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ScienceDirect
Biosensors for early detection of fungi spoilage and
toxigenic and mycotoxins in food
Idjane Santana Oliveira1, Alberto Galdino da Silva Junior2,
Cesar Augusto Souza de Andrade2 and
Maria Danielly Lima Oliveira2
Biosensors represent technology that can be applied to several
sectors of the food industry from the storage of grains and raw
materials, food production/processing, security and
protection, packaging of food. Diverse biosensors have
emerged in the last decade as an alternative for analyzing
microorganisms and toxins in food due to the capability for fast
analysis, reproducibility, stability, and accuracy. A wide variety
of transducers can be explored for mycotoxin and spoilage,
fungi detection, where optical (surface plasmon resonance –
SPR and fluorescence), piezoelectric (quartz crystal
microbalance – QCM), and electrochemical (impedimetric,
potentiometric and amperometric) spectroscopies stand out as
main biosensing method. Biological materials (e.g. peptides,
enzymes, antibodies, cells, nucleic acids) and bioinspired tools
such as aptamers, molecularly imprinted polymers (MIPs) and
recombinant antibodies present a high focus as sensing
elements on biosensor studies Herein, we present an overview
of the recent progress of the development of new biosensors
for early detection of fungi (spoilage and toxigenic) and
mycotoxins in food.
Addresses
1
Centro Acadêmico de Vitória, Universidade Federal de Pernambuco,
55.608-680, Vitória de Santo Antão, PE, Brazil
2
Departamento de Bioquı́mica, Universidade Federal de Pernambuco,
50.670-901, Recife, PE, Brazil
Corresponding author: Santana Oliveira, Idjane
(idjaneoliveira@gmail.com)
Current Opinion in Food Science 2019, 29:64–79
This review comes from a themed issue on Food mycology
Edited by Marina Copetti
https://doi.org/10.1016/j.cofs.2019.08.004
2214-7993/ã 2019 Elsevier Ltd. All rights reserved.
Introduction
The scientific literature estimates the existence of about
1 million different fungal species [1]. Its well-known
applicability in human and animal life places the fungal
kingdom to be constantly exploited industrially through
Current Opinion in Food Science 2019, 29:64–79
the production of chemicals and antibiotics. In addition,
beneficial activities of fungi in food and beverage, espe-
cially in fermentation, increasing the nutritional and
commercial value [2–4]. In contrast, some studies esti-
mate that over 400 fungi species present pathogenicity to
humans with well-defined action mechanisms, which
fungal infections are basically divided into two groups:
endemic (caused by ‘true pathogenic’ fungi species) and
opportunistic (affecting immunocompromised patients)
pathogens [5,6].
Toxins (from the Greek toxikon) are poisonous substances
produced by several microorganisms, plant and animal
species, mainly found in the form of peptides or proteins
leading to severe disease after interacting with enzymes
and cellular receptors. Mycotoxins are presented as small
secondary metabolites produced by some fungi species
(MW 700), becoming one of the main causes in food
poisoning, reaching both human and animal’s health after
the ingestion of contaminated products [7]. Mycotoxins
are extremely resistive in nature, affecting a variety of
crops before and after harvest. Mycotoxins can be found
in beverages such as beer and wine after the use of
contaminated materials such as the juice of selected
fruits, grapes, and cereals in drink production. In addition,
due to the initial contamination of livestock animals feed,
these hazardous toxins can also reach humans in contam-
inated milk, eggs and meat [8,9].
Three main fungal genera stand out in mycotoxin pro-
duction in foodstuff: Aspergillus, Fusarium, and Penicillium
[10]. Some mycotoxins and their fungal species are listed
in Table 1. Considering the economic impact and poten-
tial risk to human health, a distinct group of mycotoxins
are highlighted, which include aflatoxins, ochratoxin A
(OTA), zearalenone (ZEN), fumonisins, ergopeptine
alkaloids and trichothecenes such as deoxynivalenol
(DON), T-2 and HT-2 toxins [11]. However, other haz-
ardous mycotoxins should be included as a threat due to
their unusual frequency in food and feed contamination.
In this sense, the list expands with patulin (PAT),
cyclopiazonic acid, citrinin, gliotoxin, sterigmatocystin,
penitrems, and others [12–14].
Adverse effects take place in the host cells after exposure
to mycotoxins, leading to oxidative stress, DNA damage,
and cell death. According to the type of mycotoxin,
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 Biosensor for detection of fungi and mycotoxins in food Santana Oliveira et al. 65

Table?1

Summary of the main mycotoxins and corresponding producing species

Mycotoxin Fungal species

Aflatoxins B1, B2, G1, G2, M1 Aspergillus section Flavi
Ochratoxin A Aspergillus section Circumdati Aspergillus section Nigri Penicillium verrucossum
Zearalenone Fusarium graminearum F. culmorum F. cerealis
Deoxynivalenol (type B trichothecenes) Fusarium graminearum Fusarium culmorum
Fumonisin Fusarium section Liseola Aspergillus section Nigri
Ergot alkaloids Claviceps genus Acremonium coenophialum
T-2 toxin (type A trichothecenes) Fusarium sprotrichioides F. poae F. acuminatum
HT-2 toxin (type A trichothecenes) Fusarium sprotrichioides F. poae F. langsethiae
Citrinin Aspergillus niger Penicillium citrinum P. verrucosum
Cyclopiazonic acid Aspergillus flavus Penicillium commune P. camamberti
Gliotoxin Aspergillus fumigatus
Patulin Penicillium expansum P. griseofulvum P. carneum
Roquefortine Penicillium roqueforti
Diacetoxyscirpenol (type A trichothecenes) Fusarium sporotrichiodes
Penitrems Penicillium crustosum
Sterigmatocystin Aspergillus versicolor A. flavus

