Action of the fungicide tridemorph on the glucose,lactate and succinate dehydrogenase activities of some tridemorph-sensitive and -resistant bacteria

The systemic morpholine fungicide tridemorph, which is known to exert its antifungal action through inhibition of ergosterol biosynthesis, can also inhibit the growth of organisms which are incapable of sterol biosynthesis. It was found to inhibit strongly glucose and lactate dehydrogenase activities in cultures of four Gram( +) bacteria, Rhodococcus sp. AK 1, Bacillus cereus Frankland & Frankland, Bacillus subtilis (Ehrenberg) Cohn, Nocardia asteroides and a Gram(?) bacterium, Rhizobium leguminosarum. Growth of these bacteria was inhibited by tridemorph at concentrations between 7 and 60 mg litre^?1. In contrast, similar dehydrogenase activities in other Gram(?) organisms, Escherichia coli Cast. & Chalm. and Azotobacter vinelandii, which showed no growth inhibition at 200 mg litre ^?1 tridemorph, were either not inhibited or inhibited only slightly. Similarly, succinate dehydrogenase activity in Rhodococcus sp. AK 1 was more strongly inhibited by tridemorph than that in E. coli. In cell-free extracts of Rhodococcus sp. AK1 and E. coli, lactate dehydrogenase activity was also inhibited by tridemorph to a much greater extent in the sensitive strain (63%) than in the resistant ones (8%).     

嘧菌酯对番茄早疫病菌的抑制作用

采用菌丝生长速率法和孢子萌发法分别测定了嘧菌酯对番茄早疫病菌Alternaria solani菌丝生长的抑制作用及其对分生孢子萌发的影响,同时测定了其对菌丝呼吸速率的影响。结果表明,嘧菌酯抑制番茄早疫病菌分生孢子萌发的作用强于对菌丝生长的抑制作用。抑制菌丝生长的剂量-反应曲线与抑制菌丝呼吸的剂量-反应曲线趋势相似,但菌丝在经嘧菌酯处理5~7 h后呼吸速率恢复到对照水平,相比之下,嘧菌酯却能一直抑制菌丝的生长。因此,采用孢子萌发法测定番茄早疫病菌对嘧菌酯的敏感性比用菌丝生长速率法测定更为适宜。水杨肟酸与嘧菌酯在抑制番茄早疫病菌菌丝生长和分生孢子萌发方面有协同作用,对菌丝生长的平均协同系数为7.24。随处理时间延长,药剂对菌丝耗氧的抑制作用下降,但仍抑制菌丝生长。研究表明:呼吸作用对药剂的敏感性随着处理的延长而下降的机理不是因为旁路氧化途径增强, 也不是因为基质中药剂效力的下降,而是存在其它机制。     

Genetic control of resistance to tridemorph inustilago maydis

Mutants ofUstilago maydis with low resistance to tridemorph isolated in a mutation frequency of 7x 10^-6 after UV-irradiation and selection on media containing 25 μg ml^-1 tridemorph. Genetic analysis with nine such mutant isolates resulted in the identification of two unlinked chromosomal loci,U/tdm- 1 andU/tdm- 2. TheU/tdm mutations are responsible for low resistance levels to tridemorph (resistance factor, Rf, of 3 or 5 based on effective concentration causing a 50% reduction in the growth rate (EC₅₀) or minimal inhibitory concentration (MIC) values, respectively) and low to moderate level of resistance to fenpropimorph (Rf 10 or 16 based on MIC or EC₅₀, respectively) and fenpropidin (Rf 5 or 11 based on MIC or EC₅₀, respectively). Haploid strains carrying bothU/tdm mutations exhibit higher levels of resistance to the above fungicides, indicating interallelic interaction between nonallelic genes. Crosses between mutants carrying theU/tdm- genes with compatible isolates carrying theU/fpm- 1 orU/fpm- 2 mutations, which were found in previous work to carry fenpropimorph resistance, yielded in all cases a large number of recombinants with wild-type sensitivity, indicating that the mutant genes involved were not allelic. Cross-resistance studies with the inhibitors of C-14 demethylase showed that. the U/tdm-mutations were responsible for increased sensitivity to the triazoles triadimefon, triadimenol, propiconazole and flusilazole, and to the pyridine pyrifenox. Study of gene effect on the fitness ofU. maydis showed thatU/tdm-mutations appeared to be pleiotropic, having more or less adverse effects on growth rate in liquid culture and pathogenicity on young corn plants.     

