The effect of triclosan on microbial community structure in three soils

The application of sewage sludge to land can expose soils to a range of associated chemical toxicants. In this paper we explore the effects of the broad spectrum anti-microbial compound triclosan on the phenotypic composition of the microbial communities of three soils of contrasting texture (loamy sand, sandy loam and clay) using phospholipid fatty-acid (PLFA) analysis. Each soil type was dosed and subsequently re-dosed 6 weeks later with triclosan at five nominal concentrations in microcosms (10, 100, 500, 1000 mg kg⁻¹ and a zero-dose control). PLFA profiles were analysed using multivariate statistics focussing on changes in the soil phenotypic community structure. Additionally, ratios of fungal:bacterial PLFA indicators and cyclo:mono-unsaturated PLFAs (a common stress indicator) were calculated. It was hypothesised that triclosan addition would alter the community structure in each soil with a particular effect on the fungal:bacterial ratio, since bacteria are likely to be more susceptible to triclosan than fungi. It was also hypothesised that the PLFA response to re-dosing would be suppressed due to acclimation. Although the microbial community structure changed over the course of the experiment, the response was complex. Soil type and time emerged as the most important explanatory factors. Principal component analysis was used to detect phenotypic responses to different doses of triclosan in each soil. As expected, there was a significant increase in the fungal:bacterial ratio with triclosan dose especially in treatments with the highest nominal concentrations. Furthermore, the PLFA response to re-dosing was negligible in all soils confirming the acclimation hypothesis.     

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.     

Epiphytic lichen community dynamics in deciduous forests around a phosphorus fertiliser factory in Central Lithuania

The detailed dynamics of epiphytic lichen communities were observed while studying permanent quadrats in the zone of influence of a phosphorus fertiliser factory in central Lithuania. The most significant changes were induced by several factors: changes in macroenvironment (increase of illumination), bark scaling, succession processes, individual growth characteristics of the community members, and influence of fungal infection and invertebrate grazing. None of these changes could be directly linked with air pollution. These observations have shown that in conditions of more or less stable pollution, epiphytic community dynamics should be evaluated with care, the best indicators of the characteristics of the communities being species richness and presence/absence and abundance of indicator (nitrophilous or acidophilous) species.     

Fungicide impacts on microbial communities in soils with contrasting management histories

The impacts of the fungicides azoxystrobin, tebuconazole and chlorothalonil on microbial properties were investigated in soils with identical mineralogical composition, but possessing contrasting microbial populations and organic matter contents arising from different management histories. Degradation of all pesticides was fastest in the high OM/biomass soil, with tebuconazole the most persistent compound, and chlorothalonil the most readily degraded. Pesticide sorption distribution coefficient (K(d)) did not differ significantly between the soils. Chlorothalonil had the highest K(d) (97.3) but K(d) for azoxystrobin and tebuconazole were similar (13.9 and 12.4, respectively). None of the fungicides affected microbial biomass in either soil. However, all fungicides significantly reduced dehydrogenase activity to varying extents in the low OM/biomass soil, but not in the high OM/biomass soil. The mineralization of subsequent applications of herbicides, which represents a narrow niche soil process was generally reduced in both soils by azoxystrobin and chlorothalonil. 16S rRNA-PCR denaturing gradient gel electrophoresis (DGGE) indicated that none of the fungicides affected bacterial community structure. 18S rRNA PCR-DGGE analysis revealed that a small number of eukaryote bands were absent in certain fungicide treatments, with each band being specific to a single fungicide-soil combination. Sequencing indicated these represented protozoa and fungi. Impacts on the specific eukaryote DGGE bands showed no relationship to the extent to which pesticides impacted dehydrogenase or catabolism of herbicides.     

Effect of simulated acid rain on quantity of epiphytic microfungi on Scots pine (Pinus sylvestris L.) needles

The effects of simulated acid rain on the phyllosphere microflora of pine (Pinus sylvestris L.) were studied experimentally in northern Finland during the summer of 1988. Trees were irrigated with artificial acid rain of pH4 and pH3 (H(2)SO(4) and HNO(3), weight ratio of S:N=2.86:1). Untreated trees and trees irrigated with spring water (pH6) were used as controls. Two sampling heights (0.5m and 2m) were used. The needles were colonized exclusively by epiphytic fungi, mainly Aureobasidium pullulans (de Bary) Arnaud. The lower branches had significantly more epiphytic fungi than the upper branches. Compared to the control trees, the numbers of epiphytic fungi were significantly decreased on the needles of trees irrigated with acid rain. Acid rain affected the number of epiphytic fungi equally at both sampling heights. The species composition of the epiphytic fungi was not affected by the acid treatments.     