similar or specific pathways (triggering of the ribotoxic
stress response and protein synthesis inhibition caused by
T-2 toxin and OTA, respectively) to reach cellular tox-
icity can be observed [15]. According to the mycotoxin
structure, the biological response ranges from headache,
diarrhea, and vomiting, to more severe cases with a
mutagenic, nephrotoxic, carcinogenic and immunosup-
pressive character, resulting in the host death [16,17].
The International Agency for Research on Cancer (IARC)
classified mycotoxins based on their carcinogenicity. The
IARC classifies mycotoxins in three main groups, as
follows: aflatoxins (AFB1, AFB2, AFG1, and AFG2) were
classified as carcinogenic to humans (Group 1), followed by
AFM1, OTA, fumonisin B1/B2, fusarin C, and sterigma-
tocystin as possibly carcinogenic to humans (Group 2B) and
finally citrinin, cyclochlorotine, DON, fusarenone X,
nivalenol, PAT, T-2 toxin, and ZEN as not classifiable
as to its carcinogenicity to humans (Group 3) [12,18,19].
World Health Organization (WHO), EU Scientific Com-
mittee for Food (SCF) and others developed specific
regulations and recommendations in order to determine
the maximum levels of mycotoxins in foodstuff [20].
Table 2 shows the maximum levels of aflatoxins, OTA,
PAT, DEN, ZEN, fumonisins, T-2 and HT-2 in food and
feed in mg/kg [21–24].
The early detection, identification, and quantification of
mycotoxins have become essential to avoid the economic
losses, which involves agricultural commodities, product
recalls, adverse publicity for the feed and food industry,
death of animals in livestock and acute and chronic effects
on human health, resulting in high-cost treatments.
Mycotoxins are usually found at trace levels, such as parts
per billion (ppb) and nanograms, which makes detection
difficult [25,26]. In this sense, different quantification
techniques have been developed, and in fact, the demand
for fast and economical techniques for the determination
of mycotoxins in quality food control has increased
significantly.
Rapid screening assays arise as a simple and effective
choice among qualitative tests for monitoring mycotoxins
in crops, food, and feed. The main advantages of these
tests also rely on the simplicity for analyte detection,
sample preparation, and low cost of analysis [27]. Despite
their good specificity, the lack in reproducibility, sensi-
tivity, and reliability are usual limitations, since often the
false-positive results are still reported and correlated to
cross-reactions between antibody and fungal metabolites.
Results with a low recurrence (i.e. <5%) are acceptable.
However, even using well-established screening proce-
dures, samples are usually submitted to traditional tests
before acceptance or rejection of a batch [28].
Rapid diagnostic methods for mycotoxin detection are
mainly based in immunochemical assays, whose dipstick,
flow-through membranes and lateral-flow (LFD) devices
stand out as prime examples. With application mainly
exploited outside the laboratory environment, dipstick
(working similarly to ELISA) and flow-through immu-
noassays present qualitative and semi-quantitative results
close to the cutoff level, thus presenting a low commercial
performance [28,29]. LFD is a fast immunochromato-
graphic test that works in a competitive method, using
a labeled antibody as a signal reagent [30]. Innovative
labels have been developed aiming a signal amplification
in LFD by means of the application of nanoparticles such
as quantum dots (QT) [31], gold nanoparticles (AuNPs)
[32], magnetic nanoparticles (Fe3O4) [33], carbon nano-
particles (CNPs) [34], among others.
Sensitive and accurate analytical methods are required
since the levels of mycotoxins found in food, crops and
their byproducts are extremely low (usually ppb concen-
trations) (Figure 1). Performance parameters of these

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66 Food mycology

Table?2

The maximum permitted levels of mycotoxins in foodstuff (Commission Regulation No 1881/2006 and Commission Recommendation
2013/165/EU)

Mycotoxins Maximum levels mg Kg 1

(ppb)
B1 Sum of B1, B2, G1, and G2 M1
Aflatoxin
» Groundnuts and other oilseeds to be subjected to sorting, or other physical 8.0 15.0 x
treatment, before human consumption or use as an ingredient in foodstuffs
(except groundnuts and other oilseeds for crushing for oil production;
» Almonds, pistachios and apricot kernels to be subjected to sorting, or other 12.0 15.0 x
physical treatment, before human consumption or use as an ingredient in
foodstuffs;
» Hazelnuts and Brazil nuts, to be subjected to sorting, or other physical 8.0 15.0 x
treatment, before human consumption or use as an ingredient in foodstuffs;
» Tree nuts, other than the tree nuts to be subjected to sorting, or other physical 5.0 10.0 x
treatment, before human consumption or use as na ingredient in foodstuffs;
» Groundnuts (peanuts) and other oilseeds and processed products thereof, 2.0 4.0 x
intended for direct human consumption or use as an ingredient in foodstuffs,
except crude vegetable oilsand refined vegetable oils;
» Almonds, pistachios, and apricot kernels intended for direct human 8.0 10.0 x
consumption or use as an ingredient in foodstuffs;
» Hazelnuts and Brazil nuts, intended for direct human consumption or use as an 5.0 10.0 x
ingredient in foodstuffs;
» Dried fruit to be subjected to sorting, or other physical treatment, before human 5.0 10.0 x
consumption or use as an ingredient in foodstuffs;
» Dried fruit and processed products thereof, intended for direct human 2.0 4.0 x
consumption or use as an ingredient in foodstuffs;
» All cereals and all products derived from cereals, including processed cereal 2.0 4.0 x
products;
» Maize to be subjected to sorting or other physical treatment before human 5.0 10.0 x
consumption or use as an ingredient in foodstuffs;
» Raw milk, heat-treated milk, and milk for the manufacture of milk-based x x 0.050
products;
» Spices (dried fruits thereof, whole or ground, including chilies, chili powder, 5.0 10.0 x
cayenne and paprika, white and black pepper, nutmeg, ginger, turmeric);
» Infant formulae and follow-on formulae, including infant milk and follow-on milk; x x 0.025
» Dietary foods for special medical purposes intended specifically for infants 0.10 x 0.025
Ochratoxin A
» Unprocessed cereals; 5.0
» Products derived from unprocessed cereals, including processed cereal 3.0
products and cereals intended for direct human consumption;
» Dried vine fruit (currants, raisins and sultanas); 10.0
» Roasted coffee beans and ground roasted coffee, excluding soluble coffee; 5.0
» Soluble coffee (instant coffee); 10.0
» Wine (including sparkling wine,aromatized wine and wine-product cocktails, 2.0
excluding liqueur wine) grape juice and nectar for direct human consumption;
» Processed cereal-based foods and baby foods for infants and young children; 0.50
» Dietary foods for special medical purposes intended specifically for infant 0.50
Patulin
» Fruit juices, concentrated fruit juices as reconstituted and fruit nectar; 50
» Spirit drinks, cider, and other fermented drinks derived from apples or 50
containing apple juice;
» Solid apple products, including apple compote, apple pure intended for direct 25
consumption;
» Apple juice and solid apple products for infants and young children; 10.0
» Baby foods other than processed cereal-based foods for infants and young 10.0
children
Deoxynivalenol
» Unprocessed cereals other than durum wheat, oats, and maize; 1250.0
» Unprocessed durum wheat and oats; 1750.0
» Unprocessed maize, with the exception of unprocessed maize intended to be 1750.0
processed by wet milling;
» Cereals intended for direct human consumption, cereal flour, bran and germ as 750
end product marketed for direct human consumption;
» Pasta (dry); 750

