Niger Aggregate Strains Research Articles

Assessment of growth of Aspergillus spp. from agricultural soils in the presence of glyphosate

Agriculture is one of the bases of the Argentine economy. Glyphosate is undoubtedly one of the most important herbicides used. The increasing consumption and the efficiency of glyphosate-based herbicides have encouraged several studies on their persistence in soils, their effects on soil microbiota and their degradation processes. Fungi have been reported as being the main herbicide-degrading microorganisms as well as the most tolerant to environmental stress conditions. This study evaluated the growth performance of Aspergillus section Flavi and Aspergillus niger aggregate strains on Czapek Dox media supplied with a commercial glyphosate formulation as sole source of carbon (CZC), phosphorus (CZP) or nitrogen (CZN). Six Aspergillus spp. strains were evaluated. Each medium was stab-inoculated with fungal spores from 7-day old cultures. Two measures of colony radii were taken daily. All of the Aspergillus section Flavi strains showed a significant increase (from 24 to 44%) in growth rate on the CZN medium, as compared to controls. The A. niger aggregate strains exhibited the same behavioral pattern under all the conditions tested, except on the CZN medium. Velutinous or slightly floccose colonies with abundant sporulation were observed on CZP. Moreover, the colonies produced sparse sporulation on CZC or CZN media, being their appearances completely different from those on the CZP medium. This study establishes that A. section Flavi and A. niger aggregate strains can grow in vitro in the presence of glyphosate, especially when it is used as a sole source of phosphorus or nitrogen.

Influence of the pesticides glyphosate, chlorpyrifos and atrazine on growth parameters of nonochratoxigenic Aspergillus section Nigri strains isolated from agricultural soils

This investigation was undertake to determine the effect of glyphosate, chlorpyrifos and atrazine on the lag phase and growth rate of nonochratoxigenic A. niger aggregate strains growing on soil extract medium at −0.70, −2.78 and −7.06 MPa. Under certain conditions, the glyphosate concentrations used significantly increased micelial growth as compared to control. An increase of about 30% was observed for strain AN 251 using 5 and 20 mg L−1 of glyphosate at −2.78 MPa. The strains behaved differently in the presence of the insecticide chlorpyrifos. A significant decrease in growth rate, compared to control, was observed for all strains except AN 251 at −2.78 MPa with 5 mg L−1. This strain showed a significant increase in growth rate. With regard to atrazine, significant differences were observed only under some conditions compared to control. An increase in growth rate was observed for strain AN 251 at −2.78 MPa with 5 and 10 mg L−1 of atrazine. By comparison, a reduction of 25% in growth rate was observed at −7.06 MPa and higher atrazine concentrations. This study shows that glyphosate, chlorpyrifos and atrazine affect the growth parameters of nonochratoxigenic A. niger aggregate strains under in vitro conditions.

Survey of Aspergillus section Flavi presence in agricultural soils and effect of glyphosate on nontoxigenic A. flavus growth on soil-based medium

To evaluate the cultivable mycobiota from agricultural soils exposed to pesticides, the aflatoxigenic capacity of Aspergillus section Flavi strains and the effect of glyphosate on lag phase and growth rates of native nontoxigenic Aspergillus flavus under different water potential (MPa) conditions on soil-based medium. Culturable mycobiota analysis from different agricultural soils was performed by the surface spread method. The effect of glyphosate (0-20 mmol l(-1)) on the growth of A. flavus strains was evaluated on a soil extract solid medium. Mycobiota analysis of crop soils showed the presence of twenty-one genera of filamentous fungi. Aspergillus flavus and Aspergillus niger aggregate strains were isolated from the three soil types. Ninety-two per cent of A. flavus strains were toxigenic. In vitro assay results showed that at -0·70 MPa, a significant increase in growth rate in all strains was recorded at 5 and 20 mmol l(-1) of glyphosate. At -2·78 MPa, this parameter remained constant at all glyphosate concentrations, except in GM4 strain where an increase in growth rate was recorded with increasing pesticide concentrations. At -7·06 MPa, a significant increase in growth rate has also been observed in GM 3 strain with 5 mmol l(-1) and in GM 4 strain with 10 and 20 mmol l(-1). This study showed that the imperfecti fungi Aspergillus spp., Penicillium spp., Trichoderma spp., Cladosporium spp. and Paecilomyces spp. are isolated as prevalent groups in agricultural soil exposed to pesticides, and the capacity of nontoxigenic A. flavus strains to tolerate different glyphosate concentrations under different water potential (MPa) conditions. This manuscript makes a contribution to the knowledge of cultivable fungal populations from agricultural soils exposed to pesticides and the glyphosate tolerance of A. flavus strains.

