Abstract
Bats are widespread mammals that play key roles in ecosystems as pollinators and insectivores. However, there is a paucity of information about bat-associated microbes, in particular their fungal communities, despite the important role microbes play in host health and overall host function. The emerging fungal disease, white-nose syndrome, presents a potential challenge to the bat microbiome and understanding healthy bat-associated taxa will provide valuable information about potential microbiome-pathogen interactions. To address this knowledge gap, we collected 174 bat fur/skin swabs from 14 species of bats captured in five locations in New Mexico and Arizona and used high-throughput sequencing of the fungal internal transcribed (ITS) region to characterize bat-associated fungal communities. Our results revealed a highly heterogeneous bat mycobiome that was structured by geography and bat species. Furthermore, our data suggest that bat-associated fungal communities are affected by bat foraging, indicating the bat skin microbiota is dynamic on short time scales. Finally, despite the strong effects of site and species, we found widespread and abundant taxa from several taxonomic groups including the genera Alternaria and Metschnikowia that have the potential to be inhibitory towards fungal and bacterial pathogens.
Similar content being viewed by others
Data Availability
Data and code from this study are available on GitHub: https://github.com/pattyjk/bat_mycobiome.
References
Abarenkov K et al (2010) The UNITE Database for molecular identification of fungi – recent updates and future perspectives. New Phytol 186:281–285
Ange-Stark, Meghan et al (2019) White-nose syndrome restructures bat skin microbiomes. BioRxiv. https://doi.org/10.1101/614842 (April 4, 2023)
Avena CV, Parfrey LW, Leff JW, Archer HM, Frick WF, Langwig KE, Kilpatrick AM, Powers KE, Foster JT, McKenzie VJ (2016) Deconstructing the bat skin microbiome: influences of the host and the environment. Front Microbiol 7:7153
Banerjee A, Baker ML, Kulcsar K, Misra Plowright R, Mossman K (2020) Novel insights into immune systems of bats. Front Immunol 11:26
Bataille A, Cashins SD, Grogan L et al (2015) Susceptibility of amphibians to chytridiomycosis is associated with MHC class II conformation. Proc R Soc B: Biol Sci 282:20143127
Bogan MA, Cryan PM, Valdez EW, Ellison LE, O’Shea TJ (2003) Western crevice and cavity-roosting bats. pp 69–77 Report U.S Geological Survey Reston VA. http://pubs.er.usgs.gov/publication/70180874
Bowen JL, Kearns PJ, Byrnes JE et al (2017) Lineage overwhelms environmental conditions in determining rhizosphere bacterial community structure in a cosmopolitan invasive plantNature. Communications 8:1–8
Buck JC, Weinstein SB, Young HS (2018) Ecological and evolutionary consequences of parasite avoidance. Trends Ecol Evol 33:619–632
Byrd AL, Belkaid Y, Segre JA (2018) The human skin microbiome. Nat Rev Microbiol 16:143–155
Caporaso JG, Lauber CL, Walters WA et al (2011) Global patterns of 16S rRNA diversity at a depth of millions of sequences per sample. Proc Natl Acad Sci 108:4516–4522
Cheng Y, Fox S, Pemberton Hogg C, Papenfuss AT, Belov K (2015) The Tasmanian devil microbiome–implications for conservation and management. Microbiome 3:76
Choudoir MJ, Barberán A, Menninger HL, Dunn RR, Fierer N (2018) Variation in range size and dispersal capabilities of microbial taxa. Ecology 99:322–334
Cho I, Blaser MJ (2012) The human microbiome: at the interface of health and disease. Nat Rev Genet 13:260–270
