Intensification of livestock farming creates conditions for the accumulation and circulation of pathogenic and opportunistic fungi with the development of antifungal resistance. The study aimed to perform comprehensive assessment of fungal contamination of production environment in various areas of livestock farms and determine sensitivity of the extracted isolates to common antifungal agent. The presence of fungal contamination in the air of all working areas of the studied production facility was established. Predominance of Aspergillus niger and Candida albicans was revealed. In 50% of cases, mixed contamination with different types of mold fungi was observed. The highest contamination levels were reported for the bedding material. Resistance of microorganisms to several antifungal drugs with different mechanisms of action at once has been revealed. The study confirmed systemic contamination of the livestock complex production environment with potentially pathogenic fungi forming stable communities in feed and bedding. The reported sanitary and microbiological approaches to assessing fungal contamination at livestock production facilities have a pronounced comprehensive preventive focus, which makes it possible to improve the farm employees’ working conditions, as well as provide measures to improve the livestock complex production environment.