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Listeria monocytogenes is a pathogen of considerable public health importance with a high case fatality. L. monocytogenes can grow at refrigeration temperatures and is of particular concern for ready-to-eat foods that require refrigeration. There is substantial interest in conducting and modeling shelf-life studies on L. monocytogenes, especially relating to storage temperature. Growth model parameters are generally estimated from constant-temperature growth experiments. Traditionally, first-order and second-order modeling (or primary and secondary) of growth data has been done sequentially. However, omnibus modeling, using a mixed-effects nonlinear regression approach, can model a full dataset covering all experimental conditions in one step. This study compared omnibus modeling to conventional sequential first-order/second-order modeling of growth data for five strains of L. monocytogenes. The omnibus model coupled a Huang primary model for growth with secondary models for growth rate and lag phase duration. First-order modeling indicated there were small significant differences in growth rate depending on the strain at all temperatures. Omnibus modeling indicated smaller differences. Overall, there was broad agreement between the estimates of growth rate obtained by the first-order and omnibus modeling. Through an appropriate choice of fixed and random effects incorporated in the omnibus model, potential errors in a dataset from one environmental condition can be identified and explored. This research was part funded by the Irish Department of Agriculture, Food, and Marine Research Funding Program, Project Reference 17/F/244, “Understanding Listeria monocytogrenes growth in food”. U. Gonzales-Barron and V. Cadavez are grateful to the EU PRIMA program and the Portuguese Foundation for Science and Technology (FCT) for funding the ArtiSaneFood project (PRIMA/0001/2018) and to FCT for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020). U. Gonzales-Barron acknowledges the national funding by FCT, P.I., through the Institutional Scientific Employment Program contract. info:eu-repo/semantics/publishedVersion

Hepatitis E virus (HEV) is one of the main causes of viral hepatitis infection worldwide. Sources of contamination can vary greatly according to geographical regions and HEV genotypes. HEV is endemic and responsible for large waterborne epidemics involving human HEV-1 or HEV-2 genotypes in regions with limited sanitation, in contrast to industrialized countries, where HEV is mainly a foodborne zoonosis involving HEV-3 and HEV-4 zoonotic genotypes. Limited data on HEV source attribution are available, and all possible sources and transmission pathways of HEV are not fully identified. A systematic review and a meta-analysis of case-control and cohort studies (including transversal studies) were performed to determine the main risk factors associated with sporadic hepatitis E infection. Suitable scientific articles were identified through a systematic literature search and subjected to a methodological quality assessment. From each study, odds-ratio (OR) measures were extracted/calculated, as well as study characteristics such as population type, design, and risk factor hierarchy. Mixed-effects meta-analyses models were adjusted by population type to appropriate data partitions. Seventy-seven cohort and case-control studies conducted between 1986 and 2016 and investigating risk factors in mixed population, susceptible population, and pregnant women, were included in this meta-analysis. Hepatitis E cases were defined with serological exams and differentiated whenever the serological exam is associated or not with symptoms. This meta-analysis identified the parenteral pathway (blood transfusion, tattooing or IV injection, dialysis or hemodialysis), and routes of infection related to contaminated water, animal contact (occupational exposure) and consumption of foods as relevant risk factors for hepatitis E infection. With regards to the role of food, as suspected and sometimes proven in several studies, pig meat, pork sausages, and game meat are identified as significant risk factors for HEV, in particular undercooked pig meat, or meat preparations containing pig liver. In addition, consumption of shellfish (oysters and mussels), in which HEV can accumulate when water is environmentally contaminated (from animal or human origin), is also associated with the detection of anti-HEV antibodies. The results of this meta-analysis show that symptomatic and infected cases share the most explainable risk factors, and are in agreement with recent studies conducted in Europe. This meta-analysis reveals that some sources such as consumption of insufficiently treated water, shellfish, or vegetables are under-investigated. Future case-control studies should include population at risk but under-investigated, such as transplant recipients, pregnant women and children, and investigate other potential sources of HEV. The authors would like to thank ANSES staff and the members of the ANSES Working Group on Source Attribution of Foodborne Diseases: Moez Sanaa, Laurence Watier, Jean Christophe Augustin, Frédéric Carlin, Julie David, Philippe Fravalo, Laurent Guillier, Nathalie Jourdan-Da Silva, Alexandre Leclercq, Lapo Mughini-Gras, Isabelle Villena. The authors are also grateful to the reviewers for their help in improving the manuscript quality. U. Gonzales-Barron and V. Cadavez are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020). U. Gonzales-Barron also thanks FCT, P.I., for the institutional scientific employment program. info:eu-repo/semantics/publishedVersion

Campylobacter spp. is an important causative agent of diarrheal illness worldwide. The disease is frequently associated with foodborne transmission, but other routes of exposure are increasingly recognized. A systematic review and meta-analysis of case-control studies were performed to determine the main risk factors associated with sporadic campylobacteriosis. Suitable scientific articles published up to March 2017 were identified through a systematic literature search and subject to methodological quality assessment. From each study, odds ratios (OR) as measures of association were extracted or calculated, as well as study characteristics such as study population, design, type of model used and risk factor categorization. Mixed-effects meta-analytical models were adjusted by population type to appropriate data partitions. From 4453 identified references, the quality assessment stage was passed by 71 case-control studies focusing on sporadic campylobacteriosis. The eligible studies were conducted between 1981 and 2012 and provided 1336 ORs for meta-analysis. The meta-analysis identified international travel (pooled OR=4.626), recent use of gastric antacids (pooled OR=2.911), occupational exposure to animals/carcasses (pooled OR=3.022), and food consumption (in particular raw or undercooked eggs, poultry, beef, and dairy) as the main risks factors for sporadic campylobacteriosis in the mixed population. In the child population, the main risk factors concerned environmental/animal transmission routes (e.g. drinking untreated water (pooled OR=3.261), exposure to recreational water (pooled OR=3.156), exposure to farm/rural environment (pooled OR=3.128), contact with farm animals (pooled OR=2.747), person-to-person transmission (pooled OR=2.736) and consumption of raw milk (pooled OR =2.603). The results of this meta-analysis highlight the importance of overlooked routes and vehicles of transmission (environment, animal contact, and other food vehicles) of Campylobacter that should be explored in dedicated studies. The authors would like to thank Anses staff and the members of the Anses Working Group on Source Attribution of Foodborne Diseases: Moez Sanaa, Laurence Watier, Jean Christophe Augustin, Frédéric Carlin, Laurent Guillier, Nathalie Jourdan-Da Silva, Alexandre Leclercq, Anne Thébault, Nicole Pavio, Isabelle Villena. U. Gonzales-Barron and V. Cadavez are grateful to the Foundation for Food Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO ( UID/AGR/00690/2019 ). U. Gonzales-Barron thanks the national funding by FCT , P.I., through the Institutional Scientific Employment Program contract. info:eu-repo/semantics/publishedVersion

International audience; Giardia duodenalis is an important source of gastroenteritis worldwide. Endemic cases have been described in developing and industrialized countries. We analyzed risk factors for sporadic giardiasis by a systematic review and a meta-analysis of the literature. From 72 studies, contact with an infected person, lack of personal hygiene, and attending a child daycare center were identified as risk factors in children and adults.Feco-oral transmission was significantly associated with exposure to human sewage/waste water, untreated drinking water and recreational waters. Travel abroad was a risk factor in industrialized countries. No handwashing before eating or preparing food, eating unwashed vegetables, or composite food were significant risk factors. Breastfeeding was a protective factor in developing countries. Interestingly, contact with pets was found as a significant risk factor in children in this meta-analysis. This could be explored in future studies with the comparison of the Giardia assemblage isolated from humans and pets. In the future, it would be interesting to investigate more precisely the type of water, vegetable, and whenever possible the method of preparation/treatment. Environmental and epidemiological investigations of specific risk factors by assemblages and types of Giardia spp. should be further studied. Finally, host factors in relationship with the severity and sequelae of giardiasis deserve future research.

International audience

Listeria monocytogenes is a pathogen of considerable public health importance with a high case fatality. L. monocytogenes can grow at refrigeration temperatures and is of particular concern for ready-to-eat foods that require refrigeration. There is substantial interest in conducting and modeling shelf-life studies on L. monocytogenes, especially relating to storage temperature. Growth model parameters are generally estimated from constant-temperature growth experiments. Traditionally, first-order and second-order modeling (or primary and secondary) of growth data has been done sequentially. However, omnibus modeling, using a mixed-effects nonlinear regression approach, can model a full dataset covering all experimental conditions in one step. This study compared omnibus modeling to conventional sequential first-order/second-order modeling of growth data for five strains of L. monocytogenes. The omnibus model coupled a Huang primary model for growth with secondary models for growth rate and lag phase duration. First-order modeling indicated there were small significant differences in growth rate depending on the strain at all temperatures. Omnibus modeling indicated smaller differences. Overall, there was broad agreement between the estimates of growth rate obtained by the first-order and omnibus modeling. Through an appropriate choice of fixed and random effects incorporated in the omnibus model, potential errors in a dataset from one environmental condition can be identified and explored.

In order to evaluate the welfare of lambs reared up to 4 months of age in different management systems in Europe, nine lamb meat farms (3 in Spain, 2 in Portugal, 1 in Germany, 1 in Slovenia and 2 in Italy) belonging to different production systems (intensive, semi-intensive, semi-extensive, extensive, shepherded) throughout Europe were visited. The first level welfare assessment according to the AWIN protocol for sheep was used, the data collection flow being as follows: (1) qualitative behaviour assessment (QBA), social withdrawal, stereotypy, excessive itching, panting; (2) fleece cleanliness, fleece quality, tail length, faecal soiling, lameness; (3) familiar human approach; (4) stocking density, access to shade/shelter, water availability; (4) lamb mortality. A great heterogeneity was observed because the breeds and the productive systems in which the animals have been raised are very different; only one of the purely extensive farms had no sheds for the animals, whereas the stocking density in confined animals was always appropriate. The results showed a high degree of compliance with the needs of the animals, which also corresponded to good productive results, but the objectification of the emotional state through the QBA yielded highly variable results, those animals reared under intensive and semiintensive farms grouped around positive descriptors, which may indicate that lambs reared in intensive conditions show a different and apparently calmer mental state than those reared in very extensive conditions. To the project EcoLamb (Holistic production to reduce the ecological footprint of meat), funded by ERA-Net SusAn (project ID 48) through the Spanish Agencia Estatal de Investigaci´on (PCIN-2017-074); to the Slovenian Ministry of Agriculture, Forestry and Food for financial support (Decision n◦ 631–10/2015/7), info:eu-repo/semantics/publishedVersion

In Portugal, the main autochthonous sheep breeds exploited for meat production are Churra Galega Bragancana (CGB) and Bordaleira de Entre Douro e Minho (BEDM). This study aimed to characterise the evolution of spoilage microorganisms in refrigerated vacuum-packed (VP) lamb meat from BEDM and CGB breeds and to elucidate how the intrinsic properties of meat can affect its microbial spoilage. Meat from the BEDM breed presented higher (p 0.05), the growth of spoilage bacteria was exacerbated by higher moisture (p < 0.0001) and higher ash content (p < 0.001). On the contrary, a higher intramuscular fat content slowed down (p < 0.0001) the growth of spoilage bacteria in VP lamb meat.

