Food and Nutrition Sciences, 2013, 4, 28-30
http://dx.doi.org/10.4236/fns.2013.47A004 Published Online July 2013 (http://www.scirp.org/journal/fns)
Qualitative Evaluation of the Capacity of Lactobacillus
Strains to Degrade Mycotoxins Developed and
Accumulated by Strains of the Genus Alternaria
L. Pérez, G. Girmé, E. L. Arosemena, M. A. Calvo Torras
Grupo de Investigación en Microbiología Aplicada y Medio-Ambiental, Departamento de Sanidad y de Anatomia Animales, Fa-
cultad de Veterinaria, Universidad Autónoma de Barcelona, Bellaterra, Spain.
Received March 27th, 2013; revised April 30th, 2013; accepted May 7th, 2013
Copyright © 2013 L. Pérez et al. This is an open access article distributed under the Creative Commons Attribution License, which
permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
ABSTRACT
Mycotoxins are secondary metabolites produced and accumulated by mould. This study assessed the ability of 89 lactic
acid bacteria, isolated from different sources, to degrade two mycotoxins by Alternaria: alternariol and altenuene. The
mehodology employed was the thin layer chromatography. The results obtained allow us to demonstrate that a 16.85%
of the strains (15/89 strains) are capable of decreasing the intensity of the band related at the alternariol and 14.60%
(13/89 strains) is capable of decrease the intensity of the band that manifests the altenuane in Alufolium. All tests have
been observed under wavelengths of 360 nm and 255 nm. The results obtained allow carrying out a selection of lactic
acid strains capable of degrading or decreasing the concentration of mycotoxins selected for this.
Keywords: Lactic Acid Bacteria; Mycotoxins; Alternaria; Thin Layer Chromatography
1. Introduction
Mycotoxins are compounds resulting from the condensa-
tion reactions which take place under certain physical,
chemical and biological conditions when the reduction of
ketone groups in the biosynthesis of fatty acids made by
the moulds is interrupted. These acids are primary me-
tabolites by the fungus used as energy source. Mycotox-
ins are often formed at the end of the exponential phase
or early stationary growth phase of the microorganism.
These are pollutants of various food and feed, corre-
lating with various diseases of animals and humans [1,2].
Exposure to mycotoxins can cause acute or chronic tox-
icity, and adverse effects on the central nervous system,
on the cardiovascular, respiratory and digestive system
and can also cause death. Agents may also be carcino-
genic, mutagenic, teratogenic and immunosuppressive.
At present it is mentioned as one of the most important
effects, the ability of some mycotoxin compromise the
immune response thus reduce the resistance to infectious
diseases.
Moreover mycotoxins are of global concern due to
major economic losses because of its effects at the level
of human health, animal productivity and national and
international trade. Miller [3] in a statement staff esti-
mate that in the United States of America and Canada,
the annual losses due to the effects of mycotoxins in feed
and livestock industries are of the order of 5000 million.
For prevention, decontamination, inactivation and detoxi-
fication have studied different methods: physical, chemi-
cal, fungistatic utilization, use of additives and enzymes
mycotoxin absorbents.
The latter method is applicable to the degradation and/
or inactivation of mycotoxins, which can include: natural
extracts and/or essential oils from plants and pathogenic
microorganisms.
Various tests have been conducted to study pathogenic
microorganisms capable of degrading specific conditions
certain mycotoxins. Such microorganisms are: Saccharo-
myces cerevisiae, Flavobacterium aurantiacum, Rhizopus
spp. Neurospora sitophila and rumen microorganisms.
Some of these studies have provided effective results
in the degradation of mycotoxins such as Aflatoxins, Pa-
tulin, Ochratoxin A, Zearalenone, T-2, Rubratoxin or Dia-
cetoxyscirpenol A. These studies have been performed in
the laboratory, but the practical application of these sys-
tems is under study and development [4].
