LWT - Food Science and Technology 65 (2016) 543e548

Contents lists available at ScienceDirect

LWT - Food Science and Technology

journal homepage: www.elsevier.com/locate/lwt

The impact of onion juice on milk fermentation by Lactobacillus

acidophilus
Sha Li a, b, Chengjie Ma a, b, *, Guangyu Gong a, Zhenmin Liu a, Chao Chang c, Zhiping Xu a, b
a

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
b

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 consump
bert, 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;

* Corresponding author. State Key Laboratory of Dairy Biotechnology, Technology

Center of Bright Dairy & Food Co., Ltd., 1518 Jiangchang Road (W), Shanghai 200436,
China.
E-mail address: chengjiema99@gmail.com (C. Ma).
http://dx.doi.org/10.1016/j.lwt.2015.08.042
0023-6438/ 2015 Published by Elsevier Ltd.

Talwalkar & Kailasapathy, 2003). Ng, Yeung, and Tong (2011)

found that the toxic oxygenic metabolites produced by yogurt
starter cultures were critical factors in impairing the survival of
L. acidophilus NCFM. Because of this, there are an increasing number of researchers attempting to produce fermented milk only with
L. acidophilus and to maintain or promote the viability of
L. acidophilus throughout the product shelf life by making the
environment more favorable to L. acidophilus. Hervert-Hern

andez,
~ i (2009) found that grape pomace
Pintado, Rotger, and Gon
phenolic extract induced a signicant biomass increase of
L. acidophilus growth. More recently, Oliveira, Perego, Oliveira, and
Converti (2011) found that inulin could improve the growth of
L. acidophilus in fermented skim milk.
Some researchers have found that a major part of phytochemicals are not absorbed in the small intestine, but that they can
interact with colonic microbiota, stimulating the growth of lactic
acid bacteria (LAB) and having antimicrobial activity against many
pathogens (Scalbert & Williamson, 2000). Najgebauer-Lejko, Sady,
Grega, and Walczycka (2011) reported that the addition of tea
extract containing polyphenols had a positive effect on yogurt
starter microora. Additionally, Michael, Phebus, and Schmidt

544

S. Li et al. / LWT - Food Science and Technology 65 (2016) 543e548

(2010) found that plant extracts possessing antioxidant capacity

extended the longevity of Lactobacillus delbrueckii subsp. bulgaricus
during storage.
Onions (Allium cepa L.) are one of most widely consumed vegetables in Europe (mainly in UK, Holland and Spain) and Asia
countries (mainly in China, India and Japan). Onion adequately
processed could be useful in the food industry to be added to foods
due to the increasing demand by consumers for substituting synthetic compounds by natural substances as functional food ingredients. Compounds of inherently natural origin would be widely
accepted by consumers in the market (Jang, Sanada, Ushio, Tanaka,
& Ohshima, 2002). There is an increasing interest in onions due to
their high level of phytochemicals. Among these, avonoids, fructans, and organosulfur compounds are considered important
contributing factors to antioxidant activity (Siddiq, Roidoung, Sogi,
& Dolan, 2013). For these reasons, the onion has been widely
researched for its antioxidant functions and antimicrobial properties against pathogenic microorganisms (Albishi, John, Al-Khalifa, &
Shahidi, 2013; Lee, Jung, & Kim, 2012). The onion may exert a
double positive effect on fermented milk production, on the one
hand stimulating LAB and inhibiting pathogenic bacteria, and on
the other hand improving nutritional values and health effects.
However, the stimulatory role of onions on LAB, and especially
L. acidophilus, has rarely been investigated. In addition, organosulfur compounds have a distinct avor and are identied as the
characteristic avor compounds in onions (Gyawali et al., 2006).
Adding onion juice to milk, therefore, may provide the benet of
adding avor to fermented milk.
In this study, onion juice was added to milk to investigate the
effects of onion juice on the growth of L. acidophilus NCFM. The
antioxidant capacity and avor compounds of fermented milk were
also evaluated.

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.

