Efficacy of slightly acidic electrolyzed water on the microbial safety and shelf
life of shelled eggs
Y. T. Zang, Sh. Bing, Y. J. Li, D. Q. Shu,1 A. M. Huang, H. X. Wu, L. T. Lan, and H. D. Wu
Jiangxi Agricultural University, Jiangxi 330045, China
ABSTRACT This study evaluated the efficacy of
slightly acidic electrolyzed water (SAEW) for the in-
activation of Salmonella Enteritidis and Escherichia
coli on shelled eggs as well as studied the shelf life
and internal quality attributes (i.e., weight loss, Haugh
unit, yolk index, albumen pH, and yolk pH) of eggs
during storage at 25◦ C. The decontamination test egg
samples (freshly laid) were inoculated and immersed
for 1, 2, 3, and 4 min in each treatment (i.e., SAEW,
acidic electrolyzed water, NaClO solution, and ster-
ile deionized water) at available chlorine concentra-
tions (ACCs) of 10, 18 and 26 mg/L. The storage
test eggs (freshly laid) were immersed for 3 min in 4
treatments (i.e., SAEW, acidic electrolyzed water, Na-
ClO solution, and no treatment) and stored for 30 D
at 25◦ C. The effects of the SAEW on microbiological
qualities and freshness parameters were investigated.
Decontamination of eggs with SAEW showed an equiv-
alent or higher bactericidal effect compared to other
Key words: shelled eggs, slightly acidic electrolyzed water, shelf life, microbial safety
INTRODUCTION
Eggs can be contaminated with many types of mi-
croorganisms during production and thus present a risk
for the transmission of foodborne disease to consumers
(De Reu et al., 2006; Cao et al., 2009; Chousalkar
et al., 2010). Salmonella and other bacteria can rapidly
penetrate the shell and contaminate internal contents
of the egg (De Reu et al., 2008). De Reu et al. (2006)
reported that Staphylococcus warneri, Acinetobacter
baumannii, Alcaligenes sp., Serratia marcescens,
Carnobacterium sp., Pseudomonas sp., and Salmonella
Enteritidis were able to penetrate the shell of an
egg most frequently after approximately 5 D. This
penetration may result in the deterioration of egg
quality during storage and cause a major economic
loss to the poultry industry (Berrang et al., 1998; Cox
et al., 1998; Holck et al., 2018). Numerous outbreaks

