Influence of herbicide glyphosate on growth and aflatoxin B1 production by Aspergillus section Flavi strains isolated from soil on in vitro assay

The effect of six glyphosate concentrations on growth rate and aflatoxin B₁ (AFB₁) production by Aspergillus section Flavi strains under different water activity (a_W) on maize-based medium was investigated. In general, the lag phase decreased as glyphosate concentration increased and all the strains showed the same behavior at the different conditions tested. The glyphosate increased significantly the growth of all Aspergillus section Flavi strains in different percentages with respect to control depending on pesticide concentration. At 5.0 and 10 mM this fact was more evident; however significant differences between both concentrations were not observed in most strains. Aflatoxin B₁ production did not show noticeable differences among different pesticide concentrations assayed at all a_W in both strains. This study has shown that these Aspergillus flavus and A. parasiticus strains are able to grow effectively and produce aflatoxins in high nutrient status media over a range of glyphosate concentrations under different water activity conditions.     

Biological control of AFB1-producing Aspergillus section Flavi strains isolated from brewer's grains,alternative feed intended for swine production in Argentina

The aim of the present study was to investigate the inhibitory activity of lactic acid bacteria (LAB) isolated from brewer's grains on Aspergillus section Flavi growth and aflatoxin B₁ production. The Aspergillus strains tested were inhibited by all the LAB strains assayed. The isolates Lactobacillus brevis B20, P. pentosaceus B86, Lactococcus lactis subsp. lactis B87, L. brevis B131, and Lactobacillus sp. B144 completely suppressed the fungal growth and reduced aflatoxin B₁ production. In conclusion, LAB isolated from brewer's grains show a high inhibitory activity on fungal growth and aflatoxin biosynthesis by Aspergillus flavus and Aspergillus parasiticus. Further studies must be conducted to evaluate the success of in vitro assays under food environment conditions and to elucidate the antifungal mechanism of these strains.     

Inhibitory properties of Enterococcus spp. isolated from faeces of healthy dogs

The aim of the present study was to evaluate the inhibitory effect by the cross-streak method of nine Enterococcus faecium strains isolated from faeces of healthy dogs and their treated and non-treated cell-free supernatant (CFS) by the well-diffusion test on the growth of potentially pathogenic bacteria isolated from clinical cases and aflatoxigenic Aspergillus section Flavi and the consequent aflatoxin B₁ (AFB₁) production. Results obtained from the cross-strake assay showed that E. faecium MF1, GJ18 and GJ40 presented the major inhibitory activity against all pathogenic strains assayed; E. faecium GJ40 produced the larger inhibitory zones (26–27 mm). Well-diffusion test results showed that the majority of the enterococci strains CFS had antimicrobial activity against the pathogenic microorganisms, especially on Gram negative indicators. Cell-free supernatant of E. faecium GJ40 was the one that produced the largest inhibition zones (14 to 21 mm) in the majority of the indicator microorganisms assayed. All supernatants treated with 10 N NaOH (pH6) showed no inhibitory effect on the indicator strain assayed. With respect to fungal inhibition, any of the CFS assayed significantly inhibited the Aspergillus strains growth. But, in general, all CFS reduced AFB₁ production from 8 to 87%. The results demonstrate that enterococci isolated from healthy dog feaces produce substances with the capacity to inhibit some potential pathogenic bacteria growth and the capacity of inhibiting or reducing the AFB₁ production in vitro.     

Concentrations and Diversity of Microbes from Four Local Bioaerosol Emission Sources in Finland

