Environmental factors and host sex influence the skin microbiota structure of Hong Kong newt (Paramesotriton hongkongensis) in a coldspot of chytridiomycosis in subtropical East Asia

Bowen WAN , Guoling CHEN , Emily Shui Kei POON , Hon Shing FUNG , Anthony LAU , Simon Yung Wa SIN

Integrative Zoology ›› 2025, Vol. 20 ›› Issue (2) : 236 -255.

PDF
Integrative Zoology ›› 2025, Vol. 20 ›› Issue (2) : 236 -255. DOI: 10.1111/1749-4877.12855
ORIGINAL ARTICLE

Environmental factors and host sex influence the skin microbiota structure of Hong Kong newt (Paramesotriton hongkongensis) in a coldspot of chytridiomycosis in subtropical East Asia

Author information +
History +
PDF

Abstract

Chytridiomycosis, an infectious skin disease caused by the chytrid fungi, Batrachochytrium dendrobatidis and B. salamandrivorans, poses a significant threat to amphibian biodiversity worldwide. Antifungal bacteria found on the skin of chytrid-resistant amphibians could potentially provide defense against chytridiomycosis and lower mortality rates among resistant individuals. The Hong Kong newt (Paramesotriton hongkongensis) is native to East Asia, a region suspected to be the origin of chytrids, and has exhibited asymptomatic infection, suggesting a long-term coexistence with the chytrids. Therefore, the skin microbiota of this resistant species warrant investigation, along with other factors that can affect the microbiota. Among the 149 newts sampled in their natural habitats in Hong Kong, China, putative antifungal bacteria were found in all individuals. There were 314 amplicon sequence variants distributed over 25 genera of putative antifungal bacteria; abundant ones included Acinetobacter, Flavobacterium, and Novosphingobium spp. The skin microbiota compositions were strongly influenced by the inter-site geographical distances. Despite inter-site differences, we identified some core skin microbes across sites that could be vital to P. hongkongensis. The dominant cores included the family Comamonadaceae, family Chitinophagaceae, and class Betaproteobacteria. Moreover, habitat elevation and host sex also exhibited significant effects on skin microbiota compositions. The antifungal bacteria found on these newts offer an important resource for conservation against chytridiomycosis, such as developing probiotic treatments for susceptible species.

Keywords

16S amplicon sequencing / antifungal bacteria / Batrachochytrium dendrobatidis / Batrachochytrium salamandrivorans / host–pathogen–microbiota coevolution

Cite this article

Download citation ▾
Bowen WAN, Guoling CHEN, Emily Shui Kei POON, Hon Shing FUNG, Anthony LAU, Simon Yung Wa SIN. Environmental factors and host sex influence the skin microbiota structure of Hong Kong newt (Paramesotriton hongkongensis) in a coldspot of chytridiomycosis in subtropical East Asia. Integrative Zoology, 2025, 20(2): 236-255 DOI:10.1111/1749-4877.12855

登录浏览全文

4963

注册一个新账户 忘记密码

References

[1]

Abarca JG, Zuniga I, Ortiz-Morales G et al. (2018). Characterization of the skin microbiota of the cane toad Rhinella cf. marina in Puerto Rico and Costa Rica. Frontiers in Microbiology 8, 2624.

[2]

Alberdi A, Gilbert MTP (2019). hilldiv: An R package for the integral analysis of diversity based on Hill numbers. bioRxiv545665. https://doi.org/10.1101/545665

[3]

Andrews S (2010). FastQC: A quality control tool for high throughput sequence data. Available from URL: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/

[4]

Antwis RE, Haworth RL, Engelmoer DJ, Ogilvy V, Fidgett AL, Preziosi RF (2014). Ex situ diet influences the bacterial community associated with the skin of red-eyed tree frogs (Agalychnis callidryas). PLoS ONE 9, e85563.

[5]

Apprill A, Mcnally S, Parsons R, Weber L (2015). Minor revision to V4 region SSU rRNA 806R gene primer greatly increases detection of SAR11 bacterioplankton. Aquatic Microbial Ecology 75, 129-137.

[6]

Bataille A, Fong JJ, Cha M et al. (2013). Genetic evidence for a high diversity and wide distribution of endemic strains of the pathogenic chytrid fungus Batrachochytrium dendrobatidis in wild A sian amphibians. Molecular Ecology 22, 4196-4209.

