Wageningen Academic

World Mycotoxin Journal, May 2008; 1(2): 139-146 P u b l i s h e r s

Screening of antifungal and antimycotoxigenic activity of plant phenolic extracts

M. dos Santos Oliveira and E. Badiale Furlong

Fundação Universidade Federal do Rio Grande (FURG), Departamento de Química Laboratório de Micotoxinas, Rua
Eng°Alfredo Huch 475, CEP 96201-900 Rio Grande/RS, Brazil; dqmebf@furg.br

Received: 21 November 2007 / Accepted: 9 February 2008

© 2008 Wageningen Academic Publishers

Abstract

The antifungal and antimycotoxigenic activities of extracts from edible plants were tested by the agar dilution
method using the growth diameter of Aspergillus flavus as response and the determination of aflatoxins B1 and B2
in the culture medium. On the 7th incubation day, the greatest fungal inhibitions were reached by the extracts from
potato peel; rice and wheat; lemon peel and pulp; eggplant peel; orange peel and pulp; and apple pulp. After the
14-day incubation, the extracts from banana (30 µg phenol/ml agar), eggplant (30 µg phenol/ml agar), and potato
(50 and 67 µg phenol/ml agar) pulp reduced the production of aflatoxin B1 by 3.2%/µg phenol/ml agar, 2.9%/µg
phenol/ml agar, 1.8%/µg phenol/ml agar and 0.85%/µg phenol/ml agar, respectively, in relation to the control. The
extracts from the other vegetables fully inhibited the synthesis of the mycotoxin. These results point to the studied
plants and their residues as potential sources of phenolic compounds that may have an inhibitory effect on fungal
development and the production of mycotoxins in food.

Keywords: phenolic compound, Aspergillus flavus, natural antifungal

1. Introduction Brazil (Bittencourt et al., 2005; Garda et al., 2004; Nunes

In food production it is always necessary to seek appropriate
strategies for ensuring product stability during their shelf
life and their microbiological safety, particularly in regard
to fungal contamination (Brul and Coote, 1999). Food
contamination by fungi leads to changes in chemical
composition, structure and appearance, thus representing
economic loss and/or waste of raw material and food. In
addition, the fungi are responsible for health problems in
both humans and animals, if they ingest food contaminated
by mycotoxins produced by toxigenic species in stressful
conditions (Taniwaki and Silva, 2001; Paster et al., 1999).
Mycotoxins are secondary metabolites of certain strains
of toxigenic fungi that have been shown to be toxigenic,
carcinogenic, mutagenic, neurotoxic, and teratogenic to
different species of animals. These mycotoxin-producing
fungi are widely distributed in nature and can grow in a
wide range of environmental conditions. Their occurrence
in food has been cited in different countries, including
et al., 2003; Aycicek et al., 2005; Abdulkadar et al., 2004).
The mycotoxigenic fungi involved in the human food
chain belong mainly to genera Aspergillus, Penicillium,
and Fusarium (Sweeney and Dobson, 1998).
The aflatoxins, of which five are of concern as natural
contaminants in food (aflatoxin B1, B2, G1, G2 and M1),
are secondary metabolites from Aspergillus flavus and A.
parasiticus. Aflatoxins have been detected in foods, feed
and agricultural commodities, in different concentrations.
Their toxic effects may manifest chronically (cancer) or
acutely (diarrhoea, vomiting, liver disorders) (Sweeney and
Dobson, 1998; Moss, 2002).
Strategies for the prevention of mycotoxigenic fungus
growth and the further production of mycotoxins in
agricultural produce and food are the use of fungicide
and chemical preservatives or heat treatment to inactivate
spores (Yoshizawa, 2001). The resistance of fungi to