Effect of Temperature on Production of Ochratoxin A by Aspergillus niger in Orange Juice

This challenging study was carried out to evaluate the temporal production of ochratoxin A (OTA) by Aspergillus niger ATCC 16404 and wild type A. niger AM at different temperatures in fresh squeezed orange juice (Citrus sinensis [L.] Osbeck cv Tarocco). Each strain, inoculated into the filtered orange juice, was incubated at 4°C, 20°C, and 26°C for 28 days. In the juice, at 26°C and 20°C, the concentration of A. niger ATCC 16404 increased by more than 2 log10 up to the 21st day. At 4°C it remained constant. The microbial load of A. niger AM decreased at all temperatures. At 26°C, the maximum OTA accumulation found was 3.44 ng/mL on the 21st day for A. niger ATCC 16404 and 8. 44 ng/mL on the 7th day for A. niger AM. The OTA synthesis seemed to be an intrinsic strain-dependent mechanism. A. niger ATCC 16404 produced OTA in accordance with the higher temperatures and the biomass concentrations, whereas A. niger AM produced the toxin at all temperatures regardless of its biomass. These results showed that fresh orange juice contaminated with Aspergillus niger aggregate strains may contain OTA levels.

Effect of indigenous mycobiota on ochratoxin A production by Aspergillus carbonarius isolated from soil

This study determined the biotic interaction between 30 non-toxigenic indigenous strains of Aspergillus niger aggregate, Aspergillus flavus, Trichoderma spp., Mucor spp., Cladosporium spp., Ulocladium spp., Curvularia spp., Absidia spp., Geotrichum spp. and Acremonium spp., isolated from soil destined for maize crops, with respect to their ability to prevent ochratoxin A (OTA) production by A. carbonarius on "in vitro" assay, on liquid and solid medium. OTA production was completely inhibited when A. carbonarius was inoculated in a interactive mixed culture with all A. niger aggregate strains assayed, a 80% of Trichoderma spp. strains, a 40% of Cladosporium spp. strains, Acremonium spp and Geotrichum spp; only one strain of A. flavus tested was able to completely inhibit the mycotoxin accumulation. OTA production increased when A. carbonarius ACS 8 was growing on liquid interactive mixed culture with Mucor spp strains. These results demonstrated that OTA production by Aspergillua carbonarius strain was significantly influenced by the presence of different non-toxigenic fungal strains when growing together on paired cultures.

Mycotoxin production by Aspergillus niger aggregate strains isolated from harvested maize in three Portuguese regions

BackgroundMaize is considered one of the crops more susceptible to mycotoxins in the world. Two of the mycotoxins commonly associated with maize are fumonisins and ochratoxin A. Aspergillus niger is a known producer of ochratoxin A and is easily found in maize. Recently, however, A. niger has been reported to produce as well fumonisins, mainly fumonisin B2. AimsThe aim of this study was to isolate A. niger strains from maize samples collected in three Portuguese maize growing regions and to detect the production of both fumonisin B2 and ochratoxin A. MethodsNinety five maize samples were collected, plated, and all observable Aspergillus section Nigri strains were isolated. Strains were morphologically characterized and mycotoxin production was determined by HPLC-FD. ResultsIsolations resulted in a total of 270 strains of black Aspergillus from 73 samples (77% of the samples). About 14% of those strains were found to produce ochratoxin A and 39% of the strains were found to produce fumonisin B2. ConclusionsAn association between the production of these two mycotoxins could not be found and no conclusions could be taken whether the presence of A. niger aggregate strains will increase the risk of maize contamination with fumonisins and more specifically with fumonisin B2.

