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Due to climate changes occurrence of Fusarium fujikuroi species complex (FFSC) on small grain has been observed in the last decade in higher frequency. A significant aspect of the FFSC is their ability to synthesize mycotoxins which can have a harmful effect on the health of people and domestic animals, which is reflected in the manifestation of carcinogenic, teratogenic, neurotoxic and other toxic effects. The aim of this study was to examine the virulence of FFSC isolates and their ability to synthesis fumonisins. Total of 50 FFSC isolates originating from 17 localities in Serbia and isolated from 4 plant hosts (wheat, durum wheat, barley and triticale) were collected from MRI Collection of fungi. The virulence of the isolates was tested on durum wheat spikes in a two-year study. The concentrations of synthesized fumonisins were tested using the ELISA method. Significant difference was recorded between the two years of research, in the sense that in the second year all isolates were more virulent. Regarding the influence of the species on virulence, it was shown that the species F. verticillioides is more virulent than the species F. proliferatum and F. subglutinans, which are equally virulent in both years. The results of the fumonisin synthesis potential indicated the presence of fumonisin in all isolates of the species F. verticillioides and F. proliferatum in high concentrations. Fumonisin synthesis was recorded only in some isolates of the species F. subglutinans while in the other isolates it was not detected.

Maize contamination is a global concern due to its critical role in the food and feed supply chain and its susceptibility to aflatoxin contamination. There is heightened attention on aflatoxins because they reduce agricultural yields, leading to substantial global economic losses, and pose threats to food safety owing to their highly toxic and carcinogenic properties. Aspergillus parasiticus exists in both virulent and non-virulent strains, which, under varying climatic conditions, can produce specific aflatoxins, with aflatoxin B1 (AFB1) being the most carcinogenic. Early detection of toxigenic fungi, coupled with accurate species identification, is critical for implementing an effective strategy to minimize fungal growth and mycotoxin production. This approach aims to safeguard maize cultivation and its final products. Because each toxigenic fungal species has its unique mycotoxin profile, accurate species identification is crucial for effective mycotoxin prevention strategies. Identifying species based on morphological characteristics is time-consuming and demands expert taxonomists with specialized knowledge in specific groups of species. In presented work species-specific primers were tested for rapid, sensitive, simultaneous, and PCR-based identification of fungal species. We have verified the presence of A. parasiticus using the specific primers (AFLA1/AFLA2) developed by Susca et al. in 2020. In all 20 isolates previously identified using a multidisciplinary approach, the presence of this species was confirmed. Since this species synthesizes aflatoxins, rapid and timely identification is essential.

Species of the genus Aspergillus are distributed throughout the world but are primarily represented in subtropical and warm temperate climates. In warm, humid subtropical or tropical climates or even in temperate climates in which the growing season is unusually hot and dry, species of the genus Aspergillus can infect seeds in the fields. The movement of fungal species threatens food security and creates global public concern worldwide. Climate change is altering the distribution, incidence and intensity of plant pests that is moving north into more temperate zones of Europe. In recent years, Serbia has been facing climate changes accompanied by a warming trend, which is a cause for concern. A more frequent occurrence of Aspergillus species was observed in the years in which the weather conditions were characterized by high temperatures, lack of precipitation and pronounced drought conditions (2012, 2015, 2017). The intensive outbreaks of Aspergillus species were in 2012. In addition to the previously known species Aspergillus flavus, which was a storage pathogen, it also occurs in field conditions. We discovered a new species of A. parasiticus in Serbia, for the first time. During the 2015 and 2017 we identified new species of the genus Aspergillus section Nigri. In order to adapt to climate change, priority is given to the cultivation of drought-resistant maize hybrids. The introduced challenges require additional climate change adaptation strategies, including various changes in production systems, in order to maintain long-term productivity.

