) by optical density (periodic absorbance readings) at 620 nm in culture media Brain Heart Infusion broth (BHI, HiMedia, India). Throughout the growth curve, cell counts were determined selleck compound as log CFU/ml by serial dilution in peptone water 0.1% (w/v) and subsequent enumeration on Brain Heart Infusion agar (BHI agar, HiMedia, India) by

a spread plate methodology. C. perfringens spores were quantified during the growth curve by a Most Probable Number (MPN) method previously described by Scott et al. (2001). Dried aerial parts of winter savory spice (S. monatana L.) originating from Albania (Mediterranean climate country and mountainous region located in Southeastern Europe on the Balkan peninsula 41°21′N and 19°59′ W), were acquired from a spice store (Mr. Josef Herbs and Spices) at the local market city of São Paulo (SP, Brazil). The EO was extracted by hydrodistillation using a modified Clevenger apparatus. Dry plant material was placed with water in a 6000 ml volumetric distillation flask.

The flask was coupled to the modified Clevenger apparatus, and the extraction was performed for 3 h with the temperature maintained at 100 ± 5 °C. The obtained hydrolate (water/oil fraction) was centrifuged at 321.8 g for 10 min at 25 °C. The EO was collected with a Pasteur pipette, and the water traces selleck products were removed with anhydrous sodium sulfate (Vetec, Brazil). The oil was stored under refrigeration temperature (5 ± 2 °C) in glass flasks wrapped in aluminum foil ( Guimarães et al., 2008). Aerial parts of the winter savory (5 g) were placed with 80 ml cyclohexane (Vetec, Brazil) in a 250 ml volumetric distillation flask. The flask was coupled to a condenser with a graduated volumetric

collector and heated at 100 ± 5 °C for 2 h. After the distillation process, the volume of water in the collector was measured and expressed as the moisture content contained per 100 g sample. For the yield calculation, 350 g of dry spice was subjected to extraction by hydrodistillation, and the EO obtained was quantified. In parallel to the moisture content measurement, the EO yield for dried plants was obtained (% w/w) as the moisture free basis (MFB) (Pimentel et al., 2006). The EO chemical components were identified by gas chromatography coupled to mass spectrometry (GC–MS). Sclareol A Shimadzu gas chromatograph (model GC 17A) equipped with a mass selective detector (model QP 5000) was operated under the following conditions: fused silica capillary column (30 m × 0.25 mm) coated with a DB-5 MS stationary phase; ion source temperature of 220 °C; column temperature programmed at an initial temperature of 40 °C, and increased by 3 °C/min up to 240 °C; helium carrier gas (1 ml/min); initial column pressure of 100.2 kPa; split ratio of 1:10 and volume injected of 1 μl (1% solution in dichloromethane). The following conditions were used for the mass spectrometer (MS): impact energy of 70 eV; decomposition velocity of 1000, decomposition interval of 0.