Survey of organochlorine-tolerant culturable mycota from contaminated soils, and 2,4-D removal ability of Penicillium species in synthetic wastewater.

Agrochemical wastewater, which is produced by the extensive use of herbicides, has become a serious environmental pollutant. In this study, culturable mycota were isolated from soils contaminated with herbicides 2,4-dichlorophenoxyacetic acid (2,4-D) and 4-chloro-2-methylphenoxyacetic acid (MCPA), and their ability to tolerate and remove 2,4-D was assessed. The mycota were isolated on solid medium supplemented with 10 mmol L-1 of MCPA or 2,4-D. Tolerance and removal assays were performed in synthetic wastewater, and removal was quantified by HPLC-UV and MS/MS. Fusarium spp., Aspergillus spp., and Penicillium spp. were the most frequently isolated genera. Six Penicillium strains were able to tolerate up to 25 mmol L-1 of 2,4-D. Within this group, two P. crustosum strains (RCP4 and RCP13) degraded more than 50% of the 2,4-D in the medium during the first 7 days of incubation. Removal percentages reached 54% for RCP4 and 75% for RCP13 after 14 days. These two strains, therefore, could potentially be considered for the design of bioaugmentation strategies aimed at reducing contamination by 2,4-D in wastewater.

Influence of a glyphosate-based herbicide on growth parameters and aflatoxin B1 production by Aspergillus section Flavi on maize grains.

Glyphosate-based herbicides (GBH) are the main pesticides applied worldwide on maize production. Glyphosate-resistant weeds led to the repeated application of high doses of the pesticide. In addition to environmental conditions, the presence of GBH affects the development of Aspergillus species and aflatoxin B1 (AFB1) production under in vitro conditions. The aim of this work was to evaluate the influence of a commercial GBH on growth and AFB1 production by Aspergillus flavus and Aspergillus parasiticus strains under different water activity (aW) conditions. The following concentrations of active ingredient glyphosate were evaluated: 20, 50, 200 and 500mM. The lag phase prior to growth and growth rate did not change at 20 and 50mM (that is, at field recommended doses) at 0.98 and 0.95 aW; however, at increasing GBH concentrations, between 200 and 500mM, the growth rate decreased at all aW conditions. In general, as the GBH concentration increased, AFB1 production decreased. However, a significant increase in toxin accumulation was found only at one of the aW conditions (0.95) at 21 days with 50mM of GBH in A. flavus and 20 and 50mM of GBH in A. parasiticus. These results show that, even though Aspergillus section Flavi growth did not increase, AFB1 production increased on maize grains at GBH concentrations similar to those of field recommended doses under favorable water availability and temperature conditions.

Isolation of culturable mycota from Argentinean soils exposed or not-exposed to pesticides and determination of glyphosate tolerance of fungal species in media supplied with the herbicide / Aislamiento de la microbiota cultivable de suelos argentinos expuestos y no expuestos a plaguicidas y determinación de la tolerancia a glifosato de especies fúngicas en medios suplementados con el herbicida

Abstract The current agricultural system has led to the development of glyphosate (GP)-resistant weeds, causing an increase in GP doses and applications. Native mycota of pesticide-contaminated sites are the major source of pesticide-degrading microorganisms. The aims of the present study were to isolate the GP-tolerant culturable mycota in two soils with different pesticide exposure from Córdoba, Argentina, and to evaluate the growth parameters in native fungal isolates in the presence of GP and the effective dose that caused 50% growth reduction. The results showed that the genera Fusarium, Aspergillus, Mucor, Penicil-lium and Sterilia were the prevalent fungi isolated from soils both exposed and not-exposed to pesticides. The highest value (>100 mM) of effective concentration of herbicide that caused 50% growth inhibition (EC50), was found for Trichoderma isolates. Sterilia spp. had EC50 val-ues of 100mM, while Aspergillus spp. and Mucor had EC50 values between 50 and 100mM. The growth rate evaluation varied according to the isolates and GP concentrations. The data showed that all Aspergillus spp., Trichoderma spp., Mucor and three Sterilia spp. had the best growth performance in media supplied with GP after a variable acclimation period. This study provides valuable data for further studies that would allow to know the metabolic capacity of these fungal species that can be potential candidates for GP removal from contaminated environments.

