Ant's Nest as a microenvironment: Distinct <i>Mucoromycota</i> (Fungi) community of the red wood ants' (<i>Formica polyctena</i>) mounds

Many social insect species build nests, which differ from the surrounding environment and are often occupied by specific organismal communities. These organisms may interact mutualistically or parasitically with the nest-builders, or simply co-occur, being able to survive in these microenvironments. In temperate forests, red wood ants (e.g. <i>Formica polyctena</i>) are known to create distinct, highly developed nests, which consist of large, above-ground mounds, built primarily out of plant matter collected from the forest litter. The microorganismal communities of such mounds remain understudied. As representatives of <i>Mucoromycota</i> fungi commonly engage in the decomposition process of the forest litter, they would be expected to occur in the mounds. However, it is still not known whether the <i>Mucoromycota</i> community of these ants' nests differ from the one of the surrounding forest litter. In order to distinguish mound-associated taxa, we characterized <i>Mucoromycota</i> communities of <i>Formica polyctena</i> mounds and the surrounding forest litter. We sampled four sites, twice in a season. Sampled material was plated on agar media and emerging <i>Mucoromycota</i> colonies were identified based on their morphology. Fungal identification was further confirmed using DNA barcoding. In order to compare described communities, PERMANOVA test and non-metric multidimensional scaling ordinations were used. To distinguish taxa associated with the mounds, multilevel pattern analysis was performed. Our results show that the <i>Mucoromycota</i> community of <i>Formica polyctena'</i>s mound differs from the community of the surrounding forest litter. While representatives of <i>Entomortierella lignicola</i> and <i>Absidia cylindrospora</i> clade were found to be associated with the mound environment, representatives of <i>Umbelopsis curvata</i> and <i>Podila verticillata-humilis</i> clade were associated with forest litter, and were rarely present in the mounds. Our findings strongly suggest that the red wood ants' nest is a specific microenvironment in the temperate forest floor, which is a preferred microhabitat for the mound-associated <i>Mucoromycota</i>, possibly adapted to live in proximity to ants.