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The aim of this work was to study the effect of A. vera, applied as an edible
coating according to our patent (Martnez-Romero et al., 2003), on the change in physicochemical parameters related to fruit quality during cold storage of pomegranate arils, as
well as its role in controlling microbial spoilage. As far as we are aware, this is the first
published paper in which A. vera gel is used as an edible coating in pomegranate arils. It
could be an innovative and interesting commercial product and an alternative to the use of
synthetic postharvest fungicides.
MATERIALS AND METHODS
Pomegranates (P. granatum L. Mollar de Elche) were picked in a commercial
orchard in Orihuela (Alicante, Spain). Fruit were harvested when fully mature according
to commercial practice and immediately transported to the laboratory. Pomegranates with
defects (sunburn, crack, bruise and cut in the husk) were discarded. The remaining fruit,
around 200 pomegranates, were peeled and washed in order to obtain the arils which were
applied several treatments by immersion as follows. Control arils were immersed in a
solution containing 0.5% citric acid (CA) + 0.5% ascorbic acid (AC). The rest of the fruit
were immersed in a solution containing 50% Aloe vera + 0.5% citric acid (CA) + 0.5%
ascorbic acid (AC). All the arils were packaged in 40 polypropylene trays. All the
packages were stored at 3C during 2 weeks in which internal atmosphere, fruit firmness,
ripening index and microbial counts were studied.
Gas Composition
CO2 and O2 concentrations were quantified in duplicate in each package by
withdrawal of 1 ml of headspace atmosphere using an airtight syringe, and injected into a
gas chromatograph GC 14B (Shimadzu, Tokyo, Japan) equipped with a thermal
conductivity detector (TCD). CO2 and O2 were separated on a molecular sieve 5A
column, 80-100 mesh (Carbosieve SII, Supelco Inc., Bellefonte, USA), of 2 m length and
3 mm i.d. Oven and injector temperature were 50 and 110C, respectively. Helium was
used as carrier gas at a flow rate of 50 ml min-1. Results (meanSE) were expressed as
kPa O2 and kPa CO2 inside the packages.
Fruit Firmness
This parameter was measured using a flat steel plate coupled with a texturometer
(TX-XT2i Texture Analyzer, Stable Microsystems, UK) interfaced to a personal
computer. Arils where place in a compression glass in which was applied a compression
flat steel plate. The force that achieved a 3% deformation of the fruits was applied.
Results were expressed as the force-deformation (N mm-1) and were the mean SE
(n=15).
Ripening Index
Arils of each treatment were combined to obtain a homogenous juice sample for
each replicate, in which total soluble solids (TSS) and total acidity (TA) were determined
in duplicate. TSS concentration was determined with a digital refractometer Atago PR101 (Atago Co. Ltd., Tokyo, Japan) at 20C, and expressed as g 100 g-1. TA (g of malic
acid equivalent per 100 g-1 fresh weight) was determined by automatic titration (785 DMP
Titrino, Metrohm) with 0.1 N NaOH up to pH 8.1, using 1 ml of diluted juice in 25 ml
distilled H2O. The ratio between soluble solids concentration and titratable acidity was
considered as a ripening index (RI).
Microbiological Analysis
Samples of 10 g from each package were obtained under sterile conditions
(laminar fume cupboard, gloves, and scalpels), which were homogenized in 90 ml of
sterile peptone water using a stomacher (model Seward, Laboratory Blender Stomacher
400, London, UK). Serial dilutions were carried out, and 1 ml was added to plate count
agar for mesophilic aerobic and for yeast and mold counts (Petrifilm Aerobic and Yeast
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and Mold Count Plates, Laboratories 3M, Sant, France). Samples were prepared in
triplicate, and only counts of 30-300 colony-forming units (CFU) were considered. Plates
were incubated during 3 days at 30C and 5 days at 25C for mesophilic aerobic and yeast
and mold, respectively.
Statistical Analysis
Data for the physical, chemical, microbiological, and sensory parameters were
subjected to analysis of variance (ANOVA). Sources of variation were time of storage
and treatments. Mean comparisons were performed using HSD of Tukeys test to examine
if differences between treatments and storage time were significant at P<0.05. All
analyses were performed with SPSS software package v. 11.0 for Windows (36).
