Abstract The purpose of this review was to describe how adaptive laboratory evolution (ALE) can provide improvement to lactic acid bacteria (LAB) strains for their application in industrial biotechnological processes. This review was carried out according to the Preferred Reporting Items for Systematic Review and Meta-analysis (PRISMA) approach, incorporating the ScienceDirect and Scopus databases. The literature search yielded 4,167 (ScienceDirect) and 27 (Scopus) articles, which were reduced to 12 after applying the inclusion /exclusion criteria. The studies revolved around LAB of the genera Lactobacillus, Lactococcus, Leuconostoc and Enterococcus and the application of ALE experiments in batch mode, fed-batch mode, or both, and aimed to produce strains with increased lactic acid production capabilities, higher cell viability, and multiple-stress tolerance. The studies demonstrated that ALE is an efficient approach for strain modification towards desired phenotypic functions and does not require genetic engineering. Knowledge of the cellular and molecular responses of microorganisms to stress enables an understanding of the adaptation mechanisms of LAB strains for survival and increased production of metabolites throughout ALE experiments.

Resumo O objetivo deste artigo de revisão foi descrever como a adaptação laboratorial evolutiva (ALE) pode proporcionar o melhoramento de cepas de bactérias ácido-lácticas (BAL) para aplicação em processos biotecnológicos industriais. A revisão foi realizada de acordo com a abordagem Preferred Reporting Items for Systematic Review and Meta-analysis (PRISMA), empregando as bases de dados ScienceDirect e Scopus. A busca bibliográfica resultou em 4.167 e 27 documentos, respectivamente, os quais reduziram-se para 12 após a aplicação dos critérios de inclusão/exclusão. Os trabalhos utilizaram BAL dos gêneros Lactobacillus, Lactococcus, Leuconostoc e Enterococcus para a ALE, empregando o método batelada ou batelada alimentada ou ambos, e tinham como objetivo a geração de cepas com maior capacidade de produção de ácido láctico, viabilidade celular e tolerância a diferentes condições de estresse. Os estudos demonstraram que a ALE é uma ferramenta eficiente no melhoramento de cepas para os fenótipos desejados, além de não empregar engenharia genética. O conhecimento das respostas celulares e moleculares dos microrganismos ao estresse possibilita a compreensão dos mecanismos de adaptação das cepas de BAL para sobrevivência e maior produção de metabólitos ao longo da ALE.

Resumo As plantas alimentícias não convencionais têm uma variedade de compostos bioativos com valor nutricional. O jaracatiá (Vasconcellea quercifolia) pertence à família Caricaceae e é uma alternativa dietética com uma excelente composição nutricional. Este estudo teve como objetivo caracterizar a composição nutricional da farinha de frutos verdes do jaracatiá (V. quercifolia), a fim de aplicá-la em um alimento funcional. Para isso, a farinha foi caracterizada em relação a macro e micronutrientes, fatores antinutricionais, pH, atividade da água e cor. Esta farinha apresentou teores de carboidratos de 22,31%; proteínas de 9,65%; fibra alimentar dietética de 32,80%; lipídios de 14,95%, dos quais 63,56% são ácidos graxos insaturados, especialmente ácido oleico; e cinzas de 9,10%, com maiores concentrações para potássio, cálcio e magnésio. Portanto, a farinha de V. quercifolia tem características nutritivas e pode ser utilizada como suplemento alimentar.

Abstract Non-conventional food plants have a variety of bioactive compounds with nutritional value. Vasconcellea quercifolia A. St.-Hil., belonging to the Caricaceae family, is a dietary alternative with excellent nutritional composition. This study aimed at characterizing the nutritional composition of mountain papaya (V. quercifolia) green fruit flour, in order to incorporate it in a functional food. For that purpose, the flour was characterized regarding its macro and micronutrients, anti-nutritional factors, pH, water activity, and color. This flour showed contents of carbohydrate of 22.31%; protein of 9.65%; dietary fiber of 32.80%; lipids of 14.95%, 63.56% of which are unsaturated fatty acids, especially oleic acid; and ash of 9.10%, with higher concentrations for potassium, calcium and magnesium. Therefore, V. quercifolia flour had good nutritional characteristics and might be used as supplementary food.

Abstract Cheese whey is the main by-product obtained in the production of cheese. Despite its high nutritional value, approximately half of the whey volume generated is still disposed incorrectly, which causes damage to the ecosystem due to the high cheese whey pollutant load. Therefore, it is important to use this by-product and its components in an increasing number of applications, especially as food ingredient. This review aimed to show the technology of production of butter from whey cream, as well as showing the physico-chemical, sensory, and nutritional characteristics of the product. There were no significant variations in the physico-chemical composition of milk cream butter and whey cream butter in the literature available. As the technology to produce whey butter is quite simple, this by-product has potential to be exploited by the dairy industry. Additionally, further studies on production process, characterization, and sensory analysis are required to enable its large-scale production.