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 Biosensor for detection of fungi and mycotoxins in food Santana Oliveira et al. 67

Table?2 (Continued )
Mycotoxins Maximum levels mg Kg 1
(ppb)
B1 Sum of B1, B2, G1, and G2 M1
» Bread (including small bakery wares), pastries, biscuits, cereal snacks, and 500
breakfast cereals;
» Processed cereal-based foods and baby foods for infants and young children; 200
» Milling fractions of maize with particle size > 500?microns; 750
» Milling fractions of maize with particle size  500?microns 1,250
Zearalenone
» Unprocessed cereals other than maize; 100
» Unprocessed maize with the exception of unprocessed maize intended to be 200
processed by wet milling;
» Cereals intended for direct human consumption, cereal flour, bran, and germ; 75
» Refined maize oil; 400
» Bread, pastries, biscuits, cereal snacks and breakfast cereals, excluding 50
maize-snacks and maize-based breakfast cereals
» Maize intended for direct human consumption, maize-based snacks, and 100
maize-based breakfast cereals;
» Processed cereal-based foods (excluding processed maize-based foods) and 20
baby foods for infants and young children;
» Processed maize-based foods for infants and young children; 20
» Milling fractions of maize with particle size > 500?microns; 200
» Milling fractions of maize with particle size  500?microns 300

Mycotoxins Maximum levels mg Kg 1

(ppb)
Sum of B1 and B2
Fumonisins
» Unprocessed maize, with the exception of unprocessed maize intended to be processed by wet milling; 4000.0
» Maize intended for direct human consumption, maize-based foods for direct human consumption; 1000.0
» Maize-based breakfast cereals and maize-based snacks; 800
» Processed maize-based foods and baby foods for infants and young children; 200
» Milling fractions of maize with particle size >500 microns; 1400.0
» Milling fractions of maize with particle size 500 microns 2000.0

Toxin T-1 and toxin HT-2

» Unprocessed cereals
- barley (including malting barley) and maize: 200
- oats (with husk): 1000
- wheat, rye, and other cereals: 100
» Cereal grains for direct human consumption
- oats: 200
- maize: 100
- other cereals: 50
» Cereal products for human consumption
- oat bran and flaked oats: 200
- cereal bran (except oat and milling bran / maize milling products): 100
- other cereal milling products: 50
- breakfast cereals including formed cereal flakes: 75
- bread (including small bakery wares), pastries, biscuits, cereal snacks, pasta: 250
- cereal-based foods for infants and young children: 15
» Cereal products for feed and compound feed
- oat milling products (husks): 200
- other cereals products: 500
- compound feed: 250

methods rely on the limit of detection (LOD) and limit of
quantification (LOQ) to estimate the level of the
detected analyte [35]. Currently, one of the most explored
techniques involve the use of separation/chromatographic
methods such as thin layer chromatography (TLC), cap-
illary electrophoresis (CE), gas chromatography (GC)
linked to mass spectrometry (MS), flame ionization
detector (FID), electron capture detector (ECD) or Four-
ier transform infrared spectroscopy (FTIR); high-perfor-
mance liquid chromatography (HPLC) coupled to ultra-
violet (UV), fluorescence (FLD) and MS detectors,
among others [36,37]. Briefly, the majority of chromatog-
raphy methods work separating mycotoxins within a
carrier such as liquid or gas (mobile phase) through a

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68 Food mycology

Figure 1

Spoilage Moulds

Analytical techniques for evaluation of

Microbiological, spectroscopic ( HIS, NIRS),
imunological (ELISA), genomic (PCR – constitutive gene,
LAMP), proteomic (MALDI TOF MS), sensor (E-nose),
and biosensor (O2, antibody)
contaminated food

Toxigenic Fungi
Microbiological, spectroscopic, immunological (ELISA),
genomic (PCR) – specific gene, LAMP), proteomic
(MALDI TOF MS), and biosensor (DNA, antibody,
aptamer, and others)

Mycotoxins
Chromatography (HPLC), spectroscopy (NMRS),
imunological (ELISA), and biosensor ( antibody,
aptamer, and others)

Current Opinion in Food Science

Analytical techniques for microorganism detection in food.

column (stationary phase) containing adsorbents that will Immunochemical tests are known due to simplicity,
separate the mycotoxin from sample components to be multiple sample reading and low cost. Chromatographic
identified by specific detectors [35]. In addition, another approaches present sensitivity, accuracy and multi-target
widely used method to mycotoxin detection relies on detection. Molecular techniques present a well-known
immunoassays such as enzyme-linked immunosorbent sensitivity. However, these techniques require a time-
analysis (ELISA) which basically explores the signal consuming sample pretreatment consisting of specific
developed by chromogenic substrates after the formation steps (sampling, homogenization, extraction and clean-
of an antigen-antibody complex, leading to a measurable up step to obliterate matrix components), expensive
signal [38]. Competitive assays are generally employed in equipment/reagents, and skilled personnel for operation
ELISA tests due to the size of mycotoxins (lower than [44]. In addition, these methods are not able to detect
1 kDa) as compared to sandwich immunoassay where fungi without pre-treatment and for early monitoring of
multiple antibodies bind to a single toxin [39]. Non- plant diseases in the field [29,45].
destructive methods, such as hyperspectral imaging
(HSI) and near-infrared spectrometry (NIRS), are used Biosensors are an effective tool to amplify the possibility
to evaluate the fungi contamination in food. However, of fungi and mycotoxins identification. Biosensors repre-
these techniques require a considerable amount of spec- sent technology that can be applied to several sectors of
tral pretreatment to establish complex models [40,41]. the food industry from the storage of grains and raw
Molecular approaches such as PCR-based methods are materials, food production/processing, security and pro-
also explored, aiming the identification of mycotoxigenic tection, packaging of food (Figure 2) [46]. They are
fungi species in agricultural commodities and food pro- extremely sensitive and simple to operate, promoting
ducts [42]. Moreover, specific probes can be tested to fast and reproducible analysis, followed by the advantage
determine regulatory genes in metabolic mycotoxin path- of low-cost tools, miniaturization, and development of
ways and their toxigenic potential, once weather and portable devices. Moreover, the world biosensors market
environmental factors in different geographical areas has considerably grown, which was worth about US$
influence directly in mycotoxin production [17,43]. 7.3 billion in 2013 with an estimative to worth US$

