Brazilian Cheese Research Articles

Probiotic Potential and Application of Indigenous Non-Starter Lactic Acid Bacteria in Ripened Short-Aged Cheese.

There are massive sources of lactic acid bacteria (LAB) in traditional dairy products. Some of these indigenous strains could be novel probiotics with applications in human health and supply the growing needs of the probiotic industry. In this work, were analyzed the probiotic and technological properties of three Lactobacilli strains isolated from traditional Brazilian cheeses. In vitro tests showed that the three strains are safe and have probiotic features. They presented antimicrobial activity against pathogenic bacteria, auto-aggregation values around 60%, high biofilm formation properties, and a survivor of more than 65% to simulated acid conditions and more than 100% to bile salts. The three strains were used as adjunct cultures separately in a pilot-scale production of Prato cheese. After 45days of ripening, the lactobacilli counts in the cheeses were close to 8 Log CFU/g, and was observed a reduction in the lactococci counts (around -3 Log CFU/g) in a strain-dependent manner. Cheese primary and secondary proteolysis were unaffected by the probiotic candidates during the ripening, and the strains showed no lipolytic effect, as no changes in the fatty acid profile of cheeses were observed. Thus, our findings suggest that the three strains evaluated have probiotic properties and have potential as adjunct non-starter lactic acid bacteria (NSLAB) to improve the quality and functionality of short-aged cheeses.

Thermosonication as an effective substitution for fusion in Brazilian cheese spread (Requeijão Cremoso) manufacturing: The effect of ultrasonic power on technological properties

In this initial study, the impact of thermosonication as an alternative to the traditional fusion in Brazilian cheese spread (Requeijão Cremoso) manufacture was investigated. The effect of ultrasound (US) power was evaluated considering various aspects such as gross composition, microstructure, texture, rheology, color, fatty acid composition, and volatile compounds. A 13 mm US probe operating at 20 kHz was used. The experiment involved different US power levels (200, 400, and 600 W) at 85 °C for 1 min, and results were compared to the conventional process in the same conditions (85 °C for 1 min, control treatment). The texture became softer as ultrasound power increased from 200 to 600 W, which was attributed to structural changes within the protein and lipid matrix. The color of the cheese spread also underwent noticeable changes for all US treatments, and treatment at 600 W resulted in increased lightness but reduced color intensity. Moreover, the fatty acid composition of the cheese spread showed variations with different US power, with samples treated at 600 W showing lower concentrations of saturated and unsaturated fatty acids, as well as lower atherogenicity and thrombogenicity indexes, indicating a potentially healthier product. Volatile compounds were also influenced by US, with less compounds being identified at higher powers, especially at 600 W. This could indicate possible degradation, which should be evaluated in further studies regarding US treatment effects on consumer perception. Hence, this initial work demonstrated that thermosonication might be interesting in the manufacture of Brazilian cheese spread, since it can be used to manipulate the texture, color and aroma of the product in order to improve its quality parameters.

Comparison of hyperspectral imaging and spectrometers for prediction of cheeses composition

Artisanal cheeses are part of the heritage and identity of different countries or regions. In this work, we investigated the spectral variability of a wide range of traditional Brazilian cheeses and compared the performance of different spectrometers to discriminate cheese types and predict compositional parameters. Spectra in the visible (vis) and near infrared (NIR) region were collected, using imaging (vis/NIR-HSI and NIR-HSI) and conventional (NIRS) spectrometers, and it was determined the chemical composition of seven types of cheeses produced in Brazil. Principal component analysis (PCA) showed that spectral variability in the vis/NIR spectrum is related to differences in color (yellowness index) and fat content, while in NIR there is a greater influence of productive steps and fat content. Partial least squares discriminant analysis (PLSDA) models based on spectral information showed greater accuracy than the model based on chemical composition to discriminate types of traditional Brazilian cheeses. Partial least squares (PLS) regression models based on vis/NIR-HSI, NIRS, NIR-HSI data and HSI spectroscopic data fusion (vis/NIR + NIR) demonstrated excellent performance to predict moisture content (RPD > 2.5), good ability to predict fat content (2.0 < RPD < 2.5) and can be used to discriminate between high and low protein values (∼1.5 < RPD < 2.0). The results obtained for imaging and conventional equipment are comparable and sufficiently accurate, so that both can be adapted to predict the chemical composition of the Brazilian traditional cheeses used in this study according to the needs of the industry.

