In vitro toxicity of selected fungicides from the groups of benzimidazoles and demethylation inhibitors to Cladobotryum dendroides and Agaricus bisporus

Twenty microfungal isolates were collected from diseased fruiting bodies of Agaricus bisporus sampled from Serbian mushroom farms during 2003–2007. Based on morphological characteristics and pathogenicity tests, the isolates were identified as Cladobotryum dendroides. The isolates of C. dendroides and A. bisporusF56 and U3 were tested for sensitivity to several selected fungicides in vitro. C. dendroides isolates were found to be more sensitive to prochloraz manganese and flusilazole + carbendazim than to the other fungicides tested (EC₅₀ values were 0.09 and 0.11 mg L^{? 1}, respectively) and weakly resistant to thiophanate-methyl (EC₅₀ values ranged between 6.53 and 12.09 mg L^{? 1}). Selectivity indexes of the tested fungicides on both C. dendroidesand A. bisporusindicated that thiophanate-methyl, cyproconazole + carbendazim and flusilazole + carbendazim had much less selective fungitoxicity than benomyl, carbendazim and prochloraz manganese.     

Fungicide sensitivity of Trichoderma spp. from Agaricus bisporus farms in Serbia

Trichoderma species, the causal agents of green mould disease, induce great losses in Agaricus bisporus farms. Fungicides are widely used to control mushroom diseases although green mould control is encumbered with difficulties. The aims of this study were, therefore, to research in vitro toxicity of several commercial fungicides to Trichoderma isolates originating from Serbian and Bosnia-Herzegovina farms, and to evaluate the effects of pH and light on their growth. The majority of isolates demonstrated optimal growth at pH 5.0, and the rest at pH 6.0. A few isolates also grew well at pH 7. The weakest mycelial growth was noted at pH 8.0–9.0. Generally, light had an inhibitory effect on the growth of tested isolates. The isolates showed the highest susceptibility to chlorothalonil and carbendazim (ED₅₀ less than 1 mg L^?1), and were less sensitive to iprodione (ED₅₀ ranged 0.84–6.72 mg L^?1), weakly resistant to thiophanate-methyl (ED₅₀ = 3.75–24.13 mg L^?1), and resistant to trifloxystrobin (ED₅₀ = 10.25–178.23 mg L^?1). Considering the toxicity of fungicides to A. bisporus, carbendazim showed the best selective toxicity (0.02), iprodione and chlorothalonil moderate (0.16), and thiophanate-methyl the lowest (1.24), while trifloxystrobin toxicity to A. bisporus was not tested because of its inefficiency against Trichoderma isolates.     

In vitro toxicity of fungicides with different modes of action to alfalfa anthracnose fungus,Colletotrichum destructivum

Abstract

Sensitivity of 24 isolates of Colletotrichum destructivum O’Gara, collected from alfalfa plants in Serbia, to eight selected fungicides, was investigated in this study. Molecular identification and pathogenicity test of isolates tested were also performed. Fungicide sensitivity was evaluated in vitro, using mycelial growth assay method. All isolates exhibited significant pathogenicity, causing necrosis at the alfalfa seedling root tips two days after inoculation. Using the primer pair GSF1-SR1 and by comparing the amplified fragments of the tested isolates with the marker (M), the presence of the amplicon of the expected size of about 900?bp was determined for all isolates. The isolates tested in this study showed different sensitivity towards fungicides in vitro. Mycelial growth was highly inhibited by QoI (quinone outside inhibitors) fungicide pyraclostrobin (mean EC₅₀=0.39?µg mL^?1) and by DMI (demethylation-inhibiting) fungicide tebuconazole (mean EC₅₀=0.61?µg mL^?1), followed by azoxystrobin (mean EC₅₀=2.83?µg mL^?1) and flutriafol (mean EC₅₀=2.11?µg mL^?1). Multi-site fungicide chlorothalonil and MBC (methyl benzimidazole carbamate) fungicide thiophanate-methyl evinced moderate inhibition with mean EC₅₀=35.31 and 62.83?µg mL^?1, respectively. Thirteen isolates were sensitive to SDHI (succinate dehydrogenase inhibitors) fungicide boscalid and fluxapyroxad, (mean EC₅₀=0.49 and 0.19?µg mL^?1, respectively), while the rest of isolates were highly resistant.     

