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Redalyc.Spoilage and Pathogenic Bacteria Isolated from Two Types Of Ciencia y Tecnología Alimentaria ISSN: 1135-8122 [email protected] Sociedad Mexicana de Nutrición y Tecnología de Alimentos México Matos, J. S. T.; Jensen, B. B.; Barreto, S. F. H. A.; Hojberg, O. Spoilage and pathogenic bacteria isolated from two types of portuguese dry smoked sausages after shelf-life period in modified atmosphere package Ciencia y Tecnología Alimentaria, vol. 5, núm. 3, 2006, pp. 165-174 Sociedad Mexicana de Nutrición y Tecnología de Alimentos Reynosa, México Available in: http://www.redalyc.org/articulo.oa?id=72450301 How to cite Complete issue Scientific Information System More information about this article Network of Scientific Journals from Latin America, the Caribbean, Spain and Portugal Journal's homepage in redalyc.org Non-profit academic project, developed under the open access initiative SOMENTA Cienc. Tecnol. Aliment. 5(3) 165-174 (2006) CIENCIA Y Sociedad Mexicana de Nutrición www.somenta.org/journal ISSN 1135-8122 TECNOLOGÍA y Tecnología de los Alimentos ALIMENTARIA SPOILAGE AND PATHOGENIC BACTERIA ISOLATED FROM TWO TYPES OF PORTUGUESE DRY SMOKED SAUSAGES AFTER SHELF-LIFE PERIOD IN MODIFIED ATMOSPHERE PACKAGE BACTERIAS DETERIORATIVAS Y PATÓGENAS AISLADAS DE DOS TIPOS DE CHORIZOS DESPUÉS DEL PERIODO DE VIDA DE ANAQUEL ENVASADAS EN ATMÓSFERAS MODIFICADAS Matos, J. S. T.1#*; Jensen, B. B.1; Barreto, S. F. H. A.2; Hojberg, O.1 1 Danish Institute of Agricultural Sciences, Department of Animal Health, Welfare and Nutrition, 8830 Tjele, Denmark. 2 Faculdade de Medicina Veterinária, CIISA, UTL, R. Prof. Cid dos Santos - Polo Universitário, Alto da Ajuda, 1300- 477 Lisboa, Portugal. #Present address: Instituto Superior de Agronomia; Departmento de Produção Agrícola e Animal - Secção de Produção Animal, Tapada da Ajuda, 1349-017 Lisboa, Portugal. Recibido/Received: 13-08-2006; aceptado/accepted: 07-11-2006 *Autor para la correspondencia. E-mail: [email protected] Abstract Members of the bacterial population of two types of Portuguese dry smoked sausages (chouriços) were identified and characterized. A total of 77 isolates were recovered from chouriços type Alentejano and Ribatejano, after 120 days at 20 ± 5 ºC in modified atmosphere package (55 % N2/45 % CO2) using non-selective and selective media. Based on phenotypic (cell morphology and fermentation profile) and genotypic (16SrDNA sequencing) characters, the isolates were grouped into the following genera: Bacillus, Enterococcus, Clostridium, Staphylococcus, Pediococcus, Lactobacillus and Paenibacillus. The species identified were: Enterococcus faecium (24.7 %), Bacillus subtilis (23.4 %), Staphylococcus epidermidis (14.3 %), B. cereus (7.8 %), Pediococcus acidilactici (6.5 %) Bacillus pumilus (6.5 %), Clostridium bifermentans (3.9 %), B. licheniformis, and Enterococcus faecalis (2.6 %). Major differences observed between the two varieties of chouriço were the absence of Pediococcus species and the presence of high numbers of isolates identified as Staphylococcus epidermidis in chouriço type Alentejano and, the high incidence of Bacillus species in sausage type Ribatejano. Differences found between products may be due to spices formulation in each type of product, the type of natural casings used and, the hygienic quality of the raw meat. Resumen Se identificaron y caracterizaron los miembros de la población bacteriana de dos tipos de chorizos portugueses. Un total de 77 aislados fueron recuperados de los chouriços tipo Alentejano y Ribatejano, después de 120 días de almacenamiento con empaque en atmósfera modificada (55 % N2/ 45 % CO2) a 20 ± 5 °C usando medios de cultivo selectivos y no selectivos. Con base en caracteres del fenotipo (morfología celular y perfil de fermentación) y del genotipo (secuencia del 16SrDNA) los aislados fueron agrupado en los siguientes géneros: Bacilos, Enterococos, Clostridium, Staphylococos, Pediococos, Lactobacilos y Paenibacilos. Las especies identificadas fueron: Enterococcus faecium (24,7 %), Bacillus subtilis (23,4 %), Staphylococcus epidermidis (14,3 %), B. cereus (7,8 %), Pediococcus acidilactici (6,5 %) Bacillus pumilus (6,5 %), Clostridium bifermentans (3,9 %), B. licheniformis, and Enterococcus faecalis (2,6 %). Las principales diferencias observadas entre las dos variedades de chouriço fueron la ausencia de especies de Pediococcus y la presencia de un elevado número de aislados identificados como Staphylococcus epidermidis en chouriço tipo Alentejano y la alta incidencia de especies de Bacillus en el chouriços tipo Ribatejano. Las diferencias encontradas entre los productos pueden deberse a las especias incluidas en la formulación en cada tipo de producto, el tipo de recubrimiento natural y de la calidad higiénica de la carne cruda. Keywords: Chouriço, modified