2. Materials and methods

2.6. Determination of volatile compounds
2.1. Preparation of onion juice
The fresh onion (commercially available) was cleaned and cut
into pieces, and then mixed with same amount of sterile distilled
water by weight. The initial onion juice was extracted from the
mixture using a Philips juicer (HR 1861, Philips China Investment
Co., Ltd., Zhuhai, China), and then centrifuged (10,000
g, 10 min).
The supernatant was ltered to obtain the nal onion juice used in
the experiment. This juice was refrigerated at 4  C until use.
2.2. Preparation of fermented milk
The fermented milk was prepared according to Lucey (2004)
with slight modications. Onion juice (0, 20, 60, and 100 g/kg)
was added into fresh cow's milk, and distilled water, respectively at
100, 80, 40, and 0 g/kg, was added into the onion juice to standardize the levels of protein and fat. The mixtures were stirred with
a stirrer RW20 (IKA, Staufen, Germany) at 600 rpm for 5 min at
65  C, and then homogenized under high pressure (18 MPa) at 65  C
and heat-treated at 95  C for 90 min immediately in order to kill all
the microorganisms, bacterial spores and fungal spores in milk and
onions due to low growth activity of L. acidophilus in milk. Cooled to
37  C, samples (each sample of two replicates) were inoculated
with L. acidophilus NCFM (Danisco, New Century, KS, USA) at
5
106 CFU/g. A set of samples were maintained at 37  C in an MIR253 incubator (Sanyo, Osaka, Japan). Another set of samples were
monitored automatically (recorded every 5 min) during milk
acidication for pH using a Cinac system (Alliance Instruments,
Mery-Sur-Oise, France), and the corresponding sample were
removed to terminate fermentation until a pH of 4.50 0.03 (the

The fermented milk containing 60 g/kg onion juice and the

control fermented milk were chosen for the detection of volatile
compounds.
Headspace solid-phase micro-extraction (SPME) was used to
extract volatile compounds from the fermented milk samples according to Li et al. (2014). The ber used for manual extraction was
DVB/CAR/PDMS (Supelco, Bellefonte, PA, USA).
Analysis of the volatile compounds was performed using an
Agilent 7890 (II) gas chromatograph (Agilent Technologies, Santa
Clara, CA, USA) coupled to an Agilent 5975 series mass selective
detector. The SPME ber was inserted into the injection port that
was held at 250  C, and the compounds were thermally desorbed
for 4 min under split (1:10) conditions (Welty, Marshall, Grn, &
Ellersieck, 2001). A DB-Wax column 30 m
0.25 mm
0.25 mm
(Agilent Technologies, Santa Clara, CA, USA) was used to separate
the volatile compounds. The temperature of the column was held at
45  C for 5 min, ramped up by 10  C/min to 80  C, and then further
heated to 240  C at the rate of 5  C/min. The carrier gas was helium
(1 mL/min). The mass spectrometer was run in the electron impact
mode at 70 eV. The mass scan range was 25e400 m/z (Condurso,
Verzera, Romeo, Ziino, & Conte, 2008).
2.7. Statistical analysis
All data were reported as the mean or mean standard deviation. Physicochemical experiments were carried out in triplicate. The
data was analyzed with ANOVA using Statistica 9.2 software (Stat
Soft, Inc., Tulsa, OK, USA). The comparison between means was
carried out using the Tukey's signicant difference test (p < 0.05).

S. Li et al. / LWT - Food Science and Technology 65 (2016) 543e548

3. Results and discussion

3.1. Effects of onion juice addition on acidication of L. acidophilus
The pH values of samples during fermentation are shown in
Fig. 1. With low levels (60 g/kg) of onion juice added, there was an
increase in acidication rates with increasing onion juice content.
However, when the content of onion juice reached 60 g/kg, the
maximum acidifying rate occurred. The results showed that addition of onion juice into fresh cow's milk exerted a stimulatory effect
on fermentation.
A possible explanation for the stimulatory effect was that
L. acidophilus possessed the ability to utilize compounds supplied
by onion juice during growth, such as phytochemicals, poly
ndez et al.
saccharides, vitamins, and/or minerals. Hervert-Herna
(2009) found that grape polyphenols induced a signicant
biomass increase of L. acidophilus during growth in liquid culture
media. However, not all plant extracts that were able to inhibit
pathogenic bacteria could stimulate fermentation by LAB. Yang
et al. (2012) found that ginger juice would inhibit the growth of
Streptococcus thermophilus and Lactobacillus bulgaricus and could
reduce the production of lactic acid.
The TA values of samples during cold storage are illustrated in
Fig. 2. The TA of all samples increased over the 14 days of storage.
The more onion juice that was added into the milk base, the
signicantly higher (p < 0.05) the TA of the sample was after
storage. However, there was not a signicant difference (p > 0.05)
in TA when the onion juice content was higher than 60 g/kg. These
results indicated that adding onion juice may promote the production of acid even at high acid concentrations and low
temperatures.
According to previous studies, L. acidophilus possesses several
mechanisms to counteract acid stress (Altermann et al., 2005;
Azcarate-Peril, Altermann, Hoover-Fitzula, Cano, & Klaenhammer,
2004). Kullen and Klaenhammer (1999) reported that the F1F0ATPase system maintaining the pHi was an important mechanism
for promoting survival in the presence of accumulating organic
acids.