Gelatin-based films incorporated with buriti oil (Mauritia flexuosa L.) as active packaging for artisanal cheese conservation

Active and biodegradable materials have attracted interest in the food packaging area for better product safety and quality. In this study, a gelatin-based film was developed incorporating buriti oil extracted by pressurized propane to preserve the coalho artisanal Brazilian cheese. The GC/qMS analysis of buriti oil revealed unsaturated fatty acids, polyphenols, and carotenoids high content. No significant differences in film thickness, elongation at break, tensile strength, and Young's module were observed between oil-based and oil-free gelatin films. However, oil incorporation into the film decreased the water vapor permeability. Oil-incorporated gelatin-based films displayed antioxidant, bacteriostatic properties and pH responsivity during cheese storage compared to control groups. This trial revealed a significant growth decrease of Salmonella spp., P. aeruginosa, and E. coli, over 14-day storage at 5 °C, indicating the minimized microbial cheese spoilage during storage and the promising application of films for food-active packaging.

Evaluation of traditional semi‐hard Brazilian cheeses using chemometric tools to identify possible denomination fraud

This study aimed to evaluate and classify some of the main traditional Brazilian semi‐hard cheeses, using multivariate tools to determine possible denomination fraud. Fifty‐six samples of Minas Artisanal (MA), Colonial (AC), Minas Padrão (MP) and Meia Cura (MC) cheeses were analysed. All the evaluated parameters indicated high variability. PCA and HCA confirmed two groupings (AC and MP; and MA and MC) with high similarity. The DD‐SIMCA method classified the cheeses with 83% accuracy. AC (48%) and MA (20%) cheeses were the most well classified in their respective denomination. These results were efficient in classifying groups and demonstrating the lack of authenticity regarding the denominations of these cheeses.

Technological Characterization of Lactic Acid Bacteria Strains for Potential Use in Cheese Manufacture.

A total of 26 lactic acid bacteria isolates from both Italian and Brazilian cheeses were tested for their use in cheesemaking. Isolates were screened for salt tolerance, exopolysaccharide and diacetyl production, lipolytic, acidifying, and proteolytic activities. In addition, the aminopeptidase (Pep N and Pep X) activities, were evaluated. Most of the strains demonstrated salt tolerance to 6% of NaCl, while only two L. delbruekii (P14, P38), one L. rhamnosus (P50) and one L. plantarum (Q3C4) were able to grow in the presence of 10% (w/v) of NaCl. Except for 2 L. plantarum (Q1C6 and Q3C4), all strains showed low or medium acidifying activity and good proteolytic features. Furthermore, lipolytic activity was revealed in none of the strains, while the production of EPS and diacetyl was widespread and variable among the tested strains. Finally, regarding aminopeptidase activities, 1 L. delbrueckii (P10), 1 L. rhamnosus (P50), and 1 L. lactis (Q5C6) were considered as the better performing, showing high values of both Pep N and Pep X. Based on data presented here, the aforementioned strains could be suggested as promising adjunct cultures in cheesemaking.

THE MEDIATING EFFECT OF PERSONAL VALUES ON THE INFLUENCE OF HUMAN CAPITAL ON THE PRODUCTION OF ARTISANAL CHEESES

This study proposed the identification of personal values and their role as mediation between human capital and the expression of quality. The relationship between these factors in artisanal food production in small companies is still little explored, so this work has the objective of bringing advances in this theme. For the analysis of the results, a quantitative method was used, structural model equations, through the partial least squares regression method (PLS). The results found were satisfactory, demonstrating that investments in human capital must be the object of public policies, as their influence overlaps the influence of the personal values of each entrepreneur for the expression of quality conventions. The assumptions about the effect of mediation of values in the relationship between human capital and quality worlds are positive and significant, demonstrating that the values had a complementary partial mediation effect between human capital and quality, but with less importance than the direct impact of human capital in quality. It was possible to verify the importance of personal experience, combined with constant training for the development of human capital in the company. In small companies, as is the case with the production of artisanal Brazilian cheeses, in which production and handling are under the control of the family, the contribution of human capital to quality is even more evident. At the same time, personal values have a strong influence on small companies, where the characteristics, behavior, attitudes and beliefs of the entrepreneur are decisive for the results of the enterprise.