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 Biosensor for detection of fungi and mycotoxins in food Santana Oliveira et al. 69
Figure 2
- Food and feed industry
- Agriculture
- Supermarkets
Analyte
- Contaminated Foods
Transducer
- Biosensors
Sensor layer
Scheme of the production/processing, contamination, and identification of the microorganisms in food.
33 billion in 2027, highlighting its applicability in point-
of-care testing and quantification of mycotoxins in
agriculture and foodstuffs [47,48].
Basically, a biosensor comprises a biological sensing ele-
ment that will interact specifically to the analyte, a
transducer able to convert the binding event in a measur-
able signal and an output system [49]. A wide variety of
transducers can be explored for mycotoxin and spoilage,
fungi detection, where optical (surface plasmon reso-
nance – SPR and fluorescence), piezoelectric (quartz
crystal microbalance – QCM), and electrochemical (impe-
dimetric, potentiometric and amperometric) spectrosco-
pies stand out as main biosensing methods [50,51].
Biological materials (e.g. peptides, enzymes, antibodies,
cells, nucleic acids) and bioinspired tools such as
aptamers, molecularly imprinted polymers (MIPs) and
recombinant antibodies present a high focus as sensing
elements on biosensor studies [52,53].
Moreover, biosensors usually seek the application of
nanomaterials as signal amplification probes to improve
the sensitivity of the biosensor. Previous studies
highlighted the use of a wide range of metal nanoparti-
cles, QDs (quantum dots), carbon nanotubes (CNTs) and
nanofibers due to their unique physicochemical proper-
ties, high surface-volume ratio and biocompatibility
[54,55]. These features have become useful in biosensor
development, assisting the functionalization of sensor
platforms for the most diverse biological targets, such
as mycotoxins present in foodstuff and beverages
[51,56,57]. The present review summarizes the current
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Signal processor Monitoring system
Current Opinion in Food Science
knowledge on early detection of spoilage, toxigenic fungi
and mycotoxins in foods using biosensor technology.
Biosensing strategies for the detection of
mycotoxins in feed and food
The development of new biodevices is extremely
required for spoilage fungi detection with fast, sensitive,
and non-destructible characteristics to monitor early dete-
rioration fungi in the industry. In this sense, this section
and Table 3 summarizes recent trends for the detection
and quantification of mycotoxin.
Electrochemical biosensors for mycotoxins
Impedimetric sensors
Electrochemical impedance spectroscopy (EIS) tech-
nique arises as a useful tool for mycotoxins identification
and monitoring the changes that occur in the interface
originated between an electrode surface modified by a
nanostructured platform in contact with a redox probe
(e.g. Fe(CN)63 /4 , Ru(NH3)63+) [58]. A three-electrode
system is conventionally used to electroanalytical appli-
cations, as follow: 1) working electrode (WE), substrate of
biosensor platform assembly where reactions and electron
transfer occur; 2) reference electrode (RE), usually
Ag/AgCl, which promotes a stable and reproducible
potential; and, 3) counter electrode (CE), providing the
electric current through the electrochemical solution to
WE [59,60].
EIS present as main parameter the charge transfer resis-
tance (RCT) values, whose variation is closely related to
the biding event (e.g. mycotoxin immobilization and
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70 Food mycology

Table 3

Examples of biosensors developed for detection and quantification of mycotoxins

Mycotoxin Recognition element Transducer/technique Matrix LOD Reference

AFB1 Aptamer Flurorescence (FRET) Maize and wheat 0.0004 mg mL 1
[132]
AFB1 Antibody Impedimetric (CV/EIS) Corn flour 0.79 pg g 1 [62]
AFB1 Organic framework Piezoelectric (QCM) Peanut, pistachio, 2.8 pg mL 1 [95]
composite rice, and wheat
AFB1 Antibody Impedimetric (EIS) Corn 0.05 ng mL 1 [63]
AFB1 Antibody Piezoelectric (QCM) x 1.625 ng mL 1 [98]
AFB1 Antibody Piezoelectric (QCM) Peanut 0.83 ng kg 1 [133]
AFB1 Antibody Potentiometric (DPV) Corn powder 3.5 pg mL 1 [71]
AFB1 AFB2 Aptamer Optical (SPR) Vinegar 0.19 ng mL 1 [86]
Citrinin Antibody Flurorescence (FRET) Human serum 0.1 pM [92]
Cyclopiazonic acid Antibody Optical (SPR) Maize and cheese 0.29 ng mL 1 [134]
DON ZEN T-2 toxin Antibody Optical (SPR) Wheat 15 mg kg 1 24 mg kg 1
[85]
12 mg kg 1
Fumonisin B1 Aptamer Impedimetric (EIS) Maize 2 pM [135]
HT-2 toxin T-2 toxin AFM1 Antibody Amperometric Human urine 0.4 ng mL 1 1 ng mL 1
[136]
0.3 ng mL 1
OTA Aptamer Impedimetric (EIS) Grape and 0.030 ng mL 1 [137]
commodities
1
OTA Aptamer Amperometric (CV) Red wine 0.23 pg mL [138]
OTA Antibody Piezoelectric (QCM) Red wine 0.16 ng mL 1 [139]
OTA Antibody Optical (SPR) Coffee 3.8 ng mL 1 [140]
OTA Aptamer Impedimetric (EIS) Fruit juice 5.2 fg mL 1 [141]
OTA Black phosphorene Potentiometric (DPV) Grape juice and 0.18 mg mL 1 [142]
red wine
1
OTA Antibody Piezoelectric (QCM) x 17.2 ng mL [99]
1 1
OTA AFM1 Antibody Potentiometric (CV) Red wine and milk 0.15 ng mL 3.04 ng mL [70]
1
PAT Aptamer Potentiometric (EIS/DPV) Juice 0.27 pg mL [143]
PAT Aptamer Impedimetric (EIS) Apple juice 2.8 ng L 1 [66]
Sterigmatocystin Soybean peroxidase Amperometric Corn 2.3 nM L 1 [75]
enzyme
T-2 toxin Aptamer Flurorescence (FRET) Maize and wheat 0.93 pg mL 1 [93]
T-2 toxin Antibody Optical (SPR) Wheat 1.2 ng mL 1 [144]
ZEN Aptamer Amperometric (CV/DPV) Maize 0.17 pg mL 1 [145]
ZEN Antibody Amperometric (CV/DPV) Corn and corn 1.5 pg mL 1 [76]
products
ZEN Antibody Impedimetric (capacitive) x 1.9 pg mL 1 [146]
1
ZEN Aptamer Potentiometric (CV/DPV) Maize 0.105 pg mL [72]