[7]

Bataille A, Lee-Cruz L, Tripathi B, Kim H, Waldman B (2016). Microbiome variation across amphibian skin regions: Implications for chytridiomycosis mitigation efforts. Microbial Ecology 71, 221-232.

[8]

Bataille A, Lee-Cruz L, Tripathi B, Waldman B (2018). Skin bacterial community reorganization following metamorphosis of the fire-bellied toad (Bombina orientalis). Microbial Ecology 75, 505-514.

[9]

Bates KA, Clare FC, O'hanlon S et al. (2018). Amphibian chytridiomycosis outbreak dynamics are linked with host skin bacterial community structure. Nature Communications 9, 693.

[10]

Bates KA, Shelton JM, Mercier VL et al. (2019). Captivity and infection by the fungal pathogen Batrachochytrium salamandrivorans perturb the amphibian skin microbiome. Frontiers in Microbiology 10, 1834.

[11]

Becker MH, Brucker RM, Schwantes CR, Harris RN, Minbiole KP (2009). The bacterially produced metabolite violacein is associated with survival of amphibians infected with a lethal fungus. Applied and Environmental Microbiology 75, 6635-6638.

[12]

Belden LK, Hughey MC, Rebollar EA et al. (2015). Panamanian frog species host unique skin bacterial communities. Frontiers in Microbiology 6, 1171.

[13]

Bird AK, Prado-Irwin SR, Vredenburg VT, Zink AG (2018). Skin microbiomes of California terrestrial salamanders are influenced by habitat more than host phylogeny. Frontiers in Microbiology 9, 442.

[14]

Bletz MC, Archer H, Harris RN et al. (2017a). Host ecology rather than host phylogeny drives amphibian skin microbial community structure in the biodiversity hotspot of Madagascar. Frontiers in Microbiology 8, 1530.

[15]

Bletz MC, Kelly M, Sabino-Pinto J et al. (2018). Disruption of skin microbiota contributes to salamander disease. Proceedings of the Royal Society B 285, 20180758.

[16]

Bletz MC, Perl RB, Bobowsk BT et al. (2017c). Amphibian skin microbiota exhibits temporal variation in community structure but stability of predicted Bd-inhibitory function. The ISME Journal 11, 1521-1534.

[17]

Bletz MC, Vences M, Sabino-Pinto J et al. (2017b). Cutaneous microbiota of the Japanese giant salamander (Andrias japonicus), a representative of an ancient amphibian clade. Hydrobiologia 795, 153-167.

[18]

Bresciano J, Salvador C, Paz-Y-Mino C et al. (2015). Variation in the presence of anti-Batrachochytrium dendrobatidis bacteria of amphibians across life stages and elevations in Ecuador. Ecohealth 12, 310-319.

[19]

Bright M, Bulgheresi S (2010). A complex journey: Transmission of microbial symbionts. Nature Reviews Microbiology 8, 218-230.

[20]

Brooks AW, Kohl KD, Brucker RM, Van Opstal EJ, Bordenstein SR (2016). Phylosymbiosis: Relationships and functional effects of microbial communities across host evolutionary history. PLoS Biology 14, e2000225.

[21]

Brucker RM, Harris RN, Schwantes CR et al. (2008). Amphibian chemical defense: Antifungal metabolites of the microsymbiont Janthinobacterium lividum on the salamander Plethodon cinereus. Journal of Chemical Ecology 34, 1422-1429.

[22]

Brunetti AE, Lyra ML, Bauermeister A et al. (2023). Host macrocyclic acylcarnitines mediate symbiotic interactions between frogs and their skin microbiome. Iscience 26, 108109.

[23]

Cammarota G, Ianiro G, Gasbarrini A (2014). Fecal microbiota transplantation for the treatment of Clostridium difficile infection: a systematic review. Journal of Clinical Gastroenterology 48, 693-702.

[24]

Chen G, Lau A, Wan B et al. (2023). Occurrence of pathogenic chytrid fungi Batrachochytrium salamandrivorans and Batrachochytrium dendrobatidis in the Hong Kong newt (Paramesotriton hongkongensis) and other wild and imported amphibians in a subtropical Asian region. The Journal of Wildlife Diseases 59, 709-721.