ABSTRACT

Microbial particles can readily be released into the air from different types of man-made sources such as waste operations. Microbiological emissions from different biological sources and their dispersion may be an issue of concern for area planning and for nearby residents. This study was designed to determine the concentrations and diversity of microbiological emissions from four different man-made source environments: waste center with composting windrows, sewage treatment plant, farming environment, and cattle manure spreading. Samples of airborne particles were collected onto polyvinyl chloride filters at three distances along the prevailing downwind direction, from each source environment during a period of approximately 1 week. These samples were analyzed for 13 species or assay groups of fungi, bacterial genus Streptomyces, and Gram-positive and -negative bacteria using quantitative polymerase chain reaction (PCR). Samples for determining the concentrations of viable fungi and bacteria were collected from all environments using a six-stage impactor. The results show that there were variations in the microbial diversity between the source environments. Specifically, composting was a major source for the fungal genera Aspergillus and Penicillium, particularly for Aspergillus fumigatus, and for the bacterial genus Streptomyces. Although the microbial concentrations in the sewage treatment plant area were significantly higher than those at 50 or 200 m distance from the plant area, in the farming environment or cattle manure spreading area, no significant difference was observed between different distances from the source. In summary, elevated concentrations of microbes that differ from background can only be detected within a few hundred meters from the source. This finding, reported earlier for culturable bacteria and fungi, could thus be confirmed using molecular methods that cover both culturable and nonculturable microbial material.

IMPLICATIONS Concentrations and diversity of airborne microbes increase due to particle emissions from different biological waste treatment applications. However, these emissions cannot be separated from the background concentrations after more than a few hundred meters from the source. As part of a risk assessment, it may be necessary to confirm the behavior of microbial emissions from a specific source. Quantitative PCR is a useful tool for estimating total concentrations of different microbial species or groups as it detects both culturable and nonculturable microbial material.     

Tolerance to agricultural pesticides of strains belonging to four genera of Rhizobiaceae

In order to determine their tolerance to pesticides, 122 strains of rhizobia isolated from different geographical regions, and belonging to the genera Rhizobium, Mesorhizobium, Sinorhizobium and Bradyrhizobium were tested against eight herbicides, four fungicides and five insecticides. Sensitivity to the pesticides was measured by using the filter paper disk method at four concentrations, 0.45, 4.5, 45 and 450 μg per disk. When the pesticides were used at 0.45 μg per disk, a concentration similar to that found when pesticides are applied under field conditions, no inhibition was observed. Strains growth was affected at concentrations of 45 and 450 μg pesticide per disk. These higher concentrations can be encountered when seeds are treated with pesticides. Pesticides tolerance level was correlated to pesticide function, i.e rhizobial strains were more tolerant to insecticides, followed by herbicides and then fungicides. Two fungicides, captan and mancozeb, inhibited the highest number of strains. Only one insecticide, carbaryl, affected the growth of some rhizobial strains. Strains isolated from the arctic (Mesorhizobium spp. and R. leguminosarum bv. viciae), a putative pesticides-free environment, were either less or equally affected by pesticides compared to strains isolated from agricultural regions.     

Effects of glyphosate on the non-target leaf beetle Cerotoma arcuata (Coleoptera: Chrysomelidae) in field and laboratory conditions

This study aimed to assess the glyphosate application effects on the Cerotoma arcuata Oliver (Coleoptera: Chrysomelidae) population in glyphosate-resistant soybean crops. Field studies were conducted with glyphosate and the insecticide endosulfan to observe the effects of these pesticides on C. arcuata, on its damages in the crop and on the populations of natural enemies in glyphosate-resistant soybean crops. Moreover, the lethal and behavioral sublethal response of C. arcuata to glyphosate and endosulfan was conducted in the laboratory. The results of the field and laboratory experiments showed that glyphosate caused moderate toxicity and high irritability in C. arcuata and that endosulfan caused high toxicity and irritability. Therefore, the direct effect of glyphosate on C. arcuata was negative and does not explain the population increases of this pest in glyphosate-resistant soybean. However, the glyphosate also decreased the density of predators. Thus, the negative effect of glyphosate on the predators may be related to population increases of C. arcuata in glyphosate-resistant soybean crops, however, more studies are needed to better evidence this relationship. This study suggests that glyphosate can impact other non-target organisms, such as herbivorous insects and natural enemies and that the use of this herbicide will need to be carefully stewarded to prevent potential disturbances in beneficial insect communities in agricultural systems.     

