Microbiological stabilization of Aloe vera (Aloe barbadensis Miller) gel

by high hydrostatic pressure treatments

J.E. Reyes a, G. Tabilo-Munizaga a, M.I. Guanoquiza a, A. Vega-Galvez b, M. Mirandab and M. Prez-Wonb
Food Engineering Department, University of Bio Bio, PO Box 447, Chilln, Chile.
Food Engineering Department, University of la Serena, PO Box 559, La Serena, Chile.
a

ABSTRACT
The effect of high hydrostatic pressure (300, 400 and 500 MPa for 1, 2 and 3 min) on the behaviour of the spoilage microflora and shelf-life extension of Aloe vera gel during 90
days of storage were evaluated. After processing and during storage, there were significant differences in aerobic mesophilic and psychrophilic counts, numbers of
Enterobacteriaceae and fungi, and microbiological shelf-life between untreated (control) and pressurized samples (P<0.05). Immediately after HHP-treatment, counts of
microbial populations studied were reduced to non-detectable levels, except for mesophilic counts in samples treated at 300 MPa for 1 min. The high initial microbial load (>2.0
log CFU ml-1) of the untreated aloe gel limited its shelf-life to 0 days. HHP-treatments at 300 MPa (1, 3 and 5 min) and 400 MPa (1 min) increased the microbiological shelf-life to
18, 24, 35 and 53 days, respectively. Moreover, pressure treatment at or over 400 MPa for 3 min kept the microbial populations below the detection limit during the whole
storage period, leading to a shelf-life over 90 days. This study demonstrated that HPP processing would be an alternative to conventionally thermal processing for the
microbiological stabilization of Aloe vera gel.
INTRODUCTION

RESULTS & DISCUSSION

In recent years, Aloe vera has assumed an

important role in formulation of natural
products both in food and cosmetology
industry. Aloe vera gel has been traditionally
employed
for
having
many
beneficial
properties for human health. Its gel, the
parenchymatic tissue of Aloe leaves, contains
over 98-99% water and more than 60% of the
dry matter is made up of polysaccharides [1].
Due to its therapeutic and
functional
properties and hence its beneficial effects on
humans, the use of Aloe in the formulation
of food products has increased [2]. In view
of its highly perishable nature, mainly from
the microbiology point of view, aloe gel should
be processed in order to extend its shelf-life.
For Aloe vera gel stabilization, it is often
necessary to apply thermal processing, which
can produce irreversible modifications of the
functional
components.
Therefore,
the
challenge to preserve functional properties
suggests the application on non-thermal
innovative technology for the sanitation of
aloe gel such as high hydrostatic pressure
(HHP). HPP is an alternative preservation
process that is proving to be able to inactive
spoilage microoganisms without significantly
affect the natural properties, and so increase
the shelf-life of several foods [3]. Thus, the
aim of this study was to evaluate the
application of HHP on microbial behavior and
the shelf-life extension of the Aloe vera gel
during
cold storage
at 4C.
MATERIALS
& METHODS

The mean initial microbial populations of aerobic mesophilic and psychrophilic microoorganims, Enterobacteriaceae and fungi
in fresh (untreated) Aloe vera gel were 3.76, 3.41, 3.76, 2.57 and 3.36 CFU ml-1 respectively, According to WHO
microbiological standards [5], all counts of microbial populations on untreated samples were higher than the acceptable
limits. Immediately after HHP-treatment, counts of microbial populations studied, except for mesophilic counts in samples
treated at 300 MPa/1 min, were significantly (p< 0.05) reduced to non-detectable levels (<1.0 log CFU ml-1), reaching a
reduction of ca. 2 to 4 log cycle.

10
9
Control
300 MPa - 1 min.
300 MPa - 3 min.
300 MPa - 5 min.
400 MPa - 1 min.
400 MPa - 3 min.
400 MPa - 5 min.
500 MPa - 1 min.
500 MPa - 3 min.
500 MPa - 5 min.

8
7
6
5
4
3
2
1
0
0

10

20

30

40

50

60

70

10
9
Control
300 MPa - 1 min.
300 MPa - 3 min.
300 MPa - 5 min.
400 MPa - 1 min.
400 MPa - 3 min.
400 MPa - 5 min.
500 MPa - 1 min.
500 MPa - 3 min.
500 MPa - 5 min.

8
7
6
5
4
3
2
1
0
0

90

10

20

30

40

50

60

70

80

90

Storage time ( days)

Storage time ( days)

Figure 1. Growth curve of (A) aerobic mesophilic and (B) psychrophilic on aloe gel storage at 4C. Data were fitted by a re-parameterized version of the Gompertz equation.
Bars denote standard deviation of the mean.

During storage at 4C the aerobic mesophilic and psychrophilic counts followed a similar behavior (Figure 1A and 1B). In
control samples, on the first day of storage showed mesophilic counts higher than 2.0 log CFU ml -1, which is considered the
upper acceptable limit for Aloe vera gel by WHO, and were found to be > 8.0 log UFC ml-1 at the end of storage period. In
contrast, in samples treated at 300 MPa (1, 3 and 5 min) and 400 MPa (1 min), this value was reached after 18, 24, 35 and 53
days, respectively. In samples pressurized at or over 400 MPa for 3 min, the counts were kept below the detection limit
during the whole storage period. Enterobacteriaceae and fungi were not detected after any pressure treatment and the
number of survivors was kept below the detection limit during the whole storage (data not shown).