A comparative analysis of endophytic bacterial communities associated with hyperaccumulators growing in mine soils

Interactions between endophytic bacterial communities and hyperaccumulators in heavy metal-polluted sites are not fully understood. In this study, the diversity of stem-associated endophytic bacterial communities of two hyperaccumulators (Solanum nigrum L. and Phytolacca acinosa Roxb.) growing in mine soils was investigated using molecular-based methods. The denaturing gradient gel electrophoresis (DGGE) analysis showed that the endophytic bacterial community structures were affected by both the level of heavy metal pollution and the plant species. Heavy metal in contaminated soil determined, to a large extent, the composition of the different endophytic bacterial communities in S. nigrum growing across soil series (five sampling spots, and the concentration of Cd is from 0.2 to 35.5 mg/kg). Detailed analysis of endophytic bacterial populations by cloning of 16S rRNA genes amplified from the stems of the two plants at the same site revealed a different composition. A total of 51 taxa at the genus level that included α-, β-, and γ-Proteobacteria (68.8% of the two libraries clones), Bacteroidetes (9.0% of the two libraries clones), Firmicutes (2.0% of the two libraries clones), Actinobacteria (16.4% of the two libraries clones), and unclassified bacteria (3.8% of the two libraries clones) were found in the two clone libraries. The most abundant genus in S. nigrum was Sphingomonas (23.35%), while Pseudomonas prevailed in P. acinosa (21.40%). These results suggest that both heavy metal pollution and plant species contribute to the shaping of the dynamic endophytic bacterial communities associated with stems of hyperaccumulators.     

Effect of moisture and compost on fate of azoxystrobin in soils

The effect of compost-amendment and moisture status on the persistence of azoxystrobin [methyl (E)-2-{2-(6-(2-cyanophenoxy) pyrimidin-4-yloxy) phenyl}-3-methoxyacrylate], a strobilurin fungicide, in two rice-growing soils was studied. Azoxystrobin is more sorbed in the silt loam (K _f – 4.66) soil than the sandy loam (K _f – 2.98) soil. Compost-amendment at 5 % levels further enhanced the azoxystrobin sorption and the respective K _f values in silt loam and sandy loam soils were 8.48 and 7.6. Azoxystrobin was more persistent in the sandy loam soil than the silt loam soil. The half–life values of azoxystrobin in nonflooded and flooded silt loam soil were 54.7 and 46.3 days, respectively. The corresponding half–life values in the sandy loam soils were 64 and 62.7 days, respectively. Compost application enhanced persistence of azoxystrobin in the silt loam soil under both moisture regimes and half-life values in non–flooded and flooded soils were 115.7 and 52.8 days, respectively. However, compost enhanced azoxystrobin degradation in the sandy loam soil and half-life values were 59 (nonflooded) and 54.7 days (flooded). The study indicates that compost amendment enhanced azoxystrobin sorption in the soils. Azoxystrobin is more persistent in non-flooded soils than the flooded soils. Compost applications to soils had mixed effect on the azoxystrobin degradation.     

Leaching and sorption of neonicotinoid insecticides and fungicides from seed coatings

Seed coatings are a treatment used on a variety of crops to improve production and offer protection against pests and fungal outbreaks. The leaching of the active ingredients associated with the seed coatings and the sorption to soil was evaluated under laboratory conditions using commercially available corn and soybean seeds to study the fate and transport of these pesticides under controlled conditions. The active ingredients (AI) included one neonicotinoid insecticide (thiamethoxam) and five fungicides (azoxystrobin, fludioxonil, metalaxyl, sedaxane thiabendazole). An aqueous leaching experiment was conducted with treated corn and soybean seeds. Leaching potential was a function of solubility and seed type. The leaching of fludioxonil, was dependent on seed type with a shorter time to equilibrium on the corn compared to the soybean seeds. Sorption experiments with the treated seeds and a solution of the AIs were conducted using three different soil types. Sorption behavior was a function of soil organic matter as well as seed type. For most AIs, a negative relationship was observed between the aqueous concentration and the log K_oc. Sorption to all soils tested was limited for the hydrophilic pesticides thiamethoxam and metalaxyl. However, partitioning for the more hydrophobic fungicides was dependent on both seed type and soil properties. The mobility of fludioxonil in the sorption experiment varied by seed type indicating that the adjuvants associated with the seed coating could potentially play a role in the environmental fate of fludioxonil. This is the first study to assess, under laboratory conditions, the fate of pesticides associated with seed coatings using commercially available treated seeds. This information can be used to understand how alterations in agricultural practices (e.g., increasing use of seed treatments) can impact the exposure (concentration and duration) and potential effects of these chemicals to aquatic and terrestrial organisms.     

