Fungal screening and aflatoxin production by Aspergillus section Flavi isolated from pre-harvest maize ears grown in two Argentine regions

2017 ◽  
Vol 92 ◽  
pp. 41-48 ◽  
Author(s):  
Boris X. Camiletti ◽  
Ada K. Torrico ◽  
M. Fernanda Maurino ◽  
Diego Cristos ◽  
Carina Magnoli ◽  
...  
2018 ◽  
Vol 81 (9) ◽  
pp. 1508-1518 ◽  
Author(s):  
PAUL W. KACHAPULULA ◽  
JULIET AKELLO ◽  
RANAJIT BANDYOPADHYAY ◽  
PETER J. COTTY

ABSTRACT Dried insects and fish are important sources of income and dietary protein in Zambia. Some aflatoxin-producing fungi are entomopathogenic and also colonize insects and fish after harvest and processing. Aflatoxins are carcinogenic, immune-suppressing mycotoxins that are frequent food contaminants worldwide. Several species within Aspergillus section Flavi have been implicated as causal agents of aflatoxin contamination of crops in Africa. However, aflatoxin producers associated with dried fish and edible insects in Zambia remain unknown, and aflatoxin concentrations in these foods have been inadequately evaluated. The current study sought to address these data gaps to assess potential human vulnerability through the dried fish and edible insect routes of aflatoxin exposure. Caterpillars (n = 97), termites (n = 4), and dried fish (n = 66) sampled in 2016 and 2017 were assayed for aflatoxin by using lateral flow immunochromatography. Average aflatoxin concentrations exceeded regulatory limits for Zambia (10 μg/kg) in the moth Gynanisa maja (11 μg/kg), the moth Gonimbrasia zambesina (Walker) (12 μg/kg), and the termite Macrotermes falciger (Gerstacker) (24 μg/kg). When samples were subjected to simulated poor storage, aflatoxins increased (P < 0.001) to unsafe levels in caterpillars (mean, 4,800 μg/kg) and fish (Oreochromis) (mean, 23 μg/kg). The L strain morphotype of A. flavus was the most common aflatoxin producer on dried fish (88% of Aspergillus section Flavi), termites (68%), and caterpillars (61%), with the exception of Gynanisa maja, for which A. parasiticus was the most common (44%). Dried fish and insects supported growth (mean, 1.3 × 109 CFU/g) and aflatoxin production (mean, 63,620 μg/kg) by previously characterized toxigenic Aspergillus section Flavi species, although the extent of growth and aflatoxigenicity depended on specific fungus-host combinations. The current study shows the need for proper storage and testing of dried insects and fish before consumption as measures to mitigate human exposure to aflatoxins through consumption in Zambia.


2011 ◽  
Vol 4 (1) ◽  
pp. 69-77 ◽  
Author(s):  
A. Abdel-Hadi ◽  
D. Carter ◽  
N. Magan

A wide range of Aspergillus section Flavi strains were isolated from Egyptian peanut samples. Eighteen of these strains were compared with two type strains (Aspergillus flavus SRRC G1907 and Aspergillus parasiticus 2747) for aflatoxin production based on (a) qualitative fluorescence using a coconut cream agar medium (CAM), and (b) aflatoxin production on a conducive Yeast Extract-Sucrose (YES) medium using HPLC. These results were validated by using molecular approaches (the structural genes, aflD (nor-1), aflM (ver-1) and aflP (omt A) and the regulatory gene aflR) to discriminate between aflatoxigenic and non-aflatoxigenic strains of the Aspergillus section Flavi group in vitro and on peanut seeds. Overall, 13/18 strains producing aflatoxins B1 and B2 in the range 1.27-213.35 µg/g medium were identified. In addition, 5 non-aflatoxin producing strains were found. The expression of these four genes was assessed using PCR and RT-PCR. PCR showed that all strains contained the four aflatoxin genes examined, regardless of expression profiles. Our results also showed that aflD expression is a reliable marker to discriminate between aflatoxin and non-aflatoxin producers. Interestingly, when an aflatoxin producing strain and three non-aflatoxigenic strains were subsequently grown on peanuts at 0.95 water activity, two of the non-producers were able to initiate aflatoxin biosynthesis. This suggests that growth of strains on the natural food matrix is important for confirming aflatoxigenic production potential.