Antifungal activity of metalaxyl and furalaxyl

The systemic acylalanines metalaxyl and furalaxyl proved in vitro highly selective to fungi of the order of the Peronosporales, the former being somewhat more active than the latter. Various other fungal, and bacterial, species did not show sensitivity to the toxicants. Metalaxyl was strikingly more active on Pythium splendens and Phytophthora cinnamomi at low than at high temperature. With Phytophthora species metalaxyl displayed improved activity in combination with sodium dodecylsulphate and Triton X-100. Neither the composition of the medium nor its pH influenced activity. None of many compounds tested could antagonize their effect. The fungitoxicity caused by metalaxyl in the gas phase was strikingly better than that of furalaxyl against P. splendens. The toxicants had little effect on germination of the hyphal swellings of P. splendens, but had significant influence on the subsequent hyphal development. This included reduced growth, abnormal variation in the fungal thickness, and inhibition of the formation of hyphal swellings. Metalaxyl and furalaxyl were fungistatic at minimal inhibitory concentration (MIC) values during the first 24 hr. Thereafter, the toxicants became fungicidal. Metalaxyl and furalaxyl did not inhibit respiration of P. splendens at MIC values or somewhat higher concentrations. Neither did metalaxyl inhibit the [¹⁴C]carbon dioxide evolution from externally supplied radiolabeled glucose.     

Studies on the effects of herbicides on soil nitrification,II

The effects of selected herbicides have been studied on the following parameters of soil nitrification processes: the rates of nitrate and nitrite formation from ammonia in freshly perfused soils and in soils previously saturated with nitrifying organisms in an improved perfusion apparatus; the rates of oxygen consumption and of oxidation of ammonia and nitrite in washed cell suspensions of Nitrosomonas europaea and Nitrobacter winogradskii, respectively; the rates of growth of those two organisms in newly established cultures; the rates of oxygen uptake by soil enriched in nitrifying organisms; the rates of proliferation of nitrifying populations in freshly perfused soil. ID50 values were computed for all parameters and herbicides studied. On average, the most sensitive parameters were the metabolism and growth of the two organisms in pure culture, while the least sensitive were the corresponding measures in the soil environment. Similarly, herbicides fell into four distinct groups. The most toxic were the formulated octanoates of bromoxynil and ioxynil (NPH1320 and Totril, respectively); next in order of toxicity were chlorbufam, phenmedipham, formulated oxadiazon, formulated legurame, ioxynil, formulated trifluralin, and bromoxynil; low toxicity was shown by terbacil, dicamba, and tricamba, whereas asulam and the related experimental herbicide MB9555 showed activity on some parameters at the very highest concentrations only. Comparisons of soil with pure-culture parameters showed that the relative toxicities of herbicides to Nitrosomonas in culture bore little relationship to those in soil. The inhibitions of Nitrobacter proliferations in soil on the other hand were correlated with the inhibitions of growth and metabolism in pure culture. Within these overall effects, individual herbicides showed marked differential actions on various parameters. Thus, the formulated octanoates of bromoxynil and ioxynil were extremely toxic to the growth of nitrifying organisms in culture, an action probably due to an unknown formulation component. Relative to other herbicides, bromoxynil and ioxynil were more effective on nitrification processes in the soil environment. Both in culture and in the soil, Nitrobacter is more sensitive than Nitrosomonas to these four herbicides. Legurame and oxadiazon are relatively more toxic to Nitrosomonas in culture, but this differential action is not demonstrable in the soil. Dicamba, tricamba, trifluralin, and chlorbufam are more toxic to Nitrobacter than to Nitrosomonas in the soil environment. Formulated trifluralin seems to exert a stimulating action on the growth of nitrifying organisms, but only in the soil; suppression of antagonistic organisms is suggested as a possible cause. Extrapolation of these results to the field situation suggests that the only herbicide which might cause small inhibitions of nitrification at field rates is terbacil, which is disproportionately toxic at low concentrations. At rates somewhat in excess of normal, the formulated octanoates of bromoxynil and ioxynil and possibly dicamba and oxadiazon could also cause small inhibitions.     