antibody-antigen reaction) and proportional to the ana-
lyte/target concentration [61,62]. Yagati et al. [63] devel-
oped a label-free impedimetric immunosensor composed
by polyaniline (PANI) nanofibers coated with AuNPs in
indium tin oxide (ITO) disk electrodes to improve the
efficiency on the detection of AFB1 in spiked corn,
enabling the loading of multiple capture antibodies sites.
Yagati et al. [63] monitored the changes in impedance
magnitude (|Z|) by EIS, detecting AFB1 with high
sensitivity, linear range of 0.1–100 ng mL 1 and LOD
of 0.05 ng mL 1.
Aptamers are becoming a growing alternative in myco-
toxin detection [64]. Aptamers structure is composed of
single-stranded oligonucleotides (RNA or DNA) can bind
with high specificity to a wide range of targets, including
mycotoxins and toxigenic fungi. Moreover, aptamer pro-
duction is cheaper than that observed in antibodies and
can be easily labeled in biosensing platforms [65]. A
recent study showed the use of aptamers to the label-
free detection of PAT in apple juice accessed by EIS [66].
The assembling of the anti-PAT-aptamer onto the sur-
face of the screen-printed gold electrodes (AuSPE)
enabled the detection of PAT with high sensitivity
(LOD and LOQ of 2.8 ng L 1 and 4.0 ng L 1, respec-
tively) and selectivity against ZEN, OTA, ochratoxin B,
and AFB1.
Recently, an impedimetric sensor for AFM1 detection
based on aptamers has been reported by Istamboulié et al.
[67]. In this work, hexaethyleneglycol-modified oligonu-
cleotides (seven-base pair sequences) of the anti-AFM1
aptamer were immobilized on the diazotized screen car-
bon electrode by a carbodiimide coupling reaction. A
dynamic range of 2–150 ng L 1 was obtained with a
LOD of 1.15 ng L 1. Krittayavathananon et al. [68] devel-
oped an impedimetric sensor for AFB1 detection in milk
using a single-stranded DNA covalently attached to a
functionalized reduced graphene oxide aerogel deposited
on a glassy carbon rotatory electrode (LOD of 0.04 ng

