Analiza rizika prisustva Campylobacter jejuni u lancu proizvodnje živinskog mesa

Abstract

Kampilobakterioza je infektivno obljenje ljudi koje uzrokuju termotolerantne vrste roda Campylobacter, prevashodno C. jejuni i C. coli. U savremenom svetu kampilobakterioza je vodeći uzročnik gastroenteritisa sa preko 200 000 obolelih na godišnjem nivou samo u Evropskoj uniji. Epidemiologija uzročnika nije u potpunosti razjašnjena, ali mnogobrojna istraživanja ukazuju na pileće meso kao jedan od glavnih izvora infekcije. U ovom istraživanju ispitano je prisustvo Campylobacter vrsta na teritoriji Republike Srbije, na farmama i klanicama živine, i izvršena tipizacija dobijenih izolata. Pregledano je 233 uzoraka poreklom od živine, od čega 190 sa klanica i 43 sa farmi. Ukupno je prikupljeno 48 Campylobacter spp. izolata, i to 43 sa klanica i 5 sa farmi. Takođe je ispitano 7 izolata poreklom od ljudi od kojih je 5 potvrđeno kao Campylobacter spp., a tipizirana su i 2 referentna soja C. jejuni ATCC 29428 i ATCC 33291. Izolati su potvrđeni do nivoa vrste multiplex-PCR metodom, a tipizacija je izvršena upotrebom Random Amplified Polymorphic DNA (RAPD) i Repetitive Sequence-Based PCR (rep-PCR) metoda uz upotrebu više prajmera. Kao visoko diskriminatorni pokazali su se RAPD prajmeri AP10, AK16 i DJP17, dok su RAPD prajmeri SPH1 i AG15) i rep-PCR prajmer (GTG)5 bili umereno diskriminatorni. Ovi rezultati sugerišu da primena molekularnih metoda RAPD i rep-PCR može biti korisna za tipizaciju izolata Campylobacter vrsta, a dobijeni rezultati se mogu koristiti u epidemiološkim istraživanjima.

Campylobacteriosis is infectious disease of humans caused by thermophilic Campylobacter species, mainly C. jejuni and C. coli. It is a leading cause of human gastroenteritis today with more than 200 000 cases per year in the European Union. The epidemiology of the agent is not completely cleared, but number of investigation indicates that broiler meat plays an important role in human infection. This investigation was performed in order to determine presence of Campylobacter spp. on the broiler farms and slaughterhouses and to establish the genetic profiles and connections beetween the isolates. 233 samples taken at broiler farms (43 samples) and slaughterhouses (190 samples) were investigated, a s well as 7 human isolates and 2 reference strains C. jejuni ATCC 29428 and ATCC 33291. 43 isolates were collected on slaughterhouses and 5 on farms. 5 human isolates were tested. Strain identification was performed by multiplex-PCR analysis. Genotyping was performed by Random Amplified Polymorphic DNA (RAPD) and Repetitive Sequence-Based PCR (rep-PCR) with multiple primers. Best results were obtained with RAPD primers AP10, AK16 and DJP17. RAPD primers SPH1 and AG15, as well as rep-PCR primer (GTG)5 were partly informative and could be used in combination with the previous. There results suggest that molecular techniques RAPD and rep-PCR could be used for genotyping of Campylobacter species and epidemiology of the agent.