Abstract
The aim of this study is to investigate the fungal diversity of green turtle nests and to examine phylogenetic relationships among these isolates. During the nesting season, samples of intra-nest sand and failed eggs were collected from 25% of the surviving nests in Sugözü Beaches, which are amongst the most important nesting beaches for endangered green turtles in the Mediterranean. Twenty-three fungi were identified by molecular techniques. Fungal isolates belonged to genera Aspergillus, Emericella, Rhizopus, Actinomucor and Apophysomyces with two undescribed species. Aspergillus variecolor, Aspergillus quadrilinieatus, Aspergillus tubingensis, Rhizopus oryzae, Actinomucor elegans and Apophysomyces variabilis were firstly detected in all sea turtle nests within this study. Our results demonstrate that 36.4% of the nests had fungal contamination. Also hatching success of the nests contaminated by fungi were significantly lower than the uncontaminated nests (P = 0.029). Also, this may represent a threat to marine turtles and a risk for the health of conservation workers. This study is the first molecular phylogenetic study associated with sea turtle nests in the eastern Mediterranean coast and contributes to the wider body of literature on fungal invasion of sea turtle nests with firstly isolated species. These findings are important for improving potential conservation measures for the nest sites.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Alvarez E, Stchigel AM, Cano J, Sutton DA, Fothergill AW, Chander J, Salas V, Rinaldi MG, Guarro J (2010) Molecular phylogenetic diversity of the emerging mucoralean fungus Apophysomyces: proposal of three new species. Rev Iberoam Micol 27:80–89. https://doi.org/10.1016/j.riam.2010.01.006
Al-Zaydani IA, Al-Hakami AM, Joseph MRP, Kassem VM, Almaghrabi MK, Nageeb A, Hamid ME (2015) Aggressive cutaneous zygomycosis caused by Apophysomyces variabilis in an immunocompetent child. Med Mycol Case Rep 10:11–13. https://doi.org/10.1016/j.mmcr.2015.12.003
Barnes GL (1971) Mycoflora of developing peanut pods in Oklahoma. Mycopathologia 45:85–92. https://doi.org/10.1007/BF02059248
Benjamin CR, Hesseltine CW (1957) The genus Actinomucor. Mycologia 49:240–249
Binder U, Maurer E, Lass-Flörl C (2014) Mucormycosis—from the pathogens to the disease. Clin Microbiol Infec 20: 60–66. https://doi.org/10.1111/1469-0691.12566
Canbolat AF, Atatunc K, Candan O, Barcak D (2005) A new green turtle (Chelonia mydas) nesting site in the Mediterranean: Sugözü beaches, Adana (Turkey). II. Mediterranean Conference on Sea Turtles, Antalya, Turkey, p 65
Chou CC, Hwan CH (1994) Effect of ethanol on the hydrolysis of protein and lipid during the ageing of a Chinese fermented soya bean curd-sufu. J Sci Food Agric 66:393–398. https://doi.org/10.1002/jsfa.2740660318
Cruz WP, dela Calvano TP, Griffith ME, White CE, Kim SH, Sutton DA, Thompson EH, Fu J, Wickes BL, Guarro J, Hospenthal DR (2012) Invasive Apophysomyces variabilis infection in a burn patient. J Clin Microbiol 50:2814–2817. https://doi.org/10.1128/JCM.00671-12
Davel G, Featherston P, Fernández A, Abrantes R, Canteros C, Rodero L, Sztern C, Perrotta D (2001) Maxillary sinusitis caused by Actinomucor elegans. J Clin Microbiol 39:740–742. https://doi.org/10.1128/JCM.39.2.740-742.2001
Doygun H, Alphan H (2006) Monitoring urbanization of İskenderun, Turkey, and its negative implications. Environ Monit Assess 114:145–155. https://doi.org/10.1007/s10661-006-2524-0
El-Gindy AA, Saad RR (1990) Fungi of virgin and cultivated soil of Salhiah desert. Egypt Zentralbl Mikrobiol 145:547–551. https://doi.org/10.1016/S0232-4393(11)80061-2
Elshafie A, Al-Bahry SN, AlKindi AY, Ba-Omar T, Mahmoud I (2007) Mycoflora and aflatoxins in soil, eggshells, and failed eggs of Chelonia mydas at Ras Al-Jinz, Oman. Chelonian Conserv Biol 6:267–270. https://doi.org/10.2744/1071-8443(2007)6[267:MAAISE]2.0.CO;2
