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Izolacija aktinomicet iz slovenskih kraških jam in vrednotenje njihove protimikrobne aktivnosti
ID Vlaj, Doroteja (Author), ID Klančnik, Anja (Mentor) More about this mentor... This link opens in a new window, ID Paš, Maja (Comentor)

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Abstract
Problem zdravljenja nalezljivih bolezni se zaradi pojava odpornosti bakterij povečuje. Razvoj novih protimikrobnih učinkovin se je usmeril v spremembe struktur že obstoječih antibiotikov in v iskanje novih učinkovin ter novih načinov delovanja antibiotikov v celici. Nove učinkovine so lahko prisotne tudi v ekstremnih in premalo raziskanih okoljih, kot so jamski ekosistemi z genetsko in metabolno raznolikimi mikroorganizmi. Zato smo se v magistrskem delu usmerili na jamsko okolje, iz katerega smo v sodelovanju pridobili jamske vzorce sedimentov, prsti, jamskega mleka ter glinene zapolnitve iz vrtin. Nato smo s pomočjo različnih selektivnih gojišč iz jamskih vzorcev pridobili 214 izolatov aktinomicet. Vsak izolat smo gojili v produkcijskem gojišču, iz katerega smo nato pripravili izvleček sekundarnih metabolitov. Pripravljenim izvlečkom sekundarnih metabolitov vsakega izolata aktinomicet smo ovrednotili protimikrobno aktivnost proti bakterijam Micrococcus luteus, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, ter kvasovkam Saccharomyces cerevisiae in Candida albicans. Uporabili smo metodo difuzije v trdnem gojišču z merjenjem con inhibicije. Rezultati so potrdili protimikrobno aktivnost pri 107-ih izvlečkih sekundarnih metabolitov izolatov aktinomicet iz jamskega okolja. Največ izvlečkov (48 %) je imelo protibakterijsko aktivnost proti bakterijami M. luteus. Izpostavimo lahko še protibakterijsko aktivnost proti bakterijam S. aureus (14 % izvlečkov), proti E. coli (7 % izvlečkov), ter proti P. aeruginosa (2 % izvlečkov). Največ izvlečkov (11 %) je imelo protiglivno aktivnost proti kvasovkam S. cerevisiae, izpostavimo lahko še aktivnost proti C. albicans (5 % izvlečkov). Aktivne sekundarne učinkovine izolatov z dokazano protimikrobno aktivnostjo bodo v nadaljevanju raziskav okarakterizirane.

Language:Slovenian
Keywords:aktinomicete, slovenske kraške jame, izolacija, gojenje, ekstrakcija, tankoplastna kromatografija, sekundarni metaboliti, protimikrobna aktivnost
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Place of publishing:Ljubljana
Publisher:[D. Vlaj]
Year:2022
PID:20.500.12556/RUL-134790 This link opens in a new window
UDC:579.87:551.435.84:615.332
COBISS.SI-ID:97993731 This link opens in a new window
Publication date in RUL:02.02.2022
Views:1146
Downloads:148
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Secondary language

Language:English
Title:Isolation of actinomycetes from Slovenian Karst caves and evaluation of their antimicrobial activity
Abstract:
Infectious diseases have recently become a serious threat to human health, due to the emergence of multiple drug resistance. The development of new antimicrobial agents has focused on modifying the structures of existing antibiotics and finding new mechanisms of action, as well as finding new antimicrobial agents in extreme and under-researched environments, such as cave ecosystems that offer unexplored genetic and metabolic diversity. Therefore, in the master's thesis we focused on the cave environment, from which we obtained cave samples of sediments, soil, cave milk and clay fillings from wells. We succeeded in isolating cave actinomycetes using selective media and obtaining extracts of secondary metabolites by culturing strains in a production medium. The extracts were used to test antimicrobial activity against selected bacteria Micrococcus luteus, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and yeast Saccharomyces cerevisiae and Candida albicans. We found that 107 strains of isolated actinomycetes produced bioactive substances that inhibited the growth of test microorganisms. Activity against M. luteus was shown by 48% of the extracts, activity against S. aureus by 14% of the extracts, against E. coli by 7% of the extracts, and against P. aeruginosa by 2% of the extracts. Activity against S. cerevisiae showed 11% of the extracts and against C. albicans 5% of the extracts. Isolated strains of actinomycetes that have shown strong antimicrobial activity against Gram-positive and negative bacteria will be involved in further research with the aim of characterizing pure active substance.

Keywords:actinomycetes, Slovenian Karst caves, isolation, cultivation, extraction, thin layer chromatography, secondary metabolites, antimicrobial activity

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