Clayton JB, Gomez A, Amato K, Knights D, Travis DA, Blekman R, Knight R, Leigh S, Stumpf R, Wolf T, Glander KE, Cabana F, Johnson TJ (2018) The gut microbiome of nonhuman primates: lessons in ecology and evolution. Am J Primatol 80:e22867
Cole JR, Wang Q, Fish JA et al (2014) Ribosomal Database Project: data and tools for high throughput rRNA analysis. Nucleic Acids Res 42:D633–D642
Cornelison CT, Keel MK, Gabriel KT, Barlament CK, Tucker TA, Pierce GE, Crow SA (2014) A preliminary report on the contact-independent antagonism of Pseudogymnoascusdestructans by Rhodococcus rhodochrous strain DAP96253. BMC Microbiol 141:1–7
Cornelison CT, Gabriel KT, Barlament C, Crow SA (2014) Inhibition of Pseudogymnoascusdestructans growth from conidia and mycelial extension by bacterially produced volatile organic compounds. Mycopathologia 1771–2:1–10
Cryan PM, Meteyer CU, Boyles JG, Blehert DS (2010) Wing pathology of white-nose syndrome in bats suggests life-threatening disruption of physiology. BMC Biol 8:1–8
Cuscó A, Belanger JM, Gershony L, Islas-Trejo A, Levy K, Medrano JF, Sánchez A, Oberbauer AM, Francino O (2017) Individual signatures and environmental factors shape skin microbiota in healthy dogs. Microbiome S:139
Csutak O, Vassu T, Cornea P, Grebenisan I (2007) Genetic characterization of two new Metschnikowia strains with antifungal activity. Rom Biotechnol Lett 122:3175
Dura J et al (2014) Karst evolution of the Garraf Massif (Barcelona, Spain): doline formation, chronology and archaeo-palaeontological archives. J Cave Karst Stud 76:69–87
Edgar RC (2010) Search and clustering orders of magnitude faster than BLAST. Bioinformatics 2619:2460–2461
Edgar RC, Haas BJ, Clemente JC, Quince C, Knight R (2011) UCHIME improves sensitivity and speed of chimera detection. Bioinformatics 2716:2194–2200
Fenster SD et al (2019) Examination of fungal diversity present on mexican free-tailed bats, tadarida brasiliensis mexicana, in Colorado. Southwest Nat 63(4):256–267
Frick WF, Pollock JF, Hicks AC, Langwig KE, Reynolds DS, Turner GG, Butchkoski CM, Kunz TH (2010) An emerging disease causes regional population collapse of a common North American bat species. Science 329:679–682
Grisnik M, Bowers O, Moore AJ, Jones BF, Campbell JR, Walker DM (2020) The cutaneous microbiota of bats has in vitro antifungal activity against the white nose pathogen. FEMS Microbiol Ecol 96:193
Hamm PS, Caimi NA, Northup DE et al (2017) Western bats as a reservoir of novel Streptomyces species with antifungal activity. Appl Environ Microbiol 835:e03057-e3116
Holz Peter H et al (2018) Two subspecies of bent-winged bats (Miniopterus Orianae Bassanii and Oceanensis) in Southern Australia have diverse fungal skin flora but not Pseudogymnoascus destructans ed. Michelle L. Baker. PLOS One 13:0204282
Huffnagle GB, Noverr MC (2013) The emerging world of the fungal microbiome. Trends Microbiol 217:334–341
Iacob S, Iacob DG, Luminos LM (2019) Intestinal microbiota as a host defense mechanism to infectious threats. Front Microbiol 9:3328
Ingala MR, Simmons NB, Perkins SL (2018) Bats are an untapped system for understanding microbiome evolution in mammals. mSphere 3:e00397-00418
Johnson LJAN et al (2013) Psychrophilic and psychrotolerant fungi on bats and the presence of Geomyces spp. on bat wings prior to the arrival of white nose syndrome. Appl Environ Microbiol 79:5465–5471
Johnson LJ, Miller AN, McCleery RA, McClanahan R, Kath JA, Lueschow S, Porras-Alfaro A (2013) Psychrophilic and psychrotolerant fungi on bats and the presence of Geomyces spp. on bat wings prior to the arrival of white nose syndrome. Appl Environ Microbiol 79:5465–5471