Wheat and wheat products can be contaminated with various fungal species. Secondary metabolites of these fungi, such as deoxynivalenol, zearalenone, fumonisin B1 and aflatoxins are important mycotoxins that cause various harmful effects on human and animal health. Therefore, the aim of this study was to examine the concentration of mycotoxins in wheat grain. Wheat samples were tested for the presence of four mycotoxins (fumonisins, aflatoxins, deoxynivalenol and zearalenone). Samples were collected during two years (2020-2021), from different plots in Zemun Polje, Serbia. Grain samples were selected randomly. Total of 10 samples were tested each year. In order to perform the analysis for the presence of mycotoxins, grains were dried and ground. The analysis was performed by ELISA (Enzyme-linked immunosorbent assay) using Tecna´s ELISA test kit for mycotoxins. During the 2020 no fumonisins were recorded in any of the wheat samples. The values of synthesized aflatoxins ranged from 0.618-1.818 ppb, the values of deoxynivalenol ranged from 0.006-1.058 ppm while concentrations of zearalenone were highest and ranged from 0.230-9.379 ppb. In 2021 no fumonisins and zearalenone were recorded in any of the wheat samples. Aflatoxins ranged from 2.209-3.812 ppb, and deoxynivalenol ranged from 1.244-3.307 ppm. Contamination of wheat grain with toxic metabolites of fungi is one of the particularly important problems in global agriculture because it causes significant economic losses. Therefore, it is important to monitor the presence of these metabolites in order to develop effective methods to reduce grain contamination.

Ova studija je sprovedena sa ciljem da se ispita prirodna pojava toksigenih vrsta gljiva i sadržaj fumonizina (FB), ukupnih aflatoksina (AFLA), zearalenona (ZEA) i deoksinivalenola (DON) u zrnu kukuruza, uskladištenom neposredno nakon berbe 2022. godine. Prikupljeni su uzorci zrna kukuruza sa tri lokacije (Zemun Polje, Školsko Dobro i Zagajica) u Srbiji i analizirani na prisustvo mikotoksina. Nakon sušenja i mlevenja, uzorci su homogenizovani sa 25 ml 70% rastvora metanola i destilovanom vodom (3:1), a zatim ekstrahovani. Kvantifikacija ukupnog sadržaja mikotoksina izvršena je metodom imunoapsorpcionih enzima (ELISA) prema uputstvu za upotrebu proizvođača (Tecna S.R.L., Italija, Celer Test Kit). Analizom 100 uzoraka zrna kukuruza utvrđena je velika varijabilnost u koncentraciji ispitivanih mikotoksina. Svi ispitivani uzorci su bili pozitivni na najmanje jedan od ispitivanih mikotoksina (FB, AFLA, ZEA, DON). Fumonizin je detektovan u koncentraciji od 0 do 0,268 ppm, ukupni aflatoksin od 0,423 do 3,925 ppb, zearalenon od 0 do 9,685 ppb, i deoksinivalenol od 0,005 do 3,581 ppm. U svim ispitivanim hibridima, analize mikotoksina su pokazale da su nivoi FB, AFLA, ZEA i DON bili ispod maksimalno dozvoljenih nivoa propisanih zakonodavstvom Evropske unije i Republike Srbije, namenjenom za kukuruz i proizvode od kukuruza. Neophodno je kontinuirano praćenje sadržaja mikotoksina, s obzirom da se isti menja iz godine u godinu., This study was conducted to investigate the natural occurrence of toxigenic fungal species and the levels of fumonisin (FB), zearalenone (ZEA), deoxynivalenol (DON), and total aflatoxin (AFLA) in corn kernels stored immediately after harvest in 2022. Samples of maize kernels from two locations (Zemun Polje and Zagajica) in Serbia were collected and analyzed for the presence of mycotoxins. After drying and milling, the samples were homogenized with 25 ml of 70% metanol solution and distilled water (3:1) and then extracted. Quantification of total mycotoxin content was performed using the immunoapsorption enzyme method according to the manufacturer's instructions (Tecna S.R.L., Italy, Celer Test Kit). The analysis of 100 maize kernels samples was determined by a great variability in the concentration of mycotoxins studied. All samples tested were positive for at least one of the mycotoxins tested (fumonisin, aflatoxin, zearalenone or deoxynivalenol). Fumonisin is determined at a concentration of 0 to 0.268 ppm, total aflatoxin at a concentration of 0.423 to 3.925 ppb, zearalenone at a concentration of 0 to 9.685 ppb, while deoxynivalenol is detected at a concentration of 0.005 to 3.581 ppm. In all tested hybrids, mycotoxin analyses showed that the levels of AFLA, DON, ZEA and FBs were below the maximum permissible levels established by the legislation of the European Union and the Republic of Serbia in maize intended for maize and maize products. Continuous monitoring of mycotoxin content is necessary as it changes from year to year.