Farrow WM (1954) Tropical soil fungi. Mycologia 46:632–646
Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791. https://doi.org/10.2307/2408678
Frisvad JC, Larsen TO, Thrane U, Meijer M, Varga J, Samson RA, Nielsen KF (2011) Fumonisin and ochratoxin production in industrial Aspergillus niger strains. PLoS One 6:e23496. https://doi.org/10.1371/journal.pone.0023496
González A, del Palacio A, Cuétara MS, Gómez C, Carabias E, Malo Q (1996) Zygomycosis: review of 16 cases. Enferm Infecc Microbiol Clin 14:233–239
Guarro J, Chander J, Alvarez E, Stchigel AM, Robin K, Dala U, Rani H, Punia RS, Cano JF (2011) Apophysomyces variabilis infections in humans. Emerg Infect Dis 17:134–135. https://doi.org/10.3201/eid1701.101139
Güçlü Ö, Bıyık H, Şahiner A (2010) Mycoflora identified from loggerhead turtle (Caretta caretta) egg shells and nest sand at Fethiye beach. Turkey. Afr J Microbiol Res 4:408–413
Hamann M, Godfrey MH, Seminoff JA et al (2010) Global research priorities for sea turtles: informing management and conservation in the 21st century. Endanger Species Res 11:245–269. https://doi.org/10.3354/esr00279
Ismaiel AA, Papenbrock J (2015) Mycotoxins: producing fungi and mechanisms of phytotoxicity. Agriculture 5:492–537. https://doi.org/10.3390/agriculture5030492
Keene EL (2012) Microorganisms from sand, cloacal fluid, and eggs of Lepidochelys olivacea and standard testing of cloacal fluid antimicrobial properties. Dissertation, Indiana University—Purdue University
Klich MA (2002) Biogeography of Aspergillus species in soil and litter. Mycologia 94:21–27. https://doi.org/10.2307/3761842
Kumar S, Stecher G, Tamura K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33:1870–1874. https://doi.org/10.1093/molbev/msw054
Mahmud A, Lee R, Munfus-McCray D, Kwiatkowski N, Subramanian A, Neofytos D, Carroll K, Zhang SX (2012) Actinomucor elegans as an emerging cause of mucormycosis. J Clin Microbiol 50:1092–1095. https://doi.org/10.1128/JCM.05338-11
Neves MSC, de Melo Moura CC, de Oliveira LG (2015) Mycobiota from the eggs, nests and stillbirths of Eretmochelys imbricata Linneus 1766 (Testudines: Cheloniidae) in Pernambuco State, Brazil. Afr J Microbiol Res 9:1195–1199. https://doi.org/10.5897/AJMR2015.7389
Phillott AD (2004) Penetration of the eggshell and invasion of embryonic tissue by fungi colonising sea turtle eggs. Herpetofauna 34:44–47. https://doi.org/10.1139/z06-125
Phillott AD, Parmenter CJ (2001) The distribution of failed eggs and the appearance of fungi in artificial nests of green (Chelonia mydas) and loggerhead (Caretta caretta) sea turtles. Aust J Zool 49:713–718. https://doi.org/10.1071/ZO00051
Phillott AD, Parmenter CJ (2012) Anti-fungal properties of sea turtle cloacal mucus and egg albumen. Mar Turt Newsl 134:17–21
Phillott AD, Parmenter CJ (2014) Fungal colonization of green sea turtle (Chelonia mydas) nests is unlikely to affect hatchling condition. Herpetol Conserv Biol 9:297–301
Phillott AD, Parmenter CJ, Limpus CJ (2001) Mycoflora identified from failed green (Chelonia mydas) and loggerhead (Caretta caretta) sea turtle eggs at Heron Island, Australia. Chelonian Conserv Biol 4:170–172
Phillott AD, Parmenter CJ, Limpus CJ, Harrower KM (2002) Mycobiota as acute and chronic cloacal contaminants of female sea turtles. Aust J Zool 50:687–695. https://doi.org/10.1071/ZO01057
Phillott AD, Parmenter CJ, Limpus CJ (2004) Occurrence of mycobiota in eastern Australian sea turtle nests. Mem Qld Mus 49:701–703
Prakash H, Ghosh AK, Rudramurthy SM, Paul RA, Gupta S, Negi V, Chakrabarti A (2016) The environmental source of emerging Apophysomyces variabilis infection in India. Med Mycol 54:567–575. https://doi.org/10.1093/mmy/myw014
Rahim S, Dawar S, Tariq M (2010) Mycoflora associated with lentil (Lens culinaris L.) seeds of Pakistan. Pak J Bot 42:4345–4352
