Isolation and identification of fungi, collected from Proteus anguinus and its immediate aquatic environment

Dvoživke, vključno s človeško ribico, spadajo med ogrožene skupine živali v svetovnem merilu. Med glavne grožnje spadajo uničevanje in onesnaževanje njihovega naravnega habitata ter podnebne spremembe. Glede na skromne podatke o patogenih človeške ribice v ujetništvu in prosto živečih populacijah, smo v diplomski nalogi želeli ugotovili, če dodatno grožnjo predstavljajo tudi glive. Zanimala nas je pojavnost oportuno patogenih gliv, ki so bile nedavno odkrite kot povzročiteljice hitridiomikoz, mukormikoz, kromomikoz in saproleginoz. Poleg tega pa smo želeli ugotoviti prisotnost gliv v kožnem mikrobiomu človeške ribice, saj o tem ni podatkov. Tekom diplomske naloge smo izolirali glive iz neposrednega vodnega okolja človeške ribice v naravi kot tudi v ujetništvu. Na lokacijah Jelševnik, Krška in Planinska jama smo vzorčili vodo, ter v njej nastavili vabe. Iz istih lokacij in dodatno iz Postojnske jame smo jemali brise površine živali, tako zdravih kot bolnih, in jih gojili na različnih gojiščih pri 15 °C in 20 °C. Iz vseh vzorcev smo osamili 113 glivnih sevov, ki smo jih na podlagi nukleotidnih zaporedij molekularnih označevalcev, predvsem ITS rDNA, uvrstili v 43 rodov. V umetnem in naravnem okolju človeške ribice se nahajajo potencialno nevarne glive rodov: Aspergillus, Candida, Cladosporium, Exophiala, Mucor in Saprolegnia, ki so znani povzročitelji bolezni dvoživk. V vzorcih vode, kot tudi na mikrobioloških vabah smo opazili pojavnost istih rodov gliv, prevladovala sta rodova Mucor in Trichoderma. Nabor gliv izoliranih s površine proteusov se je delno prekrival s tistimi iz okolja, vendar pa je bila ponovljivost izolacije istih rodov pri različnih osebkih zelo majhna, prav tako so se kolonije gliv pojavljale sporadično. Opazili smo razlike v pojavnosti rodov gliv izoliranih z zdravih in bolnih osebkov. Pri zdravih osebkih so se na koži največkrat pojavljale glive rodov Exophiala in Cladosporium (tri živali), pri bolnih pa Saprolegnia (2 živali). Vrste rodu Candida so se pojavljale pri dveh zdravih in eni bolnih živali izključno v ujetništvu.Amphibians, including human fish, belong on a global scale to endangered groups of animals. The main threats include the destruction and pollution of their natural habitats and climate change. Considering the modest data of human fish pathogens in captivity and wild populations, we wanted to find out whether the fungi are also an additional threat. We were interested in the incidence of opportunistic pathogens that were recently detected as the causative agents of hitridiomycosis, mucormycosis, chromomycosis and saproleginosis. In addition, since there is no data available, we wanted to determine the presence of fungi in human fish microbial skin. During the diploma we isolated fungi from the immediate water environment of the human fish in nature and in captivity. At the locations such as Jelševnik, Krška and Planinska jama we sampled the water and we set the microbiological baits there. From the same locations, and additionally from Postojna Cave, we were taking samples from healthy and sick animals’ surfaces, and they were grown on different media at 15 ° C and 20 ° C. All samples were isolated from 113 fungal strains, which were classified into 43 genera on the basis of nucleotide sequences of molecular markers, especially ITS rDNA. In the artificial and natural environment of the human fish there are potentially dangerous fungi of the genera: Aspergillus, Candida, Cladosporium, Exophiala, Mucor and Saprolegnia, known as causative agents of diseases of amphibians. In the water samples, as well as on microbiological baits, the appearance of the same fungus species was observed, with the genus Mucor and Trichoderma predominant. The set of samples isolated from the surface of proteus were partially similar to those taken from the environment. However, the reproducibility of isolation of the same genera in the various animals was very small, and the fungal colonies appeared sporadically. We observed differences in the incidence of fungal species isolated from healthy and sick animals. With healthy animals, the most commonly occurring genera were Exophila and Cladosporium (three animals), and with sick animals Saprolegnia (2 animals). Candida species appeared with two healthy and one sick animal exclusively in captivity