Introduction
As a result of the increase in the human population, agriculture techniques must be
improved to face the challenge of feeding humanity with healthy and nutritious food,
exchanging ancient methods for the use of chemical substances that enhance growth
and reduce losses due to pests (Hahn, 2014).
Moreover, the trend of eating fresh products for an appropriated diet has
consequently led to food being processed faster, which increases the risk of
contamination by pesticides, toxins, or microorganisms (Jung, Jang & Matthews, 2014).
In the context of agricultural production, there are risks associated with the loss
of crops at all stages of the process. However, post-harvest losses are one of the
most significant since they occur immediately prior to the product reaching the
consumer (Kasso & Bekele, 2018).
Post-harvest activities represent 34% of global crop losses, with the most frequent
causes being pests, natural ripening processes, environmental conditions and
microbial infection mainly with fungi (Abass et al.,
2014; Kasso & Bekele, 2018;
Kumar & Kalita, 2017).
The infection of harvested crops with phytopathogenic fungi represents one of the
most important challenges in current agricultural techniques, not only from an
economic point of view but for the risk to human health. Several factors are related
to fungal infection under post-harvest conditions, such as pH, temperature, UV and
oxidative stress, among others (Liu et al.,
2018). In recent decades, the use of organic fungicides (such as
site-specific fungicides, thiabendazole, pyrimethanyl, o-phenylphenol and imazalil),
has been the main force in avoiding crop decay due to their effectiveness and
spectrum of action. However, the use of these substances has a serious environmental
impact and causes human health disorders and antimicrobial resistance (Hahn, 2014; Peris-Vicente, Marzo-Mas, Roca-Genovés, Carda-Broch & Esteve-Romero,
2016).
The current trends in crop protection involve a range of actions that mitigate the
environmental impact, have a broad spectrum of action and are of natural origin,
biodegradable and easy to apply (Seiber, Coats, Duke
& Gross, 2014). One alternative is the use of natural materials such
as chitosan and Opuntia mucilage as edible film presentation.
Chitosan (CS) is one of the most promising resources for the generation of bioactive,
biodegradable materials against infections that affect crops of economic importance.
The main chain of this semi-natural polymer displays a polycationic behavior in
acidic solution due to the free amino groups in its structure (Olicón-Hernández, Uribe-Álvarez, Uribe-Carvajal, Pardo &
Guerra-Sánchez, 2017). This polymer is obtained by the deacetylation of
chitin, the second most abundant structural polymer related to fungi, crustaceans
and insects (Olicón-Hernández, Zepeda Giraud &
Guerra-Sánchez, 2017). Due to its properties, such as resistance,
flexibility, polymerization, and polycationic charge, chitosan has been used as a
raw material for the formation of gels, fibers and films and has been shown to have
extensive antimicrobial activity against bacteria and fungi (Anitha et al., 2014; Olicón-Hernández et al., 2015; Olicón-Hernández, Zepeda Giraud, et al., 2017;
Younes & Rinaudo, 2015).
In the context of the edible films formulation, a typical edible film has three major
components; film forming material, plasticizer, and additives. From this point of
view Nopal (Opuntia ficus-indica) is a potential alternative as
element of edible film. The nopal is a traditional element of mexican food and
culture but is considered an exotic ingredient in an international context (de Albuquerque et al., 2018). Extracts of this
plant have shown benefits to human health, such as hypolipidemic,
hypocholesterolemic, antidiabetic, hypoglycemic, antioxidantand anti-inflammatory
efficacies (Otálora, Carriazo, Iturriaga, Nazareno
& Osorio, 2015). Opuntia mucilage (OM) is a complex
material formed from carbohydrates including L-arabinose, D-galactose, D-xylose,
L-rhamnose and D-galacturonic acid in variable proportions, which has been cataloged
as a promising natural resource for use in the food, pharmaceutical and construction
industries (León-Martínez, Cano-Barrita,
Lagunez-Rivera & Medina-Torres, 2014; Otálora et al., 2015).
In the present study, we designed a CS-OM film for the protection of crops, which has
the characteristic of being edible, allowing safe and practical handling for its
application. Physicochemical characterization of the film was performed and the
in vitro and in situ antifungal activity
against Rhizopus stolonifer, one of the most important
phytopathogenic fungi, was evaluated.
Materials and Methods
Regents and Solutions
Low-molecular weight chitosan (50,000−190,000 Da) was provided by Sigma−Aldrich
(St. Louis, MO, USA). A stock solution of 10 mg/mL was made in 1% acetic acid,
according to our previously reported protocol, followed by sterilization via
autoclaving (Olicón-Hernández et al.,
2015).
The Opuntia (nopal) cladodes for OM extraction were obtained
from a local market in México City. The cladodes were thoroughly washed, cut
into uniform pieces (0.5 cm x 0.5 cm), placed in water (5:1 w/v) and incubated
overnight at room temperature. Subsequently, the mixture was filtered and
concentrated by evaporation.
Glycerol and gelatin were of commercial food grade and purchased from a local
supermarket.
CS-OM film formulation
The film formulation was made by mixing four main elements: 1) CS, 2) OM, 3)
glycerol and 4) gelatin. The proportions of OM and gelatin kept constant (20 and
10%, respectively) to maintain the integrity of the membrane. Eight different
preparations were made, with two concentrations of glycerol (1 and 3%), to
evaluated the flexibility and four concentrations of chitosan (0.05, 0.1, 0.15
and 0.20%), to test the homogeneity and antifungal effect of the resulting film
(Table I). Each mixture was homogenized using a vortex and placed into Petri
dishes (8.7 cm), at room temperature until dry. The film was manually separated
from the plate. Selection was made based on the integrity, flexibility and
handling of the resulting film, in addition to the homogeneity, as confirmed by
conventional microscopy.
Scanning electron microscopy (SEM)
The selected film was cut into 1cm x 1cm pieces and placed on metallic bases for
structural evaluation by SEM. After the samples were dried, a gold layer was
directly applied under a vacuum for 10 min and observed using a JEOL 5800LV
scanning electron microscope (Tokyo, Japan) at 15 kV (Olicón-Hernández et al., 2015).
