Pseudomonas carnis isolated from blue discolored fresh cheese and insights into the phylogeny.

Most Pseudomonas spp. are responsible for spoilage in refrigerated foods such as alteration in flavor, texture and appearance. Samples of Minas Frescal cheese with blue discoloration were analysed and contained a high Pseudomonas concentration (7.72 ± 0.36 log CFU/g). Out of the 26 Pseudomonas isolates that were analyzed in our study, 19 demonstrated the capability of producing a diffusible dark pigment. Thus, a pigment-producing isolate (C020) was selected by rep-PCR fingerprinting and subsequently subjected to whole-genome sequencing. The draft genome assembled comprises 42 contigs totaling 6,366,75 bp with an average G + C content of 59.97%, and the species prediction performed by TYGS server, based on the draft genome sequence, identified the C020 as Pseudomonas carnis. In order to investigate the phylogenetic relationships of this isolate with strains already identified of this species, we performed an analysis based on whole-genomic sequences. First, an analysis of all P. carnis genomes deposited in GenBank to date shows that 11% (4/37) are misidentified, and belong to the Pseudomonas paracarnis species. A comparative analysis based on phylogenomic analysis has showed that there is no evolutionary relationship between P. carnis strains carrying second copies of trp genes related to blue discoloration (trpABCDF). This finding reinforces the assertion that these genes are contained in a mobile genetic element. However, it is worth noting that all strains carrying these secondary gene copies have exclusively been isolated from food sources. This observation provides valuable insights into the potential origins and dispersion dynamics of this genetic trait within the species.

Effects of calcium chloride substitution on the physicochemical properties of Minas Frescal cheese.

The aim in this research paper was to investigate the effect of using calcium monophosphate (MCP) and MCP mixed with commercial phosphates salts, in total or partial replacement of calcium chloride (CaCl2) in the manufacture of Minas Frescal cheese. Initially, model cheeses were made to perform the rheological analysis during the coagulation process. Of these, the five best treatments were chosen to carry out the production of Minas Frescal cheese, used only CaCl2 and MCP, and partial replacements of MCP + polyphosphate, MCP + potassium monophosphate (MKP) and MCP. The cheeses showed no significant difference in physicochemical composition, yield and syneresis, however, the cheese with partial replacement of CaCl2 by MCP + polyphosphate and MCP + MKP showed the highest hardness values, like the control. This demonstrates that it is possible to replace calcium chloride without significant changes in the physicochemical characteristics and yield of Minas Frescal cheese, and it is still possible to modulate the hardness of the cheese produced according to the type of calcium/phosphate source used. This allows the industry to replace the source of calcium in the manufacture of Minas Frescal cheese according to the desired hardness.

Consumers' attitudes of high-intensity ultrasound in Minas Frescal cheese processing: An innovative approach with text highlighting technique.

Due to the intense competition in the sector, the dairy market maintains a constant search for innovations. Thus, new technologies are incorporated, and new products are constantly launched, increasing the range of consumer options. In this way, the understanding of consumers' motivations, attitudes, and behaviors in the moments of choice, purchase, and consumption are important for the academic public and food industries. This study used the Text Highlighting methodology to assess Brazilian consumers' explicit attitudes towards using high-intensity ultrasound technology in Minas Frescal cheese processing. In the task, consumers were asked to highlight in a text the terms they "liked" or "disliked" about using high-intensity ultrasound in the Minas Frescal cheese processing. A seven-point Likert scale was also used to assess consumers' attitudinal statements. A high engagement of consumers with the Text Highlighting methodology could be observed (43.8-92.3% of text highlighting), which suggests good intuitiveness of the technique. Including information about the benefits of the emerging technology in the product, mainly on sensory and nutritional properties, may increase positive consumer perception, as it promotes the consumers to express their value judgment in the form of "liked". At the same time, the harms of the traditional processing technique prompted consumers to express their value judgment in the "disliked" highlights. It was observed that information should be in a simple and direct language, as technical terminology in the text did not have a positive effect. The categorizing of consumers according to the sentimental score showed that consumers are still reticent to use emerging technologies in Minas Frescal cheese processing. Consumers' attitudinal statements demonstrated that consumers perceive high-intensity ultrasound as a positive idea and safety technology for Minas Frescal cheese processing. Still, they are not willing to pay premium prices. In conclusion, Text Highlighting produced valuable insights that can be used in communication strategies with Minas Frescal cheese consumers.