Kearns PJ, Fischer S, Fernández-Beaskoetxea S et al (2017) Fight fungi with fungi: antifungal properties of the amphibian mycobiome. Front Microbiol 8:2494
Knief C, Ramette A, Frances L, Alonso-Blanco C, Vorholt JA (2010) Site and plant species are important determinants of the Methylobacterium community composition in the plant phyllosphere. ISME J 46:719–728
Kolodny O et al (2019) Coordinated change at the colony level in fruit bat fur microbiomes through time. Nat Ecol Evol 3:116–124
Kunz TH, Kurta A (1988) Capture methods and holding devices. p 1–29. In Kunz TH ed. Ecological and behavioral methods for the study of bats. Smithsonian Institution Press Washington DC
Kueneman JG, Bletz MC, McKenzie VJ et al (2019) Community richness of amphibian skin bacteria correlates with bioclimate at the global scale. Nat Ecol Evol 33:381–389
Lee HB, Patriarca A, Magan N (2015) Alternaria in food: ecophysiology, mycotoxin production and toxicology. Mycobiology 432:93–106
Lemieux-Labonté V, Simard A, Willis CKR, Lapointe FJ (2017) Enrichment of beneficial bacteria in the skin microbiota of bats persisting with white-nose syndrome. Microbiome 5:115
Lemieux-Labonté V, Tromas N, Shapiro BJ, Lapointe FJ (2016) Environment and host species shape the skin microbiome of captive neotropical bats. PeerJ 4:e2430
Leser TD, Mølbak L (2009) Better living through microbial action: the benefits of the mammalian gastrointestinal microbiota on the host. Environ Microbiol 119:2194–2206
Levy M, Blacher E, Elinav E (2017) Microbiome, metabolites and host immunity. Curr Opin Microbiol 35:8–15
Li A, Li Z, Dai W, Parise KL, Leng H, Jin L, Liu S, Sun K, Hoyt JR, Feng J (2022) Bacterial community dynamics on bats and the implications for pathogen resistance. Environ Microbiol 24:1484–1498
Lorch JM, Palmer JM, Lindner DL et al (2016) First detection of bat white-nose syndrome in western North America. MSphere 14:e00148-e216
Lowrey L, Woodhams DC, Tacchi L, Salinas I (2015) Topographical mapping of the rainbow trout Oncorhynchus mykiss microbiome reveals a diverse bacterial community with antifungal properties in the skin. Appl Environ Microbiol 81:6915–6925
Lutz HL, Jackson EW, Webala PW, Babyesiza WS, KerbisPeterhans JC, Demos TC, Patterson BD, Gilbert JA (2019) Ecology and host identity outweigh evolutionary history in shaping the bat microbiome. Msystems 4:e00511-e519
Moloney RD, Desbonnet L, Clarke G, Dinan TG, Cryan JF (2014) The microbiome: stress, health and disease. Mamm Genome 251:49–74
Marcelino R V, Holmes EC, Sorrell TC (2020) The use of taxon-specific reference databases compromises metagenomic classification. BMC Genomics 21:184
Muth C, Oravecz Z, Gabry J (2018) User-friendly Bayesian regression modeling: a tutorial with Rstanarm and Shinystan. Quant Methods Psychol 14:99–119
Nguyen NH, Song Z, Bates ST et al (2016) FUNGuild: an open annotation tool for parsing fungal community datasets by ecological guild. Fungal Ecol 20:241–248
Nilsson RH, Larsson KH, Taylor AFS et al (2019) The UNITE database for molecular identification of fungi: handling dark taxa and parallel taxonomic classifications. Nucleic Acids Res 47(D1):D259–D264
Njus KA (2014) Molecular techniques for the identification of commensal fungal populations on cave roosting bats (Doctoral dissertation, University of Akron)
Oksanen J, Kindt R, Legendre P, O’Hara B, Stevens MHH, Oksanen MJ, Suggests MASS (2007) The vegan package. Community Ecol Packag 10:631-637 719
Ogórek R et al (2020) A culture-based ID of micromycetes on the wing membranes of greater mouse-eared bats (Myotis Myotis) from the ‘Nietoperek’ site (Poland). Animals 10:1337
