Effects of fungicide residues on the survival, fecundity, and predation of the mites Tetranychus urticae (Acari: Tetranychidae) and Galendromus occidentalis (Acari: Phytoseiidae)

Representative fungicides from three or four families used for management of powdery mildew and other diseases in tree fruits were evaluated for their effects on a common spider mite and predator mite species, respectively. A modified Munger cell technique was effective in measuring the response of phytophagous and predaceous mites to fungicide residues on detached leaves in the laboratory. Demethylation-inhibiting (DMI) (imidazole [triflumazole] and triazole [myclobutanil]) and strobilurin (trifloxystrobin) fungicides were not toxic to female Tetranychus urticae Koch and Galendromus occidentalis (Nesbitt), and no sublethal effects were found on fecundity and predation rate after 3-5-d exposure to residues. Benomyl, a benzimidazole fungicide, increased adult mortality and reduced fecundity for both mite species; however, it did not alter the predation rate of G. occidentalis females on T. urticae eggs and larvae. Female G. occidentalis that survived the lethal effects of benomyl and the comparison acaricide pyridaben were unimpaired in predation. Our results for benomyl substantiate those of earlier studies and provide evidence for nontoxic effects of DMI and strobilurin fungicides on mites. We propose that DMI and strobilurin fungicides are a good fit for integrated mite management programs due to conservation of phytoseiid predatory mites.     

Influence of fungicides on grape yeast content and its evolution in the fermentation

The influence of six fungicides (famoxadone, fenhexamid, fluquinconazole, kresoxim-methyl, quinoxyfen, trifloxystrobin) on the yeast content in harvested grapes has been studied along with their effect on it during the wine-making process. Two treatments were carried out with authorized formulates at the manufacturer doses. The first was carried out under good agricultural practices, obeying the security times, and the second one under critical conditions, applied on the day of harvesting. The grapes were harvested two hours after the application. During the wine-making process, samples were taken at 1, 5, 12 and 20 days after the start. The levels observed in the control sample (1.9 log CFU/cm2) were similar to previous studies. The counts in all treated samples were higher than the control sample, without adverse effect of the pesticides on the yeast count even in the grapes treated on the day of harvest. As with the observations on grapes, no effect of the pesticides on the fermentation count was observed. All counts on the treated samples were higher than the control sample. The treatment on the day of harvest did not have any effect on the fermentation count, with superior results being obtained for all the pesticides.     

Effects of fluquinconazole and silthiofam, applied as seed treatments to single or consecutive crops of wheat, on take-all epidemic development and grain yields

Seed treatments containing fluquinconazole, silthiofam or a standard fungicide mixture with no activity against take‐all were compared in all combinations of sequences in successive second and third winter wheat crops in five field experiments and second to fourth crops in a sixth experiment. Compared with the standard treatment, silthiofam decreased take‐all more effectively than fluquinconazole when crops were sampled at tillering. In samples taken in summer, during grain filling, silthiofam often decreased the incidence of take‐all (percentage of plants with root symptoms) more than fluquinconazole, but fluquinconazole more effectively decreased the incidence of severe take‐all (percentage of plants with more than 75% of their root systems blackened). It is suggested that these differences are a consequence of more effective control of primary infection of roots by silthiofam and of secondary, root‐to‐root, infection by fluquinconazole. Silthiofam usually increased yield more than did fluquinconazole, perhaps as a consequence of better early protection during tiller and/or spikelet formation. Treatment with either of the fungicides affected epidemic development in the treated crop and in crops grown subsequently. In particular, decreased take‐all had the effect of delaying the year‐to‐year epidemic, so that nontreatment of a subsequent crop resulted in an upsurge in disease. Treatment with either take‐all fungicide of a crop grown after a treated crop was relatively effective if the epidemic in the comparable nontreated crop sequence was continuing to increase. It was, however, detrimental if the disease was approaching its peak in the first treated crop, particularly if a treated (fourth wheat) crop was being compared with a similar crop in a nontreated sequence in which take‐all decline had developed. These results provide a basis for recommendations for the use of seed treatment fungicides in sequences of wheat crops.     