ISSN 1875-0710 print, ISSN 1875-0796 online 139

M. dos Santos Oliveira and E. Badiale Furlong
chemical preservatives and legal limitations ensure an
increasing demand for naturally preserved products.
The search for natural sources of antifungal additives that
are safe and efficient to be used in food has shown that
extracts and essential oils from spices, herbs, and other
plants carry antifungal activity (Soliman and Badeaa, 2002;
Juglal et al., 2002; Fan and Chen, 1999). Some compounds
have been cited as being capable of inhibiting toxigenic
species growth, as well as mycotoxin production (Sánchez
et al., 2004; Gowda et al., 2004; Marín et al., 2004). Evidence
suggests that phenolic compounds naturally found in
plants, herbs, fruit, cereals, and other vegetables, as well
as their essential oils, carry antifungal activity, and may
control mycotoxin production (Rasooli and Abyaneh, 2004;
Bakan et al., 2003). The results may be a way to recover
waste or material rejected from domestic or industrial
routine processes.
Several phenolic structures, such as carnosol and eugenol,
are found in herbs and spices; flavonoids, as well as
chlorogenic, cafeic, ferulic, and cinnamic acids are found
in fruit (Robards et al., 1999). Fungal inhibition has been
reported by phenolic compounds such as eugenol over
Penicillium species (Venturini et al., 2002; Vásquez et al.,
2001), as well as antifungal activity by phenolic compounds
found in orange (Del Río et al., 1998), pepper essential oil
(Ejechi et al., 1999), olive trees (Del Río et al., 2003), and
other edible or medicinal plants (Rauha et al., 2000).
Such considerations have guided the objective of the
present study, which was to investigate the antifungal and
antimycotoxigenic activity of phenolic extracts from edible
plant which has been used in daily diets in many regions
of the world.
2. Material and methods
Plant material
The plants studied included orange (Citrus sinensis,
Osbeck), lemon (Citrus limon), apple (Malus domestica
Borkh), banana (Musa spp.), rose potato (Solanum
tuberosum L.), eggplant (Sonalum melongina, L.), processed
rice (Oryza sativa), and whole wheat (Triticum aestivum),
purchased in a market in the South of Brazil. Fruit and
tubercle peel were separated from their edible portions,
and then cut into 5 mm cubes. Cereal grains were milled
using a knife-mill, down to 0.42 mm grain size.
Total phenolic assay
Phenol compounds were cold-extracted with methyl
alcohol at a 1:5 dry mass:solvent ratio as described by
Badiale-Furlong et al. (2003), then quantified using the
spectrophotometric method with Folin-Ciocalteau’s
140
reagent and a tyrosine standard curve ranging between 0
and 5 µg/ml. The extracts were concentrated under vacuum
for complete removal of solvents. The solvent-free extracts
were used for the present study.
Antifungal activity test
The toxigenic filamentous fungus used in the present
study was Aspergillus flavus, supplied by Laboratório de
Microbiologia at FURG’s Chemistry Department, and kept
in Potato Dextrose Agar (PDA) at 4 °C. The inoculum was
prepared by growing the fungi on PDA at 30 °C for 10 days.
Spores were harvested by washing the agar surface with
a sterile solution containing 0.2% Pure Agar in Tween 80.
Spore counts in the resulting suspensions were determined
by microscopy with Neubauer chamber.
Antifungal activity was determined by agar dilution
method. The test plant extract was added to PDA Agar in
two different volumes, completing two different levels of
total phenol at growth medium, called level 1 and level 2.
In the control, the same amount of sterile water was added.
Using a micropipette, an inoculum of 5 µl of the spore
suspensions (107 spores/ml) was added to the centre of
Petri dish. The cultures were incubated at 30 °C for 7 days
and the temperature was then reduced to 21 °C for 7 days.
The radial growth of mycelia was measured on the 3rd, 5th,
7th, 10th, and 14th day of incubation for the experiments,
conducted as triplicates (Nguefack et al., 2004). After 14
days, dishes were frozen (-10 °C) for further determination
of mycotoxins in the growth medium.
Evaluation of aflatoxins B1 and B2 production
Mycotoxins were extracted from the culture media with an
acetonitrile-water (3:1) solution while stirring, followed by
hexane-based partition. Acetonitrile was separated from
the water by addition of sodium chloride, followed by
vacuum-evaporation at 50 °C. The residue was resuspended
with methanol:chloroform 1:9 (v/v), and centrifuged.
Three 10 ml aliquots were separated in an amber glass.
They were evaporated in a water-bath under nitrogen
current and stored at -10 °C (Tanaka et al., 2001). The