Resumo O soro de queijo é o principal coproduto obtido durante o processo de fabricação de queijos. Apesar de possuir um valor nutricional elevado, cerca da metade do volume de soro gerado ainda é descartada de forma incorreta, constituindo-se uma prática prejudicial para o ecossistema devido à enorme carga poluente do soro de queijo. Dessa forma, é importante a utilização desse coproduto e de seus componentes em um número maior de aplicações, especialmente como ingrediente alimentício. O objetivo deste artigo de revisão é apresentar a tecnologia para produção da manteiga a partir do reúso do creme de soro de queijo, demonstrando as características físico-químicas, sensoriais e nutricionais do produto, e apontando as alternativas e possíveis utilizações para os principais componentes do soro. Na comparação entre a manteiga de creme de leite e a manteiga de creme de soro, na literatura disponível, os resultados não apresentam variações significativas em relação à sua composição físico-química. Dada a simplicidade da tecnologia de produção, a manteiga de soro possui potencial para ser explorada pelas indústrias lácteas. Além disso, mais estudos sobre o processo de fabricação, caracterização e análise sensorial são necessários para viabilizar sua produção em maior escala.

Abstract This study aimed to make and characterize a fresh cheese using the surplus of bovine colostrum from dairy farms. The bovine colostrum was characterized in terms of fat (4.00%), protein (16.30%), moisture (79.68%), ash (0.95%), minerals, pH (6.32), titratable acidity (0.27 g/100 mL), immunoglobulin G (>50 g/L), lactose (1.60%), color, and the presence of pathogenic bacteria. The fresh cheese made with colostrum was characterized in the same terms as the bovine colostrum (fat (7.00%), protein (22.95%), moisture (67.98%), ash (1.85%), minerals, pH (6.15), titratable acidity (0.04 g/100 mL), immunoglobulin G (30.95 g/L), lactose (not detectable), color, and the presence of pathogenic bacteria), with the addition of the texture and sensory analysis. The colostrum cheese had high moisture content (67.98%) and low-fat content (7.00%). The protein content was also high (22.95%), whose major percentage was composed by immunoglobulins. An important finding in this work was associated with the possible absence of lactose in the colostrum cheese, thus opening opportunities for future research regarding the development of dairy products for lactose intolerant consumers. In addition, the high concentration of immunoglobulin G might give this product an immune boost feature. The sensorial analysis showed that the recipe of the fresh cheese requires improvement to achieve higher acceptance from the public, especially regarding texture.

Resumo Este trabalho teve como objetivo a elaboração e caracterização de um queijo fresco a partir do excedente de colostro bovino de fazendas leiteiras. O colostro foi caracterizado quanto aos teores de gordura (4,00%), proteínas (16,30%), umidade (79,68%), cinzas (0,95%), minerais, pH (6,32), acidez titulável (0,27 g/100 mL), imunoglobulina G (>50 g/L), lactose (1,60%), cor e presença de bactérias patogênicas. O queijo fresco elaborado com colostro foi submetido às mesmas caracterizações do colostro: gordura (7,00%), proteínas (22,95%), umidade (67,98%), cinzas (1,85%), minerais, pH (6,15), acidez titulável (0,04 g/100 mL), imunoglobulina G (30,95 g/L), lactose (não detectável), cor e presença de bactérias patogênicas, além da avaliação dos parâmetros de textura e da análise sensorial. O queijo fresco elaborado com colostro apresentou alto teor de umidade (67,98%) e baixo teor de gordura (7,00%). A porcentagem de proteína foi elevada (22,95%), sendo grande parte composta por imunoglobulinas. O teor de lactose do queijo fresco elaborado com colostro foi muito baixo para ser quantificado, indicando a possibilidade de desenvolvimento de novos produtos lácteos a partir de colostro para consumidores intolerantes à lactose. Além disso, o queijo apresentou elevada concentração de imunoglobulina G, que pode conferir a esse produto a propriedade de reforço imunológico. A análise sensorial mostrou que o processo de elaboração do queijo fresco com colostro deve ser aprimorado para aumentar a aceitação dos consumidores, principalmente no que se refere à textura.