This study was carried out in order to investigate the natural occurrence of toxigenic fungal species and levels of fumonisin (FBs), zearalenone (ZEA), deoxynivalenol (DON) and total aflatoxin (AFLA) in the maize kernels, stored immediately after harvesting in 2022. Samples of maize kernels from two locations (Zemun Polje and Zagajica) in Serbia were collected, and analysed for the presence of mycotoxins. After drying and grinding, the samples were homogenized with 25 ml of 70% metanol solution and distilled water (3:1), and then extracted. Quantification of the total content of mycotoxins was performed using the immunoapsorption enzyme method according to the manufacturer’s operating instructions (Tecna S.R.L., Italy, Celer Test Kit). The analysis of 100 maize kernels samples was determined by great variability in the concentration of examined mycotoxins. All examined samples were positive for at least one of the examined mycotoxins (fumonisin, aflatoxin, zearalenone or deoxynivalenol). Fumonisin is determined in concentration of 0 to 0.254 ppm, total aflatoxin in concentration of 0.619 to 3.676 ppb, zearalenone in concentration 0 to 9.379 ppb, while deoxynivalenol is detected in concentration 0.006 to 3.307 ppm. In all tested hybrids, mycotoxins analyses showed that the levels of AFLA, DON, ZEA and FBs were below the maximum permissible levels stipulated by the legislation of the European Union and the Republic of Serbia in maize intended for maize and maize products. Continuous monitoring of mycotoxin content is necessary, given that it changes from year to year.

U ovom istraživanju je analizirana učestalost prisustva toksigenih vrsta gljiva na zrnu šest hibrida kukuruza iz različitih FAO grupa zrenja (ZP 341, ZP 427, ZP 434, ZP 560, ZP 606, ZP 666). Uzorci su prikupljeni u vreme berbe, iz tri različita lokaliteta (Zemun Polje, Bečej, Šabac) u Srbiji. Koristeći standardne mikološke metode izolacije, istraživači su izolovali toksigene vrste gljiva. Dobijene kolonije su prečišćene kako bi se dobila čista kultura gljiva. Ove kulture su potom korišćene za identifikaciju različitih vrsta gljiva. Kulture gljiva su gajene na različitim podlogama kao što su krompir-dekstroza podloga (PDA), podloga sa fragmentima sterilnog lista karanfila (CLA) i sintetička podloga (SNA). Inkubacija kultura je obavljena pri kontrolisanoj temperaturi od 25±1ºC. Identifikacija izolovanih vrsta je izvršena prema Nelsonui sar. (1983). Statistička analiza prikupljenih podataka izvedena je korišćenjem analize varijanse (ANOVA), uz primenu paketa Statistica 10 (StatSoft, Inc., SAD). Značajnost razlika između srednjih vrednosti parametara je ocenjena putem F-testa pri nivou značajnosti od P ≤ 0,05. Tokom mikoloških istraživanja zrna kukuruza, potvrđeno je prisustvo toksigenih gljiva iz tri roda: Aspergillus, Fusarium i Penicillium. U lokalitetima koji su obuhvaćeni istraživanjem, gljiva Fusarium verticillioides je bila najčešće izolovana, sa maksimalnom učestalošću od 36% kod većine ispitivanih hibrida. Prisustvo Aspergillus spp. se kretalo od 0 do 19%, dok je prisustvo Penicillium spp. variralo od 0 do 25%. Imajući u vidu povoljne agroekološke uslove u Srbiji za razvoj toksigenih gljiva i njihovih mikotoksina, neophodno je redovno sprovoditi kontrolu nad sadržajem mikotoksina u zrnu kukuruza, kako tokom žetve tako i nakon nje.