Revealing the microbial heritage of traditional Brazilian cheeses through metagenomics

Brazilian artisanal cheeses date from the first Portuguese settlers and evolved via local factors, resulting in unique products that are now part of the patrimony and identity of different Brazilian regions. In this study, we combined several culture-independent approaches, including 16S/ITS metagenetics, assembly- and deep profiling-metagenomics to characterize the originality of the microbiota of five varieties of Brazilian artisanal cheeses from the South and Southeast regions of Brazil. Their core microbiota contained mainly lactic acid bacteria (LAB), of which Lactococcus lactis subsp. lactis was the most frequent, followed by Streptococcus thermophilus in the South region. Moreover, several samples from the Southeast region contained, as dominant LAB, two other food Streptococci belonging to a new species of the salivarius group and S. infantarius. Rinds of samples from the Southeast region were dominated by the halotolerant bacterium Corynebacterium variabile, and the yeasts Diutina catenulata, followed by Debaryomyces hansenii and Kodamaea ohmeri. Rinds from the South region contained mainly LAB due to their short ripening time, and the predominant yeast was D. hansenii. Phylogenomic analysis based on L. lactis metagenome-assembled genomes (MAGs) showed that most Brazilian strains are closely related and form a different clade from those whose genomes are available at this time, indicating that they belong to a specific group. Lastly, functional analysis showed that S. infantarius acquired a ∼ 26 kb DNA fragment from S. thermophilus starter strains that carry the LacSZ system, allowing fast lactose assimilation, an adaptation advantage for growth in milk. Finally, our study identified several areas of concern, such as the presence of somatic cell DNA and high levels of antibiotic resistance genes in several cheese microbiota, suggesting that milk from diseased animals may still be used occasionally. Overall, the data from this study highlight the potential value of the traditional and artisanal cheese production network in Brazil, and provide a metagenomic-based scheme to help manage this resource safely.

Identification and characterisation of lactobacilli isolated from an artisanal cheese with antifungal and antibacterial activity against cheese spoilage and mycotoxigenic Penicillium spp.

Given the importance of lactic acid bacteria (LAB) in dairy products, this study assessed the antimicrobial properties of lactobacilli against some toxigenic Penicillium spp and pathogenic bacteria commonly found in cheese. Lactobacilli previously isolated from an artisanal Brazilian cheese (n = 32) were identified to species level by amplification and sequencing of the 16S rRNA gene and tested against Penicillium commune, Penicillium expansum, Penicillium nordicum and Penicillium roqueforti through an overlay method. The most resistant mould was P. commune-M35 and the major species that exhibited inhibition against all Penicillium spp. tested was Lactiplantibacillus plantarum. Cell free supernatants of two selected L. plantarum isolates (L119 and L49) were used to determine the antifungal and antibacterial activity using impedance and turbidimetric techniques, respectively. The antimicrobial activity showed inhibition beyond acid. Since L. plantarum-L49 and L119 have the ability to inhibit undesirable microorganisms, they could be candidates as novel protective cheese cultures.

