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Queijo de Nisa PDO (Protected Designation of Origin) is a semi-hard cheese obtained from raw ewe's milk clotted with cardoon (Cynara cardunculus, L.) rennet. The aim of the present study was to characterize the bacterial communities naturally occurring in Queijo de Nisa PDO cheese samples through viable counting and metataxonomic analysis. Moreover, physico-chemical and morpho-textural analyses were also performed, together with the analysis of volatile organic compounds (VOCs) through solid-phase microextraction (SPME) coupled with gas chromatography-mass spectrometry (GC-MS). In the analyzed samples, pH values were comprised between 4.84 and 5.74. As for lactic acid, the concentration varied between 0.83 and 2.10 g 100 g -1 . Water activity varied between 0.942 and 0.960. Lightness of the samples ranged from 107.82 to 119.16, whereas hardness ranged between 34.45 N and 126.05 N. As for microbiological analyses, lactococci and lactobacilli counts were up to 9.01 Log CFU g -1 . Coagulase-negative cocci and enterococci counts were up to 7 Log CFU g -1 . Metataxonomic analysis revealed that Lactococcus lactis and Leuconostoc mesenteroides occurred at high frequency in all the analyzed samples. Moreover, Lactiplantibacillus plantarum, Lactococcus piscium, and Lacticaseibacillus zeae were also detected. The most represented VOCs were carboxylic acids, carbonyl compounds, alcohols, and esters. In the analyzed cheese samples, significant relationships between bacteria and VOCs were also evidenced. Of note, research on the microbiological and chemical features, as well as on morpho-textural characteristics, of Queijo de Nisa PDO  cheese can increase the knowledge on the interactions between raw materials, environment, and autochthonous microorganisms, thus enabling protection of biodiversity and tradition., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The aim of the present study was to characterize the bacterial and fungal communities naturally occurring in Queijo da Beira Baixa PDO cheese samples produced in Castelo Branco district (Beira Baixa Region, Portugal) through viable counts and metataxonomic analyses. Physico-chemical and morpho-textural analyses were also carried out, together with the analysis of volatile organic compounds (VOCs). In the analyzed samples, pH values ranged between 4.72 ± 0.15 and 5.85 ± 0.02, with values of lactic acid content comprised between 0.64 ± 0.00 and 1.95 ± 0.16 g 100 g -1 . Specific volume of cheese ranged from 1.09 ± 0.08 to 1.32 ± 0.02 g mL -1 . Texture profile analysis showed hardness ranging between 38.3 ± 9.6 N and 68.55 ± 7.5 N. As for lactic acid bacteria, presumptive lactococci, thermophilic cocci, and lactobacilli counts up to 9 Log cfu g -1 . Coagulase-negative cocci showed counts up to 7 Log cfu g -1 . Enterococci counts were up to 6 Log cfu g -1 . Finally, counts of eumycetes showed values up to 4 Log cfu g -1 . The results of metataxonomic analysis of bacteria showed the dominance of Lactococcus lactis in all the samples. Moreover, other taxa were detected, including Lactiplantibacillus plantarum, Loigolactobacillus coryniformis, Lactococcus piscium, Streptococcus thermophilus, and Lacticaseibacillus zeae. Mycobiota was characterized by the presence of Candida sake, Ustilago, Cladosporium variabile, Starmerella, Debaryomyces hansenii, and Pichia kluyveri. In the analyzed Queijo da Beira Baixa PDO cheese samples, carboxylic acids represented the most detected VOCs, followed by esters, carbonyl compounds, and alcohols., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

In the present study, bacterial and fungal diversity, as well as volatile profiles, of ready-to-eat Portuguese Painho de Porco Preto fermented sausages manufactured by two artisanal producers in the districts of Beja (producer A) and Evora (producer B) were studied. To this end, different selective growth media and a metataxonomic analysis were combined with Headspace Solid-Phase Microextraction-Gas Chromatography/Mass Spectrometry (HS-SPME-GC/MS) analysis. The results of the microbiological viable counts revealed active microbial populations of lactic acid bacteria (up to 8 Log cfu g -1 ), coagulase negative cocci (up to 6 Log cfu g -1 ), and eumycetes (up to 6 Log cfu g -1 ). Bacterial populations were characterized by high relative frequencies of Latilactobacillus sakei (up to 72%), together with Weissella and Staphylococcus equorum. The mycobiota was mainly dominated by Debaryomyces hansenii (up to 55% of the relative frequency) and Kurtzmaniella zeylanoides (up to 24% of the relative frequency). Unexpected species as Wickerhamomyces subpelliculosus and Zygosaccharomyces rouxii were also detected. HS-SPME-GC/MS analysis allowed to identify a complex volatile profile, showing 159 volatile organic compounds (VOCs). VOCs belonged to twelve classes, such as aldehydes, ketones and lactones, esters and acetates, alcohols, terpenoids, sulfur compounds, aliphatic hydrocarbons, aromatic hydrocarbons, nitrogen compounds, acids, furans and pyrans, and phenols. The analysis of VOCs composition provided evidence that samples from the two producers (A and B) were different, as confirmed by the Principal Component Analysis. Hence, it is likely that the raw materials used, as well as variations related with the empirical practice of the butchers, strongly influenced the final product. The results obtained in the present study represent a further advancement in the knowledge on the