Physicochemical characterization
Viscosity
The viscosity of the selected formulation was determined as a characterization
element. Likewise, the viscosity of each of the elements of the mixture was
determined separately to evaluate its rheological behavior. A viscometer
Brookfield model RVT Spring Torque (Dyne-cm), 7,187.0 (Middleboro, MA, USA), was
used. Six spindles and eight velocities were tested to standardize the
determination of each component (manual instructions). A volume of 600 mL sample
was placed in a beaker and the spindle of the viscometer was submerged and
centered in the sample. Rotation of the viscometer was maintained until the
reading stabilized and the determination of viscosity was carried out according
to the specifications described in the manual (Viscometer, 2016). The experiments were performed in triplicate and
the viscosity is expressed in centipoises (cp).
Color
The color determination of the film was carried out using a CR-10 colorimeter
(Konica Minolta, Japan). The determination was randomly performed on the surface
of three different samples and the L*, a*and b* values were measured based on
the CIE Lab* color space (Valadez-Carmona et
al., 2016).
Percentage humidity and film solubility in water
The humidity was measured by gravimetric standard methods and is expressed as a
percentage (%). The experiments were performed in triplicate. Percentage
solubility in water was measured according to García et al.,
2004, based on the comparison of the weight of the dry sample prior to and
following exhaustive agitation in water, as calculated using the following
equation (Garcı́a, Pinotti, Martino &
Zaritzky, 2004):
%Solubility=Initial dry weight-Final dry weightInitial dry weight*100
Qualitative evaluation of the protection of stored fruits
Ten Saladette tomatoes (Lycopersicon esculentum) were selected
according to color, state of health, size and firmness. These fruits were
divided into two equal groups: one as a control without protection and the other
protected by adhesion of the CS-OM on the surface. The two groups were stored at
room temperature and humidity for 30 days. No phytopathogenic agent was
intentionally added. After this time, a visual evaluation was made to determine
the protective effect of the film as compared with the control.
Antifungal activity
Rhizopus stolonifer R3 was isolated from post-harvest
contaminated crops and also provided by CEPROBI-IPN, Yautepec, Morelos, Mexico
(Hernández-Lauzardo et al.,
2008).
Determination of in vitro antifungal
activity
Rhizopus stolonifer R3 was grown on commercial potato dextrose
agar (PDA) plates for 72 h. After this time, approximately 1-cm2
pieces of mycelia-agar were excised to inoculate three PDA plates without CS-OM
film (control system, Db) and three plates coated with the film (positive
system, Da). The plates were incubated at 28 °C until the fungus in the control
system covered the plate. In both systems, the diameter of the mycelia was
measured to determine the antifungal index according to the following equation
(Guo et al., 2006):
Antifungal Index%=1-DaDb*100
where Da is the diameter of the mycelia in the positive system and Db is the
diameter in the control system.
Determination of in situ antifungal
activity
R. stolonifer R3 was grown on PDA plates for 72 h at 28 °C.
After this time, a spore suspension was made by mechanical extraction in water.
The number of spores was calculated using the Neubauer chamber method and
adjusted to 1x106 spores/mL in sterile water (Olicón-Hernández, Camacho-Morales, Pozo, González-López &
Aranda, 2019).
A total of 90 Saladette tomatoes (Lycopersicon esculentum) were
selected using the same parameters as described in section 2.5. These tomatoes
were extensively washed, disinfected with 1% sodium hypochlorite and divided
into six groups of 15 samples. Three of these groups were protected with the
CS-OM film (test groups) and three groups were the control. The spore solution
was sprinkled on all tomatoes, which were subsequently placed in humid chambers
at room temperature for 72 h. After this time, the severity index (damage) was
calculated according to the protocol-scale-equation described by Mayek−Perez et al., 1995, based on the
ratio of samples with visible symptoms of infection to the total samples in the
group (Mayek-Pérez, Pedroza-Flores,
Villarreal-García & Valdés-Lozano, 1995).
Results
Selection of the CS-OM film
Evaluation of the different formulations is displayed in Table I. In general, a low proportion of glycerol resulted
in lumpy films with a lack of continuity, which did not allow complete drying.
On the other hand, micrographs of the films (Figure 1) revealed that by increasing the concentration of chitosan,
the surface was more homogeneous and the flexibility was improved. According to
these results, the formulation containing 3% glycerol and 2% chitosan was
selected, with the proportions of gelatin and mucilage as specified in section
2.2.
Table I
Evaluation of the physical characteristics of the different
formulations of CS-OM film.
|
Formulation |
Chitosan (%) |
Glycerol (%) |
Integrity |
Flexibility |
Homogeneity |
|
1 |
0.05 |
1.0 |
+ |
+ |
+ |
|
2 |
0.10 |
|
+ |
++ |
+ |
|
3 |
0.15 |
|
++ |
++ |
+ |
|
4 |
0.20 |
|
+++ |
+++ |
+ |
|
5 |
0.05 |
3.0 |
++ |
++ |
++ |
|
6 |
0.10 |
|
++ |
++ |
+++ |
|
7 |
0.15 |
|
+++ |
++ |
++++ |
|
8 |
0.20 |
|
++++ |
++++ |
++++ |
Figure 1
Micrographs of the CS-OM film formulations (100x). Evaluation of
the properties of the film was carried out qualitatively. (A) 1%
glycerol; (B) 3% glycerol. 1 and 5 = 0.5% chitosan; 2 and 6 = 1%
chitosan; 3 and 7 = 1.5% chitosan; 4 and 8 = 2% chitosan.
Characterization of the CS-OM film
According to SEM analysis, the CS-OM film had a fibrous structure with the
absence of pores and a homogenous appearance throughout its surface (Figure 2), which is in accordance with the
simple light microscopy data.
Figure 2
SEM analysis of the CS-OM film. No damage or irregularity was
observed in the structure of the film following the drying process.
The fibrous structure originated from the interaction of the
different components, with chitosan conferring that property. (A)
500x; (B) 1500x; (C) 4000x.
The data obtained from the individual elements and the aqueous phase of the CS-OM
film (Figure 3), demonstrated that the
viscosity was cumulative, with glycerol as the most important element affecting
this property.