RESULTS AND DISCUSSION
The CO2 concentration inside the packages in control arils slightly increased over
cold storage, in comparison with Aloe-treated arils. In the same way, O2 concentration
was slightly higher than for control arils in some of the days of sampling but without
significant differences (Fig. 1). On the other hand, ethylene concentration showed a
continuous increase with time in storage for control arils, and this increase was
significantly lower in Aloe treated packages (Fig. 2). The effect of Aloe coating on
reducing these physiological traits could be attributed to modification of the internal
atmosphere (increase in CO2 and decrease in O2) that potentially reduces ethylene
production (Valero and Serrano, 2010). In some non-climacteric fruit, such as table grape,
and sweet cherry, respiration rate was reduced by Aloe in cold storage or in post-storage
shelf conditions at 20C (Valverde et al., 2005; Martnez-Romero et al., 2006).
Texture is an important attribute demanded by consumers and most of the time is
responsible for fruit acceptability. The rate and extension of firmness loss during storage
are the main factors determining fruit quality. In fact, Aloe treatment significantly reduced
the firmness losses during cold storage, whereas losses of >20% were detected in control
arils during cold storage (Fig. 3). The explanation for this firmness maintenance could be
related to the lower weight losses, as has been reported in sweet cherry treated with
different edible coatings (Yaman and Bayindirh, 2002). On the other hand the effect of
A. vera gel on the reduction of -galactosidase, polygalacturonase, and pectinmethylesterase activities, the main cell wall degrading enzymes, cannot be discounted (Nunan et
al., 1998).
The calculation of the ratio between TSS and TA, is considered a ripening index,
and changes showed that there was an advance of the ripening process in control arils
compared with treated ones (Fig. 3). This could be due to the modification of the internal
atmosphere, related to the Aloe coating because the coating produced similar effects as
has been shown for MAP conditions (Serrano et al., 2005). The reduced ripening index
was related to maintenance of TA while the content of TSS was unchanged along storage
irrespective of treatments.
After evaluating total aerobic counts, we observed significant increases in
microbial populations occurred on control fruit, for mesophilic aerobics and yeast and
moulds (Fig. 4). In contrast, microbial populations were significantly reduced in Aloetreated arils, in all samples tested. The antifungal activity of A. vera has been reported
against postharvest fruit pathogens, such as Penicillium digitatum, P. expansum,
B. cinerea and Alternaria alternata (Jasso de Rodrguez et al., 2005) and was based on
the suppression of germination and the inhibition of mycelial growth. In addition, the
inhibitory effects of several Aloe extracts have been also found on Aspergillus niger,
Cladosporium herbarum and Fusarium moniliforme, and could be attributed mainly to the
presence of Aloe-emodin and aloenin together with other active compounds (Ali et al.,
1999), although the specific mechanism of action is still unknown. Moreover, the
reduction of the growth of 17 bacteria by A. vera gel has been proven (Reynolds and
Dweck, 1999), being more effective against gram positive than gram negative
microorganisms (Ferro et al., 2003). Some individual components found in A. vera gel,
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Figures
22
CONTROL
ALOE
5.5
4.5
18
4.0
16
3.5
14
O 2 (%)
CO 2 (%)
CONTROL
ALOE
20
5.0
3.0
2.5
2.0
12
10
8
1.5
1.0
0.5
0.0
0
10
Days of storage at 3C
14
17
10
14
17
Days of storage at 3C
Fig. 1. Changes in CO2 and O2 concentrations (%) of arils inside packages alone (control)
or with Aloe vera coating during cold storage. Data are the meanSE (n=10).
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6.8
CONTROL
ALOE
-1
30
25
20
15
10
CONTROL
ALOE
6.6
6.4
6.2
6.0
5.8
5.6
5.4
5.2
10
Days of storage at 3C
14
17
10
14
17
Days of storage at 3C
Fig. 3. Evolution of fruit firmness (N mm-1) and ripening index (Brix/total acidity) of
pomegranate arils inside packages alone (control) or with Aloe vera coating during
cold storage. Data are the meanSE (n=15).
Fig. 4. Evolution of mesophilic aerobic counts and yeast and molds (CFU ml-1) of
pomegranate arils inside packages alone (control) or with Aloe vera coating during
cold storage. Data are the meanSE (n=10).
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