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- Pathogens-in-Foods (PIF): An open-access European database of occurrence data of biological hazards in foodsPublication . Gonzales-Barron, Ursula; Faria, Ana Sofia; Thebault, Anne; Guillier, Laurent; Mendes, Lucas Ribeiro; Silva, Lucas Ribeiro; Messens, Winy; Kooh, Pauline; Cadavez, VascoThe collection of occurrence data of foodborne pathogens in foods faces the hindrances of dispersion of information, lack of standardisation and harmonisation, and ultimately, high expenditure in time and resources. The Pathogens-in-Foods (PIF) database was conceived as a solution to centralise published data on prevalence and concentration of pathogenic bacteria, viruses and parasites occurring in foods, obtained through systematic review (SR), and categorised in harmonised data structures under controlled terminologies. The present article outlines how PIF was constructed to adhere to the FAIR (findability, accessibility, interoperability and reusability) principles for scientific data management; and proceeds with a description of the PIF concept, which entails two phases: the SR process and the population of PIF. The protocolled SR process is supported by a welldefined search strategy, inclusion criteria, and rules for internal validation assessment; whereas the population of PIF with new data relies in data extraction, validation and release. The article then introduces a novel data quality approach, named as the CCC approach (data consistency, conformity and completeness), which ensures proper interpretation of data, richness of data, and flawless transcription of data. After a brief explanation of the three PIF components – database, back-end and front-end – the article proceeds with the exposition of the data model, as well as the capabilities of the front-end, including data search, insertion and curation. The future of PIF lies in expanding its capabilities, addressing emerging challenges, and leveraging technological advancements to maintain its relevance and utility in the evolving landscape of food safety.
- Microbiological and Physicochemical Profile of Traditionally Produced Chouriça de Carne Dry-Fermented Sausages: Towards Benchmarking of Products Against Established Quality GroupsPublication . Faria, Ana Sofia; Bonilla-Luque, Olga María; Carvalho, Laís; Fernandes, Nathália; Prieto Lage, Miguel A.; Cadavez, Vasco; Gonzales-Barron, UrsulaThe physicochemical and microbiological properties of traditional Portuguese ready-to-eat dry fermented sausage chouriça de carne samples from 14 regional producers were analysed and subjected to multivariate analysis to determine the relationships between them and to evaluate how the quality and safety of these sausages is affected by these properties. Producer-mean values for physicochemical analyses were quite variable, with intervals of 4.87–6.11 for pH, 0.803–0.965 for aw, moisture 19.5–48.5%, protein 32.0–60.1% (db), fat 22.0–53.3% (db), ash 3.52–9.69% (db), and carbohydrates 1.66–13.5% (db). Mesophilic counts varied (5.61–8.68 log CFU/g), while lactic acid bacteria were generally high (MRS: 8.21–10.2; M17: 7.66–10.0 log CFU/g). S. aureus was enumerated in levels up to 2.55 log CFU/g, while presumptive C. perfringens never surpassed 2 log CFU/g. Salmonella spp. and Listeria spp. were also detected in the samples tested. Principal component (PC) analysis yielded a three-dimension solution that explained 60% of the data variation; PC1 (26%) characterized chorizo formulations with more meat, while PC2 (19.3%) described sausages with longer/rapid fermentation, and PC3 (14.5%) highlighted chouriços with poorer hygiene. Cluster analysis identified three quality groups: (i) chorizos with high moisture, high protein content, and lowest pH; (ii) sausages with low moisture, high fat content, and elevated pH; and (iii) chouriças with high moisture and high protein but lower fat contents, low pH, and improved hygiene. Lastly, factor analysis yielded a varimax-rotated three-factor solution that explained 65% of the data, with similar results to PCA; factor 1 (23.5%) depicted chorizos with low pH but high moisture, factor 2 (20.8%) described sausages with more meat in the formulation, and factor 3 (20.6%) longer or rapid fermentation. Overall, the results evidenced the great variability in the quality attributes of Portuguese chorizo sausages, very likely to arise from multiple recipes, ingredients, and manufacturing practices. The definition of quality clusters is expected to play a crucial role for the self-denominated “artisanal” food companies to benchmark their chouriço sausages against the proper artisanal quality group.