Impact of winemaking practices on arginine and citrulline metabolism during and after malolactic fermentation

AIMS: To study arginine degradation and carcinogenic ethyl carbamate precursor citrulline formation during and after malolactic fermentation (MLF). METHODS AND RESULTS: MLF was induced in white wine with two commercial Oenococcus oeni strains under different winemaking conditions regarding the type of alcoholic fermentation (spontaneous, induced) and the lees management (racked, on lees). Arginine degradation and citrulline formation did not occur during malic acid degradation in any treatment. In five of the six treatments in which arginine degradation took place, it occurred 3 weeks after malic acid depletion and significant amounts of citrulline were formed. Presence of yeast lees in wines led to increased citrulline formation. Conclusions: This study suggests that arginine metabolism is inhibited in oenococci at low pH values (< 3.5) and that in the postalcoholic fermentation phase, citrulline formation from arginine degradation can be avoided if MLF is induced by pure cultures of O. oeni with inhibition of the bacterial biomass after malic acid depletion. Residual yeast lees in the wine have been identified as a significant risk factor for increased citrulline formation. SIGNIFICANCE AND IMPACT OF THE STUDY: Conclusions drawn from this study allow reducing the risk of carcinogenic ethyl carbamate formation from citrulline excretion by wine lactic acid bacteria.     

Activity of Ten Fungicides against Phytophthora capsici Isolates Resistant to Metalaxyl

Pepper Phytophthora blight (PPB), caused by Phytophthora capsici, is an important disease of pepper in China. The extensive application of metalaxyl has resulted in widespread resistance to this fungicide in field. This study has evaluated the activities of several fungicides against the mycelial growth and sporangium germination of metalaxyl‐sensitive and metalaxyl‐resistant P. capsici isolates by determination of EC₅₀ values. The results showed that the novel carboxylic acid amide (CAA) fungicide mandipropamid exhibited excellent inhibitory activity against PPB both in vitro and in vivo, with averagely EC₅₀ values of 0.075 and 0.004 μg/ml in mycelial growth and sporangium germination, respectively, and over 88% efficacy in controlling PPB. The other three CAA fungicides also provided over 70% efficacy in controlling PPB. The mycelial growth was less sensitive to quinone outside inhibitor (QoI) fungicides azoxystrobin and trifloxystrobin than that of sporangium germination in P. capsici isolates. However, azoxystrobin and trifloxystrobin provided over 80% efficacy in controlling PPB. It was noted that propamocarb and cymoxanil did not exhibit activity against the mycelial growth or sporangium germination of P. capsici isolates in the in vitro tests, with over 70% efficacy in controlling PPB. The new fungicide mixture 62.5 g/l fluopicolide + 625 g/l propamocarb (trade name infinito, 687.5 g/l suspension concentrate (SC)) produced over 88% efficacy in controlling PPB caused by both metalaxyl‐sensitive and metalaxyl‐resistant isolates. The data of this study also proved that there was obviously no cross‐resistance between metalaxyl and the other tested fungicides. Therefore, these fungicides should be good alternatives to metalaxyl for the control of PPB and management of metalaxyl resistance.     

Effect of a Short Contact Time With Lees on Volatile Composition of Airen and Macabeo Wines

Volatile compounds were analyzed in Airen and Macabeo wines at the end of the alcoholic fermentation and after a short time contact with wine lees. The concentration of 34 analyzed compounds, with the exception of hexyl acetate, linalool, ß-ionone and farnesol, increased significantly after contact with lees in Airen wines. Esters and terpenic compounds decreased significantly in Macabeo wines after contact with lees. The contact with lees could be considered as favourable in Airen wines to improve the aroma quality.     

