Microbiological analyses were carried out in duplicate as previously described by Alemán et al. [27 (link)]. Briefly, 10 g of the sample were transferred into sterile bags (Sterilin, Stone, Staffordshire, UK) and mixed with 90 mL of buffered 0.1% peptone water (Oxoid, Basingstoke, UK) in a vertical laminar-flow cabinet (mod. AV 30/70 Telstar, Madrid, Spain). After 1 min shaking in a Stomacher blender (model Colworth 400, Seward, London, UK), appropriate dilutions were prepared for the following determinations: total bacterial counts (TBC) on pour plates of Plate Count Agar (PCA) incubated at 30 °C/72 h; Enterobacteriaceae on double-layered plates of Violet Red Bile Glucose agar (VRBG, Oxoid) incubated at 30 °C/48 h; lactic acid bacteria on double-layered plates of De Man, Rogosa and Sharpe (MRS) agar (Oxoid) incubated at 30 °C/72 h; Staphylococcus coagulase positive (S. aureus and other species) on Baird Parker-RPF agar supplemented with rabbit plasma and fibrinogen (BioMérieux S.A., Marcy l’Etoile, France). Microbiological counts were expressed as the log of the colony-forming units per gram (Log cfu/g) of sample.
Buffered 0.1 peptone water
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
Buffered 0.1% peptone water is a sterile, ready-to-use liquid medium suitable for the preparation of microbial samples. It is formulated to maintain a neutral pH and provide a dilute nutrient source for the suspension of microorganisms.
Automatically generated - may contain errors
2 protocols using buffered 0.1 peptone water
1
Microbiological Analysis of Food Samples
2
Microbiological Characterization of Hake Muscle
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Samples were analyzed, before and immediately after storage, as described by Salgado, López-Caballero, Gómez-Guillén, Mauri, and Montero (2013) . Briefly, 10 g of hake muscle was collected in a vertical laminar-flow cabinet (mod. AV 30/70 Telstar, Madrid, Spain) and introduced in a sterile plastic bag (Sterilin, Stone, Staffordshire, UK) with 90 mL of buffered 0.1% peptone water (Oxoid, Basingstoke, UK). After 1-minute processing in a Stomacher blender (model Colworth 400, Seward, London, UK), ten-fold serial dilutions were prepared in buffered peptone water and duplicates of the dilutions were plated on the appropriate medium, according to the procedures that follow.
Total aerobic mesophiles (TAM) were determined by the pour plate method on Plate Count Agar (PCA, Oxoid). Plates were incubated at 30 °C for 72 h and the colonies formed were counted. Enterobacteriaceae (ENT) were quantified on double-layered plates of Violet Red Bile Glucose agar (VRBG, Oxoid) after incubation at 30 °C for 48 h. Plate counts were expressed as the decimal logarithm of colony-forming units (CFU) per gram of hake loin (log 10 CFU•g -1
). The detection limit was 1 log 10 CFU•g -1 in both cases.
Total aerobic mesophiles (TAM) were determined by the pour plate method on Plate Count Agar (PCA, Oxoid). Plates were incubated at 30 °C for 72 h and the colonies formed were counted. Enterobacteriaceae (ENT) were quantified on double-layered plates of Violet Red Bile Glucose agar (VRBG, Oxoid) after incubation at 30 °C for 48 h. Plate counts were expressed as the decimal logarithm of colony-forming units (CFU) per gram of hake loin (log 10 CFU•g -1
). The detection limit was 1 log 10 CFU•g -1 in both cases.
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