Pannkuk EL et al (2013) Sebaceous lipid profiling of bat integumentary tissues: quantitative analysis of free fatty acids, monoacylglycerides, squalene, and sterols. Chem Biodivers 10:2122–2132
Pettit JL, O’Keefe JM (2017) Impacts of white-nose syndrome observed during long-term monitoring of a midwestern bat community. J Fish Wildl Manag 81:69–78
Phillips CD, Hanson J, Wilkinson JE, Koenig L, Rees E, Webala P, Kingston T (2017) Microbiome structural and functional interactions across host dietary niche space. Integr Comp Biol 574:743–755
Pietrangelo L, Bucci A, Maiuro L, Bulgarelli D, Naclerio G (2018) Unraveling the composition of the root-associated bacterial microbiota of Phragmitesaustralis and Typhalatifolia. Front Microbiol 9:1650
R Core Team (2013) “R: A language and environment for statistical computing.” 201
Racey PA (1982) Ecology of bat reproduction p 57–104. In: Kunz TH ed Ecology of bats. Plenum Press New York NY
Reichard JD, Kunz TH (2009) White-nose syndrome inflicts lasting injuries to the wings of little brown myotis Myotislucifugus. Acta Chiropterol 11:457–464
Rideout JR, He Y, Navas-Molina JA et al (2014) Subsampled open-reference clustering creates consistent comprehensive OTU definitions and scales to billions of sequences. PeerJ 2:e545
Rios-Sotelo GR, Northup D, Buecher D, Voyles JL (2018) Skin secretions may provide bats with innate immune defenses against Pseudogymnoascusdestructans. Integr Comp Biol 58:E188
Ross AA, Rodrigues Hoffmann A, Neufeld JD (2019) The skin microbiome of vertebrates. Microbiome 7:1–14
Sisti M, Savini V (2014) Antifungal properties of the human Metschnikowia strain IHEM 25107. Folia Microbiol 593:263–266
Sapkota TB, Jat ML, Aryal JP, Jat RK, Khatri-Chhetri A (2015) Climate change adaptation, greenhouse gas mitigation and economic profitability of conservation agriculture: some examples from cereal systemsof Indo-Gangetic Plains. J Integr Agric 148:1524–1533
Swartz SM, Groves MS, Kim HD, Walsh WR (1996) Mechanical properties of bat wing membrane skin. J Zool 2392:357–378
Thomas T, Moitinho-Silva L, Lurgi M et al (2016) Diversity structure and convergent evolution of the global sponge microbiome. Nat Commun 71:1–12
Thompson L, Sanders J, McDonald D et al (2017) A communal catalogue reveals Earth’s multiscale microbial diversity. Nature 551:457–463
Vanderwolf KJ, Malloch D, McAlpine DF (2015) Fungi associated with over-wintering tricolored bats, Perimyotis subflavus, in a white-nose syndrome region of eastern Canada. J Cave Karst Stud 77:77
Vanderwolf KJ, Campbell LJ, Taylor DR, Goldberg TL, Blehert DS, Lorch JM (2021) Mycobiome traits associated with disease tolerance predict many Western north American bat species will be susceptible to White-nose syndrome. Microbiol Spectr 9:e00254-e321
Vanderwolf KJ et al (2021) Skin fungal assemblages of bats vary based on susceptibility to white-nose syndrome. ISME J 15:909–920
Vanderwolf KJ, McAlpine DF (2021) Hibernacula microclimate and declines in overwintering bats during an outbreak of white-nose syndrome near the northern range limit of infection in North America. Ecol Evol 115:2273–2288
Visagie CM, Neriman YK, Vanderwolf JB, Renaud MW, Sumarah J, Houbraken R, Assebgui K, Seifert D (2020) Penicillium diversity in Canadian bat caves including a new species Pspeluncae. Fungal Systematicsand Evol 5:1
Wang Q, Garrity GM, Tiedje JM, Cole JR (2007) Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl Environ Microbiol 73:5261–5267