[25]

Chen YE, Tsao H (2013). The skin microbiome: Current perspectives and future challenges. Journal of the American Academy of Dermatology 69, 143-155. e3.

[26]

Colston TJ, Jackson CR (2016). Microbiome evolution along divergent branches of the vertebrate tree of life: What is known and unknown. Molecular Ecology 25, 3776-3800.

[27]

Crawford AJ, Lips KR, Bermingham E (2010). Epidemic disease decimates amphibian abundance, species diversity, and evolutionary history in the highlands of central Panama. PNAS 107, 13777-13782.

[28]

Cruaud P, Rasplus J-Y, Rodriguez LJ, Cruaud A (2017). High-throughput sequencing of multiple amplicons for barcoding and integrative taxonomy. Scientific Reports 7, 41948.

[29]

Douglas AJ, Hug LA, Katzenback BA (2021). Composition of the North American wood frog (Rana sylvatica) bacterial skin microbiome and seasonal variation in community structure. Microbial Ecology 81, 78-92.

[30]

Edgar RC (2010). Search and clustering orders of magnitude faster than BLAST. Bioinformatics 26, 2460-2461.

[31]

Edgar RC (2016a). SINTAX: A simple non-Bayesian taxonomy classifier for 16S and ITS sequences. bioRxiv074161.

[32]

Edgar RC (2016b). UNOISE2: Improved error-correction for Illumina 16S and ITS amplicon sequencing. bioRxiv081257.

[33]

Edgar RC, Flyvbjerg H (2015). Error filtering, pair assembly and error correction for next-generation sequencing reads. Bioinformatics 31, 3476-3482.

[34]

Escallon Herkrath C (2015). Reproductive physiology, avian malaria, and the cloacal microbiome in tropical Rufous-collared Sparrows (Zonotrichia capensis) (Dissertation). Virginia Tech, Blacksburg, USA.

[35]

Fierer N, Jackson RB (2006). The diversity and biogeography of soil bacterial communities. PNAS 103, 626-631.

[36]

Fisher MC, Garner TW (2020). Chytrid fungi and global amphibian declines. Nature Reviews Microbiology 18, 332-343.

[37]

Foster KR, Schluter J, Coyte KZ, Rakoff-Nahoum S (2017). The evolution of the host microbiome as an ecosystem on a leash. Nature 548, 43-51.

[38]

Fredricks DN (2001). Microbial ecology of human skin in health and disease. Journal of Investigative Dermatology Symposium Proceedings 6, 167-169.

[39]

Fu VWK, Karraker NE, Dudgeon D (2013). Breeding dynamics, diet, and body condition of the Hong Kong newt (Paramesotriton hongkongensis). Herpetological Monographs 27, 1-22.

[40]

Galili T (2015). dendextend: An R package for visualizing, adjusting and comparing trees of hierarchical clustering. Bioinformatics 31, 3718-3720.

[41]

Grice EA, Segre JA (2011). The skin microbiome. Nature Reviews Microbiology 9, 244-253.

[42]

Harris RN, Brucker RM, Walke JB et al. (2009a). Skin microbes on frogs prevent morbidity and mortality caused by a lethal skin fungus. The ISME Journal 3, 818-824.

[43]

Harris RN, James TY, Lauer A, Simon MA, Patel A (2006). Amphibian pathogen Batrachochytrium dendrobatidis is inhibited by the cutaneous bacteria of amphibian species. EcoHealth 3, 53-56.

[44]

Harris RN, Lauer A, Simon MA, Banning JL, Alford RA (2009b). Addition of antifungal skin bacteria to salamanders ameliorates the effects of chytridiomycosis. Diseases of Aquatic Organisms 83, 11-16.

[45]

Harrison XA, Price SJ, Hopkins K, Leung WT, Sergeant C, Garner TW (2019). Diversity-stability dynamics of the amphibian skin microbiome and susceptibility to a lethal viral pathogen. Frontiers in Microbiology 10, 2883.

[46]

Hill MO (1973). Diversity and evenness: A unifying notation and its consequences. Ecology 54, 427-432.

[47]

Hoffmann AR (2017). The cutaneous ecosystem: The roles of the skin microbiome in health and its association with inflammatory skin conditions in humans and animals. Advances in Veterinary Dermatology 8, 71-83.