Figure 3
Viscosity determination of the CS-OM film elements. The viscosity
is expressed in centipoises. OM = Opuntia mucilage; CS = Chitosan;
CS-OM film = aqueous phase of the designed film.
The data corresponding to humidity, solubility and color are summarized in Table II. Following drying, the resulting
film had an average of 12.5% and 43% humidity and solubility, respectively.
According to the scales of the L*, a* and b* parameters, the CS-OM film was
slightly dark with a light green-blue coloration.
Table II
Percentages of humidity and solubility and color parameters of
CS-OM films.
|
Humidity (%) |
Solubility (%) |
Color |
|
White calibration |
Black calibration |
|
L* |
a* |
b* |
L* |
a* |
b* |
|
12.4 ± 0.1 |
43.6 ± 0.1 |
43.8 ± 3.81 |
-3.1 ± 0.42 |
9.0 ± 1.97 |
32.9 ± 0.84 |
-1.45 ± 0.35 |
4.05 ± 0.07 |
Effect on the protection of stored fruits
Changes in tomatoes with and without (control) CS-OM film after 30 days of
storage are shown in Figure 4. In the
control, it was observed that the fruits without protection lost firmness and
had slight changes in color, in addition to there being clear evidence of fungal
disease. It should be noted that for this determination, no phytopathogenic
agent was intentionally added. In contrast, the protected fruits maintained
uniform coloration and firmness comparable to the tomatoes under initial storage
conditions and no symptoms of the microbial disease were observed.
Figure 4
Changes in stored fruits with and without CS-OM film. (A,
Control) Fruits without CS-OM film (B) and fruits protected by CS-OM
film. The fruits were stored for 30 days. In control fruits, changes
in color, texture and firmness were observed and fungal mycelia were
present on the surface.
In vitro and in situ antifungal
activity
Changes in the growth of R. stolonifer with and without CS-OM
film on the PDA plates and determination of the antifungal index (%) are shown
in Figure 5. The presence of the CS-OM film
inhibited the growth of the phytopathogenic agent by approximately 50%. This
difference was statistically validated using Tukey’s test (P
< 0.001).
Figure 5
In vitro antifungal effect of CS-OM film against
R. stolonifer. Mycelial growth without film (A,
Control) and with the protection of film (B). The fungus took 36
hours to invade the control plates. There was a statistically
significant difference validated by Tukey’s test (P <
0.001).
In the case of in situ antifungal activity, the tomatoes in the
control group (Figure 6A), showed an
abundant growth of fungus, which in some cases covered more than 50% of the
total surface. During this stage of infection, symptoms of the disease or
important changes in coloration or form were not appreciated. However, the
severity index in this group reached the maximum value on the scale, indicating
that more than 75% of the total fruits were affected.
Figure 6
In situ antifungal effect of CS-OM film against
R. stolonifer. Fruits without film (A,
Control), had massive mycelial growth on the surface. On the
tomatoes protected by CS-OM film (B), a lower and limited fungal
growth was observed.
On the other hand, the fruits protected by the CS-OM film (Figure 6B), obtained a severity index of 2, indicating that
less than 25% of the fruits were affected to some degree. Growth in the area of
affected fruits was lower and limited to specific areas.
Discussion
The use of edible films for the protection of crops, fruits and/or meat is a current
trend with a view to increasing shelf-life, maximizing productivity and maintaining
the quality and flavors of food (Zambrano-Zaragoza
et al., 2018). In general, the composition of edible biofilms requires a
protein component, a plasticizing agent and in some cases a structural component
and/or bioactive agent (Galus & Kadzińska,
2015). In the present work, gelatin and glycerol were selected as the
protein and plasticizing agent, respectively, since it has been reported that the
combination of these components improves the quality of film and increases the
shelf-life of fruits such as avocado and strawberries (Aguilar-Méndez, Martín-Martínez, Tomás, Cruz-Orea, & Jaime-Fonseca,
2008; Del-Valle, Hernández-Muñoz, Guarda
& Galotto, 2005).
Several components have been used as elements in film formulation to improve the
flexibility and integrity of the coating. Casein, collagen, lipids, soy, corn,
alginate, cellulose and essential oils are substances that have been proven to be
efficient structural, gelling and active agents (Galus & Kadzińska, 2015). In the present work, the use of
Opuntia mucilage, an unconventional cheap raw material that
contributes carbohydrates, as well as structural and functional elements that make
the film edible and complement its biological activity, were explored. In this
context, the use of Opuntia mucilage as an element in edible films
increases water vapor permeability (Domínguez-Martínez et al., 2017), maintains the quality
of fruits and vegetables (Allegra et al.,
2016; Del-Valle et al., 2005; Treviño-Garza, García, Heredia, Alanís-Guzmán &
Arévalo-Niño, 2017)and improves thermal stability and flexibility (Guadarrama-Lezama et al., 2018). Moreover, the
combination of Opuntia mucilage and chitosan as a bioactive
material improves the structure, stability and homogeneity of the film, as observed
by SEM analysis and fundamental parameters (Maqbool,
Ali, Alderson, Zahid & Siddiqui, 2011). The physicochemical and
rheological characteristics of the CS-OM film in the present study are in accordance
with those previously reported, where there is no standard trend in these parameters
and they depend on the composition and proportions in each particular case (Razavi, Mohammad Amini & Zahedi, 2015;
Saberi & Golding, 2018). However,
maintaining the stability and the physicochemical characteristics of the film is
important prospective work, since these can modify the initial sensory
characteristics of the product and affect the perception of potential buyers
The use of chitosan as an antifungal element is a widely explored field. However, the
underlying mechanisms have not yet been fully elucidated. We propose a global
mechanism that includes electrostatic interactions with the plasma membrane (the
most accepted model) and a series of stress conditioning elements (such as the
formation of reactive oxygen species) and a complex metabolic imbalance (Olicón-Hernández et al., 2015; Olicón-Hernández, Uribe-Álvarez et
al., 2017). With respect to the effect of chitosan on
R. stolonifer, it was observed that the polymer modified the
germination of spores, affected fungal morphology, increased glucose consumption,
inhibited H+-ATPase activity in the plasma membrane and caused a high
potassium efflux (Dos Santos et al., 2012;
García-Rincón et al., 2010; Guerra-Sánchez, Vega-Pérez, Velázquez-del Valle &
Hernández-Lauzardo, 2009; Hernández-Lauzardo et al., 2008; Hernández-Lauzardo et al., 2011). An increase
in the shelf-life of different fruits using edible CS-OM films has been observed in
recent years, which is in agreement with the results of the present study (Ávila-Sosa et al., 2012; Treviño-Garza et al., 2017). However, in most of the cases, the
films were mixed with essential oils or adjuvants to increase the antifungal effect
(Yuan, Chen & Li, 2016) and few
recent studies evaluate the effectiveness of these films under in
situ or in vitro conditions.