Thermosonication as a pretreatment of raw milk for Minas frescal cheese production.

Minas frescal cheese is extremely popular in Brazil, with high perishability and acceptability. Among emerging technologies, ultrasound stands out for its satisfactory results regarding microbiological safety and technological and sensory aspects. The combined mild temperature application, called thermosonication, can generate even more promising results. In this study, a high-intensity ultrasound system combined with thermal heating (TS, thermosonication) was applied for the treatment of raw milk to produce Minas Frescal cheese. US energy was delivered to raw milk samples using a probe operating at a 20 kHz of frequency and nominal power of 160, 400, and 640 W. The TS system was compared with conventional pasteurization (HTST, high-temperature short-time pasteurization) at 72 to 75 °C and 15 s. Soft cheeses were prepared with different samples: (a) raw milk (control), b)conventionally pasteurized milk (HTST), and c) TS treat milk in different nominal power (TS160, TS400, and TS640). The produced cheeses were evaluated for microbiological behavior, rheology, color parameters, and bioactive compounds. TS treatment in milk resulted in higher microbial inactivation and stability during storage, improved color parameters (higher lightness (L*), and whiteness index (WI). TS treatment also showed a higher generation of bioactive compounds (higher antioxidant, and inhibitory activities of α-amylase, α-glucosidase, and angiotensin-converting enzymes) than HTST. The impact of TS on rheological properties was similar to HTST, resulting in more brittle and less firm products than the cheese produced with raw milk. The positive effects were more prominent using a nominal power of 400 W (TS400). Therefore, TS proved to be a promising process for processing milk for Minas Frescal cheese production.

A New Bacteriocin from Latilactobacillus sakei: In vitro and In situ Application.

AIMS AND BACKGROUND: Natural preservatives are a viable alternative to replace chemical preservatives that have potential toxicity and carcinogenic effects. OBJECTIVE: To prove the effectiveness in increasing the microbiological stability of Minas Frescal cheese with the addition of a bacteriocin obtained from Latilactobacillus sakei as a natural preservative. METHODS: A new broad-spectrum bacteriocin was evaluated for its functional activity in vitro and in situ when applied in the formulation of Minas Frescal cheese. A commercial bacteriocin was used as a positive control. RESULTS: The inhibitory action of the bacteriocin studied was confirmed, with a reduction of 42.86% in the count of coagulase-positive Staphylococcus in relation to the negative control, at the end of the 30 days of study. For the group of thermotolerant coliforms, the bacteriocin studied showed greater efficiency than the commercial preservative. In vitro analyzes showed the inhibitory action of bacteriocin, above 87% inhibition against S. aureus, E. coli and Salmonela enteritidis, and approximately 90% against Listeria monocytogenes. CONCLUSION: It was concluded that the bacteriocin produced by the Latilactobacillus sakei strain has great potential for application in foods such as Minas Frescal cheese.

Licury Cake in Diets for Lactating Goats: Qualitative Aspects of Milk and Cheese.

The study was carried out to evaluate the effects of licury cake (LC) inclusion in diets for lactating goats on milk chemical composition and fatty acid profile, and physicochemical composition and sensory attributes of Minas Frescal cheese. Twelve lactating goats were used (eight Saanen and four Anglo Nubian goats, with 35 ± 5 days in lactation and average body weight of 37.93 ± 9.22 kg), distributed in a triplicate 4 × 4 Latin Square design. The increasing levels of LC studied were: 0, 67, 133 and 200 g/kg of dry matter (DM). The analyses were adjusted using orthogonal polynomial contrasts, in which the probability level of 5% was considered. For sensory attributes, principal component analysis was performed. The LC inclusion promoted the reduction in moisture (p = 0.013) and mineral matter (p < 0.001) content in cheese. The ether extract content in cheese showed a quadratic effect (p = 0.021). Lauric acid showed a linear (p = 0.002) increasing effect, and myristic (p < 0.001) and rumenic (p = 0.018) acids showed quadratic effects. The sensory attributes analysis in cheese indicated that the inclusion of 133 and 200 g/kg of LC improve its texture. Flavor was improved with the inclusion of 67 and 133 g/kg. The inclusion of up to 133 g/kg of LC in the diet of lactating goats is recommended because it promotes improvements in flavor and texture of Minas Frescal cheese.