Pesticides impact on Clavibacter michiganensis ssp. sepedonicus biofilm formation

The effect of various pesticides on the biofilm formation by the phytopathogenic bacterium Clavibacter michiganensis ssp. sepedonicus (Cms), the potato ring rot causative agent, was explored for the first time. Systemic herbicides: 2,4-D, diuron, glyphosate, clopyralid, fluorodifen, as well as the commercial preparations “Lazurite,” “Ridomil Gold,” and the mitochondria inhibiting pesticides analog, sodium monoiodoacetate, were studied. These pesticides' effect on the Cms biofilm formation was shown to be distinct and dependent on the agent under question. Cms biofilm formation was reduced when exposed to sodium monoiodoacetate, as well as “Lazurite” preparation, that could be due to the bactericidal effect of these agents. 2,4-D and “Ridomil Gold” preparation stimulated the biofilm formation. Systemic herbicides diuron, glyphosate, clopyralid, fluorodifen did not exert appreciable influence on the process of bacterial biofilm formation.     

Airborne fungi in low and high allergic prevalence child care centers

Fungi exposure has been linked to asthma and allergies among children. To determine the association between fungal exposure and wheeze and rhinitis symptoms, we examined concentrations of culturable indoor and outdoor fungi of various aerodynamic sizes in low and high allergic prevalence child care centers (CCCs) in Singapore. Environmental parameters were also performed for air temperature, relative humidity and ventilation rates, while information on CCC characteristics was collected via an inspection. Most commonly recovered fungi were Penicillium, Aspergillus, Geotrichum, Cladosporium and sterile mycelia with Geotrichum and sterile mycelia amounting to an average of 71.5% of the total airborne culturable fungi studied. Indoor and outdoor total culturable fungi concentrations and those in the size range of 1.1–3.3 μm were significantly higher in high allergic prevalence CCCs. When fungal types/genera were compared, indoor and outdoor Geotrichum and sterile mycelia of aerodynamic sizes 1.1–3.3 μm were found to be significantly elevated in high allergic prevalence CCCs. Indeed, average geometric mean diameters (D_{g, ave}) of indoor and outdoor culturable fungi were consistently smaller in CCCs with high prevalence of allergies than those with low prevalence. We found significant associations of higher fungal concentrations, especially those with smaller aerodynamic sizes in CCCs situated near parks. There were no differences in fungal levels between CCCs with respect to their dampness profile mainly due to high CCC ventilation rates. Since particle size is a factor that determines where a fungi particle deposits in the respiratory tract, this study provides useful information in the etiology of wheeze and rhinitis symptoms among the CCC attending children.     

Detection of residual organochlorine and organophosphorus pesticides in agricultural soil in Rio Verde region of San Luis Potosi,Mexico

Organochlorine pesticides were intensively used in Mexico from 1950 until their ban and restriction in 1991. However, the presence of these compounds is commonly reported in many regions of the country. The aim of the present study was to identify and quantify residual organochlorine and organophosphorus pesticides in agricultural soil in Rio Verde region, San Luis Potosi state, which has been identified as possibly polluted by pesticides. Composed samples from 24 zones covering an area of approximately 5,440 ha were analyzed. The most frequently found pesticides were p,p´-DDT followed by ,p,p´-DDE, heptachlor, endosulfan and γ-HCH whose frequency rates were 100, 91, 83 and 54%, respectively. The concentration of p,p´-DDT in the crops grown in these soils was in the following order: chili > maize > tomato > alfalfa. The results obtained in this study show that p,p´-DDT values are lower or similar to those found in other agricultural regions of Mexico. Methyl and ethyl parathion were the most frequent organophosphate pesticide detected in 100% and 62.5% of the samples with average concentrations of 25.20 and 47.48 μg kg^–1, respectively. More research is needed to establish the background levels of pesticides in agricultural soils and their potential ecological and human health effects in this region.     

