Spanish Cheese Research Articles

Magnetic Resonance Imaging (MRI) of Spanish Sheep Cheese: A Study on the Relationships between Ripening Times, Geographical Origins, Textural Parameters, and MRI Parameters.

The evolution of structural changes and the textural features during the ripening process of four varieties of Spanish sheep cheese were studied using Magnetic Resonance Imaging (MRI). Specifically, longitudinal (T1) and transverse (T2) relaxation times and apparent diffusion coefficient maps were analyzed. Also, proton density was used to improve the description of the structure of the cheeses. The MRI results displayed important information about cheese matrix structure, associated with different manufacturing processes (industrial vs. traditional), ripening times (RTs, from 2 to 180 days), and geographical origins. A significant interaction between RT and cheese variety related to the variations in physicochemical and textural parameters was found. Linear regression models were developed per the abundant literature. Logarithmic regression models showed the highest R2 when monitoring the dependency on T1 and T2 parameters of water content, water activity, RT, and some texture parameters. Therefore, these results support that MRI is a useful technology to monitor the ripening process, predict textural behavior and physicochemical variables, and characterize the structure of different varieties of sheep cheese.

Comparative analysis of the microbiome composition of artisanal cheeses produced in the Mediterranean area

In the PRIMA project ArtiSaneFood, the microbiological parameters of several artisanal cheeses produced in the Mediterranean area have been quantified. In this pilot study, we selected four of these artisanal cheese products from Italy, Portugal, Spain, and Morocco to investigate and compare their microbiomes in terms of taxonomy composition, presence of reads of foodborne pathogens, as well as virulence and antimicrobial resistance genes. Lactococcus, Streptococcus and Lactobacillus were the most represented genera in the Portuguese and Spanish cheeses, Streptococcus in the Italian cheese, and Enterococcus, Klebsiella, Escherichia, and Citrobacter in the Moroccan products. The correlation analysis indicated a negative association between the abundance of some lactic acid bacteria (i.e., Lactococcus, Lactobacillus, Streptococcus, and Leuconostoc) and foodborne pathogenic genera, like Escherichia and Salmonella. The analysis of pathogen abundance, virulence factors, and antimicrobial resistance genes showed a strong clusterization based on the cheese type, confirming that the presence of potential human health risk determinants was higher in the artisanal products derived from unpasteurized milk that underwent spontaneous fermentation.

Advantages and disadvantages of autochthonous enterococci strains for their potential use in cheese ripening: a preliminary study

SummaryThe enzymatic activity exhibited by native enterococci could, in turn, result in a reduction in the cheese ripening time. However, they may also carry virulence factors or resistance to antibiotics that prevent their use in food. A total of 69 enterococci isolates were obtained from a Spanish traditional raw goat's milk cheese and identified by Maldi‐Tof/MS. Different enzymatic activities of technological interest were evaluated and the safety of those with greater technological suitability was studied. The strains were identified as E. malodoratus, E. italicus, E. gilvus, E. faecium, E. durans, E. casseliflavus and E. faecalis. Some strains had very high acidifying and proteolytic activities. Most strains had medium or low amino and carboxypeptidase activities and high dipeptidase activities, while esterase activities were medium. A total of 10 strains were selected based on their enzymatic activities. Of these, five showed β‐haemolysin activity, eight showed gelatinase activity and eight produced tyramine. These activities represent virulence factors that could potentially affect food safety. Finally, all selected strains were susceptible to the antibiotics tested.