As the results indicate, the presence of the polymer mixed with the
Opuntia mucilage inhibited the proliferation of the fungus
under in vitro and in situ conditions for the
protection of tomatoes. These results are consistent with those reported by
Martínez-Camacho et al., who determined similar antifungal index
values against Aspergillus niger. Nevertheless, in
situ protection was not tested by them (Martínez-Camacho et al., 2010). Recently, Robledo et al., 2018, described thymol
nanoemulsions incorporated into quinoa protein/chitosan edible films for the
protection of cherry tomatoes, which significantly reduced fungal infection by
Botrytis cinerea following incubation at 5 °C for seven days
(Robledo et al., 2018).
In summary, the results obtained in the present study describe an attractive
potential alternative that can be used under real conditions for the protection of
fruits against infection with phytopathogenic fungi.
Conclusion
An edible film was designed based on chitosan and Opuntia mucilage
that presented a uniform and homogenous surface/structure. The best integrity was
observed in the film containing the highest proportions of glycerol and chitosan,
which affected its stability and antifungal effect. The CS-OM film significantly
reduced the growth of R. stolonifer under in vitro
and in situ conditions for the protection of tomatoes and increased
their shelf-life, making it a potential product for possible massive application in
agriculture.
Conflict of Interest Statement
The authors declare that they have no conflict of interest.
Acknowledgments
This paper was supported by CONACYT grant 256520 and SIP-IPN (Secretaría de
Investigación y Posgrado Instituto Politécnico Nacional) grant 20190200. The main
author is a postdoctoral fellow of the Dirección General de Asuntos del Personal
Académico (DGAPA) program of Universidad Nacional Autónoma de México.
References
Abass, A. B., Ndunguru, G., Mamiro, P., Alenkhe, B., Mlingi, N.
& Bekunda, M. (2014). Post-harvest food losses in a maize-based farming
system of semi-arid savannah area of Tanzania. Journal of Stored
Products Research, 57, 49-57.
DOI:https://doi.org/10.1016/j.jspr.2013.12.004.
A. B. Abass
G. Ndunguru
P. Mamiro
B. Alenkhe
N. Mlingi
M. Bekunda
2014Post-harvest food losses in a maize-based farming system of
semi-arid savannah area of TanzaniaJournal of Stored Products Research57495710.1016/j.jspr.2013.12.004
Aguilar-Méndez, M. A., Martín-Martínez, E. S., Tomás, S. A.,
Cruz-Orea, A. & Jaime-Fonseca, M. R. (2008). Gelatine-starch films:
Physicochemical properties and their application in extending the post-harvest
shelf life of avocado (Persea americana). Journal of
the Science of Food and Agriculture, 88(2), 185-193.
DOI:10.1002/jsfa.3068.
M. A. Aguilar-Méndez
E. S. Martín-Martínez
S. A. Tomás
A. Cruz-Orea
M. R. Jaime-Fonseca
2008Gelatine-starch films: Physicochemical properties and their
application in extending the post-harvest shelf life of avocado (Persea
americana)Journal of the Science of Food and Agriculture88(2)18519310.1002/jsfa.3068
Allegra, A., Inglese, P., Sortino, G., Settanni, L., Todaro, A.
& Liguori, G. (2016). The influence of Opuntia ficus-indica
mucilage edible coating on the quality of ‘Hayward’ kiwifruit slices.
Postharvest Biology and Technology, 120, 45-51.
DOI:https://doi.org/10.1016/j.postharvbio.2016.05.011.
A. Allegra
P. Inglese
G. Sortino
L. Settanni
A. Todaro
G. Liguori
2016The influence of Opuntia ficus-indica mucilage edible coating on
the quality of ‘Hayward’ kiwifruit slicesPostharvest Biology and Technology120455110.1016/j.postharvbio.2016.05.011
Anitha, A., Sowmya, S., Kumar, P. T. S., Deepthi, S., Chennazhi, K.
P., Ehrlich, H., Tsurkan, M. & Jayakumar, R. (2014). Chitin and chitosan in
selected biomedical applications. Progress in Polymer Science,
39(9), 1644-1667.
DOI:https://doi.org/10.1016/j.progpolymsci.2014.02.008.
A. Anitha
S. Sowmya
P. T. S. Kumar
S. Deepthi
K. P. Chennazhi
H. Ehrlich
M. Tsurkan
R. Jayakumar
2014Chitin and chitosan in selected biomedical
applicationsProgress in Polymer Science39(9)1644166710.1016/j.progpolymsci.2014.02.008
Ávila-Sosa, R., Palou, E., Jiménez Munguía, M. T.,
Nevárez-Moorillón, G. V., Navarro Cruz, A. R. & López-Malo, A. (2012).
Antifungal activity by vapor contact of essential oils added to amaranth,
chitosan, or starch edible films. International Journal of Food
Microbiology, 153(1), 66-72.
DOI:https://doi.org/10.1016/j.ijfoodmicro.2011.10.017.
R. Ávila-Sosa
E. Palou
M. T. Jiménez Munguía
G. V. Nevárez-Moorillón
A. R. Navarro Cruz
A. López-Malo
2012Antifungal activity by vapor contact of essential oils added to
amaranth, chitosan, or starch edible filmsInternational Journal of Food Microbiology153(1)667210.1016/j.ijfoodmicro.2011.10.017
de Albuquerque, J. G., de Souza Aquino, J., de Albuquerque, J. G.,
de Farias, T. G. S., Escalona-Buendía, H. B., Bosquez-Molina, E. & Azoubel,
P. M. (2018). Consumer perception and use of nopal (Opuntia
ficus-indica): A cross-cultural study between Mexico and Brazil.