545

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

3.2. Effect of onion juice addition on viable cell counts of

L. acidophilus
The viable cell counts of L. acidophilus in samples during storage

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.

546

S. Li et al. / LWT - Food Science and Technology 65 (2016) 543e548

Fig. 3. The viabilities of L. acidophilus in samples after 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.

Romero, Matallana Gonzalez, & Torija Isasa, 2008), which might

also account for the positive effects of its addition to milk. Babu,
Mital, and Garg (1992) reported that the addition of tomato juice
to skimmed milk stimulated the growth of L. acidophilus because of
sugars and minerals present in the juice. Sharma et al. (2013) reported that Mn-metabolite complexes could protect essential
enzymes from oxidative damage. Therefore, it is probable that the
whole constituents of onion juice such as fructans, polyphenols,
sulfur compounds, and minerals together played the stimulatory
role and resulted in this benecial effect for L. acidophilus viability
during fermentation and storage.
3.3. Antioxidant capacity of fermented milk
The AOCs of samples are presented in Fig. 4. It is noteworthy that
the AOC decreased signicantly (p < 0.05) after the milk was fermented by L. acidophilus NCFM (for example, AOC 0.783 before
fermentation vs. AOC 0.545 after fermentation in the control fermented milk; AOC 0.836 before fermentation vs. AOC 0.578 after
fermentation in the fermented milk containing 60 g/kg onion juice).
It is possible that some antioxidant compounds were consumed and
considerable oxygenic metabolites were produced simultaneously
as L. acidophilus NCFM grew in milk. The results conrmed that it
was very necessary to scavenge radical for L. acidophilus growth. The
AOC of fermented milk containing onion juice was slightly higher
than that of control fermented milk (p < 0.05), which suggested that
antioxidants in onions might played a certain role in growth and
maintenance of L. acidophilus NCFM viability.
Some previous studies have conrmed that the high level of
antioxidant activity in onions was attributable to their avonoid
constituents, namely quercetin, kaempferol, myricetin, and cate
ndez et al. (2009) inferred
chin (Siddiq et al., 2013). Hervert-Herna
that L. acidophilus may be able to use polyphenols possessing
antioxidant functions as substrates. Hence, adding onion juice into
a milk base would be an effective method to increase the AOC of
fermented milk and to maintain the viability of L. acidophilus.
3.4. Volatile avor compounds
The volatile compounds detected in the fermented milk samples

and control samples are given in Table 1. Twenty-three compounds,

consisting of 4 alcohols, 5 aldehydes, 5 carboxylic acids, 3 ketones,
and 6 sulfur compounds, were identied in the 4 samples.
As seen in Table 1, propanal, 2-methyl-2-butenal, and 6 sulfur
compounds that had been identied as main volatile compounds of
onions were detected in both the fermented sample (60 g/kg onion
juice) and its control sample (60 g/kg onion juice, not fermented by
L. acidophilus). Moreover, comparing the sulfur compound contents
in the two samples, most sulfur compounds had signicantly
decreased after fermentation (p < 0.05). The results conrmed that
L. acidophilus was able to utilize these compounds from onion juice
to supply energy to L. acidophilus during growth.
Acids produced in fermentation are the major components
causing the sour taste of fermented milk (Tamime & Robinson,
2007). Many carboxylic acids were detected in the fermented
milk samples. However, none of them was detected in the control
samples (not fermented by L. acidophilus). Previous studies have
shown that butanoic acid and hexanoic acid were produced by
lactose fermentation, and that hexanoic acid could also be formed
by lipolysis of free fatty acids (Ott, Fay, & Chaintreau, 1997). In
addition, comparing the contents of acids in the two fermented
milk samples, there were more carboxylic acids in the fermented
sample with 60 g/kg of onion juice added. The stimulatory effect of
onion juice on producing acid observed in this investigation by
might be attributed to avonoid, fructans and minerals, consistent
with the result of acidication of L. acidophilus.
Flavonoids are one of the abundant phenolic-type bioactive
compounds commonly found in vegetables and fruits, which been
categorized as avonoid, avanol, avanone, avone, anthocyani
ska, & Biesaga, 2007). The total
din, and isoavon (Wach, Pyrzyn
avonoid concentration was obtained at 89.6 mg/kg fresh onions by
summing the concentrations corresponding to all the identied

