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Evaluation of a bacteriocinogenic Lactobacillus plantarum strain on the microbiological characteristics of “Alheira de Vitela”

1 Universidade Católica Portuguesa, CBQF—Centro de Biotecnologia e Química Fina—Laboratório Associado, Escola Superior de Biotecnologia, Rua Arquiteto Lobão Vital 172, 4200-374 Porto, Portugal
2 Bioinformatics Unit, Department of Infectious Diseases, National Institute of Health, 1649-016 Lisbon, Portugal

Topical Sections: Food microbiology and food safety

Lactic Acid Bacteria (LAB) and their bacteriocins can be successfully used as natural preservatives in meat products. This work aimed to investigate the effect of fresh and lyophilized starter cultures of an autochthonous bacteriocinogenic LAB strain (Lactobacillus plantarum ST153Ch: bac + culture) on the microbiological characteristics of “Alheira”, a traditional Portuguese smoked product. “Alheira” with the addition of fresh or lyophilized culture (ca. 108 cfu/g) and “Alheira” control (no bacteriocinogenic culture added) were produced by an industrial meat company. The antilisterial activity of this culture in this food matrix was investigated, with some samples being inoculated with Listeria monocytogenes (ca. 105 cfu/g). Detection of L. monocytogenes, Salmonella spp., sulphite reducing clostridia, Yersinia enterocolitica and enumeration of L. monocytogenes, Staphylococcus aureus, Bacillus cereus, Escherichia coli, Enterobacteriaceae, lactic acid bacteria, yeasts and moulds were performed immediately after production and at 3, 7, 15, 21, 28, 60 and 90 days of storage at 4 ℃, according to ISO methodologies. Also, a 16S rRNA Gene Analysis was performed of the microbial communities of “Alheira” with and without the lyophilized bacteriocinogenic culture. Pathogenic and indicator organisms were not detected or were below acceptable levels in all samples. LAB counts increased during storage and reached similar values after 15 days (ca. 1010 cfu/g) in all samples. There was a clear trend for a higher reduction of L. monocytogenes in the presence of the bioprotective culture, more pronounced during the initial 15 days of storage. From the analysis of the microbial communities of samples of “Alheiras” at different stages of fermentation, Leuconostocaceae and Lactobacillaceae predominated in all the samples and Lactobacillus was the genus more prevalent in “Alheiras” after 60 days of storage with the addition of bacteriocinogenic culture.
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1. Albano H, 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.    

2. Esteves A, Patarata L, Saraiva C, et al. (2008) Assessment of the microbiological characteristics of industrially produced alheira, with particular reference to foodborne pathogens. ‎J Food Saf 28: 88–102.    

3. 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.    

4. Felício M, Ramalheira R, Ferreira V, et al. (2011) Thermal inactivation of Listeria monocytogenes from alheiras, traditional Portuguese sausage during cooking. Food Control 22: 1960–1964.    

5. Campos CA, Castro MP, Rivas FP, et al. (2013) Bacteriocins in food: Evaluation of the factors affecting their effectiveness. In: Mendéz-Vilas A, Microbial pathogens and strategies for combating them: science, technology and Education, Ed., Formatex, Badajoz, 994–1004.

6. Oliveira M, Ferreira V, Magalhães R, et al. (2018) Biocontrol strategies for Mediterranean-style fermented sausages. Food Res Int 103: 438–449.    

7. Orihuel A, Bonacina J, Vildoza MJ, et al. (2018) Biocontrol of Listeria monocytogenes in a meat model using a combination of a bacteriocinogenic strain with curing additives. Food Res Int 107: 289–296.    

8. Perez R, Zendo T, Sonomoto K (2014)Novel bacteriocins from lactic acid bacteria (LAB): various structures and applications. Microb Cell Fact 13: S3.

9. 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.    

10. Mainar MS, Xhaferi R, Samapundo S, et al. (2016) Opportunities and limitations for the production of safe fermented meats without nitrate and nitrite using an antibacterial Staphylococcus sciuri starter culture. Food Control 69: 267–274.    

11. 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.

12. Todorov SD, Vaz-Velho M, Franco BDGM, et al. (2013) Partial characterization of bacteriocins produced by three strains of Lactobacillus sakei, isolated from salpicão, a fermented meat product from North-West of Portugal. Food Control 30: 111–121.    

13. Vaz-Velho M, Jácome S, Noronha L, et al. (2013) Comparison of antilisterial effects of two strains of lactic acid bacteria during processing and storage of a Portuguese salami-like product "Alheira". Chem Eng Transactions 32: 1807–1812.

14. Bergholz TM, Switt AIM, Wiedmann M (2014) Omics approaches in food safety: fulfilling the promise? Trends Microbiol 22: 275–281.    

15. Ferrocino I, Bellio A, Giordano M, et al. (2018) Shotgun metagenomics and volatilome profile of the microbiota of fermented sausages. Appl Environ Microbiol 84:1–14.

16. Kergourlay G, Taminiau B, Daube G, et al. (2015) Metagenomic insights into the dynamics of microbial communities in food. Int J Food Microbiol 213: 31–39.

17. Briers Y, Klumpp J, Schuppler M, et al. (2011) Genome Sequence of Listeria monocytogenes Scott A, a clinical isolate from a food-Borne listeriosis outbreak. J Bacteriol 193: 4284–4285.    

