Chitosan Coating Effect on Storability

of Fresh Strawberries
AHMED

EL GHAOUTH,

JOSEPH AWL,

RATHY PONNAMPALAM,

ABSTRACT

The effect of chitosan coating (1.0 and 1.5% w/v) in controlling decay
of strawberries at 13C was investigated as compared to a fungicide,
iprodione (Rovralm). Chitosan coating significantly reduced decay of
berries (PsO.05) compared to the control. There was no significant
difference between chitosan and fungicide treatments up to 21 days
storage. Thereafter, Rovrala-treated berries decayed at a higher rate
than chitosan-coated berries. Chitosan-coated berries stored at 4C
were firmer, higher in titratable acidity, and synthesized anthocyanin
at a slower rate than Rovrala-treated or nontreated berries. Chitosan
coating decreasedrespiration rate of the berries with a greater effect
at higher concentration.

and Quality
and MARCEL

BOULET

phytoalexin (pisatin) in pea pods (Hadwiger and Beckman,

1980; Kendra and Hadwiger, 1984). Due to its ability to form
semi-permeable film (Bai et al., 1988), chitosan coating can
be expected to modify the internal atmosphere as well as decrease the transpiration losses. Therefore a delay in ripening
and control of decay by means of chitosan coating could result.
Chitosan is nontoxic (Arai et al., 1968), and its biological
safety has been recently demonstrated by feeding trials with
domestic animals (Hirano et al., 1990). The objective of our
study was to assess the potential of chitosan coating as an
antifungal agent compared to a fungicide, iprodione (Rovralm),
in controlling decay of postharvest strawberries, and to determine the effect of chitosan coating on quality and storability.

INTRODUCTION
STRAWBERRY FRUIT (Fragatiu ananussu) is highly perishable and its storage life is often terminated by fungal infection causedby Botrytis cinereu and Rhizopus sp. (Maas, 1981).
The most prevalent method of maintaining quality and controlling decay of strawberries is by rapid cooling after harvest
and storage at low temperatures, typically 1C with high humidity. Since effective control of temperature during transit
and storage of strawberries is difficult, other means of preservation have been sought. Because strawberries can tolerate
elevated CO, atmosphere, they are transported in pallet-bags
under high COZ (Bell, 1986). Although high CO* controls decay (El Kazzaz et al., 1983), prolonged exposure to CO1 can
cause development of off-flavors (Woodward and Topping,
1972).
Postharvest decay of strawberries can also be controlled by
application of fungicides (Jordan, 1973; Aharani and Barkai-

Golan, 1987). However, fungicides leave residues and the

number of fungicide-tolerant postharvest pathogens is growing
(Spotts and Cervantes, 1986). Thus, efforts have been made
to replace fungicides by natural products or to intensify natural
defensesof the tissue to control decay and prolong storage life
(Adikaram et al., 1988; Boulet et al., 1989).
Recently, semi-permeable coatings have been advanced to
improve storability of perishable crops (Lowing and Cutts,
1982). Application of <<Pro-Long>>
(a blend of sucrose esters
of fatty acids and sodium carboxymethyl cellulose) to bananas
delayed ripening by modifying the internal atmospheres(Banks,
1984). Delay of ripening was also reported in pears and apples
coated with c<Nutri-Save%(N,O-carboxymethyl chitosan) (Elson et al., 1985; Davis et al., 1988). However, for strawberries
coating with semi-permeable film has not been explored.
Chitosan, a high molecular weight cationic polysaccharide,
theoretically should be an ideal preservative coating material
for strawberries. It has been shown to inhibit growth of several
fungi (Allan and Hadwiger, 1979; St&se1 and Leuba, 1984;
El Ghaouth et al., 1989; Hirano and Nagao, 1989), to induce
chitinase, a defense enzyme (Mauch et al., 1984) and to elicit
Authors El Ghaouth, Arul, and Ponnampalam are with the Dept.
of Food Science & Technology, and Horticulture Reseach Center, Universitb laval, Sainte-Foy (Quebec) Canada GlK 7P4. Author Boulet is with the Food Research & Development Centre,
Agriculture Canada, Saint-Hyacinthe (Qubbec), Canada J2S 8E3.
Address inquiries to Dr. Arul.