Typical Brazilian cheeses: safety, mineral content and adequacy to the nutritional labeling

This study aimed to evaluate the physicochemical and microbiological characteristics of six typical Brazilian cheeses (bovine milk coalho, goat milk coalho, fresh ricotta, manteiga, fresh Minas, and Padrão Minas). Padrão Minas cheese presented high acidity and the highest contents of protein, fat, and ash, while Ricotta cheese presented the lowest contents of protein, fat, and ash. Bovine milk coalho and Minas fresh cheeses were characterized as source of zinc. Manteiga cheese was characterized by the yellow color (higher b* values). Goat milk coalho cheese was characterized by the white color (higher L* values) and presented the highest concentration of minerals (potassium, iron, magnesium, calcium, manganese, and phosphorus). Part of the typical Brazilian cheeses did not agree with the legislation for moisture (5.9%) and fat in the dry matter (11.8%) classifications, as well as, did not have adequate concentrations of protein (65.4%) and fat (44.9%) stated in the label, being the non-conformity dependent on the cheese type. Furthermore, 30% of the cheeses (30%) disagreed with the microbiological Brazilian legislation, as they presented coliforms and/or coagulase-positive Staphylococcus counts above the limits. It can be concluded that the Brazilian cheeses are nutritive, but the microbial quality and nutritional labels need improvement.

Insights into Blue Discoloration in Brazilian Fresh Soft Cheese: Comparative Genomic and Functional Analyses on Pseudomonas Spp. Strains

Insights into Blue Discoloration in Brazilian Fresh Soft Cheese: Comparative Genomic and Functional Analyses on Pseudomonas Spp. Strains

Presence of Staphylococcus aureus and staphylococcal enterotoxin A Production and Inactivation in Brazilian Cheese Bread

HIGHLIGHTS The study shows the S. aureus counts of the 100 frozen cheese bread samples analyzed. 27 samples (27.0%) had S. aureus counts greater than 5.0 × 103 CFU/g. None of the 62 S. aureus strains isolated carried the genes sea, seb, sec, or sed. Heat treatment at 180 °C for 35 minutes was not sufficient to inactivate SEA.

Bioavailability evaluation of calcium, magnesium and zinc in Brazilian cheese through a combined model of in vitro digestion and Caco-2 cells

Cheese has important nutritional properties as a result of their protein, lipid and essential minerals content. This study analyzed the bioavailability of Ca, Mg and Zn in Brazilian cheeses, using the in vitro INFOGEST digestion method coupled with assays in Caco-2 cells. Furosine content (marker of the early Maillard reaction) was also evaluated. Two self-produced fresh cheeses, cow and goat Minas frescal; and two commercial matured goat cheeses, Blue and Pyramid, were analyzed. Mineral bioaccessibility after in vitro digestion was in the range of 44.4–74.7 %, 54.8–66.1 % and 18.2–38.7 % for Ca, Mg and Zn, respectively. Bioaccessibilty was significantly affected by the mineral and fat content level. The digested fresh cheeses (Minas frescal) showed the highest values of soluble Ca and Mg; and the Pyramid goat cheese presented the greatest Ca and Zn transport efficacy across Caco-2 cell monolayers. Furosine could be a useful indicator to evaluate cheese ripeness and its indirect effect on the mineral availability.

Use of encapsulated lactic acid bacteria as bioprotective cultures in fresh Brazilian cheese.

There is great interest for biopreservation of food products, and encapsulation may be a good strategy to extend the viability of protective cultures. In this study, Lactobacillus paraplantarum FT-259 and Lactococcus lactis QMF 11 were separately encapsulated in casein/pectin (C/P) microparticles, which were tested for antilisterial and anti-staphylococcal activity in fresh Minas cheese (FMC) stored at 8°C. The encapsulation efficiency for both lactic acid bacteria (LAB) was 82.5%, with viability over 6.2 log CFU/g after storage of C/P microparticles for 90days under refrigeration. Interestingly, free Lb. paraplantarum and free Lc. lactis grew significantly in refrigerated FMC, both in the presence and absence of pathogens, but only the first significatively grew when encapsulated. Encapsulation increased the antilisterial activity of Lb. paraplantarum in FMC. Moreover, Lc. lactis significantly inhibited listerial growth in FMC in both its free and encapsulated forms, whereas Staphylococcus aureus counts were only significantly reduced in the presence of free Lc. lactis. In conclusion, these results indicate that C/P microparticles are effective carriers of LAB in FMC, which can contribute for the assurance of the safety of this product.