18. Barbosa J, Borges S, Amorim M, et al. (2015) Comparison of spray drying, freeze drying and convective hot air drying for the production of a probiotic orange powder. J Funct Foods 17: 340–351.    

19. ISO 15214:1998. Food Microbiology - Horizontal method for the enumeration of mesophilic lactic acid bacteria - Colony-count technique at 30 degrees C.

20. ISO 21528-2:2017. Microbiology of the food chain-Horizontal method for the detection and enumeration of Enterobacteriaceae - Part 2: Colony-count technique.

21. 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.

22. ISO 6888-2:1999. Microbiology of food and animal feeding stuffs - Horizontal method for the enumeration of coagulase-positive staphylococci (Staphylococcus aureus and other species)-Part 2: Technique using rabbit plasma fibrinogen agar medium.

23. 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.

24. ISO 7932:2004. Microbiology of food and animal feeding stuffs-Horizontal method for the enumeration of presumptive Bacillus cereus - Colony-count technique at 30 degrees C.

25. NP 3277-1:1987. Food Microbiology: Moulds and Yeast colony counting-Part 1: Incubation at 25 °C.

26. ISO 11290-1:2017. Microbiology of the food chain - Horizontal method for the detection and enumeration of Listeria monocytogenes and of Listeria spp.-Part 1: Detection method.

27. ISO 11290-2:2017. Microbiology of the food chain - Horizontal method for the detection and enumeration of Listeria monocytogenes and of Listeria spp.-Part 2: Enumeration method.

28. ISO 6579-1:2017. Microbiology of the food chain-Horizontal method for the detection, enumeration and serotyping of - Part 1: Detection of Salmonella spp.

29. NP 2262:1986. Food Microbiology. General guidelines for sulfite-reducing Clostridium spores detection.

30. ISO 10273:2017. Microbiology of the food chain-Horizontal method for the detection of pathogenic Yersinia enterocolitica.

31. Herlemann DP, Labrenz M, Jürgens K, et al. (2011) Transitions in bacterial communities along the 2000 km salinity gradient of the Baltic Sea. The ISME J 5: 1571–1579.    

32. Klindworth A, Pruesse E, Schweer T, et al. (2013) Evaluation of general 16S ribosomal RNA gene PCR primers for classical and next-generation sequencing-based diversity studies. Nucleic Acids Res 41: e1.    

33. Illumina MiSeq System (2013) 16S metagenomic sequencing library preparation. Preparing 16S ribosomal RNA gene amplicons for the Illumina MiSeq System.

34. Comeau AM, Douglas GM, Langille MGI (2017) Microbiome Helper: a custom and streamlined workflow for microbiome research. mSystems 2: e00127-16.

35. Wood DE, Salzberg SL (2014) Kraken: ultrafast metagenomic sequence classification using exact alignments. Genome Biol 15: R46.    

36. Foegeding PM, Thomas AB, Pilkington DH, et al. (1992) Enhanced control of Listeria monocytogenes by in situ-produced pediocin during dry fermented sausage production. Appl Environ Microbiol 58: 884–890.

37. Campanini M, Pedrazzoni I, Barbuti S, et al. (1993) Behaviour of Listeria monocytogenes during the maturation of naturally and artificially contaminated salami: effect of lactic acid bacteria starter cultures. Int J Food Microbiol 20: 169–175.    

38. Hugas M, Garriga M, Aymerich MT, et al. (1995) Inhibition of Listeria in dry fermented sausage by the bacteriocinogenic Lactobacillus sake CTC494. J Appl Microbiol 79: 322–330.

39. Mataragas M, Drosinos EH, Metaxopoulos J (2003) Antagonistic activity of lactic acid bacteria against Listeria monocytogenes in sliced cooked cured pork shoulder stored under vacuum or modified atmosphere at 4 ± 2 ºC. Food Microbiol 20: 259–265.    

40. Nieto-Lozano JC, Reguera-Useros JI, Peláez-Martínez MC, et al. (2006) Effect of a bacteriocin produced by Pediococcus acidilactici against Listeria monocytogenes and Clostridium perfringens on Spanish raw meat. Meat Sci 72: 57–61.    

41. Albano H, Pinho C, Leite D, et al. (2009) Evaluation of a bacteriocin-producing strain of Pediococcus acidilactici as a biopreservative for "Alheira", a fermented meat sausage. Food Control 20: 764–770.    

42. Ercolini D, De Filippis F, La Storia A, et al. (2012) ''Remake'' by high throughput sequencing of the microbiota involved in the production of water buffalo mozzarella cheese. App Environ Microbiol 78: 8142–8145.    

43. De Pasquale I, Calasso M, Mancini L, et al. (2014) Causal relationship between microbial ecology dynamics and proteolysis during manufacture and ripening of protected designation of origin (PDO) cheese Canestrato Pugliese. Appl Environ Microbiol 80: 4085–4094.    

44. Połka J, Rebecchi A, Pisacane V, et al. (2015) Bacterial diversity in typical Italian salami at different ripening stages as revealed by high-throughput sequencing of 16S rRNA amplicons. Food Microbiol 46: 342–356.    

45. Jackson CR, Randolph KC, Osborn SL, et al. (2013) Culture dependent and independent analysis of bacterial communities associated with commercial salad leaf vegetables. BMC Microbiol 13: 274.    

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