Barbosa, J.

Fraqueza, M. J.

Patarata, L.

Rezende, L. Pinto de

Teixeira, P.

English

Repositório Institucional da Universidade Católica Portuguesa

Organized by:Food Development & ProductionConclusions• There is considerable variability between different lots of Salpicão.• Nitrites have an unquestionable impact on the microbial composition of the product. • Sensorial and stability alterations might occur due to the impact of nitrites on the microbiota.• Lactic Acid Bacteria dominate the microbiota of the later  processing stages.• Listeria decreases throughout processing, possibly due to LAB bacteriocins.• Products without nitrites have heightened levels of Clostridium.0%10%20%30%40%50%60%70%80%90%100%Classification at Genus levelDesulfomonileEnterobacterThermodesulfobiumLachnoclostridiumSodalisPsychromonasSerratiaStreptococcusWigglesworthiaGloeomargaritaMoritellaColwelliaAnoxybacillusKlebsiellaHaemophilusAlistipesLactococcusBacteroidesPhotobacteriumPsychrobacterPediococcusShewanellaVibrioAeromonasAcinetobacterCarnobacteriumLeuconostocPseudomonasScytonemaBrochothrixWeissellaLactobacillusLot 1Impact of nitrite on the microbiological quality of a Portuguese Salpicão• Pinto de Rezende L1, Barbosa J1, Patarata L2, Fraqueza MJ3, Teixeira P1*• 1 Universidade Católica Portuguesa, CBQF - Centro de Biotecnologia e Química Fina – Laboratório Associado, Escola Superior de Biotecnologia, Rua Diogo Botelho 1327, 4169-005 Porto, Portugal• 2 CIISA, Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, Av. da Universidade Técnica, Pólo Universitário, Alto da Ajuda,1300-477, Lisbon, Portugal• 3 CECAV, Animal and Veterinary Research Center, Universidade de Trás-os-Montes e Alto Douro, 5001-801 Vila Real, Portugal• *Email: pcteixeira@ucp.ptMethodsSamples of each phase of production (raw meat T0; before stuffing T1; middle of curing process T2; finished product T3; middle of shelf-life T4; and end of shelf-life T5) were collected and totalDNA was extracted following the Qiagen DNeasy® mericon® food kit protocol. This extraction was performed independently in two different lots. Amplification of the 16s rRNA gene, specifically thehypervariable V3/V4 region, was performed. PCR products were sequenced by the Illumina MiSeq® sequencer.Results and Discussion• Significant variation between lots was observed.• Bacteria of the Lactobacillus genus dominated themicrobiota of the later phases in both samples.• The reduction observed in Listeriaceae presencemight be a result of the production of bacteriocins byLactobacillus species.• Lactic Acid Bacteria cultures as starters could hinderpathogenic proliferation.• Pseudomonas, Lactobacillus, and Weissella were notaffected in the presence of nitrite.• Brochothrix, and Salmonella appeared susceptible tonitrite activity.• Clostridium was found in lower abundance insamples with nitrite (0.02%) than in nitrite-freesamples (0.23%).• Nitrite seems to be effective in preventingClostridium spp. growthReferences• 1. Karwowska M, Stadnik J, Stasiak DM, Wójciak K, Lorenzo JM. Strategies to improve the nutritional value of meat products: incorporation ofbioactive compounds, reduction or elimination of harmful components and alternative technologies. Int J Food Sci Technol. 2021;56(12):6142-6156.doi:10.1111/IJFS.15060• 2. Shakil MH, Trisha AT, Rahman M, et al. Nitrites in Cured Meats, Health Risk Issues, Alternatives to Nitrites: A Review. Foods 2022, Vol 11,Page 3355. 2022;11(21):3355. doi:10.3390/FOODS11213355• 3. Sebranek JG, Bacus JN. Cured meat products without direct addition of nitrate or nitrite: what are the issues? Meat Sci. 2007;77(1):136-147.doi:10.1016/J.MEATSCI.2007.03.025• 4 Majou D, Christieans S. Mechanisms of the bactericidal effects of nitrate and nitrite in cured meats. Meat Sci. 2018;145:273-284.doi:10.1016/J.MEATSCI.2018.06.013• 5. Dürre P. Physiology and Sporulation in Clostridium. Microbiol Spectr. 2014;2(4). doi:10.1128/MICROBIOLSPEC.TBS-0010-2012• 6. Kalaycloǧlu Z, Erim FB. Nitrate and Nitrites in Foods: Worldwide Regional Distribution in View of Their Risks and Benefits. J Agric FoodChem. 