Agronomic strategies to enhance the early vigor and yield of maize part II: the role of seed applied biostimulant, hybrid, and starter fertilization on crop performance.

Maize cropping systems need to be re-designed, within a sustainable intensification context, by focusing on the application of high-use efficiency crop practices, such as those that are able to enhance an early plant vigor in the first critical growth stages; such practices could lead to significant agronomic and yield benefits. The aim of this study has been to evaluate the effects of the cultivation of hybrids with superior early vigor, of the distribution of starter fertilizers at sowing, and of the seed application of biostimulants on promoting plant growth and grain yield in full factorial experiments carried out in both a growth chamber and in open fields. The greatest benefits, in terms of plant growth enhancement (plant height, biomass, leaf area) and cold stress mitigation, were detected for the starter fertilization, followed by the use of an early vigor hybrid and a biostimulant seed treatment. The starter fertilization and the early vigor hybrid led to earlier flowering dates, that is, of 2.1 and 2.8 days, respectively, and significantly reduced grain moisture at harvest. Moreover, the early vigor hybrid, the starter NP fertilization, and the biostimulant treatment increased grain yield by 8.5%, 6.0%, and 5.1%, respectively, compared to the standard hybrid and the untreated controls. The combination of all the considered factors resulted in the maximum benefits, compared to the control cropping system, with an increase in the plant growth of 124%, a reduction of the sowing-flowering period of 5 days, and a gain in grain yield of 14%. When choosing the most suitable crop practice, the diversity of each cropping system should be considered, according to the pedo-climatic conditions, the agronomic background, the yield potential, and the supply chain requirements.

Multi-Mycotoxin Long-Term Monitoring Survey on North-Italian Maize over an 11-Year Period (2011-2021): The Co-Occurrence of Regulated, Masked and Emerging Mycotoxins and Fungal Metabolites.

Maize is considered one of the most susceptible crops to mycotoxin-producing fungi throughout the world, mainly belonging to the Fusarium spp. and Aspergillus spp. Maize is mainly used as animal feeds in Italy, as well as for human consumption, being essential for all the protected designation of origin (DOP) products. Our study investigated the occurrence of regulated mycotoxins in 3769 maize grain samples collected from 88 storage centers by the National Monitoring Network over an 11-year period (2011-2021). Moreover, an in-depth survey over a 4-year period, characterized by extremely different meteorological conditions, was conducted to investigate the co-occurrence of regulated, masked, and emerging mycotoxins. The survey confirmed that Fusarium spp. was the most frequent fungi and fumonisins were the main mycotoxins that were constantly detected in the different years and areas. Moreover, the areas characterized by high fumonisin levels were also the most prone to contamination by emerging mycotoxins produced by the same Fusarium species of the Liseola section. On the other hand, as a result of climatic changes, maize grains have also been affected by the increased frequency of aflatoxin accumulation. Deoxynivalenol, zearalenone, and other emerging mycotoxins produced by the same Fusarium species as the Discolor section occurred more abundantly in some areas in Northern Italy and in years characterized by predisposing meteorological conditions.

The Effect of Foliar Fungicide and Insecticide Application on the Contamination of Fumonisins, Moniliformin and Deoxynivalenol in Maize Used for Food Purposes.

The fungal ear rot of maize cultivated in temperate areas is mainly due to the Fusarium species. The use of insecticides against European Corn Borer (ECB) reduces the severity of fungal ear rot as well as the fumonisin (FB) and moniliformin (MON) levels in maize kernels at harvest, which in turn results in a lowering of their effect on deoxynivalenol (DON) control. However, the direct fungicidal control of ear rot has rarely been implemented for maize, and the first studies reported conflicting results on the reduction of mycotoxins. In the present experiment, field trials were carried out in North Italy over three growing seasons to study the effect of fungicide application timings on maize to control mycotoxins, considering the interaction of the application with the insecticide treatment, according to a full factorial split plot design. The mycotoxin content was determined through LC-MS/MS analysis. The field trials showed a significant reduction in ECB severity (75%), fungal ear rot severity (68%), Fusarium Liseola section infection (46%), FBs (75%) and MON (79%) as a result of the insecticide application for all the years, while the DON content increased by 60%. On the other hand, a fungicide application alone or applied in plots protected by an insecticide was never effective for the fungal symptoms, infection or mycotoxin content. The results confirm that a correct insecticide application to control ECB damage is the most effective agrochemical solution for the control of fungal ear rot, FBs and MON.