During the 2014-2017 period, wheat samples were collected from discoloured spikes affected by Fusarium head blight (FHB) from 20 locations in Serbia. After isolation, fungi were cultivated on potato dextrose agar (PDA) at 25oC for 7 days. Based on the in situ identification on PDA, 36 isolates of the section Liseola were selected for further analyses. The pathogenicity of all isolates was confirmed on wheat leaves. The virulence of isolates was determined by measuring the lengths of spots formed at the inoculation leaf site. In order to prove the presence of the species Fusarium verticillioides, a pair of primers FV-F2/FV-R was used. This pair of primers amplifies the sequence of the gaoB gene, and it proved to be specific for the stated species. Moreover, for the same purpose, a pair of primers VER1-VER2 based on the calmodulin partial gene was used. The reference isolate RBG 1603 Q27 was used as a positive control. The pair of primers VER1-VER2 produced a band of the expected size - 578 bp in 18 isolates, while using FV-F2/FV-R, a 370bp long band confirmed the presence of F. verticillioides in 16 samples. Sixteen out of 18 isolates verified with VER1-VER2 were also identified as F. verticillioides with FV-FS/FV-R. No amplification was observed in a negative control., Tokom perioda od 2014. do 2017. godine sakupljani su uzorci pšenice sa fuzarioznih klasova sa 20 lokacija u Srbiji. Nakon izolacije, gljive su gajene na KDA podlozi 7 dana u termostatu na temperaturi od 25oC. Na osnovu identifikacije in situ na KDA (krompir dekstrozni agar), 36 izolata sekcije Liseola odabrano je za dalje analize. Potvrđena je patogenost svih izolata na listovima pšenice. Virulentnost izolata utvrđena je merenjem dužina formirane pege na mestu inokulacije. Za dokazivanje prisustva vrste Fusarium verticillioides korišćen je par prajmera FV-F2/FV-R koji umnožava sekvence gaoB gena i koji se pokazao kao specifičan za navedenu vrstu. Takođe, sa istim ciljem korišćen je par prajmera VER1-VER2 dizajniran na osnovu dela genske sekvence za kalmodulin. Kao pozitivna kontrola korišćen je referentni izolat RBG 1603 Q27. Par prajmera VER1-VER2 obrazovao je traku očekivane veličine (578bp) kod 18 izolata, dok je pomoću para prajmera FV-F2/FV-R traka dužine 370bp potvrdila prisustvo F. verticillioides kod 16 izolata. Šesnaest od 18 izolata koji su identifikovani VER1-VER2 parom, takođe su identifikovani i FV-FS/FV-R prajmerima. U negativnoj kontroli nije bilo amplifikacije.

Durum wheat is a cereal with high range of vitamins, minerals and other nutritional compounds which are important in human nutrition. Durum wheat selection programs in Serbia are smaller in scope compared to bread wheat, but this does not diminish their importance. Good grain quality is essential for the grinding and food industry. Therefore, it is important that durum wheat is safe to consume. Fumonisins are mycotoxins synthesized mainly by fungi of the genus Fusarium. These toxins occur usually in maize and maize products, although their presence has also been reported in other cereals such as wheat and barley. In a five years period, 2015-2019, durum wheat samples were tested for the presence of fumonisin. Samples were collected from different plots in the Zemun Polje. Each year, 10 randomly selected durum wheat grain samples were tested. The grains were dried, ground and analyzed for the presence of fumonisin. The analysis was performed by ELISA (Enzyme-linked immunosorbent assay) using a kit for fumonisin, Tecna, Italy. The highest concentrations of fumonisin were recorded in 2016 when the values ranged 30.602-43.930 ppm. Medium concentrations (8.872-18.477 ppm) of synthesized fumonisin were recorded in 2015, while in other years (2017, 2018 and 2019) fumonisins were observed in low concentrations (0.024-6.124 ppm). Since previous analyzes have shown that fumonisins are present in durum wheat every year, it is necessary to continue monitoring their appearance in durum wheat, as well as in other small grains.

During the past seven decades, the Maize Research Institute, Zemun Polje has been succeeding in keeping up with global trends within the fields of developing and producing hybrids of various FAO maturity groups (100-700). Considering the length of the growing season, breeding of such a wide range of hybrids, has provided marketing of hybrid maize seed all over the world. Mercantile corn production in Serbia increasingly involves the participation of medium early hybrids FAO 300-400. These hybrids should be characterized as high and stable yields with a quick release of moisture from the kernels. The intensification of agricultural production, higher inputs, harvesting corn directly by shelling in grain, high temperatures accompanied by dry winds and lack of rainfall during the growing period are among the most important causes for the growing medium early hybrids. Medium early hybrids have a smaller plant which enables them growing in higher densities that is, larger number of plants per unit area. Also, due to the shorter growing season, these hybrids go through a period of grain filling in terms of better soil moisture supply. In the Maize Research Institute, Zemun Polje in recent times, the development of medium early maturity hybrids has been performed, and the hybrid ZP 4242 (FAO 400) is a result of such development. This hybrid was tested in trails of the Commission for the Variety Releasing in 2020 and 2021. During the investigation period, yields of the hybrid ZP 4242 were significantly higher than yields of check hybrids NK PAKO and ZEROS. ZP 4242 had a moisture level content on the level of check hybrids. Because of its adaptability, yield stability and good level of drought tolerance new ZP hybrid has good potential for production in different agro-ecological environments.