n, Rodrguez Rodrguez, & Daz
avonoids (Rodrguez Galdo
Romero, 2008). Fructans such as oligosaccharides and fructooligosaccharides are the other bioactive compounds in onion with the
total concentration of about 18 mg/kg of fresh weight (Rodrguez

n, Tasco

n Rodrguez, Rodrguez Rodrguez, & Daz Romero,
Galdo
2009), which with falvonoids show a variety of pharmacological
effects in stimulating the metabolism of microbial cells, including
L. acidophilus (China et al., 2012; Kaplan & Hutkins, 2000). Several

Fig. 4. The antioxidant capacity (AOC) of samples: , before fermentation; D day 1; G

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.

S. Li et al. / LWT - Food Science and Technology 65 (2016) 543e548

547

Table 1
Results of volatile compounds analysis of four samples, showing the averaged peak areas (in arbitrary unit
1000).
Compound

Averaged peak area
1000

Sample 1

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.

other interesting studies concerning the ability of L. acidophilus to

n-Huerta, Jua

rez-Flores,
growth in the presence of fructans. Rendo
Pinos-Rodrguez, Aguirre-Rivera, and Delgado-Portales (2011) reported that the lower pH values of the medium and the higher cell
counts were achieved in vitro growth of L. acidophilus with fructans
of Cichorium intybus, Helianthus tuberosus and Agave angustifolia
spp. tequilana. Pehrson, Mancilha, and Pereira (2014) reported that
L. acidophilus strain grew well in the medium containing fructans as
main carbohydrate sources, which a cellular viability signicantly
higher in this group than in the negative control.
Alcohol, aldehyde, and ketone are also important to the avor of
fermented milk. Comparing the alcohol and ketone contents in the
two fermented milk samples, there were more of these compounds
in the fermented sample with 60 g/kg of onion juice added, but the
aldehyde content was the opposite: more was found in the control
sample. Aldehyde is labile and has a high level of activity. When the
fermented milk was supplemented with onion juice having
considerable antioxidant substances, part of the aldehyde might
have been reduced to alcohol or other compounds. Overall, adding
onion juice into milk increased the volatile compounds in fermented milk and changed their avor compounds prole.
4. Conclusion
The onion juice signicantly stimulated the growth and acidication of L. acidophilus NCFM. The maximum acidifying rate was
observed for when 60 g/kg of onion juice was added to the fermented milk sample. Probably, the whole constituents of onion
juice together played the stimulatory roles and resulted in this
benecial effect for L. acidophilus viability. During cold storage, not
only did adding onion juice promote the production of acids by
L. acidophilus NCFM, but it also exerted a positive effect on the
maintenance of L. acidophilus NCFM viability. Hence, rather than
the accumulation of lactic acid, the accumulation of toxic oxygenic
metabolites may be the main reason for impaired survival of
L. acidophilus NCFM during storage. Onion juice was benecial to
shortening the fermentation time, enriching the avor components

and slightly enhancing the antioxidant capacity of fermented milk.