Resumen El sistema agrícola actual condujo ai desarrollo de malezas resistentes a glifos-ato (GP), lo que deriva en el aumento de las dosis y el número de aplicaciones requeridas. La microbiota nativa de sitios contaminados con plaguicidas constituye la fuente principal de microorganismos capaces de degradar este herbicida. Los objetivos del presente estudio fueron aislar la microbiota cultivable tolerante a GP a partir de dos suelos agrícolas con diferente grado de exposición a plaguicidas, ambos de la provincia de Córdoba, Argentina, y evaluar en los hongos nativos aislados los parámetros de crecimiento en presencia de GP y la dosis efectiva que produce un 50% de reducción del crecimiento (EC50). Los géneros Fusarium, Aspergillus, Mucor, Penicillium y Sterilia fueron los de mayor prevalencia, tanto en suelos expuestos como no expuestos a plaguicidas. La mayor EC50 hallada (> 100 mM) correspondió a dos aislamientos de Trichoderma. En dos aislamientos de Sterilia, los valores de EC50 fueron de 100 mM, mientras que en los dos de Aspergillus y en uno de Mucor fueron de entre 50 y 100 mM. La velocidad de crecimiento varió según la concentración de GP empleada y el aislamiento considerado. Todos los aislamientos de Aspergillus, Trichoderma y Mucor, y tres aislamientos de Sterilia mostraron el mejor rendimiento en cuanto al crecimiento en los medios con GP después de un período de aclimatación variable. Este trabajo proporciona datos preliminares valiosos referidos a la capacidad metabólica de estos aislados y su potencial para remover GP en ambientes contaminados.

Herbicides based on 2,4-D: its behavior in agricultural environments and microbial biodegradation aspects. A review.

One of the main herbicides used in the agricultural environments is 2,4-dichlorophenoxyacetic acid (2,4-D). It is a synthetic plant hormone auxin employed in many crops including rice, wheat, sorghum, sugar cane, and corn to control wide leaf weeds. The indiscriminate use of pesticides can produce numerous damages to the environment. Therefore, this review has the objective to provide an overview on the main characteristics of the herbicides based on 2,4-D, mostly on the role of microorganisms in its degradation and its main degradation metabolite, 2,4- dichlorophenol (2,4-DCP). The remediation processes carried out by microorganisms are advantageous to avoid the pollution of the environment as well as to safeguard the population health.

Isolation of culturable mycota from Argentinean soils exposed or not-exposed to pesticides and determination of glyphosate tolerance of fungal species in media supplied with the herbicide.

The current agricultural system has led to the development of glyphosate (GP)-resistant weeds, causing an increase in GP doses and applications. Native mycota of pesticide-contaminated sites are the major source of pesticide-degrading microorganisms. The aims of the present study were to isolate the GP-tolerant culturable mycota in two soils with different pesticide exposure from Córdoba, Argentina, and to evaluate the growth parameters in native fungal isolates in the presence of GP and the effective dose that caused 50% growth reduction. The results showed that the genera Fusarium, Aspergillus, Mucor, Penicillium and Sterilia were the prevalent fungi isolated from soils both exposed and not-exposed to pesticides. The highest value (>100mM) of effective concentration of herbicide that caused 50% growth inhibition (EC50), was found for Trichoderma isolates. Sterilia spp. had EC50 values of 100mM, while Aspergillus spp. and Mucor had EC50 values between 50 and 100mM. The growth rate evaluation varied according to the isolates and GP concentrations. The data showed that all Aspergillus spp., Trichoderma spp., Mucor and three Sterilia spp. had the best growth performance in media supplied with GP after a variable acclimation period. This study provides valuable data for further studies that would allow to know the metabolic capacity of these fungal species that can be potential candidates for GP removal from contaminated environments.

Effect of in vitro glyphosate on Fusarium spp. growth and disease severity in maize.