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MATERIALS

& METHODS

Fruits and chemicals

Strawberries (Fragatiu ananns~a Duch. cv Kent) grown in local

farms were harvested and immediately cooled with vapors of dry ice.
The berries of uniform size with 50% or less red color, free of physical
damage and fungal infection were used. The berries were randomly
distributed into groups of 70 fruit. Each group represented one replicate, and for each treatment four replicates were used. Crab-shell

chitosanwas purchasedfrom ICN Inc. (Cleveland,OH). Iprodione,

(Rovral@)was obtained from May and Baker Canada Inc. All other

chemicalswere of analyticalgrade.
Chitosan coating solutions

To prepare 100 mL of chitosan solutions (1.0 and 1.5%) w/v), 1.0

or 1.5g of chitosan was dispersed in 80 mL of sterile deionized water
to which 2.5 mL of 10N HCl was added to dissolve the chitosan. The
pH of the solution was adjusted to 5.6 with 1N NaOH, and 0.1 mL
of Tween 80 was added to the solution to improve wettability. The
solution was then made up to 100 mL. An acid solution containing
Tween 80 without chitosan, pH 5.6, was used as control.
Preparation

of inoculum

Bovtis cinereu was isolated from infected strawberries and maintained on potato dextrose agar (PDA). Conidia of B. cinerea were
recovered by filtering the my&al s&pension of 2-wk old culture
through 3 layers of sterile cheese cloth. The concentration of the
conidial suspensionwas adjusted to 2 x 105 conidia per mL.
Decay control

Strawberries were immersed in a conidial suspension of B. cinereu

containing 0.1% (v/v) Tween 80, and allowed to air dry at room
temperature for 2 h. Inoculated berries were individually dipped in
aqueous suspension of Rovral@ (100 &mL) or in chitosan solutions
(1.0 and 1.5% w/v) or in water control. Four replicates of 70 berries
were used for each treatment. After drying, the berries were stored at
13C in containers continuously ventilated with humidified (95% RH)

air at 20 Whr. Berrieswere examinedfor mold on alternatingdays

and a beny was consideredinfectedwhen a visible lesion was observed. The results were expressed as percentage of berries infected.
The results were subjected to Analysis of Variance (ANOVA) with
5% LSD values calculated to separatesignificantly different means of
the treatments.

60-

2
1
Time ( days )
Fig. 1 -Effect of chitosan coating, 1.0% (o), 1.5% (o), and RovraP
(iprodione) treatment @) on the decay control of strawberries
stored at 13C compared to the control (water-treated) berries
e). Means of four replicates. Vertical bars represent LSD at 5%
level among treatments.

Quality attributes
The effect of chitosan coating and Rovrale on quality was assessed
separatelywith non-inoculatedberries. They were dipped in Rovral@
suspension(100 ug/mL) or in chitosan solutions (1.0 and 1.5% w/v)
or in water control and stored at 4C as described. All treatments
contained 0.1% Tween 80. Each treatment had 4 replicates, and each
replicate consisted of 70 berries. Quality of the berries was assessed
eachweek. A sample of 15-20 berries in total was randomly removed
from each treatment and analyzed for firmness, titratable acidity and
anthocyanin content. To determine firmness, the berries were sliced
into halves and each half was punch tested. The penetration force
(Newton) of the flesh was measuredwith an Instron Universal Testing
Machine (Model 1101, Instron Corp., Canton, MA) using a 4 mm
flat plunger (Holt, 1970). Titratable acidity was expressedas mg of
citric acid/100 mL. Acidity was determinedusing 10 g aliquot of puree
in 40 mL deionized water and titrating with 0.1 N NaOH to an endpoint of pH 8.1. Anthocyanins were extracted with acidified ethanol
from a 2 g aliquot of a homogenateof 7 berries, according to the
method of Fuleki and Francis (1968). Anthocyanin content was expressedas mg anthocyanin/lOOgstrawberry homogenate.The quality
evaluation data were subjected to Analysis of Variance (ANOVA).

Respiration
The respiration rate of strawberries at 4C was determined by placing them (120 g) in an air tight container (1.35-L) for 2 to 4 hr. Then
a 5 mL gas samplewas withdrawn with a gas tight hypodermic syringe
and analyzed for CO* using a gas chromatograph equipped with a
TCD detector and a Poropak N Column. Four replicates of each treatment were analyzed.

RESULTS & DISCUSSION

Antifungal effect of chitosan coating and RovraP
Decay of strawberries was reduced significantly (P~0.05)
when inoculated-berrieswere either coated with chitosan or
dipped in Rovral@(Fig. 1). The early signs of mold development in the strawberries, regardlessof the treatment, appeared
after 8 days storage at 13C. After 21 days at 13C, the percentagesof decayed-berriesin chitosan-coated(1.0 and 1.5%
w/v) and Rovral@-treatedwere 11, 10 and 13% respectively,
while in the control it was 52%. There was no significant
difference between chitosan and Rovral@
treatment in controlling decay before 21 d storage. Thereafter, however, the Rovral@-treated berries decayed at a slightly higher rate than the

chitosan-coatedberries suggestingthe active ingredient of the

fungicide could have been inactivated by the host. Furthermore, Rovral@-treatedberries showed symptoms of phytotox-

14

21

26

35

TIME (days)
Fig. a-Texture
(A) and Titratable acidity (B) of control lb), Rovral@-treated P), l.O%, (0) and 1.5% (0) chitosan-coated strawberries stored at 4C. Vertical bars represent SE values.