In recent years, public attention has increasingly focused on the production and consumption of high quality safe food. Changes in the dietary trend have influenced the formation of specific market requirements that have led to the fact that in the diet are increasingly used alternative cereals of high nutritional value, in addition to conventional. The ancient wheat species spelt [Triticum aestivum subsp. spelta (L.) Thell.] has a growing interest due to its various health benefits. Due to its biological and agronomic characteristics, spelt takes an important place among alternative cereals. The aim of this study was to investigate the natural occurrence of pathogenic and toxigenic fungi on seven breeding lines of spelt grains in 2021 grown in Zemun Polje, Serbia. Based on morphological properties (colony and spore appearance) it was determined that breeding line 6337 was the least infected (2,2%) and that the peeled grains had a lower degree of infection compared to grains with glumes. Glumes have been shown to be a physical protection against pathogens. Mycological analyses confirmed the presence Aspergillus spp., Alternaria spp., Fusarium spp. which were the most prevalent. Considering the average values, the most frequent were Aspergillus section Nigri (1,64%). Significance of Aspergillus section Nigri is reflected not only in the deterioration of spelt grain yield, quality and large economic losses but also in the fact that many species of this genus produce toxic metabolites (mycotoxins), which are harmful to human and animal health. Climate change and high adaptability and resistance of toxigenic Aspergillus species are cited as the reason for this phenomenon.

Fusarium infection and deoxynivalenol (DON) contamination on the kernels of seven durum wheat lines (six domestic durum lines ZP 16, ZP 34, ZP 41, ZP 74, ZP 120, ZP DSP 66, and one international durum line Cimmyt 7817) during harvest in two growing seasons (2015- 2016) have estimated. Mycological methods were performed to determine the incidence of Fusarium spp., while the Enzyme-Linked Immunosorbent Assay (ELISA) was used to quantify the total level of DON. Analysis of data was done by statistical method ANOVA (analysis of variance). Tukey’s test was used to compare means at a significance level of 5%. Correlation analyses were performed by Pearson’s test. Based on morphological characteristics, four Fusarium species, F. graminearum, F. proliferatum, F. sporotrichioides, and F. verticillioides, were identified in 2015. A different structure of the Fusarium population, which in addition to F. graminearum, F. sporotrichioides and F. verticillioides, also consisted of F. poae, F. semitectum, and F. subglutinans, was identified in 2016. F. graminearum was the predominant species and the most common cause of Fusarium head blight (FHB) and the primary producer of DON. Other Fusarium spp. Were isolated sporadically and in a low incidence in the kernels. Fungal species from the genera Alternaria, Aspergillus, Chaetomium, Epicoccum, Nigrospora, and Penicillium have also been isolated. The incidence of F. graminearum and level of DON were significantly affected by the wheat genotypes and investigated years. However, obtained results for these parameters were negatively correlated. Thus, in 2015, the incidence of F. graminearum was significantly higher (75.86%) than in 2016 (63.43%), while the level of DON was significantly higher in 2016 (3.636 mg/kg) compared to 2015 (1.126 mg/kg). The highest and the lowest incidence of F. graminearum was on the kernels of line ZP DSP 66 (73%) and line ZP 34 (64.50%), respectively. The highest DON level was 3.854 mg/kg (line ZP 120), and t

The aim of the present study was to determine the presence of toxigenic fungi and mycotoxins in the durum wheat spikes. Their occurrence was observed in the location of Zemun Polje during two production years (2018 and 2019). Samples of two different durum wheat varieties were drawn from several plots and surface-sterilised (solution of diluted bleach and water in the ratio 1:3). Kernels were placed on potato dextrose agar (PDA) in Petri dishes. One hundred kernels were taken from each sample (10 dishes with 10 kernels). After the 7-day growth in the thermostat at the temperature of 25oC, the identification of pathogens was initiated. The following genera of fungi were established in 2018 samples: Alternaria (52.7% of the total number of kernels), Fusarium (34.16%), Aspergillus (5.88%), Nigrospora (4.5%), Acremoniella (1.25%), and others (1.51%). During 2019, the highest percentage (62.6%) of identified fungi belonged to the genus Fusarium, and then to genera Alternaria (29.3 %), Epicoccum (2.8%), Aspergillus (1.7%), and others (3.6%). The drawn samples were used to perform the analysis of mycotoxins for the presence of fumonisins and aflatoxins. The ELISA was applied in this analysis. The results obtained from 2018 samples showed low values for fumonisin. The values of synthesised fumonisin ranged from 0.214 to 1.316 ppm, but the values of synthesised aflatoxin were much higher and ranged from 6.052 to 15.728 ppb. The values of synthesised fumonisin were little higher in 2019 (1.066 - 4.652 ppm), but the values of aflatoxin were lower than year before (0-2.334 ppb).