C 2019 Poultry Science Association Inc.
Received March 23, 2019.
Accepted June 25, 2019.
1
Corresponding author: 13607047771@163.com
5932
treatments. A complete inactivation of S. Enteritidis
and E. coli on the surface of shelled egg samples resulted
from treatment with SAEW at an ACC of 26 mg/L for
3 and 4 min, respectively. Almost all of the egg qual-
ity parameters investigated in the present study were
significantly (P < 0.05) affected by the storage time.
As storage time increased, the yolk index and Haugh
unit value decreased, and weight loss, albumen pH, and
yolk pH increased. However, SAEW treatment mini-
mized weight loss (5.52%) and preserved the albumen
and yolk quality better than no treatment at 25◦ C. Rel-
ative to acidic electrolyzed water and NaClO solution,
the advantages of SAEW are reduced corrosion of egg
surfaces and potentially less water and CO2 escaping
from eggshell pores. The results highlight the promising
use of SAEW to enhance the microbial safety and to
extend the shelf life of shelled eggs. Future combined
methods with SAEW and other treatments are also
needed.
2019 Poultry Science 98:5932–5939
http://dx.doi.org/10.3382/ps/pez373
of diseases in humans have been caused by Salmonella
and other bacteria from the consumption of contami-
nated eggs or their products (De Reu et al., 2008). In
2015, approximately 95,000 food-poisoning outbreak
cases of Salmonella were registered in the European
Union, and Salmonella in eggs accounted for 10% of all
strong-evidence outbreaks, which were associated with
the highest number of reported food outbreaks (De
Reu et al., 2008). Therefore, decontaminating eggshells
is required to improve the microbiological safety or to
extend the storage time of the eggs used for human
consumption.
Eggs can be contaminated with bacteria, and the
egg content is an ideal growth medium for pathogenic
bacteria, such as Salmonella, Escherichia or Enter-
obacter, which are hazardous to humans (Chousalkar
et al., 2010). Decontamination of the egg surface
would be a critical step in reducing foodborne illnesses
(Turtoi and Borda, 2014). Currently, most egg proces-
sors utilize chemical sanitization systems to decontam-
inate the surfaces of eggshells prior to packaging (Cao
et al., 2009). Some of these decontamination procedures
 SLIGHTLY ACIDIC ELECTROLYZED WATER ON EGGS
include treatments with chlorine and iodine (Berrang
et al., 2000), boiling water (Himathongkham et al.,
1999), and hydrogen peroxide (Cox et al., 2000).
However, some chemical solutions have not been com-
pletely acceptable because of chemical residue, limited
effectiveness, and adverse environmental impacts
(Cao et al., 2009). Slightly acidic electrolyzed water
(SAEW) with a pH value of 5.0 to 6.5 contains a high
concentration of hypochlorous acid (HOCl), and its
application is widely accepted as an environmentally
friendly sanitization method (Huang et al., 2008; Koide
et al., 2009; Zang et al., 2015).
In recent years, the use of SAEW as an egg surface
decontamination method has been met with increas-
ing interest (Cao et al., 2009; Ni et al., 2014). Some
studies have demonstrated that SAEW could be used
as a disinfectant in egg processing (Cao et al., 2009;
Ni, et al., 2014). Cao et al. (2009) reported that a re-
duction of 6.5 log10 CFU/g of S. Enteritidis on shelled
eggs was obtained by SAEW at 15 mg/L available chlo-
rine concentration (ACC) for 3 min. Ni et al. (2014)
also reported that the bactericidal activity of SAEW
on shelled eggs toward Escherichia coli O157: H7, S.
Enteritidis, total aerobic bacteria, molds, and yeasts
was significantly higher than that of chlorine dioxide
and NaClO solution at an ACC of 80 or 100 mg/L.
These findings indicate that SAEW may be an alterna-
tive disinfectant to reduce the population of pathogens
on shelled eggs. However, some studies reported that
water and many chemical solutions can be detrimental
to the quality of the egg by washing away the cuticle
surrounding the egg, and cuticle removal may affect the
quality and the shelf life of the product (Berrang et al.,
1998; Bialka et al., 2004).
Bialka et al. (2004) evaluated the efficacy of acidic
electrolyzed water (AEW) for the microbial safety and
quality of eggs and reported that AEW did not sig-
nificantly affect albumen height or eggshell strength.
However, it significantly (P < 0.05) affected the cu-
ticle, which may favor transshell contamination with
bacteria and moisture loss and may shorten the shelf
life of the egg (Messens et al., 2005). Therefore, it was
necessary to further investigate if SAEW could be used
to sanitize shelled eggs without affecting the shelf life
of the eggs. These elements are important for practical
implementation of SAEW technology in the industry.
However, little information is available on the efficacy
of SAEW to improve shelf life and protect eggs from
deterioration during storage.
The overall objectives of this study were (1) to com-
pare the efficacy of SAEW and other disinfectants (i.e.,
AEW and NaClO) for inactivation of S. Enteritidis and
E. coli on artificially inoculated shelled eggs with differ-
ent ACCs (i.e., 10, 18, and 26 mg/L) at different time
points (e.g., 1, 2, 3, and 4 min) and (2) to evaluate
the efficacy of SAEW treatment on the internal qual-
ity attributes (i.e., weight loss, Haugh unit (HU), yolk
5933
index, albumen pH, and yolk pH) of eggs during storage
at 25◦ C.
MATERIALS AND METHODS
Preparation of Bacterial Cultures
The strains of S. Enteritidis (CVCC 2184) and E.
coli (ATCC 25,922) were obtained from the China Vet-
erinary Culture Collection (CVCC, Beijing, China).
Each bacterium was hydrated according to the manu-
facturer’s directions and cultured in tryptic soy broth
(TSB; CVCC, Beijing, China) at 37◦ C for 24 h. Follow-
ing incubation, 10-mL culture was poured into a sterile
centrifuge tube and centrifuged at 4,000 × g and 4◦ C
for 10 min with a centrifuge (5418R;Eppendorf, Ham-
burg, Germany). The supernatant was decanted, and
the pellet was resuspended in 10 mL of 0.1% buffered
sterile peptone water (BPW; Beijing Land Bridge
Technology Company Ltd., Beijing, China), washed 3
times and resuspended in 10 mL of the same solu-
tion to obtain a final cell concentration of approxi-
mately 8 log10 CFU/mL. The bacterial population in
each culture was confirmed by plating 0.1 mL portions
of appropriately diluted culture on tryptic soy agar
(TSA; Beijing Land Bridge Technology Company Ltd.,
Beijing, China) plates and then incubating the plates
at 37◦ C for 24 h. The prepared cultures were then used
in subsequent experiments.
Preparation of Treatment Solutions
SAEW was produced using a nonmembrane genera-
tor (Ruiande Biosafety Technology Co., Ltd., Beijing,
China) to electrolyze a NaCl solution (1 g/L) contain-
ing HCl (100 μg/L). The SAEW generated was di-
luted in sterile deionized water to obtain different ACCs
(Table 1). AEW was produced by electrolysis of a 0.1%
NaCl solution in a chamber with a separating mem-
brane between the anode and cathode using a gen-
erator (Zhouji Ziyuan Huanbao Technology Company
Ltd., Beijing, China). AEW with a pH of 2.50 ± 0.41
and oxidation-reduction potential (ORP) of 1161.3 ±
3.2 mV and an ACC of 30 mg/L was collected from
the anode side of the AEW generator and then di-
luted in sterile deionized water to obtain the different
final ACCs (Table 1). In addition, the NaClO solution
was obtained from a company (Jinan Xiaosi Technol-
ogy Company Ltd., Jinan, China) and then diluted in
sterile deionized water to obtain the different final con-
centrations of NaClO solution (10, 18, and 26 mg/L)
(Table 1).
The pH, ORP, and ACC of treatment solutions
were measured immediately before each experiment.
The pH and ORP values were measured with a dual-
scale pH/ORP meter (CON60, Trans-Wiggens, Singa-
pore). The ACC was determined by a digital chlorine
5934 ZANG ET AL.