Short communication: Comparing the microbiota diversity from the core, middle part and rind of six Spanish commercial goat cheeses

The cheese core has a lower oxygen saturation and salinity and a higher acidity than the rind, but there is controversy about the incidence of such factors on the magnitude of microbial diversity. The goal of the current work was to investigate the existence of differences in α-diversity between the core, middle part, and rind of six Spanish commercial cheeses through a sequencing approach. To this end, we have collected rind, middle part, and core samples from fresh (H and M), soft semi-ripened (C and P), hard semi-ripened (B) and semi-hard aged (G) goat cheeses. After purifying deoxyribonucleic acid from these 18 samples, the V3-V4 ultravariable region of the 16S rRNA gene was sequenced. The analysis of microbial composition revealed that lactic acid bacteria from the genera Lactococcus, Lactobacillus, Streptococcus, and Leuconostoc are predominant in all six goat cheeses. Furthermore, we identified several psychrophilic taxa often associated with the post-pasteurization contamination of refrigerated milk. Comparison of three α-diversity estimators (Chao1, Shannon and Faith's phylogenetic diversity indices) of microbiota in the core, middle part, and rind of all six goat cheeses did not reveal substantial differences, being only significant (at the nominal level) the comparison of rind vs middle part for the Shannon index (P-value = 0.031). Moreover, the construction of a dendrogram based on Aitchison distances revealed that cheese samples cluster according to their manufacturing characteristics, with a clear distinction between fresh vs semi-ripened or aged cheeses. We conclude that the magnitude of microbial α-diversity in the cheese core is similar to that in the rind despite their different physicochemical attributes. This result could be because physicochemical differences between cheese compartments are often attenuated during cheese ripening.

Sensory Attributes and Instrumental Chemical Parameters of Commercial Spanish Cured Ewes' Milk Cheeses: Insights into Cheese Quality Figures.

The external appearance of some of the Protected Designation of Origin (PDO) cured cheeses is similar to other cheese samples made in Spain: 1 kg and 2.5-3 kg formats, cylindrical, and with or without a pleita mark on the surface. In this work, commercial cured ewe's milk cheese samples with a similar external appearance were analyzed, including five PDO and five non-PDO samples. The parameters analyzed were color, texture, pH, humidity, water activity, and the volatile profile. Additionally, a descriptive and consumer-sensory analysis of the cheese samples was carried out. Statistical analysis of the results showed that luminosity, color coordinates a* and b*, percentage of deformation, humidity, water activity, and acid contents were significantly higher in non-PDO cheese samples. The breaking force, maximum force, and the content of esters were significantly higher in those cheese samples with PDO. In addition, PDO cheese samples showed higher scores for all attributes evaluated by consumers, except for color. These results suggest that PDO cheeses are placed on the market with a higher degree of ripening than non-PDO ones and that consequently they are more positively valued by consumers.

Prodigiosin-Producing Serratia marcescens as the Causal Agent of a Red Colour Defect in a Blue Cheese.

Technological defects in the organoleptic characteristics of cheese (odour, colour, texture, and flavour) reduce quality and consumer acceptance. A red colour defect in Cabrales cheese (a traditional, blue-veined, Spanish cheese made from raw milk) occurs infrequently but can have a notable economic impact on family-owned, artisanal cheesemaking businesses. This work reports the culture-based determination of Serratia marcescens as the microbe involved in the appearance of red spots on the surface and nearby inner areas of such cheese. Sequencing and analysis of the genome of one S. marcescens isolate, RO1, revealed a cluster of 16 genes involved in the production of prodigiosin, a tripyrrole red pigment. HPLC analysis confirmed the presence of prodigiosin in methanol extracts of S. marcescens RO1 cultures. The same was also observed in extracts from red areas of affected cheeses. The strain showed low survival rates under acidic conditions but was not affected by concentrations of up to 5% NaCl (the usual value for blue cheese). The optimal conditions for prodigiosin production by S. marscescens RO1 on agar plates were 32 °C and aerobic conditions. Prodigiosin has been reported to possess antimicrobial activity, which agrees with the here-observed inhibitory effect of RO1 supernatants on different bacteria, the inhibition of Enterobacteriaceae, and the delayed development of Penicillium roqueforti during cheesemaking. The association between S. marcescens and the red colour defect was strengthened by recreating the fault in experimental cheeses inoculated with RO1. The data gathered in this study point towards the starting milk as the origin of this bacterium in cheese. These findings should help in the development of strategies that minimize the incidence of pigmenting S. marcescens in milk, the red defect the bacterium causes in cheese, and its associated economic losses.