Fungi of the Fusarium genus belonging to the Liseola section can synthesise fumonisins of greater or smaller concentrations. Maize is a primary host of these species. In recent years their presence has been also observed in small grains. The aim of this study was to observe the concentrations of fumonisins synthesised by small grains after artificial inoculation. Twelve isolates of the following species were selected from the collection of fungal cultures of the Maize Research Institute, Zemun Polje for artificial inoculation: Fusarium verticillioides (6), Fusarium subglutinans (3) and Fusarium proliferatum (3). The concentrations of synthesised fumonisins were analysed in four small grains: wheat (Aurelia), barely (Nektar), triticale (Zenit) and durum (Cosmostar). The artificial inoculation was performed with the hand sprayer when more than a half of tested plants were in the full-blossom stage. Inoculation of plants was done in four replications. The amount of inoculum (spore concentration was 1x10-6 per 1 ml) was 20 ml per a group of 20 spikes. The isolate of Fusarium graminearum species was used for spike inoculation in the positive control, while sterile distilled water was used in the negative control. Inoculated spikes were covered with wet PVC bags that were removed after 48h. After harvest, fumonisins were analysed by the ELISA test (Tecna, Italy). According to obtained results not a single isolate of the observed species synthesised fumonisins in the barley crop. In the remaining crops, isolates of F. subglutinans species synthesised fumonisins in low concentrations (0.793-24.949 ppm), while the corresponding values of isolates of F. proliferatum species were high and ranged from 35.886 to 60.000 ppm. Isolates of F. verticillioides species had low values in wheat (2.162-7.925 ppm), while these values in durum were high (29.610-47.174 ppm). The mean values of synthesised fumonisins in the triticale crop were low (1.357-32.587 ppm).

The objective of the present study was to investigate the susceptibility of maize hybrids to the natural mycotoxins contamination: aflatoxin total (AFLA), deoxynivalenol (DON), zearalenon (ZEA) and fumonisins (FB). Hybrids were grown during two production years (2019 and 2020) in two locations. Mycotoxin contamination of maize grains was evaluated in five sweet maize hybrids. Contamination level of investigated hybrids of first cluster was mainly by below average values of ZEA (all equal to zero) and DON. Hybrids PK4 (S) 2020, PK6 (MS) 2020 and PK4 (MS) 2020 had below average values for AFLA, while remaining treatments of this cluster have mostly elevated values of this mycotoxin. Contamination level of investigated hybrids of second cluster mostly had increased values of mycotoxins ZEA and DON and below average values of AFLA. Samples did not contain fumonisins. Mycotoxin contamination were significantly affected by hybrids and years. We have established that DON and ZEA levels were influenced by the environmental conditions. There were no significant effects of location on the level of AFLA in the sweet maize hybrids. The variation in the properties of mycotoxin content (DON) was significantly influenced by hybrids, and there was no significance of hybrids x location interaction. Differences were more expressed for the content of ZEA and AFLA compared to the content of DON. Hybrid PK1 had the lowest content of DON, while it had the highest content of ZEA. Mycotoxin analyses showed that in all tested hybrids, levels of AFLA, DON, ZEA and FBs were below the maximum permissible levels stipulated by the legislation of the European Union and the Republic of Serbia in maize intended for direct human consumption. These results confirmed that the susceptibility of hybrids is one of the important risks, in addition to climatic factors, for the appearance of toxigenic fungi and their mycotoxins. Genotype tolerance is very important as a preventive measure, which indicates t, Cilj ovog rada bio je da se prouči osteljivost hibrida kukuruza na prirodnu kontaminaciju mikotoksinima (aflatoksin B1 (AFB1), deoksnivalenol (DON), zearalenol (ZEA) i fumonizini (FB)). Hibrdi su gajeni tokom dve proizvodne godine (2019. i 2020) u dve lokacije. Kontaminacija zrna kukuruza mikotoksinima ispitivana je na pet hibrida kukuruza šećerca. Nivo kontaminacije proučavanih hibrida prvog klastera bio je uglavnom ispod prosečnih vrednosti ZEA (sve vrednosti su bile nula) i DON. Vrednosti AFLA kod hibrida PK4 (S) 2020, PK6 (MS) 2020 i PK4 (MS) 2020 su bile ispod prosečne vrednosti, dok su vrednosti ovog mikotoksina bile više od prosečnih vrednosti za ostale tretmane ovog klastera. Nivoi kontaminacije proučavanih hibrida drugog klastera su uglavnom bili viši za mikotoksine ZEA i DON i niži za AFLA. Fumnonizni nisu utrvđreni u uzorcima. Na kontaminaciju mikotoksinima značajno su uticali hibiridi i godine. Utvrđeno je da su uslovi sredine uticali na nivoe DON i ZEA. Lokacija nije značajno uticala na nivo AFLA kod hibrida kukuruza šećerca. Hibridi su značajno uticali na variranje sadržaja mikotoksina (DON), dok hibrid × lokacija interakcija nije bila značajna. Razlike su bile izraženije za sadržaj ZEA i ALFA nego za sadržaj DON. Najniži sadržaj DON utvrđen je kod hibrida PK1, kod koga je sadržaj ZEA bio najviši. Analize mikotoksina pokazuju da su nivoi AFLA, DON, ZEA i FB u svim ispitivanim hibridima bili ispod maksimalno dozvoljenih nivoa koji su propisani zankonima Evropske Unije i Republike Srbije za kukuruz koji je namenjen za direktnu ljudsku konzumaciju. Ovi rezultati potvrđuju da je osetljivost hibrida jedan od važnih rizika pored klimatskih faktora za pojavu toksigenih gljiva i njihovih mikotoksima. Tolerantnost genotipa je veoma važna preventivna mera, na koju oplemenjivači moraju da obrate pažnju u programima oplemenjivanja kukuruza šećerca.

