Food Chemistry 141 (2013) 954–960

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Effect of alginate/nano-Ag coating on microbial and physicochemical

characteristics of shiitake mushroom (Lentinus edodes) during cold
storage
Tianjia Jiang, Lifang Feng, Yanbo Wang ⇑
College of Food Science and Biotechnology, Zhejiang Gongshang University, Food Safety Key Lab of Zhejiang Province, Hangzhou 310035, PR China

a r t i c l e i n f o a b s t r a c t

Article history: The effect of a novel alginate/nano-Ag coating material on the preservation quality of shiitake mushroom
Received 3 December 2012 (Lentinus edodes) during 4 ± 1 °C storage was investigated. The results showed that the alginate/nano-Ag
Received in revised form 22 February 2013 coating had quite a beneficial effect on the physicochemical and sensory quality, compared to the control
Accepted 26 March 2013
treatment. After a 16-day storage, mushroom weight loss, softening, and browning of the alginate/nano-
Available online 15 April 2013
Ag coating were significantly inhibited. The lower microbial counts, including mesophilic, psychrophilic,
pseudomonad, and yeasts and moulds, in treated mushrooms during storage should be attributed to the
Keywords:
alginate/nano-Ag coating. Meanwhile, the contents of the reducing sugar, total sugar, total soluble solids
Shiitake mushroom
Alginate/nano-Ag
and electrolyte leakage rate were increased to 3.9 mg/g, 11.2 mg/g, 5.1% and 16.5% for the alginate/nano-
Microbiological quality Ag coating and 3.7 mg/g, 8.3 mg/g, 6.3% and 31.7% for the control treatment. Therefore, the alginate/nano-
Physicochemical property Ag coating could be applied for preservation of the shiitake mushroom to expand its shelf life and
Storage life improve its preservation quality.
Ó 2013 Elsevier Ltd. All rights reserved.

1. Introduction to increase film flexibility and processability and oxygen perme-

ability. Alginate is a GRAS (generally recognised as safe) substance
The shiitake mushroom (Lentinus edodes) is one of the most (FDA), and it has been used to reduce weight loss and natural
common edible mushrooms, traditionally cultivated in China. It microflora counts in carrots (Amanatidou, Slump, Gorris, & Smid,
is also the second most cultivated edible mushroom, comprising 2000), and also to maintain the quality and prolong shelf life of
25.4% of the worldwide production (Boa, 2004). Shiitake mush- fresh-cut apples (Rojas-Graü et al., 2007). In addition, this type of
rooms are highly perishable and tend to lose quality right after har- film has shown potential as a carrier of active or functional food
vest. Their shelf life is short because of their high respiration rate, ingredients such as antibrowning agents, colourants, flavours,
tendency to turn brown and having no physical protection to avoid nutrients, spices, and antimicrobial compounds that can extend
water loss or a microbial attack. Bacteria, moulds, enzymatic activ- the product shelf-life and reduce the risk of pathogen growth on
ity and biochemical changes can cause spoilage during storage. food surfaces (Lin and Zhao, 2007; Rojas-Graü, Soliva-Fortuny, &
Gram-negative microorganisms, such as Pseudomonas tolaasii, Martín-Belloso, 2009). Incorporating antimicrobial compounds
Pseudomonas fluorescens and yeasts, such as Candida sake, have into edible films or coatings provides a novel way to improve the
been associated with mushroom spoilage (Masson, Ainsworth, Ful- safety and shelf-life of fruits and vegetables. Some of the more
ler, Bozkurt, & Ibanoglu, 2002). The short shelf-life of a mushroom commonly used antimicrobials include benzoic acid, sorbic acid,
is an impediment to the distribution and marketing of the fresh lysozyme, lactoferrin, bacteriocins (nisin and pediocin) and plant-
product. derived secondary metabolites, such as phytoalexins and essential
Alginate, a polysaccharide derived from the marine brown al- oils.
gae, has been preponderant in making edible films, due to its un- In recent years, nanomaterials have attracted increasing atten-
ique colloidal properties and its ability to form strong gels or tion because of their potential impact on a wide range of industries
insoluble polymers upon reaction with multivalent metal cations and markets (Michael, 2004; Guz, Rodger, Guz, Guz, 2007). Among
such as calcium (Rhim, 2004). Plasticisers, such as glycerol, are noble-metal nanomaterials, nano-silver (NS) has received consid-
usually required for polysaccharide- and protein-based edible films erable attention due to its attractive physicochemical properties.
It is well known that silver, in various chemical forms, is toxic to
⇑ Corresponding author. Tel./fax: +86 571 88071024x7589. a wide range of microorganisms (Liau, Read, Pugh, Furr, Russell,
E-mail address: wang58516@sohu.com (Y. Wang). 1997). Indeed, NS has been shown to be a promising antimicrobial