Table 1. Physicochemical properties of sodium hypochlorite solution (NaClO), acidic elec-

trolyzed water (AEW), and slightly acidic electrolyzed water (SAEW) for inactivation of
artificially inoculated shell eggs.

Available chlorine
Solutions concentration (mg/L) pH ORP1 (mV)

Control (deionized water) 0 6.16 ± 0.02a 397.8 ± 4.0a

NaClO solution 10 10.12 ± 0.04b 432.6 ± 2.0a
18 10.24 ± 0.03b 464.3 ± 5.0a
26 10.38 ± 0.04b 478.2 ± 3.0a
Acidic electrolyzed water 10 2.81 ± 0.03c 1097.7 ± 9.0c
18 2.66 ± 0.01c 1132.4 ± 7.0c
26 2.54 ± 0.02c 1156.3 ± 2.0c
Slightly acidic electrolyzed water 10 6.53 ± 0.01d 647.5 ± 3.0b
18 6.42 ± 0.01d 653.2 ± 5.0b
26 6.37 ± 0.02d 675.9 ± 7.0b
Values reported as the means of triplicate measurements ± standard deviation.
Values in each column followed by different lower-case letters in superscripts (a–d) are significantly
different (P < 0.05).
1
Oxidation reduction potential.

test system (RC-2Z, Kasahara Chemical Instruments
Co., Saitama, Japan). The detection range was 0 to
320 mg/L.
Design of Experiments
Preparation of Shelled Eggs Freshly laid, special
class unfertilized egg samples (white shell, from 70-wk-
old Hy-line W-36 hens) weighing 55 to 60 g were ob-
tained directly from the egg company (Qingshan Com-
pany Ltd., Nanchang, China) and transported to the
laboratory under temperature-controlled conditions in
a sterile plastic container.
The decontamination test eggs were first equilibrated
to room temperature before testing then sequentially
washed with tap water and a commercial chlorine-
based sanitizer (Beijing Zhenhe Medical Technology
Co. Ltd., Beijing, China) at an ACC of 30 mg/L for
1 min, washed with sterile deionized water to com-
pletely remove the sanitizer, and then air-dried under
a biosafety hood (DH-920, Beijing East Union Hall In-
strument Manufacturing Co., Ltd., Beijing, China). For
inoculation, eggs were individually soaked into the in-
oculum, prepared by placing 0.1 mL approximately 8
log10 CFU/mL S. Enteritidis or E. coli suspension into
200 mL sterile 0.1% peptone water for 10 min, and ster-
ilely air-dried under the biosafety hood for 60 min at
a room temperature of 25◦ C to allow bacterial attach-
ment. The final concentrations of S. Enteritidis and E.
coli inoculated on the egg surface were approximately
6.45 ± 0.24 and 6.27 ± 0.19 log10 CFU/g, respectively,
on average.
A total of 80 eggs for the storage test were ran-
domly divided into 4 groups: control (no treatment),
SAEW (30 mg/L), AEW (30 mg/L), and NaClO solu-
tion (30 mg/L). Each treatment’s immersion time was
3 min. After treatment, eggs were placed in plastic egg
trays, marked carefully before storage, and stored under
ambient conditions at 25 ± 2◦ C and a relative humidity
of 78 ± 4% for 30 D. Each sample was analyzed on days
0, 6, 12, 18, 24, and 30 at 25 ± 2◦ C.
Ten eggs for each treatment were prepared and sam-
pled at least twice.
Bacteriological Analysis of Shelled Eggs Deter-
mination of S. Enteritidis and E. coli counts on the
eggshell surface was carried out by following a previ-
ously reported method (Cao et al., 2009). Inoculated
shelled eggs were individually placed in a sterile plastic
bag containing 500 mL SAEW, AEW, or NaClO solu-
tion at an ACC of 10, 18, and 26 mg/L, respectively, or
sterile deionized water (control) for 1, 2, 3, and 4 min.
The bags were then shaken vigorously by hand at a
room temperature of 25 ± 2◦ C. After treatment, the
egg sample was placed into a sterile plastic bag, which
contained 50 mL sterile neutralizing buffer solution, and
shaken vigorously for 1 min. The viable bacterial popu-
lation in the washed treatment solutions and neutraliz-
ing buffer solution was serially diluted with sterile 0.1%
BPW. A volume of 0.1 mL of each S. Enteritidis and E.
coli dilution in triplicate on TSA plates and Violet Red
Bile with Glucose Agar (Qingdao Hope Bio-Technology
Co. Ltd., Qingdao, China) was plated and incubated at
37◦ C for 24 h. The shell was also weighed to deter-
mine the colony-forming units per gram of eggshell +
membrane (CFU/g) by following a previously reported
method (Bialka et al., 2004).
Weight Loss The weight was established by indi-
vidually weighting each marked egg using an analytical
balance (YJ-GS-600 g, Shanghai Yajin Electronic Tech-
nology Co., Ltd., Shanghai, China) with a sensitivity of
0.01 g. Weight loss (%) of the eggs during storage was
calculated by subtracting the final weight of the egg
from the initial weight and then dividing by the ini-
tial weight and multiplying by 100 (Caner and Yuceer,
2015).
Yolk Index and Haugh Unit The yolk index is a
measure of egg freshness and is based on the ratio of
yolk height to yolk width (Caner and Yuceer, 2015).
Eggs were cracked onto a flat glass surface with a
 SLIGHTLY ACIDIC ELECTROLYZED WATER ON EGGS 5935
Table 2. Inactivation of S. Enteritidis on the surface of shell eggs by slightly acidic electrolyzed water, sodium hypochlorite solution
(NaClO), and acidic electrolyzed water (AEW).