Food Research International, 124(1),101-108.
DOI:https://doi.org/10.1016/j.foodres.2018.08.036.
J. G. de Albuquerque
J. de Souza Aquino
J. G. de Albuquerque
T. G. S. de Farias
H. B. Escalona-Buendía
E. Bosquez-Molina
P. M. Azoubel
2018Consumer perception and use of nopal (Opuntia ficus-indica): A
cross-cultural study between Mexico and BrazilFood Research International124(1),10110810.1016/j.foodres.2018.08.036
Del-Valle, V., Hernández-Muñoz, P., Guarda, A. & Galotto, M. J.
(2005). Development of a cactus-mucilage edible coating (Opuntia ficus
indica) and its application to extend strawberry (Fragaria
ananassa) shelf-life. Food Chemistry, 91(4),
751-756. DOI:https://doi.org/10.1016/j.foodchem.2004.07.002.
V. Del-Valle
P. Hernández-Muñoz
A. Guarda
M. J. Galotto
2005Development of a cactus-mucilage edible coating (Opuntia ficus
indica) and its application to extend strawberry (Fragaria ananassa)
shelf-lifeFood Chemistry91(4)75175610.1016/j.foodchem.2004.07.002
Domínguez-Martinez, B. M., Martínez-Flores, H. E., Berrios, J. D.
J., Otoni, C. G., Wood, D. F. & Velázquez, G. (2017). Physical
Characterization of Biodegradable Films Based on Chitosan, Polyvinyl Alcohol and
Opuntia mucilage. Journal of Polymers and the
Environment, 25(3), 683-691.
DOI:10.1007/s10924-016-0851-y.
B. M. Domínguez-Martinez
H. E. Martínez-Flores
J. D. J. Berrios
C. G. Otoni
D. F. Wood
G. Velázquez
2017Physical Characterization of Biodegradable Films Based on
Chitosan, Polyvinyl Alcohol and Opuntia mucilageJournal of Polymers and the Environment25(3)68369110.1007/s10924-016-0851-y
Dos Santos, N. S. T., Athayde Aguiar, A. J. A., de Oliveira, C. E.
V., Veríssimo de Sales, C., de Melo e Silva, S., Sousa da Silva, R., Montenegro
Stamford, T.C. & de Souza, E. L. (2012). Efficacy of the application of a
coating composed of chitosan and Origanum vulgare L. essential
oil to control Rhizopus stolonifer and Aspergillus
niger in grapes (Vitis labrusca L.). Food
Microbiology, 32(2), 345-353.
DOI:https://doi.org/10.1016/j.fm.2012.07.014.
N. S. T. Dos Santos
A. J. A. Athayde Aguiar
C. E. V. de Oliveira
C. Veríssimo de Sales
S. de Melo e Silva
R. Sousa da Silva
T.C. Montenegro Stamford
E. L. de Souza
2012Efficacy of the application of a coating composed of chitosan and
Origanum vulgare L. essential oil to control Rhizopus stolonifer and
Aspergillus niger in grapes (Vitis labrusca L.)Food Microbiology32(2)34535310.1016/j.fm.2012.07.014
Galus, S. & Kadzińska, J. (2015). Food applications of
emulsion-based edible films and coatings. Trends in Food Science &
Technology, 45(2), 273-283.
DOI:https://doi.org/10.1016/j.tifs.2015.07.011.
S. Galus
J. Kadzińska
2015Food applications of emulsion-based edible films and
coatingsTrends in Food Science & Technology45(2)27328310.1016/j.tifs.2015.07.011
Garcı́a, M. a. A., Pinotti, A., Martino, M. N. & Zaritzky, N. E.
(2004). Characterization of composite hydrocolloid films. Carbohydrate
Polymers, 56(3), 339-345.
DOI:https://doi.org/10.1016/j.carbpol.2004.03.003.
M. a. A. Garcı́a
A. Pinotti
M. N. Martino
N. E. Zaritzky
2004Characterization of composite hydrocolloid filmsCarbohydrate Polymers56(3)33934510.1016/j.carbpol.2004.03.003
García-Rincón, J., Vega-Pérez, J., Guerra-Sánchez, M. G.,
Hernández-Lauzardo, A. N., Peña-Díaz, A. & Velázquez-Del Valle, M. G.
(2010). Effect of chitosan on growth and plasma membrane properties of
Rhizopus stolonifer (Ehrenb.:Fr.) Vuill. Pesticide
Biochemistry and Physiology, 97(3), 275-278.
DOI:https://doi.org/10.1016/j.pestbp.2010.03.008.
J. García-Rincón
J. Vega-Pérez
M. G. Guerra-Sánchez
A. N. Hernández-Lauzardo
A. Peña-Díaz
M. G. Velázquez-Del Valle
2010Effect of chitosan on growth and plasma membrane properties of
Rhizopus stolonifer (Ehrenb.:Fr.) VuillPesticide Biochemistry and Physiology97(3)27527810.1016/j.pestbp.2010.03.008
Guadarrama-Lezama, A. Y., Castaño, J., Velázquez, G.,
Carrillo-Navas, H. & Álvarez-Ramírez, J. (2018). Effect of nopal mucilage
addition on physical, barrier and mechanical properties of citric pectin-based
films. Journal of Food Science and Technology, 55(9), 3739-3748.
DOI:10.1007/s13197-018-3304-x.
A. Y. Guadarrama-Lezama
J. Castaño
G. Velázquez
H. Carrillo-Navas
J. Álvarez-Ramírez
2018Effect of nopal mucilage addition on physical, barrier and
mechanical properties of citric pectin-based filmsJournal of Food Science and Technology55(9)3739374810.1007/s13197-018-3304-x
Guerra-Sánchez, M. G., Vega-Pérez, J., Velázquez-del Valle, M. G.,
& Hernández-Lauzardo, A. N. (2009). Antifungal activity and release of
compounds on Rhizopus stolonifer (Ehrenb.:Fr.) Vuill. by effect
of chitosan with different molecular weights. Pesticide Biochemistry and
Physiology, 93(1), 18-22.
DOI:https://doi.org/10.1016/j.pestbp.2008.09.001.
