Staphylococcal food poisoning (SFP) is an important foodborne disease worldwide, and milk and milk products are commonly associated with SFP outbreaks. The objectives of this study were to investigate the distribution of staphylococcal enterotoxin (se) genes in Staphylococcus aureus from raw cow's milk and milk products and to assess their genetic background with the spa typing method. Of the 549 samples (297 bulk milk and 162 milk product samples) collected from Tigray region, Northern Ethiopia, 160 (29.1%) were positive for S. aureus, of which 82 (51%) were found to harbor se genes by a modified multiplex PCR. Nine se genes were identified: sea (n = 12), seb (n = 3), sec (n = 3), sed(n = 4), seg (n = 49), seh (n = 2), sei (n = 40), sej (n = 1), and tsst-1 (n = 24). The classical type of genes accounted for 27%. Of the 82 enterotoxigenic isolates, 41.5 and 12.4% harbored two or more se genes, respectively. The highest gene association was observed between sei and seg, whereas sea and seb were always found together with the new types of se genes. Altogether, 18 genotypes of toxin genes were identified, and 33% of the samples contained > 5 log CFU ml(-1) S. aureus. spa typing identified 22 spa types and three novel spa sequences, which showed the high genetic diversity of the isolates. No apparent relationship was observed between spa type and se genes. Of the 25 spa types, 13 (52%) were from raw milk, 3 (12%) from milk products, and 9 (36%) from both types of sample. Types t314 (20.7%,n = 17), t458 (18.3%, n = 15), and t6218 (9.8%, n= 8) were the most common spa types identified and were widely distributed in three of the eight studied localities. This is the first study from the Tigray region to report the high distribution of enterotoxigenic S. aureus with a diversified genetic background from dairy food. The study may provide valuable data for microbial food safety risk assessment, molecular epidemiology, and phylogenetic studies of S. aureus in Ethiopia.
Staphylococcus aureus is an important pathogen that can cause Staphylococcal Food Poisoning (SFP).Milk and dairy products are frequently contaminated by this bacterium. In this study, 310 samples (168 bovine raw milk and 142 dairy products) were collected in the Tigray region of Northern Ethiopia, with the objective of detection and enumeration of S. aureus and other Staphylococcus species. Baird-Parker agar for isolation and 16S rRNA gene sequencing for species identification were employed. S. aureus was found in 38.7% of samples with mean count of 4.35 +/-0.97 log 10 CFU ml -1 . The prevalence of S. aureus was significantly greater in raw milk samples (47%) than in dairy products (28.8%). Of all S. aureus positive samples, 34.2% contained >5 log 10 CFU ml -1 . Samples from cafeterias and restaurants showed the greatest prevalence of S. aureus (P<0.05) compared to other sampling points. Samples from small-scale dairies were twice more likely to be contaminated by S. aureus than traditionally managed dairies (P<0.05, OR=2.0). Coagulase-negative staphylococci (CNS) were found in 51.6% of the samples, with mean count of 6.0±1.21 log 10 CFU ml -1 . Ten species of CNS were identified and S. epidermidis (36.13%) was the most frequent. The frequency of isolation found in this study indicates that S. aureus and other Staphylococcus spp. may impose public health hazard in dairy products. Therefore, further studies on the enterotoxigenic potential of the isolates, and molecular epidemiology to trace the sources of the contamination are recommended.
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