Surviving population of S. Enteritidis on eggs (log10 CFU/g)

Available chlorine
Treatment concentration (mg/L) 1 min 2 min 3 min 4 min

Control 0 5.58 ± 0.26a 4.99 ± 0.23a 4.38 ± 0.19a 3.87 ± 0.11a

SAEW2 10 4.47 ± 0.21c 3.35 ± 0.11c 2.16 ± 0.17c 1.08 ± 0.14c
18 3.91 ± 0.13c 2.5 ± 0.15c 1.02 ± 0.21c ND1
26 3.01 ± 0.17c 1.56 ± 0.13c ND1 ND1
NaClO solution 10 4.61 ± 0.06b 3.63 ± 0.18b 2.41 ± 0.25b 1.27 ± 0.17b
18 4.13 ± 0.18b 2.85 ± 0.14b 1.40 ± 0.22b ND1
26 3.51 ± 0.14b 1.79 ± 0.08b ND1 ND1
AEW3 10 4.52 ± 0.16b,c 3.55 ± 0.21b,c 2.34 ± 0.19b,c 1.19 ± 0.09b,c
18 4.05 ± 0.16b,c 2.78 ± 0.22b,c 1.21 ± 0.14b,c ND1
26 3.14 ± 0.12b,c 1.63 ± 0.21b,c ND1 ND1
The data are expressed as the means ± standard deviations.
Within the same column of different solutions at same available concentration, values with different lowercase letters in superscripts (a–c) within
a column were different (P < 0.05), and b,c means no difference with b and c .
1
Means not detected.
2
Means slightly acidic electrolyzed water.
3
Means acidic electrolyzed water.

spatula to measure various internal parameters. The
height and diameter of the yolk were measured using
a tripod micrometer (NFN381, Nanjing Mingao Instru-
ments and Equipment Co., Ltd., Nanjing, China) and a
digimatic caliper (500–150-30, Mitutoyo Co., Kawasaki-
shi, Japan), respectively.
The HU is related to albumen quality and based on
a function of the height of the inner thick albumen and
egg weight (Morsy et al., 2015). A fresh, good quality
egg would have an HU index of approximately 80, and
an older egg has a lower HU (Yuceer et al., 2016). The
HU was determined from the egg weight and albumen
height of a broken egg spread on a horizontal plate using
the formula (Haugh, 1937):
 
Haugh unit = 100 × log h − 1.7w0. 37 + 7.51
(1)
where h is the albumen height (mm) and w is the weight
of the egg tested (g).
The parameter, h, was recorded by averaging 3 mea-
surements carried out at different points of the thick
albumen at a distance of 10 mm from the yolk using the
tripod micrometer as described above. All the parame-
ters were measured with 3 replicates and were expressed
as means.
pH Measurement After the yolk and albumen were
separated, each egg was placed and homogenized in a
beaker. Then, pH values of the albumen and yolk were
measured using a pH meter (SP-2500, Suntex Instru-
ments Company Ltd., Jiangsu, China). Measurements
were performed with 3 replicates and average values
were reported.
Statistical Analysis
All experiments had 3 replications for each treatment
and measurement. Mean values of all parameters were
calculated from the independent triplicate trials. Statis-
tical analysis was performed using Origin (Version 9.0,
OriginLab Cor., Hampton). Differences between vari-
ables were assessed by the Tukey test. Results with
P < 0.05 were considered statistically significant.
RESULTS AND DISCUSSION
Inactivation of S. Enteritidis and E. coli on
the Surface of Shelled Eggs by SAEW and
Other Solutions
Tables 2 and 3 illustrate the survival characteristics
of S. Enteritidis and E. coli inoculated on the surface
of shelled eggs, treated by NaClO solution, AEW, and
SAEW with respective ACCs of 10, 18, and 26 mg/L for
1, 2, 3, and 4 min. The initial population of S. Enteri-
tidis and E. coli inoculated on the surface of shelled egg
samples was 6.45 ± 0.24 and 6.27 ± 0.19 log10 CFU/g,
respectively. The bactericidal efficiency of SAEW in-
creased with increasing ACCs and increasing treatment
times at 25◦ C. A complete inactivation of S. Enteritidis
and E. coli on the surface of shelled egg samples was
resulted in using NaClO solution, AEW, or SAEW at
ACC of 26 mg/L for 3 min. Compared to deionized wa-
ter, all treatment solutions reduced the population of S.
Enteritidis and E. coli (P < 0.05). The bactericidal ac-
tivity of SAEW was higher than that of NaClO solution
at the same ACC (P < 0.05). However, no differences
were observed between AEW and SAEW solutions at
the same ACC (P > 0.05). Our findings demonstrated
that SAEW has an equivalent or higher efficiency in
reducing S. Enteritidis and E. coli present on shelled
eggs compared to NaClO solution and has bactericidal
activities similar to AEW at the same ACC.
The same result was also obtained by Ni et al. (2014),
who reported that the bactericidal activity of SAEW
on shelled eggs for E.coli O157: H7 and S. Enteritidis
was significantly higher than that of NaClO solution
at ACCs of 80 and 100 mg/L (P < 0.05), and no
significant sanitization difference was shown between
5936 ZANG ET AL.

