Research article Topical Sections

Microbiological characterization of different formulations of alheiras (fermented sausages)

  • Received: 31 December 2018 Accepted: 10 May 2019 Published: 24 May 2019
  • Different ingredients in old recipes are becoming popular and the traditional alheira did not escape to this new trend. The objective of this preliminary study was to characterize microbiologically nine different formulations of alheira from five producers. In this sense, isolates obtained were characterized through different phenotypic and biochemical tests. Their susceptibility to different antimicrobials and the presence of virulence factors was also investigated. Lactic acid bacteria were the predominant microbiota, but pathogenic bacteria as coagulase-positive staphylococci, Listeria monocytogenes and Salmonella spp. as well as indicator organisms were also found. Several virulence factors were produced among the different groups of isolates, with a high incidence of isolates producing β-haemolysis. Along with their potential pathogenic activity, also several antimicrobial resistances were found being the majority of isolates classified as multi-resistant. At our knowledge, this is the first study with these new formulations of alheira. A higher number of products must be analyzed, but we believe that results obtained in this study should help to alert consumers for the need of safe cooking time/temperatures of these products.

    Citation: Julieta Silva, Joana Barbosa, Helena Albano, Maria Sequeira, Ana Pinto, Conceição Costa Bonito, Margarida Saraiva, Paula Teixeira. Microbiological characterization of different formulations of alheiras (fermented sausages)[J]. AIMS Agriculture and Food, 2019, 4(2): 399-413. doi: 10.3934/agrfood.2019.2.399

    Related Papers:

  • Different ingredients in old recipes are becoming popular and the traditional alheira did not escape to this new trend. The objective of this preliminary study was to characterize microbiologically nine different formulations of alheira from five producers. In this sense, isolates obtained were characterized through different phenotypic and biochemical tests. Their susceptibility to different antimicrobials and the presence of virulence factors was also investigated. Lactic acid bacteria were the predominant microbiota, but pathogenic bacteria as coagulase-positive staphylococci, Listeria monocytogenes and Salmonella spp. as well as indicator organisms were also found. Several virulence factors were produced among the different groups of isolates, with a high incidence of isolates producing β-haemolysis. Along with their potential pathogenic activity, also several antimicrobial resistances were found being the majority of isolates classified as multi-resistant. At our knowledge, this is the first study with these new formulations of alheira. A higher number of products must be analyzed, but we believe that results obtained in this study should help to alert consumers for the need of safe cooking time/temperatures of these products.