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 Biosensor for detection of fungi and mycotoxins in food Santana Oliveira et al. 71
mL 1). Of note, EIS biosensors do not require an enzyme
conjugate to antibodies or mycotoxins to obtain an elec-
trochemical signal [69].
Potentiometric sensors
The potentiometric biosensors address the use of ion-
selective electrodes to achieve analytical data. The poten-
tiometric method requires two (WE and RE) or three
(WE, RE and CE) electrode systems, where the recogni-
tion event is evaluated through the variations in the
circuit potential between the WE and RE [49]. Different
voltammetric techniques such as cyclic voltammetry
(CV), differential pulse voltammetry (DPV) and
square-wave voltammetry (SWV) have been used in
mycotoxin measurements.
Karczmarczyk et al. [70] reported an inhibition-based
assay in AuSPE aiming the detection of OTA and
AFM1 in red wine and milk, respectively. The sensing
platform consisted by a self-assembled monolayer (SAM)
of 3-mercaptopropionic acid and BSA-mycotoxin conju-
gates enabled the sensitive detection of the toxins
through CV and DPV with a LOD of 15 ng mL 1 to
OTA and 37 pg mL 1 to AFM1. In a study conducted
by Zhang et al. [71], a glassy carbon electrode was modi-
fied with a single/walled carbon nanotube (SWCNTs)/
chitosan nanocomposite as immunosensor for AFB1
detection in corn. Electrochemical measurements includ-
ing DPV, CV, and SWV showed a linear range of 0.001–
100 ng mL 1 and LOD of 3.5 pg mL 1. Recently, a vol-
tammetric aptasensor was developed containing Fe3O4
and hollow cubic platinum/gold nanoframes to enhance
the sensitive detection of ZEN in maize samples [72].
The capture probe, a DNA S2-aptamer immobilized
on the sensing platform, was able to detect ZEN with
good reproducibility and stability, with a LOD of
0.105 pg mL 1.
Amperometric sensors
Similar to potentiometric studies, a typical amperometric
sensor also works with a two or three-electrode system.
The identification of an analyte by an amperometric
transducer is carried out through the measurement of
the resulting current generated after the reduction and
oxidation of electroactive species immobilized on the WE
surface after setting an appropriate potential [59,73]. The
WE are usually made of inert metals (Pt, Au) or carbon
(graphite, vitreous carbon). However, the main disadvan-
tage is the regeneration between measurements. Cur-
rently, disposable printed electrodes have been an out-
standing alternative due to low cost and scale production
[69,74].
Dı́az Nieto et al. [75] demonstrated a useful approach for
mycotoxin detection through biosensor technologies
relies on enzyme-modified electrodes. Soybean peroxi-
dase enzyme was chosen as a biological sensing element
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due to its stability in a wide pH range and thermal
stability for the determination of sterigmatocystin in corn
samples. Amperometric results showed a LOD of
2.3  10 9 mol L 1 and good agreement with HPLC mea-
surements. Another study has focused on the use of
monoclonal antibodies for ZEN detection adopting elec-
trodeposited multi-walled carbon nanotubes (MWCNTs)
and gold–platinum (AuPt) nanoparticles as signal-
enhancing agents in the sensor platform [76]. This amper-
ometric immunosensor was able to detect ZEN in corn
flour and corn-based baby food using CV and DPV with a
linear range of 0.005–50 ng mL 1 and LOD of
1.5 pg mL 1. Tria et al. [77] developed an innovative
device with the capability to detect OTA by simultaneous
amperometric and luminescent evaluation. Tria et al. [77]
explored the use of the synthetic peptide NFO4 with
high binding affinity to OTA based on a microfabricated
cell-on-a-chip transducer with a concentration range of
1–10 mg L 1.
In addition, screen-printed electrodes can be used asso-
ciated with multichannel electrochemical systems for
analysis of multi-AFB1. Piermarini et al. [78] developed
an immunosensor array based on intermittent pulse
amperometry for multi-AFB1 detection in corn samples.
In contrast, magnetized screen-printed electrodes have
been used as electrochemical transducers. Piermarini’s
group developed a device for AFB1 detection based on
indirect competitive ELISA using magnetic beads as
immobilization support and magnetized screen-printed
electrodes [79].
Previous works demonstrate that the enzyme acetylcho-
linesterase (AChE) is inhibited by aflatoxins. Aflatoxin
oxidase incorporated in sol–gel has been used for the
development of AFB1 amperometric biosensor on elec-
trodes of Pt electrode modified by multiple-walled carbon
nanotubes (MWCNTs) with a LOD of 1.6 nmol L 1
(0.5 ng mL 1) [80]. In addition, an enzymatic biosensor
for the evaluation of aflatoxin B1 (AFB1) in olive oil was
developed [81]. The immunosensor and label-free EIS
sensors follow the changes in resistance to electron trans-
fer at the surface of the working electrode resulting from
the complexation of mycotoxins in the range of 1–20 ng
mL 1 with detection limits of about 0.5 ng mL 1.
Optical biosensors for mycotoxins
Surface plasmon resonance sensors
Optical techniques present useful features such as high
sensitivity, label-free detection, and real-time analysis.
Surface plasmon resonance (SPR) and fluorescence
approaches such as fluorescence resonance energy trans-
fer (FRET) are the most exploited methods in optical
sensors studies [82,83]. In brief, SPR analysis occurs after
assembling of a sensing platform composed by biorecog-
nition agents on the surface of an inert metal (usually gold
thin film) placed on the interface of a glass prism. The
Current Opinion in Food Science 2019, 29:64–79
72 Food mycology
glass prism is illuminated by a laser beam in a predeter-
minate angle, resulting in an evanescent wave that pene-
trates in the gold film. Then, as the evanescent wave
extends, it will interact with free oscillating electrons of
the gold film. A shift of the SPR angle is associated with
the modification of the metal surface that affects the
resonance system [39,83]. SPR shift indicates molecular
interaction/binding event and can be measured compar-
ing two reflection spectra or changes in the intensity of
the reflected light.
Joshi et al. [84] described a competitive inhibition immu-
noassay for DON and OTA in beer using an imaging SPR
(iSPR) sensor. The iSPR sensor was composed of 3-D
carboxymethyl dextran containing specific monoclonal
antibodies (mAbs) against DON and OTA. The iSPR
biosensor chip was reused 450 cycles with a LOD of
17 ng mL 1 and 7 ng mL 1 for DON and OTA, respec-
tively. Hossain et al. [85] developed a study based on
multiplexed iSPR assay composed of AuNPs functional-
ized with secondary antibodies as signal amplification tag
for detection of three different Fusarium toxins (DON,
T-2, and ZEN) in wheat. The antigen-coated biosensor
platform showed excellent reusability and exhibited a
LOD of 15 mg kg 1 for DON, 12 mg kg 1 for T-2 toxin
and 24 mg kg 1 for ZEN.
Recently, Wu et al. [86] reported an aptamer-based SPR
biosensor able to detect two different aflatoxins using the
same aptamer, an unusual phenomenon. The sensing
platform assembled on a CM5 sensor chip was composed
by a thin layer of carboxymethylated dextran covalently
bounded the biotin-modified aptamer using streptavidin
and EDC:NHS coupling agents. The aptasensor detected
both AFB1 and AFB2 with high specificity in vinegar
samples, showing a linear range of 1.5–50 ng mL 1 and
LOD of 0.19 ng mL 1.
Fluorescent sensors
Another powerful class of optical biosensors is found in
fluorescence-based transducers. As can be seen in SPR
sensors, the label-free detection of multiple analytes and
high sensitivity allow their use in academic and medical
purposes as a bioanalytical and diagnostic tool [87]. Fur-
thermore, the association of nanoparticles and fluores-
cence techniques for mycotoxin identification became a
powerful strategy for fluorescence recovery and enhance-
ment of the biosensor sensitivity [88].
Recently, Tan et al. [89] functionalized mesoporous silica
nanoparticles (MSN) with rhodamine 6G as signal fluo-
rescence indicator and specific aptamers as recognition
probes to produce a fluorescence aptasensor for AFB1.
The signal probe is released and a fluorescent signal is
obtained when the aptamer interacts with AFB1. The
sensing platform tested in peanut oil, corn, oatmeal, and
rice showed high specificity to AFB1 against diverse
Current Opinion in Food Science 2019, 29:64–79
mycotoxins and enhanced sensitivity with a LOD of