M. G. Guerra-Sánchez
J. Vega-Pérez
M. G. Velázquez-del Valle
A. N. Hernández-Lauzardo
2009Antifungal activity and release of compounds on Rhizopus
stolonifer (Ehrenb.:Fr.) Vuill. by effect of chitosan with different
molecular weightsPesticide Biochemistry and Physiology93(1)182210.1016/j.pestbp.2008.09.001
Guo, Z., Chen, R., Xing, R., Liu, S., Yu, H., Wang, P., Li, C. &
Li, P. (2006). Novel derivatives of chitosan and their antifungal activities
in vitro. Carbohydrate Research, 341(3),
351-354. DOI:https://doi.org/10.1016/j.carres.2005.11.002.
Z. Guo
R. Chen
R. Xing
S. Liu
H. Yu
P. Wang
C. Li
P. Li
2006Novel derivatives of chitosan and their antifungal activities in
vitroCarbohydrate Research341(3)35135410.1016/j.carres.2005.11.002
Hahn, M. (2014). The rising threat of fungicide resistance in plant
pathogenic fungi: Botrytis as a case study. Journal of
Chemical Biology, 7(4), 133-141.
DOI:10.1007/s12154-014-0113-1.
M. Hahn
2014The rising threat of fungicide resistance in plant pathogenic
fungi: Botrytis as a case studyJournal of Chemical Biology7(4)13314110.1007/s12154-014-0113-1
Hernández-Lauzardo, A. N., Bautista-Baños, S., Velázquez-del Valle,
M. G., Méndez-Montealvo, M. G., Sánchez-Rivera, M. M. & Bello-Pérez, L. A.
(2008). Antifungal effects of chitosan with different molecular weights on
in vitro development of Rhizopus
stolonifer (Ehrenb.:Fr.) Vuill. Carbohydrate
Polymers, 73(4),
541-547.DOI:https://doi.org/10.1016/j.carbpol.2007.12.020.
A. N. Hernández-Lauzardo
S. Bautista-Baños
M. G. Velázquez-del Valle
M. G. Méndez-Montealvo
M. M. Sánchez-Rivera
L. A. Bello-Pérez
2008Antifungal effects of chitosan with different molecular weights
on in vitro development of Rhizopus stolonifer (Ehrenb.:Fr.)
VuillCarbohydrate Polymers73(4)54154710.1016/j.carbpol.2007.12.020
Hernández-Lauzardo, A. N., Vega-Pérez, J., Velázquez-del Valle, M.
G., Sánchez, N. S., Peña, A. & Guerra-Sánchez, G. (2011). Changes in the
Functionality of Plasma Membrane of Rhizopus stolonifer by
Addition of Chitosan. Journal of Phytopathology, 159(7-8),
563-568. DOI:10.1111/j.1439-0434.2011.01802.x.
A. N. Hernández-Lauzardo
J. Vega-Pérez
M. G. Velázquez-del Valle
N. S. Sánchez
A. Peña
G. Guerra-Sánchez
2011Changes in the Functionality of Plasma Membrane of Rhizopus
stolonifer by Addition of ChitosanJournal of Phytopathology159(7-8)56356810.1111/j.1439-0434.2011.01802.x
Jung, Y., Jang, H. & Matthews, K. R. (2014). Effect of the food
production chain from farm practices to vegetable processing on outbreak
incidence. Microbial Biotechnology, 7(6), 517-527.
DOI:10.1111/1751-7915.12178.
Y. Jung
H. Jang
K. R. Matthews
2014Effect of the food production chain from farm practices to
vegetable processing on outbreak incidenceMicrobial Biotechnology7(6)51752710.1111/1751-7915.12178
Kasso, M. & Bekele, A. (2018). Post-harvest loss and quality
deterioration of horticultural crops in Dire Dawa Region, Ethiopia.
Journal of the Saudi Society of Agricultural Sciences,
17(1), 88-96. DOI:https://doi.org/10.1016/j.jssas.2016.01.005.
M. Kasso
A. Bekele
2018Post-harvest loss and quality deterioration of horticultural
crops in Dire Dawa Region, EthiopiaJournal of the Saudi Society of Agricultural Sciences17(1)889610.1016/j.jssas.2016.01.005
Kumar, D. & Kalita, P. (2017). Reducing Postharvest Losses
during Storage of Grain Crops to Strengthen Food Security in Developing
Countries. Foods MDPI, 6(1), 8.
D. Kumar
P. Kalita
2017Reducing Postharvest Losses during Storage of Grain Crops to
Strengthen Food Security in Developing CountriesFoods MDPI6(1)88
León-Martínez, F. M., Cano-Barrita, P. F. d. J., Lagunez-Rivera, L.,
& Medina-Torres, L. (2014). Study of nopal mucilage and marine brown algae
extract as viscosity-enhancing admixtures for cement based materials.
Construction and Building Materials, 53, 190-202.
DOI:https://doi.org/10.1016/j.conbuildmat.2013.11.068.
F. M. León-Martínez
P. F. d. J. Cano-Barrita
L. Lagunez-Rivera
L. Medina-Torres
2014Study of nopal mucilage and marine brown algae extract as
viscosity-enhancing admixtures for cement based materialsConstruction and Building Materials53190202:10.1016/j.conbuildmat.2013.11.068
Liu, J., Sui, Y., Wisniewski, M., Xie, Z., Liu, Y., You, Y.,Zhang,
X., Sun, Z., Li, W., Li, Y. & Wang, Q. (2018). The impact of the postharvest
environment on the viability and virulence of decay fungi. Critical
Reviews in Food Science and Nutrition, 58(10), 1681-1687.
DOI:10.1080/10408398.2017.1279122.
J. Liu
Y. Sui
M. Wisniewski
Z. Xie
Y. Liu
Y. You
X. Zhang
Z. Sun
W. Li
Y. Li
Q. Wang
2018The impact of the postharvest environment on the viability and
virulence of decay fungiCritical Reviews in Food Science and Nutrition58(10)1681168710.1080/10408398.2017.1279122
Maqbool, M., Ali, A., Alderson, P. G., Zahid, N. & Siddiqui, Y.