Large amplitude oscillatory shear stress (LAOStress) analysis for an acid-curd Spanish cheese: Afuega'l Pitu atroncau blancu and roxu (PDO)

Large amplitude oscillatory shear stress (LAOStress) was used to study the nonlinear rheological properties of Afuega'l Pitu (PDO) cheese atroncau blancu and roxu, a Spanish acid-curd cheese made from cow's milk, from nine manufacturers. Composition, physicochemical and lipolytic parameters were determined. The moisture content varied between (23.60 ± 0.75)% and (45.84 ± 0.01)% and the percentage of salt-in-moisture was <(8.17 ± 0.86)%. Stress amplitude sweeps were conducted at 20 °C, 50 °C and 75 °C at frequencies of 0.628 rad/s, 6.28 rad/s and 31.4 rad/s, for each temperature. Experiments were analyzed using the MITlaos software modified to extract LAOStress quantitative material functions. The large number of experiments and the high dimensionality of the LAOS signals were carefully analyzed and reduced to understand the material response and extract the relevant properties. All cheeses showed a distinct signature that motivates us to introduce the concept of “pseudo-linear LAOS” where nonlinear rheology is evident in only some measures while others minimally change from the linear regime. We geometrically interpret these different LAOS measures as rotation and distortion of Lissajous curves; all cheeses showed pseudo-linear LAOS regimes where rotation is strong and distortion is minimal, i.e. first-harmonics change dramatically in a regime where higher-harmonics are small enough that Lissajous curves are nearly elliptical. The pseudo-linear behavior enabled low-dimensional data reduction and three metrics were discovered to be the most relevant: linear elastic modulus (G′), critical stress (σcrit) and strain (γcrit) amplitudes. All Afuega'l Pitu cheese samples softened as stress amplitude increased while maintaining a solid-like behavior (tanδ^1∼ 0.3), regardless of frequency and temperature. Moreover, the difference between blancu and roxu was statistically insignificant compared to difference between manufacturers. Linear G′ and nonlinear σcrit decreased with increasing temperature, while the nonlinear γcrit maintained a constant value at 50 °C and 75 °C. Although G′, σcrit and γcrit metrics establish comparison between cheeses, more metrics are still required to fully connect rheology to cheese texture and sensory profiles.

Effect of Vacuum Packaging on the Biochemical, Viscoelastic, and Sensory Properties of a Spanish Cheese during Chilled Storage

The unique qualities of Spanish cheeses, such as the San Simón da Costa (SSC) cheese, are protected by the Protected Designation of Origin (PDO) status. The technological importance of chilled storage at 4 °C of vacuum-packaged (V) and natural (N) (unpackaged) cheeses was examined. For this purpose, the physico-chemical, biochemical, mechanical (puncture tests), viscoelastic (oscillatory and transient tests) and sensory properties of V and N cheeses were compared and analysed. During chilled storage, the caseins in V cheeses did not undergo proteolytic reactions. Low temperature maintained a low intensity of proteolytic phenomena for up to 6 months. Lipolysis was more intense in the N than in the V samples. The moisture content decreased in the N cheeses during chilled storage, and thus, the casein matrix concentration and ionic strength increased, resulting in an increase in the gel strength (S) parameter and complex modulus (G*), and the conformational stability-high stress amplitude (σmax). The low and similar values of the n' and n'' exponents (mechanical spectra) and the n parameter (transient tests) indicated the high degree of the temporal stability of the cheese network in both the N and V samples, irrespective of storage time. Likewise, the similar values of the phase angle (δ) for the N and V cheeses during storage indicate energy-stable bonds in the SSC cheese matrix. The attributes of the oral tactile phase (firmness, friability, gumminess, and microstructure perception), mechanical parameters and viscoelastic moduli enabled the discrimination of the packaged and unpackaged cheeses. Cheeses chilled and stored without packaging were awarded the highest scores for sensory attributes (preference) by trained panellists.