Alheira is a traditional non-ready-to-eat sausage produced mainly in Northern Portugal. Made of poultry, pork or game meat, bread and seasonings, it is a very sought-after meat product in Portuguese cuisine due to its organoleptic properties. 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 analysed. Counts of mesophiles, lactic acid bacteria (LAB on MRS and M17 agar), Staphylococcus aureus, Clostridium spp. and Salmonella spp., as well as pH, water activity (aW), moisture, ashes and protein content 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. PCA1 (26% of data) was positively associated with LAB, mesophiles and S. aureus, characterising therefore longer fermentation. PCA2 (22%) correlated negatively with moisture, aW and positively with Clostridium, and thus has been linked to greater dehydration of these sausages. PCA3 (15%) correlated positively with pH and protein content, implying the use of more meat in the formulation. Projection of samples in the three-dimensional space allowed the differentiation of three clusters of factories: one made up of 4 factories that represent microbiologically safe alheiras of lower pH and higher LAB. Another quality cluster of 3 factories describes meat-rich alheiras of higher pH and ash content. A third cluster of one factory was identified representing drier alheiras with higher incidence of pathogenic bacteria. This preliminary work has identified three quality factors (axes) underpinning the variability in artisanal alheiras: duration of fermentation, extent of dehydration and proportion of meat in formulation; and has also highlighted the need to implement better microbiological control and process standardisation during the production of artisanal alheiras.

Since Listeria monocytogenes has a high case-fatality rate, substantial research has been devoted to estimate its growth rate under different conditions of temperature, pH and water activity (aw). In this study, published findings on L. monocytogenes growth in broth were extracted and unified by constructing meta-regression models based on cardinal models for (i) temperature (CM[T]), (ii) temperature and pH (CM[T][pH]), and (iii) temperature, pH and aw (CM[T][pH][aw]). After assessing all the sources retrieved between 1988 until 2017, forty-nine primary studies were considered appropriate for inclusion. Apart from the modelling variables, study characteristics such as: type of broth (BHI, TSB, TPB), reading method (colony-forming-units, CFU; or binary-dilution optical density methods, OD), inoculum concentration and strain serotype, were also extracted. Meta-regressions based on CM[T] and CM[T][pH] were fitted on subsets of the 2009 growth rate measures and revealed that type of broth and reading method significantly modulated the cardinal parameter estimates. In the most parsimonious CM[T][pH][aw] meta-regression model, whereby the variability due to type of broth was extracted in a nested random-effects structure, the optimum growth rate ?opt of L. monocytogenes was found to be lower when measured as CFU (0.947 h-1; SE=0.094 h-1) than when measured as OD (1.289 h-1; SE=0.092 h-1). Such a model produced the following cardinal estimates: Tmin=-1.273°C (SE=0.179 °C), Topt=37.26°C (SE=0.688 °C), Tmax=45.12°C (SE=0.013 °C), pHmin=4.303 (SE=0.014), pHopt=7.085 (SE=0.080), pHmax=9.483 (SE=0.080), aw min=0.894 (SE=0.002) and aw opt=0.995 (SE=0.001). Integrating the outcomes from numerous L. monocytogenes growth experiments, this meta-analysis has estimated pooled cardinal parameters that can be used as reference values in quantitative risk assessment studies., Foundation for Food Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020). U. Gonzales-Barron acknowledges the national funding by FCT, P.I., through the Institutional Scientific Employment Program contract, info:eu-repo/semantics/publishedVersion

Listeriosis is a major public health concern associated with high hospitalization and mortality rates. The objective of this work was to summarize evidence on the associations between risk factors and sporadic cases by meta-analysing outcomes from currently published case-control studies. Suitable scientific articles were identified through systematic literature search, and subjected to a methodological quality assessment. From each study, odds-ratio (OR) measures as well as study characteristics such as population type, design, type of model and risk factor hierarchy were extracted. Mixed-effects meta-analysis models were adjusted by population type to appropriate data partitions. Twelve primary studies investigating sporadic listeriosis conducted between 1985 and 2013 passed through a quality assessment stage. These studies provided 226 OR considered for meta-analysis. According to the meta-analysis, the main risk factor for acquiring listeriosis is suffering from an immunocompromising disease. In relation to the food exposures, this meta-analysis confirmed known risk factors such as consumption of RTE dairy, seafood and processed meat and underlined new food vehicles as fruits and vegetables, recently involved in outbreaks. There were not enough data to appraise travel, animal-contact and person-to-person as transmission pathways for listeriosis. These results will allow refining the case-control studies in the aim of improving risk factors characterisation for listeriosis in the susceptible population. U. Gonzales-Barron and V. Cadavez are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020). U. Gonzales- Barron also thanks FCT, P.I., for the institutional scientific employment program. info:eu-repo/semantics/publishedVersion

In this study, the kinetic parameters of mesophilic, psychrotrophic and lactic acid bacteria in vacuum-packed beef at 1 °C and 4 °C were estimated from experimental growth curves produced by samples stored during 21 and 60 days, respectively. In a separate experiment, the survival of multidrug resistant (MDR) Salmonella enterica O:4,5 at 1°C was also characterized. The shelf-life of vacuum-packed beef stored at 4 °C was estimated at 16.1 days (95% CI: 14.8 – 17.3 days), whereas at 1 °C it was longer than 21 days because the mesophiles count estimated towards the end of the experiment was 12.5 ln CFU.g-1 (95% CI: 11.8 – 13.3 ln CFU.g-1) which is lower than the shelf-life reference value. At 1 °C, inoculated Salmonella was reduced in 6.61 ln CFU.g-1 (2.87 log CFU.g-1). These results demonstrated the importance of establishing in legislation, especially in Brazil, standard values of deteriorating microorganisms in beef for maintaining product quality. Neste estudo, os parâmetros cinéticos de bactérias mesófilas, psicrotróficas e ácido lácticas foram estimados em carne bovina embalada a vácuo a 1 °C e 4 °C, a partir de curvas experimentais produzidas em amostras estocadas durante 21 e 60 dias, respectivamente. Em um experimento separado, a sobrevivência de Salmonella enterica O:4,5 multirresistente (MDR) a 1°C também foi caracterizada. A vida de prateleira da carne bovina embalada a vácuo, estocada a 4°C, foi estimada em 16.1 dias (95% CI: 14.8 – 17.3 dias), enquanto que a 1 °C o período foi maior que 21 dias, porque a contagem estimada de mesófilos ao final do experimento foi de 12.5 ln UFC.g-1 (95% CI: 11.8 – 13.3 ln UFC.g-1), o qual é mais baixo que o valor referência de shelf-life. A 1 °C, Salmonella inoculada reduziu em 6.61 ln UFC.g-1 (2.87 log UFC.g-1). Estes resultados demonstram a importância de estabelecimento em legislação, especialmente no Brasil, de valores padrões para contagem de microrganismos deteriorantes em carnes visando manter a qualidade do produto. The authors thank the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Brazil, for supporting the first author with a scholarship from the International Sandwich Exchange Program (PDSE) approved at the Call 047/2017/Process: 88881.189927/2018- 01. We also thank the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Brazil (Process: 310462 / 2018-5), and the Instituto Federal de Educação, Ciência e Tecnologia de Mato Grosso (IFMT) for their support. Our gratitude also to the “Ad hoc” evaluators who reviewed our work and contributed to its improvement. info:eu-repo/semantics/publishedVersion

The ingestion of bioactive compounds has revealed health benefits, namely in the prevention and/or treatment of several diseases. This work aims to characterize the phenolic profile of three colour varieties of Chenopodium quinoa Willd. grains (black, red and white), and also evaluate their cytotoxic and antimicrobial activity. All varieties revealed the presence of phenolic compounds, namely, quercetin and kaempferol derivatives. In this study, quinoa grains did not reveal any anti-proliferative capacity in tumour cell lines, and, as expected, they were devoid of any toxicity. All of the analysed extracts possessed antibacterial and antifungal activities (inhibitory and bactericidal/fungicidal) against the microbial strains considered, exhibiting promising values of minimum bactericidal concentrations (mean MBC 0.153–0.916 mg/mL) and minimum fungicidal concentrations (mean MFC 0.211–0.884 mg/mL). Quinoa varieties represent a good source of bioactive compounds, interfering beneficially in the organism, specifically as antimicrobial agents. Thus, these extracts could be used in the development of bioactive ingredients. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020); FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E and TRANSCoLAB 0612_TRANS_CO_LAB_2_P; and L. Barros, R. Calhelha, and Gonzales-Barron thank the national funding by FCT, P.I., through the institutional scientific employment program contract. This work has also been supported by the Ministry of Education, Science and Technological Development of Republic of Serbia (451-03-68/2020-14/200007). info:eu-repo/semantics/publishedVersion

Although the nutritional profile, bioactivities, and uses of mesquite pod flour from various Prosopis species have been studied, limited research has been conducted on Prosopis pallida (Humb, & Bonpl. Ex Willd.) Kunth mesquite flour. This study aimed to characterize the nutritional quality and bioactive properties of P. pallida pod flour and to assess its technological performance in breadmaking as a partial replacer of white wheat flour. Peruvian P. pallida mesquite flour was found to have an appealing nutritional profile, with high contents of dietary fiber (29.6% dw) and protein (9.5% dw), and low contents of fat (1.0% dw) and carbohydrates (57.6% dw). It is a source of palmitic (12.6%), oleic (35.5%), and linoleic acids (45.8%), -, -, and - tocopherols, and contains phenolic compounds such as apigenin glycoside derivatives with proven antioxidant capacities. Extracts of P. pallida flour were also found to have antimicrobial and antifungal e ects and did not show hepatoxicity. When formulated as a wheat flour replacer, increasing mesquite flour levels yield composite doughs of lower stickiness and extensibility, and composite breads of lower elasticity (p < 0.01). However, up to a level of 10%, mesquite flour significantly increases loaf volume, reduces crumb hardness, and produces a more uniform crumb of small size alveoli (p < 0.01). Considering the purpose of improving the nutritional and technological quality of wheat flour bread, the addition of P. pallida pod flour can be highly recommended. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020). U.G.-B. L.B. and R.C.C. also thank the national funding by FCT—Foundation for Science and Technology, P.I., through the institutional scientific employment program contract. C.P. is grateful to the celebration of the program-contract foreseen in No. 4, 5 and 6 of article 23 of Decree-Law No. 57/29 August 2016, amended by Law No. 57/19 July 2017. This work has been supported by Ministry of Education, Science and Technological Development of Republic of Serbia (451-03-68/2020-14/200007). info:eu-repo/semantics/publishedVersion