0308-8146/$ - see front matter Ó 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.foodchem.2013.03.093
 T. Jiang et al. / Food Chemistry 141 (2013) 954–960 955

material (Sondi and Salopek-Sondi, 2004). The larger surface area (both at 20 °C and 100% RH) and 2.8  105–6.5  105 g m2 s1
of NS can improve its antibacterial effectiveness against 150 types for H2O (at 37 °C and 90% RH), then stored for 16 days at 4 ± 1 °C
of microbes. NS particles can attach to the cell membranes and and 95% relative humidity (RH). Fifteen replicates were included
penetrate into bacteria. Therein, NS can disrupt processes such as in each treatment group, and subsequently every 4 days, three rep-
respiration and cell division, leading to cell death. NS releases sil- licates from each treatment group were analysed.
ver ions (Ag+) within bacterial cells, which enhance its bactericidal
activity (Rai, Yadav, Gade, 2009). In cut flower applications, NS has 2.2. Microbiological analysis
been reported to be effective as an antimicrobial agent, an ethylene
inhibitor (Kim, Lee, Suh, 2005) and/or a regulator of stomatal aper- All samples were analysed for the mesophilic, psychrophilic,
ture (Lv, He, Li, Cao, Xu, 2010). Some researchers have investigated pseudomonad, and yeasts and moulds counts. Twenty-five grams
the safety of solutions containing small concentrations of the silver of mushrooms were removed aseptically from each pack and
ion (USEPA, 2001). The US Environmental Protection Agency diluted with 225 ml of 0.1% peptone water. The samples were
(USEPA) proposed that the secondary maximum contaminant level homogenised by a stomacher at a high speed for 2 min. Serial dilu-
of silver ion in drinking water must be less than 0.10 mg/l (USEPA, tions (101–109) were made in serial dilution tubes by taking
2001). 1.0 ml with 9.0 ml of 0.1% peptone water. Aerobic counts were
However, to the best of our knowledge, the use of alginate as a determined on a plate count agar (PCA; Merck) following incuba-
natural antimicrobial agent, either individually or in combination tion at 35 °C for 2 days for mesophilic bacteria, and at 4 °C for
with nanomaterials, including NS, has not been studied to date, 7 days for psychrophilic bacteria. The microorganism, Pseudomo-
in fresh shiitake mushrooms. Thus, the objective of the present nas, was counted on cephaloridin fucidin cetrimide agar (CFC;
study was to determine the effect of the alginate/nano-Ag coating Difco), with a selective supplement SR 103 (Oxoid). The incubation
on the microbial and physicochemical characteristics of shiitake temperature was 25 °C and plates were examined after 48 h. Yeasts
mushrooms, during storage at 4 °C for 16 days. and moulds were estimated on a potato dextrose agar (PDA;
Merck) and incubation conditions were of 28 ± 1 °C for 5–7 days.