The Role of Nitrogen Fertilization on the Occurrence of Regulated, Modified and Emerging Mycotoxins and Fungal Metabolites in Maize Kernels.

The European Food Safety Authority is currently evaluating the risks related to the presence of emerging mycotoxins in food and feeds. The aim of this study was to investigate the role of soil fertility, resulting from different nitrogen fertilization rates, on the contamination of regulated mycotoxins and emerging fungal metabolites in maize grains. The trial was carried out in the 2012-2013 growing seasons as part of a long-term (20-year) experimental platform area in North-West Italy, where five different N rates, ranging from 0 to 400 kg N ha-1, were applied to maize each year. Maize samples were analyzed by means of a dilute-and-shoot multi-mycotoxin LC-MS/MS method, and more than 25 of the most abundant mycotoxins and fungal metabolites were detected. Contamination by fumonisins and other fungal metabolites produced by Fusarium spp. of the section Liseola was observed to have increased in soils that showed a poor fertility status. On the other hand, an overload of nitrogen fertilization was generally associated with higher deoxynivalenol and zearalenone contamination in maize kernels, as well as a higher risk of other fungal metabolites produced by Fusarium spp. sections Discolor and Roseum. A balanced application of N fertilizer, in accordance with maize uptake, generally appears to be the best solution to guarantee an overall lower contamination by regulated mycotoxins and emerging fungal metabolites.

Characterization and selection of functional yeast strains during sourdough fermentation of different cereal wholegrain flours.

The increasing demand for healthy baked goods boosted studies on sourdough microbiota with beneficial metabolic traits, to be used as potential functional starters. Here, 139 yeasts isolated from cereal-based fermented foods were in vitro characterized for their phytase and antioxidant activities. The molecular characterization at strain level of the best 39 performing isolates showed that they did not derive from cross contamination by baker's yeast. Afterwards, the 39 isolates were in vivo analyzed for their leavening ability, phytase activity and polyphenols content using five different wholegrain flours, obtained from conventional and pigmented common wheat, emmer and hull-less barley. Combining these findings, through multivariate permutation analysis, we identified the 2 best performing strains, which resulted diverse for each flour. Doughs singly inoculated with the selected strains were further analyzed for their antioxidant capacity, phenolic acids, xanthophylls and anthocyanins content. All the selected yeasts significantly increased the total antioxidant activity, the soluble, free and conjugated, forms of phenolic acids and anthocyanins of fermented doughs. This study revealed the importance of a specific selection of yeast strains for wholegrain flours obtained from different cereals or cultivars, in order to enhance the pro-technological, nutritional and nutraceutical traits of fermented doughs.

Distribution of bioactive compounds in pearled fractions of tritordeum.

Hexaploid tritordeum is the amphidiploid cereal derived from the cross between wild barley and durum wheat. The present study compares two cultivars of tritordeum with other cereals grown in the same experimental area to weigh up its potential use as ingredient for health-valued foods. Tritordeum shows 2.5-fold higher concentration of lutein than common wheat and barley, and 1.2-fold higher than durum wheat, while the concentration of ß-glucans is 5 folds lower than the one observed for barley. Based on the distribution of bioactive compounds in pearled fractions, the use of whole-grain flours seems the best way to exploit the antioxidant potential of tritordeum. Nevertheless, the internal layers of the kernel of this cereal are characterized on average by high concentrations of antioxidants (32.0 mg/kg and 518 mg/kg soluble and cell wall-bound phenolic acids, respectively), making tritordeum interesting also for the production of refined flours rich in bioactive compounds.

Development and Comparison of Two Multiresidue Methods for the Determination of 17 Aspergillus and Fusarium Mycotoxins in Cereals Using HPLC-ESI-TQ-MS/MS.