- Leptospiral antibodies in sera of fish from the Ave river: preliminary resultsPublication . Nunes-Pereira, Margarida; Varandas, Simone; Teixeira, Amílcar; Faria, Ana Sofia; Cabral, João Alexandre; Cortes, Rui M.V.; Paiva-Cardoso, Maria das NevesLeptospirosis is a zoonosis with world distribution, caused by infection with pathogenic spirochetes of the genus Leptospira. Leptospires’ natural habitat is the kidney of infected animals, being disseminated via urine to the environment, where they survive outside the host in fresh water or moist soils. Thus, leptospirosis is associated with activities such as fish farming, animal and agricultural production and water-related activities. The aim of this work was to detect leptospiral antibodies (Abs) in wild fish species (Luciobarbus bocagei, Pseudochondrostoma duriense and Squalius carolitertii) captured in the north of Portugal. Sera of 29 fish caught in the Ave River basin in July 2017 were tested by Microscopic Agglutination Test (MAT). Abs were detected in 51.7% (15/29) specimens, with reactivities against serogroups Icterohaemorrhagiae, Pomona, Canicola, Sejroe, Australis and Ballum. The highest Abs titre was 1:160, observed to Icterohaemorrhagiae serovar. Leptospiral Abs were detected in 75% (9/12) of P. duriense and in 46.2% (6/13) of L. bocagei. The results undeniably show that the sampled fish inhabit fresh water contaminated with leptospires and develop immune response, being potential sources of infection for man and animals. Further studies are needed to better understand potential public health threats.
- Effect of Sage (Salvia officinalis L.) Extract on the Survival of Staphylococcus aureus in Portuguese Alheira Sausage during MaturationPublication . Coelho-Fernandes, Sara; Rodrigues, Gisela; Faria, Ana Sofia; Caleja, Cristina; Pereira, Eliana; Pinela, José; Carocho, Márcio; Barros, Lillian; Cadavez, Vasco; Gonzales-Barron, UrsulaThe objective of this study was to assess the effect of sage (Salvia officinalis L.) extract on the survival of Staphylococcus aureus in a Portuguese non-ready-to-eat meat product (alheira sausage). Alheira batter was produced, mixed with 0.0%, 0.5%, 1.0%, or 1.5% of lyophilised sage extract, and stuffed in natural casings. Sausages were then individually inoculated with S. aureus and left to ferment/mature at 10 °C/85% RH for 10 days. Sage extract was found to inactivate S. aureus (p < 0.001) with no significant differences between doses. At the 10th day of maturation, S. aureus decreased in 1.146 log CFU/g (SE = 0.065 log CFU/g) in alheiras mixed with 0.5–1.0% sage extract. Nonetheless, this extract retarded the growth of indigenous lactic acid bacteria during maturation. The higher the dose, the greater the effect (p < 0.001).
- Staphylococcus aureus inactivation in a non-ready-to-eat sausage during maturation: a dynamic modelPublication . Coelho-Fernandes, Sara; Rodrigues, Gisela; Faria, Ana Sofia; Cadavez, Vasco; Gonzales-Barron, UrsulaThe aim of this study was to model the effect of annatto (Bixa orellana) extract against S. aureus (SA) in a Portuguese non-ready-to-eat meat product (alheira sausage) during maturation. Alheira batter was prepared with 0.0%, 0.5%, 1.0%, or 1.5% of lyophilised annatto extract and stuffed in pre-washed natural casings following inoculation with SA, then hung in a climatic-controlled chamber at 10 °C for 13 days. For every treatment, a dynamic model was adjusted and adequately fitted to all survival curves with residuals and root mean square errors between 0.0008–0.0016 and 0.029–0.040, respectively, producing significant parameter estimates. Therefore, the addition of annatto extracts significantly shortened the shoulder phase and decreased the time to achieve one log reduction, which, in practical terms, corresponded to up to 1.35 [SE = 0.08] log CFU/g reduction by the end of the 13-day maturation.
- Microbiological and physicochemical assessment of artisanally produced “Alheira” fermented sausages in Northern PortugalPublication . Coelho-Fernandes, Sara; Zefanias, Odete; Rodrigues, Gisela; Faria, Ana Sofia; Fernandes, Ângela; Barros, Lillian; Cadavez, Vasco; Gonzales-Barron, UrsulaAlheira is a traditional non-ready-to-eat sausage produced mainly in northern Portugal. The objective of this study was to evaluate the associations between some relevant physicochemical and microbiological attributes of alheiras produced by different regional producers. Finished products from 8 regional factories amounting to 40 samples were analyzed. Counts of mesophiles, lactic acid bacteria, Staphylococcus aureus, presumptive Clostridium perfringens, and Salmonella spp., as well as pH, water activity (aW), and proximate analysis were determined. Principal component analysis (PCA) of these variables was conducted to construct quality maps. Three meaningful components were extracted, accounting for 63% of data variability. PC1 (26% data variability) was positively associated with LAB, mesophiles, and S. aureus, characterizing therefore longer fer- mentation. PC2 (22% data variability) correlated negatively with moisture, aW and positively with C. perfringens, and thus has been linked to greater dehydration of sausages. PC3 (15% data varia- bility) correlated positively with pH and protein content, implying the use of more meat in the formulation. This preliminary work has identified three quality factors underpinning the variabil- ity in artisanal alheiras; and has also highlighted the need to implement better microbiological control and process standardization during the production of artisanal alheiras.