2. Materials and methods

2.3. Package atmosphere composition
2.1. Preparation of coating solutions and samples
O2 and CO2 concentrations in packages were evaluated by using
a SCY-2A O2 and CO2 analyzer (Xinrui Instrument Co., Shanghai,
Food-grade alginate was purchased from Shanghai Kaiyang Bio-
China). Gas samples were taken from the bags with a 20 ml
technology Co., Ltd. (Shanghai, China). Silver nitrate (AgNO3) and
syringe.
sodium borohydride (NaBH4) were purchased from Shanghai
Chemical Reagent Co., Ltd. (Shanghai, China). Alginate concentra-
tions were selected on the basis of initial screening experiments. 2.4. Weight loss and firmness evaluation
Sodium alginate solution (1.5%, w/v) was prepared by mixing
7.5 g of alginate with 500 ml of distilled water and stirred at a con- Weight loss was determined by weighing the whole mushroom
trolled temperature of 70 °C, until the mixture became clear. before and after the storage period. Weight loss was expressed as
AgNO3, NaBH4 and polyvinylpyrrolidone (PVP) were dissolved in the percentage of loss of weight with respect to the initial weight.
deionised water to form an aqueous solution of AgNO3 (0.1 M), A penetration test was performed on the shiitake mushroom cap
NaBH4 (0.01 M) and PVP (0.01 M), respectively. The aqueous solu- using a TA.XT Express-v3.1 texture analyzer (Stable Micro Systems,
tions of PVP and NaBH4 were mixed at a volume ratio of 1:1. About UK), using a 5 mm diameter cylindrical probe. Samples were pen-
500 ml of this solution was transferred to a beaker, 7.5 g of alginate etrated 5 mm in depth. The speed of the probe was 2.0 mm s1
was added, and the solution agitated with a magnetic stirrer before during the pretest, as well as during penetration. Force and time
adding the AgNO3 solution. Upon addition of silver nitrate drop by data were recorded with Texture Expert (Version 1.0) from Stable
drop, the colourless solution of NaBH4–PVP slowly changed from Micro Systems. From the force vs. time curves, firmness was de-
yellow to pale brown, indicating the formation of an alginate/ fined as the maximum force.
nano-Ag composite. Glycerol (3%, w/w) was mixed with the pre-
pared alginate/nano-Ag solution as plasticiser and stirred thor- 2.5. Reducing sugar, total sugar, total soluble solids and electrolyte
oughly. A 2% (w/v) calcium chloride solution was prepared for leakage rate
cross-linking. The final practical silver concentration at 0.06 mg/l
for coating was used, which is safe to consume according to USEPA The reducing sugar and total sugar in mushrooms was deter-
(2001). mined according to the methods of Miller (1959) and Dubois, Gil-
The shiitake mushrooms used in this study, were harvested les, and Hamilton (1956), respectively. The mushroom was ground
from a local farm in Hangzhou, China. The mushrooms were trans- in a mortar and squeezed with a hand press, and the juice was ana-
ported to the laboratory within 1 h of picking, then stored in dark- lysed for the presence of total soluble solids (TSS). The TSS was
ness at 4 ± 1 °C and 90% relative humidity (RH) for 12 h. The measured at 25 °C with a refractometer (Taiguang 405225, Taihua
mushrooms were divided into three samples of 60 each. Three dif- Optical Co., Ltd., Chengdu, China). The electrolyte leakage rate was
ferent treatments were used: (1) control; (2) alginate coating, and measured essentially as described by Autio and Bramlage (1986).
(3) alginate/nano-Ag coating. The mushrooms were dipped into the Mushroom fruit bodies (5 g) were cut into four pieces, leaving
solution for 2 min. Samples dipped in distilled water were used as the pileus intact, washed three times with deionised water to re-
the control. Treated samples were kept over a plastic sieve for move surface-adhered electrolytes. After drying with a filter paper,
20 min and a fan generating low-speed air, was used to hasten dry- they were suspended in 40 ml of deionised water in a 100 ml bea-
ing. Then, a tissue paper was used to absorb any excess solution ker and shaken at 25 °C on a rotary shaker for 30 min; The electri-
from the surface. The treated samples were put and sealed in cal conductivity was measured immediately (P0) and again after
18 cm  20 cm bags of low density polyethylene (PE) (0.04 mm 10 min (P1). Samples were then boiled for 10 min and cooled to
thickness), the PE gas transmission rates were 1078  1018 room temperature, and a final conductivity measurement (P2)
mol m1 s1 Pa1 for O2, 4134  1018 mol m1 s1 Pa1 for CO2 was taken. The relative electrolyte leakage rate (RELT) was
956 T. Jiang et al. / Food Chemistry 141 (2013) 954–960