Cereals can be contaminated by several mycotoxins, whose co-presence may represent an undervalued risk for humans and animals. Maize and wheat are the most contaminated cereals and in temperate areas could be affected in field conditions by several Fusarium and Aspergillus infections. To date, only B-fumonisins (FBs), aflatoxins (AFs), zearalenone (ZEA), deoxynivalenol (DON) and T-2 and HT-2 toxins have been regulated in cereals in European Union. The other fungal metabolites, are commonly referred to as "emerging" and "masked" mycotoxins, and more information on their occurrence in combination with the regulated mycotoxins, are needed to design combined toxicological and exposure assessments. This research intends to develop and compare two multiresidue HPLC-ESI-TQ-MS/MS methods for the simultaneous determination of the main regulated, emerging and masked mycotoxins in maize and wheat, among which: FB1, FB2, DON, ZEA, AFB1, AFB2, AFG1, AFG2, moniliformin (MON), deoxynivalenol-3-glucoside (DON-3-G), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), nivalenol (NIV), enniatins A, A1, B, B1 (ENNA, ENNA1, ENNB, ENNB1). The extraction was performed for both methods using a mixture of CH3CN/H2O/CH3COOH (79/20/1, v/v/v), while the dilution/purification was carried out through two different procedures: (1) by the "dilute-and-shoot" technique diluting 1:2 the filtered extract with CH3CN/H2O/CH3COOH (20/79/1, v/v/v) to reduce the matrix effect; (2) using the Oasis® PRiME HLB clean-up columns. The analysis was carried out using CH3OH and H2O both acidified with 0.1% of CH3COOH as eluents. The injection volume was 20 µL and the flow rate 200 µL min-1. The analysis of two reference material (maize and wheat), was performed to evaluate the trueness and precision of the two methods by matrix-matched calibration curves. For all the regulated mycotoxins analyzed by both methods, the range of recovery percentage established by the Regulation (EC) No. 401/2006 was respected, except for ZEA by using the Oasis® PRiME HLB clean-up columns. Nevertheless, the results suggest that the Oasis® PRiME HLB clean-up columns, could be a valid alternative to the dilute-and-shoot method, although an additional cost for the clean-up has to be considered. In conclusion, both two analytical methods considerably reduce the analytical time and costs and therefore result to be promising and applicable for high-throughput routine multi-mycotoxins analysis by the use of a TQ.

Bioactive compound and antioxidant activity distribution in roller-milled and pearled fractions of conventional and pigmented wheat varieties.

In this study, the chemical composition of pigmented wheats (yellow, purple and blue types), and the distribution of the bioactive compounds in their roller-milled and pearled fractions, were compared with conventional wheats (red and white types). Roller-milling promoted the recovery of total dietary fiber, ß-glucans, phenolic acids and anthocyanins in the bran fraction, which resulted also in a higher total antioxidant activity than the refined flour. Conversely, lutein resulted mainly concentrated in the refined flour. In the same way, the distribution pattern in the pearled fractions differ depending on the bioactive considered. The study highlights that a careful selection of the most appropriate fractionation process should be performed to produce flours rich in bioactive compounds. Roller-milling resulted useful for the production of refined flours rich in xanthophylls, with particular emphasis to the yellow-grained wheats. Contrarily, pearling could be more useful in the valorization of the health potential of anthocyanin-pigmented varieties.

Folate distribution in barley (Hordeum vulgare L.), common wheat (Triticum aestivum L.) and durum wheat (Triticum turgidum durum Desf.) pearled fractions.

BACKGROUND: Wholegrain cereals are an important source of folates. In this study, total folate was analysed in pearled fractions of barley and wheat cultivars employing AOAC Official Method 2004.05. In particular, the distribution of folate in the kernels was evaluated in three barley cultivars (two hulled types and a hulless one as well as two- and six-row types) and in a common and a durum wheat cultivar. RESULTS: A noticeable variation in the folate content was observed between the barley [653-1033 ng g(-1) dry matter (DM)] and wheat cultivars (1024-1119 ng g(-1) DM). The highest folate content was detected in the hulless barley cultivar (1033 ng g(-1) DM). A significant reduction in total folate, from 63% to 86%, was observed in all cultivars from the outermost to the innermost pearled fractions. CONCLUSION: Results proved that folates are mainly present in the germ and in the outer layers of the kernel. This is the first study reporting the folate distribution in kernels of both common and durum wheat and in a hulless barley cultivar. Results suggest that the pearling process could be useful for the selection of intermediate fractions that could be used in order to develop folate-enhanced ingredients and products.

Progressive Pearling of Barley Kernel: Chemical Characterization of Pearling Fractions and Effect of Their Inclusion on the Nutritional and Technological Properties of Wheat Bread.

