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The goal of this work was to characterize potential biological agents responsible for biodeterioration in heritage buildings identified as Costa Rican Caribbean architecture using a multidisciplinary approach. First, an inventory of buildings of Caribbean architecture from Limón City was elaborated. Then, a multi-criteria evaluation system was used to define priority scores to select buildings for sampling and analysis. Selected buildings were sampled for potential biological agents responsible for biodeterioration. Wood samples were taken from sections showing visual damage and fungi were isolated and characterized using Sanger sequencing. A basic profile of Costa Rican Caribbean architecture was established. A preliminary identification of 101 buildings that satisfied the basic profile of characteristics was con-ducted. Subsequently, these buildings were classified into 4 typologies from which 23 buildings belonging to Typologies 1 and 2 were chosen for the collection and registration of data. Applying the multi-criteria evaluation, 6 buildings with final scores higher than 60 points were selected from which the 1st (Casa Misionera Bautista) and 3rd (Antigua Capitanía de Puerto) in the classification were selected for further characterization of biological agents. From the sampling conducted on these two buildings. A total of 36 fungi were isolated. All fungi identified belonged to the fungal group Ascomycota phylum, the most common genera being Trichoderma, Fusarium, and Penicillium. The study shows how multidisciplinary studies can improve the process of decision-making in the preservation of heritage with the use of a ranking obtained for each building in the multi-criteria evaluation and the use of advanced techniques for the identification of biological agents.

Social Impact Statement Vanilla planifolia is the source of the spice vanilla. This study is part of an international initiative to study the biology, including mycorrhizal fungi and cultivation practices of vanilla to improve its production in Mexico. The study focused on documenting mycorrhizal fungal diversity in vanilla. It also provided preliminary data on differences in mycorrhizal fungal communities associated with cultivation practices. A richer mycorrhizal community was observed in vanilla growing in a wild natural farm compared with those from a highly managed farm. Our research provides insights for sustainable vanilla production that can benefit Mexican farming communities. Summary Relatively little is known regarding differences in root symbionts (i.e., mycorrhizae) between epiphytic and terrestrial orchids. We characterized the mycorrhizal fungal communities of aerial and terrestrial roots of the orchid, Vanilla planifolia, from four Mexican farms representing different management systems. Amplicon sequencing identified 40 putative mycorrhizal fungi based on ITS sequence data, these included traditional orchid mycorrhizal fungi such as Ceratobasidium, and Thanatephorus in the order Cantharellales as well as Serendipitaceae in the order Sebacinales, and species of several genera traditionally considered as ectomycorrhizal fungi. Mycorrhizal fungal communities were similar in aerial and terrestrial roots, but differed in read abundances. Plants growing in wild‐natural conditions hosted a richer, but not statistically different, community of mycorrhizal fungi in comparison with plants in the highly managed farm. Soil characteristics including texture, organic matter, N, P, and K do not explain differences between fungal communities at these farms. This is one of the first reports of a diverse community of fungi traditionally considered to form ectomycorrhizas in association with aerial orchid roots. Further research is needed to understand the functional role of these putative mycorrhizal fungi in the ecology of V. planifolia, and if ectomycorrhizal fungi commonly occur in other hemiepiphytic and epiphytic orchids.

Abstract Background The dominant fungi in arid grasslands and shrublands are members of the Ascomycota phylum. Ascomycota fungi are important drivers in carbon and nitrogen cycling in arid ecosystems. These fungi play roles in soil stability, plant biomass decomposition, and endophytic interactions with plants. They may also form symbiotic associations with biocrust components or be latent saprotrophs or pathogens that live on plant tissues. However, their functional potential in arid soils, where organic matter, nutrients and water are very low or only periodically available, is poorly characterized. Results Five Ascomycota fungi were isolated from different soil crust microhabitats and rhizosphere soils around the native bunchgrass Pleuraphis jamesii in an arid grassland near Moab, UT, USA. Putative genera were Coniochaeta, isolated from lichen biocrust, Embellisia from cyanobacteria biocrust, Chaetomium from below lichen biocrust, Phoma from a moss microhabitat, and Aspergillus from the soil. The fungi were grown in replicate cultures on different carbon sources (chitin, native bunchgrass or pine wood) relevant to plant biomass and soil carbon sources. Secretomes produced by the fungi on each substrate were characterized. Results demonstrate that these fungi likely interact with primary producers (biocrust or plants) by secreting a wide range of proteins that facilitate symbiotic associations. Each of the fungal isolates secreted enzymes that degrade plant biomass, small secreted effector proteins, and proteins involved in either beneficial plant interactions or virulence. Aspergillus and Phoma expressed more plant biomass degrading enzymes when grown in grass- and pine-containing cultures than in chitin. Coniochaeta and Embellisia expressed similar numbers of these enzymes under all conditions, while Chaetomium secreted more of these enzymes in grass-containing cultures. Conclusions This study of Ascomycota genomes and secretomes provides important insights about the lifestyles and the roles that Ascomycota fungi likely play in arid grassland, ecosystems. However, the exact nature of those interactions, whether any or all of the isolates are true endophytes, latent saprotrophs or opportunistic phytopathogens, will be the topic of future studies.

White-nose syndrome (WNS) is a bat disease caused by the fungal pathogen Pseudogymnoascus destructans, which thrives in cold and very humid environments where bats frequently hibernate. Conidia of Pseudogymnoascus species are often documented on bats prior to the onset of WNS, but characterization of high-risk areas defined by microclimate cave conditions have been lacking. Investigating the occurrence of this fungal genus and appropriate environmental conditions to support P. destructans in southwestern U.S. caves is key to understanding the sites most likely to be impacted by WNS. Microclimate conditions in ten caves at El Malpais (ELMA) National Monument in New Mexico, USA were recorded using i-Button data loggers during the winters of 2011–2014 to assess appropriate environmental conditions (temperature and relative humidity) for P. destructans and other Pseudogymnoascus species. Optimal microclimate conditions for P. destructans and other psychrophilic fungi were found in all the caves with at least 50% of the caves identified as high-risk areas. Pseudogymnoascus species were detected in 70% of the caves using culturing methods and PCR, but no soil samples were positive for P. destructans using real-time PCR in soil and guano samples. Pseudogymnoascus destructans has a recognized range of appropriate temperatures and relative humidity for growth and cave microclimate can help define high-risk areas. This study offers resource managers guidance for establishing priority monitoring areas in their bat caves to determine which bat species are at higher risk.

Microorganisms that reside on and in mammals, such as bats, have the potential to influence their host’s health and to provide defenses against invading pathogens. However, we have little understanding of the skin and fur bacterial microbiota on bats, or factors that influence the structure of these communities. The southwestern United States offers excellent sites for the study of external bat bacterial microbiota due to the diversity of bat species, the variety of abiotic and biotic factors that may govern bat bacterial microbiota communities, and the lack of the newly emergent fungal disease in bats, white-nose syndrome (WNS), in the southwest. To test these variables, we used 16S rRNA gene 454 pyrosequencing from swabs of external skin and fur surfaces from 163 bats from 13 species sampled from southeastern New Mexico to northwestern Arizona. Community similarity patterns, random forest models, and generalized linear mixed-effects models show that factors such as location (e.g., cave-caught versus surface-netted) and ecoregion are major contributors to the structure of bacterial communities on bats. Bats caught in caves had a distinct microbial community compared to those that were netted on the surface. Our results provide a first insight into the distribution of skin and fur bat bacteria in the WNS-free environment of New Mexico and Arizona. More importantly, it provides a baseline of bat external microbiota that can be explored for potential natural defenses against pathogens.