Table 3. Inactivation of E. coli on the surface of shell eggs by slightly acidic electrolyzed water (SAEW) sodium hypochlorite solution
(NaClO), and acidic electrolyzed water (AEW).

Surviving population of E. coli on eggs (log10 CFU/g)

Available chlorine
Treatment concentration (mg/L) 1 min 2 min 3 min 4 min

Control 0 5.35 ± 0.23a 4.79 ± 0.16a 4.24 ±0.13a 3.69 ± 0.21a

SAEW2 10 3.45 ± 0.12c 2.91 ± 0.14c 1.66 ± 0.05c 0.43 ± 0.02c
18 2.31 ± 0.12c 1.57 ± 0.11c 0.52 ± 0.01c ND1
26 1.21 ± 0.05c 0.27 ± 0.03c ND1 ND1
NaClO solution 10 3.58 ± 0.07b 2.13 ± 0.10b 1.69 ± 0.07b 0.57 ± 0.06b
18 2.55 ± 0.08b 1.64 ± 0.12b 0.69 ± 0.02b ND1
26 1.32 ± 0.02b 0.43 ± 0.08b ND1 ND1
AEW3 10 3.49 ± 0.11b,c 2.46 ± 0.11b,c 1.64 ± 0.07b,c 0.47 ± 0.04b,c
18 2.47 ± 0.13b,c 1.61 ± 0.12b,c 0.53 ± 0.04b,c ND1
26 1.27 ± 0.11b,c 0.36 ± 0.01b,c ND1 ND1
The data are expressed as the means ± standard deviations.
Within the same column of different solutions at same available concentration, values with different lowercase letters in superscripts (a–c) within
a column were different (P < 0.05), and bc means no difference with b and c .
1
Means not detected.
2
Means slightly acidic electrolyzed water.
3
Means acidic electrolyzed water.

SAEW and AEW at the same ACC. Cao et al. (2009)