    加载中


    [1] Ojha KS, Kerry JP, Duffy G, et al. (2015) Technological advances for enhancing quality and safety of fermented meat products. Trends Food Sci Technol 44: 105–116. doi: 10.1016/j.tifs.2015.03.010
    [2] Marcos C, Viegas C, De Almeida AM, et al. (2016) Portuguese traditional sausages: different types, nutritional composition, and novel trends. J Ethnic Foods 3: 51–60. doi: 10.1016/j.jef.2016.01.004
    [3] Albano H, Oliveira M, Aroso R, et al. (2007) Antilisterial activity of lactic acid bacteria isolated from 'Alheiras' (traditional Portuguese fermented sausages): in situ assays. Meat Sci 76: 796–800. doi: 10.1016/j.meatsci.2007.01.019
    [4] ISO 4833-1: 2013. Microbiology of the food chain - Horizontal method for the enumeration of microorganisms-Part 1: Colony count at 30 ℃ by the pour plate technique.
    [5] ISO 15214: 1998. Microbiologie des aliments-Méthode horizontale pour le enombrement des bactéries lactiques mésophiles-Technique par comptage des colonies à 30 ℃.
    [6] Ferreira V, Barbosa J, Vendeiro S, et al. (2006) Chemical and microbiological characterization of alheira: A typical Portuguese fermented sausage with particular reference to factors relating to food safety. Meat Sci 73: 570–575. doi: 10.1016/j.meatsci.2006.02.011
    [7] ISO 21528-2: 2000. Microbiology of food and animal feeding stuffs - Horizontal method for the detection and enumeration of Enterobacteriaceae-Part 2: colony-count method.
    [8] ISO 6888-1: 1999. Microbiology of food and animal feeding stuffs-Horizontal method for the enumeration of coagulase-positive staphylococci (Staphylococcus aureus and other species)-Part 1: Technique using Baird-Parker agar medium.
    [9] ISO 16649-2: 2001. Microbiology of food and animal feeding stuffs-Horizontal method for the enumeration of beta-glucuronidase-positive Escherichia coli-Part 2: Colony-count technique at 44 degrees C using 5-bromo-4-chloro-3-indolyl beta-D-glucuronide.
    [10] NP 3277-1: 1987. Food Microbiology: Moulds and Yeast colony counting-Part 1: Incubation at 25 ℃.
    [11] ISO 11290-1: 1996. Microbiology of food and animal feeding stuffs - Horizontal method for the detection and enumeration of Listeria monocytogenes - Part 1: Detection method.
    [12] ISO 6579: 2002. Microbiology of food and animal feeding stuffs-Horizontal method for the detection of Salmonella spp..
    [13]   NP 2262: 1986. IPQ food microbiology. General rules for identifying reducing Clostridium sulphite spores.
    [14] Albano H, van Reenen CA, Todorov SD, et al. (2009) Phenotypic and genetic heterogeneity of lactic acid bacteria isolated from 'Alheira', a traditional fermented sausage produced in Portugal. Meat Sci 82: 389–398. doi: 10.1016/j.meatsci.2009.02.009
    [15] Holt JG, Krieg NR, Sneath PHA, et al. (1994) Bergey's Manual of determinative bacteriology, 9th edition, Lippincott Williams & Wilkins.
    [16] Iorio NLP, Ferreira RBR, Schuenck RP, et al. (2007) Simplified and reliable scheme for species-level identification of Staphylococcus clinical isolates. J Clin Microbiol 45: 2564–2569. doi: 10.1128/JCM.00679-07
    [17] CDC (1998) Botulism in the United States. Handbook for epidemiologists, clinicians, and laboratory workers. Centers for disease control and prevention. National center for infectious diseases. Division of bacterial and mycotic diseases.
    [18] Clinical and Laboratory Standards Institute (2012) Performance Standards for Antimicrobial Susceptibility Tests; Document M100. Wayne, PA: Clinical and Laboratory Standards Institute.
    [19] European Food Safety Authority (2012) Guidance on the assessment of bacterial susceptibility to antimicrobials of human and veterinary importance by EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP). EFSA J 10: 2740–2749.