Palm Kernel Cake in Diets for Lactating Goats: Qualitative Aspects of Milk and Cheese.

We investigated the effect of including palm kernel cake (PKC) at the levels of 0, 80, 160 and 240 g kg-1 in the diet of lactating goats on the quality and sensory parameters of Minas Frescal cheese. Twelve goats were used in a triple 4 × 4 Latin square design. The dietary addition of PKC was associated with a reduction in moisture (p = 0.004), which compromised the cheese yield (p = 0.030). The ether extract content showed a quadratic response. There was a decrease in caproic (p = 0.014), caprylic (p = 0.011), capric (p = 0.003) and palmitic (p = 0.049) acids and an increase in lauric (p = 0.012) and myristic (p = 0.02) acids. Monounsaturated fatty acids increased (p = 0.008), whereas the ratio of polyunsaturated to saturated fatty acids (p = 0.022) and thrombogenicity index (p = 0.023) decreased. Principal component analysis indicated that control treatment and the treatment with 80 g kg-1 PKC were the most acceptable to the cheese tasters. The use of palm kernel cake is a good alternative for lactating goats when added to the diet at levels up to 80 g kg-1.

New Active Packaging Based on Biopolymeric Mixture Added with Bacteriocin as Active Compound.

The objective of this work was to develop a chitosan/agar-agar bioplastic film incorporated with bacteriocin that presents active potential when used as food packaging. The formulation of the film solution was determined from an experimental design, through the optimization using the desirability function. After establishing the concentrations of the biopolymers and the plasticizer, the purified bacteriocin extract of Lactobacillus sakei was added, which acts as an antibacterial agent. The films were characterized through physical, chemical, mechanical, barrier, and microbiological analyses. The mechanical properties and water vapor permeability were not altered by the addition of the extract. The swelling property decreased with the addition of the extract and the solubility increased, however, the film remained intact when in contact with the food, thus allowing an efficient barrier. Visible light protection was improved by increased opacity and antibacterial capacity was effective. When used as Minas Frescal cream cheese packaging, it contributed to the increase of microbiological stability, showing a reduction of 2.62 log UFC/g, contributing a gradual release of the active compound into the food during the storage time. The film had an active capacity that could be used as a barrier to the food, allowing it to be safely packaged.

Presence and growth prediction of Staphylococcus spp. and Staphylococcus aureus in Minas Frescal cheese, a soft fresh cheese produced in Brazil.

Physical-chemical characteristics of Minas Frescal cheese (MFC) favor the growth of Staphylococcus spp. and allow the production of enterotoxins by specific strains. Here, we aimed to characterize the physical-chemical aspects (pH, storage temperature, and salt content) and the presence of Staphylococcus spp. in MFC samples (n = 50) to support a modeling study for the growth by this microorganism. Coagulase-positive staphylococci isolates were obtained and subjected to PCR assays to identify them as Staphylococcus aureus (nuc) and to detect staphylococcal enterotoxin-related genes (sea, seb, sec, sed, see). Staphylococcus aureus growth kinetics (maximum growth rate, Grmax, and lag time) were predicted based on ComBase model and MFC physical-chemical aspects. Mean counts of Staphylococcus spp. ranged from 3.3 to 6.7 log cfu/g, indicating poor hygiene practices during production. Selected isolates (n = 10) were identified as S. aureus, but none presented classical enterotoxin-related genes. pH, temperature, and salt content ranged from 5.80 to 6.62, 5°C to 12°C, and 0.85% to 1.70%, respectively. The Grmax values ranged from 0.012 to 0.419 log cfu/g per h. Independent of the storage temperature, the lowest Grmax values (0.012 to 0.372 log cfu/h) were obtained at pH 5.80 associated with salt content of 1.7%; independent of the pH and salt content, the best temperature to avoid staphylococcal growth was 7.5°C. Hygienic conditions during MFC production must be adopted to avoid staphylococcal contamination, and storage at temperatures lower than 7.5°C can prevent staphylococcal growth and the potential production of enterotoxins.

Shiga toxin-producing Escherichia coli isolated from pasteurized dairy products from Bahia, Brazil.