evaporated extracts were applied to 0.25 mm G60 silica
gel chromatosheets for screening and quantification of
aflatoxins B1 and B2 under UV light (Soares and Rodriguez-
Amaya, 1989).
Statistical analysis
Analysis of variance (ANOVA) was performed on all
data using the Statistic Software version 5.1. The mean
values of development diameters were compared by the
least significant difference (LSD) test at a 5% level of
confidence.
World Mycotoxin Journal 1 (2)
 Screening of antifungal and antimycotoxigenic activity of plant phenolic extracts
3. Results and discussion
Medicinal herbs, spices (oregano, clove, cinnamon,
pepper), and other edible or non-edible plants are usually
studied as their antifungal vegetable compounds. Some
researchers have shown that extracts and essential oils from
garlic and onions were able to inhibit the development of
Aspergillus, Fusarium, and Penicillium species (Yin and
Tsao, 1999; Benkeblia, 2004). However, the fruits lemon,
orange, banana, eggplant, apple; the tubercle potato; and
the grains rice and wheat, which are commonly found in
human diets around the world, have not been thoroughly
studied regarding this ability. In the present study, they
were evaluated on the amounts of total phenolic in their
edible part, or pulp, and peels (Table 1). The use of such
food in distinct regions of the country without an effective
use for their residues prompted the interest of the research
in these vegetables.
The total phenolic compounds content in the edible portion
of the vegetable tissues studied ranged from 40 µg phenol/g
(dry base) in rice to 4,500 µg phenol/g (dry base) in orange
pulp. The phenolic content in peels ranged from 990 µg
phenol/g (dry base) in potato peel to 5,500 µg phenol/g (dry
base) in lemon peel. The total phenolic content in lemon,
apple, banana, eggplant and potato peels was higher than in
their respective pulps. The opposite behaviour was found
in orange peel.
Phenolic extract effect on the growth of Aspergillus flavus
The effect of the extracts was evaluated by measurements
of the growth diameter of the colonies. Every tested plant
extract provided inhibition to the growth of A. flavus in
both concentrations used. An upward linear trend was
found for the relationship between the concentration and
the inhibiting effect for the same extract.
Table 1. Total phenol content in vegetable portions.
Edible portion Total phenolic compounds
µg/g (dry base)
lemon pulp 2,700±57a
orange pulp 4,500±52
apple pulp 1,000±2.6
banana pulp 310±8.5
eggplant pulp 390±5.8
potato pulp 410±14
rice 40±1.1
wheat 290±9.5
a Mean±Standard deviation.
World Mycotoxin Journal 1 (2)
Table 2 shows the assessment results of the Aspergillus
flavus growth diameter in the presence of phenolic extracts
from vegetable peels (lemon, orange, apple, banana,
eggplant, and potato) on the 3rd, 5th, 7th, 10 th and 14th day
of inoculation. On the 7th incubation day, colony diameters
measured from 0 cm for mediums containing potato peel
extract (38 µg phenol/ml agar) to 5.9 cm for mediums
containing banana peel extract (44 µg phenol/ml agar);
on the 14th day, such measures were from 0 cm to 6.7 cm
for the same extracts.
The percentage of inhibition by the extracts was estimated
relating the growth diameter of the colony developed in the
medium with the extracts and absence of them. The potato
peel extracts that presented lower phenolic compound
concentration and the colony diameter estimation
showed 3%/µg phenol/ml agar and 2.5%/µg phenol/ml
agar inhibition. Such profile was maintained until the 14th
incubation day for the higher phenolic concentration level
(40µg phenol/ml agar). This high inhibition effect did not
take place with the other phenolic extract from peels with
higher phenol rates, like lemon, orange and apple.
The results indicate that the type of phenolic structure is
more important than the concentration. It is known that
some fungi are able to adapt in the presence of some classic
antifungal compounds, but this is not observed in the case
of phenolic extracts from potato peel. Hydroxyl groups
assigned to the phenolic compounds may form hydrogen
bridge bonds with active enzymes, resulting in their
deactivation and a consequent effect on the development
of the fungal biomass and the production of mycotoxins
(Juglal et al., 2002).
The mean comparison by least significant differences
conducted for each day of growth diameter measurements
showed that on the 3rd and 5th days of incubation, the colony
Residue Total phenolic compounds
µg/g (dry base)
lemon peel 5,500±160
orange peel 4,200±37
apple peel 4,700±140
banana peel 1,010±25
eggplant peel 2,200±96
potato peel 990±38
141
M. dos Santos Oliveira and E. Badiale Furlong