Effects of various ozone exposures on the susceptibility of bean leaves (Phaseolus vulgaris L.) to Botrytis cinerea

The effects of various ozone exposures in predisposing bean leaves (Phaseolus vulgaris L.) to Botrytis cinerea have been investigated under laboratory conditions. Seedlings of two bean cultivars were exposed to incremental ozone concentrations (120, 180 and 270 microg m(-3) for 8-h day(-1)) for five days and primary leaves were subsequently inoculated with conidia suspended in water or in an inorganic phosphate solution (Pi), and with mycelium. Ozone injury increased with increasing ozone concentration and was much higher in the ozone-sensitive cultivar 'Pros' than in the ozone-insensitive 'Groffy'. Ozone only increased the number of lesions on leaves of Pros after inoculation with either of the conidial suspensions. The Pi-stimulated infection in Groffy was reduced by the lower ozone concentrations. Ozone decreased lesion expansion after inoculation with mycelium. In a chronic fumigation experiment, plants of the two cultivars were exposed to 90 microg m(-3) (7-h day(-1)) and the primary and the oldest tree trifoliate leaves were inoculated after five and seven weeks of exposure. Ozone enhanced the senescence-related injury only in Pros. The number of lesions was not influenced by ozone for either cultivar, conidial suspension or inoculation date. Lesion expansion after inoculation with mycelium was generally reduced in exposed plants. Thus, contrasting effects of ozone on the susceptibility of bean leaves to B. cinerea were observed depending on the cultivar, the conidial suspension, the disease parameter and the ozone exposure pattern. In extrapolating the laboratory results to the field, it is suggested that episodic and chronic exposures to ambient ozone are of minor importance in increasing the susceptibility of bean leaves to B. cinerea.     

Adsorption, transformation, and bioavailability of the fungicides carbendazim and iprodione in soil, alone and in combination

When studying the effect of mixtures of toxic substances on soil organisms, attention must be paid to peculiarities in exposure to mixtures as opposed to that of single toxicants. The fungicides carbendazim and iprodione compete in the adsorption to soil. The presence of iprodione reduced the adsorption of carbendazim by 30%, while carbendazim reduced the adsorption of iprodione by 70%. Iprodione had little effect on the transformation rate of carbendazim in soil. However, carbendazim retarded the transformation of iprodione in soil by 26%. The concentration of the fungicides in pore water was found to be substantially higher for mixtures than when a fungicide alone was present in the soil. The effect of the additional fungicide on the concentration is especially apparent in the period following the first 1 to 2 weeks of the incubation. The inclusion of copper in the mixture has little additional effect on the concentration of the fungicides in pore water.     

Effects of fungicides triadimefon and propiconazole on soil bacterial communities

The impact of fungicides triadimefon and propiconazole on soil bacterial populations from a strawberry field was investigated. Two fungicides were applied to the soil at concentrations of 10 mg/kg or 100 mg/kg with soil water contents 20.2% (fresh soil water content) or 26.0% (field capacity). Changes in bacterial communities were assessed using DNA extraction, polymerase chain reaction (PCR) amplification of the 16S rDNA and denaturing gradient gel electrophoresis (DGGE). High performance liquid chromatography (HPLC) was utilized to detect the residue of fungicides in soils. The results showed that propiconazole was more persistent than triadimefon in soils, and the two soil water contents did not cause significant differences in dissipation rates between the two fungicides. A high concentration of propiconazole could inhibit the existence of soil microbes while one of triadimefon might induce the microbial population in the first stage. From unweighted pair-group method using arithmetic averages (UPGMA) dendrograms, the effect of triadimefon and propiconazole at the two applied concentrations on a soil bacterial community could be long term. After triadimefon was applied for 60 days and propiconazole for 75 days, the compositions of microbial communities were not recovered. From the viewpoint of environmental protection, it was of significant importance to pay more attention not only to the residues of pesticide but also to the change in soil microbial communities.     