[48]

Huang PY, Poon ESK, Chan LY et al. (2022). Dietary diversity of multiple shorebird species in an Asian subtropical wetland unveiled by DNA metabarcoding. Environmental DNA 4, 1381-1396.

[49]

Huang P-Y, Poon ESK, Wong ATC, So IWY, Sung Y-H, Sin SYW (2021). DNA metabarcoding reveals the dietary composition in the endangered black-faced spoonbill. Scientific Reports 11, 18773.

[50]

Hughey MC, Pena JA, Reyes R, Medina D, Belden LK, Burrowes PA (2017). Skin bacterial microbiome of a generalist Puerto Rican frog varies along elevation and land use gradients. PeerJ 5, e3688.

[51]

IUCN (2021). The IUCN Red List of Threatened Species. Version 2021-3. Available online atfrom URL: https://www.iucnredlist.org

[52]

Jani AJ, Briggs CJ (2014). The pathogen Batrachochytrium dendrobatidis disturbs the frog skin microbiome during a natural epidemic and experimental infection. PNAS 111, E5049-E5058.

[53]

Jiménez RR, Alvarado G, Estrella J, Sommer S (2019). Moving beyond the host: Unraveling the skin microbiome of endangered Costa Rican amphibians. Frontiers in Microbiology 10, 2060.

[54]

Krynak KL, Burke DJ, Benard MF (2016). Landscape and water characteristics correlate with immune defense traits across Blanchard's cricket frog (Acris blanchardi) populations. Biological Conservation 193, 153-167.

[55]

Kueneman JG, Parfrey LW, Woodhams DC, Archer HM, Knight R, Mckenzie VJ (2014). The amphibian skin-associated microbiome across species, space and life history stages. Molecular Ecology 23, 1238-1250.

[56]

Kueneman JG, Weiss S, Mckenzie VJ (2017). Composition of micro-eukaryotes on the skin of the cascades frog (Rana cascadae) and patterns of correlation between skin microbes and Batrachochytrium dendrobatidis. Frontiers in Microbiology 8, 2350.

[57]

Kueneman JG, Woodhams DC, Harris R, Archer HM, Knight R, Mckenzie VJ (2016). Probiotic treatment restores protection against lethal fungal infection lost during amphibian captivity. Proceedings of the Royal Society B: Biological Sciences 283, 20161553.

[58]

Küng D, Bigler L, Davis LR, Gratwicke B, Griffith E, Woodhams DC (2014). Stability of microbiota facilitated by host immune regulation: informing probiotic strategies to manage amphibian disease. PLoS ONE 9, e87101.

[59]

Lai Y, Cogen AL, Radek KA et al. (2010). Activation of TLR2 by a small molecule produced by Staphylococcus epidermidis increases antimicrobial defense against bacterial skin infections. Journal of Investigative Dermatology 130, 2211-2221.

[60]

Laking AE, Ngo HN, Pasmans F, Martel A, Nguyen TT (2017). Batrachochytrium salamandrivorans is the predominant chytrid fungus in Vietnamese salamanders. Scientific Reports 7, 44443.

[61]

Lau A (2017). Conservation and population biology of the Hong Kong newt (Paramesotriton hongkongensis) (Thesis). University of Hong Kong, Hong Kong, China.

[62]

Lau A, Karraker NE, Martelli P, Dudgeon D (2017). Delineation of core terrestrial habitat for conservation of a tropical salamander: The Hong Kong newt (Paramesotriton hongkongensis). Biological Conservation 209, 76-82.

[63]

Lauer A, Simon MA, Banning JL, Lam BA, Harris RN (2008). Diversity of cutaneous bacteria with antifungal activity isolated from female four-toed salamanders. The ISME Journal 2, 145-157.

[64]

Loudon AH, Woodhams DC, Parfrey LW et al. (2014). Microbial community dynamics and effect of environmental microbial reservoirs on red-backed salamanders (Plethodon cinereus). The ISME Journal 8, 830-840.

[65]

Mallott EK, Borries C, Koenig A, Amato KR, Lu A (2020). Reproductive hormones mediate changes in the gut microbiome during pregnancy and lactation in Phayre's leaf monkeys. Scientific Reports 10, 9961.