This study showed a potential application of adding onion juice to
the fermented milk production process.
Acknowledgments
This work was supported by National High Technology Research
and Development Program of China (2011AA100901), and the National Key Technologies Program of China (2013BAD18B01) during
the 12th Five-Year Plan Period.
References
Albishi, T., John, J. A., Al-Khalifa, A. S., & Shahidi, F. (2013). Antioxidative phenolic
constituents of skins of onion varieties and their activities. Journal of Functional
Foods, 5, 1191e1203.
Altermann, E., Russell, W. M., Azcarate-Peril, M. A., Barrangou, R., Buck, B. L.,
McAuliffe, O., et al. (2005). Complete genome sequence of the probiotic lactic
acid bacterium lactobacillus acidophilus NCFM. Proceedings of the National
Academy of Sciences USA, 102, 3906e3912.
Azcarate-Peril, M. A., Altermann, E., Hoover-Fitzula, R. L., Cano, R., &
Klaenhammer, T. R. (2004). Identication and inactivation of genetic loci
involved with lactobacillus acidophilus acid tolerance. Applied and Environmental
Microbiology, 70, 5315e5322.
Babu, V., Mital, B. K., & Garg, S. K. (1992). Effect of tomato juice addition on the
growth and activity of Lactobacillus acidophilus. International Journal of Food
Microbiology, 17, 67e70.
China, R., Mukherjee, S., Sen, S., Bose, S., Datta, S., Koley, H., et al. (2012). Antimicrobial activity of Sesbania grandiora ower polyphenol extracts on some
pathogenic bacteria and growth stimulatory effect on the probiotic organism
Lactobacillus acidophilus. Microbiological Research, 167, 500e506.
Condurso, C., Verzera, A., Romeo, V., Ziino, M., & Conte, F. (2008). Solid-phase
microextraction and gas chromatography mass spectrometry analysis of dairy
product volatiles for the determination of shelf-life. International Dairy Journal,
18, 819e825.
Donkor, O. N., Henriksson, A., Vasiljevic, T., & Shah, N. P. (2006). Effect of acidication on the activity of probiotics in yoghurt during cold storage. International
Dairy Journal, 16, 1181e1189.
Gilliland, S. E., & Speck, M. L. (1977). Instability of Lactobacillus acidophilus in yogurt.
Journal of Dairy Science, 60, 1394e1398.
Gyawali, R., Seo, H., Lee, H., Song, H., Kim, D., Byun, M., et al. (2006). Effect of
geirradiation on volatile compounds of dried welsh onion (Allium stulosum L.).
Radiation Physics and Chemistry, 75, 322e328.
~ i, I. (2009). Stimulatory role of
Hervert-Hern

andez, D., Pintado, C., Rotger, R., & Gon
grape pomace polyphenols on Lactobacillus acidophilus growth. International

548

S. Li et al. / LWT - Food Science and Technology 65 (2016) 543e548

Journal of Food Microbiology, 136, 119e122.

Jang, M. S., Sanada, A., Ushio, H., Tanaka, M., & Ohshima, T. (2002). Inhibitory effects
of 'enokitake' mushroom extracts on polyphenol oxidase and prevention of
apple browning. LWT-Food Science and Technology, 35, 697e702.
Kailasapathy, K., Harmstorf, L., & Philips, M. (2007). Survival of Lactobacillus acidophilus and Bidobacterium animalis ssp. lactis in stirred fruit yogurts. LWTFood Science and Technology, 41, 1317e1322.
Kaplan, H., & Hutkins, R. W. (2000). Fermentation of fructooligosaccharides by lactic
acid bacteria and bidobacteria. Applied and Environmental Microbiology, 66,
2682e2684.
Kullen, M., & Klaenhammer, T. R. (1999). Identication of the pH-inducible, protontranslocating F1F0-ATPase (atpBEFHAGDC) operon of Lactobacillus acidophilus
by differential display: gene structure, cloning and characterization. Molecular
Microbiology, 33, 1152e1161.
Lee, B., Jung, J., & Kim, H. (2012). Assessment of red onion on antioxidant activity in
rat. Food and Chemical Toxicology, 50, 3912e3919.
Li, S., Ma, C., Liu, Z., Gong, G., Xu, Z., Xu, A., et al. (2014). Flavour analysis of stirred
yoghurt with Cheddar cheese adding into milk. Food Science and Technology
Research, 20, 939e946.
Lucey, J. A. (2004). Cultured dairy products: an overview of their gelation and
texture properties. International Journal of Dairy Technology, 57, 77e84.
Michael, M., Phebus, R. K., & Schmidt, K. A. (2010). Impact of a plant extract on the
viability of Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus in nonfat yogurt. International Dairy Journal, 20, 665e672.
Najgebauer-Lejko, D., Sady, M., Grega, T., & Walczycka, M. (2011). The impact of tea
supplementation on microora, pH and antioxidant capacity of yoghurt. International Dairy Journal, 21, 568e574.
Ng, E. W., Yeung, M., & Tong, P. S. (2011). Effects of yogurt starter cultures on the
survival of Lactobacillus acidophilus. International Journal of Food Microbiology,
145, 169e175.
Oliveira, R. P. S., Perego, P., Oliveira, M. N., & Converti, A. (2011). Effect of inulin as a
prebiotic to improve growth and counts of a probiotic cocktail in fermented
skim milk. LWT- Food Science and Technology, 44, 520e523.
Ott, A., Fay, L. B., & Chaintreau, A. (1997). Determination and origin of the aroma
impact compounds of yogurt avor. Journal of Agricultural and Food Chemistry,
45, 850e858.
Pehrson, M. E. S. F., Mancilha, I. M., & Pereira, C. A. S. (2014). Mixture of short-chain
and long-chain fructans exert a positive effect on the growth of three potentially probiotic Lactobacillus strains. European International Journal of Science
and Technology, 3, 72e78.