0.13 ng mL 1.
Bueno et al. [90] developed a fluorescence analyzer based
on a smartphone camera, whose biosensor system consists
basically of a black polymethyl methacrylate and
photometric measurements in the UV range. The device
was able to detect OTA in beer with a linear range of
2–20 mg L 1 and LOD of 2 mg L 1. Bueno et al. [90]
demonstrated how associate innovative technologies to
actual models of health and food quality monitoring.
FRET technology has been extensively used for the
development of fluorescent biosensor based on nonradio-
active energy transferred from an excited donor fluoro-
phore to a nearby acceptor species [91]. Functionalized
nanoparticles play an essential role in FRET analyses as
energy donors. Shojaee Sadi et al. [92] created a FRET-
based immunosensor for citrinin detection in human
serum. The nanobiosensor was developed by assembling
Fe3O4/silica core-shell nanoparticles functionalized with
anti-citrinin antibody and conjugated with BSA/citrinin/
Rho123 fluorophore as acceptor and QDs as energy
donors. FRET-based immunosensor was able to detect
citrinin with a noteworthy LOD of 0.1 pM in a range of
1–6 pM.
Khan et al. [93] used aptamer functionalized silver
nanoclusters assembled on a molybdenum disulfide thin
layer for the development of a selective fluorescent
aptasensor for T-2 toxin detection (LOD of 0.93 pg
mL 1) in maize and wheat samples. The sensing probe
reveals a promising application of the same nanostruc-
tured platform to the detection of other mycotoxins by
changing the aptamer sequence.
Piezoelectric biosensors for mycotoxins detection
Quartz crystal microbalance (QCM)
The potential of QCM-based biosensors has been
explored in the monitoring of toxins and pathogenic fungi
in food and environmental analysis. A QCM transducer is
composed by gold-plated crystal quartz, whose surface is
functionalized with a sensing layer of interest. A molecu-
lar recognition or binding event in the gold electrode
surface leads to a mass change and specific vibrations as an
electric signal is sent through the quartz crystal, inducing
changes in the resonant frequency [94].
In a recent study, a QCM sensor was developed for the
determination of AFB1 [95]. The enhancement of recog-
nition sites and the resulting improvement of sensitivity
in the sensing platform was obtained after the introduc-
tion of MIPs of poly(o-aminothiophenol) and AuNPs
associated with organic frameworks composites. The
proposed sensor was able to detect AFB1 in peanut,
pistachio, rice, and wheat samples with a LOD of
2.8 pg mL 1. The application of MIPs in biosensors for
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 Biosensor for detection of fungi and mycotoxins in food Santana Oliveira et al. 73
mycotoxin detection has become an excellent strategy to
overcome the restrictions of specific antibodies in detect-
ing only one type of target analyte. The main features of
MIPs rely on the ability to mimic recognition elements (e.
g. antibodies, biological receptors) [29,96,97]. Ertekin
et al. [98] developed a novel QCM-based label-free
immunosensor for AFB1 detection using IgA mAb as a
sensing element. The sensor system was composed by a
SAM of 11-mercaptoundecanoic acid (MUA) and IgA
showed a LOD of 1.625 ng mL 1 with a 1.25–10 ng mL 1
detection range. Similarly, Pirinçci et al. [99] developed a
label-free QCM immunosensor obtained with an ami-
nated gold sensor surface with the 10F4 antibody as a
recognition element to OTA. The obtained sensor
expressed a LOD of 17.2 ng mL 1.
Current trends for the detection of spoilage
and toxigenic fungi in food
The presence of spoilage microorganisms in food remains
a threat in public health and food quality directly affect-
ing consumer’s life, which even with modern agricultural
practices in the developed world, millions are affected
with food poisoning [100]. Moreover, the greater impact is
seen in developing countries, where climate and environ-
mental issues aligned to old-fashioned storage procedures
and unregulated markets result in extensive exposure to
these pathogens [101]. The early detection of microor-
ganisms in the whole food chain arises as a crucial step to
reduce economic losses and prevent human and animal
intoxication by mycotoxins (secondary metabolites exten-
sively produced by molds and toxigenic fungi). This
section aims to highlight the current trends in sensing
methods for the detection of mycotoxin-producing micro-
organisms in foodstuffs and agricultural commodities.
A wide range of methods for detecting mycotoxin-pro-
ducing species are available, where the analysis time and
sensitivity play a crucial factor of choice. We can highlight
some methods, as follows: 1) standard microbiological
methods: the first step of procedure relies on microbial
culture, isolation and further identification (e.g. classical
culture-based methods, extremely time-consuming); 2)
proteomic and genomic methods: initial mold peptide/
DNA/RNA extraction from food and further analysis by
matrix-assisted laser desorption/ionization time-of-flight
(MALDI-TOF) and other molecular techniques such as
polymerase chain reaction (PCR), fluorescent in situ
hybridization (FISH), DNA barcoding; and, 3) evaluation
of the physicochemical properties of the metabolites
produced by mycotoxin-producing fungi through elec-
tronic nose (e-nose), and others [42,102].
PCR and its variants such as quantitative PCR (qPCR),
reverse transcription PCR (RT-PCR), PCR denaturing
gradient gel electrophoresis (PCR-DGGE), PCR-ELISA,
and others can be applied for fungi screening in foods,
presenting specific protocols and genes established for
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this purpose [103]. For example, a method to measure
Aspergillus flavus (a potential aflatoxin producing fungi)
biomass in maize based on qPCR assay was recently
described. Mitema et al. [104] demonstrated that A. flavus
b-tub gene was able to detect and quantify A. flavus
biomass in different maize lines found in Africa, placing
the b-tub gene as a potential biomarker for molecular
analyses. Thus, Mitema et al. [104] developed a biocon-
trol strategy for aflatoxin mitigation using an atoxigenic A.
flavus isolate KSM012 against the aflatoxigenic A. flavus
isolate KSM014 after a co-infection of maize. This
approach could be widespread to other mycotoxin-pro-
ducing species.
Furthermore, PCR-DGGE stands out as a promising
molecular tool for identification fungi identification in a
wide range of food samples, since not all cloning techni-
ques are capable of analyzing different types of samples
[105]. In brief, the suspect food sample is processed and
then the extraction of total DNA is performed. Universal
primers are used to the amplification of the microbial
DNA of taxonomic interest to be separated by acrylamide
gel. The resulting fingerprint bands indicate the micro-
bial contaminant in the sample.
Moreover, PCR-DGGE presents the advantage of clon-
ing, sequencing, and potential of terminal-restriction
fragment length polymorphism analysis (T-RFLP) anal-
ysis, becoming useful for the identification of fungal
contaminants [42,106]. Durand et al. [107] reported that
PCR-DGGE can be applied to the screening of coffee
processing to the determination of ochratoxinogenic fungi
species such as Aspergillus niger, Aspergillus carbonarius,
and Aspergillus ochraceus. Durand et al. [107] demonstrated
that DGGE fingerprints presented the evolution of the
fungal microflora of coffee by the extraction and amplifi-
cation of 28S rDNA from the filamentous fungi species.
PCR-ELISA is a PCR variant useful for fungal analyses.
Recently, Omori et al. [108] reported the accurate and
sensitive detection of toxigenic Fusarium verticillioides
(fumonisin producing Fusarium species) in corn samples
by PCR-ELISA. A FUM21 gene was used for identifica-
tion of DNA of Fusarium sp., Aspergillus sp. and Penicillium
sp. isolates. A LOD of 2.5 pg for DNA fragments of F.
verticillioides was obtained. The sensitivity was potentially
increased for Fusarium species as compared to single PCR
analysis.
Luo et al. [109] described the development of the DNA
amplification technique through loop-mediated isother-
mal amplification (LAMP) for detection of three afla-
toxin-producing Aspergillus species (A. flavus, Aspergillus
parasiticus, and Aspergillus nomius). Luo’s group establish a
LOD of 2.4, 7.6, and 20 pg of Aspergillus DNA in artifi-
cially contaminated samples of nuts, peanuts, and green