Trifluralin effects on carrot callus tissue

The effect of trifluralin on the growth, oxygen uptake, and adenosine phosphates level of carrot (Daucus carota L.) callus tissue was determined over a period of 18 days after subculture. The herbicide (10^?4 and 10^?5M) reduced fresh weight gain significantly; the reduction was less with lower trifluralin concentrations. Dry weight accumulation was not inhibited until after Day 6 of the test period and thereafter was reduced by all concentrations tested. Oxygen consumption was inhibited by trifluralin (10^?4 and 10^?5M) throughout the test period. Concentrations of 10^?6.10^?7, and 10^?8M produced variable effects. The response profile of O₂ consumption in the presence of dinitrophenol was different from that of trifluralin. Analysis of adenosine phosphates level gave no clear response trend. Energy charge values of 0.7 to 0.85 were obtained for untreated tissue. Trifluralin had no effect on energy charge until Day 9 (10^?4M) and after Day 15 (all concentrations).     

Uptake by roots and translocation to shoots of two morpholine fungicides in barley

Despite being lipophilic, morpholine fungicides are systemic in plants. Such transport may be explicable by their protonation (pKa∽7·5) at the pH of plant compartments to yield the more polar cation. This behaviour might be a useful attribute to be incorporated into other classes of lipophilic pesticides. To understand quantitatively the behaviour of the morpholine fungicides, the uptake by roots and transport to shoots in barley of two such ¹⁴C-labelled compounds, dodemorph and tridemorph, were investigated using bathing solutions of differing pH. At pH 5, uptake and transport were small, but increased by approximately two orders of magnitude at pH 8. Tridemorph, the more lipophilic of the two compounds, was highly accumulated by roots at pH 8 and moderately translocated to shoots. In contrast, dodemorph was translocated to shoots at pH 8 with remarkable efficiency, moving into the xylem across the endodermis at 23 times the efficiency of water, though accumulation in roots was less than that of tridemorph. Behaviour at 24 h was largely similar to that at 48 h for both compounds, indicating that uptake and translocation are equilibrium processes maintained over time. Transport to shoots for each compound was directly proportional to the concentrations accumulated in the roots, except at low pH where partitioning into root solids became proportionately more important with such material not being directly available for transport to the xylem across the endodermis. Uptake and transport of these basic fungicides are explained in terms of their partitioning and of their accumulation in acidic plant compartments by ion trapping as the protonated form; this behaviour is discussed in relation to the pKa and lipophilicity of these compounds. © 1998 Society of Chemical Industry     

The fungicide tridemorph as a selective herbicide for the control of Holcus lanatus in ryegrass and of Bonus sterilis in barly

The potential of the fungicide tridemorph selectively to control established plants of Holcus lanatus L. in ryegrass (Lolium perenne L) and Bromus sterilis L. in barley (Hordeum vulgare L.) was examined in glasshouse trails. A dose of 4.2 kg ha^?1 tridemorph gave selectively between H. lanatus and ryegrass similar to, but more costly than, that which would be provided by 1.2 kg ha^?1 asulam. B sterilis was more sensitive to tridemorph than was barley. Two additives, glycerol and the oil-surfactant mixture ‘PF’ enhanced this selectivity when tridemorph was applied at 2.1 kg ha^?1.     