Changes in the Concentration and Profile of Free Fatty Acids during the Ripening of a Spanish Blue-Veined Cheese Made from Raw and Pasteurized Cow and Goat Milk

Blue-veined cheeses in general undergo a pronounced lipolysis. The aim of this work was to determine the evolution of free fatty acids (FFA) in Valdeón cheese during ripening, comparing cheeses made from raw and pasteurized milk. The effect of season on the evolution of FFA in pasteurized milk cheeses was also studied. Cheeses made with raw milk showed the highest concentrations of FFA, reaching values of 23,081.9 mg 100 g−1 dry matter at the end of ripening, compared to the values of cheeses made with pasteurized milk (7327.1 mg 100 g−1 dry matter), in both cases with a predominance of oleic and palmitic acids. However, pasteurization did not affect the FFA profile of the cheeses. Regarding the cheeses made with pasteurized milk in different seasons, the highest FFA concentration was reached in cheeses made in summer after 30 days of ripening. The season also influenced the FFA profile and thus the concentration of short-, medium- and long-chain fatty acids in relation to total FFA. There were no significant differences in sensory analysis between cheeses made from raw and pasteurized milk.

Lactose and Galactose Content in Spanish Cheeses: Usefulness in the Dietary Treatment of Patients with Galactosaemia.

In galactosaemia, a strict galactose-free diet is necessary to prevent or resolve acute symptoms in infants. However, because the body produces up to 10 times more galactose than is found in a galactose-restricted diet, excessively restrictive diets should be avoided in children and adults to prevent nutritional deficiencies. Since cheese is a nutritional source of the calcium necessary for bone health, the latest international guidelines on the management of classical galactosaemia (2017) allow the consumption of cured cheeses with less than 25 mg of galactose/100 g and recommend that each country verifies the adequacy of the cheeses, since most mature cheeses do not always have a lower galactose content. In total, 32 cheese samples were purchased (19 Spanish and 13 international cheeses), and their lactose and galactose contents were analysed using ion chromatography with pulsed amperometric detection (IC-PAD). Five Spanish cheeses contained less than 25 mg of galactose/100 g: García Baquero semi-cured cheese; Hacendado, Gran Reserva and Mahón cured cheeses; and García Baquero Reserva 12-month cured cheese. In addition, eight international cheeses were confirmed as suitable: Comté, Gouda, Gruyere, Maasdam, Parmigiano, Edam, Emmental, and some samples of Cheddar. In addition to the well-known low-galactose Swiss and Dutch cheeses, according to the current results, five Spanish cheeses can be safely consumed. The greater availability of types of cheese favours better bone health in patients with galactosaemia.

Characterization of autochthonal Hafnia spp. strains isolated from Spanish soft raw ewe's milk PDO cheeses to be used as adjunct culture

The present work was performed to study the enterobacteria involved in the ripening of the artisanal raw ewe's milk PDO cheeses ‘Torta del Casar’ and ‘Queso de la Serena’ produced in Extremadura (Spain). These isolates were strain-typed, safety tested and characterized for some important technological properties. A total of 485 enterobacterial isolates were clustered by RAPD-PCR and subsequently identified by partial sequencing of the 16S rRNA gene. Among the 17 different species identified, Hafnia paralvei was the predominant species; H. alvei and Lelliottia amnigena were present to a lesser extent. Therefore, 55 Hafnia spp. strains, selected according to their genetic profile and dairy origin, were tested for the safe application. Overall, they were able to produce the biogenic amines putrescine and cadaverine under favourable conditions, presented α-haemolytic activity and did not produce cytolytic toxin active against HeLa cells or contain virulence genes. In addition, antibiotic susceptibility profiles showed that 17 Hafnia spp. strains were less resistant to the 33 antibiotics tested; subsequently, they were further technologically characterized. Although they showed differences, in general, they were well adapted to the stress conditions of cheese ripening. Among them, two strains, H. alvei 544 and 1142, are highlighted mainly due to their proteolytic activity at refrigeration temperatures and their low or null gas production. Although further studies are necessary before industrial application, these two strains are proposed for potential use as adjunct cultures to favour the homogeneity of these PDO cheeses, preserving their unique sensory characteristics.