Remediation capacity of Cd and Pb ions by mycelia of Imleria badia,Laetiporus sulphureus,and Agaricus bisporus in vitro cultures

The goal of this study was to evaluate cadmium and lead accumulation ability of in vitro cultures biomass containing selected edible mushroom species derived from the environment (Laetiporus sulphureus, Imleria badia) and those of commercial origin (Agaricus bisporus). Atomic absorption spectrometry was used to evaluate the content of Cd(II) and Pb(II) on the medium supplemented with Cd(II) or Pb(II), each of them at the same concentration of 5·10^?5 M. The highest concentration of Cd(II) ions was determined in the biomass from L. sulphureus in vitro cultures, while the highest concentration of Pb(II) ions was found in the biomass from A. bisporus in vitro cultures. The greatest Cd(II) and Pb(II) accumulation ability in mycelium per dry weight was shown for L. sulphureus. Among the test species, biomass of A. bisporus showed the lowest ability for the bioaccumulation of Cd(II); however, comparable ability for the remediation of Pb(II) was provided by the biomasses from A. bisporus and I. badia in vitro cultures. The results confirm the possibility of using these mushroom species for remediation and indicate the relationship between bioaccumulation of heavy metals and the test species.     

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.     

Impact of three different fungicides on fungal epi- and endophytic communities of common bean (Phaseolus vulgaris) and broad bean (Vicia faba)

In this study, the impacts of three different fungicides to fungal phyllosphere communities on broad bean (Vicia faba, Fabaceae) and common bean (Phaseolus vulgaris, Fabaceae) were analyzed. The fungicides included copper, sulfur, and azoxystrobin. The plants were sowed, grown, and treated under conditions occurring in conventional and organic farming. A culture-based approach was used to identify changes in the phyllosphere fungal community after the treatment. Different effects on species richness and growth index of the epiphytic and endophytic communities for common bean and broad bean could be shown. Treatments with sulfur showed the weakest effect, followed by those based on copper and the systemic azoxystrobin, which showed the strongest effect especially on endophytic communities. The epiphytic fungal community took five weeks to recover after treatment with azoxystrobin. However, the effect of azoxystrobin on the endophytic community lasted more than five weeks. Finally, the data suggest that the surface structure of the host leaves have a huge impact on the mode of action that the fungicides exert.     

Occurrence of mycotoxins in wheat grains exposed to fungicides on fusarium head blight control in southern Brazil

Mycotoxins occurrence in wheat grains impose risks to human and animal health. The southern Brazil has favorable weather conditions for Fusarium graminearum infections and consequently for mycotoxins accumulation on grains. The goal of this study was to evaluate the behavior of new wheat commercial genotypes to Fusarium Head Blight (FHB), to control performance of new fungicide formulations and their relationship with mycotoxins concentration in grains. The manly mycotoxin occurrence on wheat grains in southern Brazil was deoxynivalenol (DON). Two cultivars showed high DON concentration above the tolerance limits (>3000 μg kg^?1). Many other mycotoxins monitored presented concentrations below method detection limit. Satisfactory levels of fungicide effectiveness were achieved against F. graminearum. Some fungicides promoted a satisfactory decrease on DON accumulation in grains. The best results were obtained when prothioconazole was present. SDHI (Succinate dehydrogenase inhibitors) + QoI (Quinone outside inhibitors) fungicides showed benefic effects at FHB control at field, but it did not promote satisfactory reduction on DON contamination. Fungicides can be used satisfactory for FHB control and reduce DON contamination in grains in southern Brazil. The presence of prothioconazole should be recommended. Some genotypes showed high DON concentration and it was not directly related with FHB severity at field.     

Metabolic responses of Eisenia fetida after sub-lethal exposure to organic contaminants with different toxic modes of action

Nuclear magnetic resonance (NMR) - based metabolomics has the potential to identify toxic responses of contaminants within a mixture in contaminated soil. This study evaluated the metabolic response of Eisenia fetida after exposure to an array of organic compounds to determine whether contaminant-specific responses could be identified. The compounds investigated in contact tests included: two pesticides (carbaryl and chlorpyrifos), three pharmaceuticals (carbamazephine, estrone and caffeine), two persistent organohalogens (Aroclor 1254 and PBDE 209) and two industrial compounds (nonylphenol and dimethyl phthalate). Control and contaminant-exposed metabolic profiles were distinguished using principal component analysis and potential contaminant-specific biomarkers of exposure were found for several contaminants. These results suggest that NMR-based metabolomics offers considerable promise for differentiating between the different toxic modes of action (MOA) associated with sub-lethal toxicity to earthworms.     

