, 1998, Burt, 2004 and Gutierrez et al., 2009). EOs are volatile, natural and complex compounds characterized by a strong odor and are formed by aromatic plants as secondary metabolites. Galunisertib concentration In addition to being used as flavoring agents in foods, EOs exhibit antibacterial, antifungal and antioxidant properties (Bakkali et al., 2008). Satureja montana L., commonly known as winter savory or mountain savory, belongs to the Lamiaceae family, Nepetoideae subfamily and Mentheae tribe and is a perennial semi-shrub (20 cm–30 cm) that inhabits arid, sunny and rocky regions. S. montana
L. is native to the Mediterranean and is found throughout Europe, Russia and Turkey. S. montana L. is a strong aromatic herb and has been used for centuries as a spice for food and teas; is used in Mediterranean cooking, mainly as a seasoning for meats and fish and in flavoring agents for soups, sausages, canned meats and spicy sauces ( Slavkovska et al., 2001, Mastelić and Jerković, PARP inhibitor 2003, Bezbradica et al., 2005, Ćetković et al., 2007 and Silva et al., 2009). S. montana L. has biological properties that are related to the presence of its major EO chemical compounds thymol and carvacrol ( Radonic and Milos, 2003 and Mirjana and Nada, 2004). This research was aimed to evaluate the antimicrobial effect of winter savory (S. montana L.) EO (0.0%, 0.78%, 1.56% and 3.125%) on C. perfringens type A (ATCC
3624) added in mortadella-type sausages formulated with different levels of NaNO2 (0 ppm, 100 ppm Endonuclease and 200 ppm) stored at 25 °C for 30 days. This study was also aimed to determine the feasibility of reducing the levels of nitrite used in the product formulation through the combined use of savory EO to control C. perfringens. The bacterium used in this research was C. perfringens type A ATCC
3624 (history I.C. Hall: L.S. Mc Clung 1997; A.J. Wildson type A, cep26; INCQS 00053), which was kindly provided by the National Institute of Quality Control in Health (INCQS) of the Oswaldo Cruz Foundation (Rio de Janeiro, Brazil). The bacterial strain was reactivated in a specific medium semisolid Clostridium Broth (Biolife Italiana Srl, Italy) under anaerobic conditions at 37 °C for 24 h. After the strain grew, the bacterial cells were pelleted by centrifugation (5000 g for 5 min at 24 °C), covered by freezing culture medium (15% glycerol Vetec, Brazil; 0.5% bacteriological peptone and 0.3% of yeast extract, Biolife Italiana Srl, Italy; and 0.5% of NaCl, final pH of 7.2 ± 0,2) and maintained under a freezing temperature (− 20 °C) throughout the experiment. For bacterial reactivation and use, an aliquot of the freezing culture medium was transferred to test tubes containing the Clostridium Broth medium and grown with two subcultures (last in Brain Heart Infusion broth) at 37 °C for 24 h under anaerobic conditions. The standardization of cell counts was carried out by the growth curve.