[66]

Martel A, Blooi M, Adriaensen C et al. (2014). Recent introduction of a chytrid fungus endangers Western Palearctic salamanders. Science 346, 630-631.

[67]

Martel A, Spitzen-van der Sluijs A, Blooi M et al. (2013). Batrachochytrium salamandrivorans sp. nov. causes lethal chytridiomycosis in amphibians. PNAS 110, 15325-15329.

[68]

Martin M (2011). Cutadapt removes adapter sequences from high-throughput sequencing reads. EMBnet. Journal 17, 10-12.

[69]

Martinez Arbizu P (2020). pairwiseAdonis: pairwise multilevel comparison using adonis. R package version 0.4. Available from URL: https://github.com/pmartinezarbizu/pairwiseAdonis

[70]

Martín-Platero AM, Valdivia E, Ruíz-Rodríguez M et al. (2006). Characterization of antimicrobial substances produced by Enterococcus faecalis MRR 10-3, isolated from the uropygial gland of the hoopoe (Upupa epops). Applied and Environmental Microbiology 72, 4245-4249.

[71]

Martín-Vivaldi M, Pena A, Peralta-Sánchez JM et al. (2010). Antimicrobial chemicals in hoopoe preen secretions are produced by symbiotic bacteria. Proceedings of the Royal Society B: Biological Sciences 277, 123-130.

[72]

Mckenzie VJ, Bowers RM, Fierer N, Knight R, Lauber CL (2012). Co-habiting amphibian species harbor unique skin bacterial communities in wild populations. The ISME Journal 6, 588-596.

[73]

Mcmurdie PJ, Holmes S (2013). phyloseq: An R package for reproducible interactive analysis and graphics of microbiome census data. PLoS ONE 8, e61217.

[74]

Muletz-Wolz CR, Direnzo GV, Yarwood SA, Campbell Grant EH, Fleischer RC, Lips KR (2017). Antifungal bacteria on woodland salamander skin exhibit high taxonomic diversity and geographic variability. Applied and Environmental Microbiology 83, e00186-17.

[75]

Muletz-Wolz CR, Fleischer RC, Lips KR (2019). Fungal disease and temperature alter skin microbiome structure in an experimental salamander system. Molecular Ecology 28, 2917-2931.

[76]

Muletz Wolz CR, Yarwood SA, Campbell Grant EH, Fleischer RC, Lips KR (2018). Effects of host species and environment on the skin microbiome of Plethodontid salamanders. Journal of Animal Ecology 87, 341-353.

[77]

Mutnale MC, Reddy GS, Vasudevan K (2021). Bacterial community in the skin microbiome of frogs in a coldspot of chytridiomycosis infection. Microbial Ecology 82, 554-558.

[78]

Naik S, Bouladoux N, Linehan JL et al. (2015). Commensal-dendritic-cell interaction specifies a unique protective skin immune signature. Nature 520, 104-108.

[79]

O'hanlon SJ, Rieux A, Farrer RA et al. (2018). Recent Asian origin of chytrid fungi causing global amphibian declines. Science 360, 621-627.

[80]

Oksanen J, Simpson G, Blanchet FG et al. (2022). Vegan: community ecology package, Version 2.6-2. Vienna (Austria): R Foundation for Statistical Computing. Available from URL: https://github.com/vegandevs/vegan

[81]

Parada AE, Needham DM, Fuhrman JA (2016). Every base matters: Assessing small subunit rRNA primers for marine microbiomes with mock communities, time series and global field samples. Environmental Microbiology 18, 1403-1414.

[82]

Pérez-Sánchez T, Balcázar J, García Y et al. (2011). Identification and characterization of lactic acid bacteria isolated from rainbow trout, Oncorhynchus mykiss (Walbaum), with inhibitory activity against Lactococcus garvieae. Journal of Fish Diseases 34, 499-507.

[83]

Prado-Irwin SR, Bird AK, Zink AG, Vredenburg VT (2017). Intraspecific variation in the skin-associated microbiome of a terrestrial salamander. Microbial Ecology 74, 745-756.

[84]

Prest TL, Kimball AK, Kueneman JG, Mckenzie VJ (2018). Host-associated bacterial community succession during amphibian development. Molecular Ecology 27, 1992-2006.

[85]

R Development Core Team (2021). R: A Language and Environment for Statistical Computing. R Foundation for Statistical Computing, Vienna, Austria.