Ovarian activity of Drosophila melanogaster meigen (diptera), during a chronic intoxication with four fungicides: Anatomical and cytological study

Abstract

Two types of reactions were observed on the alteration of Drosophila vitellogenesis by the four fungicides used in this study. Dithane®M45 resulted in stimulation associated with egg retention. However, the other three fungicides (Benlate®, Bouillie bordelaise® and Euparène®) resulted in inhibition to a varying degree. Although the inhibition was comparatively limited due to Benlate® it induced an egg retention. The inhibition was very high due to Bouillie bordelaise® and Euparène®. With Bouillie bordelaise® an egg retention occured together with the reduction of vitellogenesis and caused an increase in the rate of the follicle resorption. The latter depended on the duration of treatment. With Euparène®, no egg retention was observed and the toxicity was only noticed on vitellogenesis.     

Toxicity of mancozeb,chlorothalonil, captan,fluopyram, boscalid,and difenoconazole to Didymella applanata isolates from Serbia

Field isolates of Didymella applanata, the causal agent of spur blight of raspberry, were evaluated in vitro for their sensitivity to mancozeb, chlorothalonil, captan, fluopyram, boscalid and difenoconazole. A total of 10 isolates, collected during 2013 at five localities in the major raspberry growing region in Serbia, and characterized as copper hydroxide, dithianon, and tebuconazole (sensitive), pyraclostrobin (sensitive or highly resistant) and fluazinam (sensitive or moderately resistant), were used in this study. The EC₅₀ values for the isolates ranged from 1.33 to 2.88 mg L^?1 for mancozeb, from 3.18 to 6.65 mg L^?1 for chlorothalonil, from 15.75 to 24.69 mg L^?1 for captan and from 1.80 to 8.20 mg L^?1 for fluopyram. The narrowest range of EC₅₀ values was recorded for difenoconazole (0.23–0.49 mg L^?1), whereas the widest range was obtained for boscalid (4.49–49.25 mg L^?1). The calculated resistance factors showed that all D. applanata isolates were sensitive to mancozeb, chlorothalonil, captan, and difenoconazole. Four isolates were moderately resistant to boscalid, while three of them were also moderately resistant to fluopyram. This finding of moderately resistant isolates to these SDHI fungicides indicates a possible cross-resistance which should be clarified in further investigations.     

Responses of different fungal and plant species to acetolactate synthase inhibitors and their derivatives

Abstract

The effect of sulfonylurea (SU) acetolactate synthases (ALS) inhibitors (chlorsulfuron and triasulfuron) and their 13 guanidine and 7 azole derivatives was studied on 17 fungal and 21 plant species. The efficacy of chlorsulfuron and triasulfuron against Fusarium oxysporum and Thielaviopsis basicola was higher than carbendazim. Two azole derivatives were highly selective for maize. The antifungal activity spectrum of both guanidine and azole derivatives differed from that of chlorsulfuron and triasulfuron, and did not exert notable antifungal activity in vitro. However, they had significant anti‐Fusarium efficacy on rye that surpassed that of carbendazim.     

Impact of a native Streptomyces flavovirens from mushroom compost on green mold control and yield of Agaricus bisporus

ABSTRACT

Thirty-five actinobacterial isolates, obtained from button mushroom (Agaricus bisporus) substrates (i.e., compost in different phases of composting, black peat or casing layer) in Serbia in 2014–2016 were tested in vitro against the causal agents of green mold in cultivated mushroom. Out of six most promising isolates, A06 induced 42.4% in vitro growth inhibition of Trichoderma harzianum T54, and 27.6% inhibition of T. aggressivum f. europaeum T77. The novel strain A06 was identified as Streptomyces flavovirens based on macroscopic and cultural characteristics and 16S rDNA sequence and used in mushroom growing room experiments. Actinobacteria had no negative influence on mycelial growth of the cultivated mushroom in compost in situ. Isolate S. flavovirens A06 enhanced mushroom yield significantly, up to 31.5%. The A06 isolate was more efficient in enhancing yield after inoculation with the compost mold T. aggressivum (26.1%), compared to casing mold T. harzianum (8%). Considering disease incidence, actinobacteria significantly prevented green mold in compost caused by T. aggressivum (6.8%). However, fungicide prochloraz-Mn had a more significant role in reducing symptoms of casing mold, T. harzianum, in comparison with actinobacteria (24.2 and 11.8%, respectively). No significant differences between efficacies of S. flavovirens A06 and the fungicide prochloraz-Mn against T. aggressivum were revealed. These results imply that S. flavovirens A06 can be used to increase mushroom yield and contribute to disease control against the aggressive compost green mold disease caused by Trichoderma aggressivum.     