The presence of pathogenic Shiga toxin-producing Escherichia coli (STEC) in dairy products represents a public health concern because of its ability to produce the toxins Stx1 and Stx2, which cause intestinal diseases. Monitoring the stages of milk production and checking dairy products for contamination are crucial steps to ensure dairy safety. This study aimed to report the occurrence of thermotolerant coliforms, E. coli, and STEC strains in pasteurized dairy products and to evaluate the antibiotic resistance profiles, serotypes, and characterizations of the STEC isolates by pulsed-field gel electrophoresis. We obtained a total of 138 pasteurized dairy products from 15 processing plants in Bahia, Brazil, to examine coliforms, E. coli, and STEC strains. We found that 43% of samples (59/138) contained thermotolerant coliforms, and 30% (42/138) did not comply with Brazilian regulations. Overall, 6% (9/138) were positive for E. coli and 4% (5/138) were positive for STEC. We recovered 9 STEC isolates from pasteurized cream (2/9), Minas Padrão cheese (2/9), Minas Frescal cheese (4/9), and ricotta (1/9). All isolates were stx2-positive, and 2 were eae-positive. All isolates were negative for the "big 6" STEC serogroups, belonging instead to serotypes ONT:HNT, ONT:H12, O148:H-, OR:H40, OR:HNT, and O148:HNT. Pulsed-field gel electrophoresis revealed 100% genetic similarity among 3 isolates from 2 different samples produced in the same production facility, which may suggest cross-contamination. As well, we found isolates that were 98% similar but in samples produced in different production facilities, suggesting a mutual source of contamination or a circulating strain. Two STEC strains exhibited resistance to streptomycin. Although the isolates presented a low resistance profile and no strain belonged to the "big 6" pathogenic group, the circulation of stx2-positive STEC strains in ready-to-eat products highlights the importance of epidemiological surveillance inside the Brazilian dairy chain.

Therapeutic Effects of Probiotic Minas Frescal Cheese on the Attenuation of Ulcerative Colitis in a Murine Model.

Inflammatory bowel diseases (IBDs) constitute disturbances of gastrointestinal tract that cause irreversible changes in the structure and function of tissues. Ulcerative colitis (UC), the most frequent IBD in the population, is characterized by prominent inflammation of the human colon. Functional foods containing probiotic bacteria have been studied as adjuvants to the treatment or prevention of IBDs. The selected probiotic strain Lactococcus lactis NCDO 2118 (L. lactis NCDO 2118) exhibits immunomodulatory effects, with promising results in UC mouse model induced by dextran sodium sulfate (DSS). Additionally, cheese is a dairy food that presents high nutritional value, besides being a good delivery system that can be used to improve survival and enhance the therapeutic effects of probiotic bacteria in the host. Therefore, this work investigated the probiotic therapeutic effects of an experimental Minas Frescal cheese containing L. lactis NCDO 2118 in DSS-induced colitis in mice. During colitis induction, mice that consumed the probiotic cheese exhibited reduced in the severity of colitis, with attenuated weight loss, lower disease activity index, limited shortening of the colon length, and reduced histopathological score. Moreover, probiotic cheese administration increased gene expression of tight junctions' proteins zo-1, zo-2, ocln, and cln-1 in the colon and increase IL-10 release in the spleen and lymph nodes. In this way, this work demonstrates that consumption of probiotic Minas Frescal cheese, containing L. lactis NCDO 2118, prevents the inflammatory process during DSS-induced colitis in mice, opening perspectives for the development of new probiotic functional foods for personalized nutrition in the context of IBD.

Effect of probiotic Minas Frescal cheese on the volatile compound and metabolic profiles assessed by nuclear magnetic resonance spectroscopy and chemometric tools.

This study aimed to evaluate the effect of Lacticaseibacillus casei 01 as a probiotic culture on the production of volatile organic compounds and metabolic profile of Minas Frescal cheese. Lactose (α-lactose and ß-lactose), fatty acids (unsaturated and saturated), citric acid, tryptophan, and benzoic acid were the main compounds. Compared with the control cheese, probiotic cheese was characterized by the highest concentration of tryptophan and presented a higher number of volatile acids. The control cheese was characterized by the highest concentration of benzoic acid and fatty acids, resulting in a higher number of volatile alcohols and esters. No differences were observed for α-lactose, ß-lactose, and citric acid contents. A clear separation of probiotic and control Minas Frescal cheese was obtained using 1H nuclear magnetic resonance spectra, demonstrating that the addition of probiotic culture altered the metabolic profile of Minas Frescal cheese. Overall, the findings suggested that the addition of probiotic culture promoted the proteolysis in the fresh cheeses, decreased the lipolysis, and altered the volatile compounds. Furthermore, nuclear magnetic resonance spectroscopy coupled to chemometrics tools could be used to differentiate probiotic and conventional cheeses.