Bio-preservation methods based on the use of natural extracts have been proposed as hurdles to increase the safety of many food products. The objective of this work was to study the phytochemical profile, biological activity, and antioxidant potential of six plant extracts, and their ability to be used as food preservatives. In this sense, rosemary, lemon balm, basil, tarragon, sage, and spearmint dry aerial parts were mechanically grinded. The extractions were performed using a sample/solvent ratio of 1:20, in a shaking water bath (at 150 rpm) at 60 °C for 90 min, and a Soxhlet apparatus (7 recycles), with ethanol 70% (v/v) (Et70%) and distilled water (dH2O) as solvents in both methods. The influence of the distinct extraction methods and solvents on the total phenolic content (TPC, determined by Folin-Ciocalteu method), total flavonoid content (TFC), and antioxidant activity (determined by FRAP, ABTS•+ and DPPH assays) of the natural extracts was evaluated. Identification and quantification of phenolic compounds was achieved by Ultra-High Performance Liquid Chromatography. The results show that both methodologies and solvents are adequate to obtain bioactive extracts. The TPC of water bath extracts was similar regardless of the solvent, apart from spearmint and sage extracts (higher TPC in ethanolic extracts). Alternatively, higher differences were observed in the TPC of Soxhlet extracts depending on the solvent, as aqueous extracts revealed greater TPC values. The TFC revealed differences due to the solvent for both methods: for water bath extracts, Et70% lead to higher TFC; while for Soxhlet extracts, dH2O delivered extracts with greater TFC. Regarding antioxidant activity, all extracts showed promising results. These outcomes provide insight on the phytochemical profile and biological activity of plant extracts and their value as bio-preservatives to be included in foods, thus encouraging the food industry to replace chemical preservatives with others that are plant-based.

The chemical composition and volatile profile of the Longissimus thoracis et lumborum muscle of lambs reared in two distinct production systems (intensive and extensive) was evaluated. For this, sixty-six lambs for meat production of the autochthonous Gallega Iberian breed were raised with concentrate and grass in intensive (30 animals) and extensive (36 animals) system, respectively, until 4–4.5 months of age when they were slaughtered. Subsequently, Longissimus thoracis et lumborum muscles were excised for the determination of chemical composition (moisture, intramuscular fat, protein, and ash percentages) and volatile substances. The aftermaths obtained evidenced that moisture (75.90% and 75.68%), intramuscular fat (1.68% and 1.76%), and protein (20.62% and 20.97%) contents were not significantly (P gt 0.05) affected by the production system. However, the extensively-fed lambs displayed a higher ash content (1.35% vs. 1.24%). Additionally, the total volatile content was also not significantly (P gt 0.05) influenced by feeding system. Despite this, the total content of hydrocarbons, acids, aldehydes, ketones, esters, ethers, furans, sulfur compounds, and others was significantly (P lt 0.05) affected by diet, being the alcohol family the only group not influenced (P gt 0.05) by the production system (1321.3 vs. 1211.3 AU × 104/g fresh muscle). Specifically, intensively-fed lambs showed significantly higher amounts for all volatile families apart from ketones (2215 vs. 2826 AU × 104/g fresh muscle) and sulfur compounds (22.7 vs. 123.7 AU × 104/g fresh muscle). In addition, benzyl alcohol and carbon disulfide were proposed as appropriate biomarkers for grass diets, while 1-butanone, 2-heptanone, and furan, 2-penthyl were indicated as suitable tracers for concentrate-based diets. The authors are grateful to EU ERA-NET program and the Portuguese Foundation for Science and Technology (FCT) and the Agencia Estatal de Investigación (Acciones de Programaci´on Conjunta Internacional, grant number PCIN-2017-053) for funding the project “EcoLamb–Holistic Production to Reduce the Ecological Footprint of Meat (SusAn/ 0002/2016). Authors would like to acknowledge to Consellería de Cultura, Educación e Ordenación Universitaria (Xunta de Galicia) for granting Noemí Echegaray with a predoctoral scholarship (Grant number IN606A-2018/002). Rubén Domínguez, Roberto Bermúdez Mirian Pateiro and José M. Lorenzo are members of the HealthyMeat network, funded by CYTED (ref. 119RT0568). CIMO authors are grateful to FCT and FEDER under Program PT2020 for financial support to CIMO (UIDB/00690/2020). Dr. Gonzales-Barron acknowledges the national funding by FCT, P.I., through the Institutional Scientific Employment Program contract. info:eu-repo/semantics/publishedVersion

This review compiles published information concerning the incidence of pathogenic microorganisms — Listeria monocytogenes, Salmonella spp., Staphylococcus aureus and shiga-toxin producing Escherichia coli (STEC) — in goat and sheep raw milk and cheese. Meta-analytical data were extracted from 37 primary studies undertaken in Australia, Brazil, China, Colombia, Costa Rica, Czech Republic, Egypt, Germany, Greece, Iran, Italy, Malaysia, Mexico, Norway, Poland, Portugal, Spain, Sweden, Switzerland, Turkey, UK and USA. Pooled frequencies of detection of pathogens were found to be similar for sheep and goat raw milk: Salmonella (1.4–2.4%), L. monocytogenes (2.9–3.6%), STEC (4.3–4.8%) and S. aureus (35–39%). Likewise, in goat cheeses, regardless of being made of raw or heat-treated milk, S. aureus has been the most frequent contaminant (16.0%), whereas in raw milk cheeses, regardless of origin, the pooled prevalence of S. aureus is equally high in hard (34.6%) and soft cheeses (25.7%). L. monocytogenes is another important pathogen in sheep and goat milk cheeses (3.6–12.8%) while E. coli O157 strains with virulence genes (4.3%) also appear to persist during cheese manufacture. As expected, STEC has a higher pooled incidence in raw milk cheeses (10.0%) than in pasteurised milk cheeses (4.7%). Thus, the moderate contamination in raw milk and cheese of sheep and goat origin, revealed by this meta-analysis, advocates the reinforcement of general prevention measures such as close monitoring of hygiene on farms and eradication of disease by sheep and goat dairy farmers. Moreover, for the production of traditional cheeses made of raw milk, preventive measures during processing, namely, regular sterilisation of dairy equipment, process monitoring and hygiene of operators, should be even more stringent.

The management of Heat Resistant Moulds (HRMs) is considered a great challenge for the juice fruit industry. Neosartorya, Byssochlamys and Talaromyces are three out of the main genera isolated from fruit juices that show great resistance to heat treatments. Several inactivation parameters can be found in the literature, however all of them were carried out in specific food matrices and using diverse inactivation methods. Thus, this meta-analysis study synthesizes the thermal resistance parameters of the three HRMs by adjusting extended Bigelow-based meta-regression models to data on inactivation experiments conducted in different liquid media. The meta-analytical data, extracted from publications between 1969 and 2017, was composed of decimal reduction time (D), inactivation method, temperature of inactivation, pH, °Brix, age of spores, and type of medium (model, juice, concentrates). Pooled D* values (D at 90 °C, pH 3.5 and 12° Brix) were estimated for B. fulva (1.95 min; 95% CI: 1.21–3.11 min), Talaromyces (4.03 min; 95% CI: 3.43–4.74 min), Neosartorya (0.5.35 min; 95% CI: 4.10–7.08 min), and B. nivea (10.32 min; 95% CI: 5.81–18.4 min). It was found that increasing the soluble solids in concentrates tends to cause a lower decrease in the heat resistance of Neosartorya and Talaromyces than increasing the soluble solids in model liquid or juices (p = 0.001; 0.012). In general, the screw-capped tubes and three neck round inactivation methods render higher D* values (p lt 0.05) than the thermal death tubes, the polyethylene bag and the capillary methods. Spores of Talaromyces (overall zpH = 7.56; 95% CI: 5.13–13.5) and Neosartorya (overall zpH = 7.07; 95% CI: 5.04–10.8) appear to be more thermal sensitive to a decrease in medium pH than spores of Byssochlamys (overall zpH = 4.34; 95% CI: 3.20–6.73). The meta-regression models presented in this study can be valuable for estimating pooled inactivation kinetic parameters to be used by the fruit juice industry in the management of thermal processes and in the determination of shelf-life. V.O. Alvarenga, L.P-Silva and A.S. Sant’Ana acknowledge CYTED Program (Network 115 RT0488) for the financial support. A.S. Sant’Ana acknowledges the financial support from CNPq (Grant #302763/2014- 7, #305804/2017-0) and to the Coordenaç˜ao de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001. U. Gonzales-Barron and V. Cadavez are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020). U. Gonzales- Barron also thanks the national funding by FCT, P.I., through the institutional scientific employment programme contract her contract. info:eu-repo/semantics/publishedVersion

The sustainability of the European sheep farming sector can be ensured through the high and consistent quality of products, although the concept of meat quality, being multifaceted, is challenging to keep up by farmers and producers. The objectives of this study were (i) to evaluate quality attributes (CIE L*, a*, b*, lipid oxidation, cooking loss and Warner-Bratzler texture test) of lamb meat during cold storage originating from eight European breeds raised under their normal production systems (n = 292 lambs); and (ii) to elucidate the influence of storage and intrinsic properties (cold carcass weight [CCW], ultimate pH [pH24], water activity [aw], and proximate composition) on the aforementioned quality attributes measured on the 3rd, 9th and 15th day post slaughter. All meat quality properties displayed a high variability due to the breed × production systems, although, as a whole, L*, a*, b*, TBARs and cooking loss increased during storage (P lt 0.001), whereas the tenderisation process was of non-linear decay nature (P = 0.001). While production system – taken across breeds – was not found to affect meat cooking losses, it heavily influenced CCW, producing intensive systems heavier (P lt 0.001) carcasses (21.4 kg) than those raised in semi-extensive (14.9 kg) and extensive systems (13.8 kg). On the 15th day of storage, TBARs from intensive systems was significantly (P lt 0.001) higher than those of extensively raised lambs (0.839 mg MDA/kg vs. 0.299 mg MDA/kg, respectively). Although the extensive and semi-extensive systems initially produced less tender meat (P lt 0.05) than intensive systems, on the 15th day of storage, meat from extensive (23.4 N/cm2) and semi-extensive systems (24.4 N/cm2) presented as good tenderness quality (P lt 0.10) as that of the intensive systems (22.1 N/cm2). Within breed × production systems, higher CCW or intramuscular fat was associated to darker meat (P lt 0.001 and P = 0.014), higher redness (P lt 0.001 for both), greater lipid oxidation (P = 0.016 and P lt 0.001), and lower slope (P = 0.014 and P = 0.004) and force (P = 0.027 and P = 0.001). The only intrinsic property that heavily affected most of the quality attributes was ash content, since meat of higher ash content presented higher luminosity (P = 0.008), and lower redness (P lt 0.001), yellowness (P lt 0.001), TBARS (P lt 0.001), cooking loss (P = 0.002) and tenderness (P lt 0.001). The elucidation of the impact of intrinsic properties on the lamb meat quality attributes during storage can help breeders and producers modify current flock management, feeding strategies and pre-slaughter and slaughter practices towards quality improvement or meeting particular consumers’ demands. The authors are grateful to EU ERA-NET programme and the Portuguese Foundation for Science and Technology (FCT) for funding the project “EcoLamb–Holistic Production to Reduce the Ecological Footprint of Meat (SusAn/0002/2016). CIMO authors are grateful to FCT and FEDER under Programme PT2020 for financial support to CIMO (UIDB/00690/2020). Dr. Gonzales-Barron acknowledges the national funding by FCT, P.I., through the Institutional Scientific Employment Programme contract. José M. Lorenzo is member of the HealthyMeat network, funded by CYTED (ref. 119RT0568). info:eu-repo/semantics/publishedVersion