[86]

Rebolla EA, Martinez-Ugalde E, Orta AH (2020). The amphibian skin microbiome and its protective role against chytridiomycosis. Herpetologica 76, 167-177.

[87]

Rognes T, Flouri T, Nichols B, Quince C, Mahé F (2016). VSEARCH: A versatile open source tool for metagenomics. PeerJ 4, e2584.

[88]

Ross AA, Rodrigues HA, Neufeld JD (2019). The skin microbiome of vertebrates. Microbiome 7, 79.

[89]

Roth T, Foley J, Worth J, Piovia-Scott J, Pope K, Lawler S (2013). Bacterial flora on Cascades frogs in the Klamath mountains of California. Comparative Immunology, Microbiology and Infectious Diseases 36, 591-598.

[90]

Sabino-Pinto J, Bletz MC, Islam MM et al. (2016). Composition of the cutaneous bacterial community in Japanese amphibians: Effects of captivity, host species, and body region. Microbial Ecology 72, 460-469.

[91]

Scheele BC, Pasmans F, Skerratt LF et al. (2019). Amphibian fungal panzootic causes catastrophic and ongoing loss of biodiversity. Science 363, 1459-1463.

[92]

Spitzen-Van Der Sluijs A, Spikmans F, Bosman W et al. (2013). Rapid enigmatic decline drives the fire salamander (Salamandra salamandra) to the edge of extinction in the Netherlands. Amphibia-Reptilia 34, 233-239.

[93]

Swei A, Rowley JJ, Rödder D et al. (2011). Is chytridiomycosis an emerging infectious disease in Asia? PLoS ONE 6, e23179.

[94]

Vaelli PM, Theis KR, Williams JE, O'connell LA, Foster JA, Eisthen HL (2020). The skin microbiome facilitates adaptive tetrodotoxin production in poisonous newts. eLife 9, e53898.

[95]

Varga JF, Bui-Marinos MP, Katzenback BA (2019). Frog skin innate immune defences: Sensing and surviving pathogens. Frontiers in Immunology 9, 3128.

[96]

Walke JB, Becker MH, Krinos A et al. (2021). Seasonal changes and the unexpected impact of environmental disturbance on skin bacteria of individual amphibians in a natural habitat. FEMS Microbiology Ecology 97, fiaa248.

[97]

Walker DM, Hill AJ, Albecker MA et al. (2020). Variation in the slimy salamander (Plethodon spp.) skin and gut-microbial assemblages is explained by geographic distance and host affinity. Microbial Ecology 79, 985-997.

[98]

Woodhams DC, Alford RA, Antwis RE et al. (2015). Antifungal isolates database of amphibian skin-associated bacteria and function against emerging fungal pathogens. Ecology 96, 595-595.

[99]

Woodhams DC, Vredenburg VT, Simon M-A et al. (2007). Symbiotic bacteria contribute to innate immune defenses of the threatened mountain yellow-legged frog, Rana muscosa. Biological Conservation 138, 390-398.

[100]

Yang X, Hou X, Wei L et al. (2020). Characterization of skin symbiotic bacteria of sympatric amphibians in Southeastern China. Asian Herpetological Research 11, 381-393B.

[101]

Yoon S-H, Ha S-M, Kwon S et al. (2017). Introducing EzBioCloud: A taxonomically united database of 16S rRNA gene sequences and whole-genome assemblies. International Journal of Systematic and Evolutionary Microbiology 67, 1613.

[102]

Yotsu M, Iorizzi M, Yasumoto T (1990). Distribution of tetrodotoxin, 6-epitetrodotoxin, and 11-deoxytetrodotoxin in newts. Toxicon 28, 238-241.

[103]

Yuan Z, Martel A, Wu J, Van Praet S, Canessa S, Pasmans F (2018). Widespread occurrence of an emerging fungal pathogen in heavily traded Chinese urodelan species. Conservation Letters 11, e12436.

RIGHTS & PERMISSIONS

2024 The Author(s). Integrative Zoology published by International Society of Zoological Sciences, Institute of Zoology/Chinese Academy of Sciences and John Wiley & Sons Australia, Ltd.

AI Summary AI Mindmap
PDF

5

Accesses

0

Citation

Detail

Sections
Recommended

AI思维导图

/