Antimicrobial resistance of Staphylococcus spp. isolated from organic and conventional Minas Frescal cheese producers in São Paulo, Brazil.

The genus Staphylococcus is recognized worldwide as a cause of bacterial infections in humans and animals. Antibiotics used in dairy cattle combined with ineffective control can increase antimicrobial resistance. The objective of this study was to characterize 95 Staphylococcus strains isolated from organic and conventional Minas Frescal cheese production regarding antibiotic resistance (phenotype and genotype), presence of sanitizer-resistant genes and biofilm-formation genes, and SCCmec typing. Most strains (25.3%) showed higher resistance to penicillin, followed by oxacillin (21.1%) and clindamycin (11.6%). Among antibiotic resistance genes, the most prevalent were blaZ (25.3%), mecA (13.7%), lsaB (6.3%), msrA (4.2%), ant4 (3.2%), and tetM (2.1%); among sanitizer-resistance genes they were qacA/B (5.3%) and qacC (6.3%); and among biofilm, bap (4.2%), icaA (29.5%), icaD (41.1%). However, there was no statistically significant difference between organic and conventional dairy products, possibly due to the lack of synthetic antibiotic use on conventional farms during the sample collection period. Methicillin-resistant Staphylococcus aureus (MRSA) had their SCCmec identified as types I and IVc, and the methicillin-resistant coagulase-negative staphylococci had nontypeable SCCmec. These results suggest that there are antibiotic-resistant strains in both organic and conventional Minas Frescal cheese production in the state of São Paulo, Brazil. This supports the idea that improved quality control is needed from the milking stage up to the final product.

Milk-clotting proteases from Pleurotus albidus: an innovative alternative for the production of Minas frescal cheese

Pleurotus albidus, a naturally growing species in the Amazon region, has been considered a promising source of milk-clotting proteases. The production of such enzymes using lignocellulosic residues is a sustainable alternative to replace mammalian rennet. The application of P. albidus milk-clotting proteases in cheese making has not yet been reported in the scientific literature. The aim of this study was to characterize the milk-clotting proteases of P. albidus and use these enzymes in the production of Minas frescal cheese. For the production of coagulating proteases, the mushroom was grown in açaí seeds supplemented with rice bran (10%, w/w). The parameters affecting the production of coagulant, such as inoculum size, fermentation time, initial pH of cultivation medium and age of the inoculum were evaluated. The coagulant extract obtained under optimal production conditions was evaluated for optimal pH and temperature, pH and temperature stability, effect of ions and inhibitors. Significant production of coagulating proteases was obtained under the following conditions: inoculum size (2.5%), fermentation time (10 days), initial pH of the cultivation medium (6), and inoculum age (10 days). The coagulant exhibited significant catalytic activity in pH 5.0 at 55°C, with stability at 45°C and was completely inhibited by iodoacetic acid. The milk-clotting proteases of P. albidus were efficient for making Minas frescal cheese that presented 55.0% of moisture, 20.0% of lipids and 17.20% of protein. Pleurotus albidus is a potential source of milk-clotting proteases that can be applied in dairy industry for production of fresh Minas frescal cheese.

Milk-clotting proteases from Pleurotus albidus: an innovative alternative for the production of Minas frescal cheese