Evaluation of aflatoxin B1 and ochratoxin A in interacting mixed cultures of Aspergillus sections Flavi and Nigri on peanut grains

The influence of inoculum size on the colony-forming units, production of aflatoxin B1 (AFB1) and ochratoxin A (OTA) was determined when Aspergillus flavus and A. niger aggregate strains were cultured alone and in pairs on irradiated peanut grains at 28°C and 0.97 water activity (aW). The results showed a marked influence of inoculum factor on fungal counts, AFB1 and OTA production in single and paired cultures. Fungal counts of the A. niger aggregate strain in interacting cultures at 7, 14 and 21days of incubation were significantly higher than those observed in the A. flavus strain, except in the mixed culture with 10(2) spores/ml of both strains. In all mixed culture assays, the AFB1 production was significantly reduced in comparison with the accumulation of mycotoxin in single cultures. A total inhibition in AFB1 production was observed in some interactions as 10(2) spores/ml of A. flavus and 10(3) spores/ml of A. niger aggregate strain at 7 and 14days, among others. With regard to OTA production, a stimulation in the interacting cultures was observed at all inoculum sizes and incubation period. The highest levels of OTA accumulation were observed at 14days for all interacting cultures. The maximum level was reach in the culture 10(3) spores/ml of A. niger aggregate and 10(4) spores/ml of A. flavus (p < 0.001). These results suggest that, under optimal environmental conditions in peanut grains, the interaction between A. flavus and A. niger aggregate strains could result in an inhibition of AFB1 and in a stimulation of OTA production.

Development of a Real Time PCR system for detection of ochratoxin A-producing strains of the Aspergillus niger aggregate

Members of Aspergillus section Nigri are distributed worldwide, being considered as common food spoilage fungi. Some species of this section produce ochratoxin A (OTA), mainly Aspergillus carbonarius and several members of the Aspergillus niger aggregate. Detection of ochratoxigenic A. niger aggregate strains is important to prevent OTA contamination in foodstuffs. A new Real Time PCR procedure has been developed for the rapid and specific detection and quantification of ochratoxin A-producing strains of the A. niger aggregate. Two specific primers delimiting a 120 bp fragment and a probe were designed and directed to a polyketide synthase (PKS) from A. niger CBS 513.88 genome. This PKS has a strong similarity to PKS of A. ochraceus fragment involved in ochratoxin biosynthesis. Specificity was confirmed by testing primers towards purified DNA from 91 fungal strains, including reference and food isolates. The SYBR-Green and the TaqMan approaches developed allowed the specific detection only of ochratoxigenic strains of the A. niger aggregate. All other analyzed food related fungi gave negative results. This is the first report on a Real Time PCR system for the detection of OTA-producing strains of the A. niger aggregate.

Early detection of Aspergillus carbonarius and A. niger on table grapes: a tool for quality improvement

Aspergillus carbonarius and A. niger aggregate are the main fungal contaminants of table grapes. Besides their ability to cause black rot, they can produce ochratoxin A (OTA), a mycotoxin that has attracted increasing attention worldwide. The objective of this work was to set up a simple and rapid molecular method for the early detection of both fungi in table grapes before fungal development becomes evident. Polymerase chain reaction (PCR)-based assays were developed by designing species-specific primers based on the polyketide synthases (PKSS) sequences of A. carbonarius and A. niger that have recently been demonstrated to be involved in OTA biosynthesis. Three table grape varieties (Red globe, Crimson seedless, and Italia) were inoculated with A. carbonarius and A. niger aggregate strains producing OTA. The extracted DNA from control (non-inoculated) and inoculated grapes was amplified by PCR using ACPKS2F-ACPKS2R for A. carbonarius and ANPKS5-ANPKS6 for A. niger aggregate. Both primers allowed a clear detection, even in symptomless samples. PCR-based methods are considered to be a good alternative to traditional diagnostic means for the early detection of fungi in complex matrix for their high specificity and sensitivity. The results obtained could be useful for the definition of a ‘quality label’ for tested grapes to improve the safety measures taken to guarantee the production of fresh table grapes.

Food-grade antioxidants and antimicrobials to control growth and ochratoxin a production by Aspergillus section Nigri on peanut kernels.

Each year, a significant portion of the peanuts produced cannot be marketed because of fungal disease at the postharvest stage and mycotoxin contamination. Antioxidants could be used as an alternative to fungicides to control ochratoxigenic fungi in peanuts during storage. This study was carried out to determine the effect of the antioxidant butylated hydroxyanisole (BHA) and the antimicrobial propyl paraben (PP) on the lag phase before growth, growth rate, and ochratoxin A (OTA) production by Aspergillus section Nigri strains in peanut kernels under different conditions of water activity (aw) and temperature. At 20 mM/g BHA, 18 degrees C, and 0.93 aw, complete inhibition of growth occurred. For PP, there was no growth at 20 mM/g, 18 degrees C, and 0.93, 0.95, and 0.98 aw. BHA at 20 mM/g inhibited OTA production in peanuts by Aspergillus carbonarius and Aspergillus niger aggregate strains at 0.93 aw and 18 degrees C. PP at 20 mM/g completely inhibited OTA production at 18 degrees C. The results of this work suggest that PP is more appropriate than BHA for controlling growth and OTA production by Aspergillus section Nigri species in peanut kernels.