Author(s):  
Hamidou Compaoré ◽  
Serge Samandoulougou ◽  
Clarisse S. Compaoré ◽  
Alima Bambara ◽  
Hissein Ratongué ◽  
...  

A total of four samples of rice intended for infant flour production in Ouagadougou were received at the Physico-chemistry laboratory of Food Technology Department (DTA) for quality control. The latter were also tested for Aspergillus section Flavi presence and analyzed for aflatoxins B1, B2, G1 and G2 content using high performance liquid chromatography (HPLC). Among the twenty (20) strains of mold isolated from these samples, three Aspergillus section Flavi were obtained and cultivated in “Aspergillus flavus and parasiticus Agar (AFPA)” to ascertain if they belong to Aspergillus flavus or Aspergillus parasiticus species. The qualitative ability of aflatoxin production was also performed by fluorescence emission under ultra violet light at 365 nm after four days of incubation at 30 °C on Coconut Agar Medium (CAM). Statistical analysis results showed that 75% of samples were contaminated with total aflatoxins (AFs) with contents ranging from 0.54 ± 0.06 to 2.40 ± 0.07 µg/Kg. Aflatoxin B1 (AFB1) and aflatoxin B2 (AFB2) were detected in two contaminated samples. AFB1 had the highest concentration as compared with other aflatoxins. A significant level of contamination (p< 0.0001) was observed in sample R441 compared to other sample types.


Author(s):  
I. Lavkor

Totally, 50 Aspergillus section Flavi were identified isolates having aflatoxin biosynthesis genes on peanut by molecular method and aflatoxin production. Primer pair (IGS-F/R) recognized the aflatoxin biosynthesis gene (aflJ-aflR) targeting the intergenic region (IGS) on DNA was amplified by polymerase chain reactions (PCR). The PCR product were restricted by BglII enzyme within Restriction Fragment Length Polymorphism (RFLP) and obtained from 33 (66%) Aspergillus flavus was cleaved into three band sizes of 362, 210 and 102 bp. However, BglII enzyme generated two band sizes of 363 and 311 bp for 17 (34%) Aspergillus parasiticus. An investigation examined DNA sequence data to characterize these isolates and describe the species. Phylogenetic analysis showed that A. flavus and A. parasiticus have been identified in different groups. All the A. flavus and A. parasiticus isolates produced aflatoxins. The present study provides a new method on molecular characterization of A. section Flavi in Turkey.


Toxins ◽  
2019 ◽  
Vol 11 (9) ◽  
pp. 501 ◽  
Author(s):  
Mahror Norlia ◽  
Selamat Jinap ◽  
Mahmud Ab Rashid Nor-Khaizura ◽  
Son Radu ◽  
Cheow Keat Chin ◽  
...  

Peanuts are widely consumed in many local dishes in southeast Asian countries, especially in Malaysia which is one of the major peanut-importing countries in this region. Therefore, Aspergillus spp. and aflatoxin contamination in peanuts during storage are becoming major concerns due to the tropical weather in this region that favours the growth of aflatoxigenic fungi. The present study thus aimed to molecularly identify and characterise the Aspergillus section Flavi isolated from imported peanuts in Malaysia. The internal transcribed spacer (ITS) and β-tubulin sequences were used to confirm the species and determine the phylogenetic relationship among the isolates, while aflatoxin biosynthesis genes (aflR, aflP (omtA), aflD (nor-1), aflM (ver-1), and pksA) were targeted in a multiplex PCR to determine the toxigenic potential. A total of 76 and one isolates were confirmed as A. flavus and A. tamarii, respectively. The Maximum Likelihood (ML) phylogenetic tree resolved the species into two different clades in which all A. flavus (both aflatoxigenic and non-aflatoxigenic) were grouped in the same clade and A. tamarii was grouped in a different clade. The aflatoxin biosynthesis genes were detected in all aflatoxigenic A. flavus while the non-aflatoxigenic A. flavus failed to amplify at least one of the genes. The results indicated that both aflatoxigenic and non-aflatoxigenic A. flavus could survive in imported peanuts and, thus, appropriate storage conditions preferably with low temperature should be considered to avoid the re-emergence of aflatoxigenic A. flavus and the subsequent aflatoxin production in peanuts during storage.


