Virulence and antimicrobial susceptibility profile of Listeria monocytogenes isolated from frozen vegetables available in the Egyptian market

Abstract

Listeria monocytogenes is among the most important foodborne pathogens. It may enter foodprocessing environments through raw materials, handlers or equipment and may persist due to ineffective cleaning or sanitation. The bacterium can be isolated from both frozen vegetables and fresh food substances. This study aimed to estimate the prevalence of L. monocytogenes in spices and frozen vegetables and screen for some virulence factors and drug-resistance determinants of the isolated bacteria. First, conventional microbiological methods were used for the isolation and identification of bacteria. Next, the identity of isolated bacteria was confirmed by molecular techniques, and the virulence genes iap and hlyA were identified by real-time polymerase chain reaction (PCR). The hemolytic activity of the isolates was assessed by cultivation on sheep blood agar. Furthermore, the antimicrobial susceptibility of confirmed L. monocytogenes isolates was tested by the disk diffusion method against 10 antibiotics. Out of 331 vegetable samples, 47 isolates were confirmed to contain L. monocytogenes, whereas none of 40 spice samples tested positive. All isolates were positive for iap and hlyA genes. Susceptibility testing indicated that all isolates were sensitive to trimethoprim/ sulfamethoxazole, but only 36% were sensitive to penicillin G, while 100% and 70% showed intermediate resistance to chloramphenicol and erythromycin, respectively. All tested isolates were resistant to amoxicillin, gentamicin and norfloxacin; on the other hand, 90, 86 and 84% of the tested strains were resistant to ciprofloxacin, ceftazidime/clavulanic acid and amikacin, respectively. In summary, L. monocytogenes isolates disseminated in frozen vegetable samples from the Egyptian market were highly virulent, entirely multiple-drug resistant and were enriched in iron-containing vegetables. Since L. monocytogenes is primarily pathogenic to humans and causes a life-threatening disease, there is a potential infection risk for people who usually deal with frozen vegetables before cooking. Hence, surveillance to L. monocytogenes in frozen products, together with implementation of tight measures would be valuable in preventing listeriosis, and are highly recommended