Influence of pesticides on yeasts colonizing leaves

The effect of nine different pesticides on the growth of yeasts isolated from the leaves of fruit and forest trees was investigated. Four insecticides (with the active ingredients: thiacloprid, deltamethrin, lambdacyhalothrin, and thiamethoxam) and five fungicides (with the effective substances: bitertanol, kresoxim-methyl, mancozeb, trifloxystrobin, and cupric oxychloride) were tested. The concentrations of chemicals were those recommended by the manufacturers for the spraying of trees. The yeast strains isolated from the leaves of fruit trees were not sensitive to any of the insecticides. The majority of yeast strains isolated from the leaves of forest trees were either not sensitive or only to a small extent. While Rhodotorula mucilaginosa and Pichia anomala were not affected by any insecticide, the strains of Cryptococcus laurentii and Rhodotorula glutinis showed the highest sensitivity. The effects of fungicides on the growth of isolated yeasts were more substantial. The fungicide Dithane DG (mancozeb) completely inhibited the growth of all yeasts. All strains isolated from fruit tree leaves were more resistant to the tested fungicides than those isolated from the leaves of forest trees. The most resistant strains from the leaves of fruit trees belonged to the species Metschnikowia pulcherrima, Pichia anomala, and Saccharomyces cerevisiae, whereas Cryptococcus albidus and C. laurentii, originating from the leaves of forest trees, showed the highest sensitivity to fungicides.     

Effects of pesticides on yeasts isolated from agricultural soil

The effect of six various pesticides on the growth of yeasts isolated from agricultural soil was investigated. Two herbicides (with the effective substances lactofen and metazachlor), two fungicides (with the effective substances fluquinconazole and prochloraz), and two insecticides (with the effective substances cypermethrin + chlorpyrifos and triazamate) were tested. It is evident that there are considerable differences in inhibition effects of studied pesticides. The fungicide with the effective substance prochloraz inhibited the growth of majority of yeast strains. The insecticide triazamate at concentration 0.6 mM restricted or inhibited growth of all tested strains. The strains of the genus Cryptococcus were the most sensitive to pesticides, while the strains of the species Cystofilobasidium capitatum, Debaryomyces occidentalis var. occidentalis, and Trichosporon cutaneum were the most resistant.     

四种杀菌剂对玉米螟赤眼蜂酚氧化酶活性的影响

为明确杀菌剂对玉米螟赤眼蜂Trichogramma ostriniae Pang et Chen毒性大小及其成蜂体内酚氧化酶（phenoloxidase, PO）活性影响, 本实验通过测定三唑酮（triadimefon）、 肟菌脂（trifloxystrobin）、 咪鲜胺（prochloraz）、 申嗪霉素（phenazino-1-carboxylic acid）4种杀菌剂对玉米螟赤眼蜂的急性毒性, 并分别测定活体和离体条件下各杀菌剂处理后酚氧化酶的比活性。结果表明, 各药剂对赤眼蜂的急性毒性大小为： 三唑酮＞咪鲜胺＞肟菌酯＞申嗪霉素, 其LC₅₀值分别为3.27, 6.46, 4.06和 9.72 mg/L, 其安全系数依次为0.07, 0.10, 0.20和1.47, 其中申嗪霉素为中等风险性, 其余3种药剂为高风险性, 且杀菌剂明显激活了PO的活性。活体条件下亚致死剂量处理得到的PO比活力高于离体处理, 且三唑酮处理组具有明显的剂量效应。本实验为研究杀菌剂对赤眼蜂的毒性及对PO的影响作了初步的探索, 为进一步研究赤眼蜂对杀菌剂的抗性及免疫能力奠定基础。     

The crystal structure of mitochondrial cytochrome bc1 in complex with famoxadone: the role of aromatic-aromatic interaction in inhibition