Isolation and phenotypic and genomic characterization of Tetragenococcus spp. from two Spanish traditional blue-veined cheeses made of raw milk

High throughput sequencing has recently revealed the presence of Tetragenococcus-related DNA sequences in dairy environments such as brine and cheeses. In the present work, a selective medium was developed to isolate Tetragenococcus spp. from two ripened, traditional, Spanish, blue-veined cheese varieties made from raw milk. The strains recovered belonged to either Tetragenococcus koreensis or Tetragenococcus halophilus species. Twenty of these isolates (15 of T. koreensis and 5 of T. halophilus) were then subjected to a battery of phenotypic and genetic tests, and six strains (4 T. koreensis and 2 T. halophilus) to genome sequencing. Wide genetic and phenotypic diversity was noted. All strains grew poorly in milk, producing small quantities of lactic and acetic acids. Most strains used lactose as a carbon source and ferment milk citrate. In agreement, genome analysis detected in the genome of the six strains analyzed gene clusters harboring several lactose/galactose-related genes and genes encoding citrate metabolic enzymes (permease, citrate lyase, and oxaloacetate decarboxylase). Most of the tested strains were resistant to erythromycin and clindamycin, and a few to other antimicrobial agents, but neither known mutations nor acquired genes conferring resistance to antibiotics were identified in their genomes. Neither were genes coding for pathogenicity or virulence factors detected. Decarboxylase-encoding genes involved in biogenic amine production were not identified, in keeping with the strains' negative biogenic amine-producer phenotype. Genome comparison revealed vast arrays of genes (similar in number to those described in other lactic acid bacteria) coding for components of proteolytic and lipolytic systems. Tetragenococcus strains showing desirable traits plus the absence of detrimental features might be exploitable in the form of secondary, adjunct or ripening cultures to ensure the typical bouquet of traditional blue-veined cheeses is obtained, or to diversify the final flavor in other varieties.

Characteristics and proteolysis of a Spanish blue cheese made with raw or pasteurised milk

Valdeón cheese is a Spanish Protected Geographical Indication of blue‐veined cheese produced on an industrial scale from raw or pasteurised cow and/or goat milk. The aim of this work was to evaluate the impact of pasteurisation on the microbiological, physicochemical and sensory characteristics of cheese. Cheeses made with raw milk showed higher counts of lactobacilli and enterococci, as well as lower values of pH and D‐lactic acid and salt/moisture ratio. Cheeses made with pasteurised milk showed greater extent of proteolysis, higher concentration of free amino acids and lower concentration of biogenic amines. Pasteurisation produced more elastic and harder cheeses, but with a lower sensory profile.

Control of Listeria monocytogenes growth and virulence in a traditional soft cheese model system based on lactic acid bacteria and a whey protein hydrolysate with antimicrobial activity

“Torta del Casar” is a Spanish soft-ripened cheese made with sheep's raw milk and subjected to a short ripening process, which favors the growth of pathogenic microorganisms including Listeria monocytogenes. The development of strategies to control pathogens and minimize health risks associated with the presence of L. monocytogenes in these products is of great interest. In this regard, the anti-Listeria activity of a whey protein hydrolysate (ProH) alone or combined with six lactic acid bacteria strains isolated from cheese was evaluated in this study as a biocontrol strategy using a “Torta del Casar” cheese-based medium. The most active combinations of lactic acid bacteria assayed induced a reduction higher than two logarithmic units in the growth of L. monocytogenes (serotype 4b) compared to their respective control when they were co-inoculated in “Torta del Casar” cheese-based medium at 7 °C for 7 days. In addition, the observed downregulation of some key virulence genes of L. monocytogenes suggests that the strain Lactiplantibacillus plantarum B2 alone and combined with the strain Lactiplantibacillus spp. B4 are good candidates to be used as biocontrol agents against L. monocytogenes growth in traditional soft cheeses based on raw milk during their storage at refrigeration temperatures.