Assessment of Viral Contamination of Five Brazilian Artisanal Cheese Produced from Raw Milk: a Randomized Survey.

Enteric viruses have been described as important contaminants in fresh and ready-to-eat foods such as sandwiches, deli meat and dairy products. This is a cross-sectional randomized survey to estimate the prevalence of norovirus and human adenovirus (HAdV) from 100 Brazilian artisanal raw milk cheese samples (Minas and Coalho) obtained from different agroindustries in four producing regions in the states of Minas Gerais and one in Piauí, respectively. From October 2017 to April 2018, norovirus genogroups I and II and HAdV were investigated in these cheese samples by RT-qPCR and qPCR, respectively. Viruses were detected in 43 samples, being 26 norovirus GI strains, 14 HAdV, and 3 both viruses. Norovirus GII strains were not detected. Viral concentrations ranged from 6.17 × 104 to 1.44 × 107 genome copies/L-1 and murine norovirus 1 used as internal process control showed 100% success rate of recovery with efficiency of 10%. There was a trend towards a higher positivity rate for both viruses in the rainy season, and HAdV were more commonly found among samples with higher fecal coliform counts. This study is a first step in assessing the risk that this contamination may pose to the consumer of raw products as well as emphasizing the need for good manufacturing practices, quality control systems in the dairy industry and markets. As a randomized survey, we established baseline figures for viruses' prevalence in five types of ready-to-eat raw milk artisanal Brazilian cheese, to allow any monitoring trends, setting control targets and future local risk analyses studies.

Artisanal Brazilian Cheeses-History, Marketing, Technological and Microbiological Aspects.

This review focused on the historical, marketing, technological, and microbiological characteristics of artisanal Brazilian cheese. Brazilian cheese production was introduced and developed from the influence of immigrants considering the combination of climate, races of the animals, quality and specificity of milk, technological cheese-making processes and environmental microbiology, among other factors. It resulted in cheese products with specific physicochemical, microbiological, and sensory quality, which represent the heritage and identities of the different Brazilian regions. The production of artisanal cheese increased in many Brazilian regions, mainly in the southeast, especially due to the traditional production and innovative development of new varieties of cheese. The microbiological quality and safety of raw-milk artisanal cheese continues to be a concern and many studies have been focusing on this matter. Special attention needs to be given to the cheeses produced by raw milk, since numerous reports raised concerns related to their microbiological safety. This fact requires attention and the implementation of strict hygiene practices on the production and commercialization, besides appropriate governmental regulations and control. However, more studies on the relationship between technological processes and microbiological properties, which results in a superior culinary quality and safety of artisanal Brazilian cheeses, are needed.

Chemometric evaluation of the metabolites and volatile profiles of mite-ripened cheeses

Cheese ripening in the presence of mites is a common practice in some European countries. In Brazil, this practice is incipient and it is still unknown whether the mite diversity and mite-mediated processes in the production of Brazilian cheeses are important for product differentiation. Here, we conducted chemometric evaluations of the volatile organic compounds (VOCs) and metabolites in two types of Brazilian cheeses (Minas and Colonial) ripened in the presence and absence of mites, using gas chromatography–mass spectrometry and nuclear magnetic resonance spectroscopy. The presence of mites during cheese ripening influenced the development of fruity VOCs on the surface (rind) of both the cheeses. However, a few influences of mite were observed in the core of cheeses. The mite presence induced the production of metabolites and VOCs that conferred sensory identity to the cheeses.

Artisan minas cheese of Serro: proteolysis during ripening

The Artisan Minas Cheese (AMC) is the oldest and most traditional Brazilian cheese, it is produced in several regions of the state of Minas Gerais, such as the Serro region. The most striking features of the AMC-Serro are the use of raw milk and natural bacteria from the whey, popularly known as pingo, as well as the use of the rind washing process. The aim of the present study was to evaluate the proteolysis of the AMC-Serro from three different producers, during 60 days of maturation, and to relate the proteolysis to the producing farms, the production season and the rind washing during ripening. For this purpose, TRICINE-SDS-PAGE, proteolysis extension and depth indices, moisture, and texture (firmness) were evaluated. It was concluded that the temperature and moisture of the cheeses, that were determined by the location of the ripening room, the production season and the rind washing, were the most important factors. The degree of proteolysis also had an impact on the water loss during ripening, with effect on cheese safety. The results obtained in this study may be used to better understand the transformations during ripening of AMC-Serro and help the small traditional farmers to improve their product's quality and stability.

