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State Key Laboratory of Dairy Biotechnology, Technology Center of Bright Dairy & Food Co., Ltd., 1518 Jiangchang Road (W), Shanghai 200436, China
Wuhan Bright Dairy Co., Ltd., 1 Zhangbai Road, Wuhan 430040, China
c
College of Food Science and Engineering, Wuhan Polytechnic University, 68 Xuefu Road (S), Changqing Garden, Wuhan, 430023, China
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a r t i c l e i n f o
a b s t r a c t
Article history:
Received 4 March 2015
Received in revised form
12 August 2015
Accepted 14 August 2015
Available online 17 August 2015
The aim of this study was to investigate the inuence of onion juice on the fermentation of milk by
Lactobacillus acidophilus. The onion juice signicantly stimulated the growth and acidication of the
bacterial strain L. acidophilus NCFM. The viable cell counts in onion-supplemented fermented milk were
higher than that of the control sample and were maintained during cold storage. We considered the
whole constituents of onion juice such as polyphenols, sulfur compounds, fructans, and minerals
together to play the stimulatory roles and result in this benecial effect for L. acidophilus viability during
fermentation and storage. After the milk was fermented by L. acidophilus NCFM, the antioxidant capacity
decreased greatly in the control and onion-supplemented fermented milk. The results suggested that it
was very necessary and crucial to scavenge radical for L. acidophilus growth. The onion juice was slightly
benecial to enhancing the antioxidant capacity of fermented milk. The higher cell numbers, the less
fermentation time, the better antioxidant capacity, and more abundant avor components were achieved
by onion-supplemented fermentation by L. acidophilus, demonstrating the potential for industrial
applications.
2015 Published by Elsevier Ltd.
Keywords:
Lactobacillus acidophilus
Onion juice
Viable cell count
Antioxidant capacity
Flavor compound
1. Introduction
Lactobacillus acidophilus NCFM is a common commercial bacterial strain used in the manufacturing of dairy products
(Kailasapathy, Harmstorf, & Philips, 2007). Research has shown that
it is capable of protecting the host by improving lactose digestion,
preventing or lessening the impact of diarrhea, improving blood
lipid chemistry, stimulating an immune response, and potentially
killing cancer cells as they develop (Sanders & Klaenhammer,
2001). To realize the health benets of L. acidophilus, an adequate
number of viable cells must be delivered at the time of consumpbert, Mozzi, & de Valdez, 2010). However, the
tion (Pescuma, He
viable cells of probiotics signicantly decrease during cold storage
because of acid accumulation, interaction with starter cultures,
levels of dissolved oxygen and hydrogen peroxide, and storage
conditions (Donkor, Henriksson, Vasiljevic, & Shah, 2006;
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pH end point) was reached. After samples were stirred at 500 rpm
for 5 min, they were stored at 4 C.
2.3. Determination of titratable acidity
The titratable acidity (TA) of fermented milk samples was
determined according to Yang et al. (2012) on day 1 and after 14
days of cold storage.
2.4. Enumeration of L. acidophilus
A standard plate count was used to enumerate viable
L. acidophilus cells according to Ng et al. (2011) with slight modications. Fermented milk samples were serially diluted using
sterile saline to achieve 20 to 200 colonies on MRS (de Man, Rogosa
and Sharpe) agar plates. L. acidophilus was cultured under anaerobic conditions (Bugbox Anaerobic System, Ruskinn, Bridgend,
United Kingdom) with a mixture of 95% N2 and 5% CO2 at 37 C for
72 h.
2.5. Determination of antioxidant capacity (AOC)
For the determination of the AOC of non-fermented milk or
fermented milk, the diphenyl picrylhydrazyl (DPPH) method according to Najgebauer-Lejko et al. (2011) was used in this study. It is
a simple, sensitive, and widely used method to measure the radical
scavenging activity of all the antioxidant substances (including
protein, glutathione, lipoic, etc.) present in the examined sample.
The amount of sample required to decrease the initial DPPH concentration by 50% (EC50) was calculated. Antioxidant capacity was
given as AOC, the reciprocal of EC50. The AOC was calculated relative to the AOC of Trolox, an analog of vitamin E, and expressed as
mmol/TEg of sample.
