Nastja Novak (Avtor), Petra Kocbek (Mentor), Aleš Lapanje (Komentor)

Povzetek

Ustna votlina predstavlja habitat številnim vrstam mikroorganizmov, ki v določenih pogojih lahko povzročijo različne bolezni, kot je parodontalna bolezen. Ker trenutne smernice zdravljenja ne vodijo do želenih rezultatov, so aktualni novi pristopi zdravljenja, med katere spada tudi lokalno zdravljenje s probiotiki. To zdravljenje temelji na inhibiciji rasti parodontalno patogenih bakterij in zaradi razrasti probiotičnih bakterij ter nastanka probiotičnega biofilma onemogoči ponovno kolonizacijo patogenih mikrobov. Namen magistrske naloge je bil pridobiti avtohtone bakterijske seve s protimikrobnim učinkom proti parodontalno patogeni bakteriji Aggregatibacter actinomycetemcomitans iz ustne votline posameznikov z majhnim številom zalivk na zobeh. Za pridobitev bakterijskih sevov s protimikrobnim učinkom smo razvili protokol vzorčenja ustne votline. Ovrednotili smo vpliv vnosa hrane na pojavnost sevov s protimikrobnim učinkom, zato smo vzorčenje izvedli v različnih časovnih točkah (jutranje, popoldansko vzorčenje in t.i. vzorčenje z minimalnim vplivom hrane). Preverili smo tudi ali so sevi s protimikrobnim učinkom prisotni v vseh ekoloških biotopih, zato smo vzorčili štiri različna mesta v ustni votlini (slino, bukalno sluznico, jezik in površino zob). Uporabili smo obogatitvena gojišča za izolacijo aktinomicet in po Gramu pozitivnih bakterij. V naslednjem koraku smo razvili metodo za vrednotenje protimikrobnega učinka izoliranih bakterij na bakteriji Escherichia coli in Aggregatibacter actinomycetemcomitans. Ugotovili smo, da izolirani bakterijski sevi iz ustne votline kažejo protimikrobni učinek na A. actinomycetemcomitans iz vseh mest ustne votline in iz vseh časovnih točk. S tem smo pokazali, da prispeva k zdravi ustni flori tako hipoteza o ekološki niši kot hipoteza emigracije in imigracije. V zaključni fazi raziskave smo izolirane bakterije s protimikrobnim učinkom karakterizirali z molekularnimi mikrobiološkimi metodami. Večina sevov s protimikrobnim učinkom izoliranih iz ustne votline, ki delujejo proti A. actinomycetemcomitans spada v rod Bacillus. Na podlagi pridobljenih rezultatov lahko predlagamo bakterije rodu Bacillus kot probiotični način zdravljenja parodontalne bolezni. Naša študija je prvi primer, kjer smo iz ustne votline izolirali bakterije rodu Bacillus s protimikrobnim učinkom na parodontalno patogeno bakterijo A. actinomycetemcomitans.

Ključne besede

Aggregatibacter actinomycetencomitans;ustna votlina;gojitvene metode;protimikrobne snovi;Bacillus;

Podatki

Jezik: Slovenski jezik
Leto izida:
Tipologija: 2.09 - Magistrsko delo
Organizacija: UL FFA - Fakulteta za farmacijo
Založnik: [N. Novak]
UDK: 616.311.2:615(043.3)
COBISS: 4326769 Povezava se bo odprla v novem oknu
Št. ogledov: 293
Št. prenosov: 65
Ocena: 0 (0 glasov)
Metapodatki: JSON JSON-RDF JSON-LD TURTLE N-TRIPLES XML RDFA MICRODATA DC-XML DC-RDF RDF

Ostali podatki

Sekundarni jezik: Angleški jezik
Sekundarni naslov: Isolation and characterization of bacteria with antimicrobial effect against bacteria Aggregatibacter actinomycetencomitans
Sekundarni povzetek: The oral cavity represents a habitat for many species of microorganisms, which in certain conditions can cause various diseases, such as periodontal disease. Since current treatment guidelines do not lead to desired results, new approaches are under development, which also includes local treatment with probiotics. This approach is based on the inhibition of the growth of periodontal pathogenic bacteria, and due to the growth of probiotic bacteria and the formation of a probiotic biofilm prevents re-colonization of pathogenic microbes. The purpose of the master's thesis was to obtain autohtonous bacterial strains with antimicrobial effect against periodontal pathogenic bacteria Aggregatibacter actinomycetemcomitans from the oral cavity of individuals with small number of dental fillings. In order to obtain bacterial strains with antimicrobial effect, we have developed a protocol of sampling the oral cavity. We evaluated the effect of food intake on the appearance of strains with antimicrobial effects, so we performed the sampling at different time points (morning, afternoon, and a so-called sampling with minimal impact of food). We also checked whether the strains with antimicrobial activity are present in all ecological biotopes, therefore samples were taken from four different positions in the mouth (saliva, buccal mucosa, tongue and the surface of the tooth). We used the enrichment medium for the isolation of actinomycetes and Gram-positive bacteria. In the next step, we have developed a method for the evaluation of the antimicrobial effect of the isolated bacteria on the bacteria Escherichia coli and Aggregatibacter actinomycetemcomitans. We have found that isolated bacterial strains from the oral cavity show an antimicrobial effect on A. actinomycetemcomitans from all sites of the oral cavity and from all time points. Thus, we have shown that the hypothesis of ecological niche, and the hypothesis of emigration and immigration contribute to a healthy oral flora. In the final phase of the research, we characterized isolated bacteria with antimicrobial effect by molecular microbiological methods. Most strains isolated from the oral cavity with antimicrobial effect against A. actinomycetemcomitans belong to the genus Bacillus. Based on the obtained results we may suggest the genus Bacillus as a probiotic method for treatment of periodontal disease. Our study is the first case in which we have isolated bacteria from the oral cavity belonging to the genus Bacillus, with antimicrobial effect on periodontal pathogenic bacteria A. actinomycetemcomitans.
Sekundarne ključne besede: Parodontalne bolezni;
Vrsta dela (COBISS): Magistrsko delo/naloga
Komentar na gradivo: Univ. Ljubljana, Fak. za farmacijo
Strani: VII, 75 f.
ID: 12044627