Table 2. Mean growth diameters of Aspergillus flavus in the presence of vegetable peel extracts.

Peel extracts Total phenol Colony diameter (cm)

µg/ml agar
3rd day 5th day 7th day 10th day 14th day

banana 44 1.2±0.5 3.0±0.6 6.0±1.0 6.5±1.0 6.7±0.4

banana 58 0.9±0.14 1.8±0.6 3.0±0.6 5.1±1.0 6.3±0.5
lemon 210 1.0±0.1 1.4±0.5 2.6±0.2 3.0±0.2 3.6±0.3
lemon 370 0 0.2±0 0.4±0.04 0.4±0.07 0.7±0.02
orange 140 1.1±0.03 1.8±0.07 3.0±0.1 3.6±0.09 4.4±0.1
orange 250 0.2±0.03 0.8±0.1 1.4±0.3 1.6±0.4 2.2±0.5
apple 130 1.2±0.3 3.2±0.3 4.6±0.8 5.2±0.6 6.3±0.3
apple 200 1.0±0.5 2.0±0.8 2.8±0.7 4.5±0.3 4.7±0.3
eggplant 50 0.7±0 1.3±0.2 3.0±0.5 3.6±0.7 4.7±0.5
eggplant 60 0 0.6±0.06 0.8±0.05 0.9±0.03 1.6±0.7
potato 30 0 0.7±0.1 0.8±0.04 1.3±0.3 2.2±0.9
potato 38 0 0 0 0 0
control - 4.6±0.1 6.5±0.03 6.6±0.2 6.7±0.3 7.6±0.3

diameters in the presence of extracts were significantly
smaller (74%) relative to the control, confirming the
reduction in A. flavus growth rates. On the 7th incubation
day, only when banana peel extract (44 µg phenol/ml agar)
was used, it was found that fungus growth was proximate
to the control. From the 10th incubation day on, banana
peel extracts (44 and 58 µg phenol/ml agar) did not provide
any inhibition, and the colony diameters did not present a
significant difference relative to controls. Such behaviour
may have been determined by the type and concentration
of the phenolic compounds present in the extract, which
was not enough to inhibit the initial amount of spores
present in the growth medium.
It is reported in literature that concentrations between
3,500 and 5,000 µg phenolic acids/ml growth medium,
extracted from pepper, are the minimum concentrations
required to fully inhibit fungi Rhyzopus stolonifer, A. niger,
and Fusarium spp. after 48 hours (Ejechi et al., 1999).
Figure 1 illustrates the inhibitory effects of vegetable
peel extracts, added in two distinct levels, upon the
development of A. flavus with 7 days of incubation. The
highest inhibition on the 7th incubation day was found in
the medium containing potato peel extracts (level 2), with
100% inhibition, followed by lemon peel extracts (level 2)
with 96%, eggplant peel (level 2) with 90%, and potato
peel extract (level 1) with 88%. On the 14th incubation day
(Figure 2), the same performance sequence was maintained.
The extracts presented respectively 100%, 91%, 79%, and
71% of fungal growth inhibition.
1 and 2 show that the greatest inhibition occurred when
extracts with the highest phenolic rates were used. The
LSD test indicated that on the 7th incubation day, there was
a significant difference between inhibition occurring for
the lower and higher phenol ratios upon colony diameter,
with the exception of potato peel extract. At 14 days of
incubation, there was no significant difference between
the inhibition caused by the higher phenolic compounds
concentration and the lower concentration of banana and
apple peel extracts.
Table 3 shows growth diameter results for A. flavus
inoculated in the presence of phenolic compounds
extracted from vegetable pulps on the 3rd, 5th, 7th, 10th, and
14th day of incubation. On the 7th incubation day, colony
diameters ranged between 0 cm for rice (6 µg phenol/ml
agar) and wheat (9 µg phenol/ml agar) extract mediums,
and 4.2 cm for orange pulp extracts (110 µg phenol/ml
agar). On the 14th day, these scores ranged from 0 cm to
6.4 cm for mediums containing the same extracts. The
inhibitory power of rice and wheat extracts stood out,
considering that they presented the lowest phenol rates.
The variance analysis showed a significant difference (95%)
between growth diameters in the presence of pulps. The
LSD test indicated that on the 3rd, 5th, and 7th days of
incubation, extract inhibition was significant relative to
the control. On the 10th and 14th incubation days, only the
orange pulp extract added to the lower phenol level did not
maintain inhibition, and colony diameters did not present
significant differences relative to the control.