Inhibitory effect of bionic fungicide 2‐allylphenol on Botrytis cinerea (Pers. ex Fr.) in vitro

BACKGROUND: 2‐Allylphenol is a registered fungicide in China to control fungal diseases on tomato, strawberry and apple. It is synthetic and structurally resembles the active ingredient ginkgol isolated from Ginkgo biloba L. bark. 2‐Allylphenol has been used in China for 10 years. However, its biochemical mode of action remains unclear. An in vitro study was conducted on the biochemical mechanism of 2‐allyphenol inhibiting Botrytis cinerea (Pers. ex Fr.). RESULTS: The inhibition was approximately 3 times stronger when the fungus was grown on non‐fermentable source, glycerol, than that on a fermentable carbon source, glucose. Inhibition of B. cinerea and Magnaporthe oryzae (Hebert) Barr mycelial growth was markedly potentiated in the presence of salicylhydroxamic acid (SHAM), an inhibitor of mitochondrial alternative oxidase. Furthermore, at 3 h after treatment with 2‐allylphenol, oxygen consumption had recovered, but respiration was resistant to potassium cyanide and sensitive to SHAM, indicating that 2‐allylphenol had the ability to induce cyanide‐resistant respiration. The mycelium inhibited in the presence of 2‐allylphenol grew vigorously after being transferred to a fungicide‐free medium, indicating that 2‐allylphenol is a fungistatic compound. Adenine nucleotide assay showed that 2‐allylphenol depleted ATP content and decreased the energy charge values, which confirmed that 2‐allylphenol is involved in the impairment of the ATP energy generation system. CONCLUSION: These results suggested that 2‐allylphenol induces cyanide‐resistant respiration and causes ATP decrease, and inhibits respiration by an unidentified mechanism. Copyright © 2009 Society of Chemical Industry     

Effect of Germination Initiation on Competitive Capacity of Trichoderma atroviride P1 Conidia

ABSTRACT Trichoderma biocontrol isolates are most effective as highly concentrated inocula. Their antagonism to other fungi may be a result of pregermination respiration. In a nutrient-rich medium, almost all Trichoderma atroviride P1 (P1) conidia initiated germination processes and increased respiration, even in dense suspensions. When 1 x 10(7) P1 conidia/ml were coinoculated with 1 x 10(5) Botrytis cinerea conidia/ml, dissolved oxygen fell to <1% within 2 h and the pathogen failed to germinate. More dilute P1 suspensions consumed oxygen slowly enough to allow coinoculated B. cinerea to germinate. On nutrient-poor media, fewer P1 conidia initiated germination. Oxygen consumption by the inoculum and inhibition of B. cinerea were enhanced when P1 conidia were nutrient activated before inoculation. Pregermination respiration also affected competitive capacity of the antagonist on solid substrates, where respiratory CO(2) stimulated germination rate and initial colony growth. These parameters were directly correlated with inoculum concentration (R(2) >/= 0.97, P < 0.01). After initiating germination, Trichoderma conidia became more sensitive to desiccation and were killed by drying after only 2 h of incubation on a nutrient-rich substrate at 23 degrees C. These results indicate that nutrient-induced changes preceding germination in Trichoderma conidia can either enhance or decrease their biological control potential, depending on environmental conditions in the microhabitat.     

Resistance of Erysiphe graminis on barley and wheat to sterol C-14-demethylation inhibitors1

Isolates of Erysiphe graminis f. sp. hordei and tritici with decreased sensitivity to triadimefon showed cross-resistance to other inhibitors of sterol C-14-demethylation, such as triadimenol, propiconazol, diclobutrazol, prochloraz and nuarimol. The isolates exhibited a moderate degree of resistance to these compounds. No cross-resistance was detected to tridemorph, fenpropimorph and pyrazophos. The resistant hordei isolates were more sensitive to ethirimol than the sensitive isolate. The competitive abilities of resistant hordei and tritici isolates were inferior to that of the sensitive isolates. In the presence of the fungicides no differences in germination, appressorium formation and penetration between the sensitive and resistant isolates were observed; 48 h after inoculation the sensitive isolate showed several morphological alterations and further fungal development was arrested. At four to five times higher doses of triadimefon, similar morphological alterations were detected in the resistant isolate. Low concentrations of triazole fungicides which slightly affected mycelium growth of both the sensitive and the resistant isolate of f.sp. hordei severely inhibited development of conidiophores of the sensitive isolate whereas that of the resistant isolate was hardly affected.     