- Technological potential of lactic acid bacteria isolated from Portuguese goat’s raw milk cheesesPublication . Silva, Beatriz Nunes; Faria, Ana Sofia; Cadavez, Vasco; Teixeira, José António; Gonzales-Barron, UrsulaIn this work, a total of 97 MRS-grown and 135 M17-grown lactic acid bacteria strains were isolated from 20 goat’s raw milk cheeses, and their antimicrobial, acidifying, and proteolytic activities were determined in vitro. Principal component analyses adjusted to a subset of 84 promising isolates evidenced that, for MRS isolates, antagonism against Staphylococcus aureus correlated well with higher acidification potential, whereas for M17 isolates, the antagonisms against S. aureus and Listeria monocytogenes were more correlated. The outcomes highlighted various strains with pathogen inhibition ability and satisfactory technological properties that may be useful for the development of a customised starter culture.
- Screening of Lactic Acid Bacteria Isolated from Artisanally Produced Alheira Fermented Sausages as Potential Starter CulturesPublication . Faria, Ana Sofia; Fernandes, Nathália; Cadavez, Vasco; Gonzales-Barron, UrsulaAlheira is a traditional Portuguese fermented sausage, produced without the aid of any starter culture. The objective of this study was to isolate lactic acid bacteria (LAB) naturally present in alheiras, and screen their potential for use as functional starter. Forty alheiras from 8 regional producers were analysed. A total of 335 LAB were isolated from MRS and M17 media and confirmed. Antimicrobial activity against Staphylococcus aureus and Salmonella spp. at 37°C and 10°C was evaluated by spot-on-lawn assay. Sixty-three strains were pre-selected, and their lactic acid production, acidification capacity and proteolytic activity determined. The suitability of the strains was assessed by adjusting two separate Principal Component Analyses (PCA) on MRS and M17 data. Results showed acidification potential as the most determinant feature for strain differentiation across both media, followed by antimicrobial activity; while lactic acid production (LAC) and proteolytic activity (PAct) were the least contributing variables. For MRS strains, PC1 (38.9%) was positively correlated with the acidification capability of strains. PC2 (20.1%) positively correlated with LAC and negatively correlated with inhibition of both pathogens at 10°C and PAct, implying higher antimicrobial potential. Likewise, for M17 strains, PC1 (43.1%) and PC2 (16.7%) characterised higher acidification and greater antimicrobial potential, respectively. The results differentiated 3 clusters of MRS strains, one with higher acidification capacity, related to greater S. aureus inhibition at 37°C; a second cluster with better overall antimicrobial activity and linked to higher proteolytic activity; and a third cluster of strains with more rapid production of lactic acid. One standalone MRS strain presented both greater acidification and antimicrobial potential. From M17 isolates, only two strains presented the highest acidification ability and pathogenic inhibition. These results highlighted several strains with great potential for use as starter cultures in “Alheira”, which could confer protection against foodborne pathogens. In-situ essays are ongoing.
- Molecular identification of lactic acid producing bacteria isolated from alheira, a traditional Portuguese fermented sausagePublication . Fernandes, Nathália; Faria, Ana Sofia; Carvalho, Laís; Choupina, Altino; Rodrigues, Carina; Cadavez, Vasco; Gonzales-Barron, UrsulaPortuguese traditional fermented meat products constitute a valued economic and cultural heritage. The objective of this work was to screen the lactic acid bacteria (LAB) present in thealheira. 25 LABs were identified by Sanger sequencing of the 16S ribosomal gene. Sequencing results were aligned with sequences from the NCBI database using the BLAST algorithm. Genetic analysis showed a diverse lactic acid producing microbiome, and LABs from the family Lactobacillaceae and Leuconostocaceae were dominant, found in 64% of samples, while other organisms of the family Streptococcaceae and Enterococcaceae were found in 36% of samples. This work enabled the identification of LAB normally present in a traditional Portuguese product, as well as the desired technological characteristics that they can bestow to the product.
- Pathogens-in-Foods database: A web application for assessing the occurrence data of microbiological hazards in foods marketed in EuropePublication . Faria, Ana Sofia; Winter, Maiara; Thebault, Anne; Guillier, Laurent; Sanaa, Moez; Kooh, Pauline; Cadavez, Vasco; Gonzales-Barron, UrsulaPathogens-in-Foods (PIF) is a dynamic database constructed using systematic literature searches of occurrence data (prevalence and enumeration) of important pathogenic agents (Bacillus cereus, Campylobacter spp., Clostridium perfringens, Listeria monocytogenes, Salmonella spp., Shiga toxin-producing Escherichia coli, Staphylococcus aureus, Yersinia enterocolitica, Cryptosporidium spp., Giardia spp., Toxoplasma gondii, Hepatitis A virus, Hepatitis E virus and Norovirus) in foods randomly surveyed across Europe. After filtering the primary studies, these were screened for relevance and methodological quality, and the data were extracted into the PIF database following a systematic categorisation of microbiological methods, food types and outcomes. The database is freely accessible through a web application that facilitates data retrieval according to several relevant variables. The PIF spans data published from 2000 onwards and is intended for use by researchers and food authorities after meta-analysis, in microbiological risk assessment.