    [20] Barbosa J, Magalhães R, Santos S, et al. (2013) Evaluation of antibiotic resistance patterns of food and clinical Listeria monocytogenes isolates in Portugal. Foodborne Pathog Dis 10: 861–866. doi: 10.1089/fpd.2013.1532
    [21] Tiago I, Teixeira I, Silva S, et al. (2004) Metabolic and genetic diversity of mesophilic and thermophilic bacteria isolated from composted municipal sludge on poly-e-caprolactones. Curr Microbiol 49: 407–414. doi: 10.1007/s00284-004-4353-0
    [22] Ben Omar N, Castro A, Lucas R, et al. (2004) Functional and safety aspects of enterococci isolated from different Spanish foods. Syst Appl Microbiol 27: 118–130. doi: 10.1078/0723-2020-00248
    [23] Barbosa J, Gibbs PA, Teixeira P (2010) Virulence factors among enterococci isolated from traditional fermented meat products produced in the North of Portugal. Food Control 21: 651–656. doi: 10.1016/j.foodcont.2009.10.002
    [24] Bover-Cid S, Holzapfel WH (1999) Improved screening procedure for biogenic amine production by lactic acid bacteria. Int J Food Microbiol 53: 33–41. doi: 10.1016/S0168-1605(99)00152-X
    [25] Barbosa J, Borges S, Teixeira P (2014) Selection of potential probiotic Enterococcus faecium isolated from Portuguese fermented food. Int J Food Microbiol 191: 144–148. doi: 10.1016/j.ijfoodmicro.2014.09.009
    [26] Ferreira V, Barbosa J, Silva J, et al. (2007) Characterisation of alheiras, traditional sausages produced in the North of Portugal, with respect to their microbiological safety. Food Control 18: 436–440. doi: 10.1016/j.foodcont.2005.11.011
    [27] Govaris A, Botsoglou E, Sergelidis D, et al. (2011). Antibacterial activity of oregano and thyme essential oils against Listeria monocytogenes and Escherichia coli O157:H7 in feta cheese packaged under modified atmosphere. LWT Food Sci Technol 44: 1240–1244. doi: 10.1016/j.lwt.2010.09.022
    [28] Zakpaa HD, Imbeah CM, Mak-Mensah EE (2009) Microbial characterization of fermented meat products on some selected markets in the Kumasi metropolis, Ghana. Afr J Food Sci 3: 340–346.
    [29] Giraffa G. (2002) Enterococci from foods. FEMS Microbiol Rev 26: 163–171. doi: 10.1111/j.1574-6976.2002.tb00608.x
    [30] Barbosa J, Ferreira V, Teixeira P (2009) Antibiotic susceptibility of enterococci isolated from traditional fermented meat products. Food Microbiol 26: 527–532. doi: 10.1016/j.fm.2009.03.005
    [31] Ribeiro T, Oliveira M, Fraqueza MJ, et al. (2011) Antibiotic resistance and virulence factors among enterococci isolated from chouriço, a traditional Portuguese dry fermented sausage. J Food Prot 74: 465–469. doi: 10.4315/0362-028X.JFP-10-309
    [32] Flores M, Corral S, Cano-García L, et al. (2015) Yeast strains as potential aroma enhancers in dry fermented sausages. Int J Food Microbiol 12: 16–24.
    [33] Felício MTS, Hogg T, Gibbs P, et al. (2007) Recurrent and sporadic Listeria monocytogenes contamination in alheiras represents considerable diversity, including virulence-attenuated isolates. App Environ Microbiol 73: 3887–3895. doi: 10.1128/AEM.02912-06
    [34] Felício MTS, Ramalheira R, Ferreira V, et al. (2011) Thermal inactivation of Listeria monocytogenes from alheiras, traditional Portuguese sausage during cooking. Food Control 22: 1960–1964. doi: 10.1016/j.foodcont.2011.05.011
    [35] Castro A, Santos C, Meireles H, et al. (2016) Food handlers as potential sources of dissemination of virulent strains of Staphylococcus aureus in the community. J Infect Public Health 9: 153–160. doi: 10.1016/j.jiph.2015.08.001
    [36] Friedman M, Henika PR, Levin CE, et al. (2007) Recipes for antimicrobial wine marinades against Bacillus cereus, Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella enterica. J Food Sci 72: M207–M213. doi: 10.1111/j.1750-3841.2007.00418.x
    [37] Jahan M, Krause DO, Holley RA (2013) Antimicrobial resistance of Enterococcus species from meat and fermented meat products isolated by a PCR-based rapid screening method. Int J Food Microbiol 163: 89–95. doi: 10.1016/j.ijfoodmicro.2013.02.017