Ubiquinol cytochrome c oxido-reductase (EC. 1.10.2.2, bc1) is an integral membrane protein complex essential to cellular respiration. Structures of the 11-subunit mitochondrial bc1 complex were determined with and without the fungicide famoxadone. Specific inhibition by famoxadone is achieved through a coordinated optimization of aromatic-aromatic interactions where conformational rearrangements in famoxadone and in residues lining the inhibitor-binding pocket produce a network of aromatic-aromatic interactions that mimic the crystal lattice of benzene. The profound aromatic-aromatic interactions as supported by prior mutagenesis provide a structural basis for specific protein-ligand interaction in a hydrophobic environment. Dramatic conformational changes, both in cyt. b and ISP subunits in the inhibitor-protein complex, confer experimental evidence for a functional role of cytochrome b in the induced conformational arrest of ISP and allow the identification of a possible intrasubunit signal transduction pathway that controls the movement of ISP. These results support an inhibitory mechanism that is consistent with the requirement for ISP movement in the electron transfer of this complex.     

Effect of numbers of fungicidal sprays and dates of application on control of celery leaf spot (Septoria apiicola) and on chemical residues

Experiments at two sites in each of three seasons showed that successful control of celery leaf spot depended more on the choice of fungicide than on the number of sprays and the dates of application. Organotin fungicides gave best control of celery leaf spot and with one exception residues were not detected even when the last application was given 4 weeks before harvest rather than 6 weeks.     

Chemiluminescent determination of total antioxidant capacity during winemaking.

The total antioxidant capacity (TAC) of five different wines (four red and one white) was determined in five different steps of winemaking carried out in a commercial wine cellar by a chemiluminescence (CL) assay. The CL method is suitable to determine the antioxidant capacity of beverages, and preliminary trials showed that the TAC immediately after the bottle was opened was greater than the day after (about 25% decrease). Immediate analysis or a correct sample storage is therefore necessary. The wines were characterized by different levels of total phenolics and TAC: these differences were related to grape composition and winemaking technologies. The TAC values were the highest immediately after devatting. The TAC suffered the highest decrease (30-50%) after the clarification procedure, which may be due to the fining agents used and to oxygen contact, then remained more or less constant in the subsequent steps.     

The biological evaluation of Zato 50 WG in protection of carrot against Alternaria blight

Effectiveness of new fungicide Zato 50 WG (biologically active substances BAS - trifloxystrobin 50%) in the control of Alternaria blight on carrot was studied. Field experiments were carried out in the years 2003-2004 on carrot cv. Koral. As the standard fungicide Amistar 250 SC (BAS - azoxystrobin 250 g/dm3) was used. The results of health status of carrot leaves analysis in first and second year of experiment showed, that development of disease symptoms was lower on leaves of chemical protected plants than on leaves of control plants. The last estimation of infestation degree of carrot leaves made immediately before harvest showed, that standard fungicide Amistar had longer systemic influence on plants. This was confirmed the lower (statistically significant) infestation index of protected carrot in combination with Amistar. The estimations of health status of carrot roots were made directly after digging up indicate, that there wasn't difference of value of infestation index between protected combination and control. Experiment showed, significant effect of tested fungicides on crop quantity of carrot roots per 1 m2. Especially in the second year of study the crop of carrot roots in combination with Zato was higher - statistically significant - than in the rest combinations.     

Splash dispersal of fungus spores and fungicides in the laboratory and greenhouse

A laboratory technique is described for the production of drops of simulated rain in which fungal spores were suspended. When such drops containing conidia of Botrytis fabae impacted on a target leaf the secondary droplets produced infections on receptor broad bean leaves. The capacity of fungicides applied to the target leaf to redistribute in secondary splash droplets was examined in terms of the infectivity of the spores in the droplets. The extent to which a copper fungicide reduced infection on the receptor leaves was related to the level and tenacity of the fungicide deposit on the target leaf. The effect of wetting agents on the redistribution of this fungicide could probably be explained by their influence on the tenacity of the initial deposit. In general the capacity of different fungicides to inhibit infection by the secondary droplets was related to the inherent toxicity of the fungicides to B. fabae. Implications of the dispersal of spores and fungicides by rain splash are briefly considered with reference to field conditions.     