Two hulled barley varieties have been sequentially pearled for one to eight cycles, each with 5% removal. The derived fractions were analyzed for their bioactive compound content. The dietary fiber (DF) decreased from the external to the internal layers, whereas ß-glucans showed an inverse trend. Deoxynivalenol contamination was concentrated in the outer layers. The total antioxidant activity (TAA) was higher in the 15-25% fractions, which were used to prepare bread. Five mixtures of refined wheat flour, with an increasing replacement of this pearled barley fraction, were compared with a control for the bioactive compound content, as well as for the rheological and physical bread properties. The inclusion of pearled fractions with up to a 10% substitution leads to a clear enhancement of the DF and TAA, with only minor detrimental effects on the physical parameters. Selected byproducts of barley pearling could be proposed as functional ingredients for bakery products rich in DF and TAA.

Role of the European corn borer (Ostrinia nubilalis) on contamination of maize with 13 Fusarium mycotoxins.

The European corn borer (ECB) plays an important role in promoting Fusarium verticillioides infections and in the consequent fumonisin contamination in maize grain in temperate areas. The objective of this study was to evaluate whether the ECB feeding activity could also affect the occurrence of emerging mycotoxins in maize kernels. During the 2008-10 period, natural infestation of the insect was compared, in field research, with the protection of infestation, which was obtained by using an entomological net. The ears collected in the protected plots were free from ECB attack, while those subject to natural insect attacks showed a damage severity that varied from 10% to 25%. The maize samples were analysed by means of an LC-MS/MS-based multi-mycotoxin method, which led to the detection of various metabolites: fumonisins (FUMs), fusaproliferin (FUS), moniliformin (MON), bikaverin (BIK), beauvericin (BEA), fusaric acid (FA), equisetin (EQU), deoxynivalenol (DON), deoxynivalenol-3-glucoside (DON-3-G), zearalenone (ZEA), culmorin (CULM), aurofusarin (AUR) and butenolide (BUT). The occurrence of mycotoxins produced by Fusarium spp. of Liseola section was affected significantly by the ECB feeding activity. The presence of ECB injuries increased the FUMs from 995 to 4694 µg kg(-1), FUS from 17 to 1089 µg kg(-1), MON from 22 to 673 µg kg(-1), BIK from 58 to 377 µg kg(-1), BEA from 6 to 177 µg kg(-1), and FA from 21 to 379 µg kg(-1). EQU, produced by F. equiseti section Gibbosum, was also increased by the ECB activity, by 1-30 µg kg(-1) on average. Instead, the content of mycotoxins produced by Fusarium spp. of Discolor and Roseum sections was not significantly affected by ECB activity. As for FUMs, the application of a strategy that can reduce ECB damage could also be the most effective solution to minimise the other mycotoxins produced by Fusarium spp. of Liseola section.

Nutritional and technological quality of bread enriched with an intermediated pearled wheat fraction.

A strategy to maximise the health benefits of wheat-based products enriched with refined flour and selected fractions of kernel, obtained by sequential pearling, has been tested. Five mixtures of refined commercial flour with an increasing replacement of a pearled wheat fraction were used to prepare bread and were compared with a control for the dough rheological properties (Mixolab® parameters), the bioactive compound content, deoxynivalenol (DON) contamination and the physical properties (volume, crust colour, instrumental crunchiness and crumb texture profile analysis parameters). The antioxidant and dietary fibre contents increased linearly as the flour was enriched with the pearled fraction. The dietary fibre, ß-glucan, total phenolic, alkylresorcinol content and the antioxidant activity increased significantly at a replacement level of 10%, while the technological properties were not significantly different from those of the control. It has been shown that refined flour could be enriched through the addition of a selected wheat pearled fraction and the bioactive compound content of composite bread could be improved, while few rheological and technological differences could be obtained and the risk for DON contamination could be limited.

Moniliformin analysis in maize samples from North-West Italy using multifunctional clean-up columns and the LC-MS/MS detection method.

A fast clean-up method has been developed to purify maize extracts and to detect moniliformin (MON) in maize samples from North-West Italy over a four-year period (2008-2011). The method is based on the use of MycoSep® 240 Mon clean-up columns (Romer Labs®). Samples were extracted using acetonitrile/water (84:16, v/v), and the extracts were purified with previously described clean-up columns. The liquid chromatography-tandem mass spectrography (LC-MS/MS) analysis has been carried out by means of hydrophilic interaction chromatography (HILIC), combined with negative electrospray mass spectrometry. The method has a recovery of 76-91% (relative standard deviation, RSD%: 6-14%), a limit of detection (LOD) of 1 µg kg⁻¹ and a limit of quantification (LOQ) of 4 µg kg⁻¹. Naturally contaminated maize (108 samples) was analysed for MON content. The average percentages of positive samples was 93% with the following ranges (µg kg⁻¹): 33-2606 (2008);

Organically modified clays as binders of fumonisins in feedstocks.

