Rep-PCR typing of Staphylococcus spp. strains in meat paste production line and identification of their origin

• Main Authors: Ivan Manga, Marcela Klimešová, Jiří Horáček, Ivana Koláčková, Marie Bjelková, Antonín Ponížil, Ludmila Nejeschlebová
• Format: Article
• Language: English
• Published: HACCP Consulting 2015-05-01
• Series: Potravinarstvo
• Subjects: rep-PCR; typing; meat paste; staphylococci; spice
• Online Access: http://www.potravinarstvo.com/journal1/index.php/potravinarstvo/article/view/465

<!--[if gte mso 9]><xml> <o:OfficeDocumentSettings> <o:AllowPNG /> </o:OfficeDocumentSettings> </xml><![endif]--><span style="font-size:10.0pt;font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;mso-fareast-font-family:&quot;Times New Roman&quot;;mso-ansi-language: EN-GB;mso-fareast-language:SK;mso-bidi-language:AR-SA" lang="EN-GB">A meat paste production line and its microbial parameters have been evaluated in single Czech company. The raw meat paste samples before heat treatment were tested positively for the presence of three staphylococci species: <em style="mso-bidi-font-style:normal">Staphylococcus aureus</em>, <em style="mso-bidi-font-style:normal">Staphylococcus haemolyticus</em> and <em style="mso-bidi-font-style:normal">Staphylococcus epidermidis</em>. Subsequent microbial analysis of meat paste components and ingredients (fresh meat, water, spices, equipment) identified only the spices used as positive for <em style="mso-bidi-font-style:normal">S. aureus</em> (coriander, cinnamon, badian, mustard &ndash; (10 - 40 cfu/g)) and <em style="mso-bidi-font-style:normal">S. haemolyticus</em> strains (juniper, </span><span style="font-size: 10.0pt;font-family:&quot;Times New Roman&quot;,&quot;serif&quot;;mso-fareast-font-family:&quot;Times New Roman&quot;; color:black;mso-ansi-language:EN-GB;mso-fareast-language:CS;mso-bidi-language: AR-SA" lang="EN-GB">ginger)</span><span style="font-size:10.0pt;font-family:&quot;Times New Roman&quot;,&quot;serif&quot;; mso-fareast-font-family:&quot;Times New Roman&quot;;mso-ansi-language:EN-GB;mso-fareast-language: SK;mso-bidi-language:AR-SA" lang="EN-GB">. The collection of sixteen collected strains (<em style="mso-bidi-font-style:normal">S. aureus</em> (n = 4), <em style="mso-bidi-font-style: normal">S. haemolyticus</em> (n = 4), <em style="mso-bidi-font-style:normal">S. epidermidis</em> (n = 8)) has been typed with the rep-PCR method utilising (GTG)5 primer. Analysis of the fingerprints using the unweighted pair-group method using arithmetic averages (UPGMA) clustering method revealed presence of eleven strain clusters with similarity lower than 90%: two fingerprint clusters of <em style="mso-bidi-font-style:normal">S. aureus</em>, three individual clusters characteristic for <em style="mso-bidi-font-style:normal">S. haemolyticus</em> and six different <em style="mso-bidi-font-style:normal">S. epidermidis </em>specific clusters. The <em style="mso-bidi-font-style:normal">S. aureus</em> strains from different types of spice were identical, resp. very similar. Molecular tracking composed from the rep-PCR analysis of acquired isolates and comparison among all collected fingerprints confirmed the spices to be the source of both <em style="mso-bidi-font-style:normal">S. aureus</em> and <em style="mso-bidi-font-style: normal">S. haemolyticus</em> strains identified in raw meat paste. The additional rep-PCR analysis of the <em style="mso-bidi-font-style:normal">S. epidermidis</em> collection confirmed usability and performance of this method. The antibiotic susceptibility to fourteen individual antibiotics has been examined among the collected staphylococci strains. The predominant erythromycin resistance (68.8%) was followed with the resistance to amoxicillin/clavulanic acid (56.2%). Other resistances observed were less frequent (clindamycin &ndash; 12.5%, oxacillin &ndash; 6.3%, tetracycline &ndash; 6.3%, sulphamethoxazole-trimethoprim &ndash; 6.3%, chloramphenicol &ndash; 6.3%, novobiocin &ndash; 6.3%). 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