Rees AF, Alfaro-Shigueto J, Barata PCR et al (2016) Are we working towards global research priorities for management and conservation of sea turtles? Endanger Species Res 31:337–382. https://doi.org/10.3354/esr00801
Ribes JA, Vanover-Sams CL, Baker DJ (2000) Zygomycetes in human disease. Clin Microbiol Rev 13:236–301. https://doi.org/10.1128/CMR.13.2.236-301.2000
Richardson M (2009) The ecology of the Zygomycetes and its impact on environmental exposure. Clin Microbiol Infect 15:2–9. https://doi.org/10.1111/j.1469-0691.2009.02972.x
Robeck TR, Dalton LM (2002) Saksenaea vasiformis and Apophysomyces elegans zygomycotic infections in bottlenose dolphins (Tursiops truncatus), a killer whale (Orcinus orca), and pacific white-sided dolphins (Lagenorhynchus obliquidens). J Zoo Wildl Med 33:356–366. https://doi.org/10.1638/1042-7260(2002)033[0356:SVAAEZ]2.0
Rosado-Rodríguez G, Maldonado-Ramírez SL (2016) Mycelial fungal diversity associated with the leatherback sea turtle (Dermochelys coriacea) nests from western Puerto Rico. Chelonian Conserv Biol 15:265–272. https://doi.org/10.2744/CCB-1217.1
Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425. https://doi.org/10.1093/oxfordjournals.molbev.a040454
Sarmiento-Ramírez JM, Abella E, Martín MP, Tellería MT, López-Jurado LF, Marco A, Diéguez-Uribeondo J (2010) Fusarium solani is responsible for mass mortalities in nests of loggerhead sea turtle, Caretta caretta, in Boavista, Cape Verde. Fems Microbiol Lett 312:192–200. https://doi.org/10.1111/j.1574-6968.2010.02116.x
Sarmiento-Ramírez JM, Abella-Pérez E, Phillott AD, Sim J, Van West P, Martín MP, Marco A, Diéguez-Uribeondo J (2014) Global distribution of two fungal pathogens threatening endangered sea turtles. PLoS One 9:e85853. https://doi.org/10.1371/journal.pone.0085853
Solomon SE, Baird T (1980) The effect of fungal penetration on the eggshell of the green turtle. In: Seventh European congress on electron microscopy. The Hague, The Netherlands, pp 434–435
Tamura K, Nei M, Kumar S (2004) Prospects for inferring very large phylogenies by using the neighbor-joining method. Proc Natl Acad Sci USA 101:11030–11035. https://doi.org/10.1073/pnas.0404206101
Tedersoo L, Bahram M, Põlme S et al (2014) Global diversity and geography of soil fungi. Science 346:1052–1053. https://doi.org/10.1126/science.1256688
Varga J, Péteri Z, Tábori K, Téren J, Vágvölgyi C (2005) Degradation of ochratoxin A and other mycotoxins by Rhizopus isolates. Int J Food Microbiol 99:321–328. https://doi.org/10.1016/j.ijfoodmicro.2004.10.034
Zhou XX, Zhao DD, Liu JH, Lu F, Ding YT (2014) Physical, chemical and microbiological characteristics of fermented surimi with Actinomucor elegans. LWT Food Sci Technol 59:335–341. https://doi.org/10.1016/j.lwt.2014.05.045
Acknowledgements
I would like to express my very great appreciation to Onur Candan, PhD., for his valuable and constructive suggestions during the planning and development of this research work. I thank Baran Yoğurtçuoğlu, PhD., for comments and suggestions on the earlier version of the manuscript and volunteers of Sea Turtle Conservation Project, which is supported by BIL (BOTAŞ International Limited Co.). And also thanks to Seaturtle.org for Maptool.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Ethical approval
None of the experiments involved sacrificing animals and, therefore, we did not require a specific approval from any institutional animal research ethics committee.
Additional information
Communicated by Erko Stackebrandt.
Rights and permissions
About this article
Cite this article
Candan, E.D. Molecular identification of fungal isolates and hatching success of green turtle (Chelonia mydas) nests. Arch Microbiol 200, 911–919 (2018). https://doi.org/10.1007/s00203-018-1496-0
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00203-018-1496-0