coffee beans, respectively. Recently, the LAMP method
Current Opinion in Food Science 2019, 29:64–79
74 Food mycology
was optimized for the detection of Fusarium temperatum,
an emerging maize pathogen and fumonisin B1 producer,
which presents a potential risk to human food and animal
feed supply chains [110]. The method was specific and
sensitive detecting less than 10 pg of fungal genomic
DNA in 60 min.
The most widely used and established sensory method-
ology to monitor fungi in food is the electronic-nose (E-
nose). E-nose can be a GC variant system based on a
solid-state sensor that detects volatile compounds [111].
E-nose sensors are sensitive to different volatile com-
pounds and can generate a fingerprint that represents the
flavor/aroma characteristics of the food. Thus, the
unwanted smell of the food can be detected and reflects
the primary metabolism of fungi [112,113]. In summary,
E-nose has been applied for detection of mycotoxin-
producing fungi species in a wide variety of fruits, such
as apples [114], stored oranges [115], strawberry [116] and
peaches [117]. Gu et al. [118] explored the use of E-nose
for the detection of Aspergillus spp. in rice, being able to
differentiate three different Aspergillus species (Aspergillus
candidus, Aspergillus fumigatus, and Aspergillus clavatus).
Shen et al. [119] associated near-infrared spectroscopy
(NIR) to E-nose to develop a fast and sensitive alternative
of detecting A. ochraceus, A. flavus, and A. parasiticus in
peanuts with a LOD of 0.0808 log CFU g 1.
Electrochemical biosensors are useful to toxigenic-fungi
detection. The methods of fungi identification are mainly
developed to evaluate industrialized products of eco-
nomic importance. Of note, wine is one of the beverages
most studied in the development of biosensors for moni-
toring of fungal contamination. Tubı́a et al. [120] created
an impedimetric immunosensor to detect Brettanomyces
spp. (the main deteriorating yeast of wine). Conventional
commercial methods for Brettanomyces spp. require a long
period of culture and occurs contamination with filamen-
tous fungi, as well as false-positive results. Different
species of wine spoilage yeasts, such as Brettanomyces
bruxellensis, Pichia guillermondi, and Debaromyces hansenii
were evaluated by using an impedimetric immunosensor.
A high impedance variation was obtained when exposed
to yeast species than that observed for label-free detec-
tion of the same analytes. Recently, Villalonga et al. [121]
developed two amperometric biosensors for rapid and
sensitive detection/quantification of B. bruxellensis and
total yeasts in red wine. The sensing platform was based
on superparamagnetic Fe3O4@SiO2 labeled with Conca-
navalin A to yeast detection presenting a LOD of 5 CFU
mL 1. An anti-Brett polyclonal antibody to detection B.
bruxellensis was also used with a LOD of 8 CFU mL 1 in a
wide range of 10–106 CFU mL 1.
Mateo et al. [122] explored a solid-state voltammetric
strategy to create an electrochemical fingerprint model
to detect and differentiate mycotoxin-producing
Current Opinion in Food Science 2019, 29:64–79
Aspergillus and Fusarium species. The obtained SWV peak
potential data were able to precisely discriminate
between genera and species, in a range of 20 different
species. Sedighi-Khavidak et al. [123] were able to detect
A. flavus (potential aflatoxins and sterigmatocystin pro-
ducing Aspergillus species) in pistachio samples combin-
ing CV and EIS analysis associated to electrodeposited
AuNPs and an ssDNA probe of aflD gene onto a GCE.
Sedighi-Khavidak et al. [123] obtained a massive LOD of
0.55 nM and a linear range from 1 nm to 10 mM to the aflD
gene. Silva et al. [124] described the first electrochemical
biosensor for detection of Penicillium sclerotigenum in yam.
The impedimetric genosensor was used to detect fungal
genomic DNA in samples of pure cultures and yams. The
developed sensor was based on a self-assembled mono-
layer of a magnetite/polyallylamine hydrochloride com-
posite as an anchoring layer for DNA hybridization and
used EIS to evaluate and quantify the hybridization
degree. The sensitivity of the sensing platform is notice-
able (LOD of 24 pg mL 1) aligned to specific and repro-
ducible results for the detection of fungi in foods.
Recently, Luna-Moreno et al. [125] developed an immu-
nosensor based on SPR detection for Pseudocercospora
fijiensis, which causes Sigatoka black banana disease, in
banana leaf extracts. The immunosensor detected a pro-
tein (glycosylphosphatidylinositol – GPI) extracted from
the cell wall of P. fijiensis. The gold-coated SPR chips
were functionalized for direct immobilization of antibo-
dies against the HF1 protein (GPI protein of P. fijiensis).
The optical response was concentration dependence to
HF1 and monitored in real-time by SPR (LOD of
2.1 ng mL 1).
An innovative proposal was reported by Zang et al. [126]
to monitor grain spoilage fungi based on O2 biosensor to
detect catalase activity. Zang’s group obtained a high
correlation (>0.99) between catalase activity and spore
counts for four fungal species analyzed (Aspergillus glaucus,
Aspergillus candidum, A. flavus, and Penicillium cyclopium) in
stored grain samples (wheat, rice, and corn). Detection of
catalase activity was performed in 10 min, while culture
and spore counting require five days. The proposed
sensor system is highly sensitive and fast as compared
to conventional monitoring methods.
Matrix-assisted laser desorption/ionization time-of-flight
mass spectrometry (MALDI-TOF MS) has been
highlighted as an attractive and effective tool in monitor-
ing food contaminants, such as spoilage fungi [127].
MALDI-TOF MS is a fast and accurate method in
detecting various fungal groups, especially Aspergillus,
Fusarium and Penicillium spp., well-known mycotoxin-
producing filamentous fungi [128].
MALDI-TOF MS is based on the detection of proteins
through an N2 laser emission to a UV-absorbing matrix
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 Biosensor for detection of fungi and mycotoxins in food Santana Oliveira et al. 75
that covers the suspect sample. The reaction leads to the
formation of gas-phase analyte ions to be calculated mass-
to-charge ratio (m/z) values, leading to the formation of a
unique fingerprint spectrum (specific for each microor-
ganism) [129]. Silva et al. [130] reported the identification
of thirty-five different isolates of Aspergillus section flavi
strains in Brazilian food commodities such as coffee,
Brazil nuts, peanuts, maize, rice, and cereal snack bars.
The spectral results showed that was possible to create a
proteomic profile of aflatoxins-producing Aspergillus spe-
cies. Jeyakumar et al. [131] recently described the deter-
mination of eight different mycotoxins produced by
Fusarium spp. (fumonisin B1 and B2, DEN, nivalenol,
3-acetyl-deoxynivalenol, 15-acetyl-deoxynivalenol,
ZEN, and AFG1) based on HPLC, liquid chromatogra-
phy-electrospray ionization-tandem mass spectroscopy
(LC-ESI-MS/MS), and MALDI-TOF MS. The analysis
was carried out in sugarcane samples infected by Fusar-
ium species (F. verticillioides, F. proliferatum and F. oxy-
sporum). Thus, complementary techniques are promising
in mycotoxin and infectious fungi detection.
Conclusions
Conventional laboratory techniques (e.g. chromatography
and mass spectrometers) remain the first-choice methods
for detecting mycotoxin due to their feasibility, sensitiv-
ity and a wide detecting analyte list. New methods for
food and feed analysis should be explored to detect toxins
and human pathogens at very low levels. The convenient
and straightforward characteristics of innovative biosen-
sors and its plentiful biorecognition methods with a fast,
low-cost, sensitive and selective analysis resulted in their
emerging use in mycotoxin detection for food industry
environmental monitoring aligned to point-of-care test-
ing. Moreover, autonomous operation and simple assem-
bly and detection protocols stand out as the main param-
eter for the continuous growth of the biosensing area.
Developing countries are the major targets of mycotoxins,
spoilage, and toxigenic fungi in agriculture commodities,
foodstuff and human intoxication. Thus, the biosensor
technology is extremely useful as an auxiliary method to
conventional techniques.
Conflict of interest statement
Nothing declared.
Acknowledgements
M.D.L. Oliveira and C.A.S. Andrade would like to thank CNPq (grant
314894/2018-7 and 314756/2018-3).
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