Sensitivity of the zebrafish (Danio rerio) early life stage test for compounds with different modes of action

The sensitivity of the early life stage (ELS) toxicity test for two compounds with different modes of action was determined, and related to other toxicity tests with the same compounds. The zebrafish. Danio rerio, was used as a test organism, and the two model compounds were 1,2,3-trichlorobenzene (123TCB), a non-polar narcotic, and parathion, an acetylcholinesterase (AChE) inhibitor. Hatching and survival after 28 days were significantly reduced in the highest 123TCB treatment (263 microg/l), but not in any of the parathion treatments. Growth of the larvae was negatively affected at parathion concentrations above 20 microg/l, while AChE was only significantly inhibited at the highest concentration, 93 microg/l. No effects on growth were found in the 123TCB treatments. In comparison with acute and chronic studies with both compounds, the ELS test turned out to be less sensitive than chronic studies and more sensitive than acute studies. The difference in sensitivity between the tests systems seems however, to depend on the mode of action of the compound.     

Toxicity of twenty-two plant essential oils against pathogenic bacteria of vegetables and mushrooms

Toxicity of twenty-two essential oils to three bacterial pathogens in different horticultural systems: Xanthomonas campestris pv. phaseoli (causing blight of bean), Clavibacter michiganensis subsp. michiganensis (bacterial wilt and canker of tomato), and Pseudomonas tolaasii (causal agent of bacterial brown blotch on cultivated mushrooms) was tested. Control of bacterial diseases is very difficult due to antibiotic resistance and ineffectiveness of chemical products, to that essential oils offer a promising alternative. Minimal inhibitory and bactericidal concentrations are determined by applying a single drop of oil onto the inner side of each plate cover in macrodilution assays. Among all tested substances, the strongest and broadest activity was shown by the oils of wintergreen (Gaultheria procumbens), oregano (Origanum vulgare), and lemongrass (Cymbopogon flexuosus. Carvacrol (64.0–75.8%) was the dominant component of oregano oils, while geranial (40.7%) and neral (26.7%) were the major constituents of lemongrass oil. Xanthomonas campestris pv. phaseoli was the most sensitive to plant essential oils, being susceptible to 19 oils, while 11 oils were bactericidal to the pathogen. Sixteen oils inhibited the growth of Clavibacter michiganensis subsp. michiganensis and seven oils showed bactericidal effects to the pathogen. The least sensitive species was Pseudomonas tolaasii as five oils inhibited bacterial growth and two oils were bactericidal. Wintergreen, oregano, and lemongrass oils should be formulated as potential biochemical bactericides against different horticultural pathogens.     

不同品种油葵在镉严格管控类耕地的安全利用探索

为探索油葵在镉(Cd)严格管控类耕地安全利用的可行性,采用田间实验,研究了15个品种油葵的生长效应、各部分Cd富集能力差异,并评估了该油葵所产食用油的安全性。结果表明,油葵对Cd有一定的耐性,不同品种油葵地上部生物量在1 605~5 500 kg·hm⁻²;油葵中Cd质量分数在非食用部分的分布为茎叶>花盘>油枯;油枯、花盘和茎叶的生物富集系数(BCF)分别在2.07~5.20、7.03~14.87和4.20~11.56,均表现出较大差异。所有品种油葵食用油部分均未超过欧盟食品限量标准中规定的限值(Cd≤0.1 mg·kg⁻¹),并且其富集能力极低。通过对不同品种油葵Cd富集能力的聚类分析、Cd提取量与修复效率的分析,在Cd严格管控类耕地上边利用、边修复较合适的是Y13和Y4品种。所有品种油葵的茎叶、花盘和油枯的Cd质量分数均超过了国家规定的饲料标准(Cd≤1 mg·kg⁻¹)和有机肥料标准(Cd≤3 mg·kg⁻¹),直接还田存在较高风险,需要另外处理。本研究可为Cd严格管控类耕地的安全利用提供参考。     

金鱼藻对不同扰动方式下悬浮物的生理响应

以金鱼藻为研究对象,分析悬浮物不同扰动方式(机械扰动和曝气扰动)、扰动强度以及悬浮物浓度对沉水植物生理特性的影响。结果表明,与曝气扰动相比,机械扰动对金鱼藻生理特性影响显著,搅拌强度的增大,金鱼藻叶绿素a、叶绿素b、类胡萝卜素含量和POD活性不断下降,60 r/min时下降最快,搅拌强度与POD活性显著负相关。曝气扰动在前期促进金鱼藻的生长,后期抑制其生长,曝气强度4 m3/h时,金鱼藻生理指标降低最大,曝气强度与金鱼藻生理特性相关性不显著。随悬浮物浓度升高,金鱼藻叶绿素a、叶绿素b、类胡萝卜素含量先升高后下降,金鱼藻生长最适悬浮物浓度为0.4~0.8 g/L。     