On the antifungal mode of action of tridemorph

Tridemorph (2,6-dimethyl-N-tridecylmorpholine) was active against representative of nearly all taxonomic groups of fungi; gram-positive bacteria were also sensitive although gram-negative were not. Tridemorph, 3–10 μg/ml, inhibited the multiplication of sporidia of Ustilago maydis more strongly than the increase of dry weight. The treated sporidia appeared swollen, multicellular, and sometimes branched. Unsaturated lipophilic compounds like α-tocopherol and trilinolein alleviated the toxicity of tridemorph to Botrytis allii and U. maydis. Protein and RNA syntheses were inhibited slightly. DNA synthesis was rather strongly affected already after 2 hr. Lipid synthesis was first inhibited but later stimulated. At an early stage (2 hr) treated cells differed already from control cells by a higher content of free fatty acids. Tridemorph also inhibited sterol biosynthesis. The antimicrobial spectrum, the characteristic morphology of treated cells of U. maydis, the observations on cross-resistance, the alleviating effect of unsaturated lipophilic compounds, and the alterations in neutral lipid pattern suggest strong similarity of the mode of action of tridemorph with that of the known inhibitors of sterol biosynthesis.     

Growth of facultative anaerobic population of human intestinal bacteria with the carbamate pesticide aminocarb

The effect of different concentrations of the carbamate pesticide, aminocarb (Matacil), on the growth and selection of facultative anaerobic bacteria and degradation of the pesticide by human endoflora of the intestinal tract was examined in vitro. Microorganisms were cultured under aerobic or anaerobic conditions, in nutrient broth and mineral media. The intestinal population was more sensitive to 10–1000 μg/ml aminocarb under anaerobic conditions than in aerobic culture; however, spontaneous degradation of aminocarb in media markedly affected the degree of bacterial growth inhibition in prolonged cultures. In addition, the type of culture medium appeared to influence the degree of aminocarb-induced bacterial growth inhibition. A dose of aminocarb inducing 50% growth inhibition was established for different culture conditions: for mineral medium, aminocarb inhibited bacterial growth by 50% at 600 μg/ml under anaerobic and aerobic conditions, whereas less than 50% inhibition was observed even at 1000 μg/ml aminocarb when bacteria were grown in nutrient broth. A selection of bacterial strains occurred in the presence of increasing aminocarb concentrations, which was determined quantitatively and qualitatively by the identification of codominants. A shift in several Escherichia coli biotypes was also observed in cultures with aminocarb, in comparison to control cultures. Bacterial degradation of aminocarb, under anaerobic and aerobic conditions, was determined in a mixed population of the intestinal microflora by high-performance liquid chromatography analysis of culture media. Data showed that aminocarb can be quickly degraded by human intestinal bacteria at relatively high pesticide concentrations. Moreover, other HPLC data suggest rapid spontaneous degradation of aminocarb in neutral and slightly alkaline pH conditions characteristic of the human intestinal tract, which can effectively eliminate the pesticide. Therefore, aminocarb, at the concentrations used, does not seriously affect the bacterial microflora of the human gut.     