2019;67(26):7205-7222. doi:10.1021/ACS.JAFC.9B01194/SUPPL_FILE/JF9B01194_SI_001.PDFIntroductionThe development of healthier traditional product formulations is required by new trends in the food industry1,2. The curing of meats is disseminated throughout the world, being firmly established intraditional gastronomies. Meat curing is widespread throughout the world and is an integral part of traditional gastronomy. The use of nitrite salts is paramount in suppressing lipid oxidation and reducegrowth of pathogenic bacteria. This activity is especially significant when inhibition of Clostridium botulinum2 is necessary, inhibiting vegetative cell proliferation and germination of spores3,4,5. While this iswidely recognised, the impact of nitrite on the total microbiota of fermented meats is not yet fully understood. The ingestion of excessive nitrite has been correlated with an increased risk of cardiovascularpathologies, diabetes, blood diseases, and cancers2,6. As a result, a popular adverse reaction to the use of nitrite salts has been trending6.With NGS (Next-Generation Sequencing) techniques, detecting changes in microbial communities can be performed with unparallel specificity. Therefore, this work aimed to analyse the development ofmicrobiological characteristics, as a result of nitrite elimination, throughout the various processing phases of Salpicão.Acknowledgements• We would like to thank Centre for Interdisciplinary Research in Animal Health - CIISA (Project UIDP/ CVT/00276/2020, funded by FCT). Funding: This work was financially supported by Safe Meat Products Project “Quality & Safety of Processed Meat Products” (PDR2020-1.0.1- FEADER-031359). Financial support for the author L. Pinto de Rezende was provided by the doctoral fellowship 2022.13871.BDLot 20%10%20%30%40%50%60%70%80%90%100%Classification at Family levelRuminococcaceaeSyntrophaceaeThermodesulfobiaceaePectobacteriaceaePsychromonadaceaeGloeomargaritaceaeMoritellaceaeFlavobacteriaceaeYersiniaceaeColwelliaceaeLachnospiraceaeBacillaceaeErwiniaceaePasteurellaceaeRikenellaceaeStreptococcaceaeBacteroidaceaeEnterobacteriaceaeShewanellaceaeAeromonadaceaeVibrionaceaeMoraxellaceaeCarnobacteriaceaePseudomonadaceaeScytonemataceaeListeriaceaeLeuconostocaceaeLactobacillaceaeLot 1 Lot 2T0 T1 T2 T3 T4 T5 T0 T1 T2 T3 T4 T5- + - + - + - + - + - + - + - + - + - +LactobacillaceaeLeuconostocaceaeListeriaceaeScytonemataceaePseudomonadaceaeCarnobacteriaceaeMoraxellaceaeVibrionaceaeAeromonadaceaeShewanellaceaeEnterobacteriaceaeBacteroidaceaeStreptococcaceaeRikenellaceaePasteurellaceaeErwiniaceaeBacillaceaeLachnospiraceaeColwelliaceaeYersiniaceaeFlavobacteriaceaeMoritellaceaeGloeomargaritaceaePsychromonadaceaePectobacteriaceaeThermodesulfobiaceaeSyntrophaceaeRuminococcaceaeT0 T1 T2 T3 T4 T5 T0 T1 T2 T3 T4 T5- + - + - + - + - + - + - + - + - + - +LactobacillusWeissellaBrochothrixScytonemaPseudomonasLeuconostocCarnobacteriumAcinetobacterAeromonasVibrioShewanellaPediococcusPsychrobacterPhotobacteriumBacteroidesLactococcusAlistipesHaemophilusKlebsiellaAnoxybacillusColwelliaMoritellaGloeomargaritaWigglesworthiaStreptococcusSerratiaPsychromonasSodalisLachnoclostridiumThermodesulfobiumEnterobacterDesulfomonileFigure 1 – Heatmap of the total microbiota, at the Family level. Legend: Green – High density; Red – Low density; White – Percentile 50; (+) - with nitrite; (-) – without nitriteFigure 3 – Heatmap of the total microbiota, at the Genus level. Legend: Green – High density; Red – Low density; White – Percentile 50; (+) - with nitrite; (-) – without nitriteFigure 2 – Variation of the total microbiota throughout processing, with and without nitrite, at the Genus level. Figure 4 – Variation of the total microbiota throughout processing, with and without nitrite, at the Genus level. 

Impact of nitrite on the microbiological quality of a Portuguese Salpicão

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Impact of nitrite on the microbiological quality of a Portuguese Salpicão

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Repositório Institucional da Universidade Católica Portuguesa