Aspergillus section Nigri species isolated from different wine-grape growing regions in Argentina

Vineyards located in eight grape growing regions of Argentina during the harvest season 2006/07 were evaluated. The aims of this study were to determine the incidence of Aspergillus section Nigri, their ability to produce ochratoxin A (OTA) and to evaluate the OTA natural occurrence in grapes. Bunches of grapes at maturation stage were collected, and grapes (50 per sample) were plated on Petri dishes containing dichloran-glycerol 18% agar (DG18) and dichloran-rose bengal-chloramphenicol agar (DRBC) media. After an incubation period of 7 days at 25 °C, the mycoflora belonging to Aspergillus section Nigri was identified. OTA occurrence and the toxicogenic ability of the strains were analyzed by HPLC. A. niger aggregate strains were dominant showing the highest infection percentage (81%), followed by A. carbonarius (11%) and Aspergillus uniseriate (8%). A. carbonarius strains presented the highest percentage of OTA-producer strains (82%) and the highest toxin levels (mean 202 ng/g). A positive correlation between the isolation percentage of A. carbonarius in grapes and temperature was found. The warmest regions showed the highest A. carbonarius incidence. OTA was detected at low levels in grapes during the survey. OTA levels in grapes and rain at harvest time correlated positively.

Impact of some environmental factors on growth and production of ochratoxin A of/by Aspergillus tubingensis, A. niger, and A. carbonarius isolated from moroccan grapes

The effects of temperature, water activity (aw), incubation time, and their combinations on radial growth and ochratoxin A (OTA) production of/by eight Aspergillus niger aggregate strains (six A. tubingensis and two A. niger) and four A. carbonarius isolated from Moroccan grapes were studied. Optimal conditions for the growth of most studied strains were shown to be at 25 degrees C and 0.95 aw. No growth was observed at 10 degrees C regardless of the water activity and isolates. The optimal temperature for OTA production was in the range of 25 degrees C-30 degrees C for A. carbonarius and 30 degrees C-37 degrees C for A. niger aggregate. The optimal aw for toxin production was 0.95-0.99 for A. carbonarius and 0.90-0.95 for A. niger aggregate. Mean OTA concentration produced by all the isolates of A. niger aggregate tested at all sampling times shows that maximum amount of OTA (0.24 microg/g) was produced at 37 degrees C and 0.90 aw. However, for A. carbonarius, mean maximum amounts of OTA (0.22 microg/g) were observed at 25 degrees C and 0.99 aw. Analysis of variance showed that the effects of all single factors (aw, isolate, temperature and incubation time) and their interactions on growth and OTA production were highly significant.

Study of the phenotypic and genotypic biodiversity of potentially ochratoxigenic black aspergilli isolated from grapes

Ochratoxin A (OTA) is a mycotoxin with nephrotoxic, carcinogenic, teratogenic and immunotoxic effects, naturally found in agricultural products including grapes and wine. Black Aspergillus species (Section Nigri) are mainly responsible for OTA accumulation in wine grapes and in particular Aspergillus carbonarius and Aspergillus niger aggregate . The biodiversity of potentially ochratoxigenic strains of black aspergilli from different French vineyards in the southern Mediterranean region of Languedoc–Roussillon was studied. One hundred and eighty nine black strains were isolated from grapes and studied according to harvest year, production zone, grape variety and pre-harvest treatment of grapevines. The strains were identified and classified in two groups according to macroscopic and microscopic characters; these were called the A. carbonarius representative group and the A. niger aggregate representative group. Members of each group were classified in subgroups based on macroscopic morphological colony characters. Strain biodiversity was studied according to phenotypic and genotypic characterization and to the OTA production of selected strains on PDA medium. After identification was confirmed by specific PCR using primer pair ITS1/CAR and ITS1/NIG, 24 potential ochratoxigenic strains belonging to A. carbonarius and A. niger aggregate were discriminated by RAPD-PCR using 8 different OPC primers. The use of specific primers supported the identification based on phenotypic and morphological characters. RAPD-PCR patterns demonstrated a considerable diversity among the strains. Clustering among A. niger aggregate strains was associated with production zone and harvest year, but not grape variety or pre-harvest treatment. Clustering among A. carbonarius strains was not associated with any of the above parameters. OTA production of strains on culture medium seemed to correlate better with morphological characters than with genotypic profiles. No clear relation could be established between phenotypic and genotypic characters of the studied black aspergilli.