In Serbia, aspergillus ear rot caused by the disease pathogen Aspergillus parasiticus (A. parasiticus) was first detected in 2012 under both field and storage conditions. Global climate shifts, primarily warming, favour the contamination of maize with aflatoxins in temperate climates, including Serbia. A five-year study (2012–2016) comprising of 46 A. parasiticus strains isolated from maize kernels was performed to observe the morphological, molecular, pathogenic, and toxigenic traits of this pathogen. The HPLC method was applied to evaluate mycotoxin concentrations in this causal agent. The A. parasiticus isolates synthesised mainly aflatoxin AFB1 (84.78%). The percentage of isolates synthesising aflatoxin AFG1 (15.22%) was considerably lower. Furthermore, the concen- tration of AFG1 was higher than that of AFB1 in eight isolates. The polyphase approach, used to characterise isolates, showed that they were A. parasiticus species. This identification was verified by the multiplex RLFP-PCR detection method with the use of restriction enzymes. These results form an excellent baseline for further studies with the aim of application in the production, processing, and storage of cereal grains and seeds, and in technological processes to ensure the safe production of food and feed.

The application of pesticides is an actual plant protection measure in agriculture that can have adverse effects on people’s health and environments. A great attention is paid to the biological fungicides. Performed studies indicate satisfactory results of activities of essential oils, which furthermore point out to possibilities of their inclusion in crop protection programs. The aim of this study was to determine antifungal, contact activity of essential oils of thyme (Thymus vulgaris L.) and oregano (Origanum vulgare L.) on toxigenic fungal species Aspergillus flavus causing maize kernel rot. The sterile filter paper was placed in the inner lead of Petri dish. Two, four, six, eight and 10 µl of essential oils were pipetted on the paper, and then pure A. flavus cultures were subcultured on PDA. After seven days in the dark at 25°C, the degree of inhibition was determined by measuring the fungal growth and their comparison to the control. The fungus, without addition of essential oils, was used as the control. The strongest antifungal activity was expressed by thyme essential oil, which already at the amount of 2 µl completely inhibited the mycelium growth. The identical effect was achieved with the amount of 4 µl. The greater amount of essential oil the more progressive growth of the fungal colony (6 µl – 3 mm; 8 µl – 9 mm). However, the mycelium growth at the amount of 10 µl of essential oil was only 1 mm. The similar results were gained with oregano essential oil (2 µl – 3 mm; 4 µl – 7 mm; 6 µl – 9 mm; 8 µl – 9 mm; 10 – 6 mm). The fungal growth of control was 30 mm. The obtained results indicate the significant potential of the application of thyme and oregano essential oils as possible natural and environmentally friendly means for the protection of maize against A. flavus.