结合光谱法在DMIs类杀真菌剂筛选中的应用

为建立一种快捷和准确的方法用于新型杀真菌剂的筛选,以外源表达的稻瘟菌羊毛甾醇14α-去甲基化酶为靶酶,以市售烯唑醇、戊唑醇、三唑醇、三唑酮为DMIs类杀真菌剂代表,分析了靶酶活性、靶酶纯度和靶酶浓度对二者结合光谱的影响,并与生物测试结果比较分析其可靠性。结果表明靶酶的高活性、无其他P450干扰和合适的靶酶浓度是获得准确结合光谱的必要条件。烯唑醇、戊唑醇、三唑醇、三唑酮与靶酶结合常数(Kd)分别为0.143μmol/L、0.24μmol/L、0.257μmol/L、0.307μmol/L,该结果与其对稻瘟菌生长抑制能力(120h-EC50)显著相关,证明结合光谱法可作为一种简便可靠的DMIs类杀真菌剂筛选方法。     

Systemic fungicides reduce the abundance of yeast-like symbiotes and the performance of the planthopper Delphacodes kuscheli (Hemiptera: Delphacidae)

Delphacodes kuscheli is the main vector of Mal de Río Cuarto, the most important viral disease of maize in Argentina. This planthopper harbours obligate fungal mutualists, known as yeast-like symbiotes (YLS), which play a key role in the host life cycle. Here, we evaluated the efficacy of six systemic fungicides, prothioconazole (P), prothioconazole & trifloxystrobin (PT), pyraclostrobin & epoxiconazole (PE), pyraclostrobin, epoxiconazole & fluxapyroxad (PEF), picoxystrobin & ciproconazole (PC), and tebuconazole (T) on the reduction of the abundance of YLS in nymphs and newly emerged adults, and the impact of such reduction on the vector performance. All fungicides, except PE, reduced the number of YLS in nymphs treated from 3rd instar five days after starting treatments. When 3rd instar nymphs were allowed to reach adulthood, the fungicides P, PT and PC caused the highest nymphal mortality, significantly reduced the number of YLS in newly emerged adults, lengthened development time and negatively affected morphometric variables of females. There was also a trend towards a higher occurrence of brachypters. In females, the fungicides P, PT and PC caused a greater reduction of YLS when nymphs were treated to these fungicides from 3rd instar compared with nymphs treated from 4th instar. In males, the YLS number was significantly less when nymphs were fed on plants treated with P, PT, PC, and T in 3rd instar. This showed that fungicide treatments caused a greater reduction of YLS when they were applied in early stages of development. Our results provide an effective method to reduce the abundance of YLS in D. kuscheli, contributing to understand the ecological role that these symbionts could be playing in the success of this maize planthopper pest.     

Signum, a new fungicide with interesting properties in resistance management of fungal diseases in strawberries

Signum, a new fungicide developed by BASF, was applied during 6 successive years against fungal diseases in strawberries. The product is formulated as a water dispersible granule, containing 6.7 % pyraclostrobin and 26.7 % boscalid. Pyraclostrobin is similar in chemistry to other strobilurin fungicides like kresoxim-methyl and trifloxystrobin, registered for fruit disease control. Boscalid belongs to the class of carboxyanilides. Both components in the premix formulation combine two different biochemical modes of action in the fungal cell respiration. Therefore, this co-formulation gives a broad-spectrum activity and also a reduced resistance risk for different target pathogens. Botrytis cinerea is the most important disease on strawberry-fruits and thus the control of fruit rot is mainly focused on this fungus. In average over 6 years, Signum has not only given a very good control against Botrytis fruit rot, but it has also shown a high performance in the control of Colletotrichum. Besides, Signum provides good control of powdery mildew (Podosphaera aphanis) and limits the shift to other fruit rots like leather rot (Phytophthora cactorum and leak (Rhizopus, Mucor). The availability of several categories of fungicide families with a different mode of action gives opportunities in alternating different fungicides and is the best guarantee for a sustainable control of fruit rot in all kinds of strawberry production methods. Signum should be integrated in an overall disease management program. Trials, in which the applications of Signum were timed on disease forecasting, based on environmental factors favorable for Botrytis development, were very promising. This tool can also help in establishing the IPM-concept in the production of strawberries.     

