Salmonella enterica subspecies enterica serovar Typhimurium is the most common zoonotic pathogen in Bulgaria. To allow efficient outbreak investigations and surveillance in the food chain, accurate and discriminatory methods for typing are needed. This study evaluated the use of multiple‐locus variable‐number of tandem repeats analysis (MLVA) and compared results with antimicrobial resistance (AMR) determinations for 100 S. Typhimurium strains isolated in Bulgaria during 2008–2012 (50 veterinary/food and 50 human isolates). Results showed that isolates were divided into 80 and 34 groups using MLVA and AMR, respectively. Simpson's index of diversity was determined to 0.994 ± 0.003 and 0.945 ± 0.012. The most frequently encountered MLVA profiles were 3‐11‐9‐NA‐211 (n = 5); 3‐12‐9‐NA‐211 (n = 3); 3‐12‐11‐21‐311 (n = 3); 3‐17‐10‐NA‐311 (n = 3); 2‐20‐9‐7‐212 (n = 3); and 2‐23‐NA‐NA‐111 (n = 3). No clustering of isolates related to susceptibility/resistance to antimicrobials, source of isolation, or year of isolation was observed. Some MLVA types were found in both human and veterinary/food isolates, indicating a possible route of transmission. A majority (83%) of the isolates were found to be resistant against at least one antimicrobial and 44% against ≥4 antimicrobials. Further studies are needed to verify MLVA usefulness over a longer period of time and with more isolates, including outbreak strains.

(1) Background: HEV is a zoonotic, foodborne pathogen. It is spread worldwide and represents a public health risk. The aim of this study was to evaluate the presence of HEV RNA in farrow-to-finish pig farms in different regions of Bulgaria; (2) Methods: Isolation of HEV RNA from pooled samples of feces was performed using a QIAamp® Viral RNA Mini Kit followed by HEV RNA detection using a single-step real-time RT-PCR with primers and probes targeting the ORF 3 HEV genome; (3) Results: HEV RNA was detected in 12 out of 32 tested farms in Bulgaria (37.5%). The overall percentage of HEV-positive pooled fecal samples was 10.8% (68 of 630 samples). HEV was detected mostly in pooled fecal samples from finisher pigs (66/320, 20.6%) and sporadically from dry sows (1/62, 1.6%) and gilts (1/248, 0.4%); (4) Conclusions: Our results confirm that HEV circulates in farrow-to-finish pig farms in Bulgaria. In our study, we found HEV RNA in pooled fecal samples from fattening pigs (4–6-months age), shortly before their transport to the slaughterhouse indicating a potential risk to public health. The possible circulation of HEV throughout pork production requires monitoring and containment measures.

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This paper presents an outbreak provoked by methicillin-resistant strains of coagulase-positive S. aureus (CPS), produced staphylococcal enterotoxins (se) in pig meatballs and potato salad consumed from 70 people in the village Mamarchevo, Bulgaria. Eighteen women aged 50 to 70 years, and two children aged 4 and 5 years have demonstrated a severe malaise with vomiting and indigestion. Two food samples and isolates of CPS were received in the laboratory of Bulgarian Food Safety Agency. Both samples were found to have a high level of CPS. The level of S. aureus contamination in the potato salad was 8.3 logs CFU/g and 7.7 logs CFU/g in roasted meatballs, which was a significant reason to doubt the production of the toxin. The samples were analyzed according to the European Screening Method v5 using mini VIDAS SET2. The results showed a presence of staphylococcal enterotoxin (TV 2.67 for meatballs and TV 3.27 for potato salad), which was the reason for the ensuing food intoxication. EURL CPS applying quantitative indirect sandwich-type ELISA confirmed the presence of sea, sec and sed in the potato salad and sea and sed in the roasted meatballs. Two CPS isolates were confirmed as S. aureus by a species-specific 23S rRNA targeted PCR test. Real-time PCR method detected sea, sed, seg, sei, sej, and ser genes in S. aureus strains, found in both matrixes. Multiplex PCR method proved the existence of the mecA gene in both S. aureus strains. Resistance to cefoxitin (>16 mg/L), penicillin (>2 mg/L), kanamycin (64 mg/L) and sulfamethoxazole (>512 mg/L) was found.