Lactic acid bacteria (LAB) may be a possible alterna-
tive. This group of microorganisms is considered GRAS

Qualitative Evaluation of the Capacity of Lactobacillus Strains to Degrade Mycotoxins Developed
and Accumulated by Strains of the Genus Alternaria
29
(Generally Recognized As Safe) and are therefore ideal
for use as biocontrol or as biopreservants. Bacteria of the
genus Lactobacillus are widely distributed in nature and
grow in a variety of conditions [5,6], during their devel-
opment released into the metabolic products such as lac-
tic acid, acetic acid, hydrogen peroxide, bacteriocins,
among others [7] which can cooperate effectively to con-
trol undesirable microorganisms.
2. Materials and Methods
From 89 axenic strains of Lactobacillus, of various ori-
gins, as detailed in Table 1, and developed overnight in
MRS broth was obtained. An aliquot was inoculated into
50 mL of MRS broth that is calculated according the
proportion of 5% (v:v) corresponding to the final vol-
ume.
After inoculation of each of these cultures were ali-
quoted kinetics according to exponential phase growth of
these strains, determined between 0 - 7 hrs of incubation
[8].
During 30 minutes maintaining the interaction be-
tween each of the respective cultures mycotoxins and
filtered under study. The minimum concentration of my-
cotoxins is 0.020 ppm [9].
Interaction elapsed 10 μl are obtained, which are de-
posited in silica gel chromatoplates (Ref: 0554.001, Merck).
Previously these chromatoplates were activated at 100˚C
for 15 minutes [9].
Positive controls:
 10 μl alternariol (M1) + 10 μl MRS Broth;
 10 μl altenuene (M2) + 10 μl MRS Broth.
Negative controls:
 Filtering 10 μl of strain + 10 μl strain MRS Broth;
 10 μl cultivation of strain + 10 μl unfiltered MRS
Broth.
Mobile phase was used as the following mixture: tolu-
ene/methanol/acetic acid (86:12:2) [9].
After the chromatography, the chromatoplates were
observed under uv at 365 nm and 255 nm. The fluores-
cence bands of samples and controls problems allow us
to compare the values of Rf and set in the semiquantita-
tive results.
3. Results and Discussion
Several authors have shown in their work that secondary
metabolites produced and accumulated by strains of the
genus Alternaria are toxic [10-12]. In the course of our
work we have selected two of the genus Alternaria my-
cotoxins: alternariol and altenuene, to conduct studies on
the potential degradation of mycotoxins by the 89 strains
of lactic acid bacteria that have submitted ability to in-
hibit Fusarium moniliforme and/or Alternaria alternata.
The results obtained allow us to demonstrate that a
16.85% (15/89) are capable of reducing the intensity of
the band at alternariol observed at wavelengths 360 nn
and 255 nn and 14.60% (13/89) is capable of decreasing
the intensity of the band expressed by altenuene observed
at wavelengths 360 nm and 255 nm.
The Figure 1 shows chromatoplate, showing de-
creased intensity of the band representing the mycotoxin.
Decrease is attributed to the interaction of the mycotoxin
with lactic strains
This technique is relatively fast but only qualitative re-
sults can be achieved, thus allowing not deliver numeri-
cal results representing the decrease in concentration of
the mycotoxin. However, facilitates carrying out a selec-
tion of strains BAL, with manifest ability to degrade or
reduce the concentration of the two mycotoxins selected
for this study: alternariol and altenuene.
4. Conclusion
The two mycotoxins used as substrates for degradation
capacity tests, were modified or degraded by 24 strains
Table 1. Number of strains tested and origin.
Origin of Lactobacillus strains Number of strains tested
Small intestine and caecum of birds 8
Dog faeces 29
Pig faeces 41
Snails faeces 11
Figure 1. Testing thin layer chromatography mycotoxin re-
duction bands: A: interaction of M1 with strain C-P11; B:
M2 interaction with strain C-C50; C: interaction with the
M1 and M2 strain N-C61; C: Filtered N: Strain unfiltered;
M1: M2 alternariol: altenuene.

Qualitative Evaluation of the Capacity of Lactobacillus Strains to Degrade Mycotoxins Developed
and Accumulated by Strains of the Genus Alternaria
30
of lactic acid bacteria, of which, fifteen strains showed
activity on alternariol and thirteen on altenuene.
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