This study reports an investigation on the ability of organically modified clays to bind mycotoxins, fumonisins B1 (FB1) and B2 (FB2). Organically modified clays are commercia materials prepared from natural clays, generally montmorillonite, by exchanging the inorganic cation with an ammonium organic cation. A screening experiment conducted on 13 organically modified clays and 3 nonmodified clays, used as controls, has confirmed that the presence of an organic cation in the clay interlayer promoted the adsorption of both fumonisins. On the basis of the results of the screening test, four modified clays and a Na-montmorillonite were selected for the determination of the adsorption kinetics and isotherms. On all the tested materials adsorption took place within one hour of contact with fumonisins solutions. Adsorption isotherms have pointed out that the modified clays exhibited a higher adsorptive capacity than the unmodified clay. It was also demonstrated that, notwithstanding the reduced structural difference between FB1 and FB2, they were differently adsorbed on the modified clays. Addition of 2% modified clays to contaminated maize allowed a reduction of more than 70% and 60% of the amount of FB1and FB2 released in solution. Although in vivo experiments are required to confirm the effectiveness of the organically modified clays, these preliminary results suggest that these materials are promising as fumonisins binders.

Prediction of the dry-milling performance of maize hybrids through hardness-associated properties.

BACKGROUND: The hardness of kernels determines the dry-milling processing performance of maize hybrids. The identification of the best maize hybrids for the dry-milling process requires a limited number of simple, practical and reliable tests that are able to predict the potential grit yield. RESULTS: A total of 119 samples from different genetic and environmental backgrounds, collected over 3 years, were analysed for coarse/fine ratio (C/F), floating test (FLT), protein content (PC), kernel sphericity (S), total milling energy (TME) and test weight (TW). The total grit yield (TGY) of each sample was obtained through a micromilling procedure based on the manual separation of kernel endosperm followed by grinding and sieving under standard operational conditions. The TGY was used to establish the capability of the tests to predict the dry-milling aptitude. Single and multiple linear regression analyses were performed to establish equations for the prediction of TGY using C/F, FLT, PC, S, TME and TW as independent variables. The analyses were performed on three data sets, clustered year by year of the sample collection and analysis, and the resulting average coefficients of determination (R(2)) were compared by analysis of variance. C/F, FLT, TME and, to a lesser extent, TW appeared to be easy-to-use independent descriptors of maize dry-milling. An improved model prediction ability was observed when different combinations of a few physical and chemical properties were used as input variables. However, the models in which three or more variables were used did not lead to any significant improvement in TGY prediction compared with the smaller models. CONCLUSION: This study contributes towards establishing the best predictor of maize kernel aptitude to dry-milling processes. Of all considered data sets, a milling evaluation factor (C/F or TME) coupled with kernel density (measured by means of the FLT) showed the best predictive ability for dry-milled product yields.

Determination of maize kernel hardness: comparison of different laboratory tests to predict dry-milling performance.

BACKGROUND: Numerous foods are produced from maize, and grain hardness has been described to have an impact on grain end-use value, and in particular for dry-milling performance. RESULTS: Thirty-three samples of commercial hybrids have been analysed for test weight (TW), thousand-kernel weight (TKW), hard:soft endosperm ratio (H/S), milling time (MT) and total milling energy (TME) through the Stenvert hardness test, coarse:fine material ratio (C/F), break force (HF) and break energy (HWF) through the puncture test, floating test (FLT), kernel dimensions and sphericity (S), protein (PC), starch (SC), lipid (LC), ash (AC) content and amylose:amylopectin ratio (AS/AP).Total grit yield (TGY) has been obtained through a micromilling procedure and used to compare the efficiency of the tests to predict the dry-milling performance. TW, H/S, MT, TME, C/F, FLT, S, PC, SC and AS/AP were significantly correlated with each other. TW has been confirmed to be a simple estimator of grain hardness. Among the hardness tests, C/F was shown to be the best descriptor of maize milling ability, followed by FLT. A good correlation with TGY has also been observed with H/S, MT, TME and PC, while SC, S and AS/AP seem to play a minor role. The puncture test (HF and HWF) did not offer good indications on the impact of hardness on kernel grinding properties. CONCLUSION: This study can be considered as a contribution towards determining kernel properties which influence maize hardness measurement in relation to the end-use processing performance.