(2011). Effect of a Novel Edible Composite Coating Based on Gum Arabic and
Chitosan on Biochemical and Physiological Responses of Banana Fruits during Cold
Storage. Journal of Agricultural and Food Chemistry, 59(10),
5474-5482. DOI:10.1021/jf200623m.
M. Maqbool
A. Ali
P. G. Alderson
N. Zahid
Y. Siddiqui
2011Effect of a Novel Edible Composite Coating Based on Gum Arabic
and Chitosan on Biochemical and Physiological Responses of Banana Fruits
during Cold StorageJournal of Agricultural and Food Chemistry59(10)5474548210.1021/jf200623m
Martínez-Camacho, A. P., Cortez-Rocha, M. O., Ezquerra-Brauer, J.
M., Graciano-Verdugo, A. Z., Rodriguez-Félix, F., Castillo-Ortega, M.M.,
Yépiz-Gómez, M.S. & Plascencia-Jatomea, M. (2010). Chitosan composite films:
Thermal, structural, mechanical and antifungal properties. Carbohydrate
Polymers, 82(2), 305-315.
DOI:https://doi.org/10.1016/j.carbpol.2010.04.069.
A. P. Martínez-Camacho
M. O. Cortez-Rocha
J. M. Ezquerra-Brauer
A. Z. Graciano-Verdugo
F. Rodriguez-Félix
M.M. Castillo-Ortega
M.S. Yépiz-Gómez
M. Plascencia-Jatomea
2010Chitosan composite films: Thermal, structural, mechanical and
antifungal propertiesCarbohydrate Polymers82(2)30531510.1016/j.carbpol.2010.04.069
Mayek-Pérez, N., Pedroza-Flores, J. A., Villarreal-García, L. A.
& Valdés-Lozano, C. G. (1995). Factores genéticos y ambientales relacionados
con la dinámica temporal y efecto de las enfermedades en frijol
(Phaseolus vulgaris L.) en Marín, Nuevo León, México.
Revista Mexicana de Fitopatología, 13(1), 1-9.
N. Mayek-Pérez
J. A. Pedroza-Flores
L. A. Villarreal-García
C. G. Valdés-Lozano
1995Factores genéticos y ambientales relacionados con la dinámica
temporal y efecto de las enfermedades en frijol (Phaseolus vulgaris L.) en
Marín, Nuevo León, MéxicoRevista Mexicana de Fitopatología,13(1)19
Olicón-Hernández, D. R., Camacho-Morales, R. L., Pozo, C.,
González-López, J. & Aranda, E. (2019). Evaluation of diclofenac
biodegradation by the ascomycete fungus Penicillium oxalicum at
flask and bench bioreactor scales. Science of The Total
Environment, 662, 607-614.
DOI:https://doi.org/10.1016/j.scitotenv.2019.01.248.
D. R. Olicón-Hernández
R. L. Camacho-Morales
C. Pozo
J. González-López
E. Aranda
2019Evaluation of diclofenac biodegradation by the ascomycete fungus
Penicillium oxalicum at flask and bench bioreactor scalesScience of The Total Environment66260761410.1016/j.scitotenv.2019.01.248
Olicón-Hernández, D. R., Hernández-Lauzardo, A. N., Pardo, J. P.,
Peña, A., Velázquez-del Valle, M. G. & Guerra-Sánchez, G. (2015). Influence
of chitosan and its derivatives on cell development and physiology of
Ustilago maydis. International Journal of
Biological Macromolecules, 79, 654-660.
DOI:https://doi.org/10.1016/j.ijbiomac.2015.05.057.
D. R. Olicón-Hernández
A. N. Hernández-Lauzardo
J. P. Pardo
A. Peña
M. G. Velázquez-del Valle
G. Guerra-Sánchez
2015Influence of chitosan and its derivatives on cell development and
physiology of Ustilago maydisInternational Journal of Biological Macromolecules7965466010.1016/j.ijbiomac.2015.05.057
Olicón-Hernández, D. R., Uribe-Álvarez, C., Uribe-Carvajal, S.,
Pardo, J. P. & Guerra-Sánchez, G. (2017). Response of Ustilago
maydis against the Stress Caused by Three Polycationic Chitin
Derivatives. Molecules, 22(12), 1745.
D. R. Olicón-Hernández
C. Uribe-Álvarez
S. Uribe-Carvajal
J. P. Pardo
G. Guerra-Sánchez
2017Response of Ustilago maydis against the Stress Caused by Three
Polycationic Chitin DerivativesMolecules22(12)17451745
Olicón-Hernández, D. R., Zepeda Giraud, L. & Guerra-Sánchez, G.
(2017). Current applications of chitosan and chito-oligosaccharides.
Journal of Drug Design and Research, 4(2), 1039.
D. R. Olicón-Hernández
L. Zepeda Giraud
G. Guerra-Sánchez
2017Current applications of chitosan and
chito-oligosaccharidesJournal of Drug Design and Research4(2)10391039
Otálora, M. C., Carriazo, J. G., Iturriaga, L., Nazareno, M. A.
& Osorio, C. (2015). Microencapsulation of betalains obtained from cactus
fruit (Opuntia ficus-indica) by spray drying using cactus
cladode mucilage and maltodextrin as encapsulating agents. Food
Chemistry, 187, 174-181.
DOI:https://doi.org/10.1016/j.foodchem.2015.04.090.
M. C. Otálora
J. G. Carriazo
L. Iturriaga
M. A. Nazareno
C. Osorio
2015Microencapsulation of betalains obtained from cactus fruit
(Opuntia ficus-indica) by spray drying using cactus cladode mucilage and
maltodextrin as encapsulating agentsFood Chemistry18717418110.1016/j.foodchem.2015.04.090
Peris-Vicente, J., Marzo-Mas, A., Roca-Genovés, P., Carda-Broch, S.
& Esteve-Romero, J. (2016). Use of micellar liquid chromatography for rapid
monitoring of fungicides post harvest applied to citrus wastewater.
Journal of Environmental Sciences, 42, 284-292.
DOI:https://doi.org/10.1016/j.jes.2015.12.012.