biodiversity and VOCs composition of Portuguese fermented sausages. To better understand the interactions occurring between the autochthonous microorganisms and the meat batter in the Painho de Porco Preto fermented sausage, microbial and VOCs dynamics must be further deepened throughout the production process., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The production of ovine or caprine milk cheeses with thistle rennet is a common practice in the Mediterranean basin. The aim of the present study was to obtain information on bacteria and yeast communities harboured by Queijo de Azeitão PDO cheese through viable counting and, for the first time, via metataxonomic analysis. Moreover, solid phase microextraction (SPME) technique was applied to characterize Queijo de Azeitão PDO cheese volatile compounds. Nine cheese samples were collected from three different artisan producers located in Portugal. The results of physico-chemical analyses showed significant differences between producers, with mean values ranging from 5.40 ± 0.25 (Producer 1) to 6.00 ± 0.22 (Producer 2). As for TTA, Producer 1 showed the highest mean value attesting at 18.04 ± 6.57 mL of 0.1 M NaOH used to reach pH 8.3. Regarding lactic acid concentration, Producer 1 showed the highest mean value attesting at 0.488 ± 0.106 g 100 g -1 , whereas, for acetic acid, no significant differences were evidenced among producers with values comprised between 0.141 ± 0.021 g 100 g -1 and 0.245 ± 0.016 g 100 g -1 . No significant differences were observed between overall mean values of the three producers for viable counts of presumptive lactococci, thermophilic cocci, presumptive lactobacilli, thermophilic lactobacilli and total mesophilic aerobes with values in the order of 7-8 log cfu g -1 . Moreover, no significant differences were evidenced for viable counts of coagulase-negative cocci, enterococci, Enterobacteriaceae and Pseudomonadaceae. As for eumycetes, cheeses from Producer 1 showed the lowest mean value (2.78 ± 2.42 log cfu g -1 ) in respect with values detected in cheeses from Producer 2 and 3. Concerning microbiota and mycobiota of the analyzed cheeses, the alpha diversity index did not show any significant difference between the three producers in terms of composition and complexity of the microbial population. A simple composition was apparently shared by the three producers, whose cheese manufactures were dominated by the presence of Leuconostoc mesenteroides (37% of the relative frequency in average), Lactococcus lactis (29%), Lacticaseibacillus zeae (4.7%), Lentilactobacillus kefiri (4.4%), Serratia spp. (3.5%), Lactiplantibacillus plantarum (2.7%), and Latilactobacillus sakei (2.5%). The mycobiota composition showed the neat dominance of Yarrowia lipolytica (46.7% of the relative frequency in average), followed by Candida ethanolica (13.6%), Kurtzmaniella zeylanoides (9.4%), Geotrichum candidum (8.8%), Galactomyces geotrichum (8.7%), Kluyveromyces lactis (3.5%), and Geotrichum silvicola (2.7%). The volatile profile analysis allowed 24 different compounds to be identified: 7 acids, 7 esters, 4 alcohols, 3 ketones, 2 aromatic hydrocarbons, and 1 aldehyde. The most represented volatile organic compounds (VOCs) were 2-butanone, butanoic acid and hexanoic acid. A positive correlation between Len. kefiri and hexanoic acid and isopentyl isobutyrate was observed (P < 0.05), whereas Y. lipolytica displayed the highest number of positive correlations with 3-methyl-butanal, 2-pentanone and 2-pentanol (P < 0.05). To the authors' knowledge, this is the very first detection of Len. kefiri in a raw ewe's milk cheese coagulated with vegetable rennet., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Among typical Portuguese sausages, the cacholeira blood sausage undoubtedly represents one of the most popular preparations. To the authors' knowledge, a lack of information on both the microbiota and the volatile organic compounds (VOCs) of this blood-containing sausage emerges from the available scientific literature. This study represents the first characterization of physico-chemical, microbiological and volatile traits of Portuguese cacholeira blood sausage. To this end, ready-to-eat cacholeira blood sausages were collected from two production batches manufactured in summer (batch 1) and autumn (batch 2). Viable counts showed active microbial communities mainly composed by lactic acid bacteria, coagulase negative cocci, enterococci and eumycetes. The metataxonomic approach showed a simple bacterial composition, which was dominated by Lactobacillus sakei in both the analyzed batches (1 and 2) considered. Carnobacterium, Enterococcus, Kluyvera, Lactococcus and Serratia were found as minor genera. The mycobiota varied according to the production season. Batch 1 was dominated by Starmerella apicola, Debaryomyces hansenii and Candida tropicalis, whereas batch 2 was dominated by D. hansenii. Moreover, Aspergillus spp., Kurtzmaniella zeylanoides, Saccharomyces cerevisiae, Kurtzmaniella santamariae, Brettanomyces bruxellensis and Pichia kluyveri were detected in both the batches as minority species. Seventy-two volatile compounds were identified, including esters, phenols, terpenoids, acids, alcohols, ketones, aldehydes, lactones, furans, sulphur and nitrogen compounds. Significant differences were seen in the amount of some compounds, as a feasible consequence of differences in the raw materials, artisan production and seasonality., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)