Hepatitis A virus (HAV) is responsible for common acute viral hepatitis worldwide. Improvement in sanitation and use of efficient vaccines have reduced HAV incidence in developed countries. However, naive adult population are most susceptible to severe outcomes, and high endemic areas persist in developing regions. The transmission of HAV through the fecal-oral route is established. However, considering evolving consumption habits and global market exchange of food, investigations on risk factors associated with HAV infection are needed. Thus, a systematic review and a meta-analysis of case-control, cohort and transversal studies was performed to determine the main risk factors associated with sporadic HAV infection. Relevant scientific articles were identified through systematic literature search and subjected to a methodological quality assessment. Mixed-effects meta-analyses models were adjusted by population type to appropriate data partitions. HAV infections are defined by serological testing. The quality assessment stage selected 78 studies investigating risk factors for sporadic infections with hepatitis A conducted between 1985 and 2013. This meta-analysis confirmed that HAV infections are mostly related to inter-human transmissions, either due to contact with an ill person, through oral-anal sex practice, or lack of personal hygiene. Travel to endemic countries, occupational exposure such as working in child daycare, and exposure to wastewater were associated with HAV infection. As HAV can persist in the environment, it was not surprising that consumption of untreated drinking water, shellfish consumed raw, and crop products were risk factors. Food contamination could be due to the use of contaminated water (fruits, vegetables) or originate from infected food handlers at every point of the food chain (from picking to serving). Eating or drinking outside were associated with HAV infection. A lack of recent case-control studies was identified, with only three studies eligible between 2011 and 2017. Case-control studies required a more precise definition of risk factors such as type of crop product, and storing/preparation information (e.g. washed, frozen). The frequency of consumption or duration of environmental exposure could also better inform relationship between exposure and risk of infection. In a context of epidemiological change of HAV, international travel and trade of foods, future case-control studies are needed and should focus on populations at risk of severe infections and acute cases. U. Gonzales-Barron and V. Cadavez are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under programme PT2020 for the financial support to CIMO (UID/AGR/00690/2019). info:eu-repo/semantics/publishedVersion

In order to design effective public health strategies, and, in particular, effective food safety interventions to reduce the burden of foodborne disease, the most important sources of enteric illnesses should be identified. Both case-control and cohort observational studies have for long been powerful approaches among epidemiologists to investigate the association of exposure and illness. In the literature, there are numerous case-control and cohort studies reporting results on risk factors and routes of transmission of sporadic foodborne infections. The objective of this article is to describe, in depth, the strategies implemented for systematic review and meta-analysis of the associations between multiple risk factors and eleven food and waterborne diseases, namely, non-typhoidal salmonellosis, campylobacteriosis, Shiga-toxin E. coli infection, listeriosis, yersiniosis, toxoplasmosis, norovirus infection, hepatitis A, hepatitis E, cryptosporidiosis and giardiasis. First, this article describes the procedures of systematic searches in five bibliographic engines, screening of relevance and assessment of methodological quality according to pre-set criteria. It proceeds with the explanation of a broad data categorisation scheme established to hierarchically group the risk factors into travel, host-specific factors and pathways of exposure (i.e., person-to-person, animal, environment and food routes), with views to harmonising and supporting the integration of outcomes from studies investigating a variety of potential determinants of disease. Next, the article describes the four meta-analysis models that were devised in order to calculate: (i) overall oddsratios of acquiring the disease due to a specific risk factor by geographical region; (ii) overall odds-ratios of acquiring the disease from the different risk factors; (iii) overall risks of disease from consumption of ready-toeat and barbecued foods; and (iv) overall effects of food handling (i.e., consuming food in raw, undercooked or unwashed state) and food preparation setting (i.e., eating food prepared outside the home) on risk of disease. The procedures for sensitivity analysis and removal of any influential and potentially-biased odds-ratio; and two methods for publication bias assessment are outlined. Finally, details are given on deviations from the standard risk categorisation scheme for specific foodborne hazards. info:eu-repo/semantics/publishedVersion

This study aims to evaluate the effects of camu-camu powder (CCP), Amazonian berry fruit with documented bioactive properties, physicochemical meat parameters, and the growth kinetics parameters of S. enterica ser. Typhimurium, psychrotrophic bacteria (PSY), and lactic acid bacteria (LAB) in vacuum-packed ground beef. Batches of ground beef were mixed with 0.0%, 2.0%, 3.5%, and 5.0% CCP (w/w), vacuum-packed as 10 g portions, and stored at 5 °C for 16 days. Centesimal composition analyses (only on the initial day), pH, TBARS, and color were quantified on storage days 1, 7, and 15, while PSY and LAB were counted on days 0, 3, 6, 9, 13, and 16. Another experiment was conducted with the same camu-camu doses by inoculating S. enterica ser. Typhimurium microbial kinetic curves were modeled by the Huang growth and Weibull decay models. CCP decreased TBARS in beef from 0.477 to 0.189 mg MDA·kg−1. No significant differences in meat pH between treated and control samples were observed on day 15. CCP addition caused color changes, with color a* value decreases (from 14.45 to 13.44) and color b* value increases (from 17.41 to 21.25), while color L* was not affected. Higher CCP doses caused progressive LAB growth inhibition from 0.596 to 0.349 log CFU·day−1 at 2.0% and 5.0% CCP, respectively. Similarly, PSY growth rates in the treated group were lower (0.79–0.91 log CFU·day−1) compared to the control (1.21 log CFU·day−1). CCP addition at any of the investigated doses produced a steeper S. enterica ser. Typhimurium inactivation during the first cold storage day, represented by Weibull’s concavity α shape parameter, ranged from 0.37 to 0.51, in contrast to 1.24 for the control. At the end of the experiment, however, S. enterica ser. Typhimurium counts in beef containing CCP were not significantly different (p &lt, 0.05) from the control. Although CCP affects bacterial kinetics, it does not protect ground beef against spoilage bacteria and Salmonella to the same degree it does against lipid peroxidation.

Health concerns have led to a search for natural plant-based antimicrobials. Ellagic acid has been shown to have antimicrobial activity against foodborne pathogens. The objective of this study was to assess the effect of a high-ellagic acid commercial pomegranate on the heat resistance of Escherichia coli O104:H4 in ground chicken. A full 2 4 factorial design was used, consisting of temperature treatment with four levels (55.0, 57.5, 60.0, and 62.5 °C) and pomegranate with four levels (0.0, 1.0, 2.0, and 3.0 wt/wt. % containing 70% ellagic acid). Experiments were conducted twice, providing a total of 32 survival curves. A three-parameter Weibull primary model was used to describe survival kinetics. Secondary models were then developed to estimate the shape parameter ß (i.e., curvature representing susceptibility of cells to stress), scale parameter ? (i.e., time to reach the first decimal reduction) and the 5.0-log lethality time t 5.0 (i.e., time to reach a 5.0-log reduction), all as polynomial functions of temperature and pomegranate powder concentration. The positive effect of pomegranate concentration on both ß and ? demonstrated that the phenolic-rich pomegranate powder causes E. coli O104:H4 cells to become more susceptible to heat, increasing the steepness and concavity of the isothermal survival curves. It was estimated that the 5.0-log reduction time would reach a minimum at a pomegranate powder concentration of 1%, producing a 50% decrease in lethality time, in comparison to that without added pomegranate powder. Nonetheless, a mixed-effect omnibus regression further confirmed that the greatest difference in the thermal resistance of E. coli O104:H4 happened between tests with and without pomegranate powder. In fact, adding more than 1.0% pomegranate powder, at a constant temperature, resulted only in a marginal decrease in thermal resistance. Meat processors can use the model to design lethality treatments in order to achieve specific reductions of E. coli O104:H4 in ground chicken. We thank Angie Osoria and Carol E. Levin for technical assistance as well as in facilitating the preparation of the manuscript. Also, Dr. Gonzales-Barron wishes to acknowledge the financial support provided by the Portuguese Foundation for Science and Technology (FCT) through the award of a five-year Investigator Fellowship (IF) in the mode of Development Grants (IF/00570). info:eu-repo/semantics/publishedVersion

Complete microbiological, physicochemical and environmental longitudinal data were obtained from factory surveys in order to elucidate risk factors leading to the survival of Enterobacteriaceae, Staphylococcus aureus and Listeria monocytogenes in salpicão sausage. A total of six batches were surveyed from two factories; one added nitrite and polyphosphates in their formulation (Factory II). The addition of polyphosphates slowed down fermentation, which favoured the increase in Enterobacteriaceae (∼0.5 log CFU/g), S. aureus (∼0.5 log CFU/g) and L. monocytogenes (∼70 CFU/g) until maceration. Maceration should be regarded as a critical stage in the control of S. aureus in salpicão, as longer maceration (p = 0.033), higher temperatures of maceration (p = 0.018) and pH of macerated meat (p = 0.079) led to higher S. aureus counts in the final product. The main hurdle controlling S. aureus in salpicão is the pH. For L. monocytogenes and Enterobacteriaceae, at least three hurdles hinder their viability: low Aw (p = 0.010; 0.012), smoking (p