Enterococcus spp. in Brazilian artisanal cheeses: Occurrence and assessment of phenotypic and safety properties of a large set of strains through the use of high throughput tools combined with multivariate statistics

Traditionally fermented food generally has unique sensorial aspects due to its high microbial diversity, being broadly consumed worldwide. Despite this, some Vancomycin-Resistant-Enterococci (VRE) can occur in these products, which raises questions about their safety. Thus, this study tries to assess information about the occurrence of Enterococcus strains in traditional Brazilian cheese samples collected from the five main regions as well as the characterization of their phenotypic (proteolytic, lipolytic and bacteriogenic potential) and safety (hemolysin activity and growth at commercial VRE selecting media) aspects. A total of 1669 lactic acid bacteria were isolated, of which 605 (36.2%) were identified as Enterococcus by 16sRNA sequencing. Out of these, E. faecium (58.7%) was identified as the dominant species, followed by Enterococcus faecalis (31.8%), Enterococcus durans (4.8%), Enterococcus hermanniensis (1.6%) and Enterococcus gilvus (1.6%). All the data were analyzed by multivariate statistics, and the findings highlighted the presence of Enterococcus as a naturally component of indigenous microbiota of Brazilian artisanal cheeses, mostly in the cheeses of Marajó, Coalho, and Manteiga, possibly as a consequence of their heat resistance and the curd cooking step applied during the process in these regions. The majority of the identified Enterococcus (63.1%) did not produce β-hemolysin and were not resistant to vancomycin. Interestingly some of these strains (21.4%) were able to inhibit Listeria monocytogenes ATCC 35152, mostly isolates from Serrano, Colonial, and Minas artisanal (from Cerrado microregion) cheeses. Also, 10.2 and 11.8% of them have shown moderate to high proteolytic and lipolytic activity, respectively. Anti-listerial activity is of most interest as it can contribute to reduce risk of listeriosis due to dairy products consumption, while proteolysis and lipolysis are responsible for some remarkable features (e. g. flavor, taste, and texture) of fermented foods. In summary, this study represents a contribution towards revealing some phenotypic and safety traits of a large set of strains of Enterococcus strains through the use of multivariate statistics combined with high throughput tools.

Short communication: Potential use of passion fruit (Passiflora cincinnata) as a biopreservative in the production of coalho cheese, a traditional Brazilian cheese

Passion fruit (Passiflora cincinnata Mast.) is a native fruit from the Caatinga, a typical ecoregion in northeastern Brazil, and it has potential for use by the food and pharmaceutical industries. In this study, we characterized the antimicrobial activity of P. cincinnata and its application in the production of coalho cheese, a traditional Brazilian product. Aqueous extract of P. cincinnata exhibited high inhibitory activity against Listeria spp. (n = 4, reference strains), Staphylococcus aureus (n = 3, reference strains), and multidrug-resistant Staph. aureus (n = 8), and low inhibitory activity against lactic acid bacteria (LAB, n = 3, reference strains). Based on these results, we produced coalho cheese using goat milk with and without (control) passion fruit. Cheeses were stored at 10°C for 14 d and populations of mesophilic aerobes, Staph. aureus, and presumptive LAB were monitored at d 1, 7 and 14. The passion fruit cheese had lower counts of mesophilic aerobes, Staph. aureus (after 7 and 14 d), and presumptive LAB (after 14 d) than the control cheese. Adding ground passion fruit contributed to a reduction of Staph. aureus counts in goat cheese, although these differences were not significant. These results indicated the inhibitory potential of passion fruit and its potential use for controlling microbial populations in a cheese model; further studies are needed to characterize the active molecules that are responsible for such activity.