Watve MG, Tickoo R, Jog MM, Bhole BD (2001) How many antibiotics are produced by the genus Streptomyces? Arch Microbiol 1765:386–390
Winter AS, Hathaway JJM, Kimble JC, Buecher DC, Valdez EW, Porras-Alfaro A, Young JM, Read KJH, Northup DE (2017) Skin and fur bacterial diversity and community structure on American southwestern bats: effects of habitat geography and bat traits. PeerJ 5:e3944
Woodhams DC, Bletz MC, Becker CG et al (2020) Host-associated microbiomes are predicted by immune system complexity and climate. Genome Biol 21:1–20
Yatsunenko T, Rey FE, Manary MJ et al (2012) Human gut microbiome viewed across age and geography. Nature 4867:222–227
Zepeda Mendoza ML, Xiong Z, Escalera-Zamudio M et al (2018) Hologenomic adaptations underlying the evolution of sanguivory in the common vampire bat. Nat Ecol Evol 24:659–668
Zhang T, Victor TR, Rajkumar SS, Li X, Okoniewski JC, Hicks AC, Davis AD, Broussard K, LaDeau SL, Chaturvedi S, Chaturvedi V (2014) Mycobiome of the bat white nose syndrome affected caves and mines reveals diversity of fungi and local adaptation by the fungal pathogen Pseudogymnoascus (Geomyce s) destructans. PLoS One 9:e108714
Acknowledgements
We are very grateful to Debbie Buecher, Buecher Biological Consulting for her netting and handling of bats in our sites across New Mexico and Arizona under her permits, and to Dr. Ernie Valdez for his assistance in netting and processing bats in the El Malpais Cerro Rendeja II and El Malpais Cerrito Comadre. We are also very grateful to the many field assistants from the various parks and BLM units who assisted with note taking and other tasks, to the Resources staff who worked with us to arrange permits and field support, to Northup Lab personnel who performed DNA extractions, and to MR DNA for sequencing services.
Permits
Permits were approved by the 2014 Arizona and New Mexico Game and Fish Department Scientific Collecting Permit (SP670210, SCI#3423, SCI#3350); National Park Service Scientific Collecting Permit (CAVE-2014-SCI-0012, ELMA-2013-SCI-0005, ELMA-2014-SCI-0001, and PARA-2012-SCI-0003).
Funding
This work was supported by Colorado Plateau Cooperative Ecosystems Studies Unit (CPCESU)—Carlsbad, Caverns National Park (CAVE) Award #P14AC00793, UNM-101; Colorado Plateau Cooperative Ecosystems Studies Unit (CPCESU)—El Malpais National Monument (ELMA) Award #P14AC00588, UNM-99; Colorado Plateau Cooperative Ecosystems Studies Unit (CPCESU)—Grand Canyon Parashant National Monument (PARA) Award #P12AC10812, UNM-80; Fort Stanton Cave Study Project (FSCSP) and Bureau of Land Management Agreement No. 13–0484; T&E, Inc. Award #TE-EAA-01222014; New Mexico Game & Fish Department Share with Wildlife Award #12516000000045; Western National Parks Association. DCW was supported by the National Science Foundation (IOS-1845634).
Author information
Authors and Affiliations
Contributions
ASW and DEN collected and processed all samples in coordination with other local scientists and bat biologists. PJK and DCW performed all sequences/statistical analyses with input from ASW and DEN. All authors contributed to authorship of the manuscript.
Corresponding author
Ethics declarations
Conflict of Interest
The authors declare no competing interests.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Kearns, P.J., Winter, A.S., Woodhams, D.C. et al. The Mycobiome of Bats in the American Southwest Is Structured by Geography, Bat Species, and Behavior. Microb Ecol 86, 1565–1574 (2023). https://doi.org/10.1007/s00248-023-02230-w
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00248-023-02230-w