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are shown in Fig. 3. All onion juice supplemented samples contained signicantly higher levels of L. acidophilus than the control
sample (p < 0.05). Moreover, the viable cell counts of L. acidophilus
increased as the content of onion juice increased. In addition, there
was a signicant decrease in viable cell counts of L. acidophilus
during storage when the content of onion juice was less than 20 g/
kg (p < 0.05), while the fermented milk containing 60 g/kg or
higher of onion juice had no signicant decrease in viable cell
counts of L. acidophilus during cold storage (p > 0.05). Hence, the
onion juice had positive effects on the growth and maintenance of
L. acidophilus viability.
Several studies had conrmed the decrease in viability of probiotics attributable to acid accumulation, interaction with starter
cultures, levels of dissolved oxygen and hydrogen peroxide, and
storage conditions (Donkor et al., 2006; Talwalkar & Kailasapathy,
2003). Adding onion juice not only promoted the production of
acid but also increased the viable cell counts, which indicated that
acid accumulation was not an important factor contributing to the
loss of L. acidophilus NCFM cell viability during storage. In general,
L. acidophilus does not possess a sufcient scavenging mechanism,
so the intracellular accumulation of toxic oxygenic metabolites
such as superoxide anion and hydroxyl radical can lead to its death
(Gilliland & Speck, 1977; Talwalkar & Kailasapathy, 2003). Ng et al.
(2011) also reported that L. acidophilus does not possess a catalase,
an enzyme hydrolyzing H2O2 into water and oxygen, so that
hydrogen peroxide might be an important factor resulting in the
loss of cell viability of L. acidophilus NCFM.
Onion is rich in antioxidant phenolic constituents such as
quercetin 3,40 -diglucoside, quercetin, and kaempferol that are
considered important contributing factors to antioxidant activity
(Albishi et al., 2013). These compounds might scavenge hydrogen
peroxide and protect L. acidophilus during the fermentation and
storage. Hervert-Hern
andez et al. (2009) found polyphenols possessing antioxidant functions had a stimulatory effect on
L. acidophilus, and a possible explanation for this would be that
L. acidophilus was able to use these compounds as substrates. On
the other hand, onions are a source of fructooligosaccharides, sulfur
n, Sanchez-Moreno, de
compounds, vitamins, and minerals (Rolda
Ancos, & Cano, 2008; Verde Mendez, Rodriguez Rodriguez, Diaz
Fig. 1. Changes in the pH of samples added of onion juice (0 g/kg, 20 g/kg, 60 g/kg, and
100 g/kg) during fermentation.
Fig. 2. The titratable acidity of samples during storage: , day 1; D day 7; Q day 14;
bars with different letters denote signicant differences (p < 0.05) among the samples
with different contents of onion juice added. Error bars with different letters denote
signicant differences (p < 0.05) among the samples with different contents of onion
juice added and among the different storage periods.
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Table 1
Results of volatile compounds analysis of four samples, showing the averaged peak areas (in arbitrary unit 1000).
Compound
Hexanol
Heptanol
Octanol
Furanmethanol
Propanal
Hexanal
Octanal
Benzaldehyde
2-methyl-2-butenal
Propanoic acid
Butanoic acid
Hexanoic acid
Octanoic acid
Decanoic acid
Heptanone
Nonanone
Undecanone
Dimethyl disulde
Methyl propyl disulde
Methyl propenyl disulde
Dimethyl trisulde
Dipropyl trisulde
2, 4-Dimethyl thiophene
62.6
29.6
113.9
ND
ND
219.5
136.2
240.0
ND
ND
ND
ND
ND
ND
3262.5
3464.4
847.0
ND
ND
ND
ND
ND
ND
Sample 2
d
1.2
0.8d
4.7b
3.5c
1.7c
5.2c
30.1c
28.9a
7.5a
109.4
311.8
119.2
166.2
ND
421.5
264.2
659.3
ND
ND
724.4
2821.5
829.9
92.4
3060.1
1334.9
212.8
ND
ND
ND
ND
ND
ND
Sample 3
b
3.5
5.6b
2.8b
3.3a
7.6a
3.8a
6.9a
7.8b
24.4b
8.9b
1.3b
33.5d
11.8d
4.3d
73.9
42.1
93.2
ND
353.1
68.1
32.4
86.1
6999.9
ND
ND
ND
ND
ND
6160.1
2163.2
624.9
3266.1
1803.4
123.4
2256.2
255.0
2239.7
Sample 4
c
1.5
1.7c
1.7c
5.8a
0.6d
0.4d
1.1d
32.1a
37.6a
21.6b
6.7b
41.2a
15.6a
1.9b
22.5a
1.3a
22.5a
132.8
697.8
129.8
171.3
98.7
335.6
189.1
310.7
5656.6
107.5
790.6
3247.4
1341.2
190.5
4341.0
1475.5
368.1
1725.7
1581.4
237.1
1832.9
123.4
2158.8
4.8a
7.2a
1.5a
4.2a
1.8b
4.4b
1.4b
1.5b
28.7b
1.8
8.9a
28.9a
10.2a
2.1a
31.5b
19.9c
5.7c
31.5b
14.3b
2.1a
11.5b
0.9b
18.4b
All values are mean of three replications SD. Different superscript letters in the same row indicate signicant differences at P < 0.05. ND: not detected. Sample 1: 0 g/kg onion
juice & not fermented; Sample 2: 0 g/kg onion juice & fermented; Sample 3: 60 g/kg onion juice & not fermented; Sample 4: 60 g/kg onion juice and fermented.
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