Influence of ecophysiological factors on growth, lag phase and ochratoxin A production by Aspergillus niger aggregate strains in irradiated corn grains

Ochratoxin A (OTA) is one of the most important mycotoxins of worldwide concern for human health. The aims of this study were to evaluate the effects of water activity, temperature, incubation time and their interactions on growth, lag phase and OTA production by two strains belonging to A. niger aggregate in irradiated corn grains. This study shows that OTA production on corn grains occurs within a more restricted range of water activity and temperature than fungal growth. This point is important in order to maintain the appropriate conditions in stored corn that prevent toxigenic fungal growth and thus OTA production. Irradiated corn grains were re-hydrated to 0.910-0.995 of a(W) with sterile distillate water. The temperatures assayed were 15, 25 and 30 degrees C. Growth assessment was made every day during the incubation period (21 days) to calculate the growth rate, lag phase and the OTA production at 7, 14 and 21 days. Both strains showed optimum growth at 30 degrees C at 0.951 a(W). Ochratoxin A production occurred over a range of temperatures (15-30 degrees C) with optimum production at 25 degrees C depending on a(W) assayed. Maximum OTA production was obtained for both strains at 0.973 a(W). The results obtained suggest that the storage of corn grains at water activities lower than 0.951 and 15 degrees C should prevent these fungal species growth and OTA production during 21 days approximately.

Ecophysiological factor effect on growth rate, lag phase and ochratoxin A production by Aspergillus niger aggregate strains on irradiated peanut seeds

The aims of this study were to evaluate the effects of water activity, temperature, incubation time and their interactions on lag phase, growth rate and ochratoxin A (OTA) production by strains belonging to the Aspergillus niger aggregate on irradiated peanut seeds. In the temperature and water activity range assayed, the optimal conditions of growth for RCP42 and RCP176 strains on irradiated peanut seeds were 0.995 a w and 30 °C being the growth rates of 12.4 and 14.6 mm/day, respectively. The maximum OTA production occurred at 0.973 a w and 25 °C for both strains assayed; whereas the minimum OTA production was obtained at 0.951 a w and 15 °C, at 14 and 21 days of incubation for RCP42 and RCP176 strains, respectively. The amount of OTA accumulated during 21 days assayed by both strains varied from 6.5 to 460 μg/g and from 10 to 210 μg/g with mean levels of 119.2 and 97.5 μg/g for RCP42 and RCP176 strains, respectively. The variance analysis (ANOVA) revealed that OTA concentration produced by RCP42 strain was significantly ( p < 0.0001) greater than that produced by RCP176 strain. If the strains with which the experiments were carried out were representative of the Aspergillus niger aggregate toxigenic species and the water activity in peanut seeds stored at 0.910 or lower was maintained, OTA production would be reduced during at least 21 days at variable temperatures.

OTA-producing fungi isolated from stored cocoa beans

The aim of this study was to identify fungal populations in unroasted cocoa beans stored in Spain in order to evaluate the ochratoxin A (OTA)-production ability of certain Aspergillus isolates. Twenty batches of cocoa beans from different origins and with different OTA content were selected for this study. Three Aspergillus carbonarius and 13 Aspergillus niger aggregate strains isolated from these cocoa bean samples were selected to evaluate their OTA synthesis ability, being the only A. carbonarius isolates which are OTA producers [<limit of detection (LOD) = 3520 microg kg(-1) culture medium; LOD = 6 microg kg(-1) culture medium]. No correspondence was found between the OTA levels in cocoa beans and the presence of OTA-producing fungi. Nonetheless, some samples contained A. carbonarius with a high OTA-producing ability and, consequently, specific fungal controls should be set up during storage to avoid this toxin. Toxigenic fungi in cocoa beans are not well understood. This study attempted to identify these fungi and evaluate their OTA-producing ability.