Barley (Hordeum vulgare L.) is a secondary grain crop in Serbia used commercially for animal feed, seed, and human food applications. The production of barley in the 2016 to 2017 growing season reached a record yield of almost 400,000 metric tons (USDA 2017). Aspergillus contamination has been rare in the agroecological conditions of cereal-growing areas in Serbia. Changes in climatic factors, such as occurrence of high temperatures and prolonged droughts, increased frequency of Aspergillus spp. Species Aspergillus parasiticus was isolated from maize grain for the first time in Serbia in 2012 and from wheat grains in 2017 (Nikolic et al. 2018). We hypothesized that these pathogens can also be present in barley fields in Serbia. Barley spikes exhibiting bleaching were sampled at the beginning of June 2017 grown in northern Serbia. In severe infections, barley spikes get a dry look with awns that stand upright and firm. The incidence of the disease of the bleached spiked in the field was 15 to 20%. From each representative sample, 100 shriveled grains were collected. After surface sterilization with bleach/distilled water 1:3, 100 grains per sample (10 per Petri dish) were placed on potato dextrose agar and incubated at 25°C for 7 days. After isolation, 50% of isolates were identified as Alternaria spp., 20% as Fusarium spp., approximately 15% as Aspergillus spp. In order to reliably identify individual species of fungi, the fragments of colonies were transferred to malt extract agar (MEA) and Czapek yeast agar (CYA) and incubated in the dark for 7 days. The fungal colonies were dark green. The reverse side was pale yellow. The average colony diameter was 65 mm. Conidia were spherical and rough with thick walls. The average size of conidia was 5.1 µm. Colonies were floccose and thin on MEA and CYA. Based on growth and morphological characteristics, isolates were determined as A. parasiticus (Pitt and Hocking 2009). Molecular detection of Aspergillus species was done b

Animal welfare continues to be an important issue in the whole world. Therefore the production of safe feed remains one of the most important tasks because the consumption of contaminated feeds by livestock has been associated with a variety of adverse health effects including feed refusal, reduced weight gain, diarrhoea and emesis. Wheat bran, a by-product of the dry milling of common wheat (Triticum aestivum L.) into flour, is one of the major agro-industrial by-products used in animal feeding. In addition, wheat bran can also been used in human diet as a source of fiber in the preparation of high-fiber pasta, noodles etc. Deoxynivalenol (DON) is one of several mycotoxins produced by certain Fusarium species that frequently infect wheat, corn, rice, oats, barley and other grains in the field or during storage. In this study, a total of 72 samples of wheat bran were collected from several producers from Serbia in the period 2019-2020. The samples were analysed for deoxynivalenol (DON) content by using an enzyme-linked immunosorbent assay Ridascreen® Fast DON SC. The occurrence of DON in the tested samples was 100%, with average concentration 2.79 mg kg-1 and maximum DON content of 9.12 mg kg-1 . These results suggest very high level of DON concentration in all tested samples. The limit of 8.00 mg kg-1 imposed by Serbian legislation for DON content in feed was exceeded in two studied samples. However, only fourteen of them (19%) would be suitable for human consumption, due to a much lower allowed limit (0.75 mg kg-1 ), which raises a risk for consumers.

Initial studies related to cytoplasmic male sterility (CMS) were performed by Rhoeds in 1931. CMS is used in maize to enhance efficiency of seed production with simultaneous cost redaction. The majority of studies showed positive effects of CMS on maize grain yield. Grain yields recorded male sterile plants were higher by 5-10% than the ones in female fertile plants. Seven maize inbred lines of different origin and growing season were analysed. Each inbred was analysed in five variants: original inbred (N), CMS-C, RfC, CMS-S and RfS. The aim of this study was to compare grain yields and yield components of original inbreds and their CMS and Rf variants. The highest yields of observed inbreds (5.303 and 5.197 t/ha) were recorded in those with C and S cytoplasm, respectively. According to the LSD test, at significance levels of 0.05 and 0.01, the longest ears of 15.57, 15.56 and 15.46 cm were detected in inbreds with original, S and C cytoplasm, respectively. The highest kernel row number (12.98) at both significance levels was established in inbreds with C cytoplasm. The highest number of kernels per row (33.92) at both levels of significance was recorded in inbreds with S cytoplasm. The greatest kernel depth (0.8212 and 0.8196 cm) at both significance levels was established in inbreds with C and S cytoplasm, respectively. The greatest 1000-kernel weight at both levels of significance was detected in inbreds with normal cytoplasm. The highest number of kernels per m2 (2716 and 2676) was recorded in inbreds with C and S cytoplasm, respectively.