bert, E. M., Mozzi, F., & de Valdez, G. F. (2010). Functional fermented
Pescuma, M., He
whey-based beverage using lactic acid bacteria. International Journal of Food
Microbiology, 141, 73e81.

n-Huerta, J. A., Jua

rez-Flores, B. I., Pinos-Rodrguez, J. M., Aguirre-Rivera, J. R.,
Rendo
& Delgado-Portales, R..,E. (2011). Effects of different kind of fructans on in vitro
growth of Lactobacillus acidophilus, Lactobacillus casei and Bidobacterium lactis.
African Journal of Microbiology Research, 5, 2706e2710.

n, B., Rodrguez Rodrguez, E. M., & Daz Romero, C. (2008). FlaRodrguez Galdo
vonoids in onion cultivars (Allium cepa L.). Journal of Food Science, 73,
C599eC605.

n, B., Tasco

n Rodrguez, C., Rodrguez Rodrguez, E. M., & Daz
Rodrguez Galdo
Romero, C. (2009). Fructans and major compounds in onion cultivars (Allium
cepa). Journal of Food Composition and Analysis, 22, 25e32.

n, E., Sanchez-Moreno, C., de Ancos, B., & Cano, M. P. (2008). Characterisation
Rolda
of onion (Allium cepa L.) by-products as food ingredients with antioxidant and
antibrowning properties. Food Chemistry, 108, 907e916.
Sanders, M. E., & Klaenhammer, T. R. (2001). Invited review: the scientic basis of
Lactobacillus acidophilus NCFM functionality as a probiotic. Journal of Dairy
Science, 84, 319e331.
Scalbert, A., & Williamson, G. (2000). Dietary intake and bioavailability of polyphenols. Journal of Nutrition, 130, 2073Se2085S.
Sharma, A., Gaidamakova, E. K., Matrosova, V. Y., Bennett, B., Daly, M. J., &
Hoffman, B. M. (2013). Responses of Mn2 speciation in Deinococcus radiodurans
and Escherichia coli to geradiation by advanced paramagnetic resonance
methods. Proceedings of National Academy of Sciences of the United States of
America, 110, 5945e5950.
Siddiq, M., Roidoung, S., Sogi, D. S., & Dolan, K. D. (2013). Total phenolics, antioxidant properties and quality of fresh-cut onions (Allium cepa L.) treated with
mild-heat. Food Chemistry, 136, 803e806.
Talwalkar, A., & Kailasapathy, K. (2003). Metabolic and biochemical responses of
probiotics bacteria in oxygen. Journal of Dairy Science, 86, 2537e2546.
Tamime, A. Y., & Robinson, R. K. (2007). Biochemistry of fermentation. In yoghurt
science and technology (3rd ed., pp. 535e577). Cambridge, UK: Woodhead
Publishing Ltd.
Verde Mendez, C. M., Rodriguez Rodriguez, E. M., Diaz Romero, C., Matallana
Gonzalez, M. C., & Torija Isasa, M. E. (2008). Comparison of the mineral and
trace element concentrations between 'gazpacho' and the vegetables used in its
elaboration. International Journal of Food Sciences and Nutrition, 59, 660e670.

ska, K., & Biesaga, M. (2007). Quercetin content in some food and
Wach, A., Pyrzyn
herbal samples. Food Chemistry, 100, 699e770.
Welty, W. M., Marshall, R. T., Grn, I. U., & Ellersieck, M. R. (2001). Effects of milk fat,
cocoa butter, or selected fat replacers on avour volatiles of chocolate ice
cream. Journal of Dairy Science, 84, 21e30.
Yang, G., Guan, J., Wang, J., Yin, H., Qiao, F., & Jia, F. (2012). Physicochemical and
sensory characterization of ginger-juice yogurt during fermentation. Food Science and Biotechnology, 21, 1541e1548.