Utility of Microsatellite Markers and Amplified Fragment Length Polymorphism in the Study of Potentially Ochratoxigenic Black Aspergilli

Microsatellite markers and the results of amplified fragment length polymorphism (AFLP) were compared in the characterization of 68 Aspergillus carbonarius and A. niger aggregate strains of differing ochratoxin-producing ability and from different geographic areas, isolated mainly from grapes and soil. AFLP was applied to both A. carbonarius and A. niger aggregate strains, and it clearly differentiated these species. Microsatellite markers were only applied to A. niger aggregate strains because of the species-specific nature of these markers. Both AFLP and microsatellite marker analyses were able to divide A. niger aggregate strains into the two recognized internal transcribed spacer (ITS)-5.8S rDNA RFLP types, N and T. Clustering of A. niger aggregate strains was similar in both AFLP and microsatellite analyses, yielding an additional separation of N type strains into two groups. Both microsatellite marker and AFLP analyses showed high levels of polymorphism in the A. niger aggregate (index of discriminatory power 0.991 and 1.0, respectively). Of the two techniques, microsatellite marker analysis was quicker and more straightforward to perform. In addition, microsatellite marker analysis is more reproducible, and the results can be expressed as quantitative data, making microsatellite markers a good candidate for use in large-scale studies of genetic diversity in A. niger aggregate species.

Effect of gentian violet on the growth of the N and T RFLP types of the Aspergillus niger aggregate

Taxa included in the Aspergillus niger aggregate are difficult to distinguish by phenotypic characterization. In this work, the effect of gentian violet on the growth of the N and T RFLP types of A. niger aggregate strains has been investigated. In total, 105 strains from different sources and origins, including reference cultures and field isolates were studied. Type N and T RFLP patterns, ochratoxin A production and the effect of different concentrations of gentian violet on the growth were determined in these strains. Forty nine strains belonged to the N type and 56 strains to the T type. Sixteen out of the 105 strains assayed were OTA producers. All the OTA-producing species belonged to the RFLP type N and none of the T type strains was able to produce OTA. Approximately 90% of the N type strains grew in the presence of 25 ppm of gentian violet. Only five N type strains did not grow on this medium. One of these strains was A. niger ATCC 22343, a well documented induced mutant strain and the remaining four strains belonged to the new species A. brasiliensis. On the contrary, all the T type strains failed to grow on this medium after 3 days of incubation (sensitivity 89.79%; specificity 100%). The use of growth in gentian violet as an additional character for classification and identification purposes in this taxonomic group may be useful because no phenotypic methods have yet been found that can distinguish between these species.

Digestion of DNA regions to discriminate ochratoxigenic and non-ochratoxigenic strains in the Aspergillus niger aggregate

Aspergillus strains belonging to the Aspergillus niger aggregate, either isolated from Italian grapes or received from public collections, were analysed in order to discriminate between the ochratoxin A (OTA) producing and the non-producing strains by means of the analysis of Internal Transcribed Spacers (ITS), Intergenic Spacers (IGS) and of a β-tubulin gene portion. A. niger and Aspergillus awamori were identified observing the macro- and microscopic features of the colonies and the strains ochratoxigenicity was evaluated through Thin Layer Chromatography and/or High Performance Liquid Chromatography. PCR amplification of ITS, IGS and β-tubulin gene portion produced 600, 440 and 550 bp amplicons, respectively, in all the analysed strains. The digestion of the IGS amplicon with Hinf I and of the other two amplicons with Rsa I, revealed one main profile type (“T–C–C”) associated with the A. niger aggregate strains which lack the ability to produce OTA.

Effect of pH on ochratoxin A production by Aspergillus niger aggregate species

The effect of pH (2–10) on growth and ochratoxin A (OTA) production by 12 Aspergillus niger aggregate strains was studied in two culture media: Czapek yeast autolysate agar (CYA) and yeast extract sucrose agar (YES), over 30 days. The strains were selected to include different sources, different reported abilities to produce OTA and different ITS-5.8S rDNA RFLP patterns. YES was a better culture medium than CYA for OTA production. In this medium, OTA was produced from pH 2 or 3 to 10 depending on the strain. The results show the ability of A. niger aggregate strains not only to grow, but also to produce OTA over a wide pH range. The results will lead to a better understanding of the role of A. niger aggregate strains in the OTA contamination of several food commodities.