calculated according to the following equation: (P1  P0)/(P2  P0) nated during storage in all of the analysed samples. In all of the
and expressed as a percentage. studied treatments, the psychrophilic bacteria counts increased
less than two order during the entire storage period. All the sam-
2.6. Sensory evaluation ples had counts below 104 cfug1. This contamination level sug-
gests that shiitake mushrooms in the studied coating conditions
The sensory attributes that characterised mushroom deteriora- did not favour the development of this type of bacteria. Mush-
tion were determined. These attributes were: off-odour, gill colour, rooms from the control treatment exhibited tiny brown spots on
gill uniformity, cap surface uniformity, and presence of dark zones day 4 that developed into dark zones, characteristic of Pseudomo-
on the cap (Ares, Parentelli, Gámbaro, Lareo, Lema, 2006). Samples nas spoilage by day 8. The mushrooms were highly decayed at this
were evaluated by a sensory panel of ten trained assessors. The point and the end of their shelf-life was due to microbial spoilage.
mushrooms were served in closed, odourless plastic containers at The alginate/nano-Ag coated samples did not exhibit these charac-
room temperature. After opening polyethylene bags, the mush- teristics of microbial degradation even on day 12. The organisms
rooms were placed in plastic containers and evaluations were per- usually responsible for spoilage of mushrooms are Gram-negative,
formed within 2 h in order to avoid loss of off-odours. A balanced psychrotrophic bacteria, particularly belonging to the Pseudomona-
complete block design was carried out for a duplicate evaluation ceae family, due to contamination of the product from compost.
of the samples. For scoring, 10 cm unstructured scales anchored Sanpui, Murugadoss, Prasad, Ghosh, and Chattopadhyay (2008)
with ‘‘nil’’ for zero and ‘‘high’’ for ten were used, except for the gill found that the chitosan–Ag–nanoparticle composite has a signifi-
colour descriptor, for which the anchors were ‘‘white’’ and cantly higher antimicrobial activity than its components, at their
‘‘brown’’. respective concentrations. The presence of a small percentage
(2.15%, w/w) of metal nanoparticles in the composite was enough
to significantly enhance inactivation of Escherichia coli as compared
2.7. Statistical analysis
with unaltered chitosan. Similar results were found in the present
study, and the presence of a silver nanoparticle incorporated by
Experiments were performed using a completely randomised
alginate coating seems to have contributed to a certain decrease
design, and each was composed of three replicates. Data were sub-
in the microbial counts of shiitake mushrooms, improving the gen-
jected to a one-way analysis of variance (ANOVA). Mean separa-
eral quality after storage.
tions were performed by Tukey’s multiple range test (DPS
version 6.55). Differences at P < 0.05 were considered significant.
3.2. Effect of alginate/nano-Ag coating on gas composition
3. Results and discussion
Similar patterns of CO2 and O2 levels within the packaging were
3.1. Effect of alginate/nano-Ag coating on microbiological quality found for all the treatments (Table 1). The equilibrium-modified
atmosphere (steady state) was attained within the packaging after
It is evident from this study that alginate/nano-Ag coating was 4 days. Mushrooms subjected to control treatment, showed
more effective in reducing microbial counts than other treatments slightly higher CO2 and lower O2 concentrations within the packag-
(Fig. 1). Mesophilic, pseudomonad, yeasts and moulds predomi- ing than alginate and alginate/nano-Ag coating, indicating that