The objective of this study was to model the fate of Listeria monocytogenes inoculated in beef at two concentrations (2.5 and 4.0 log CFU/g), packaged under air, vacuum and three modified atmospheres MAP: 70%O 2 /20%CO 2 /10%N 2 , 50%O 2 /40%CO 2 /10%N 2 and 30%O 2 /60%CO 2 /10%N 2 , and refrigerated at a normal temperature (4 °C) and at a mild abusive temperature (9 °C). The experimental design produced a total of 20 environmental conditions. An omnibus model based on the Weibull equation proved statistically that L. monocytogenes survives better in vacuum (VP) than in aerobic conditions, although without significant difference in its ability to survive in the temperature range between 4 °C and 9 °C. Furthermore, regardless of the refrigeration temperature, the presence of CO 2 in the package atmosphere exerted a bactericidal effect on L. monocytogenes cells, being approximately 1.5 log of reduction when storage time reached 10 days. Since the pathogen can survive in VP/MAP beef, there is a need of maintaining its numbers below 100 CFU/g before packaging by placing efforts on the implementation of control measures during processing. The authors would like to thank CECAV-UTAD and the research is supported by national funds by FCT- Portuguese Foundation for Science and Technology, under the PEst-OE/AGR/UI0772/2014. Dr. Gonzales–Barron wishes to acknowledge the financial support provided by the Portuguese Foundation for Science and Technology (FCT) through the award of a five-year Investigator Fellowship (IF) in the mode of Development Grants (IF/00570). info:eu-repo/semantics/publishedVersion

The fate of Listeria monocytogenes during ripening of artisanal Minas semi-hard cheese, as influenced by cheese intrinsic properties and by autochthonous (naturally present) or intentionally-added anti-listerial lactic acid bacteria (LAB) was modeled. Selected LAB strains with anti-listerial capacity were added or not to raw or pasteurized milk to prepare 4 cheese treatments. Counts of LAB and L. monocytogenes, pH, temperature and water activity were determined throughout cheese ripening (22 days, 22±1ᵒC). Different approaches were adopted to model the effect of LAB on L. monocytogenes: an independent approach using the Huang primary model to describe LAB growth and the linear decay model to describe pathogen inactivation; the Huang-Cardinal [pH] model using the effect of pH variation in a dynamic tertiary approach; and the Jameson-effect with Nmax tot model which simultaneously describes L. monocytogenes and LAB fate. L. monocytogenes inactivation occurred in both treatments with added LAB and inactivation was faster in raw milk cheese (−0.0260 h−1) vs. pasteurized milk cheese (−0.0182 h−1), as estimated by the linear decay model. Better goodness-of-fit was achieved for the cheeses without added LAB when the Huang primary model was used. A faster and great pH decline was detected for cheeses with added LAB, and the Huang-Cardinal [pH] model predicted higher pathogen growth rate in cheese produced with raw milk, but greater L. monocytogenes final concentration in pasteurized milk cheese. The Jameson-effect model with Nmax tot predicted that LAB suppressed pathogen growth in all treatments, except in the treatment with pasteurized milk and no LAB addition. The Huang-Cardinal [pH] model was more accurate in modeling L. monocytogenes kinetics as a function of pH changes than was the Jameson-effect model with Nmax tot as a function of LAB inhibitory effect based on the goodness-of-fit measures. The Jameson-effect model may however be a better competition model since it can more easily represent L. monocytogenes growth and death. This study presents crucial kinetic data on L. monocytogenes behavior in the presence of competing microbiota in Minas semi-hard cheese under dynamic conditions. This work was supported by The São Paulo Research Foundation – FAPESP [grant numbers 2014/14891–7 and 2016/09346–5]; CNPq - The National Council for Scientific and Technological Development [grant numbers 302763/2014–7 and 305804/2017–0]; and CAPES –Coordination for the Improvement of Higher Education Personnel [Finance Code 001]. info:eu-repo/semantics/publishedVersion

Previous research showed that meat of optimal tenderness is produced when rigor mortis temperature falls between 12−35 °C. This study aimed to classify beef carcasses quality according to the ideal window rule using pH/temperature decay descriptors and animal characteristics. Seventy-four Mirandesa breed and 52 Crossbreds, with an average age of 10.1 ± 2.32 months, were slaughtered at one abattoir located in the Northeast of Portugal. Carcass temperature and pH, logged during 24 h post-mortem, were modelled by exponential decay equations that estimated temperature (kT) and pH (kpH) decay rates. Additionally, other pH/temperature descriptors were estimated from the fitted models. From linear models adjusted to each descriptor, it was found that hot carcass weight, age, breed, gender, age class, fat cover, conformation and transport and lairage time had influence (P < 0.05) on pH and temperature decay rates. Thus, combining the variables kT and kpH, and selected animal/carcass characteristics as linear predictors, a system to classify quality of carcasses was developed. The authors are grateful to the support from the’Associação de Criadores de Bovinos de Raça Mirandesa’ for kindly providing the meat samples and’Terra Fria Carnes' for putting their slaughterhouse facilities at our disposal during the experimental trials. info:eu-repo/semantics/publishedVersion

This study compares dynamic tertiary and competition models for L. monocytogenes growth as a function of intrinsic properties of a traditional Brazilian soft cheese and the inhibitory effect of lactic acid bacteria (LAB) during refrigerated storage. Cheeses were prepared from raw or pasteurized milk with or without the addition of selected LAB with known anti-listerial activity. Cheeses were analyzed for LAB and L. monocytogenes counts, pH and water activity (aw) throughout cold storage. Two approaches were used to describe the effect of LAB on L. monocytogenes: a Huang-Cardinal model that considers the effect of pH and aw variation in a dynamic kinetic analysis framework; and microbial competition models, including Lotka-Volterra and Jameson-effect variants, describing the simultaneous growth of L. monocytogenes and LAB. The Jameson-effect with γ and the Lotka-Volterra models produced models with statistically significant coefficients that characterized the inhibitory effect of selected LAB on L. monocytogenes in Minas fresh cheese. The Huang-Cardinal model [pH] outperformed both competition models. Taking aw change into account did not improve the fit quality of the Huang-Cardinal [pH] model. These models for Minas soft cheese should be valuable for future microbial risk assessments for this culturally important traditional cheese Authors are grateful to Fundação de Amparo à Pesquisa do Estado de São Paulo for the financial support (Grants 2014/14891-7 and 2016/ 09346-5). Dr. Gonzales-Barron acknowledges the financial support provided by the Portuguese Foundation for Science and Technology through the award of an Investigator Fellowship (IF) in the mode of Development Grants (IF/00570). A. S. Sant’Ana acknowledges Conselho Nacional de Desenvolvimento Científico e Tecnológico (Grant # 302763/2014-7, 305804/2017-0), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Finance Code 001”. info:eu-repo/semantics/publishedVersion

The objective of this study was to evaluate the combined effect of xanthan gum (XG) on physicochemical, rheological and textural properties of gluten-free batter and bread. To prepare gluten-free batter, different levels of XG (1.5, 2.5 and 3.5%) and water (90, 100 and 110%) were added to a base formula of rice (50%), maize (30%) and quinoa flours (20%); and the batters were evaluated in a factorial design. Several properties on both batter (stickiness and back extrusion) and its corresponding bread (loaf specific volume, baking loss, water activity and pH, texture profile, mean cell area, mean cell density, cell size uniformity, void fraction, mean cell compactness and mean cell aspect ratio) were then evaluated. Higher XG doses (p < .001) tended to produce batters of lower stickiness, adhesion and cohesive-strength, yet, of higher firmness, consistency, cohesiveness and viscosity index. After baking, these loaves presented lower specific volume; lower crumb aw, pH, hardness, springiness, mean cell area and void fraction; and higher (p < .001) chewiness, resilience, mean cell density, cell size uniformity and mean cell compactness. The sticker and less consistent batters produced with higher WC rendered larger bread loaves of softer and more springy/resilient crumbs with greater mean cell size and void fraction. Gluten-free loaves of good appearance in terms of higher specific volume, lower crumb hardness, higher crumb springiness, and open grain visual texture were obtained in formulations with 110% WC and XG doses between 1.5 and 2.5%. Eng. Encina-Zelada acknowledges the financial aid provided by the Peruvian National Programme of Scholarships and Student Loans (PRONABEC) in the mode of PhD grants (Presidente de la República–183308). Dr. Gonzales-Barron wishes to acknowledge the financial support provided by the Portuguese Foundation for Science and Technology (FCT) through the award of a five-year Investigator Fellowship (IF) in the mode of Development Grants (IF/00570). The authors are grateful to Eng. Andrea Oliveira from Prodipani, Portugal, for kindly providing breadmaking ingredients. info:eu-repo/semantics/publishedVersion

The objective of this study was to investigate and summarize the levels of incidence of Salmonella spp., Listeria monocytogenes, Staphylococcus aureus and Campylobacter spp. in poultry meat commercialized in Europe. After systematic review, incidence data and study characteristics were extracted from 78 studies conducted in 21 European countries. Pooled prevalence values from 203 extracted observations were estimated from random-effects meta-analysis models adjusted by pathogen, poultry type, sampling stage, cold preservation type, meat cutting type and packaging status. The results suggest that S. aureus is the main pathogen detected in poultry meat (38.5%; 95% CI: 25.4–53.4), followed by Campylobacter spp. (33.3%; 95% CI: 22.3–46.4%), while L. monocytogenes and Salmonella spp. present lower prevalence (19.3%; 95% CI: 14.4–25.3% and 7.10%; 95% CI: 4.60–10.8%, respectively). Despite the differences in prevalence, all pathogens were found in chicken and other poultry meats, at both end-processing step and retail level, in packed and unpacked products and in several meat cutting types. Prevalence data on cold preservation products also revealed that chilling and freezing can reduce the proliferation of pathogens but might not be able to inactivate them. The results of this meta-analysis highlight that further risk management strategies are needed to reduce pathogen incidence in poultry meat throughout the entire food chain across Europe, in particular for S. aureus and Campylobacter spp.