icity characterizedby formation of water-soakedareas. At the

end of storage (29 days), decayedberries in control was about
82% and in Rovrala treatment 33%. In contrast, the level of
decay in 1.0 and 1.5% chitosan-coatedberries was 22 and
19%, respectively. There was no added benefit to decay control by increasing concentrationof chitosan from 1.0 to 1.5%.
Chitosan has the capacity to inhibit growth of several fungi,
to induce chitinase, and to elicit phytoalexins in the host tissues. Thus, the control of decay in strawberries could be attributed to either the fungistatic property of chitosan per se or
to its ability to induce defenseenzymes (i.e. chitinase and p1,3-glucanase)and phytoalexins in plants or a combination.
Whatever the mode of action, chitosan proved more effective
than Rovral@in controlling decay of strawberries at 13C.
Effect on quality attributes
Chitosan coating had a beneficial effect on flesh firmness,
titratable acidity and retarding synthesis of anthocyanin of
strawberries stored at 4C (Fig. 2 and 3). Those coated with
chitosan (1.0 and 1.5%), after 31 d were firmer and higher in
titratable acidity than the control or Rovral@-treatedberries
(Fig. 2). Increasing chitosan concentration did not result in
any increasein retention of firmness or modify titratable acidity. Rovral@
treatment was effective in controlling decay, but
did not improve firmness or acidity as comparedto the control.
Chitosan coating delayed rate of ripening as indicated by
anthocyanincontent (Fig. 3). The total anthocyanincontent of
chitosan-coatedberries was the least among the treatments,
after 31 days storage. In addition, the anthocyanin content of
Rovral-treated berries was greater than that of the control.

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CHITOSAN COATING OF STRAWBERRIES. , . .

35 8m

The CO, production of strawberries stored at 4C is presented in Fig. 4. The pattern of respiration for the 1.5% samples was different than those of the control and 1.0% samples.
However, coating with chitosan had an immediate stimulatory
effect on respiration. Such effects gradually disappeared during
the following 2 days. Reduction of respiration rate by chitosan
coating became evident beyond the 4th d of storage. The effect
of chitosan coating on CO, production was greater at higher
concentration.

30 -

c
g

25

5
E
s
.E

20-

95

lo-

lz
E
a

5-

15-

CONCLUSIONS

lb

1;

2b

2;

3b

3;

4'0

4;

Days in storage
Fig. 3-Anthocyanin
content of control @), RovraP-treated
@),
1.0% (0) and 1.5% (e) chitosan-coatedstrawberries
stored at 4C.
Vertical bars represent SE values.

13
I
AZ
.

Effect of chitosan coating on respiration

CHITOSAN COATING was more effective than Rovral@

treatment in controlling postharvest decay of strawberries at
13C. Chitosan-coated berries stored at 4C were firmer and
higher in titratable acidity than Rovral@-treatedor control berries. Chitosan coating neither altered ripening capacity of
strawberries nor caused any apparent phytotoxicity. Our study
indicated that preservative coating with chitosan has a potential
to prolong storage life and control decay of strawberries even
at higher storage temperatures. In addition chitosan coating
with its ability to modify internal atmosphere in the tissue and
fungistatic property can provide a security factor when rigorous
control of storage and distribution temperatures cannot be assured. However, the organoleptic quality of the chitosan coated
fruits remains to be evaluated.

11

REFERENCES
y"
ii
V
c
.o
5
5
t

cu
8
5
0

10

12

14

16

Days in storage
Fig. 4-CO, production of control @), 1.0% (0) and 1.5% (a) chitosan-coated strawberries
stored at 4C. Vertical bars represent
SE values.

This suggested that RovraP treatment may have stimulated the

ripening process. On the contrary, chitosan-coated berries synthesized anthocyanin at a slower rate, and coating neither affected appearance nor caused apparent phytotoxicity.
Furthermore, chitosan-coated berries developed the full red
color after 35 days storage. Retention of firmness, higher titratable acidity and slower rates of anthocyanin production in
coated berries demonstrated that chitosan coating slowed down
metabolism and prolonged the storage life. Coating fruits with
semi-permeable film has generally been shown to retard ripening by modifying the endogenous CO*, O2 and ethylene
levels of fruits (Lowings and Cutts, 1982). However, specific
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Lau, 1988). While available coating materials have been found
to extend storage-life of produce by acting as a diffusion barrier, chitosan coating affords the added advantage of antifungal
activity which is beneficial for highly perishable produce such
as strawberries. In addition chitosan coating is likely to modify
the internal atmosphere without causing anaerobic respiration,
since chitosan films are more selectively permeable to O2 than
to CO, (Bai et al., 1988).
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-Continued
on page 1631