A 7
B5
6
4
Mesophilic count

psychrophilic count

5
(log10 cfu/g)

(log10 cfu/g)

4 3
3
Control 2
2 Alginate
1 Alginate/nano-Ag 1
0
0
0 4 8 12 16
0 4 8 12 16
Storage time (days)
Storage time (days)

9
C 8 D9
8
Pseudomonad count

7
Yeasts and molds count

7
(log10 cfu/g)

6
6
(log10 cfu/g)

5
5
4
4
3 3
2
2
1 1
0 0
0 4 8 12 16 0 4 8 12 16
Storage time (days) Storage time (days)

Fig. 1. Effect of alginate/nano-Ag coating on mesophilic (A), psychrophilic (B), pseudomonad (C), and yeasts and moulds (D) counts (log10 cfug1) change of shiitake
mushrooms stored at 4 °C for 16 days. Each data point is the mean of three replicate samples. Vertical bars represent standard errors of means.
 T. Jiang et al. / Food Chemistry 141 (2013) 954–960 957

Table 1 Ag. During the steady state, the mushroom respiration (O2 con-
Effect of alginate/nano-Ag coating coating on gas composition (%) changes of shiitake sumption and CO2 production) was balanced by O2 and CO2 diffu-
mushrooms stored at 4 °C for 16 days.A,B
sion through the film: the O2 and CO2 concentrations reached
Days at 4 °C Control Alginate Alginate/nano-Ag values of 17.5% and 3.5% respectively in alginate/nano-Ag coat-
O2 ing. In the present study, coating shiitake mushroom with alginate
0 21 21 21 or alginate/nano-Ag showed a similar reduction because 2% cal-
1 13.69 ± 0.24c 14.28 ± 0.13b 15.05 ± 0.31a cium chloride was added. The calcium ion is known to inhibit
2 14.40 ± 0.14c 15.65 ± 0.16b 16.81 ± 0.19a
3 15.36 ± 0.28c 16.55 ± 0.05b 17.49 ± 0.25a
respiratory activity, as calcium ions are involved in the function
4 15.89 ± 0.32c 16.80 ± 0.14b 18.32 ± 0.47a of many enzymes actions in cellular and physiological processes
8 14.48 ± 0.15c 15.62 ± 0.28b 17.20 ± 0.22a (Oms-Oliu, Soliva-Fortuny, Martín-Belloso, 2008). Lee, Park, Lee,
12 14.29 ± 0.38c 16.34 ± 0.22b 17.37 ± 0.10a and Choi (2003) suggested that the large reduction of initial respi-
16 15.10 ± 0.32c 16.72 ± 0.37b 17.62 ± 0.32a
ration rate in whey protein concentrate (WPC)-coated apples is the
CO2 effect of calcium ion present in the aqueous WPC solution.
0 0 0 0
1 10.13 ± 0.14a 8.78 ± 0.26b 7.67 ± 0.09c
2 6.77 ± 0.13a 5.85 ± 0.14b 5.05 ± 0.25c 3.3. Effect of alginate/nano-Ag coating on weight loss and firmness
3 5.63 ± 0.07a 4.70 ± 0.13b 3.98 ± 0.19c
4 5.25 ± 0.04a 4.21 ± 0.20b 3.02 ± 0.13c
Mushroom weight loss is mainly caused by the water transpira-
8 5.95 ± 0.13a 4.80 ± 0.04b 3.31 ± 0.39c
12 5.10 ± 0.26a 4.38 ± 0.07b 3.86 ± 0.20c tion rate and CO2 loss during respiration. The thin skin of shiitake
16 4.73 ± 0.19a 3.89 ± 0.14b 3.14 ± 0.14c mushrooms makes them susceptible to rapid water loss, resulting
A in shrivelling and deterioration. As shown in Fig. 2A, the highest
Mean of three replications ± standard deviation.
B
Mean values in the same row with different letters are significantly different weight loss was observed in the control samples, where it reached
(P < 0.05). 4.1% at the end of storage. From the viewpoint of Bico, Raposo,
Morais, and Morais (2009) more than 4–6% (of the total fresh
weight) weight-loss was accompanied by visible wilting or wrin-
control samples have a higher rate of respiration. Mushrooms kling of the surface of the fruits and vegetables. For all of the coated
coated with alginate and alginate/nano-Ag showed lower CO2 level mushrooms, the weight-loss was less than 4%, which indicates that
within the packaging. It was obvious that the respiration rate after the coated mushroom maintained freshness during storage. This
4 days of storage at 4 °C, was inhibited by alginate coating treat- was ascribed to the fact that edible coatings act as barriers, thereby
ment and the respiration rate decreased with addition of nano- restricting water transfer and protecting mushroom epidermis

