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  • Author: Joanna Pławińska-Czarnak x
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Open access

Magdalena Kizerwetter-Świda, Dorota Chrobak-Chmiel, Magdalena Rzewuska, Joanna Pławińska-Czarnak and Marian Binek

Abstract

Introduction: Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) belonging to the clonal complex 398 (CC398) emerged recently in livestock as a new type of MRSA, which may cause zoonotic infections. This study presents data on the characterisation of S. aureus isolated from the meat processing plants. Material and Methods: S. aureus was isolated from 90 samples collected in the raw meat warehouse, from devices and surfaces of meat processing plants, and from finished meat products. The isolates were subjected to molecular analysis in order to investigate the presence of enterotoxin genes, the mecA gene, and to verify whether they belong to the clonal complex 398. The genetic relatedness of the isolates was determined using pulsed-field electrophoresis. Likewise, antimicrobial susceptibility was tested. Results: From 21 S. aureus strains isolated, five belonged to the CC398, two of which were recognised as MRSA and three as methicillin-sensitive Staphylococcus aureus (MSSA). The most prevalent enterotoxin genes were seg and sei. Two MRSA CC398 isolates, three MSSA CC398, and one MSSA were classified as multidrug-resistant. Conclusion: The first isolation of MSSA CC398 from beef in Poland indicates contamination of beef by strains belonging to this clonal complex. The occurrence of multidrug-resistant enterotoxigenic S. aureus isolates in the finished meat products constitutes a potential risk for the consumers.

Open access

Joanna Pławińska-Czarnak, Joanna Zarzyńska, Janusz Bogdan, Alicja Majewska, Marek Karwański, Magdalena Kizerwetter-Świda, Jarosław Kaba, Krzysztof Anusz and Emilia Bagnicka

Abstract

The goat (Capra hircus) is a perfect animal model for analyzing the transcriptome of milk somatic cells (MSCs), as sufficient numbers of somatic cells in goat milk, i.e., exfoliated epithelial cells, can be obtained using noninvasive methods. RNA integrity and purity are the first and most important parameters qualifying samples for transcriptomic tests and next-generation sequencing, as RNA quality influences experimental results. The aim of this study was to optimize a method for obtaining high-quality RNA from goat MSCs, irrespective of effects like breed, lactation stage, health status (e.g., with or without small ruminant lentivirus [SRLV] infection), or number of somatic cells. Milk samples were obtained from goats of two Polish breeds in various lactation stages and in different parities, and from goats infected and not infected with SRLV. Altogether, 412 MSC samples were examined: 206 using method A with fenozol and 206 using method B with QIAzol. Though the overall purity (measured as absorbance ratios at 260 nm/280 nm and 260 nm/230 nm) of the RNA material was comparable, the average yield of RNA isolated using method A was 11.9 µg, while method B’s average yield was 29.9 µg. Moreover, method B resulted good quality RNA suitable for transcriptome analysis. Results were confirmed by RT-qPCR, using 18S rRNA and RPLP0 as the reference genes. The application of our modified treatment method was successful in obtaining high-integrity samples for transcriptomic or next-generation sequencing analysis. Using a 400 mL milk sample cooled in ice directly after milking, securing the cooling chain process from milking to MSC isolation, and applying method B to isolate RNA, we obtained good RNA quality irrespective of the goats’ breed, lactation stage, parity, milk yield, SRLV infection, and even milk yield and number of somatic cells in milk.