The aim of this study was to evaluate and compare the quality characteristics, chemical composition and lipid profile of lamb meat from Italian (Biellese and Sambucana) and German (Texel-Merino-Blackhead-Charollais [TMBC]) breeds reared in extensive and semi-extensive production systems. Meat samples from 89 animals were analysed. The meat of the lambs from semi-intensively reared Biellese, and extensively reared Sambucana and TMBC breeds produced lean meat, with slightly higher intramuscular fat content in TMBC. The latter also produced meat of darker colour (P lt 0.05) and higher protein content (P lt 0.05). The meat of Sambucana lambs presented the lowest total cholesterol content (P lt 0.05). The fatty acid profile of the meat showed a clear advantage of both extensively reared breeds, which had substantially lower proportion of saturated but higher of polyunsaturated fatty acids, particularly n-3 (P lt 0.05). The beneficial effect of the extensive rearing conditions was associated with lower n-6/n3 ratio, and atherogenic and thrombogenic indices, thereby suggesting that production system can be used successfully to modify the fatty acid profile to achieve a positive effect for the human health. The authors are grateful to EU ERA-NET programme and the Portuguese Foundation for Science and Technology (FCT) for funding the project “EcoLamb–Holistic Production to Reduce the Ecological Footprint of Meat (SusAn/0002/2016). CIMO authors are grateful to FCT and FEDER under Programme PT2020 for financial support to CIMO (UIDB/00690/2020). Dr. Gonzales-Barron acknowledges the national funding by FCT, P.I., through the Institutional Scientific Employment Programme contract. Jos´e M. Lorenzo is member of the HealthyMeat network, funded by CYTED (ref. 119RT0568). info:eu-repo/semantics/publishedVersion

Yersinia enterocolotica is an important causative agent of diarrheal illness. A systematic review and meta-analysis of case-control studies were performed to determine the main risk factors associated with sporadic Y. enterocolitica infections. Suitable scientific articles published between 1987 and 2017 were identified through a systematic literature search and subject to methodological quality assessment. From each study, odds ratios (OR) were extracted or calculated, as well as study characteristics such as population type, design, type of model used and risk factor categorization. Mixed-effects meta-analytical models were adjusted by population type to appropriate data partitions. From 807 identified references, the quality assessment stage was passed by 14 case-control studies focusing on sporadic Y. enterocolitica infections which provided 165 ORs for meta-analysis. All studies considered Y. enterocolitica as the cause of sporadic infections and are mainly located in Europe. The meta-analysis identified host-specifics factors, animal and food exposures as significant risk factors. The meta-analysis confirms the predominant role of the pig reservoir. The occupational contact with pigs and the consumption of pork meat are significantly associated with sporadic Y. enterocolitica infections occurrence. The consumption of raw or undercooked pork meat is also a very important risk factor. Untreated drinking water was also identified as risk factor. Further studies with other enteropathogenic Yersinia species, especially Y. pseudotuberculosis, and/or from other continents would help to refine conclusions of the meta-analysis of the risk factors of yersiniosis. U. Gonzales-Barron and V. Cadavez are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020). U. Gonzales- Barron acknowledges FCT, P.I., for the institutional scientific employment programme. info:eu-repo/semantics/publishedVersion

Although sheep meat has a small share of 1.5 % of the total meat production in the EU, sheep farming is of great importance to rural development and the environment. Enhancing the quality of lamb meat of local breeds is essential to ensure both profitability for sheep producers and the conservation of endangered breeds. This study aimed to (i) characterise the evolution of spoilage microorganisms in refrigerated vacuum-packed lamb meat from a total of 10 farms housing 8 local breeds of Portuguese, Spanish, Italian and Slovenian origin raised in intensive, extensive or semi-extensive regime; and (ii) elucidate how intrinsic properties of meat can affect its microbial spoilage. Cold carcass weight (CCW), ultimate pH (pH24) and proximate analysis were quantified on carcass/meat from each of the 285 animals raised and slaughtered for this purpose; while mesophiles, lactic acid bacteria, Pseudomonas spp. and psychrotrophic bacteria were enumerated during 15-day storage at 4 °C. Substantial variability in all attributes were found between the ten farms. CCW of intensively-raised lambs (21.4 kg; 95 % CI: 20.6–22.1 kg) were higher (p lt 0.05) than the ones in semi-extensive regime (14.9 kg; 95 % CI: 14.4–15.4 kg), and in turn these were heavier (p lt 0.05) than the extensively raised lambs (12.4 kg; 95 % CI: 12.0–12.7). Mean contents of protein (76.5–87.4% db), fat (3.78–13.1% db) and ashes (4.62–5.65% db) in lamb meat were highly dependent on the farm. Although meat from some farms was associated to higher microbial levels, in general, microbial growth was found to be modulated by intrinsic properties of meat. Higher pH24 (p lt 0.05), moisture (p lt 0.05), protein content (p lt 0.05) and ashes content (p lt 0.01) accelerated spoilage rate; whereas meat from heavier carcasses (p lt 0.001) and of higher fat content (p lt 0.01) presented slower growth of spoilage bacteria. In order to improve the microbial quality of lamb meat, animal handling must be enhanced to minimise pre-slaughter stress; slaughtering practices and hygiene must be improved; and a carcass classification system could be adopted towards the selection of fatter animals and chilled carcasses of optimal pH24. The authors are grateful to EU ERA-NET programme and the Portuguese Foundation for Science and Technology (FCT) for funding the project “EcoLamb–Holistic Production to Reduce the Ecological Footprint of Meat (SusAn/0002/2016). CIMO authors are grateful to FCT and FEDER under Programme PT2020 for financial support to CIMO (UIDB/00690/2020). Dr. Gonzales-Barron acknowledges the national funding by FCT, P.I., through the Institutional Scientific Employment Programme contract. José M. Lorenzo is member of the HealthyMeat network, funded by CYTED (ref. 119RT0568). The authors would like to acknowledge also Elena Saccà and Angela Sepulcri for providing technical support in laboratory analyses. info:eu-repo/semantics/publishedVersion

Toxoplasmosis is considered as the most prevalent parasitic zoonotic infection worldwide. The parasitic cycle is mostly known, but the relative contribution of different sources and pathways of transmission was not previously studied by a meta-analysis. A systematic review and a meta-analysis of case-control, cohort, and cross-sectional studies were performed to determine the main risk factors associated with sporadic Toxoplasma infection. Suitable scientific articles were identified through a systematic literature search and subjected to a methodological quality assessment. Mixed-effects meta-analysis models were adjusted by population type – children, mixed population, and pregnant women – to appropriate data partitions. 187 primary studies passed the quality assessment stage, investigating risk factors for sporadic infection with Toxoplasma gondii conducted between 1983 and 2016. Cases were defined by serology. The meta-analysis of Toxoplasma sporadic infections revealed the significance of transmission by environmental factors such as contact with soil and contact with animals, in particular cats. The consumption of raw or undercooked meat and unwashed vegetables significantly increased the odds of acquiring the disease. Shellfish and raw milk were identified as significant sources of toxoplasmosis. Almost all meat categories were identified as risk factors: pork, poultry, beef, processed meat, lamb, and game meat. Contaminated drinking water may play a role in the acquisition of infection. Moreover, the lack of hygiene in preparing food was identified as a risk factor. A significant risk factor for pregnant women is traveling abroad. Lastly, blood transfusion (in pregnant women) and immunocompromised conditions were found associated with positive serology. The broad definition of exposures and the use of serology for the case definition are the main limitations for the interpretation of the results of this meta-analysis. The transmission pathways require further investigations using longitudinal studies and subtyping approaches. The authors would like to thank ANSES staff and the members of the ANSES Working Group on Source Attribution of Foodborne Diseases: Moez Sanaa, Laurence Watier, Jean Christophe Augustin, Fr´ed´eric Carlin, Julie David, Philippe Fravalo, Laurent Guillier, Nathalie Jourdan-Da Silva, Alexandre Leclercq, Lapo Mughini-Gras, Nicole Pavio. U. Gonzales-Barron and V. Cadavez are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for the financial support to CIMO (UID/AGR/00690/2019). U. Gonzales-Barron acknowledges the national funding by FCT, P.I., through the Institutional Scientific Employment Program contract. info:eu-repo/semantics/publishedVersion

Non-typhoidal Salmonella is an important causative agent of diarrheal illness worldwide. A systematic review and meta-analysis of case-control studies were performed to determine the main risk factors associated with sporadic salmonellosis. Suitable scientific articles published up to 2017 were identified through a systematic literature search and subject to methodological quality assessment. From each study, odds ratios (OR) were extracted or calculated, as well as study characteristics such as population type, design, type of model used and risk factor categorization. Mixed-effects meta-analytical models were adjusted by population type to appropriate data partitions. From 3858 identified references, the quality assessment stage was passed by 62 case-control studies focusing on sporadic salmonellosis which provided 1154 ORs for meta-analysis. The meta-analysis identified host-specifics factors, travel, environmental, animal and food exposures as significant risk factors of salmonellosis in the mixed population. For the mixed population, foods significantly associated with salmonellosis were eggs and egg products, composite foods, and meat (pork, red meats other than beef and poultry meats). In the child population, the main risk factors were found for person-to-person transmission, recent use of gastric anti-acids or antibiotics, contact with pets and farm animals, environmental, and food vehicles. Breastfeeding was found to be a protective factor for children. The food vehicles identified in children comprised dairy (milk formula), produce, meat and eggs products. Untreated drinking water was a risk factor for the children and mixed population. The result of the meta-analysis conducted at the international level is very important in the context of increasing international trade in foodstuffs and changes in food consumption patterns. The authors would like to thank Anses staff and the members of the Anses Working Group on Source Attribution of Foodborne Diseases: Moez Sanaa, Jean Christophe Augustin, Fr´ed´eric Carlin, Alexandre Leclercq, Nicole Pavio, Isabelle Villena, and Laurence Watier. U. Gonzales-Barron and V. Cadavez are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020). U. Gonzales- Barron acknowledges the national funding by FCT, P.I., through the institutional scientific employment program contract. info:eu-repo/semantics/publishedVersion

Norovirus is responsible for 20% of acute gastroenteritis worldwide. The fecal-oral route of transmission is known, but we proposed a first attempt to identify the relative importance of different sources and vehicles for sporadic cases using meta-analysis models. Case-control and cohort/cross-sectional studies were systematically reviewed and analyzed to assess the main risk factors associated with sporadic norovirus infections. Suitable scientific articles were identified through systematic literature search and subjected to a methodological quality assessment. Mixed-effects meta-analyses models were adjusted by population type to appropriate risk factor categories. The quality assessment stage led to include 14 primary studies conducted between 1993 and 2014. From these, eight studies investigated exposures in children/infants, and eight concerned the mixed population. The meta-analysis confirmed the oro-fecal route for norovirus infections, with the person-to-person transmission (pooled OR=3.002; 95% CI: [2.502 -3.060] in mixed population), and the lack of personal hygiene (pooled OR=2.329; 95% CI: [1.048 -5.169]). The meta-analysis also enlightened the role of indirect transmission through the environment with pathways like untreated drinking water (mixed population), with a pooled OR=2.680 (95% CI: [1.081-6.643]) and farm environment (children population). Indirect transmission also involved the food pathway, which was finally found significant with consumption of seafood (mixed population) (pooled OR=2.270; 95% CI: [1.299-3.968]) and composite food (eating outside/uncooked mixed and young population) (pooled OR=4.541; 95% CI: [3.461-5.958]). These results are coherent with the findings from studies on outbreaks. However, a too broad definition of exposure factors limited the interpretation of results, as occurred with the seafood pathways that combined fish and shellfish. Other factors such as consumption of Food-handled products or the type of drinking water deserveE to be better investigated. Furthermore, better harmonization in case definition and appropriate case-control or cross-sectional studies would allow better addressing sporadic cases risk factors, especially for susceptible populations, such as children, elderly or immunosuppressed persons. U. Gonzales-Barron and V. Cadavez are grateful to the Foundation for Food Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/ 2019). U. Gonzales-Barron thanks the national funding by FCT, P.I., through the Institutional Scientific Employment Program contract. info:eu-repo/semantics/publishedVersion