Direct Effects of Atmospheric Sulfate Deposition on Vegetation

Acid sulfate aerosol (500 μg/m³) had no effect on soybean or pinto bean after a single 4-h exposure. However, visible Injury and chlorophyll loss occurred when plants were sequentially exposed to acid aerosol and ozone (380 μg/m³) for 4 h. In yellow poplar seedlings exposed to ozone (200 μg/m³), sulfur dioxide (210 μg/m³) and simulated rain solutions (pH 5.6, 4.3 and 3.0) for 6 weeks, root dry weight, leaf area increase, mean relative growth rate and unit leaf rate decreased linearly with pH in ozone-treated plants. However, unit leaf rate and mean relative growth rate increased linearly in response to sulfur dioxide as solution acidity increased. Ambient wet and dry sulfate concentrations appear insufficient to directly impact vegetation.     

Microbial community structure in a silty clay loam soil after fumigation with three broad spectrum fungicides

The short-term effect of three broad spectrum fungicides on microbial activity, microbial biomass, soil ergosterol content, and phospholipid fatty acid (PLFA) profiles was studied. A silty clay loam soil was treated separately with captan, chlorothalonil and carbendazim at three different dosages of each fungicide. Chlorothalonil and carbendazim significantly altered soil microbial activity. However, changes in soil microbial biomass were only observed in soil treated with higher dosages of these fungicides. All dosages of fungicides significantly decreased fungal biomass as estimated by soil ergosterol content. PLFA analysis indicated that there was a shift in PLFA pattern. Higher dosages of all three fungicides decreased a straight-chain PLFA 22:0. In addition, soil treated with carbendazim increased cyclopropyl fatty acids. Compared to untreated soil, higher dosages of both captan and chlorothalonil affected PLFA 10Me 16:0, indicating that these fungicides can reduce actinomycetes population. Finally, our results suggest that application of both captan and chlorothalonil decreased Gram-positive to Gram-negative ratio.     

The dissipation of three fungicides in a biobed organic substrate and their impact on the structure and activity of the microbial community

Biopurification systems (BPS) have been introduced to minimise the risk for point source contamination of natural water resources by pesticides. Their depuration efficiency relies mostly on the high biodegradation of their packing substrate (biomixture). Despite that, little is known regarding the interactions between biomixture microflora and pesticides, especially fungicides which are expected to have a higher impact on the microbial community. This study reports the dissipation of the fungicides azoxystrobin (AZX), fludioxonil (FL) and penconazole (PC), commonly used in vineyards, in a biomixture composed of pruning residues and straw used in vineyard BPS. The impact of fungicides on the microbial community was also studied via microbial biomass carbon, basal respiration and phospholipid fatty acid analysis. AZX dissipated faster (t _1/2?=?30.1 days) than PC (t _1/2?=?99.0 days) and FL (t _1/2?=?115.5 days). Fungicides differently affected the microbial community. PC showed the highest adverse effect on both the size and the activity of the biomixture microflora. A significant change in the structure of the microbial community was noted for PC and FL, and it was attributed to a rapid inhibition of the fungal fraction while bacteria showed a delayed response which was attributed to indirect effects by the late proliferation of fungi. All effects observed were transitory and a full recovery of microbial indices was observed 60 days post-application. Overall, no clear link between pesticide persistence and microbial responses was observed stressing the complex nature of interactions between pesticides in microflora in BPS.     

Retention capacity of an organic bio-mixture against different mixtures of fungicides used in vineyards

