Epiphytic microflora and mycotoxin content in meadows–Is plant biodiversity affecting fungal contamination?

mindset to focus not only on yield of forage production, but on ecosystem services as well [2]. The biggest issue with the wider use of flowering meadows for feeding delves in the insufficient information about quality of such feed. Quality of the produced feed depends mainly on the quality of inputs, e.g. forage grasses and legumes. Fodder quality has a decreasing tendency each month throughout the year with lowest quality in the oldest fodder stands. Study of Kononenko and Burkin [4] shows, that the highest values of secondary metabolites, which are a common cause of health complications and toxicosis in cattle, occur more frequently in the autumn. Additionally, it is possible to see the emergence of mycotoxins in the areas where they were not detected in the past as a result of climate change [5–7]. Levels of mycotoxin contamination of forage stands changes depending on the dominant species in the mixture. Study of Kononenko and Burkin [8] states, that mycotoxins were pres- ent in all mixtures. Furthermore, they were detected in at least 50% of the samples. In the mix- tures where legume Vicia sp. was dominant was deoxynivalenol (DON) absent. Another study adds that agroforestry can change the abundance of soil bacteria and fungi in comparison with a monoculture and open grassland [9]. This suggests that number of grown species may affect epiphytic microbiota as well. Mycotoxicosis is often difficult to diagnose due to its non-specific symptoms and symp- toms’ similarity with other clinical diagnoses. Therefore, the mycotoxins’ effects on animal

health can often escalate to death of an animal due to a slow diagnosis and often incorrect treatment. This goes hand in hand with the welfare aspect of animal husbandry. Subsequently it is important to note the economic impact on production by organ damage and decrease in weight gain, feed efficiency and reproduction rate [10]. Clinical symptoms such as anorexia, impaired rumen functionality, ankle adduction, posterior paralysis, dairy cow syndrome etc. often occur [11]. Research interest in mycotoxin presence in fresh forage is limited in comparison with mycotoxin contamination of cereals [12]. In the light of current information, it is important to evaluate the quality of species-rich mixtures in comparison to monocultures for increasing the their demand and growing area. We postulated that in diverse ecosystem, such as species-rich mixtures, will be microorganisms, and especially fungi, more abundant. This, moreover, may lead to higher levels of mycotoxin contamination in comparison with monocultures. Material and methods The experiment was established in 2018 on Mendel University in Brno’s Research station Vatı´n, located in in the Bohemian-Moravian Highlands (49˚52’ N, 15˚96’ E). Site is located 560 m above sea level. Average yearly precipitation is 617 mm and average yearly temperature is 6.9˚C. Monthly weather conditions can be found in Fig 1. Soil class on the site is loamy- sand, soil type is cambisol on the diluvium of piotic orthogenesis. Small plots used in the experiment had 10 m2 (1,25x8 m), no fertilization was applied in any of the variants. Each vari-

ant was established in triplets in Latin square design. Sampling was done 8. 10. 2020. Following types of 2-year old stands were studied: • Grass monocultures: Festuca arundinacea Schreb. (in the varieties ’Prosteva’, ’Kora’, ’Apri- lia’, ’Philia’) and Lolium perenne L. (in the variety ’Promed’) • Species-rich meadow mixtures comprising up to 27 plant species (Table 1): ⚬ Mixture 1: 0% grasses + 40% legumes + 60% forbs ⚬ Mixture 2: 70% grasses + 10% legumes + 20% forbs PLOS ONE Biodiversity and plant fungal contamination PLOS ONE | https://doi.org/10.1371/journal.pone.0288397 September 14, 2023 2 / 11