNE DIRSON
2
, FLORENCE ROUSSEAU
1
, FRÉDERIC CARRIÈRE
4,
, DIDIER DUPONT
1
, AMÉLIE DEGLAIRE
1,
CLAIRE BOURLIEU
1
www6.rennes.inra.fr/stlo
1 STLO, Agrocampus Ouest, INRA, Rennes, France
2 CHU Rennes, Pediatrics department, France
3 University of Rennes 1, Faculty of Medicine, France
4 CNRS, Aix Marseille University, UMR7282 EIPL, France
* claire.bourlieu-lacanal@inra.fr
Materials & Methods
To investigate the impact
of the homogenization and
pasteurization of human milk
on its gastric digestion in
preterm infants
Objective
Context
Ensuring adequate growth of preterm newborns remains a challenge. When
breastfeeding not possible
pasteurized human milk from milk banks is
preferentially administered. Holder pasteurization (62.5°; 30 min) is applied for
sanitary reasons but may reduce fat absorption through the inactivation of milk
endogenous lipases. Conversely, homogenization of Holder-pasteurized human milk
(PHM) may improve fat absorption and weight gain.
©Thierry Pasquet
NCT02112331
Randomized controlled trial
Hospitalized tube-fed preterm infants
(
GA < 32 wks)
6-day experimental period; 2 independent groups
Experimental design
Multi-scale characterization of HM and digested samples
Results
Indirect homogenization by
ultrasonication
595 W, 3 periods of 5 min interrupted
by 30s of pause
Holder pasteurization
2 test meal/day, for each test meal, two gastric effluent collections:
before the meal and 35, 60 or 90 min after the ingestion start
Impact on
digestive kinetics?
(Thomaz et al., 1999; Rayol et al., 1993; Martinez et al., 1987)
HOMOGENIZED
HM
RAW HM
SDS-Page + densitometry;
Cation exchange
chromatography
Biochemical composition: kinetics of lipolysis and proteolysis
Structural disintegration
-1
0
1
2
3
4
5
6
7
8
9
10
0.01 0.1 1 10 100 1000 10000
%
vo
l.
Taille (µm)
Profil granulométrique M1 T
M1 T0
M1 Témoin 30 min
M1 Témoin 60 min
M1 Témoin 120 min
M1 Témoin 180 min
Confocal
microscopy
Laser light
scattering
Gas and thin layer
chromatography
Human Milk
analyzer
Instantaneous lipolysis level:
% of free fatty acids vs. total acyl chains present at a
given time
% of intact protein remaining at a given time:
Lactoferrin, α-lactalbumin, serum albumin, β-casein
(data not shown here)
Fluorescent probes
:
Proteins
(FastGreen®),
apolar
lipids
(Lipidtox®) and polar lipids (Rhodamine-PE®)
Particle size distribution
Six-fold increase in the specific
surface area after homogenization:
from
4.1 ± 1.2 m
2
/g
to
25.5 ± 3.8 m
2
/g
of fat
Homogenization increased gastric lipolysis
The initial structure of HM modulated the hydrolysis kinetics (lipolysis and proteolysis) and the emulsion disintegration during the gastric digestion in preterm infants
Overall, in vivo data gathered here are crucial for a better understanding of milk neonatal digestion and help supporting the nutritional management of preterm infants.
Past HM and P+Homog HM (n = 4 pools): same macronutrients
composition, same pre-lipolysis degree (4.4 ± 1.0%) but
different structure
Results were expressed as means ± SD
P < 0.001 (***); P < 0.01 (**); P < 0.05 (*);
P > 0.05 (NS).
0
4
8
12
16
20
HM
35
60
90
Lipolysis
degree (
%)
Time (min)
Meal: **
; Time: ***
Meal * Time: NS
Instantaneous lipolysis level
Past HM
P+Homog HM
(PHM: n=38 points;
P+Homog: n=36 points)
Homogenization affected the initial structure and the emulsion disintegration of HM
The lipid fraction kept its initial structure all over the
gastric digestion (
native globules
vs.
blend of droplets)
Particle size distribution in HM
Homogenization reduced the meal emptying rate
% of ingested meal remaining in the stomach
0
20
40
60
80
100
0
30
60
90
Time (min)
Meal: ***
Time: ***
Meal * Time: NS
0-30 min: HM ingestion
%
of
in
gested
meal
(PHM: n=36 points;
P+Homog: n=32 points)
T ½, half-emptying of
the ingested meal.
Hormonal feedback triggered by higher lipolysis level
Difference of colloidal behavior
Conclusion
GROUP A
HM from their own mother
collected < 24h
before feeding
1 pool aliquoted in 6 bottles
Raw HM
Pasteurized
HM
(PHM)
n=12, GA 30.0 ± 1.1 wk, age at first day 27 ± 12 d, Body
weight at first day 1.83 ± 0.41 kg
Pasteurization affected the emulsion disintegration of HM
GROUP A
0
4
8
12
16
20
HM
35
60
90
Lip
oly
sis
degr
ee
(%
)
Time (min)
Raw HM
Pasteurized HM
Meal: NS ; Time: ***
Meal*Time: NS
***
Instantaneous lipolysis level
But did not impact gastric lipolysis
GROUP B
T ½ = 30 min
T ½ = 38 min
Pre-lipolysis: n = 4 infants
Raw HM
= 2.2 ± 0.8%
Pasteurized HM
= 3.2 ± 0.6%
BSSL action
Heat-inactivation
Pasteurized
HM
(PHM)
Pasteurized +
Homogenized HM
HM from anonymous donor
GROUP B
The same
pool
from one donor was used for
the two types of milk
n=8, GA 29.5 ± 1.5 wk, age at first day 32 ± 21 d, Body
weight at first day 1.73 ± 0.48 kg
Titrimetry
Lipolytic activity in gastric aspirates:
Titration of butyric acid released from tributyrin at pH 4.5
and 37 °C (Gargouri et al., 1986)
0
2
4
6
8
0.01 0.1
1
10
100 1000
Volume
(%)
Size (μm)
Past HM
P+Homog
HM
(Time in min)
HM
35
60
90
10 µm
10 µm
10 µm
0
2
4
6
8
0.01 0.1
1
10
100 1000
V
olu
me
(%
)
Size (µm)
milks
Digestas
T60
Raw HM
Pasteurized
HM
10 µm
(Time in min)
HM
35
60
90
Particle size distribution
10 µm
10 µm
Pre-lipolysis occurred
prior to pasteurization
in all milks and groups
ranging between
2.2-4.0%. The global gastric
lipolytic
activity
detected in fasted state
averaged
17.5±2.9
U/mL/kg at pH 4.5.
Postprandial lipolytic
activity increased with
time and was higher
after administration of
Raw HM compared to
Past HM (n=5) whereas
it was not different
after administration of
Past HM vs. P+Homog
HM (n=4)
Lipolytic activity