Pleurotus albidus, a naturally growing species in the Amazon region, has been considered a promising source of milk-clotting proteases. The production of such enzymes using lignocellulosic residues is a sustainable alternative to replace mammalian rennet. The application of P. albidus milk-clotting proteases in cheese making has not yet been reported in the scientific literature. The aim of this study was to characterize the milk-clotting proteases of P. albidus and use these enzymes in the production of Minas frescal cheese. For the production of coagulating proteases, the mushroom was grown in açaí seeds supplemented with rice bran (10%, w/w). The parameters affecting the production of coagulant, such as inoculum size, fermentation time, initial pH of cultivation medium and age of the inoculum were evaluated. The coagulant extract obtained under optimal production conditions was evaluated for optimal pH and temperature, pH and temperature stability, effect of ions and inhibitors. Significant production of coagulating proteases was obtained under the following conditions: inoculum size (2.5%), fermentation time (10 days), initial pH of the cultivation medium (6), and inoculum age (10 days). The coagulant exhibited significant catalytic activity in pH 5.0 at 55°C, with stability at 45°C and was completely inhibited by iodoacetic acid. The milk-clotting proteases of P. albidus were efficient for making Minas frescal cheese that presented 55.0% of moisture, 20.0% of lipids and 17.20% of protein. Pleurotus albidus is a potential source of milk-clotting proteases that can be applied in dairy industry for production of fresh Minas frescal cheese.(AU)

Resistome in gram-negative bacteria from soft cheese in Brazil / Resistoma em bactérias gram-negativas de queijo minas frescal no Brazil

Objective: evaluation of antibiotic resistance in Gram-negative microbiota from ready-to-eat cheese samples. Methodology: this research applied an adapted methodology to select from a food sample viable Gram-negative microbiota displaying antibiotic resistance. The selected food was a cheese that is commonly consumed without thermal processing, the Minas Frescal cheese. The evaluation was followed by a PCR screening in this resistant microbiota, for genes that provide resistance to antibiotics and also to the quaternary ammonium. Results: all cheese samples harbored a resistant microbiota. In 13.3% of the cheese samples analyzed, the resistance reached all ten different antibiotics tested and, in 80%, 8 to 10 different antibiotics. In antibiotics considered critics as the carbapenems: ertapenem presented resistant microbiota in 86.7% of the samples. In cephalosporins, the resistance reached 100% in the third generation (ceftazidime) and almost half of the samples (46.7%) in the fourth generation (cefepime). In genotypic research, seven different resistance genes were found in 69.2% of the bacterial pools, including the beta-lactamase-producing genes ctx, tem, shv, tetracycline-resistant genes, and a high rate of integrons class 1 and 2. Conclusion: the results indicate phenotypically and genotypically that the Minas Frescal cheese can harbor potential resistant microbiota. Therefore, the methodology used is a viable possibility and with a broader answer about the food microbiota role in resistance. This research corroborates the food area as an important sector to be managed to reduce the process of antibiotic resistance.

Objetivo: avaliação da resistência a antibióticos em microbiota Gram-negativa de amostras de queijo prontas para consumo. Metodologia: esta pesquisa aplicou uma metodologia adaptada para selecionar a microbiota Gram-negativa viável apresentando resistência a antibióticos em uma amostra de alimento. O alimento selecionado foi um queijo frequentemente consumido sem processamento térmico, o queijo Minas Frescal. A avaliação foi seguida de uma triagem por PCR, nesta microbiota resistente, para genes que fornecem resistência aos antibióticos e também ao quaternário de amônio. Resultados: todas as amostras de queijo apresentaram microbiota resistente. Em 13,3% dos queijos analisados essa resistência alcançou todos os 10 diferentes antibióticos testados e em 80% entre 8 e 10 antibióticos diferentes. Em antibióticos considerados críticos como os carbapenêmicos: ertapenem apresentou microbiota resistente em 86,7% das amostras. Nas cefalosporinas, a resistência atingiu 100% na terceira geração (ceftazidima) e quase a metade das amostras (46,7%) na quarta geração (cefepime). Na pesquisa genotípica, sete diferentes genes de resistência foram encontrados em 69,2% dos pools bacterianos, incluindo o genes produtores de beta-lactamase, genes de resistência à tetraciclina, ctx, tem, shv e uma alta taxa de integron classe 1 e 2. Conclusão: os resultados indicam fenotipicamente e genotipicamente que o queijo Minas Frescal pode apresentar uma potencial microbiota resistente. Portanto, a metodologia utilizada é uma possibilidade viável e com uma resposta mais ampla sobre o papel da microbiota na resistência. Esta pesquisa corrobora a área de alimentos como um setor importante a ser gerenciado para redução no processo de resistência a antibióticos.

Monensin residues in the production of Minas Frescal cheese: Stability, effects on fermentation, fate and physicochemical characteristics of the cheese.