Changes in thermoviscoelastic and biochemical properties of Atroncau blancu and roxu Afuega'l Pitu cheese (PDO) during ripening

Thermoviscoelastic and biochemical properties of Afuega'l Pitu cheese blancu (B) and roxu (R), a Spanish acid-curd cheese made from cow’s milk, were determined at 3, 15, 30 and 60 days of ripening. The contents of fat (>45 g/100 g total solids, TS) and NaCl (<3 g/100 g TS) were similar between varieties and did not change with ripening. Total solids, pH, nitrogen fractions and fat acidity increased with ripening time in B and R samples. Lactose content was higher in B than in R cheeses at 3 d, was degraded gradually throughout ripening but did not disappear totally in both cheeses at 60 d. Viscoelastic parameters were determined in the linear viscoelastic region (LVER) by stress sweeps (6.3 rad/s, 20 °C): the stress amplitude (σmax) and complex modulus (G*) increased, while strain amplitude (γmax) decreased up to 60 d. Mechanical spectra were also determined, in the LVER, at 20 °C, 50 °C and 75 °C, showing that all samples behaved as “true gels” (elastic modulus, G' > viscous modulus, G''), irrespective of temperature. The best gel properties (low values of loss factor, tanδ = G''/G', and minor frequency dependence, i.e., low n', n'' exponents), were observed at 20 °C for B30 and R30. The viscoelastic parameters (G0' and G0'') increased with ripening time, irrespective of the temperature, and for a fixed ripening time they were lower at higher temperature. At 75 °C, all samples exhibited notable frequency dependence (n'' ≫ n'), resulting in shear-induced gelation at low frequencies (high oscillation times) which was consistent with data in thermal profiles from 70 °C to 90 °C.

Behaviour of Non-O157 STEC and Atypical EPEC during the Manufacturing and Ripening of Raw Milk Cheese.

This study was carried out to assess the survival of Shiga toxin-producing E. coli (STEC) and atypical enteropathogenic Escherichia coli (aEPEC) during the traditional manufacturing and ripening of Spanish hard cheese from raw cow’s milk. Milk samples were spiked with up to 3.1–3.5 log cfu/mL of one strain of STEC (O140:H32 serotype) and one of aEPEC (serotype O25:H2). The first steps of cheesemaking allow for a STEC and aEPEC increase of more than 1 log cfu/mL (up to 4.74 log cfu/g and 4.55 log cfu/g, respectively). After cheese pressing, a steady reduction of both populations was observed, with the STEC strain being more sensitive. The studied pathogenic E. coli populations decreased by 1.32 log cfu/g in STEC and 0.59 log cfu/g in aEPEC in cheese ripened during a minimum period of 60 d. Therefore, a moderate contamination by these diarrhoeagenic E. coli pathotypes, in particular, with aEPEC, on cheese manufactured from raw milk may not be totally controlled through the cheesemaking process and during a maturation of 90 d. These findings remark the importance of improvement in bacteriological quality of raw milk and cross-contamination prevention with diarrhoeagenic E. coli in the dairy industry.

Human Bifidobacterium strains as adjunct cultures in Spanish sheep milk cheese

Three bifidobacteria strains of human origin (Bifidobacterium pseudolongum INIA P2, Bifidobacterium breve INIA P734, and Bifidobacterium longum INIA P678) were used as potential probiotic adjunct cultures for the manufacture of pasteurized sheep milk cheese. Bifidobacteria were inoculated at 5 to 6 log cfu/mL in milk vats. Microbiological, physicochemical, rheological, color, and sensory characteristics were determined at 7, 28, and 60 d of ripening. Counts of B. pseudolongum INIA P2 remained above 6 log cfu/g during 60 d of ripening as well as after further simulated gastrointestinal digestion of cheeses. Bifidobacterium breve INIA P734 counts remained stable during 28 d and decreased by less than 1 log unit after simulated digestion. Bifidobacterium longum INIA P678 counts dropped sharply during cheese manufacture and ripening and were below detection level after simulated digestion. Addition of bifidobacteria strains did not affect starter viability, cheese pH, dry matter, water activity, or salt content but significantly increased overall proteolysis and the concentration of some free amino acids. Cheeses with bifidobacteria exhibited no significant differences in most sensory characteristics with respect to control cheese. According to our results, B. breve INIA P734 and B. pseudolongum INIA P2 are promising candidates as probiotic adjunct cultures in fresh and semi-hard sheep milk cheese.