also compared the efficiency of SAEW and other dis-
infectants (i.e., AEW and NaClO solution) to inacti-
vate S. Enteritidis on the surface of shelled eggs and
demonstrated that SAEW with a near-neutral pH value
exhibits equivalent or higher bactericidal activity for
shelled eggs compared to AEW and NaClO solution.
Yoshifumi (2003) reported that the effective form of
chlorine compounds in SAEW was mostly hypochlor-
ous acid (HOCl) because of its near-neutral pH value.
This finding means that SAEW has a higher HOCl con-
centration than AEW and NaClO solution at the same
ACC. Guentzel et al. (2008) reported that the antimi-
crobial activity of chlorine-related solutions depends on
the amount of HOCl present in the water, and it may
be the reason that SAEW exhibits bactericidal activity
equivalent to or higher than AEW and NaClO solution. Figure 1. Effect of disinfectants on the weight loss of eggs. SAEW:
Our findings indicated that SAEW has high bacte- slightly acidic electrolyzed water, AEW: acidic electrolyzed water. Val-
ues shown are means ± standard deviation.
ricidal activity against S. Enteritidis and E. coli for
shelled eggs, and it may be an alternative disinfectant
for shelled eggs. Moreover, previous work has demon- trans-shell contamination with bacteria and moisture
strated that the application of SAEW in food products loss from the egg during storage. To further investigate
could not only reduce the initial microbial load but also if SAEW could be utilized to sanitize shelled eggs with-
extend the shelf life of the food products (Rahman et out affecting the shelf life of the eggs, a more detailed
al., 2013; Mansur and Oh, 2015; Sheng et al., 2018). storage study was of interest.
Duan et al. (2017) evaluated the effects of various dis- Weight Loss Weight loss is an important parame-
infectant sprays on initial microbial load and conducted ter that was used to monitor the changes in the qual-
a shelf-life study of chilled chicken carcasses, and they ity of fresh shelled eggs during storage (Samli et al.,
found that SAEW spray was the most effective treat- 2005). The results of the relative weight loss from the
ment with reductions of 0.47 to 0.83 log10 CFU/cm2 and effects of storage time and disinfectants are shown in
0.49 to 0.96 log10 MPN/cm2 in total viable counts and Figure 1. The weight loss of eggs increased as the stor-
total coliforms, respectively. They found that chilled age time increased for all groups. Similar results were
chicken carcasses treated with SAEW had a microbial reported by Samli et al. (2005), Jin et al. (2013), Morsy
shelf life extension of 2 D compared to the controls. et al. (2015), and Yimenu et al. (2017). They all re-
Sheng et al. (2018) also reported that SAEW treat- ported that weights of the eggs decreased significantly
ment is an effective and promising method to prolong during storage. Moreover, different weight loss findings
the shelf life of beef by approximately 8 D at 4◦ C. in these studies may be due to different storage condi-
However, Bialka et al. (2004) found that EW signifi- tions, temperature, egg size, hen age, and shell porosity
cantly affects the egg cuticle and that effect may favor (Yimenu et al., 2017).
 SLIGHTLY ACIDIC ELECTROLYZED WATER ON EGGS
Figure 2. Effect of disinfectants on the yolk index of eggs. SAEW:
slightly acidic electrolyzed water, AEW: acidic electrolyzed water. Val-
ues shown are means ± standard deviation.
No differences between the control group, Na-
ClO solution-, and AEW-treated eggs were observed
(P > 0.05). However, there were differences among the
SAEW treatment group and other groups (P < 0.05).
The average weight loss for SAEW-treated eggs stored
at 25◦ C after 30 D was 5.52%, whereas weight losses
were 6.51, 6.42, and 6.47% for the control group and the
NaClO solution- and AEW-treated eggs, respectively.
Normally, weight loss of eggs during storage is mainly
caused by evaporation of water and loss of carbon diox-
ide from the albumen through the porous shells (Samli
et al., 2005). In our study, the 3 min immersion of the
egg in AEW and NaClO solution may have caused dete-
rioration of the cuticle and pores, resulting in increased
weight loss than SAEW-treated eggs. Relative to other
disinfectant treatment groups, the SAEW-treated eggs
may have reduced deterioration of the cuticle and po-
tentially slowed weight loss of the egg. Although we did
not conduct cuticle experiments in the present study,
numerous studies have documented that SAEW, with
a near-neutral pH value, has the extra advantage of
reduced corrosion of surfaces compared to AEW and
NaClO solutions (Cao et al., 2009; Zang et al., 2017).
Yolk Index Dramatic deteriorations were observed
in the yolk index due to the storage time as shown in
Figure 2. After 30 D of storage, the yolk index values
of control eggs were 0.26, while SAEW-, AEW-, and
NaClO solution-treated eggs had values of 0.33, 0.31,
and 0.30, respectively. The yolk index value, which is
used as an indicator of freshness, declines as a result
of a weakening of the vitelline membranes, reduction
of the total solid, and liquefaction of the yolk caused
mainly by osmotic diffusion of water from the albu-
men during storage (Morsy et al., 2015). Each disin-
fectant treatment of eggs had a lower yolk index than
untreated eggs after 30 D of storage at 25◦ C (P < 0.05).
Furthermore, the yolk index of eggs exposed to SAEW
treatment was higher than AEW and NaClO solution
after 18 D (P < 0.05), and no significant difference was
5937
Figure 3. Effect of disinfectants on the Haugh unit of eggs. SAEW:
slightly acidic electrolyzed water, AEW: acidic electrolyzed water. Val-
ues shown are means ± standard deviation.
observed between AEW and NaClO solution treatment
groups. According to the results, SAEW treatment ef-
fectively reduced the mass transfer rate (i.e., water and