Timing clorpirifos + cypermethrin and indoxacarb applications to control European corn borer damage and fumonisin contamination in maize kernels.

BACKGROUND: European corn borer (ECB) is the main maize pest in central and southern Europe and it promotes the infection of maize with Fusarium verticillioides, which is able to produce fumonisins. Field experiments were performed from 2006 to 2007 in northwestern Italy to determine the effects of the timing of insecticide applications on maize fungal ear rot and fumonisin contamination in natural infection conditions. Four application timings and two insecticides (clorpirifos + cypermethrin and indoxacarb) were compared each year. RESULTS: In both years, the treatments applied at the beginning of a consistent ECB flight activity and at the flight peak showed the best efficacy to control the insect damage on ears. Fungal ear rot and fumonisin contamination were clearly affected by ECB control. The efficacy of the best application timing to control fumonisin occurrence was 73% in 2006 and 84% in 2007. Earlier insecticide applications showed lower fumonisin contamination than treatments applied after the adult flight peak. CONCLUSION: The production of maize and maize-based foods with a low fumonisin content may be enhanced through correct insecticide application against the second ECB generation. The optimum timing of insecticides is between the beginning of a consistent adult flight activity and the flight peak.

Management of fumonisin contamination in maize kernels through the timing of insecticide application against the European corn borer Ostrinia nubilalis Hübner.

The European corn borer (ECB), Ostrinia nubilalis, is the principal pest of maize in Central and South Europe. It is known to promote Fusarium verticillioides infection in maize grain, a recognized producer of fumonisins. Field experiments were performed in 2006 and 2007 in two sites in NW Italy to determine the effects of the timing of insecticide application on ECB damage, fungal ear rot and fumonisin contamination under natural conditions. Different insecticide application timings were compared, from maize flowering to approximately 15 days after the flight peak of adult ECB. At harvest, the ears were rated for incidence and severity of ECB damage, fungal ear rot symptoms and fumonisin (FB(1) + FB(2)) contamination. In all years/sites, treatments applied at the beginning of consistent ECB flight activity were most effective in controlling insect damage on ears. Fungal ear rot and fumonisin contamination were significantly affected by ECB control. The efficacy of the best timing of insecticide application in controlling fumonisin contamination was, on average, 93% compared to the untreated control. Contamination levels of these mycotoxins increased with either an earlier or later treatment. Furthermore, an earlier insecticide application showed lower fumonisin contamination than a treatment applied after the adult flight peak. Production of maize kernels and maize-based foods that do not exceed the maximum international and EU permitted levels for fumonisins could be enhanced by appropriate insecticide treatment against second generation ECB. The optimum time for insecticide application is between the beginning of consistent adult flight activity and the flight peak.

Impact of insecticide treatments on Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) and their influence on the mycotoxin contamination of maize kernels.

BACKGROUND: European corn borer (ECB), Ostrinia nubilalis (Hübner), is the main maize pest in Central and Southern Europe and promotes the infection of maize with Fusarium spp., which produce mycotoxins. The objective of this study was to determine the effect of insecticide treatments on ECB damage, Fusarium infection and mycotoxin contamination. The field experiments were performed from 2000 to 2006 in three locations in north-western Italy. At harvest, ears were rated for the incidence and severity of ECB damage and Fusarium ear rot symptoms, and the harvested kernels were analysed for fumonisins and zearalenone. RESULTS: In all the years except 2003, the chemical treatment had a significant effect on ECB incidence and severity. The ear damage was reduced, on average, by 44.1%. Fusarium ear rot and fumonisin contamination were affected by ECB control. The occurrence of the mycotoxin was significantly reduced, on average by 68%. The differences between the treatments were less clear in the year with highest ECB pressure. No significant difference was observed between the different insecticide classes. No relations were detected between the ECB activity and the occurrence of zearalenone. CONCLUSION: This research indicates that the production of kernels with low fumonisin content may be enhanced by an insecticide treatment against the second ECB generation.