5
A
4.5
4

3.5 Control
Weight loss (%)

3 Alginate
2.5 Alginate/nano-Ag
2
1.5

1
0.5
0

Storage time (days)

4
B
3.5

2.5 Control
Firmness (N)

2 Alginate
1.5 Alginate/nano-Ag

0.5

0
0 4 8 12 16
Storage time (days)

Fig. 2. Effect of alginate/nano-Ag coating on weight loss (A) and firmness (B) change of shiitake mushrooms stored at 4 °C for 16 days. Each data point is the mean of three
replicate samples. Vertical bars represent standard errors of means.
958 T. Jiang et al. / Food Chemistry 141 (2013) 954–960

from mechanical injuries, as well as sealing small wounds and thus 3.4. Effect of alginate/nano-Ag coating on reducing sugar, total sugar,
delaying dehydration. The largest weight loss reduction was ob- total soluble solids and electrolyte leakage rate
tained from the coated application of alginate/nano-Ag and algi-
nate during storage time. Reducing sugar levels increased in all samples during the 16-
Loss of firmness is one of the main factors limiting quality and day storage period (Fig. 3A). Although the rate of increase was ini-
the postharvest shelf-life of the shiitake mushroom. The general tially generally constant in alginate, alginate/nano-Ag and control,
trend in firmness changes in mushroom in all treatments stored levels increased at an accelerated rate between 4 and 16 days post-
at 4 °C is shown in Fig. 2B. The coating significantly improved harvest in samples from alginate coating treatment. A smaller
firmness of the mushroom. Incorporation of a silver nanoparticle acceleration was observed between 8 and 16 days postharvest in
into alginate had a significant and positive effect on the firmness. control and alginate/nano-Ag coating. The total increase in reduc-
Indeed, samples coated with alginate/nano-Ag maintained their ing sugar in alginate coated samples was 37.1%. In contrast, rela-
firmness during the entire storage period. Softening can occur be- tively small increases in the reducing sugar content of alginate/
cause of the degradation of cell walls in postharvest mushrooms nano-Ag coated and control samples were recorded. Levels re-
by bacterial enzymes and increased activity of endogenous autol- mained relatively steady throughout the postharvest period and
ysins (Zivanovic, Buescher, Kim, 2000). Microorganisms such as were only 19.4% higher than initial levels when the experiment
Pseudomonas degrade mushrooms by breaking down the intracel- was terminated, suggesting that the mushroom with alginate/
lular matrix and reducing the central vacuole, resulting in par- nano-Ag coating and control treatments synthesised reducing sug-
tially collapsed cells and a loss of turgor. This kind of bacterial- ars at a lower rate than did the alginate coating. These results indi-
induced softening was observed in control samples but was cated that the application of alginate/nano-Ag coating might be
inhibited by the alginate/nano-Ag coating treatment due to its able to slow the metabolism to give prolonged storage life to the
higher antibacterial activity. On the other hand, the use of cal- mushroom.
cium chloride to cross-link the polymer matrix, could also repre- The total and soluble sugar concentrations in harvested plant
sent a beneficial effect for the coated mushroom by delaying its products are considered important indicators of postharvest dete-
softening. Similar results were obtained by Lee et al. (2003), rioration (Hammond and Nichols, 1975). Total sugar levels in algi-
who found that the incorporation of 1% calcium chloride within nate coated samples increased at a rate significantly different
a whey protein formulation helped to maintain firmness of (P < 0.05) from the control over the first 12-day period, and
fresh-cut apple slices. Firming and resistance to softening result- 155.0% higher than initial concentration at the end of the experi-
ing from addition of Ca2+ have been attributed to the stabilisation ment (Fig. 3B). However, there was a much lower increase in the
of membrane systems and the formation of Ca pectates, which in- sugar content of control mushrooms during the whole storage,
crease the rigidity of the middle lamella and cell wall (Jackman which was 3.8% of the initial carbohydrate as compared to 40.0–
and Stanley, 1995). 55.0% in the other treatments. In contrast, steady decreases in