    [38] Valenzuela AS, Ben-Omar N, Abriouel H, et al. (2009) Virulence factors, antibiotic resistance, and bacteriocins in enterococci from artisan foods of animal origin. Food Control 20: 381–385. doi: 10.1016/j.foodcont.2008.06.004
    [39] Fraqueza MJ (2015) Antibiotic resistance of lactic acid bacteria isolated from dry-fermented sausages. Int J Food Microbiol 212: 76–88. doi: 10.1016/j.ijfoodmicro.2015.04.035
    [40] Federici S, Ciarrocchi F, Campana R, et al. (2014) Identification and functional traits of lactic acid bacteria isolated from Ciauscolo salami produced in Central Italy. Meat Sci 98: 575–584. doi: 10.1016/j.meatsci.2014.05.019
    [41] Gevers D, Huys G, Swings J (2003) In vitro conjugal transfer of tetracycline resistance from Lactobacillus isolates to other Gram positive bacteria. FEMS Microbiol Lett 225: 125–130.
    [42] Marty E, Bodenmann C, Buchs J, et al. (2012) Prevalence of antibiotic resistance in coagulase-negative staphylococci from spontaneously fermented meat products and safety assessment for new starters. Int J Food Microbiol 159: 74–83. doi: 10.1016/j.ijfoodmicro.2012.07.025
    [43] Rebecchi A, Pisacane V, Callegari ML, et al. (2015) Ecology of antibiotic resistant coagulase-negative staphylococci isolated from the production chain of a typical Italian salami. Food Control 53: 14–22. doi: 10.1016/j.foodcont.2015.01.001
    [44] Pereira V, Lopes C, Castro A, et al. (2009) Characterization for enterotoxin production, virulence factors, and antibiotic susceptibility of Staphylococcus aureus isolates from various foods in Portugal. Food Microbiol 26: 278–282. doi: 10.1016/j.fm.2008.12.008
    [45] Miranda JM, Guarddona M, Vázquez BI, et al. (2008) Antimicrobial resistance in Enterobacteriaceae strains isolated from organic chicken, conventional chicken and conventional turkey meat: A comparative survey. Food Control 19: 412–416. doi: 10.1016/j.foodcont.2007.05.002
    [46] Todorov SD, Stojanovski S, Iliev I, et al. (2017) Technology and safety assessment for lactic acid bacteria isolated from traditional Bulgarian fermented meat product "lukanka." Braz J Microbiol 48: 576–586. doi: 10.1016/j.bjm.2017.02.005
    [47] Jimoh SO, Shittu AA, Morhason-Bello I (2013) Occurrence of virulence factor and extended spectrum beta lactamase in Enterobacteriaceae associated with ready-to-eat-fruits. Int J Biol Biol Sci 2: 083–087.
    [48] European Food Safety Authority (2011) Scientific opinion on risk based control of biogenic amine formation in fermented foods. Panel on Biological Hazards (BIOHAZ). EFSA J 9: 2393.
    [49] Latorre-Moratalla ML, Bover-Cid S, Talon R., et al. (2010) Distribution of aminogenic activity among potential autochthonous starter cultures. J Food Prot 73: 524–525. doi: 10.4315/0362-028X-73.3.524
    [50] Durlu-Özkaya F, Ayhan K, Vural N (2001) Biogenic amines produced by Enterobacteriaceae isolated from meat products. Meat Sci 58: 163–166. doi: 10.1016/S0309-1740(00)00144-3
    [51] Martín B, Garriga M, Hugas M, et al. (2006) Molecular, technological and safety characterization of Gram-positive catalase-positive cocci from slightly fermented sausages. Int J Food Microbiol 107: 148–158. doi: 10.1016/j.ijfoodmicro.2005.08.024
    [52] Latorre-Moratalla ML, Bover-Cid S, Veciana-Nogués MT, et al. (2012) Control of biogenic amines in fermented sausages: role of starter cultures. Front Microbiol 3: article 169.
  • Reader Comments
  • © 2019 the Author(s), licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0)
通讯作者: 陈斌, bchen63@163.com
  • 1. 

    沈阳化工大学材料科学与工程学院 沈阳 110142

  1. 本站搜索
  2. 百度学术搜索
  3. 万方数据库搜索
  4. CNKI搜索

Metrics

Article views(4445) PDF downloads(1439) Cited by(3)

Article outline

Figures and Tables

Figures(1)  /  Tables(3)

/

DownLoad:  Full-Size Img  PowerPoint
Return
Return

Catalog