Razvoj krioprezervacije vodene plijesni Saprolegnia parasitica primjenom eutektičkih otapala

undergraduate thesis

• Title (english): Development of cryopreservation of water mold Saprolegnia parasitica using eutectic solvents
• Author: Sara Delimar
• Mentor: Ana Bielen (mentor)
• Mentor: Marina Cvjetko Bubalo (pomoć pri izradi, neposredni voditelj)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Biology and Microbial Genetics); Zagreb
• Defense date and country: 2024-07-10, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Postojeći protokoli za krioprezervaciju vodene plijesni Saprolegnia parasitica koriste tradicionalne krioprotektante koji mogu biti toksični, a preživljenje nakon odmrzavanja je često suboptimalno. Zbog toga je istražena mogućnost korištenja niskotemperaturnih eutektičkih otapala (engl. Deep Eutectic Solvents, DES) kao alternativnih otapala za krioprezervaciju ovog mikroorganizma. Analizirano je preživljenje i brzina rasta micelija S. parasitica nakon sedmodnevnog zamrzavanja pri -80 °C uz korištenje glicerola kao standarda i 12 DES-ova, od kojih je najbolje rezultate dao glicerol:trehaloza. Predinkubacija u trajanju od 30 minuta uzrokovala je bolji rast nakon odmrzavanja nego duža predinkubacija (1 sat, 3 sata). Postepeno zamrzavanje, nakon predinkubacije od 1 sat, nije imalo značajan učinak na uspješnost krioprezervacije. Iako je u svim pokusima uz korištenje glicerola i DES-a glicerol:trehaloza kao krioprotektanata preživljenje vodene plijesni S. parasitica nakon odmrzavanja bilo 100%, duže vrijeme zamrzavanja (32 dana naspram 7 dana) uzrokovalo je sporiju regeneraciju micelija pa je potrebno dodatno optimirani protokol krioprezervacije vodene plijesni S. parasitica.

Abstract (english)

Existing protocols for cryopreservation of the water mould Saprolegnia parasitica use traditional cryoprotectants that can be toxic and survival after thawing is often suboptimal. Thus, Deep Eutectic Solvents (DES) were investigated as alternative solvents for cryopreservation of this microorganism. The survival and growth rate of S. parasitica mycelium after seven days of freezing at -80 °C were analysed using glycerol as a standard and 12 DES, of which glycerol:trehalose gave the best results. Pre-incubation for 30 minutes caused better growth after thawing than longer preincubation (1 hour, 3 hours). Gradual freezing, after 1 hour pre-incubation, had no significant effect on the success of cryopreservation. Although in all experiments with the use of glycerol and glycerol:trehalose DES as cryoprotectants, the survival of the water mould S. parasitica after thawing was 100%, a longer freezing time (32 days versus 7 days) caused a slower regeneration of the mycelium. In conclusion, further optimisation of the cryopreservation protocol of the water mould S. parasitica is needed.

• Keywords: Saprolegnia parasitica; vodena plijesan; krioprotektanti; niskotemperaturna eutektička otapala
• Keywords (english): Saprolegnia parasitica; water mould; cryoprotectants; deep eutectic solvents
• Language: croatian
• URN:NBN: urn:nbn:hr:159:254716
• Study programme: Title: Biotechnology; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-11-11 13:24:53