A 5 B 18
16
Reducing sugar (mg/g FW)

Total sugar (mg/g FW)

4.5
14
4 12
10
3.5 8
3 Control
6
Alginate
4
2.5 2
Alginate/nano-Ag
2 0
0 4 8 12 16 0 4 8 12 16

Storage time (days) Storage time (days)

C9
D 40
Soluble solids concentration (%)

8
35
Relative leakage rate (%)

7
30
6
5 25

4 20
3 15
2 10
1 5
0 0
0 4 8 12 16 0 4 8 12 16
Storage time (days) Storage time (days)

Fig. 3. Effect of alginate/nano-Ag coating on reducing sugar (A), total sugar (B), total soluble solids (C), and electrolyte leakage rate (D) change of shiitake mushrooms stored
at 4 °C for 16 days. Each data point is the mean of three replicate samples. Vertical bars represent standard errors of means.
 T. Jiang et al. / Food Chemistry 141 (2013) 954–960 959

the total sugar content were reported in Agaricus bisporus mush- alginate/nano-Ag coated mushrooms had a significantly lower rel-
rooms stored at 12 °C for 12 days (Tseng and Mau, 1999). ative leakage rate than the control mushrooms, indicating that a
The total soluble solids (TSS) of the shiitake mushroom in- higher membrane integrity was maintained.
creased with time during 4 °C storage (Fig. 3C). The TSS of control
mushrooms increased after 4 days of storage whilst alginate/nano- 3.5. Effect of alginate/nano-Ag coating on sensory attributes
Ag coated mushrooms experienced a slight increase during the
same period. Similar results were obtained by Tao, Zhang, Yu, The average values for the sensory attributes of coated and con-
and Sun, 2006, who reported an increase in the TSS in button trol shiitake mushrooms, during 16 days storage, are shown in Ta-
mushrooms stored at 4 ± 1 °C and 75% RH. It can be expected that ble 2. As expected, mushroom off-odour, gill colour, gill uniformity,
TSS increases during mushroom senescence and decreases in cap uniformity and dark zones significantly (P < 0.05) changed
senescent mushroom due to respiration. A plausible explanation with storage time, supporting the validity of using these parame-
for the observed increment in the TSS is the considerable loss of ters as indicators of mushroom deterioration. The off-odour inten-
water suffered by mushrooms during storage. Indeed, the greater sity significantly increased after 8 d of storage in control samples.
changes in TSS occurred in those mushrooms which suffered the The colour of mushroom gills gradually became browner and less
greatest water loss. The solubilisation of the cell wall polyuronides uniform with time for all the evaluated conditions. The gills of con-
and hemicelluloses in senescent mushroom might also contribute trol mushrooms showed a colour intensity of 5.38 and uniformity
to the increase in TSS. In addition, decreased respiration rates also of 5.10 at the 12th day of storage. However, the gills of alginate/
slow down the synthesis and use of metabolites resulting in lower nano-Ag coated mushrooms did not reach these intensities even
TSS, due to the slower hydrolysis of carbohydrates to sugars (Ya- at the end of storage. A better trend was observed for the unifor-
man and Bayoindirli, 2002). mity of the cap surface and for the presence of dark stains on the
The changes in electrolyte leakage of mushrooms are presented cap in alginate/nano-Ag samples. The browning of mushrooms is
in Fig. 3D. The initial electrolyte leakage was 1.7% in the shiitake attributed to the oxidation of phenolic compounds, the action of
mushrooms. In general, electrolyte leakage was enhanced as the polyphenol oxidase (PPO) enzyme and the action of bacteria and
storage time increased. The electrolyte leakage contents of algi- mould on the mushroom tissue. As alginate/nano-Ag causes coat-
nate/nano-Ag coated samples and control samples were 16.5% ing reduction of spoilage organisms, such as Pseudomonas, respon-
and 31.7%, respectively, at the end of storage. The increase of mem- sible for the oxidation of phenolic compounds to form brown-
brane permeability in the control groups was much higher than coloured melanins, it prevents the formation of brown patches,
that in the alginate/nano-Ag coatted mushrooms. These results hence improving the appearance and colour of the mushrooms.
demonstrated that membrane permeability, as an indicator of These results suggest that the alginate/nano-Ag coating treatment
membrane integrity, gradually increased during storage. However, was more effective in retarding mushroom sensory deterioration.