BACKGROUND: Glyphosate (GP) is one of the main pesticides used for maize production. Fusarium sp. is a fungal genus with several phytopathogenic species and toxigenic features. In this study, the culturable soil mycota was evaluated using the surface-spray method. The effect of GP on the growth parameters (growth rate and lag phase) of Fusarium spp. was also tested on solid media conditioned with different water activities. Finally, the GP effect on disease severity caused by Fusarium sp. in maize seedlings was studied. RESULTS: The results showed that Fusarium species are frequently isolated from GP-exposed soils. The GP concentrations tested had a significant effect on F. graminearum, F. verticillioides and F. oxysporum growth parameters on solid media. The pathogenicity tests showed that the disease severity of the maize seedlings significantly increased with increasing GP concentrations. CONCLUSIONS: This study showed that Fusarium species are frequently isolated from pesticide-exposed soils and the GP concentrations tested had a significant effect both on growth parameters and disease severity in maize. This study provides an approach to the effect of GP on Fusarium sp. growth and pathogenicity that reinforces the importance of evaluating all the factors that could affect feed and food production. © 2019 Society of Chemical Industry.

Effect of atrazine on growth and production of AFB1 in Aspergillus section Flavi strains isolated from maize soils.

Atrazine is one of the most frequently used herbicides in Argentina for controlling broadleaf weeds and annual grasses. Currently, there is limited information on the impact of triazine herbicides on mycotoxin production and growth parameters of toxigenic fungi in maize. The objective of this study was to evaluate the effect of different concentrations of atrazine on the lag phase prior to growth, the growth rate, and on production of aflatoxin B1 (AFB1) of Aspergillus flavus and Aspergillus parasiticus strains, on maize meal extract agar (MMEA) under different water activities (aW) and temperatures. A commercial formulation of atrazine was added to MMEA medium at 0, 5, 10, 50, or 100 mmol/l, adjusted to 0.98, 0.95, and 0.93 aW, and incubated at 28 °C and 37 °C for 21 days. AFB1 was determined by HPLC after 7, 14, and 21 days of incubation. In the control treatments, a significant increase in the time prior to growth was observed and as the aW decreased, at both temperatures, the growth rate of the strains also decreased. A significant increase in growth rate was observed as the concentration of atrazine in the medium increased, for all aW levels tested. The optimal conditions for the accumulation of AFB1 in the control treatments were 0.98 aW and 28 °C, after 7 days of incubation. As the concentration of herbicide increased, AFB1 production also increased (P < 0.05). These results add to the knowledge about consequences with regard to aflatoxin production of the use of excessive atrazine doses in extensive maize culture.

Survey of Aspergillus and Fusarium species and their mycotoxins in raw materials and poultry feeds from Córdoba, Argentina.

The aims of the present work were: (1) to determine both mycobiota in raw materials and finisher poultry feed, as well as the ability to produce aflatoxin B1 by A. flavus strains, and (2) to evaluate the natural co-occurrence of aflatoxins (AFs), fumonisins (FBs), gliotoxin, diacetoxyscirpenol (DAS), HT-2 toxin, and T-2 toxin in poultry feed by LC-MS/MS. Nineteen percent of raw materials and 79% of finisher poultry feed samples exceeded the maximum allowed total fungal count (1 × 10(4) CFU g(-1)) to ensure hygienic quality. Aspergillus flavus was the only species belonging to section Flavi which was isolated while Fusarium verticilliodes was the prevalent species. Forty-seven percent of A. flavus strains were aflatoxin B1 producers and the highest frequency of aflatoxigenic strains was isolated from finisher poultry feeds. Principal component analysis showed that corn grains are closely related with total fungal and Fusarium counts. This positive relationship suggests that total fungal and Fusarium spp. counts in poultry feed might come mainly from corn grains. Regarding poultry feeds, in ground finisher type, Aspergillus spp. counts increased as water activity (aw) diminished. A positive relationship among aw, total fungal and Fusarium spp. counts was observed in both ground finisher and ground starter feed. Several mycotoxins were monitored in feeds by applying the LC MS/MS technique. One hundred percent of poultry samples were contaminated with FB1, and the highest levels were detected in pelleted finisher poultry. AFB1, gliotoxin, DAS, HT-2 toxin, and T-2 toxin were not detected in any poultry feed. The scarcity of available mycotoxicological studies from Argentinean poultry feed using a multitoxin analysis technique enhances the contribution of the findings of this report.