Antifungal activity and release of compounds on Rhizopus stolonifer (Ehrenb.:Fr.) Vuill. by effect of chitosan with different molecular weights

The effect of chitosan on mycelial growth, protein release, glucose consumption, hexokinase activity and the release of compounds was studied in three isolates of Rhizopus stolonifer. The chitosan of low, medium and high molecular weight at all evaluated concentrations inhibited the mycelial growth in the three isolates of R. stolonifer. In presence of any type of chitosan the release of proteins was notably increased. Medium molecular weight chitosan showed a major effect on the release of proteins when compared with the other two types of chitosan. The glucose consumption in presence of chitosan with different molecular weight was increased significantly in the three isolates of R. stolonifer. The highest glucose consumption was induced with chitosan of low molecular weight. Hexokinase activity was stimulated at least 2-fold in presence of any types of chitosan. The three isolates showed an increased release of compounds at 260 and 280 nm with chitosan of different molecular weight. These results suggest that all types of chitosan showed antifungal activity and increased significantly the glucose consumption and the release of compounds in the three isolates of R. stolonifer.     

Benomyl and methyl-2-benzimidazolecarbamate (MBC): Biochemical,cytological and chemical aspects of toxicity to Ustilago maydis and Saccharomyces cerevesiae

Action of methyl-2-benzimidazolecarbamate (MBC), a breakdown product of benomyl, was studied in synchronous cultures of Ustilago maydis and Saccharomyces cerevesiae. Cells treated with MBC developed similarly to control cells until the two portions of the doublets (joined mother and daughter “cells”) were morphologically equivalent. At this point development of the treated cells ceased but control doublets separated to form two new cells. The compound did not prevent DNA synthesis during the first cell cycle of synchronized S. cerevesiae cells but did prevent completion of cell division, as it did also in synchronized U. maydis sporidia which already contained the DNA complement needed for completion of the first division. Mitosis did not go to completion in MBC-treated cells so the doublets formed contained only a single compact nucleus. The effect of MBC markedly resembles mitotic arrest caused by colchicine and isopropyl N-phenylcarbamate in higher plants and griseofulvin in the fungus Aspergillus nidulans. Inhibition of DNA synthesis in U. maydis and inhibition of cytokinesis in both organisms studied are secondary effects attributable to mitotic failure.A volatile compound toxic to U. maydis and S. cerevesiae was demonstrated in air over moistened benomyl. Hexane, through which such air samples were passed, or hexane extracts of aqueous suspensions of benomyl contained a substance having the same ir spectrum and retention time in two gas chromatographic systems as butyl isocyanate (BIC).BIC inhibited respiration of U. maydis and S. cerevesiae in a manner similar to benomyl. Benomyl partially inhibited respiration of subcellular particles from both organisms. MBC had essentially no effect on whole cell or subcellular particle respiration of either organism. Toxicity of benomyl preparations is not attributed to “benomyl” as such, but to two breakdown products, MBC and BIC. Therefore, differential effects of benomyl preparations and of MBC on growth and metabolism in fungi should be ascribed to BIC.     

Bleaching activity and chemical constitution of phenylpyridazinones

The bleaching effect of 2-phenylpyridazinones substituted at the 4 or 5 position of the pyridazinone moiety or at the phenyl ring (position 9) was assayed using a greening Scenedesmus mutant after its transfer from heterotrophic to autotrophic growth. The following relationship between bleaching activities and Hammett electronic parameters of the various substituents could be demonstrated. The biological activity of the pyridazinone skeleton was enhanced with substituents showing (a) increasing σ_m values in position 4, (b) increasing σ_m or σ_p values in position 9, and (c) decreasing σ_p values in position 5. These findings could be corroborated by data on pigment bleaching and decrease of photosynthetic oxygen evolution of autotrophic (green) wild-type Scenedesmus after growth in the presence of sublethal concentrations of pyridazinones. There is no structure/activity relationship with direct inhibition of photosynthetic electron transport. Based on electronic parameters, the construction of phenylpyridazinone derivatives with bleaching activity is proposed.     

