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Odpornost mlečnokislinskih bakterij in bifidobakterij iz humanega kolostruma in mleka proti tetraciklinu in eritromicinu
Pukmajster, Janja (Author), Bogovič Matijašić, Bojana (Mentor) More about this mentor... This link opens in a new window, Mohar Lorbeg, Petra (Co-mentor)

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Abstract
Namen magistrske naloge je bil raziskati, ali mlečnokislinske bakterije, predvsem laktobacili, in bifidobakterije iz humanega mleka, kolostruma in zrelega mleka, vsebujejo gene za odpornost proti tetraciklinu in eritromicinu, kar predstavlja določeno tveganje za prenos odpornosti na patogene bakterije v dojenčkovem črevesju. Odpornost proti kanamicinu smo priključili iz razloga, ker je bila v predhodnih raziskavah teh sevov nepričakovano pogosta. Tako smo raziskali, ali so v njihovem genomu prisotni pridobljeni geni z zapisi za odpornost proti izbranim antibiotikom. En vzorec kolostruma in 38 vzorcev mleka smo nacepili na gojišči Rogosa in WCA-mup ter osamili 243 izolatov. Rezultati sekvenciranja izolirane DNA so pokazali, da smo s teh gojišč osamili bakterije iz rodov Lactobacillus, Propionibacterium, Streptococcus, Bifidobacterium in Actinomyces. Tem izolatom smo priključili še 56 predhodno izoliranih bakterijskih sevov iz rodov Lactobacillus in Bifidobacterium. Fenotipsko odpornost smo ugotavljali s pomočjo mikrodilucije. Pri sevih, ki so izkazovali fenotipsko odpornost proti določenemu antibiotiku, smo z uporabo PCR metode preverjali prisotnost najbolj znanih genov, povezanih z odpornostjo proti temu antibiotiku. Fenotipsko odpornost proti eritromicinu smo zaznali pri 3 sevih, proti tetraciklinu pri 10 sevih in proti kanamicinu pri 52 sevih, medtem ko prisotnosti genov erm(A), erm(B), erm(C), erm(T), tet(A), tet(B), tet(C), tet(M), tet(O), tet(W), aac(6’)-aph(2’’), ant(6)-Ia in aph(3’)-IIIa nismo potrdili pri nobenem od teh sevov.

Language:Slovenian
Keywords:humano mleko, mlečnokislinske bakterije, bifidobakterije, odpornost proti antibiotikom, eritromicin, tetraciklin, geni za odpornost
Work type:Master's thesis/paper (mb22)
Tipology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Year:2017
Publisher:[J. Pukmajster]
UDC:579.24/.26:612.664:615.33
COBISS.SI-ID:4858488 Link is opened in a new window
Views:467
Downloads:313
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Secondary language

Language:English
Title:The resistance of lactic acid bacteria and bifidobacteria from human colostrum and milk to tetracycline and erythromycin
Abstract:
The aim of this master's thesis was to investigate whether lactic acid bacteria, especially lactobacilli, and bifidobacteria from human milk, colostrum and mature milk, contain tetracycline and erythromycin resistance genes, which presents a certain risk of spread of antibiotic resistance to pathogenic bacteria in the infant's gut. Kanamycin was subsequently added to the study for a reason that in previous studies the resistance of lactobacili to this antibiotic was unexpectedly frequent. Thus, we investigated whether the genes associated with the resistance to selected antibiotics are present in the genome of lactic acid bacteria and bifidobacteria. One sample of colostrum and 38 samples of milk were plated on Rogosa and WCA-mup medium. 243 bacterial strains were isolated. The results of the sequencing of isolated DNA showed that bacteria belonged to the genera Lactobacillus, Propionibacterium, Streptococcus, Bifidobacterium and Actinomyces. We also included 56 pre-isolated bacterial strains from the genera Lactobacillus and Bifidobacterium. Phenotypic resistance was determined by microdilution. In phenotypic resistant strains we investigated the presence of the most well-known genes associated with resistance to a particular antibiotic with PCR. Phenotypic resistance to erythromycin was detected in 3 strains, tetracycline in 10 strains, and kanamycin in 52 strains, while the presence of investigated genes erm(A), erm(B), erm(C), erm(T), tet(A), tet(B), tet(C), tet(M), tet(O), tet(W), aac(6')-aph(2''), ant(6)-Ia and aph(3')-IIIa has not been confirmed in any of the strains.

Keywords:human milk, lactic acid bacteria, Bifidobacterium, antibiotic resistance, erythromycin, tetracycline, antibiotic resistance genes

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