The objective of this study was to investigate, by means of a D-optimal mixture design, the combined effects of hydroxypropyl methyl cellulose (HPMC), xanthan (XG), and guar (GG) gums on physicochemical, rheological, and textural properties of gluten-free batter and bread. For each of the quality properties measured, a two-factor interaction model was fitted, and the significance of its terms was assessed by analysis of variance. Sticky batters were produced with a combination of high dose of GG (0.60%), high-intermediate dose of HPMC (3.36%), and low dose of XG (0.04%). Combinations of high XG dose (0.60%) and intermediate doses of HPMC (3.08%) and GG (0.32%) rendered GF breads of greater specific volume, while lower bread crust luminosity was obtained with combinations of high GG dose (0.60%), low XG dose (0.04%), and high-intermediate HPMC dose (3.36%). Combinations of high-intermediate HPMC dose (3.36%), high GG dose (0.60%), and low XG dose (0.04%) produced both softer crumbs and bread slices of more open visual texture. By using a desirability function that maximized specific volume while minimizing crust luminosity, crumb hardness, and mean cell density, the optimization of hydrocolloids mixture rendered a value of 0.54, for a combination of 0.24% XG, 0.60% GG, and 3.16% HPMC., Encina-Zelada acknowledges the financial aid provided by the Peruvian National Program of Scholarships and Student Loans (PRONABEC) in the mode of PhD grants (Presidente de la República-183308)., info:eu-repo/semantics/publishedVersion

Biopreservatives such as plant-based antimicrobials and bacteriocinogenic starter cultures have been proposed as hurdles to increase microbiological safety of a variety of products, including cheese, and numerous studies have reported their pathogen inhibitory properties. For that reason, the objective of this meta-analysis was to summarise the inactivation of Listeria monocytogenes (LM), Staphylococcus aureus (SA) and Salmonella spp. (SS) in cheese attained by added lactic acid bacteria (LAB) and essential oils (EOs); and to compare the inhibitory effectiveness by application mode and specific antimicrobial. After systematic review, 1810 observations on log reduction data and study characteristics were extracted from 53 studies. Comparing among the factual methods of application of antimicrobials (in milk, cheese surface and incorporated in films), meta-regression models pointed out that addition of EOs to milk renders, as a whole, the lowest inhibitory effect against LM, SA and SS in the finished product; whereas for added LAB, incorporation in milk prompts a faster inactivation of LM than onto cheese surface. Lemon balm, sage and basil EOs showed the best inhibitory outcomes against LM and SA; whereas clove, oregano and bay EOs presented the highest bactericidal effect against SS. For a given increase in EO concentration, the application on cheese surface provides the greatest inhibitory effect against LM and SS, while EO-embedded films lead to a more rapid inactivation during maturation/storage. The experimental practice of inoculating the antimicrobial in cheese mixture should no longer be employed in challenge studies, since the meta-regression models have demonstrated that this application method biases the results, overestimating or underestimating the inhibitory effects of EOs or added LAB, respectively. This meta-analysis has also emphasised the need to further investigate the relationship between pathogen's inoculum size and their concentrations in time., The authors are grateful to EU PRIMA programme and the Portuguese Foundation for Science and Technology (FCT) for funding the ArtiSane Food project (PRIMA/0001/2018). This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit and Bio TecNorte operation (NORTE-01–0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. Ms. Silva wishes to acknowledge the financial support provided by FCT through the PhD grant SFRH/BD/137801/2018. The authors are grateful to FCT and FEDER under Programme PT2020 for financial support to CIMO (UIDB/00690/2020). Dr. Gonzales-Barron acknowledges the national funding by FCT, P.I., through the Institutional Scientific Employment Programme contract., info:eu-repo/semantics/publishedVersion

The objective of this study was to evaluate the effect of partially replacing two types of wheat flour (low ash content [type 55] and high ash content [type 65]) with Peruvian Prosopis pallida (mesquite) pod flour (0, 5, 10, 15%) on the nutritional quality and staling of composite breads. Mesquite flour (MF) enhanced the nutritional quality by increasing the fibre contents and unsaturated fatty acids of the bread. MF did not affect crumb hardness either when prepared with wheat flour type 65 (p = 0.374) or 55 (p = 0.122), but reduced crumb resilience (p 0.001) and water activity (p = 0.003) in both wheat flour types. When blended with wheat flour type 55, increasing levels of MF delayed the dehydration (p 0.001) and resilience loss rates. Likewise, the higher the MF level, the slower the crumb hardening of composite breads formulated with wheat flour type 55 (p = 0.028). Thus, MF did not only enhance the nutritional profile of composite breads, but could also retard staling as a supplement of wheat flour type 55. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020); L. Barros, U. Gonzales-Barron and M. Carocho also thank the national funding by FCT, P.I., through the institutional and individual scientific employment program-contract for their contracts. The authors are also grateful to FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E and TRANSCoLAB 0612_TRANS_CO_LAB_2_P. info:eu-repo/semantics/publishedVersion

The objective of this study was to assess the combined effect of guar gum (GG) and water content (WC) on the rheological properties of batter, and the physicochemical and textural properties of bread. Batches of gluten-free bread used a base formulation of rice (50%), maize (30%) and quinoa flour (20%), with different levels of GG (2.5, 3.0 or 3.5%) and water (90, 100 or 110%) in a full factorial design. Higher GG doses (p, Eng. Encina-Zelada acknowledges the financial aid provided by the Peruvian National Programme of Scholarships and Student Loans (PRONABEC) in the mode of PhD grants (Presidente de la República-183308). The authors are grateful to Eng. Andrea Oliveira from Prodipani, Portugal, for her kind advice and providing breadmaking ingredients., info:eu-repo/semantics/publishedVersion

Cryptosporidium spp. is an important cause of gastrointestinal disease worldwide, responsible for 69 million cases of illness in 2016. Information on the sources and transmission pathways of human cryptosporidiosis results mainly from outbreak investigations. A systematic review and a meta-analysis of case-control and cohort studies were performed to determine the main risk factors associated with sporadic cryptosporidiosis. Suitable scientific articles were identified through a systematic literature search and subjected to a methodological quality assessment. From each study, odds ratio (OR) measures were extracted or calculated, as well as study characteristics such as population type, design, type of model and risk factor hierarchy. Mixed-effects meta-analysis models were adjusted by population type to appropriate data partitions. From 1985 identified references, the quality assessment stage was passed by 57 cohort and case-control studies focusing on sporadic cryptosporidiosis. The eligible studies were conducted between 1983 and 2016 and provided 568 OR categorized for meta-analysis. This meta-analysis identified travel, immunocompromising conditions, contact with infected humans, waterborne transmission (contact with recreational waters, wastewater, and consumption of untreated drinking water), contact with animals and food consumption as the relevant risk factors for sporadic cryptosporidiosis. With regards to food exposures, consumption of meat, dairy products (raw milk) and dishes consumed outside home were found significantly associated with cryptosporidiosis. The consumption of poorly washed fruits and vegetables significantly increases ORs. This meta-analysis reveals that some potential sources of Cryptosporidium such as shellfish or vegetables are under-investigated. Future case-control studies for sporadic cryptosporidiosis should include population at risk, and investigate other potential sources in relation to the genotype and the subtype of Cryptosporidium spp. The authors would like to thank ANSES staff and the members of the ANSES Working Group on Source Attribution of Foodborne Diseases: Moez Sanaa, Laurence Watier, Jean Christophe Augustin, Frédéric Carlin, Julie David, Philippe Fravalo, Laurent Guillier, Nathalie Jourdan-Da Silva, Alexandre Leclercq, Lapo Mughini-Gras, Nicole Pavio. Ursula Gonzales-Barron and Vasco Cadavez are grateful to the Foundation for Food Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO UID/AGR/00690/2019. info:eu-repo/semantics/publishedVersion

This study estimated the risk of listeriosis from Brazilian cheese consumption using quantitative microbial risk assessment (QMRA). Risks associated to consumption of two cheese types were assessed: artisanal ripened semihard cheese (produced with raw milk) and refrigerated fresh soft cheese (produced with pasteurized milk). The semi-hard cheese model predicted Listeria monocytogenes growth or decline during ripening, while the soft cheese model predicted pathogen growth during refrigerated storage. Semi-hard cheese modeling scenarios considered L. monocytogenes starting concentration from −2.4 to 6 log CFU/mL in raw milk and three ripening times (4, 22 and 60 days). Soft cheese modeling scenarios considered L. monocytogenes starting concentration from −2.4 to 4 log CFU/mL in milk. The inclusion of anti-listerial lactic acid bacteria (LAB) in cheeses was also examined. Risk of listeriosis due to consumption of soft cheese was 6000 and 190 times greater than that of semi-hard cheese, for general and vulnerable populations, respectively. Aging semi-hard cheese reduced risk, and risk was influenced by L. monocytogenes starting concentration. Aging cheese with inhibitory LAB for 22 days reduced risk over 4 million-fold when L. monocytogenes was assumed to be 6 log CFU/mL in raw milk. The inclusion of inhibitory LAB also reduced risk of listeriosis due to soft cheese consumption, but not as much as for semi-hard cheese. QMRA results predicted that consumption of contaminated cheeses can carry a high risk of listeriosis, especially for vulnerable populations. Scenario analyses indicated that aging of semi-hard cheese and inclusion of antimicrobial LAB mix in semi-hard and soft cheeses are effective risk mitigation measures. The authors thank to Fundação de Amparo à Pesquisa do Estado de São Paulo for the financial support (Grants 2014/14891-7 and 2016/ 09346-5) and to Conselho Nacional de Desenvolvimento Científico e Tecnológico (Grant # 302763/2014-7 and #305804/2017-0), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Grant #33003017027P1). Dr. Gonzales-Barron wishes to acknowledge the financial support provided by the Portuguese Foundation for Science and Technology (FCT) through the award of a five-year Investigator Fellowship (IF) in the mode of Development Grants (IF/00570). info:eu-repo/semantics/publishedVersion