Variance analysis indicated that there was a significant In general, pulp extract also provided inhibition of the
difference, with 95% confidence, among growth diameter development rate of A. flavus, keeping its growth diameter
means in the presence of the distinct plant extracts. Figures significantly smaller than the control test until the 14th

142 World Mycotoxin Journal 1 (2)

 Screening of antifungal and antimycotoxigenic activity of plant phenolic extracts

100 b
a
100 b
b a b
80 b
80 b
b a
Inhibition (%)

Inhibition (%)
a
60 b a b a 60 a
a
40 40 a a
a

20 20 a a
a a
Level 1 0 Level 1
0 Level 2
banana lemon orange apple eggplant potato Level 2 banana lemon orange apple eggplant potato

Figure 1. Inhibitory effect of vegetable peel extracts upon
the growth of Aspergillus flavus during 7 days of incubation.
Level 1: Lower phenolic extract level from each vegetable
added to the growth medium; Level 2: Higher phenolic extract
level from each vegetable added to the growth medium. a,b:
The same letters in the extract indicates that there are no
differences between the inhibition exerted by the different
phenolic concentrations.
Figure 2. Inhibitory effect of vegetable peel extracts upon
the growth of Aspergillus flavus after 14 days of incubation.
Level 1: Lower phenolic extract level from each vegetable
added to the growth medium; Level 2: Higher phenolic extract.
a,b: The same letters in the extract indicates that there are
no differences between the inhibition exerted by the different
phenolic concentrations.

Table 3. Radial growth of Aspergillus flavus in the presence of vegetable pulp extracts.

Pulp extracts Total phenol Colony diameter (cm)

µg/ml agar
3rd day 5th day 7th day 10th day 14th day

banana 30 1.0±0.4 2.0±0.5 3.8±1.0 4.6±1.0 5.8±1.4

banana 40 1.2±0.3 2.2±0.9 3.8±1.6 4.7±0.8 5.5±0.7
lemon 110 1.1±0.2 2.0±0.5 3.4±0.4 4.1±0.3 4.9±0.4
lemon 170 0.3±0.02 0.7±0.03 1.7±0.9 1.9±0.9 2.4±0.2
orange 110 1.2±0.2 2.2±0.3 4.2±0.7 5.3±0.6 6.4±0.6
orange 170 0.3±0.02 1.2±0.3 2.4±0.5 3.2±0.7 4.4±0.5
apple 67 0.8±0.1 1.6±0.3 2.8±0.9 4.2±1.0 5.1±1.0
apple 88 0.3±0.02 0.6±0.07 1.2±0.3 1.8±0.5 2.6±0.6
eggplant 30 0.9±0.2 1.4±0.4 3.0±0.7 3.9±0.8 5.1±0.8
eggplant 40 0.7±0.02 1.4±0.4 3.1±0.6 3.8±0.9 4.6±1.0
potato 50 0.7±0.05 1.9±0.4 3.8±0.7 4.8±0.7 5.1±0.4
potato 67 0.8±0.2 1.9±0.9 3.3±0.6 4.3±0.7 4.8±0.5
rice 4.5 0 0.4±0.02 0.8±0.05 1.6±0.06 1.9±0.07
rice 6 0 0 0 0 0
wheat 7 0 0.2±0 0.5±0 1.0±0.05 1.6±0.07
wheat 9 0 0 0 0.3±0.06 0.9±0.03
control 4.6±0.1 6.5±0.03 6.6±0.2 6.7±0.3 7.6±0.3

day of incubation, except for the orange pulp. Figures 3
and 4 present fungal inhibition effects of phenol extracts
contained in vegetable pulp upon the development
of A. flavus on the 7 th and 14 th days of incubation,
respectively.
also seen in the presence of rice and wheat extracts. The
LSD test (confidence level 95%) showed that there was no
significant difference between the lower and higher phenol
levels in banana, eggplant, potato, rice, and wheat pulp
extracts added to the growth medium.