Effects of fungicides on a mycophagous coccinellid may represent integration failure in disease management

The adults and larvae of halyziine coccinellids (Coleoptera: Coccinellidae: Halyziini) are obligate mycophages on hyphae and conidia of powdery mildew (PM) (Erysiphales) fungi, that are plant pathogens warranting chemical control in many managed systems. These insects have been observed to reduce PM severity through consumption. Fungicide applications, however, may interfere with this ecological service. Five commercial fungicides were topically applied to the mycophagous coccinellid, Psyllobora vigintimaculata, in the laboratory to gauge contact toxicity. In order to detect interference in the field, population density of naturally occurring P. vigintimaculata was assessed weekly in a northern California vineyard (Vitis vinifera, cultivar “Chardonnay”) over 3 years in relation to PM (Erysiphe necator) severity and in the presence of various fungicides. Wettable sulfur was toxic to adults in the laboratory, resulting in complete cohort mortality 24 h after application. Topical applications of a strobilurin fungicide (trifloxystrobin) and a demethylation inhibitor fungicide (myclobutanil) also resulted in significant adult mortality. Rapid and complete larval mortality was observed in the laboratory after contact with wettable sulfur and myclobutanil. There was no effect on survival after contact with the PM-antagonistic bacterium, Bacillus subtilis. Vineyard density of P. vigintimaculata was reduced in vines receiving applications of sulfur and myclobutanil, even when considering the covariate PM severity. The microbial antagonist, Streptomyces lydicus, did not significantly affect insect density. Our study questions the integration of chemical disease management with naturally occurring mycophagous agents in some agricultural systems.     

Sensitivity of Venturia inaequalis to Trifloxystrobin and Difenoconazole in Uruguay

The sensitivity of Venturia inaequalis to trifloxystrobin and difenoconazole was studied in Uruguay. Populations of V. inaequalis were collected from apple orchards with different histories of trifloxystrobin use. Sensitivity of populations to trifloxystrobin was analysed using a method for testing spore germination published by FRAC, using a discriminatory concentration of 2.0 mg a.i./l. Resistance to trifloxystrobin was widespread in the region of commercial apple production with resistance detected in all orchards examined, the incidences ranging from 3% to 95%. The highest frequencies were found in orchards with the most intensive use of Quinone outside inhibitors (Q_oI) fungicides. Sensitivities of isolates of V. inaequalis to difenoconazole were assessed at five concentrations using a mycelial growth assay on isolates (33 isolates per orchard) from one non‐commercial (baseline orchard) and two commercial orchards having differing histories of difenoconazole use. Populations in both commercial orchards exhibited reduced sensitivities to difenoconazole compared to the baseline orchard. Resistance factor (RF) values of 6.6 and 11.74 were measured in the orchards with moderate (up to 4 sprays per season) and intensive use (more than 5 sprays per season) of difenoconazole, respectively. A single‐assessment concentration (SAC) was identified for assessing difenoconazole sensitivity of V. inaequalis isolates by fitting linear regressions between log₁₀ EC₅₀ and relative growth (RG) of the isolates at each fungicide concentration testing, and comparing the goodness‐of‐fit of the regression lines. Comparable results were obtained based on EC₅₀ values and RG values at a SAC of 0.05 mg of active ingredient per litre (a.i. per l). Populations from both commercial orchards differed from the baseline population, in that isolates with RG ≥70 were present at substantial levels in the former but absent from the latter.