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Sinhronizacija bakterijske kulture Escherichia coli in njen vpliv na namnoževanje faga T4
ID Hrovat, Jan (Author), ID Podgornik, Aleš (Mentor) More about this mentor... This link opens in a new window

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Abstract
Bakteriofagi predstavljajo obetavno alternativo antibiotikom, njihova učinkovita proizvodnja pa je odvisna od fiziološkega stanja gostiteljskih bakterij. Namen magistrske naloge je bil vzpostaviti vsaj delno sinhronizacijo celične delitve v kulturi Escherichia coli z metodo cikličnega dohranjevanja, ter ovrednotiti vpliv faze sinhronizacijskega cikla na učinkovitost namnoževanja bakteriofagov T4. Sinhronizacijo smo izvedli v s kemijsko definiranim gojiščem, pri čemer smo po vsaki podvojitvi bakterijske populacije odstranili polovico gojitvene brozge in jo nadomestili s svežim gojiščem. Potek procesa smo spremljali z meritvami raztopljenega kisika, optične gostote in številčnostjo živih bakterij (CFU). Stopnjo sinhronizacije smo ovrednotili s sinhronizacijskim indeksom Blumenthala in Zahlerja, ki smo ga izračunali na podlagi eksperimentalno pridobljenih vrednosti CFU. Za določitev vpliva fiziološkega stanja bakterij na razmnoževanje fagov smo v različnih časovnih točkah sinhronizacijskega cikla izvedli infekcijske poskuse z bakteriofagom T4. Ciklično gojenje je omogočilo ponovljive rastne cikle, pri katerih je bila večina celičnih delitev skoncentrirana v drugi polovici cikla, kar potrjuje doseženo delno sinhronizacijo bakterijske populacije. Dokazali smo tudi, da sinhronizacija bakterijske kulture vpliva na uspešnost namnoževanja fagov. V primeru infekcije v optimalnem trenutku sinhronizacijskega cikla se adsorpcijska konstanta ne spremeni, brstno število se poveča, latentna perioda pa se skrajša. Dobljeni rezultati predstavljajo osnovo za optimizacijo proizvodnje bakteriofagov.

Language:Slovenian
Keywords:sinhronizacija, cikel, bakteriofagi, bakterijska kultura, indeks sinhronizacije, metabolna aktivnost, brstno število, latentna perioda, adsorpcijska konstanta
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Year:2026
PID:20.500.12556/RUL-188212 This link opens in a new window
COBISS.SI-ID:292050691 This link opens in a new window
Publication date in RUL:19.09.2026
Views:104
Downloads:24
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Secondary language

Language:English
Title:Synchronization of Escherichia coli bacterial culture and its impact on T4 phage propagation
Abstract:
Bacteriophages represent a promising alternative to antibiotics, however, their efficient production depends on the physiological state of the host bacteria. This master's thesis aimed to establish at least partial synchronization of an Escherichia coli culture using the cyclic fed-batch technique, and to evaluate the influence of the synchronization cycle phase on the reproduction efficiency of bacteriophages T4. The synchronization was carried out in a chemically defined medium in which, after each doubling of the bacterial population, half of the culture broth was removed and replaced with a fresh medium. The progress of the process was monitored by measuring dissolved oxygen, optical density, and the number of viable bacteria (CFU). The degree of synchronization was evaluated using Blumenthal’s and Zahler’s synchronization index, which was calculated based on the experimentally obtained CFU values. To determine the influence of the physiological state of the bacteria on phage reproduction, infection experiments with bacteriophage T4 were performed at various time points of the synchronization cycle. Cyclic cultivation enabled repeatable growth cycles, in which most cell divisions were concentrated in the second half of the cycle, confirming the achieved partial synchronization of the bacterial population. It has also been proved that synchronization of the bacterial culture has influence on the success of phage reproduction. When infection occurred at the optimal point of the synchronization cycle, the adsorption constant does not change and burst size increased, while the latent period was shortened. The obtained results provide a basis for optimizing bacteriophage production.

Keywords:synchronization, cycle, bacteriophages, bacterial culture, index of synchronization, metabolic activity, burst size, latent period, adsorption constant

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