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Article history:
Received 15 September 2015
Received in revised form
15 March 2016
Accepted 20 March 2016
Available online 31 March 2016
Manufacture of ice cream using cold- and heat-adapted Lactobacillus acidophilus was studied. Temperatureetime combinations at 4 C for 18 h and at 45 C for 15 min were set as the adaptation conditions
for below and above the optimum growth temperature (37 C), respectively. Ice cream was produced by
two different methods: method 1, ice cream mix was fermented with cold- and heat-adapted
L. acidophilus prior to freezing; method 2, cold- and heat-adapted L. acidophilus was added to ice
cream mix but the mix was not fermented with L. acidophilus prior to freezing. The lowest reduction ratio
was found in the samples produced by using method 1 and cold-adapted L. acidophilus, adapted at 4 C
for 18 h. L. acidophilus survived at the required levels (>106 cfu g1), with or without an adaptation. The
adaptation conditions improved stability of L. acidophilus in the samples, but the magnitude of
improvement was small.
2016 Elsevier Ltd. All rights reserved.
1. Introduction
Consumer interest in different types of probiotic food products
has signicantly increased in recent years due to their nutritional
value and health-promoting properties (Casarotti & Penna, 2015).
Although many scientic studies have demonstrated some benets
to the consumption of probiotic cells, European Food Safety Association (EFSA) rejected nearly every health claim assert by the
probiotics industry due to insufcient proof of their health claims
(Kent & Doherty, 2014; Schmidt, 2013). However, manufacture of
probiotic products continues to expand, as do studies investigating
cell viability and functional properties of probiotic foods (Burgain,
Gaiani, Linder, & Scher, 2011). Among food products, ice cream is
known to be an advantageous vehicle to deliver probiotic bacteria
to human body since it has a neutral pH and high total solid level
providing protection for probiotic cells (Homayouni, Azizi, Ehsani,
Yarmand, & Razavi, 2008). However, cells must survive freezing
as well as frozen storage; temperature changes during freezing and
thawing may cause damage such as reduction or even complete
loss of metabolic activity (Akn, Akn, & Krmac, 2007; Haynes &
73
74
Fig. 1. Schematic illustration of the experimental setup for the adaptation tests.
2.5. Analyses for the ice cream mixes and ice creams
The pH values of the mixes and ice creams were measured using
a pH-meter (Thermo Scientic Orion 2-Star, Bremen, Germany).
Percentage of titratable acidity for the samples was measured by
the method of Bradley et al. (1992). The viscosity value of the mix
used in the ice cream production was measured using a Brookeld
viscometer (model DV-II; Brookeld Engineering Laboratories,
Inc., Stoughton, MA), with spindle 2, and speed at 1 rpm, following
the method described by El-Rahman, Madkor, Ibrahim, and Kilara
(1997) after 24 h of ripening at 4 C. Increase in volume (overrun)
was determined using a 100 mL cup and Eq. (1) described by
Voulasiki and Zerridis (1990):
% Overrun
The ice cream samples were analysed for dry matter content by
drying samples at 102 2 C for 3.5 h using an air oven, for fat
content using the Gerber method and for protein content by the
Kjeldahl method (Anonymous, 2003). Melting rate was estimated
according to El-Nagar, Clowes, Tudoric, Kuri, and Brennan (2002)
with slight modications. For the measurement of melting rate,
100 g of tempered samples were left to melt (at a constant temperature 15.5 0.3 C) on a 2.5 mm wire mesh screen above a
beaker. The weight of drip was measured over a 60 min period.
Firmness analysis was conducted using a texture analyser (TA-XT2,
Stable Microsystems, Godalming, Surrey, UK) tted with a 5 mm
diameter stainless steel probe, setup to record the force used to
75
76
Fig. 2. Survival of Lactobacillus acidophilus adapted under low temperature conditions, 4 C (a, d), 10 C (b, e) and 20 C (c, f) for mid-exponential phase (a, b, c) and stationary phase
(d, e, f) after 1, 15 and 30 days of exposure at 20 C. Adaptation times were:
, non;
, 1 h;
, 18 h;
, 24 h.
77
Fig. 3. Survival of Lactobacillus acidophilus adapted under high temperature conditions, 45 C (a, d), 50 C (b, e) and 55 C (c, f) for mid-exponential phase (a, b, c) and stationary
phase (d, e, f) after 1, 15 and 30 d exposure at 20 C. Adaptation times were:
, none;
, 15 min;
, 18 h;
, 24 h.
Table 1
Effect of the manufacturing method of ice cream and the adaptation treatment of Lactobacillus acidophilus on the physicochemical properties of ice cream mixes and ice
creams.a
Ice cream mix
pH
Ice cream
0.00a
0.00b
12.6.0 1.5a
8.1 1.1b
0.10a
0.10a
0.20a
11.2 4.0a
9.9 3.9b
9.9 4.2b
5.53 0.02b
6.43 0.04a
0.34 0.02a
0.13 0.00b
14.41 0.29b
16.42 0.25a
15.34 1.02a
15.38 0.99a
15.53 1.11a
a
Method 1, Ice cream mix was fermented with cold- and heat-adapted L. acidophilus prior to freezing; method 2, Cold- and heat-adapted L. acidophilus was added to the ice
cream mix prior to freezing and the mix was not fermented. Values are expressed mean standard deviation, different superscript letters after values indicate signicant
differences using Duncan's multiple range test (P < 0.05).
10.5
10
9.5
9
8.5
8
7.5
7
1
Milk
30
60
Ice cream, storage time (days)
90
C
0.12b
0.19a
0.17a
0.07c
0.20b
0.15b
0.19ba
0.19a
0.21a
a
Method 1, ice cream mix was fermented with cold- and heat-adapted
L. acidophilus prior to freezing; method 2, cold- and heat-adapted L. acidophilus
was added to the ice cream mix prior to freezing and the mix was not fermented.
Values are expressed mean standard deviation, different superscript letters after
values indicate signicant differences using Duncan's multiple range test (P < 0.05).
78
10
9
8
7
6
5
4
3
2
1
0
1
30
60
90
Storage (days)
Fig. 5. The percentage reduction for L. acidophilus counts in probiotic ice cream
samples according to ice cream mixes during storage. Ice creams were produced by: A
and :, non-adapted, methods 1 and 2, respectively; - and , heat-adapted (45 C for
15 min, mid-exponential phase), methods 1 and 2, respectively; and C, cold adapted
(4 C for 18 h, mid-exponential phase), methods 1 and 2, respectively.
stresses encountered during gastrointestinal model system application, such as acid and bile stresses, is of signicant importance
due to the fact that adaptation of the cells to one stress could result
in improved resistance not just to that stress, but also to different
stresses.
Acknowledgements
This cooperative project between the Akdeniz University and
the University of Hohenheim was supported by the Scientic
and Technological Research Council of Turkey (TUBITAK, Project
number: 112 O 890), the Alexander von Humboldt Foundation
(Project number: DEU/1116797) and the Scientic Research Projects
Coordination Unit of Akdeniz University (Project number:
2013.01.0102.006).
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