Maize (Zea mays L.) is one of the most important agricultural species in the world, providing a staple food and is used as a source of income for many populations in developing countries [1]. Despite the clear advantages of improved varieties, the performance of a maize genotype and the potential of maize yield is largely determined by a specific combination of different factors, and is affected by abiotic and biotic stresses, such as climatic factors, pests, soil characteristics, solar radiation, field management practices, and the seed quality and genetic potential of the hybrid [2]. However, the impact of climate change in agricultural production is undoubtable. Variations in climatic conditions frequently favor the multiplication of pathogens while negatively affecting soil fertility and plant productivity. Climate change is bringing new species of diseases and pests that do not have any control methods fully developed yet [3]. The variability of climatic conditions contributes to higher biosynthesis of mycotoxins in maize, causing economic losses in production and risk for human and animal health [4]. Numerous species belonging to the genus Aspergillus are widely distributed worldwide, both in soil, as well as in various agricultural crops, especially maize and plant products [5]. Maize susceptibility to Aspergillus ear rot and aflatoxin accumulation presents a global economic and health problem. The most important species that causes Aspergillus ear rot is Aspergillus flavus. Although the species A. flavus is a saprophyte, under favorable conditions for development it can cause significant rotting of corn ears and kernels in the field, as well as during storage. The species A. flavus has the ability to synthesize aflatoxins, which are classified as the most toxic natural substances [6]. Food contaminated with aflatoxins poses a serious risk to human and animal health. Aflatoxin B1 is the strongest known carcinogen and is classified as a group 1 carcinogen by the International Agency for Research on Cancer [7]. Consumption of food contaminated with aflatoxin is one of the main causes of liver cancer in the world [8,9]. In addition to the carcinogenic effect, aflatoxins exhibit strong mutagenic, teratogenic, and immunosuppressive effects [10]. A. flavus is a xerophilic fungal species that has developed physiological mechanisms for adaptation in stressful environmental conditions [11]. High average temperatures and long dry periods lead to heat stress in plants and increased aflatoxin contamination [12,13]. Another important factor that contributes to intense appearance of A. flavus on corn grain are insects that represent the vector of conidia transmission, but also mechanically damage the grain, which facilitates the penetration of the pathogen into unprotected endosperm of the grain [5,14]. In order to reduce the aflatoxin occurrence in the field, preventive measures which are recommended include the selection of a suitable maize hybrid with increased tolerance to abiotic and biotic stress, timely sowing, crop rotation, proper plant nutrition, irrigation, control of insects, diseases and weeds [15,16]. The most effective measure to reduce Aspergillus ear rot and aflatoxin contamination is the cultivation of resistant maize hybrids. Therefore, numerous research studies are focused on the discovery of new sources of resistance [17,18,19,20,21,22]. There are various approaches to develop aflatoxin resistant hybrids, including molecular techniques [23]; antifungal proteins studies [24]; and studies of the morphological characteristics of ear and grain in resistant hybrids [25]. Since A. flavus infection is associated with drought stress, one approach is the development of drought-resistant hybrids [26,27]. Several studies have focused on the effectiveness of using insect-resistant hybrids to indirectly reduce aflatoxin accumulation [28,29]. The main difficulty in developing resistant maize hybrids to A. flavus is the strong interaction between genotype and environment [30]. Therefore, it is necessary to identify maize genotypes that possess stable resistance in a wide range of environmental conditions [31]. The improvement of artificial inoculation techniques could represent one of the most promising methods for the successful identification of resistant maize genotypes to Aspergillus ear rot and aflatoxin production. Therefore, the aim of this research was to evaluate the sensitivity of different maize hybrids to A. flavus infection and subsequent aflatoxin accumulation.