Assessment of indoor air in Austrian apartments with and without visible mold growth

Fungal spores are transported across great distances in the outdoor air and are also regularly found indoors. Building conditions and behavior-related problems in apartments may lead to massive growth of mold within a very short period of time.The aim of this study was to evaluate whether the visible growth of mold indoors influences the concentration of fungal spores in the air as well as the variety of their species. Samples were collected from 66 households in Austria. For each sampling, the corresponding outdoor air was measured as reference value. The size of the visible mold growth was categorized in order to correlate the extent of mold growth with the concentration of airborne spores as well as the fungal genera. In order to determine fungal spore concentrations in the air, the one-stage MAS-100^® air sampler was used. Malt extract agar (MEA) and dichloran glycerol agar (DG18) plates were used as culture media. The total colony forming units (CFU) per m³ were determined. The fungi were identified from the isolated colonies.The results show that in apartments visibly affected by mold, the median values were significantly higher than those of apartments without visible mold growth. The extent of visible mold growth is significantly correlated with both concentration of fungal spores (p<0.001) as well as the predominance of Penicillium sp. and Aspergillus sp. (p<0.001) in indoor air. The total fungal concentration of Penicillium and Aspergillus in the air of apartments is recommended for assessing fungal exposure.     

Transport and attenuation of dissolved glyphosate and AMPA in a stormwater wetland

Glyphosate is an herbicide used widely and increasingly since the early 1990s in production of many crops and in urban areas. However, knowledge on the transport of glyphosate and its degradation to aminomethylphosphonic acid (AMPA) in ecosystems receiving urban or agricultural runoff is lacking. Here we show that transport and attenuation of runoff-associated glyphosate and AMPA in a stormwater wetland differ and largely vary over time. Dissolved concentrations and loads of glyphosate and AMPA in a wetland receiving runoff from a vineyard catchment were assessed during three consecutive seasons of glyphosate use (March to June 2009, 2010 and 2011). The load removal of glyphosate and AMPA by the wetland gradually varied yearly from 75% to 99%. However, glyphosate and AMPA were not detected in the wetland sediment, which emphasises that sorption on the wetland vegetation, which increased over time, and biodegradation were prevailing attenuation processes. The relative load of AMPA as a percentage of total glyphosate increased in the wetland and ranged from 0% to 100%, which indicates the variability of glyphosate degradation via the AMPA pathway. Our results demonstrate that transport and degradation of glyphosate in stormwater wetlands can largely change over time, mainly depending on the characteristics of the runoff event and the wetland vegetation. We anticipate our results to be a starting point for considering degradation products of runoff-associated pesticides during their transfer in wetlands, in particular when using stormwater wetlands as a management practice targeting pesticide attenuation.     

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.     

菌源对多环芳烃降解菌的筛选及降解性能的影响

高效降解菌的筛选对利用生物修复技术有效去除环境中的多环芳烃具有重要意义。分别以石油污染土壤和焦化废水活性污泥为菌源,分离出芘降解菌和混合PAHs(菲、荧蒽和芘)降解菌共14株并对其降解性能进行对比研究。结果表明,筛选得到的菌株分别属于9个菌属,其中2种菌源共有的菌属为Mycobacterium sp.、Ralstonia sp.和Shinella sp.。芘和PAHs的高效降解菌(CP16和CM32)均属于分支杆菌属(Mycobacterium),来源于焦化废水活性污泥;菌株CP16对芘(50mg/L)的7 d降解率为74.99%,CM32对PAHs(菲50 mg/L、荧蒽和芘各10 mg/L)的7 d降解率为100%。因此,以焦化废水活性污泥为菌源更有利于获得高效的多环芳烃降解菌。     

Characteristics,determinants, and spatial variations of ambient fungal levels in the subtropical Taipei metropolis

This study was conducted to investigate the temporal and spatial distributions, compositions, and determinants of ambient aeroallergens in Taipei, Taiwan, a subtropical metropolis. We monitored ambient culturable fungi in Shin-Jhuang City, an urban area, and Shi-Men Township, a rural area, in Taipei metropolis from 2003 to 2004. We collected ambient fungi in the last week of every month during the study period, using duplicate Burkard portable samplers and Malt Extract Agar. The median concentration of total fungi was 1339 colony-forming units m⁻³ of air over the study period. The most prevalent fungi were non-sporulating fungi, Cladosporium, Penicillium, Curvularia and Aspergillus at both sites. Airborne fungal concentrations and diversity of fungal species were generally higher in urban than in rural areas. Most fungal taxa had significant seasonal variations, with higher levels in summer. Multivariate analyses showed that the levels of ambient fungi were associated positively with temperature, but negatively with ozone and several other air pollutants. Relative humidity also had a significant non-linear relationship with ambient fungal levels. We concluded that the concentrations and the compositions of ambient fungi are diverse in urban and rural areas in the subtropical region. High ambient fungal levels were related to an urban environment and environmental conditions of high temperature and low ozone levels.     

