Osamitev in karakterizacija bakterije Clostridium difficile iz prsti z domačih vrtov

Smrke, Julija (author), Ambrožič Avguštin, Jerneja (mentor), Janežič, Sandra (co-mentor)

Clostridium difficile predstavlja enega najpomembnejših povzročiteljev črevesnih okužb, povezanih z zdravstvom, vse pogosteje pa je tudi vzrok driske v domačem okolju. Okolje domnevno predstavlja enega od potencialnih virov okužbe. Clostridium difficile je namreč prisotna v prsti, vodi, hrani in pri živalih. V našem delu smo se osredotočili na osamitev in karakterizacijo bakterije C. difficile iz prsti z domačih vrtov, natančneje na prst iz kompostnikov in zelenjavnih ter cvetličnih vrtov. Na 5 različnih lokacijah smo skupaj nabrali 25 vozorcev prsti. Prisotnost C. difficile smo dokazali na 4 od 5 lokacijah in v 12 od 25 vzorcev (48 %). Ugotovili smo, da je pogostnost in raznolikost PCR-ribotipov večja v prsti kompostnikov, v primerjavi s prstjo iz vrtov. Razlog temu smo pripisali življenjskim pogojem, naklonjenim anaerobnim organizmom, in dejstvu, da na kompostnike odlagamo različne organske odpadke, ki so lahko kontaminirani s sporami C. difficile. Skupno smo iz 12 pozitivnih vzorcev osamili 132 izolatov C. difficile, ki smo jih z metodo PCR-ribotipizacije (metodo izbora za genotipizacijo bakterije C. difficile) uvrstlili v 10 PCR-ribotipov, med katerimi sta prevladovala PCR-ribotipa SLO 239 (26,5 %) in 001/072 (20,5 %). Skupno so bili 4 od 10 PCR-ribotipov toksigeni, ti so 001/072, 005 in 081 (vsi trije toksinotip 0) in 023 (toksinotip IV). Nekateri genotipi, izolirani iz prsti so enaki kot tisti, ki se pojavljajo tudi pri ljudeh in živalih v Sloveniji. Naši rezultati kažejo, da okolje lahko predstavlja potencialni vir okužbe z bakterijo C. difficile.

• Language: Slovenian
• Keywords: Clostridium difficile, prst, kompost, PCR-ribotip
• Work type: Master's thesis/paper
• Organization: BF - Biotechnical Faculty
• Year: 2017
• PID: 20.500.12556/RUL-92316
• COBISS.SI-ID: 4363599
• Publication date in RUL: 18.05.2017

Secondary language

• Language: English
• Title: Isolation and characterization of Clostridium difficile from soil from domestic gardens

Abstract

Clostridium difficile is one of the most important causes of intestinal infections associated with health care institutions. In recent years it has also become recognized as an important cause of diarrhea in community. Clostridium difficile has been isolated from soil, water, food, and from the feces of different animals. Different environments could therefore represent potential sources of infections. The aim of our study was to isolate and characterize C. difficile from soil samples from domestic gardens, more accurately from the soil of composters, and from vegetable and flower gardens. Sampling was performed at five different locations. At each location 5 soil samples were collected. C lostridium difficile was present in 12 out of 25 (48 %) samples collected on 4 different locations. Prevalence and diversity of PCR-ribotypes in the soil samples collected from composters were greater compared to soil from gardens (vegetables or flower). The reason may be environmental conditions which are in favor of anaerobic organisms and the fact, that mostly organic waste is being deposited on composters which might contaminate them with C. difficile spores. In total, we isolated 132 isolates of C. difficile. All isolates were distributed into 10 different PCR ribotypes (PCR ribotyping is a method of choice for genotyping of C. difficile), among which the most common were PCR ribotypes SLO 239 (26,5%) and 001/072 (20,5%). Four out of 10 PCR ribotypes were toxigenic, i.e. 001/072, 005 and 081 (all of them toxinotype 0) and 023 (toxinotype IV). Some of the genotypes isolated from the soil were the same as those occurring in humans and animals in Slovenia. Our results indicate that domestic environment could represent a potential source of infection with C. difficile.

• Keywords: Clostridium difficile, soil, compost, PCR-ribotype