A laboratory experiment was carried out to test the efficiency of a bio-mixture made up of pruning residues at two (PR2) and five (PR5) years of composting and wheat straw (STW) in the biological cleaning of water contaminated by different mixtures of fungicides usually employed in vineyards. The experiment was conducted and reproduced at a scale of 1:100 of operating field conditions. Commercial formulates of penconazole (PC), (RS)-1-[2-(2,4-dichlorophenyl)pentyl]-1H-1,2,4-triazole), dimetomorph (DM), (EZ)-4-[3-(4-chlorophenyl)-3-(3,4-dimethoxyphenyl)acryloyl]morpholine), azoxystrobin (AZ), (methyl (E)-2-{2-[6-(2-cyanophenoxy)pyrimidin-4-yloxy]phenyl}-3-methoxyacrylate), iprovalicarb (IP), (isopropyl 2-methyl-1-[(RS)-1-p-tolylethyl]carbamoyl-(S)-propylcarbamate), metalaxyl (MX), (methyl N-(methoxyacetyl)-N-(2,6-xylyl)-DL-alaninate), fludioxonil (FL), (4-(2,2-difluoro-1,3-benzodioxol-4-yl)-1H-pyrrole-3-carbonitrile) and cyprodinil (CY), (4-cyclopropyl-6-methyl-N-phenylpyrimidin-2-amine) were mixed in water and discharged into the bio-mixture following the time schedule of the treatments carried out in the grapevine in real field conditions. At each treatment, contaminated water with fungicides was circulated repeatedly through the bio-mixture to enhance the sorption of fungicides. In fact, it retained them between 98–100% with the exception of MX of which it was able to retain only 90.5%. The adsorption/desorption experiment showed that repeated circulation of water, instead of enhancing MX retention, can easily remove about 30% of MX already adsorbed by the bio-mixture. This finding suggests that water contaminated by very mobile pesticides should be discharged at the end of field treatments without re-circulating the water in order to avoid the release of pesticides weakly adsorbed on the bio-mixture.     

Effect of Dursban 480 EC (chlorpyrifos) and Talstar 10 EC (bifenthrin) on the physiological and genetic diversity of microorganisms in soil

This investigation was undertaken to determine the impact of the insecticides Dursban 480 EC (with organophosphate compound chlorpyrifos as the active ingredient) and Talstar 10 EC (with pyrethroid bifenthrin as the active ingredient) on the respiration activity and microbial diversity in a sandy loam luvisol soil. The insecticides were applied in two doses: the maximum recommended dose for field application (15 mg kg^?1 for Dursban 480 EC and 6 mg kg^?1 for Talstar 10 EC) and a 100-fold higher dose for extrapolation of their effect. Bacterial and fungal genetic diversity was analysed in soil samples using PCR DGGE and the functional diversity (catabolic potential) was studied using BIOLOG EcoPlates at 1, 3, 7, 14, 28, 56 and 112 days after insecticide application. Five bacterial groups (α, β, γ proteobacteria, firmibacteria and actinomycetes) and five groups of fungi or fungus-like microorganisms (Ascomycota, Basidiomycota, Chytridiomycota, Oomycota and Zygomycota) were analysed using specific primer sets. This approach provides high resolution of the analysis covering majority of microorganisms in the soil. Only the high-dose Dursban 480 EC significantly changed the community of microorganisms. We observed its negative effect on α- and γ-proteobacteria, as the number of OTUs (operational taxonomic units) decreased until the end of incubation. In the β-proteobacteria group, initial increase of OTUs was followed by strong decrease. Diversity in the firmibacteria, actinomycetes and Zygomycota groups was minimally disturbed by the insecticide application. Dursban 480 EC, however, both positively and negatively affected certain species. Among negatively affected species Sphingomonas, Flavobacterium or Penicillium were detected, but Achromobacter, Luteibacter or Aspergillus were supported by applied insecticide. The analysis of BIOLOG plates using AWCD values indicated a significant increase in metabolic potential of microorganisms in the soil after the high-dose Dursban application. Analysis of respiration demonstrated high microbial activity after insecticide treatments; thus, microbial degradation was relatively fast. The half-life of the active insecticide compounds were estimated within the range of 25 to 27 days for Talstar and 6 to 11 days for Dursban and higher doses stimulated degradation. The recommended dose levels of both insecticides can be considered as safe for microbial community in the soil.     