New bacterial strain of the genus Ochrobactrum with glyphosate-degrading activity

Thirty bacterial strains with various abilities to utilize glyphosate as the sole phosphorus source were isolated from farm soils using the glyphosate enrichment cultivation technique. Among them, a strain showing a remarkable glyphosate-degrading activity was identified by biochemical features and 16S rRNA sequence analysis as Ochrobactrum sp. (GDOS). Herbicide (3 mM) degradation was induced by phosphate starvation, and was completed within 60 h. Aminomethylphosphonic acid was detected in the exhausted medium, suggesting glyphosate oxidoreductase as the enzyme responsible for herbicide breakdown. As it grew even in the presence of glyphosate concentrations as high as 200 mM, Ochrobactrum sp. could be used for bioremediation purposes and treatment of heavily contaminated soils.     

In vitro effects of various xenobiotics on Fusarium spp. strains isolated from cereals

This study aimed to determine the susceptibility of Fusarium spp. strains isolated from cereals to selected heavy metals, fungicides and silver nanoparticles. The experiments were conducted using 50 Fusarium strains belonging to five species: F. graminearum, F. culmorum, F. oxysporum, F. sporotrichioides and F. avenaceum. The strains were found to be highly resistant to Pb²⁺ and Zn²⁺. Medium resistance to Cu²⁺ and Mn²⁺ and low resistance to Cd²⁺ and Fe³⁺ was also observed. Among the tested fungicides, formulations containing azoxystrobin, prochloraz and tebuconazole proved to be the most effective in inhibiting the growth of fungi, as they affected fungal growth in each of the applied doses. Susceptibility of Fusarium spp. to nanosilver, demonstrated in this study, shows the legitimacy of using nanostructures as fungicidal agents. The results confirm high diversity of the analyzed fungal species in terms of susceptibility to the tested substances, and encourage to continue research on the resistance of Fusarium spp. to various fungicidal agents.     

Fungal levels in houses in the Fukushima Daiichi Nuclear Power Plant evacuation zone after the Great East Japan Earthquake

Residences located within 20 km of the damaged Fukushima Daiichi Nuclear Power Plant were evacuated shortly after the Great East Japan Earthquake. The levels of airborne and surface fungi were measured in six houses in the evacuation zone in August 2012 and February 2013. Airborne fungal levels in all of the houses in the summer were higher than the environmental standard levels for residential houses published in Architectural Institute of Japan (>1000 colony-forming units [CFU]/m³). In two houses whose residents rarely returned to visit, fungal levels were extremely high (>52,000 CFU/m³). Although fungal levels in the winter were much lower than those in the summer, they were still higher than environmental standard levels in several houses. Indoor fungal levels were significantly inversely related to the frequency with which residents returned, but they were not correlated with the air exchange rates, temperature, humidity, or radiation levels. Cladosporium spp. and Penicillium spp. were detected in every house. Aspergillus section Circumdati (Aspergillus ochraceus group) was also detected in several houses. These fungi produced ochratoxin A and ochratoxin B, which have nephrotoxic and carcinogenic potential. The present study suggests that further monitoring of fungal levels is necessary in houses in the Fukushima Daiichi Nuclear Power Plant evacuation zone, and that some houses may require fungal disinfection.