Considering the widespread use of the antibiotic monensin (MON) in the Brazilian livestock and the possibility of residues in milk, this paper aimed to study the stability and fate of this drug during the production of Brazilian Minas Frescal cheese, its effects on milk fermentation and on the physicochemical characteristics of this product. For that, samples of raw milk were fortified with MON at three different nominal concentrations (1.0, 2.0 and 8.0 µg/kg), passed through heat treatment and used to produce Minas Frescal cheese. Pasteurization efficiency was certified by alkaline phosphatase and peroxidase enzyme tests and cheese samples were evaluated for pH, moisture and total protein and fat content. MON residues were determined by LC-MS/MS in the following steps: raw milk, heat-treated milk, whey and cheese. No significant degradation of MON due to heat treatment was observed, suggesting that the drug is resistant to high temperatures. Moreover, the residue levels quantified in cheese and whey demonstrated a concentration of this antibiotic in the curd by about 5-fold, with a small amount of MON being lost during draining. There were no significant differences (p > 0.05) considering the physicochemical parameters evaluated in cheese samples. Fermentation was also not affected by the presence of the drug. The results showed that residues of MON in milk are stable during cheese production and may be concentrated in the final product, as well as indicate the need to establish a MON safe residue level for this food commodity.

Sensory acceptance and physico-chemical composition of mixed minas frescal cheese / Aceitação sensorial e composição físico-química de queijo minas frescal misto

Buffalo milk presents higher proteins and lipids concentration than cow milk, which provides a better yield in cheese-making production and products with considerable sensory approval. However, buffalo milk is not regularly available throughout the year due to different handling conditions. Thus, to guarantee the supply of buffalo milk dairy products during the year, the addition of bovine milk is an alternative. Therefore, this study aimed to test the effect of a buffalo and bovine milk mixture for the Minas Frescal cheese elaboration by physico-chemical, sensory analysis and obtaining yield. The raw material was analysed for the parameters of acidity, density, fat, total solids and solid-not-fat. There were 3 Minas Frescal cheeses elaborated from 3 formulations: 100% buffalo milk, 100% bovine milk and the mixture of 50% of each milk. The cheeses were submitted to the Gravimetric and Gerber methods to obtain values of moisture and fat, as well as to obtain, indirectly, fat in dry matter. For bovine and buffalo milk, the parameters evaluated (acidity, density, fat, total solids and solids-not-fat) complied with Brazilian legislation and parameters described in the literature. For the bovine, mixed and buffalo fresh cheeses, values were obtained, respectively, for moisture (74.04, 60.93 and 63.61), fat in dry matter (44.35, 42.23 and 46.03) and cheese yield (27, 20.8 and 24.2), indicating a higher yield for the bovine Minas Frescal cheese and higher fat content for the buffalo cheese. The overall acceptance of the mixed Minas Frescal cheese was significantly superior to the bovine and buffalo cheese. The parameters of colour, appearance, texture, flavour and overall acceptance were above 8 points in the hedonic scale ('moderately liked' to 'extremely liked'), and the aroma attribute scored was above 7 ('I enjoyed regularly' to 'moderately liked'). Therefore, the elaboration of Minas Frescal cheese from the mixture of 2 matrices (buffalo and bovine milk) demonstrated technological viability with the potential to meet the demands of the consumer market.