The increase of soluble solids in milk by the addition of powdered products is a normal practice in the elaboration of yogurt in order to enhance viscosity, texture and sensory properties for the consumer. The purpose of this study was to evaluate the effect of adding jumbo squid (Dosidicus gigas) powder (protein content, 420 g/kg) on the aforementioned quality properties of yogurt. Six formulations of yogurt were prepared with the addition of 0, 1, 3, 5, 7 or 10 g/100 mL jumbo squid powder, plus one formulation with 3 g/100 mL maltodextrin. During fermentation, yogurts formulated with squid powder gradually achieved greater viscosity while producing both more acidity and at a higher rate than the controls. Apart from lower pH (hence, higher titratable acidity) and higher viscosity, the final enriched yogurts presented lower syneresis than the controls (p < 0.05). Although the highest viscosity (58.90 Pa.s) and the lowest syneresis (1.00%) was achieved by the yogurt with 10 g/100 mL jumbo squid powder, it was the treatment formulated with 3 g/100 mL (pH 4.31, acidity 0.85 g/100 g, viscosity 40.90 Pa.s and syneresis 9.10%) that kept constant sensory properties, as evaluated by the panelists, while improving some physical properties of the control yogurts. The authors are grateful to Biologist Armando Solari and the Technological Institute of Production (ITP, Peru) for all support provided in the conduction of this research. Dr. Gonzales-Barron wishes to acknowledge the financial support provided by the Portuguese Foundation for Science and Technology (FCT) through the award of a five-year Investigator Fellowship (IF) in the mode of Development Grants (IF/00570). info:eu-repo/semantics/publishedVersion

Contamination of raw poultry meat with foodborne pathogens could occur because of improper handling at primary production and slaughterhouse levels. Low microbial prevalence data often consists of a high amount of non-detections (zero positives), so a flexible framework is required to characterise the underlying microbial distribution and conduct reliable inferential statistics. Thus, the objective of this work was to evaluate the performance of zero-inflated binomial (ZIB) regression models to describe the effects of sampling site (carcass, thigh, breast, wings) on the measured incidences of Salmonella, Listeria monocytogenes and Staphylococcus aureus on chicken meat. For each pathogen, four regression models based on the zero-inflated binomial ZIB (p, w0) distribution were fitted to the presence/absence data with sampling site as covariate and random-effects due to sampling occasion either in the binomial probability(p)or in the extra-proportion of non-detections(w0).Forthe three pathogens, the sampling site exerted a greater effect on w0 than on p itself, with breast bearing the lowest prevalence estimatesofSalmonellaspp.(mean: 0.88%;95%CI:0.02–1.95%)andS.aureus(mean1.48%;95%CI: 0.01–4.00%). The fitting capacity of the models was further improved when random effects due to sampling occasion were placed in w0 (deviances decreased from 146.7–156.7 to 140.2–140.6). This would imply that, theoretically, the variability in pathogens’ occurrence from batch to batch mainly arises from the variability in non-contaminated zones. At any sampling site, the mean prevalence was estimated as 1.35 (95% CI: 0.15 – 2.70) for Salmonella, 2.11 (95% CI: 0.04 – 5.63) for L. monocytogenes and 2.36 (95% CI: 0.04 – 5.12) for S. aureus. Sampling performance analysis showed that wings were mostly suitable to detect Salmonella and S. aureus with higher probability (0.016 and 0.035 respectively), while for L. monocytogenes, sampling of thigh could be more effective (0.032). Dr. Gonzales-Barron wishes to acknowledge the financial support provided by the Portuguese Foundation for Science and Technology (FCT) through the award of a five-year Investigator Fellowship (IF) in the mode of Development Grants ( IF/00570 ) info:eu-repo/semantics/publishedVersion

Artisanal raw milk cheeses are highly appreciated dairy products in Brazil and ensuring their microbiological safety has been a great need. This study reports the isolation and characterization of lactic acid bacteria (LAB) strains with anti-listerial activity, and their effects on Listeria monocytogenes during refrigerated shelf-life of soft Minas cheese and ripening of semi-hard Minas cheese. LAB strains (n ¼ 891) isolated from Minas artisanal cheeses (n ¼ 244) were assessed for anti-listerial activity by deferred antagonism assay at 37 C and 7 C. The treatments comprised the production of soft or semi-hard Minas cheeses using raw or pasteurized milk, and including the addition of selected LAB only [Lactobacillus brevis 2-392, Lactobacillus plantarum 1-399 and 4 Enterococcus faecalis (1-37, 2-49, 2-388 and 1-400)], L. monocytogenes only, selected LAB co-inoculated with L. monocytogenes, or without any added cultures. At 37 C, 48.1% of LAB isolates showed anti-listerial capacity and 77.5% maintained activity at 7 C. Selected LAB strains presented a bacteriostatic effect on L. monocytogenes in soft cheese. L. monocytogenes was inactivated during the ripening of semi-hard cheeses by the mix of LAB added. Times to attain a 4 log-reduction of L. monocytogenes were 15 and 21 days for semi-hard cheeses produced with raw and pasteurized milk, respectively. LAB with anti-listerial activity isolated from artisanal Minas cheeses can comprise an additional barrier to L. monocytogenes growth during the refrigerated storage of soft cheese and help shorten the ripening period of semi-hard cheeses aged at ambient temperature. The authors thank the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) for financial support (Grants 13/ 20456-9, 14/14891-7, 15/25641-4, 16/21041-5, 16/12406-0 and 16/ 09346-5). Dr. Gonzales-Barron also acknowledges the financial support provided by the Portuguese Foundation for Science and Technology (FCT) through the award of an Investigator Fellowship (IF) in the mode of Development Grants (IF/00570). A.S. Sant'Ana acknowledges the financial support of “Conselho Nacional de Desenvolvimento Científico e Tecnológico” (CNPq) (Grant #302763/ 2014-7) and CAPES (Grant #33003017027P1). info:eu-repo/semantics/publishedVersion

Linguiça is a Portuguese traditional fermented sausage whose microbiological quality and safety can be highly variable. In order to elucidate risk factors and the particularities of the manufacturing technology that explain the between-batch variability in total viable counts (TVC), Enterobacteriaceae, Staphylococcus aureus and Listeria monocytogenes in the product; microbiological and physicochemical characterisation of linguiça at five stages of production (i.e., raw pork meat, mixed with ingredients, macerated, smoked and ripened) was carried out. A total of six production batches were surveyed from two factories; one utilised curing salts and polyphosphate in their formulation (Factory II). The delayed fermentation in the nitrite-formulated sausages was partly responsible for the increase (p < 0.01) in Enterobacteriaceae, S. aureus and L. monocytogenes from raw meat (3.21logCFU/g, 1.30logCFU/g and 22.2CFU/g, respectively) to the end of maceration (4.14logCFU/g, 2.10logCFU/g and 140CFU/g, respectively) while the better acidification process in the nitrite-free sausages (Factory I) led to lower counts of S. aureus (2.64logCFU/g) and L. monocytogenes (10CFU/g) in the finished products. In Factory II, although L. monocytogenes entered the chain at the point of mixing, it became steadily inactivated during smoking and ripening ( < 50CFU/g), despite the initially-delayed fermentation. Nitrite had a strong effect on reducing Enterobacteriaceae throughout smoking (r=-0.73) and ripening (r=-0.59), while it failed to control the growth of S. aureus. The main hurdle preventing the development of S. aureus in linguiça is the pH, and other factors contributing to its control are: longer ripening days (p=0.019), low S. aureus in raw meat (p=0.098), properly-washed casings (p=0.094), and less contamination during mixing (p=0.199). In the case of L. monocytogenes, at least three hurdles hinder its development in linguiça: low a w (p=0.004), low pH (p=0.040) and nitrite (p=0.060), and other factors contributing to its control are: longer ripening (p=0.072) and maceration (p=0.106) periods, lower a w at the end of smoking (p=0.076) and properly-washed casings (p=0.099). Results have shown that there is a need to standardise the productive process of linguiça, to optimise the initial acidification process, and to reinforce proper programmes of quality control of ingredients and good hygiene practices, so as to minimise the introduction of Enterobacteriaceae and pathogens from external sources. This research was supported through the project PTDC/AGR-TEC/3107/2012, awarded by the Portuguese Foundation for Science and Technology (FCT), European Regional Development Funds (ERDF). Dr. Gonzales-Barron also acknowledges the financial support provided by FCT through the award of a five-year Investig info:eu-repo/semantics/publishedVersion

A meta-analysis on the influence of the pasture on the lipid content and fatty acid profile in lamb meat was conducted. A total of 20 studies were selected and data on total lipid content, saturated, mono- and polyunsaturated fatty acids in lamb meat were extracted for two population groups, indoors, and pastured. Due to the high between-study heterogeneity, separate random-effects models were applied to the raw mean difference (effect size parameterization) for each of the outcomes. The results of the meta-analysis pointed that access to pasture tended to decrease the fat content in lamb, while increasing the saturated fatty acids (P < 0.05). The amounts of monounsaturated and n-6 polyunsaturated fatty acids were decreased (P < 0.05). On the other hand, grazing led to substantial increase in the content of n-3 polyunsaturated fatty acids, and decrease (P < 0.01) of the ratio n-6/n-3 in lamb meat, thus suggesting that pasture rearing can be recommended for improvement of meat dietetic quality. by the Erasmus + Programme, Key Action 1: Learning mobility for individuals, administered by the European Union (№ 2014-1- BG01-KA103-000110). Dr. Gonzales-Barron wishes to acknowledge the financial support provided by the Portuguese Foundation for Science and Technology (FCT) through the award of five-year Investigator Fellowship (IF) in the mode of Development Grants (IF/00570). info:eu-repo/semantics/publishedVersion

In this study, a systematic review and meta-analysis were conducted to summarise available information on the occurrence of Salmonella spp. Listeria monocytogenes and shigatoxin-producing Escherichia coli (STEC) in fruits and vegetables sold at retail establishments in several European countries. Overall, L. monocytogenes was the main pathogen detected in all kinds of vegetables, packaged or not (3.4%; 95% CI: 2.15.4%) with Salmonella spp. being the pathogen of lowest incidence (0.9%; 95% CI: 0.51.2%). The pooled occurrence rate of pathogens in either packed or unpacked vegetables was estimated at 1.9% (95% CI: 1.23.1%), with 2.1% of prevalence (95% CI: 1.33.4%) for unpacked vegetables and 1.7% (95% CI: 0.92.9%) for packed ones. For the three pathogens, the category of spices and herbs was the most frequently contaminated with pathogens, whereas salads presented the lowest occurrence. The vegetable category with highest incidence of Salmonella spp. (1.7%; 95% CI: 0.74.1%) and L. monocytogenes (2.2%; 95% CI: 1.04.7%) is leafy greens whilst STEC is more frequently recovered from sprouts (1.9%; 95% CI: 0.55.9%). In the case of fruits, the pooled prevalence estimates for Salmonella spp., L. monocytogenes and STEC were 1.60% (0.54%; 95% CI: 0.554.60%), 1.91% (0.50%; 95% CI: 0.933.88%) and 4.71% (1.52%; 95% CI: 1.7312.2%), correspondingly., Dr. Gonzales-Barron wishes to acknowledge the financial support provided by the Portuguese Foundation for Science and Technology (FCT) through the award of a five-year Investigator Fellowship (IF) in the mode of Development Grants (IF/00570)., info:eu-repo/semantics/publishedVersion