Raw material and gel

preparation.
Selection of
Aloe vera
leaves.

Stabilization
of the gel.
(24h/4C)

Acibar
extraction.

Homogenized
gel.

Cut leaves
of Aloe vera

Homogenization
of the gel
(5min.)

Separation
gel.

Obtaining
the aloe
gel.

The Gompertzs parameters values relative to aerobic mesophilic and psychrophilic

microorganisms for untreated and pressurized aloe gel samples stored a 4C for 90 days are
reported in Table 1. It is evident that the growth kinetic parameters estimated (including the
shelf-life) were significantly (p<0.05) affected for HHP-treatments. HHP-treatments increased
the lag () phase and decreased the growth rate (max) of both microbial populations studied.
Moreover, the microbial shelf-life (SL) increased in pressurized samples. Thus, based on the
aerobic mesophilic counts, the SL estimated was between 18 and 53 days for the samples
treated at 300 MPa/1 min and 400 MPa/1 min, respectively. In contrast, HHP-treatment at or
above 400 MPa for 3 min were able to keep the microbial counts below the detection limit
throughout the cold storage period; therefore, the SL was extended for more than 90 days.
The high initial microbial load (>2.0 log UFC ml-1) found in the untreated aloe gel samples
limited its shelf-life to 0 days.
Treatments
Pressure
(MPa/min)

Treatments.
300 MPa/1, 3 and 5 min.
400 MPa/1, 3 and 5 min.
500 MPa/1, 3 and 5 min.
HHP-treatment.
(Avure Inc., Kent, WA, USA)

Microbiological analysis: Immediately after HHP-treatment and

throughout
subsequent storage at 4C, all samples were analyzed for counts of aerobic
mesophilic and psychrophilic microorganisms, Enterobacteriaceae, and moulds and
yeasts by standard methods.
Estimation of kinetic parameters of growth and shelf-life: The reparameterized Gompertz equation [4] was used to estimate the growth kinetic
parameters, including the shelf-life (SL). The SL was based on aerobic mesophilic
count, taking 2.0 log CFU ml-1 as the upper acceptable limit for Aloe vera gel (WHO,
1999). The equation was fitted to growth data using the nonlinear regression
modulus of the GraphPad Prism v. 4.03 (GraphPad Software, Inc., San Diego, CA,
USA). The goodness of fit was evaluated using the coefficient of determination (R2).

11th INTERNATIONAL CONGRESS ON ENGINEERING AND FOOD

Athens Greece - 2011

Kinetics parameter values

Aeobic mesophilic

Aerobic psychrophilic

max (days-1)

(days)

max (days-1)

0 0.0a

- 0.27 1.22a

0.43 0.05a

- 1.56 1.12a

0.38 0.08a

300/ 1

18 1.9b

18.97 1.55b

0.34 0.04ab

16.98 1.38b

0.33 0.05ab

300/ 3

24 2.0c

19.95 1.85b

0.30 0.03b

17.81 1.72b

0.28 0.02b

300/ 5
400/ 1

35 1.2d
53 1.5e

26.08 2.64c
43.08 1.58d

0.17 0.02c
0.15 0.02c

27.92 2.98c
44.13 2.40d

0.17 0.03c
0.15 0.03c

400/ 3

> 90f

N.De

N.Dd

N.De

N.Dd

400/ 5

> 90f

N.De

N.Dd

N.De

N.Dd

500/ 1

> 90f

N.De

N.Dd

N.De

N.Dd

500/ 3
500/ 5

> 90f
> 90f

N.De
N.De

N.Dd
N.Dd

N.De
N.De

N.Dd
N.Dd

max = The maximum specific growth rate (days -1), = lag phase (days); SL = shelf-life (days). Values are
media of triplicate standard deviation. (a-f) Values in the same column with different superscripts differ
significantly (p < 0.05; LSD). N.D= not determined: parameters were not estimated due to microbial
counts were kept below the detection limit throughout storage.

CONCLUSION
HHP-treatments at or over 400 MPa/3 min were sufficient to reduce the initial
microbial load and extended shelf-life of Aloe vera gel (up to 90 days stored at 4C).
The applications of this technology is particularly interesting on Aloe vera gel,
characterized as high added value, due to its high content of nutraceutical
components, then more studies are needed to determine the effect of HHP-treatment
on the organoleptic and nutraceutical properties.
REFERENCES
1.
2.

Statistical analysis of data: The values of fitting parameters, including the shelflife (SL), were compared for statistical significance using one-way analysis of
variance (ANOVA). Mean separations were obtained using the LSD test. Significance
was determined at the P< 0.05 level. Data processing was carried out using the
statistic software Statgraphics Plus v. 5.1 (Statistical Graphics Co., Rockville, MD,
USA).

SL(days)
(days)

Control (untreated)

High hydrostatic pressure

treatment.

Packaging
Aloe vera gel
in polyethylene
bags.

80

Microbial growth ( log CFU ml-1)

Microbial growth ( logCFU ml-1)

3.
4.
5.

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functional foods: Pomegranate juice. Journal of Food Engineering, 100, 245-253.
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ACKNOWLEDGMENTS
This study was financially supported by FONDECYT program (project 1090228).