O leite de búfala possui maior porcentagem de todos os componentes tornando-o uma matéria-prima adequada com potencial para fornecer um maior rendimento e qualidade nutricional na produção de queijo. No entanto, o leite de búfalo não está disponível regularmente ao longo do ano devido a diferentes condições de manejo. Assim, para garantir a oferta de produtos lácteos com leite de búfala independentemente da época, a adição de leite bovino pode ser vista como uma alternativa. Logo, este trabalho teve como objetivo testar o efeito da mistura do leite de búfalo e bovino na elaboração do queijo Minas Frescal por meio de análises físico-químicas e sensoriais e obtenção do rendimento economico. A matéria-prima foi analisada quanto aos parâmetros: acidez titulável, densidade, gordura, sólidos totais e sólidos totais desengordurados. Três variedades de queijo Minas Frescal foram elaboradas a partir de três formulações: 100% de leite de búfala, 100% de leite bovino e da mistura de 50% de cada leite. Os queijos foram submetidos aos métodos Gavimetric e Gerber para obtenção de umidade e gordura, bem como, indiretamente, para obtenção de matéria gorda no extrato seco. Para o leite bovino e de búfala, os parâmetros avaliados (acidez titulável, densidade, gordura, sólidos totais e sólidos totais desengordurados) estavam de acordo com a legislação brasileira e parâmetros descritos na literatura. Para os queijos bovino, misto e de búfala, foram obtidos, respectivamente, para: umidade (74,04; 60,93 e 63,61), gordura na matéria seca (44,35; 42,23 e 46,03) e rendimento econômico (27, 20,8 e 24,2) indicando maior rendimento o queijo Minas Frescal bovino e maior teor de gordura para o queijo búfalo. A aceitação global do queijo Minas Frescal misto foi significativamente superior ao do queijo bovino e bubalino. Os parâmetros de cor, aparência, textura, sabor e aceitabilidade global foram acima de 8 pontos (entre "gostei moderadamente" e "extremamente gostei") e o atributo aroma obteve pontuação acima de 7 ("gostei regularmente" para "gostei moderadamente"). Portanto, a elaboração do queijo Minas Frescal da mistura de leite bovino ao búfalo demonstrou potencial para atender as demandas do mercado consumidor.

Effect of Lactobacillus rhamnosus on growth of Listeria monocytogenes and Staphylococcus aureus in a probiotic Minas Frescal cheese.

This study aimed to evaluate the effect of Lactobacillus rhamnosus GG on growth of Staphylococcus aureus and Listeria monocytogenes, inoculated alone or in combination on surface of Minas Frescal cheeses, during storage for 21 days at 7 °C. Survival percentages of each individual bacterial species after exposure to in vitro simulated gastrointestinal conditions (SGC) were also determined. The addition of L. rhamnosus did not affect (P > 0.05) pH, moisture, fat, protein and texture profile of Minas Frescal cheeses. L. rhamnosus was able to survive in suitable counts (>6 Log CFU/g) in cheeses from the 7th day of storage, with high survival (>74.6-86.4%) after SGC. An inhibitory effect of L. rhamnosus on L. monocytogenes was observed in cheeses (decrease of 1.1-1.6 Log CFU/g) and after SGC (20% reduction in the survival). No inhibitory effect of L. rhamnosus was observed on S. aureus counts (P > 0.05), and this microorganism did not survive the exposure to SGC. In conclusion, the addition of L. rhamnosus in Minas Frescal cheese has a potential for L. monocytogenes inhibition. Further studies are necessary to elucidate the mechanisms involved in the inhibition process and determine the survival ability of the bacterial species evaluated in in vivo experiments.

Ohmic heating does not influence the biochemical properties of Minas Frescal cheese but decreases uric acid levels in healthy Wistar rats.

The objective of this study was to evaluate the effects of the consumption of Minas Frescal cheese (2 g for 15 d) produced with pasteurized milk or milk subjected to ohmic heating on the health variables (biochemical, hematological, and uric acid levels) of rats, as well as on the identity variables and proteolysis indices of cheese. Three groups of rats (n = 6 per group) were studied, as follows: control group, without cheese consumption (CON), rats fed Minas Frescal cheese produced with milk pasteurized by conventional technology (PC, 72 to 75°C for 15 s), and rats fed Minas Frescal cheese produced with milk subjected to ohmic heating (OHC, 8 V/cm, 72 to 75°C for 15 s). Ohmic heating did not alter the macronutrient levels (moisture, fat, protein) of the cheeses, but an increase in the proteolysis index was observed. For both cheeses, no significant differences were observed for feed intake (139 to 143 g/d), water consumption (196 to 206 mL/d), or final animal weight (236 to 252 g), and no changes were detected in hematological variables (counts of red blood cells, hemoglobin, leukocytes, and platelets) or levels of potassium, calcium, magnesium, urea, creatinine, total cholesterol, high-density lipoprotein cholesterol, blood glucose, and bilirubin (total, direct, and indirect) of rats. In addition, both cheeses led to a reduction of serum triglycerides and an increase in monocyte, eosinophil, and basophil levels. However, consumption of OHC resulted in decreased sodium, uric acid, and lymphocyte levels. The present results suggest that ohmic heating in the processing of Minas Frescal cheese has positive effects on the health of healthy rats.