Implications: The results suggest that residents’ health could be at risk owing to the high levels of airborne fungi and toxic fungi Aspergillus section Circumdati. Therefore, monitoring and decontamination/disinfection of fungi are strongly recommended before residents are allowed to return permanently to their homes. In addition, returning to home with a certain frequency and adequate ventilation are necessary during similar situations, e.g., when residents cannot stay in their homes for a long period, because fungal levels in houses in the Fukushima Daiichi Nuclear Power Plant evacuation zone were inversely correlated with the frequency with which residents returned to visit their houses.     

Toxicity of pesticides toward human immune cells U-937 and HL-60

Abstract

The industrialization of the agricultural sector has significantly increased the use of chemicals such as pesticides. Therefore, exposure to them is unavoidable, which makes it necessary to assess their safety for humans at actual exposure doses. This paper aims to determine toxicity of three types of pesticides toward human immune cells (HL-60 and U-937): glyphosate (GLY), deltamethrin (DEL), and chlorothalonil (CHL). Cell viability, membrane integrity, inflammation induction, and antioxidant activity were evaluated to determine differences in cellular response to the tested plant protection agents. In experimental models, all tested substances caused increased mortality of cells after only 24?h. Cell membrane damage was recorded under DEL and CHL influences. The largest disintegration of the cell membrane was due to the action of 100?μg/mL DEL for U-937 and CHL at 1?μg/mL for HL-60. GLY at a concentration of 3,600?μg/mL caused significant peroxidation of U-937 cells’ lipids. CHL-induced inflammation in both types of cells tested. DEL and GLY also induced antioxidant activity in cells. These results lead to the conclusion that the tested pesticides act cytotoxically to the cells of the human immune system in doses to which both farmers and consumers are exposed.     

Potentiation of antifungal effect of a mixture of two antifungal fractions obtained from Baccharis glutinosa and Jacquinia macrocarpa plants

The aim of the present work was to evaluate the effect of mixtures of antifungal fractions extracted from Baccharis glutinosa and Jacquinia macrocarpa plants on the development of the filamentous fungi Aspergillus flavus and Fusarium verticillioides. The minimal inhibitory concentration that inhibited 50% of growth (MIC₅₀) of each plant antifungal fraction was determined from the percentage radial growth inhibition of both fungi. Binomial mixtures made with both plant fractions were used at their MIC₅₀ to determine the Fractional Inhibitory Concentration index (FIC index) for each fungus in order to evaluate their synergistic effect. Each synergistic mixture was analyzed in their effect on spore germination, spore size, spore viability, mitotic divisions, hyphal diameter and length, and number of septa per hypha. Some antifungal mixtures, even at low concentrations, showed higher antifungal effect than those of the individual antifungal fraction. The FIC indices of mixtures that showed the highest antifungal activity against A. flavus and F. verticillioides were 0.5272 and 0.4577, respectively, indicating a synergistic effect against both fungi. Only 12% and 8% of the spores of A. flavus and F. verticillioides, respectively, treated with the synergistic mixtures, were able to germinate, although their viability was not affected. An increase in the number of septa per hypha of both fungi was observed. The results indicated that the synergistic mixtures strongly affected the fungal growth even at lower concentrations than those of the individual plant fractions.     

DIFFERENTIAL EFFECTS OF THE HERBICIDES BENSULFURON AND CINOSULFURON ON SOIL MICROORGANISMS

Bensulfuron toxicity on soil microbes was evaluated by the methods used in a previous study on cinosulfuron; the effects of the two sulfonylureas were compared. Cinosulfuron and bensulfuron, at the normal field application rate and 100 times higher, had no effect on the total number of bacteria and nitrifiers, or on the respiration activity in the soil, but they did decrease the nitrification activity. In vitro toxicity tests carried out on representative soil microbial strains using bensulfuron at 50 mg l^?1 showed some inhibition of three of the 17 bacterial strains and strong inhibition of almost all the 12 fungal strains; cinosulfuron had had no effect on any of these strains in the previous study. It is concluded that, compared with cinosulfuron, bensulfuron is potentially more toxic on soil heterotrophic microorganisms, but only at very high concentrations that are nearly impossible to reach with the usual agricultural use of the herbicides. However, autotrophic nitrifiers were more sensitive to both sulfonylureas than the other microorganisms.