Biogenic amine content during the production and ripening of Sremski kulen, Serbian traditional dry fermented sausage.

Sremski kulen is a wide diameter dry fermented sausage, produced from pork, seasoned with red spicy paprika, stuffed into pork cecum, and preserved by smoking, fermentation and drying. Due to specific ripening process, Sremski kulen is suitable for the accumulation of biogenic amines. Therefore, the aminogenesis was studied in traditionally produced Sremski kulen, taking into account the physicochemical parameters and microbial counts. The content of six biogenic amines (tryptamine, phenylethylamine, tyramine, histamine, putrescine, and cadaverine) was analyzed using high-performance liquid chromatography. The ripening process of Sremski kulen was slow followed by changes in aw and pH value as well as expressed proteolysis. The autochthonous microbiota showed pronounced decarboxylase activity. Tryptamine and phenylethylamine were detected at each examined ripening stage while histamine was not detected until the end of ripening (16.55 ± 2.33 mg/kg). Tyramine, cadaverine, and putrescine content significantly increased during the ripening period (p < .05). In the final product, cadaverine was the dominant biogenic amine (132.40 ± 5.05 mg/kg), followed by tyramine (115.80 ± 15.46 mg/kg) and putrescine (68.55 ± 2.39 mg/kg). Although the long ripening period greatly contributed to the accumulation of biogenic amines in final product, their content are not of concern from product safety aspects, but requires improvement in hygiene of production process.

Seroprevalence of hepatitis E in pigs and wild boars in the region of the city Belgrade.

INTRODUCTION: Hepatitis E is considered an emerging human viral disease with many evidences of zoonotic nature of disease, and swine are the main reservoir of HEV. The aim of this study was to determine HEV seroprevalence in commercial pig farms, backyard pigs, slaughtered pigs and wild boars in the region of the city Belgrade. METHODOLOGY: A total of 405 sera samples: 150 samples from 3 commercial pig farms, 70 samples from backyard pigs, 119 samples from slaughtered pigs and 66 samples from wild boars of the region of the city Belgrade, Serbia were analysed by commercial ELISA test. RESULTS: The overall HEV seroprevalence in 3 commercial pig farms was 55.33% (83/150). All tested farms (farm A, B and C) were positive on the presence of anti-HEV antibodies, respectively 58% (29/50), 54% (27/50) and 54% (27/50). From 70 tested backyard pigs, 75.71% (53/70) were tested seropositive. In total, 26 backyard pig holidngs were confirmed as positive to anti-HEV antibodies (81.25%). At slaughterhouse, 25% (8/32) weaned piglets and 20.69% (18/87) fattening pigs were tested positive on anti-HEV antibodies. Overall HEV seroprevalence in tested wild boar population was 52.25% (36/66). CONCLUSIONS: Detected very high seroprevalence of anti-HEV antibodies indicated an active circulation of HEV, being enzootic in the swine population, and wild boars, as well, in the region of the city Belgrade.

Screening and Molecular Characterization of Hepatitis E Virus in Slaughter Pigs in Serbia.

Hepatitis E virus (HEV) is a zoonotic virus that can cause acute hepatitis in humans. Besides the fecal-oral route, transmission can occur by consumption of undercooked pig liver. Genotype 3 is the most frequent genotype found in Europe. Studies on HEV in slaughter-age pigs have not been conducted in Serbia so far. Pork meat production and consumption in Serbia is on average, higher than in the rest of Europe. With the aim to identify the circulating HEV genotypes, pig livers and swab samples from three pig slaughterhouses located in three different sub-regions of Serbia were collected. A nested RT-PCR was used to amplify the hypervariable HEV ORF-1 region (334 bp). The amplicons yielded in this study were sequenced, and a molecular phylogeny analysis based on the maximum likelihood method, including HEV sequences reported in several other countries, was performed. The average prevalence of HEV genotype 3 in 3-month-old pigs was 34%. Phylogenetic analysis revealed the majority of HEV amplification fragments from Serbia were grouped in four clades within sub-genotype 3a and were also genetically related to German, Italian, Slovenian, and American HEV sequences. Sub-genotypes 3b and 3j were also found in a single pig each. This study provides the first analysis of the genetic diversity and circulation dynamics of HEV in pigs at slaughterhouses in Serbia.

Prevalence of Salmonella enterica in slaughtered pigs in Serbia: Serotyping, PFGE-genotyping and antimicrobial resistance.

INTRODUCTION: The aim of this study was to determine the prevalence of Salmonella along the slaughter line and to identify possible critical control points in one slaughterhouse facility located in the city of Belgrade. METHODOLOGY: In total, 700 samples were tested: two swabs from both sides of carcass were taken from each of 100 pigs. In this way, 200 pig skin swab samples were taken after stunning, 200 after processing and 200 after chilling. Additional 100 samples of ileal contents were also taken from the same pigs to obtain a collection of 270 isolates. All samples were analyzed using standard culture methods and serotyping.  PFGE was performed for 27 isolates. Determination of antimicrobial resistance was performed by E-test. RESULTS: In total, 47 (23.5%) swab samples were positive for the presence of Salmonella after stunning. After processing, Salmonella was isolated in two swab samples (1%), whereas all samples which were collected after chilling were negative for Salmonella. The sampling of ileal contents was positive for five Salmonella isolates (5%). The most frequently isolated serotypes were S. Derby (90.74%), S. Infantis (5.56%) and S. Typhimurium (3.7%). All tested isolates were resistant to tetracycline. Resistance was recorded to nalidixic acid (23.3%), ciprofloxacin (20%), ampicillin (10%) and chloramphenicol (14.4%), as well. The PFGE results indicated that isolates had a high genetic similarity. CONCLUSIONS: The investigation has confirmed that bacteriological examinations of carcass swabs, as well as ileal content, could be used to assess the carriage of salmonellae in pigs at the time of slaughter.