J. Peris-Vicente
A. Marzo-Mas
P. Roca-Genovés
S. Carda-Broch
J. Esteve-Romero
2016Use of micellar liquid chromatography for rapid monitoring of
fungicides post harvest applied to citrus wastewaterJournal of Environmental Sciences4228429210.1016/j.jes.2015.12.012
Razavi, S. M. A., Mohammad Amini, A. & Zahedi, Y. (2015).
Characterization of a new biodegradable edible film based on sage seed gum:
Influence of plasticiser type and concentration. Food
Hydrocolloids, 43, 290-298.
DOI:https://doi.org/10.1016/j.foodhyd.2014.05.028.
S. M. A. Razavi
A. Mohammad Amini
Y. Zahedi
2015Characterization of a new biodegradable edible film based on sage
seed gum: Influence of plasticiser type and concentrationFood Hydrocolloids4329029810.1016/j.foodhyd.2014.05.028
Robledo, N., Vera, P., López, L., Yazdani-Pedram, M., Tapia, C.
& Abugoch, L. (2018). Thymol nanoemulsions incorporated in quinoa
protein/chitosan edible films; antifungal effect in cherry tomatoes.
Food Chemistry, 246, 211-219.
DOI:https://doi.org/10.1016/j.foodchem.2017.11.032.
N. Robledo
P. Vera
L. López
M. Yazdani-Pedram
C. Tapia
L. Abugoch
2018Thymol nanoemulsions incorporated in quinoa protein/chitosan
edible films; antifungal effect in cherry tomatoesFood Chemistry24621121910.1016/j.foodchem.2017.11.032
Saberi, B. & Golding, J. B. (2018). Postharvest Application of
Biopolymer-Based Edible Coatings to Improve the Quality of Fresh Horticultural
Produce. In T. J. Gutiérrez (Ed.), Polymers for Food
Applications (pp. 211-250). Cham: Springer International
Publishing.
B. Saberi
J. B. Golding
2018Postharvest Application of Biopolymer-Based Edible Coatings to
Improve the Quality of Fresh Horticultural Produce
T. J. Gutiérrez
Polymers for Food Applications211250ChamSpringer International Publishing
Seiber, J. N., Coats, J., Duke, S. O. & Gross, A. D. (2014).
Biopesticides: State of the Art and Future Opportunities. Journal of
Agricultural and Food Chemistry, 62(48), 11613-11619.
DOI:10.1021/jf504252n.
J. N. Seiber
J. Coats
S. O. Duke
A. D. Gross
2014Biopesticides: State of the Art and Future
OpportunitiesJournal of Agricultural and Food Chemistry62(48)116131161910.1021/jf504252n
Treviño-Garza, M. Z., García, S., Heredia, N., Alanís-Guzmán, M. G.
& Arévalo-Niño, K. (2017). Layer-by-layer edible coatings based on
mucilages, pullulan and chitosan and its effect on quality and preservation of
fresh-cut pineapple (Ananas comosus). Postharvest
Biology and Technology, 128, 63-75.
DOI:https://doi.org/10.1016/j.postharvbio.2017.01.007.
M. Z. Treviño-Garza
S. García
N. Heredia
M. G. Alanís-Guzmán
K. Arévalo-Niño
2017Layer-by-layer edible coatings based on mucilages, pullulan and
chitosan and its effect on quality and preservation of fresh-cut pineapple
(Ananas comosus)Postharvest Biology and Technology128637510.1016/j.postharvbio.2017.01.007
Valadez-Carmona, L., Cortez-García, R. M., Plazola-Jacinto, C. P.,
Necoechea-Mondragón, H. & Ortiz-Moreno, A. (2016). Effect of microwave
drying and oven drying on the water activity, color, phenolic compounds content
and antioxidant activity of coconut husk (Cocos nucifera L.).
Journal of Food Science and Technology, 53(9), 3495-3501.
DOI:10.1007/s13197-016-2324-7.
L. Valadez-Carmona
R. M. Cortez-García
C. P. Plazola-Jacinto
H. Necoechea-Mondragón
A. Ortiz-Moreno
2016Effect of microwave drying and oven drying on the water activity,
color, phenolic compounds content and antioxidant activity of coconut husk
(Cocos nucifera L.).Journal of Food Science and Technology53(9)3495350110.1007/s13197-016-2324-7
Viscometer, B. D. (2016). Operating Instructions. Manual
No. Manual No. M00-151-I0614. BROOKFIELD ENGINEERING LABORATORIES,
INC. 11 Commerce Boulevard, Middleboro, MA 02346 USA.
B. D. Viscometer
2016Operating Instructions. Manual No. Manual No.
M00-151-I0614BROOKFIELD ENGINEERING LABORATORIES, INC.Middleboro, MA
Younes, I. & Rinaudo, M. (2015). Chitin and Chitosan Preparation
from Marine Sources. Structure, Properties and Applications. Marine
Drugs, 13(3), 1133.DOI:10.3390/md13031133.
I. Younes
M. Rinaudo
2015Chitin and Chitosan Preparation from Marine Sources. Structure,
Properties and ApplicationsMarine Drugs13(3)1133113310.3390/md13031133
Yuan, G., Chen, X. & Li, D. (2016). Chitosan films and coatings
containing essential oils: The antioxidant and antimicrobial activity and
application in food systems. Food Research International, 89,
117-128. DOI:https://doi.org/10.1016/j.foodres.2016.10.004.
G. Yuan
X. Chen
D. Li
2016Chitosan films and coatings containing essential oils: The
antioxidant and antimicrobial activity and application in food
systemsFood Research International8911712810.1016/j.foodres.2016.10.004
Zambrano-Zaragoza, M., González-Reza, R., Mendoza-Muñoz, N.,
Miranda-Linares, V., Bernal-Couoh, T., Mendoza-Elvira, S. &
Quintanar-Guerrero, D. (2018). Nanosystems in edible coatings: A novel strategy
for food preservation. International Journal of Molecular
Sciences, 19(3), 705. DOI:10.3390/ijms19030705.
M. Zambrano-Zaragoza
R. González-Reza
N. Mendoza-Muñoz
V. Miranda-Linares
T. Bernal-Couoh
S. Mendoza-Elvira
D. Quintanar-Guerrero
2018Nanosystems in edible coatings: A novel strategy for food
preservationInternational Journal of Molecular Sciences19(3)70570510.3390/ijms19030705
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