Application of Different Molecular Techniques for Characterization of Catalase‐Positive Cocci Isolated from Sucuk

Abstract

This study was carried out for the characterization and discrimination of the indigenous Gram positive, catalase-positive cocci (GCC) population in sucuk, a traditional Turkish dry-fermented sausage. Sucuk samples, produced by the traditional method without starter culture were collected from 8 local producers in Kayseri/Turkey and a total of 116 GCC isolates were identified by using different molecular techniques. Two different molecular fingerprinting methods; namely, randomly amplified polymorphic DNA-PCR (RAPD-PCR) and repetitive extragenic palindrome-PCR (rep-PCR), were used for the clustering of isolates and identification at species level was carried out by full length sequencing of 16S rDNA. Combining the results obtained from molecular fingerprinting and 16S rDNA sequencing showed that the dominant GCC species isolated from the sucuk samples was Staphylococcus saprophyticus followed by Staphylococcus succinus and Staphylococcus equorum belonging to the Staphylococcus genus. Real-time PCR DNA melting curve analysis and high-resolution melting (HRM) analysis targeting the V1 + V3 regions of 16S rDNA were also applied for the discrimination of isolates belonging to different species. It was observed statistically different Tm values and species-specific HRM profiles for all except 2 species (S. saprophyticus and Staphylococcus xylosus) that have high 16S rDNA sequence similarity. The combination of rep-PCR and/or PCR-RAPD with 16S rRNA gene sequencing was an efficient approach for the characterization and identification of the GCC population in spontaneously fermented sucuk. On the other hand, intercalating dye assays were found to be a simple and very promising technique for the differentiation of the GCC population at species level.

Microbiological changes, shelf life and identification of initial and spoilage microbiota of sea bream fillets stored under various conditions using 16S rRNA gene analysis

“…Macrococcus caseolyticus strain YSY1‐12 found in our study has been reported as part of the spoilage microbiota of water‐boiled salted duck stored at 4 °C . However, M. caseolyticus , previously classified in the genus Staphylococcus ( S. caseolyticus ), has also been isolated from fermented fish products, fermented meat products, fermented sausages, raw milk and cheese …”

Some Chemical Characteristics of Sucuk and Salami Samples Available at Retail in Adana

“…Similarly, Ozturk et al (2014) reported that a w values of 35 sucuk samples obtained from sucuk producers ranged from 0.710 to 0.930. In another study, Kesmen et al (2014) indicated that a w values of sucuk samples collected from 8 local producers in triplicate in Kayseri were between 0.752 and 0.854. Likewise, Table 2 provides a w values of the salami samples.…”

Determination of antibiotic susceptibility and decarboxylase activity of coagulase-negativeStaphylococcusandMacrococcus caseolyticusstrains isolated from fermented Turkish sausage (sucuk)

“…Gram staining, catalase, and coagulase test results showed that among the 67 selected colonies, a total of 61 were Gram‐positive, catalase‐positive, and coagulase negative cocci. In previous studies exhibited that Gram‐positive, catalase‐positive, and coagulase negative cocci are associated with traditional fermented sausages produced in Europe (Bonomo, Ricciardi, Zotta, Sico, & Salzano, ; Cachaldora, Fonseca, Franco, & Carballo, ; Kaban & Kaya, ; Kesmen, Yarimcam, Aslan, & Yetim, ; Landeta et al, ; Marty et al, ; Mauriello, Casaburi, Blainotta, & Villani, ; Resch, Nagel, & Hertel, ; Zdolec et al, ), as confirmed in this study.…”

“…The previous studies showed that S. xylosus, S. equorum , and S. saprophyticus have been the most frequently isolated species from traditional fermented sausages. Other species such as S. carnosus , S. caprae , S. epidermidis , S. hominis , S. succinus , S. sciuri , S. vitulinus , and S. warneri are less frequently identified (Bonomo et al, ; Kaban & Kaya, ; Kesmen et al, ; Leroy, Giammarinaro, Chacornac, Lebert, & Talon, ; Mauriello et al, ). We also isolated 10 Macrococcus caseolyticus (formerly Staphylococcus caseolyticus ) strains from sucuk samples.…”