CO2 loss) from the albumen through the eggshell dur-
ing 30 D of storage. Consequently, this process inhibits
albumen liquefaction and water uptake by the yolk and
diminishes a reduction in yolk quality. The eggs im-
mersed in SAEW at an ACC of 30 mg/L for 3 min
were able to preserve their yolk quality longer than the
control eggs at 25◦ C.
Haugh Unit Changes in the HU for the various
disinfectant-treated and the control (untreated) eggs
are shown in Figure 3. The HU decreased in all groups
during storage (P < 0.05). This result is in agreement
with those of previous publications (Jin et al., 2013;
Caner and Yuceer, 2015; Yimenu et al., 2017). The
reduction in the HU was induced by the decrease in
the thick albumen height, which becomes thinner and
loses CO2 during storage, and it allows the electrostatic
lysozyme-ovomucin complex to rupture, which helps the
pH of eggs increase (Morsy et al., 2015).
All disinfectant-treated eggs exhibited a higher HU
than the control eggs during 30 D of storage at 25◦ C
(P < 0.05). After 30 D of storage, the HU of the
control eggs was 40.8, while SAEW-, AEW-, and Na-
ClO solution-treated eggs had values of 55.3, 50.7, and
49.1, respectively. This finding indicates that sterilizing
treatments minimized egg white thinning during stor-
age. A similar result was shown by Yuceer et al. (2015),
who indicated that ozone can slow down the decrease of
the HU (P < 0.05), and the function was increased with
the increasing concentration of the ozone. Morsy et al.
(2015) also demonstrated that treatment of unwashed
eggs with antimicrobial pullulan-based coating exhib-
ited a slightly higher HU than pullulan-based coated
eggs. Moreover, the HU index of eggs exposed to SAEW
treatment was higher than AEW and NaClO solution
after 12 D (P < 0.05), and no difference was observed
between AEW and NaClO solution treatment groups
5938 ZANG ET AL.
Figure 4. Effect of disinfectants on the albumen pH of eggs. SAEW:
slightly acidic electrolyzed water, AEW: acidic electrolyzed water. Val-
ues shown are means ± standard deviation.
Figure 5. Effect of disinfectants on the yolk pH of eggs. SAEW:
slightly acidic electrolyzed water, AEW: acidic electrolyzed water. Val-
ues shown are means ± standard deviation.
(P > 0.05). This result demonstrated that SAEW treat-
ment can preserve albumen quality during storage at
25◦ C.
pH Measurement As seen from Figures 4 and 5,
significant increases in albumen and yolk pH were ob-
served with increasing time. Albumen pH can also be
used as an indicator of egg freshness except for the yolk
index and HU (Yuceer et al., 2016). Normally, freshly
laid eggs have an albumen pH that lies between 7.6 and
8.5 (Yuceer et al., 2016), but during storage, carbon
dioxide (CO2 ) from the breakdown of carbonic acid in
albumen escapes via eggshell pores, resulting in changes
in the bicarbonate buffer system and increases in the
albumen pH up to 9.6 (Caner and Yuceer, 2015).
In this study, the albumen pH of untreated eggs
at 0 D was approximately 8.21, but during storage
of the eggs, the pH gradually increased to 9.55. For
disinfectant-treated eggs, yolk pH values reached 9.02,
9.19, and 9.17 for SAEW, NaClO solution, and AEW,
respectively, after 30 D storage time. Furthermore, the
albumen pH values were higher for untreated eggs than
for the disinfectant-treated eggs during storage (P <
0.05). According to these results, eggshell sterilizing, es-
pecially with SAEW, was helpful to maintain albumen
quality by controlling the albumen pH. There were dif-
ferences among the SAEW treatment group and other
disinfectant groups (P < 0.05). The major reason is
that SAEW may lead to lower deterioration of the cu-
ticle compared to NaClO solution and AEW and poten-
tially slows down the CO2 escape from eggshell pores.
In addition, no differences were observed between the
AEW and NaClO solution treatment groups (P > 0.05).
As shown in Figure 5, overall increases in the pH of
the yolk were observed at the end of all the storage ex-
periments. After 30 D of storage, the yolk pH of the con-
trol eggs reached 6.56, while SAEW-, AEW-, and Na-
ClO solution-treated eggs had values of 6.43, 6.45, and
6.46, respectively. There were differences among the
SAEW treatment group and other disinfectant groups
(P < 0.05). These findings showed that SAEW treat-
ment was effective in maintaining the pH of fresh eggs
during storage.
In conclusion, this study demonstrated that SAEW
exhibits equivalent or higher bactericidal activity for
shelled eggs compared to AEW and NaClO solutions
and is not only effective in reducing or eliminating S.
Enteritidis and E. coli on shelled eggs but also could
maintain fresh egg quality during storage. Relative to
AEW and NaClO solutions, the advantages of SAEW
are reduced corrosion of egg surfaces and potentially
less water and CO2 escape from eggshell pores. SAEW
can be used to preserve eggs at 25◦ C by inhibiting bac-
terial penetration into the albumen and yolk. Further-
more, the potential of SAEW to preserve the albumen
quality and maintain the yolk index suggests its effi-
cacy in preventing egg spoilage. However, although it
is an improvement over other disinfectants in terms of
eggshell cuticle deterioration, it still causes a slightly
adverse impact. Therefore, new combined methods such
as SAEW and coating, SAEW and UV light, or SAEW
and refrigeration storage for extending the shelf life of
eggs also need to be investigated to avoid this negative
aspect and to improve the effectiveness of egg preser-
vation.
ACKNOWLEDGMENTS
The authors gratefully acknowledge the financial sup-
port from the National Natural Science Foundation of
China (31860665).
CONFLICT OF INTEREST STATEMENT
The authors declare that there are no conflicts of
interest.
REFERENCES
Berrang, M. E., N. A. Cox, J. F. Frank, R. J. Burh, and J. S. Bailey.