On the 7th incubation day, the greatest inhibition occurred Effects presented by peel extracts which caused the greatest
in the presence of rice, wheat and apple pulp extracts. After inhibitions such as lemon, potato, eggplant, and orange and
14 days of incubation (Figure 4), the greatest inhibition was the best pulp extracts, such as rice, wheat, apple, lemon and

World Mycotoxin Journal 1 (2) 143

M. dos Santos Oliveira and E. Badiale Furlong

100 a a 100 b
a
a a
b a
80 b 80 a
b b
b
Inhibition (%)

Inhibition (%)
60 a a a 60
a a
a a a b a
40 a 40 a a a
a
a
a
a
20 20 a

0 Level 1 Level 1
banana lemon orange apple eggplantpotato rice wheat Level 2 0
banana lemon orange apple eggplant potato rice wheat Level 2

Figure 3. Effect of extracts from vegetable tissue pulp upon
the growth of Aspergillus flavus during 7 days of incubation.
Level 1: Lower phenolic extract level from each vegetable
added to the growth medium; Level 2: Higher phenolic extract.
a,b: The same letters in the extract indicates that there are
no differences between the inhibition exerted by the different
phenolic concentrations.
Figure 4. Effect of extracts from vegetable tissue pulp upon
the growth of Aspergillus flavus after 14 days of incubation.
Level 1: Lower phenolic extract level from each vegetable
added to the growth medium; Level 2: Higher phenolic extract.
a,b: The same letters in the extract indicates that there are
no differences between the inhibition exerted by the different
phenolic concentrations.

orange, were subject to variance analysis (95%), resulting of 2 to 20 µg afla/kg agar with a variation coefficient of 8%
in a significant difference between inhibitions caused by for both mycotoxins.
such extracts. It is worth emphasizing that, after 7 days
of incubation, the greatest fungal inhibition varied from An inhibitory effect upon the production of aflatoxin B1
100% when potato, rice and wheat shells were used with and B2 by A. flavus was seen in the presence of every
the higher phenolic level, to 64%, using orange pulp extract extract at both studied concentrations. It is interesting to
with the higher phenolic level (170 µg phenol/ml agar). note that the aflatoxin B1 and B2 production inhibition
After 14 days, the greatest inhibitions ranged from 100% was relatively higher than the inhibition of A. flavus
to 66%. Such vegetable tissues were the best of the ones development, when developed in the presence of extracts
studied as potential sources of phenolic compounds, and from orange, lemon, apple, rice, wheat pulp and peel, and
could be employed as antifungal additives. Fungal inhibition banana and eggplant peel.
was highest with the use of rice, wheat, potato peel, lemon
and orange peel and pulp, eggplant peel, and apple pulp. Extracts from vegetable peels and 62.5% of pulp extracts
fully inhibited the production of aflatoxin B 1 and B2;
Phenolic extract effect on the production of aflatoxin B1
and B2 by Aspergillus flavus.
Table 4. Aflatoxin B1 and B2 production by Aspergillus flavus
The most important mycotoxins produced by A. flavus are in the presence of phenolic compounds from vegetable
aflatoxins B1 and B2, formed by about 40% of the isolated extracts.
fungi (Taniwaki and Silva, 2001). Table 4 shows the results
of determinations of aflatoxins B1 and B2 produced by A. Extracts present of agar Quantification (µg/kg agar)
flavus in Potato Dextrose Agar medium in the presence
and absence of phenolic extracts from the vegetables. The aflatoxin B1 aflatoxin B2
inhibition rate was calculated considering the absence of
production mycotoxin as 100% inhibition. control 1,600±140 (0)a nd
banana peel (44µg phenol/ml agar) nd nd
The method by Tanaka (2001) and Soares and Rodriguez- orange peel (250µg phenol/ml agar) nd nd
Amaya (1989), adapted at FURG’s Laboratório de banana pulp (30µg phenol/ml agar) 75±7.1 (95) <100±0
Micotoxinas for determination of mycotoxins was adapted eggplant pulp (30µg phenol/ml agar) 195±7.1 (87) nd
because it is fast, economic and has a good performance to potato pulp (50µg phenol/ml agar) <100±2.1 (94) <140±7.1
evaluate mycotoxin production in the culture medium. The potato pulp (67µg phenol/ml agar) 670±120 (57) 850±200
method has a detection limit of 2 µg afla/kg agar both for
aflatoxin B1 and B2. The recoveries for aflatoxin B1 and B2 nd = not detected (detection limit 2 µg/kg agar).
are 96.5% and 91.5% respectively at a concentration range a Percentage inhibition between brackets.