atmosphere package, shelf life, bacteria, identification Palabras clave: Chorizo, envase en atmósferas modificadas, vida de anaquel, bacteria, identificación INTRODUCTION one very short maturation period, varying from one to three days. During this time all the meat, spices and curing Portuguese dry smoked sausage (chouriço) is very ingredients are held at temperatures below 10 ºC. After this different from Spanish and other dry fermented sausages, period the ingredients are stuffed into natural casings not only in its taste but also in the processing technology. (salted pork intestine or beef dry casing), intensively The Spanish sausage has a similar composition but is smoked at high temperature for about 3 days. It is then exposed to a long dry period at low temperature and is considered a smoked sausage processed by heat (Sousa almost not smoked. Portuguese chouriço is exposed to and Ribeiro, 1983). 165 Cienc. Tecnol. Aliment. 5(3) 165-174 (2006) ISSN 1135-8122 ©2006 SOMENTA The microbiology of dry smoked sausages is smoked dry sausages consisted of a mixture of pork meat variable and complex and the rate of spoilage of these meat and fat minced (approx. 20 mm) with the formulation products can reduce the shelf life and cause substantial ingredients red pepper paste, water, salt, garlic paste, olive financial losses to manufacturing companies. The spoilage oil, spices, white wine, sugar, liquid smoke and additives ecology is affected by a combination of intrinsic and (sodium nitrite and commercial sausage sodium extrinsic factors (Korkeala and Makela, 1989). Factors such polyphosphate) as outlined in Table 1. as ripening technique (Cantoni and Comi, 1986), use of Major differences between the two products natural intestines of pork and beef to stuff meat batters consisted in olive oil, spices, hot red pepper paste and (Byun et al., 2001), bacterial population of raw meat material white wine, besides the casings used. This paste is matured (Nel et al., 2004; Sardinha et al., 2000), introduction of for one day at 5 ºC. The stuffing is filled into salted pork spices as formulation ingredients (Banerjee and Sarkar, intestine in the Ribatejano sausage type and into beef dry 2003a), smoke (Wendorff, 1981), application of modified casing for the Alentejano sausage type and, subject to atmosphere and the CO2 concentrations (Stiles, 1991; the drying effect of smoke and temperature for about three Sivertsvik et al., 2002) and, storage temperature (Korkeala days. The experiments were conducted at a commercial and Makela, 1989) all have significant influence on the meat plant and the study was based on 12 samples (each microflora of these types of meat products and also on sample was composed by a mixture of three sausages, a the growth and survival of spoilage microorganisms during total of 36 sausages) randomly drawn from two batches. shelf-life period. Representative sausages of each experiment were packed Several studies (Encinas et al., 1996; Garcia-Varona separately in modified atmosphere (55 % N2/45 % CO2) with et al., 2000; te Giffel et al., 1996; Cocolin et al., 2001; a net weight of about 225g and stored at 20 ± 5 ºC for 120 Daporta Padin, 1988; Santos et al., 1998; Papamanoli et days (shelf-life period determined by the manufacturing al., 2003; Hugas et al., 2003; Devriese et al., 1995) were industry) for further analysis. Representative sausage conducted to identify bacterial populations on fermented samples presented pH values after production of 5.80 ± dry sausages, however, despite increasing interest for 0.17 and of 5.70 ± 0.10 and water activity varied between modified atmosphere package (MAP), the effect of MAP 0.93 ± 0.01 and 0.94 ± 0.01 [mean values ± standard on the spoilage bacteria of these meat products is not yet deviation (n=6)] for sausage type A and R, respectively. clear. Besides identifying the bacterial contamination After storage, pH values varied from 5.60 ± 0.00 and 5.43 ± during the manufacture processing, it is further important 0.06 for sausage type A and R, respectively, and water to identify the spoilage microorganisms that will remain or activity remained with the same values (data not shown). proliferate during the storage period in MAP, and might decrease shelf life compromising product safety. The type Isolation of bacteria of spoilage microorganisms are primarily related to the Microbial analyses were performed on triplicate extent and type of the initial contamination as affected by samples. Each sample was prepared from three sausages. the resident «factory flora», the manufacturing site and Twenty five grams were removed (approximately 8 g from the processing hygiene. The spoilage potential is not only each sausage) were cut in to pieces of 1 cm and dependent on the bacterial growth rates, but also on
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