Toxicity of commercial fungicides in relation to the larvae of Drosophila melanogaster Meigen (Diptera, Drosophilidae) and effect on the sex ratio

Bouillie bordelaise, Benlate, Euparène and Dithane M45 had no effects on Drosophila eggs. However, a post-hatching mortality was observed and, in all cases, a "dose effect" was established. There was a quick effect with Bouillie bordelaise, a slow and continued one with Benlate and Euparène, and the effect was delayed until the pupal stage with Dithane. Bouillie bordelaise, Benlate and Euparène slowed down the larval development and the emergence period was longer, according to the doses. Bouillie bordelaise was always the most toxic fungicide and Dithane the least one. The sex-ratio was not disturbed with Bouillie bordelaise. With Benlate and Euparène, the females were more sensitive than the males, while Dithane was more toxic towards males. Considering a same fungicide, a difference was exhibited according to the chosen criterion: Benlate caused an effect on larval mortality which was more important than its effect on the developing-time. Moreover, both carbamates, Benlate and Dithane, acted differently whatever their toxicity level.     

Toxicity of hydroquinone to different freshwater phototrophs is influenced by time of exposure and pH

The interaction of natural organic matter with phytoplankton communities in freshwater ecosystems is an intensively studied subject matter. Previous studies showed that apparently plant-derived phenols were able to inhibit algal and cyanobacterial growth. Furthermore, it was also assumed that humic substances (HS), which comprise the major part of dissolved organic carbon in freshwater ecosystems, directly interact with freshwater phototrophs. For example, quinoid building blocks of HS were thought to be algicidal. To identify key environmental variable for the toxic action of potential quinone algicides, we tested the toxicity of hydroquinone (HQ) to different eukaryotic and prokaryotic freshwater phototrophs in terms of growth performance and investigated also the effect of HQ oxidation at different pH values on its algicidal potential. It was shown that cyanobacterial species were much more susceptible to hydroquinone than coccal green algal species were, with Microcystis aeruginosa being the most sensitive species by far. In addition, it was obvious that the aging of hydroquinone-stock solution at pH 11 led to polymerization and, by this process, to a total loss of toxicity; whereas the algicidal potential sustained if the polyphenol was kept at pH 7. Since most lakes with heavy blooms of phototrophs possess pH values clearly above 7.0, it is questionable, if polyphenols in general and quinones in particular are the effective chemicals and if litter and straw leachates are applied as means to combat algal and cyanobacterial blooms.     

不同挂膜方式生物滴滤塔处理含H2S恶臭气体简

采用菌剂挂膜,活性污泥挂膜和自然挂膜3种不同方式形成生物滴滤塔,考察挂膜方式对生物滴滤塔去除H₂S恶臭气体的影响。结果表明,当进气H₂S浓度为5 mg/m³时,菌剂挂膜、活性污泥挂膜、自然挂膜形成的生物滴滤塔出气H₂S浓度分别为15.7~17.4、11.6~14.8和15.0~15.9 μg/m³;塔内压降分别为3~4 mm水柱、6 mm水柱和4~5 mm水柱;喷淋后滤出液中硫酸根的浓度分别为14、22和17 mg/L,硫的转化率分别为45%、60%和50%。当进气H₂S浓度增大至7 mg/m³时,3个塔经过7 d的调整后,均能达到稳定状态,稳定后3个塔中出气H₂S浓度和压降基本没变,喷淋后滤出液中硫酸根浓度依次增大至25、31和30 mg/L左右。采用活性污泥挂膜形成的生物滴滤塔处理H₂S的能力比菌剂挂膜和自然挂膜的高。     

不同挂膜方式生物滴滤塔处理含H2S恶臭气体

采用菌剂挂膜,活性污泥挂膜和自然挂膜3种不同方式形成生物滴滤塔,考察挂膜方式对生物滴滤塔去除H2s恶臭气体的影响。结果表明,当进气H2S浓度为5mg／m3时,菌剂挂膜、活性污泥挂膜、自然挂膜形成的生物滴滤塔出气H2s浓度分别为15．7～17．4、11．6～14．8和15．0～15．9μg／m3;塔内压降分别为3—4mm水柱、6mm水柱和4—5mm水柱;喷淋后滤出液中硫酸根的浓度分别为14、22和17mg／L,硫的转化率分别为45％、60％和50％。当进气H2S浓度增大至7mg／m3时,3个塔经过7d的调整后,均能达到稳定状态,稳定后3个塔中出气H2s浓度和压降基本没变,喷淋后滤出液中硫酸根浓度依次增大至25、31和30mg／L左右。采用活性污泥挂膜形成的生物滴滤塔处理H2s的能力比菌剂挂膜和自然挂膜的高。