144 World Mycotoxin Journal 1 (2)

 Screening of antifungal and antimycotoxigenic activity of plant phenolic extracts
whereas extracts from vegetable peels on the 14th incubation
day presented fungal inhibition ranging from 12% in the
presence of banana peel extract to 100%, when potato peel
extract was used. During the same period, pulp extracts
presented 16-100% inhibition of fungal colony development.
When used, banana pulp (30 µg phenol/ml agar), eggplant
pulp (30 µg phenol/ml agar), and potato pulp (50 and 67 µg
phenol/ml agar) extracts inhibited respectively 3.2%/µg
phenol/ml agar, 2,9%/µg phenol/ml agar, 1.88%/µg phenol/
ml agar and 0.85%/µg phenol/ml agar of the aflatoxin B1
production (Table 4), while fungal inhibition was 0.80%/µg
phenol/ml agar, 1.1%/µg phenol/ml agar, 0.66%/µg phenol/
ml agar, and 0.58%/µg phenol/ml agar for the same extracts
on the 14th day of incubation. Such phenolic extracts may
act by inhibiting oxidative stress, which is a prerequisite
for the production of aflatoxins, leading to an increase in
the lipidic peroxidation and the generation of free radicals
(Jayashree and Subramanyam, 2000). Similar results were
found by Selvi et al. (2003), who inoculated Aspergillus
flavus in the presence of extract of Garcinia indica, a plant
rich in flavonoids and phenolic acids. The authors obtained
a 40-60% inhibition of the production of aflatoxin B1 and
a 10-20% inhibition of fungal growth.
It is worth emphasizing that in the presence of banana
pulp (30 µg phenol/ml agar) and potato pulp (50 and 67 µg
phenol/ml agar) extracts, A. flavus also produced aflatoxin
B2, which was not found for the control. Such secondary
production may be a defence response of the fungus when
in the presence of a component of those extracts. The
results indicated that the antifungal and antimycotoxigenic
activities of vegetable extracts were related to the type of
phenolic compounds found in the system, a fact which
has been reported by other studies. These possibilities are
being studied in our laboratory.
Bakan et al. (2003) studied the effect of natural substrates in
three fractions of corn, whole grains, degerminated grain,
and wheat germen upon the production of thrichothecenes
by Fusarium graminearum. They also found that the
production of mycotoxins was lower or undetectable in
the wheat germen, a fraction which presented the highest
concentration of ρ-coumaric and ferulic acids.
4. Conclusions
Every extract presented antifungal activity on A. flavus. Full
inhibition after 7 days of incubation was reached by potato
peel (38 µg phenol/ml agar), rice (6 µg phenol/ml agar),
and wheat (9 µg phenol/ml agar) extracts. By 14 days of
incubation, full inhibition was found for potato peel (38 µg
phenol/ml agar) and rice (6 µg phenol/ml agar) extracts.
Vegetable peel extracts and some pulp extracts (62.5% of
then) inhibited aflatoxin production at 100%. Banana pulp
(30 µg phenol/ml agar), eggplant pulp (30 µg phenol/ml
agar), and potato pulp (50 and 67 µg phenol/ml agar)
World Mycotoxin Journal 1 (2)
extracts inhibited the production of aflatoxin B1 at 3.2%/µg
phenol/ml agar, 2.9%/µg phenol/ml agar, 1.88%/µg phenol/
ml agar and 0.85%/µg phenol/ml agar, respectively.
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