2000. Hatching egg sanitization for prevention or reduction of
 SLIGHTLY ACIDIC ELECTROLYZED WATER ON EGGS
human enteropathogens: a review. J. Appl. Poult. Res. 9:279–
284.
Berrang, M. E., J. F. Frank, R. J. Burh, J. S. Bailey, N. A. Cox,
and J. Mauldin. 1998. Eggshell characteristics and penetration by
Salmonella through the productive life of a broiler breeder flock.
Poult. Sci. 77:1446–1450.
Bialka, K. L., A. Demirci, S. J. Knabel, P. H. Patterson, and V.
M. Puri. 2004. Efficacy of electrolyzed oxidizing water for the
microbial safety and quality of eggs. Poult. Sci. 83:2071–2078.
Caner, C., and M. Yuceer. 2015. Efficacy of various protein-based
coating on enhancing the shelf life of fresh eggs during storage.
Poult. Sci. 94:1665–1677.
Cao, W., Z. Zhu, Z. Shi, C. Wang, and B. Li. 2009. Efficiency of
slightly acidic electrolyzed water for inactivation of Salmonella
enteritidis and its contaminated shell eggs. Int. J. Food Microbiol.
130:88–93.
Chousalkar, K. K., P. Flynn, M. Sutherland, J. R. Roberts, and
B. F. Cheetham. 2010. Recovery of Salmonella and Escherichia
coli from commercial egg shells and effect of translucency on
bacterial penetration in eggs. Int. J. Food Microbiol. 142:207–
213.
Cox, N. A., J. S. Bailey, and M. E. Berrang. 1998. Bactericidal
treatment of hatching eggs I. chemical immersion treatments and
Salmonella. J. Appl. Poult. Res. 7:347–350.
Cox, N. A., M. E. Berrang, and J. A. Cason. 2000. Salmonella pen-
etration of egg shells and proliferation in broiler hatching eggs–a
review. Poult. Sci. 79:1571–1574.
De Reu, K., K. Grijspeerdt, L. Herman, M. Heyndrickx, M. Uytten-
daele, J. Debevere, F. F. Putirulan, and N. M. Bolder. 2006. The
effect of a commercial UV disinfection system on the bacterial
load of shell eggs. Lett. Appl. Microbiol. 42:144–148
De Reu, K., W. Messens, M. Heyndrickx, T. B. Rodenburg, M. Uyt-
tendaele, and L. Herman. 2008. Bacterial contamination of table
eggs and the influence of housing systems. Worlds Poult. Sci. J.
64:5–19.
Duan, D., H. Wang, S. Xue, M. Li, and X. Xu. 2017. Application
of disinfectant sprays after chilling to reduce the initial microbial
load and extend the shelf-life of chilled chicken carcasses. Food
Control 75:70–77.
Guentzel, J. L., K. L. Lam, M. A. Callan, S. A. Emmons, and V.
L. Dunham. 2008. Reduction of bacteria on spinach, lettuce, and
surfaces in food service areas using neutral electrolyzed oxidizing
water. Food Microbiol. 25:36–41.
Haugh, R. R. 1937. The Haugh unit for measuring egg quality. US
Poult. Mag. 43:552–573.
Himathongkham, S., H. P. Riemann, and R. A. Ernst. 1999. Effi-
cacy of disinfection of shell eggs externally contaminated with
Salmonella enteritidis. Int. J. Food Microbiol. 49:161–167.
Holck, A. L., K. H. Liland, S. M. Dromtorp, M. Carlehog, and A.
Mcleod. 2018. Comparison of UV-C and pulsed UV light treat-
ments for reduction of Salmonella, Listeria monocytogenes, and
enterohemorrhagic Escherichia coli on eggs. J. Food Prot. 81:
6–16.
5939
Huang, Y. R., Y. C. Hung, S. Y. Hsu, Y. W. Huang, and D. F.
Hwang. 2008. Application of electrolyzed water in the food in-
dustry. Food Control 19:329–345.
Jin, T. Z., J. B. Gurtler, and S. Q. Li. 2013. Development of an-
timicrobial coatings for improving the microbiological safety and
quality of shell eggs. J. Food Prot. 76:779–785.
Koide, S., J. Takeda, J. Shi, H. Shono, and G. G. Atungulu. 2009.
Disinfection efficacy of slightly acidic electrolyzed water on fresh
cut cabbage. Food Control 20:294–297.
Mansur, A. R., and D. Oh. 2015. Combined effects of thermosonica-
tion and slightly acidic electrolyzed water on the microbial qual-
ity and shelf life extension of fresh-cut kale during refrigeration
storage. Food Microbiol. 51:154–162.
Messens, W., K. Grijspeerdt, and L. Herman. 2005. Eggshell pen-
etration by Salmonella: a review. Worlds Poult. Sci. J. 61:71–
86.
Morsy, M. K., A. M. Sharoba, H. H. Khalaf, H. H. Eltanahy, and C.
N. Cutter. 2015. Efficacy of antimicrobial pullulan-based coating
to improve internal quality and shelf-life of chicken eggs during
storage. J. Food Sci. 80:1066–1074.
Ni, L., W. Cao, W. Zheng, H. Chen, and B. Li. 2014. Efficacy
of slightly acidic electrolyzed water for reduction of foodborne
pathogens and natural microflora on shell eggs. Food Sci. Tech-
nol. Res. 20:93–100.
Rahman, S. M. E., J. Wang, and D. H. Oh. 2013. Synergistic effect
of low concentration electrolyzed water and calcium lactate to
ensure microbial safety, shelf life and sensory quality of fresh pork.
Food Control 30:176–183.
Samli, H. E., A. Agma, and N. Senkoylu. 2005. Effects of storage
time and temperature on egg quality in old laying hens. J. Appl.
Poult. Res. 14:548–553.
Sheng, X. W., D. Shu, X. Tang, and Y. T. Zang. 2018. Effects of
slightly acidic electrolyzed water on the microbial quality and
shelf life extension of beef during refrigeration. Food Sci. Nutr.
6:1975–1981.
Yimenu, S. M., J. Kim, J. Koo, and B. Kim. 2017. Predictive mod-
eling for monitoring egg freshness during variable temperature
storage conditions. Poult. Sci. 96:2811–2819.
Yoshifumi, H. 2003. Improvement of the electrolysis equipment and
application of slightly acidic electrolyzed water for dairy farming.
J. Jpn. Soc. Agric. 65:27–29.
Yuceer, M., M. S. Aday, and C. Caner. 2016. Ozone treatment of
shell eggs to preserve functional quality and enhance shelf life
during storage. J. Sci. Food Agric. 96:2755–2763.
Zang, Y. T., B. M. Li, Sh. Bing, and W. Cao. 2015. Model-
ing disinfection of plastic poultry transport cages inoculated
with Salmonella enteritids by slightly acidic electrolyzed wa-
ter using response surface methodology. Poult. Sci. 94:2059–
2065.
Zang, Y. T., B. M. Li, Z. X. Shi, X. W. Sheng, H. X. Wu, and D.
Q. Shu. 2017. Inactivation efficiency of slightly acidic electrolyzed
water against microbes on facility surfaces in a disinfection chan-
nel. Int. J. Agr. Biol. Eng. 10:23–30.