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

Phenotype as well as genotype affect the phyllosphere composition [26, 27]. Burkin et al. [4] stated that the amount of mycotoxin in a monoculture depends on the monoculture species. Pieczul et al. [28] studied mycoflora on Arhenantherum aristatum; this species, related to A. odoratum present in our experimental mixture, is infected by many fungi pathogen species, of which Fusarium spp. Is the most important. Presence of this species could therefore increase the risk of high OF counts and subsequent mycotoxin contamination in Mixture 3. Out of 26 identified species found on Polish meadow was the highest level of fungal contamination found in tall fescue and perennial ryegrass, which suggests potential feed safety risk of these meadow species [3]. However, results of our experiment did not find microbial differences between the grass species’ amounts of fungi and yeasts. This is in accordance with Baholet et al. [23], where no significant differences were observed between ryegrass, fescue and timo- thy. On the contrary, TMC in our experiment was significantly higher in ryegrass in compari- son to fescues, which can suggest effects of other factors on the epiphytic microbial communities. Results of Seguin et al. [29] showed, that both fungal species and fungal counts in the sample were highly dependent on the botanical species sampled. Higher fungal counts were observed more often in monoculture hay in comparison with multi-species hay, however, these results depend on the type of multi-species hay observed. This was supported especially

in case of significantly lower fungal contamination was found in perennial ryegrass hay in comparison with Alopecurus geniculatus or Crau and Swiss hay. Bacterial communities have also been proven to show species-specific differences in amounts [30]. Epiphytic microbiota is considered an important quality factor in subsequent processing of biomass into conserved feed [30, 31]. Authors therefore imply that besides floristic composition of a sample and geo- graphical location significantly affect hygienic quality of hay. Biomass gathered from meadows can be colonized by multiple species of fungal mycotoxin producers, including Humicola spp., Alternaria spp., Cladosporium spp., Torula spp., Fusar- ium spp. and Mucor spp. [3]. Birdsfoot trefoil (Lotus corniculatus L.) is a host plant of the Table 5. Average mycotoxin contamination depending on botanical species. Deoxynivalenol (μg kg-1) Aflatoxin B1 (μg kg-1) Zearalenone (μg kg-1) Festuca arundinacea ’Prostela’ (n = 3) Mean <LOQ 0.0502 a 0.4862 a SD <LOQ 0.0016 0.4406 Festuca arundinacea ’Aprilia’ (n = 3) Mean <LOQ 0.0520 a 0.3810 a SD <LOQ 0.0024 0.0706 Festuca arundinacea ’Philia’ (n = 3) Mean <LOQ 0.0500 a 0.3620 a SD <LOQ 0.0070 0.0613 Festuca arundinacea ’Kora’ (n = 3) Mean <LOQ 0.0521 a 0.2955 a SD <LOQ 0.0038 0.0137 Lolium perenne ’Promed’ (n = 3) Mean <LOQ 0.0420 a 0.5896 a SD <LOQ 0.0012 0.0271 Mixture I (n = 3) Mean 1.3307 0.0535 a 0.8930 a SD 0.2061 0.0349 0.6261 Mixture II (n = 3) Mean 2.7228 0.0212 a 0.9140 a SD 1.9976 0.0165 0.1759

Mixture III (n = 3) Mean 1.2348 0.0229 a 0.8082 a SD 0.8555 0.0099 0.1832 Results are expressed as a mean and standard deviation (SD). Mycotoxin concentration marked as <LOQ were under the limit of quantification (DON < 0.03 μg kg-1). Mean values are statistically significant at p < 0.05, when values in columns are marked with a different letter in the upper index. https://doi.org/10.1371/journal.pone.0288397.t005 PLOS ONE Biodiversity and plant fungal contamination PLOS ONE | https://doi.org/10.1371/journal.pone.0288397 September 14, 2023 7 / 11