4. Conclusions
Table 2
Effect of alginate/nano-Ag coating on sensory attributes change of shiitake mush- In this study, a novel alginate/nano-Ag coating material was ap-
rooms stored at 4 °C for 16 days.A,B
plied to the preservation of shiitake mushroom during 4 ± 1 °C
Days at 4 °C Control Alginate Alginate/nano-Ag storage. The results showed that the alginate/nano-Ag coating
Off-odour had quite beneficial effects on the physicochemical and physiolog-
0 0 0 0 ical quality compared to the control treatment. Therefore, the algi-
4 1.32 ± 0.04a 1.22 ± 0.03b 1.28 ± 0.03ab nate/nano-Ag coating may provide an attractive alternative, to
8 2.76 ± 0.13a 2.53 ± 0.12b 2.21 ± 0.13c
improve the preservation qualities of the shiitake mushroom dur-
12 5.20 ± 0.19a 4.86 ± 0.18b 3.52 ± 0.15c
16 6.62 ± 0.20a 5.70 ± 0.16b 5.18 ± 0.24c ing an extended storage. Moreover, further research will be needed
to explore the exact alginate/nano-Ag coating mechanism during
Gills colour
0 0 0 0 storage, to facilitate the application of nanotechnology over a
4 1.43 ± 0.11a 1.40 ± 0.05a 1.14 ± 0.07b broader range in the future.
8 2.94 ± 0.17a 2.65 ± 0.16b 2.21 ± 0.10c
12 5.38 ± 0.31a 4.78 ± 0.25b 3.40 ± 0.16c
16 6.87 ± 0.28a 5.83 ± 0.14b 4.42 ± 0.21c Acknowledgment
Gills uniformity
0 10 10 10 This study was supported by the Ph.D. Programs Foundation of
4 8.73 ± 0.24b 8.64 ± 0.14c 8.80 ± 0.29a Ministry of Education of China (No. 20123326120003).
8 6.22 ± 0.21c 6.53 ± 0.26b 6.73 ± 0.18a
12 5.10 ± 0.16c 5.58 ± 0.24b 5.80 ± 0.26a
16 4.21 ± 0.18b 4.43 ± 0.14b 4.92 ± 0.23a
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