2014 ◽  
Vol 7 (3) ◽  
pp. 335-344 ◽  
Author(s):  
C.N. Ezekiel ◽  
M.C. Adetunji ◽  
O.O. Atanda ◽  
J.C. Frisvad ◽  
J. Houbraken ◽  
...  

In order to facilitate easy and rapid identification of aflatoxin-producing Aspergillus species, the phenotypic traits of Aspergillus section Flavi isolates were examined on neutral red desiccated coconut agar (NRDCA). Phenotype variations in colony morphology and the relationship between colour/intensity of fluorescence and aflatoxin production were assessed. The isolates included 10 Aspergillus minisclerotigenes strains, 11 non-aflatoxigenic Aspergillus flavus L strains, 29 aflatoxigenic A. flavus L strains and 20 strains each of Aspergillus parasiticus and Aspergillus parvisclerotigenus. The NRDCA medium supported morphological differentiation of the four species based on colony features, conidia type and colour. In particular, the two very closely related minisclerotial species, A. minisclerotigenes and A. parvisclerotigenus, were clearly differentiated by their colony colour on NRDCA. All toxigenic isolates produced aflatoxins in the culture medium in varying quantities. Plates of aflatoxigenic A. flavus L strains fluoresced bluish purple/lavender around the colony on the obverse and pastel blue on the reverse side due to aflatoxin B production while those of A. minisclerotigenes, A. parasiticus and A. parvisclerotigenus fluoresced with a light blue or light turquoise ring around the colony on the obverse and light sky blue or cadet blue on the reverse side depending on the amount of aflatoxin B and G produced. The colour of fluorescence significantly correlated (r=0.95, P=0.001) with the type(s) of aflatoxins produced by the isolates. In addition, the concentration of aflatoxins significantly (r=0.92; P=0.001) influenced the intensity of fluorescence in the aflatoxin-producing species. NRDCA can therefore be used for the rapid identification of Aspergillus section Flavi species based on colonial characteristics, and grouping of species into B and B+G aflatoxin producers within 5 days thus obviating the need for chemical analysis of the culture.


2009 ◽  
Vol 2 (3) ◽  
pp. 263-277 ◽  
Author(s):  
J. Varga ◽  
J. Frisvad ◽  
R. Samson

Aflatoxins are decaketide-derived secondary metabolites which are produced by a complex biosynthetic pathway. Aflatoxins are among the economically most important mycotoxins. Aflatoxin B1 exhibits hepatocarcinogenic and hepatotoxic properties, and is frequently referred to as the most potent naturally occurring carcinogen. Acute aflatoxicosis epidemics occur in several parts of Asia and Africa leading to the death of several hundred people. Aflatoxin production has incorrectly been claimed for a long list of Aspergillus species and also for species assigned to other fungal genera. Recent data indicate that aflatoxins are produced by 13 species assigned to three sections of the genus Aspergillus: section Flavi (A. flavus, A. pseudotamarii, A. parasiticus, A. nomius, A. bombycis, A. parvisclerotigenus, A. minisclerotigenes, A. arachidicola), section Nidulantes (Emericella astellata, E. venezuelensis, E. olivicola) and section Ochraceorosei (A. ochraceoroseus, A. rambellii). Several species claimed to produce aflatoxins have been synonymised with other aflatoxin producers, including A. toxicarius (=A. parasiticus), A. flavus var. columnaris (=A. flavus) or A. zhaoqingensis (=A. nomius). Compounds with related structures include sterigmatocystin, an intermediate of aflatoxin biosynthesis produced by several Aspergilli and species assigned to other genera, and dothistromin produced by a range of non-Aspergillus species. In this review, we wish to give an overview of aflatoxin production including the list of species incorrectly identified as aflatoxin producers, and provide short descriptions of the 'true' aflatoxin producing species.


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