Mitochondrial electron transport in a carboxin-resistant,antimycin A-sensitive mutant of Ustilago maydis

Succinoxidase activity of mitochondria from an antimycin A-sensitive (ants) mutant of Ustilago maydis is approximately five times less sensitive to the fungicide carboxin than that of mitochondria from wild-type cells. The antimycin A sensitivity is due to the absence of an alternative electron transport pathway in mitochondria obtained from mutant cells grown in control medium. The same mutant, however, develops high rates of alternative respiration if grown in the presence of chloramphenicol. Substrate and oxygen affinity as well as resistance to hydroxamates indicate that this respiration is mediated by only one mitochondrial electron transport pathway, similar to the inducible system described earlier in wild-type mitochondria. Induction appears to be regulated by the activity of the cytochrome pathway. The absence of the constitutive system from the ants mutant mitochondria which are resistant to carboxin, a selective inhibitor of the succinate dehydrogenase complex, supports the view that this system must be related to succinic dehydrogenase.     

Effects of herbicidal carbamates on mitochondria and chloroplasts

At concentrations near 2 × 10^?4M, barban, chlorpropham, and phenmedipham are inhibitors of the electron transfer in potato and mung bean mitochondria. The inhibition seems to be localized in the flavoprotein region. It affects preferentially the exogenous NADH dehydrogenation, in potato mitochondria (I₅₀, 10^?4M). Succinate dehydrogenation is less inhibited. At noninhibiting concentrations, the studied carbamates cannot uncouple the oxidative phosphorylations. Photosynthesis is completely inhibited by 2.10^?7M phenmedipham, 5 × 10^?5M barban, and 2 × 10^?4M chlorpropham. The inhibition takes place at the PS II level. Moreover, barban and chlorpropham are uncouplers of the photophosphorylations for concentrations between 5 × 10^?5 and 5 × 10^?4M. The effects observed on mitochondrial respiration can also be found on respiration of Acer cultured cells. The effects on isolated chloroplast photosynthesis are also observed for slightly higher concentrations on cultured Chlorella and on pea and oat leaf fragments.     

Effects of the organophosphate pesticide Folidol 600® on the freshwater fish, Nile Tilapia (Oreochromis niloticus)

Nile Tilapia (Oreochromis niloticus) juveniles were exposed to different concentrations of Folidol 600® in static toxicity tests. The 24, 48, 72 and 96 h LC₅₀ values of Folidol 600® to O. niloticus were 17.82, 8.91, 4.00 and 2.70 mg L⁻¹, respectively. The values of hematological parameters increased, and inhibition of cholinesterases activity (AChE, BChE and PChE) in plasma of fish exposed to the higher concentrations of pesticide reached 94%. Furthermore, the exposure of Tilapia to Folidol 600® caused an increase of 4%, 20% and 38.4% in oxygen consumption at 0.1, 0.5 and 1.0 mg L⁻¹, respectively. However, exposure to 2.5, 5.0 and 10 mg L⁻¹ caused a decrease of 33.6%, 35.2% and 42.4% in oxygen consumption relative to the control. The ammonium excretion of fish exposed to 0.0, 0.1, 0.5, 1.0, 2.5, 5.0 and 10.0 mg Folidol 600®/L was 0.12, 0.18, 0.30, 0.33, 0.37, 0.36 and 0.33 μg/g/min, i.e., 50%, 150%, 175%, 208%, 200% and 175% increase, respectively, relative to the control.     

Specific effects of tridemorph on sterol biosynthesis in Ustilago maydis

In an attempt to indicate the site of action of tridemorph in ergosterol biosynthesis by Ustilago maydis the nature of the sterol intermediates accumulating in treated cells was studied. At low growth-inhibiting concentrations of the toxicant (10 and 15 μg/ml) decline of ergosterol content during 6 hr of incubation was accompanied by an accumulation of various sterol intermediates of which ergosta-8,14-dien-3β-ol appeared to be the major sterol. After isolation of this intermediate its identity was further confirmed by direct comparisonof its ultraviolet, glc, and mass spectrum with those of an authentic synthesized sample of ergosta-8,14-dien-3β-ol (ignosterol). Results indicate that toxicity of tridemorph is caused by a specific inhibitive effect on the enzyme Δ¹⁴-reductase which is responsible for $\text{14}\text{15}$ double-bond reduction in sterol biosynthesis.