Identification of Species and Subspecies of Lactic Acid Bacteria Present in Spanish Cheeses Type "Torta" by MALDI-TOF MS and pheS gene Analyses.

Several artisanal cheeses are elaborated in European countries, being commonly curdled with rennets of animal origin. However, in some Spanish regions some cheeses of type “Torta” are elaborated using Cynara cardunculus L. rennets. Two of these cheeses, “Torta del Casar” and “Torta de Trujillo”, are elaborated in Cáceres province with ewe’s raw milk and matured over at least 60 days without starters. In this work, we identified the lactic acid bacteria present in these cheeses using MALDI-TOF MS and pheS gene analyses, which showed they belong to the species Lactobacillus curvatus, Lactobacillus diolivorans, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactococcus lactis and Leuconostoc mesenteroides. The pheS gene analysis also allowed the identification of the subspecies La. plantarum subsp. plantarum, La. paracasei subsp. paracasei and Le. mesenteroides subsp. jonggajibkimchii. Low similarity values were found in this gene for some currently accepted subspecies of Lc. lactis and for the two subspecies of La. plantarum, and values near to 100% for the subspecies of Le. mesenteroides and La. paracasei. These results, which were confirmed by the calculated ANIb and dDDH values of their whole genomes, showed the need to revise the taxonomic status of these species and their subspecies.

Polyphasic identification of Penicillium spp. isolated from Spanish semi-hard ripened cheeses

Fifteen samples of semi-hard ripened cheeses, both spoiled (10) and unspoiled (5), and obtained from cheese factories located in Northwest of Spain, were analysed by a dilution plating technique and direct sampling. A total of 32 isolates were identified at species level by a polyphasic approach (phenotypic characterization, partial extrolite analysis and molecular identification). Most isolates (65.6%) belonged to the species P. commune; other species found were P. solitum, P. chrysogenum, P. nordicum, P. expansum and P. cvjetkovicii. All of the P. commune isolates were able to produce cyclopiazonic acid, while the P. nordicum and the P. expansum isolates were producers of ochratoxin A and patulin respectively. Despite this, the role of P. commune as beneficial fungi in cheese ripening should be investigated. Molecular identification based on BenA sequence analysis was able to identify the majority of isolates. The three mycotoxins investigated can be considered key for identification. The polyphasic approach seems to be a very valuable tool for identification of isolates of this complex genus.

Convenções de qualidade em queijos artesanais no Brasil, Espanha e Itália

Muitas vezes os alimentos artesanais são vistos pelos consumidores como sendo mais naturais e saudáveis e são valorizados devido a sua tradição, origem e sustentabilidade dos processos envolvidos. Os queijos artesanais representam bem essa categoria de alimentos, pois sua caracterização envolve o tipo de matéria-prima, o método de processamento, as características edafoclimáticas de onde são produzidos, as receitas utilizadas, a pequena escala de produção, as relações estabelecidas entre o produtor e o consumidor. No entanto, essas características estão balizadas por normas sanitárias que nem sempre conseguem ser atendidas pelos produtores, que acabam por realizar a venda informalmente. O objetivo deste artigo é demonstrar as diferenças existentes entre as concepções, práticas e estratégias de qualidade de queijos artesanais e o impacto da legislação sanitária sobre eles a partir de estudos de caso específicos de queijarias do Brasil, Espanha e Itália.