Koncept kontrole Listeria monocytogenes u pogonu za preradu mesa uz upotrebu sekvenciranja kompletnog genoma u cilju postizanja bezbednosti hrane

Abstract

L.monocytogenes, fakultativni intracelularni patogen, je široko rasprostranjena u razliĉitoj sveţoj i preraĊenoj hrani. Ovaj patogen je uzroĉnik listerioze, retkog ali ozbiljnog oboljenja ljudi i ţivotinja, izazvanog konzumacijom kontaminirane hrane. L.monocytogenes moţe da preţivi i raste u širokom rasponu razliĉitih uslova sredine u kojoj se nalazi, bilo da je ona prirodna ili sredina modifikovana aktivnošću ĉoveka. Usled sposobnosti da preţivi i raste na niskim temperaturama, u sredinama sa visokom koncentracijom soli, smanjenom pH i niskim vrednostima aktivnosti vode, ĉest je stanovnik pogona za proizvodnju hrane. Zbog svoje sveprisutnosti u ţivotnoj sredini, kontrola pojave L.monocytogenes u pogonima za proizvodnju hrane predstavlja ozbiljan izazov za sistem bezbednosti hrane. U cilju unapreĊenja sistema bezbednosti hrane, uporedo sa razvojem legislative i standarda, razvijaju se i nove metode i tehnike identifikacije mikroorganizama. Sekvenciranje kompletnog genoma (eng. Whole genome sequencing - WGS) predstavlja ultimativnu metodu za ispitivanje prisustva patogena i otkrivanje njihovog porekla u hrani. Delikates proizvodi od mesa spremni za jelo, identifikovani su kao jedan od ĉestih izvora epidemija listerioze. Prisustvo L.monocyogenes u pogonu za proizvodnju tradicionalnih proizvoda od mesa u Crnoj Gori, ispitivano je tokom perioda od 2011. do 2014. godine. Za posmatrani period izolovano je 20 izolata L.monocytogenes iz proizvoda od mesa i pogona za njihovu proizvodnju koji su podvrgnuti molekularnoj karakterizaciji, serotipizaciji uz pomoć multipleks PCR-a i sekvencera nove generacije (eng. Next generation sequencing – NGS), utvrĊivanju potencijalne antimikrobne rezistencije na osnovu NGS podataka o prisustvu secifiĉnih gena rezistencije i skriningu na prisustvo odreĊenih markera rezistencije na dezinficijense...

L. monocytogenes, facultative intracellular pathogen, can be found in different types of raw and processed foods. This pathogen causes listeriosis, a rare but severe disease in humans and animals, which is spread primarily through the consumption of contaminated food. L. monocytogenes can survive and grow in a wide spectrum of environmental conditions, weather natural or habitats created by the human activity. Due to its ability to survive low temperatures, higher sodium nitrate concentrations, low pH and water activity, L. monocytogenes can frequently be found in the food processing environment. The ubiquitous nature of L. monocytogenes poses a significant encounter to its control in the food production environment, representing a great challenge for food safety. With the aim of improvement of the efficiency of the food safety systems, in parallel to the development of legislation and standards, the new methods and techniques for identification of microorganisms have been devised. Whole genome sequencing (WGS) technique represents the ultimate method for pathogen detection and for determination of their origin in food products. Ready-To-Eat (RTE) delicatessen meat products have repeatedly been identified as a source of listeriosis outbreaks. The presence of Listeria monocytogenes was evaluated in the meat processing facility in Montenegro in the period from 2011 to 2014. In total, 20 isolates of L. monocytogenes, isolated from traditional meat products and environmental swabs, were subjected to molecular characterization, serotyping by both multiplex PCR and next generation sequencing (NGS), testing to their potential antimicrobial resistance (AMR) by extraction of specific genes from NGS data and screening for the presence of some disinfectant resistance markers...