Shaping effects of rice,wheat, maize,and soybean seedlings on their rhizosphere microbial community

The rhizosphere microbiome plays critical roles in plant growth and is an important interface for resource exchange between plants and the soil environment. Crops at various growing stages, especially the seedling stage, have strong shaping effects on the rhizosphere microbial community, and such community reconstruction will positively feed back to the plant growth. In the present study, we analyzed the variations of bacterial and fungal communities in the rhizosphere of four crop species: rice, soybean, maize, and wheat during successive cultivations (three repeats for the seedling stages) using 16S rRNA gene and internal transcribed spacer (ITS) high-throughput sequencing. We found that the relative abundances of specific microorganisms decreased after different cultivation times, e.g., Sphingomonas, Pseudomonas, Rhodanobacter, and Caulobacter, which have been reported as plant-growth beneficial bacteria. The relative abundances of potential plant pathogenic fungi Myrothecium and Ascochyta increased with the successive cultivation times. The co-occurrence network analysis showed that the bacterial and fungal communities under maize were much more stable than those under rice, soybean, and wheat. The present study explored the characteristics of bacteria and fungi in crop seedling rhizosphere and indicated that the characteristics of indigenous soil flora might determine the plant growth status. Further study will focus on the use of the critical microorganisms to control the growth and yield of specific crops.

     

Residues behavior of some fungicides applied on two greenhouse tomato varieties different in shape and weight

The degradation of 12 fungicides (azoxystrobin, cymoxanil, cyproconazole, cyprodinil, fenarimol, fludioxonil, iprovalicarb, mepanipyrim, penconazole, pyrimethanil, tolclofos-methyl, triadimenol), commonly used in pest management strategies on Sardinian greenhouse tomato crops was studied. A different residue behaviour was observed between the studied cultivar. On the smaller, a "cherry" type tomato, field data showed an initial residue mostly higher than the "beefsteak" tomato. In any case, except for penconazole, all pesticide residues were below their maximum residue levels (MRLs) reaching the pre-harvest interval (PHI). On both cultivar, triadimenol and cymoxanil residues completely disappeared reaching their PHI, while iprovalicarb, fenarimol, and fludioxonil disappeared in a time of 17 to 24 days. On the contrary, azoxystrobin, cyproconazole, cyprodinil, penconazole, tolclofos-methyl, mepanipyrim, and pyrimethanil showed a long persistence on both tested cultivar and may have residual problems due to an accumulation effect if repeated field treatments will be performed. The first group of molecules according to their rapid degradation could be used in low pesticide-input management in order to obtain tomatoes with low or no detectable residues.     

Structural changes in soil communities after triclopyr application in soils invaded by Acacia dealbata Link

Triclopyr is a commonly used herbicide in the control of woody plants and can exhibit toxic effects to soil microorganisms. However, the impact on soils invaded by plant exotics has not yet been addressed. Here, we present the results of an 18-month field study conducted to evaluate the impact of triclopyr on the structure of fungal and bacterial communities in soils invaded by Acacia dealbata Link, through the use of denature gradient gel electrophoresis. After triclopyr application, analyses of bacterial fingerprints suggested a change in the structure of the soil bacterial community, whereas the structure of the soil fungal community remained unaltered. Bacterial density and F:B ratio values changed across the year but were not altered due to herbicide spraying. On the contrary, fungal diversity was increased in plots sprayed with triclopyr 5 months after the first application. Richness and diversity (H´) of both bacteria and fungi were not modified after triclopyr application.     

The biochemical activity of soil contaminated with fungicides

Excess fungicides can pose a serious threat to the soil environment. Fungicides can lower the microbiological and biochemical activity of soil and lead to yield declines. Soils contaminated with fungicides have to be remediated to maintain the optimal function of soil ecosystems. This study evaluates the effect of neutralizing substances on soil enzymatic activity and the yield of Triticum aestivum L. in soil contaminated with fungicides. Sandy loam (Eutric Cambisols) with pH_KCl 7.0 was contaminated with an aqueous solution of Amistar 250 SC and Falcon 460 EC in the following doses: 0 (soil without fungicide – treated as a control), RD (dose recommended by the manufacturer) and 300?×?RD (dose 300-fold higher than the recommended dose). Soil was supplemented with bentonite and basalt meal at a dose of 10?g kg^?1 DM of soil (dry mass of soil). The fungicide dose recommended by the manufacturer did not induce changes in soil enzymatic activity or the yield of T. aestivum L. Our findings indicate that the tested fungicides can be safely applied to protect crops against fungal pathogens. However, when applied at the dose of 300?×?RD, the tested fungicides strongly inhibited soil enzymatic activity and disrupted the growth and development of spring wheat. Soil supplementation with bentonite and basalt meal improved the yield of T. aestivum L., and bentonite was more effective in reducing fungicide stress. The analyzed substances were not highly effective in restoring biochemical homeostasis in soil.