Abstract The aim of the present study was to evaluate the viability of probiotic bacteria Lactobacillus acidophilus (LA-5)® and Bifidobacterium lactis (BB-12)® encapsulation using spray drying (SD) with ricotta whey supplemented with gelatin and hydrolyzed collagen as encapsulating material. Following previous experiments, these gelatin:collagen ratios were defined: 7:3 (gelatin:collagen) for L. acidophilus LA-5 and 9:1 for B. lactis BB-12, which had reduced bacterial counts of 0.46 and 1.26 log Colony-Forming Units (CFU)/g, respectively. There was a viability loss of 0.70 log CFU/g and 0.34 log CFU/g for LA-5 and BB-12, respectively, throughout storage (30 days, 25 ºC). When exposed to the simulated gastric juice, encapsulated LA-5 and BB-12 had higher survival rates (89.91 and 95.83%, respectively) than non-encapsulated microorganisms under the same conditions (54.78 and 57.27%, respectively). In the presence of bile salts, encapsulated LA-5 and BB-12 had survival of 77.44% and 51.05%, respectively, while free cells showed rates of over 99%. After 30 days, these rates decreased to 71.07 and 50.60%, for encapsulated LA-5 and BB-12, respectively. Therefore, ricotta whey associated to gelatin and hydrolyzed collagen can be considered a potential encapsulating agent for the SD of probiotic dairy bacteria.

Resumo O presente trabalho teve o objetivo de avaliar os efeitos da adição do bagaço de malte de cevada nas características de linguiças frescais de carne suína ao longo de 10 dias de armazenamento. Foram elaboradas quatro formulações de linguiça frescal, uma delas sem adição de bagaço de malte (formulação controle) e outras três com 3%, 6% e 9% do bagaço. As linguiças foram submetidas às análises físico-químicas para determinação dos teores de cinzas, umidade, proteínas, lipídeos, carboidratos, fibra bruta, atividade de água, pH, índice de peróxidos e valor energético. Também foram realizadas análises tecnológicas de textura e cor e avaliação sensorial. A linguiça com 3% de bagaço de malte apresentou no 10º dia de armazenamento teor proteico de 17,10% e teor de fibra bruta de 2,00%, enquanto na formulação controle esses teores foram de 16,59% e 0,77%, respectivamente, apresentando diferença significativa (p ≤ 0,05) entre as amostras, o que representa um aumento no valor nutricional do produto. Na análise de dureza, a formulação controle apresentou o valor de 13,99 N, e a formulação com 3% de bagaço, valor de 10,11 N, demonstrando que as linguiças com adição do bagaço ficaram menos rígidas. Na análise sensorial, a formulação controle apresentou cerca de 80% de aceitabilidade em todos os atributos; a linguiça com 3% de bagaço de malte atingiu índice de aceitação global superior a 80%; a linguiça com 6% de bagaço obteve índice de aproximadamente 70%; e a formulação com 9% apresentou índice de aceitação global de aproximadamente 64%. A adição de 3% de bagaço de malte na linguiça se mostrou a melhor alternativa, pois, além de proporcionar um aumento no valor nutricional do produto, apresentou uma boa aceitabilidade por parte dos consumidores. Os resultados obtidos neste estudo demonstram que o aproveitamento do bagaço de malte em produtos cárneos é uma alternativa viável, que contribui para a redução dos custos de produção e auxilia na solução de um problema ambiental.

Abstract The present study aimed to evaluate the effects of barley malt pomace addition on the characteristics of fresh pork sausages over 10 days of storage. Four fresh sausage formulations were elaborated, one with no addition of barley malt pomace (control formulation), and the other three elaborated with 3%, 6%, and 9% of pomace. The sausages were submitted to physicochemical analyses to determine ash, moisture, protein, lipid, carbohydrate, and raw fiber contents, water activity, pH, peroxide indices, and energetic value. Additionally, texture and color technological analysis and sensory analysis were performed. Sausages with 3% of malt pomace presented a protein content of 17.10% on the 10th day of storage, and raw fiber content of 2.00%; while in the control formulation these contents were 16.59% and 0.77%, respectively, with a significant difference (p ≤ 0.05) between samples, representing an increase in product nutritional value. In the hardness analysis, the control formulation had a value of 13.99 N and the formulation with 3% of pomace of 10.11 N, which shows that sausages with the addition of pomace are not as hard. In the sensory analysis, the control formulation showed about 80% acceptability in all attributes, the sausage with 3% of malt pomace had an acceptability index higher than 80% for the attribute 'global acceptance', the sausage with 6% of pomace had an index of approximately 70% and the formulation with 9% showed acceptability index of approximately 64% for the attribute 'global acceptance'. The addition of 3% of malt pomace to the sausage was the best alternative, since it had good acceptance by consumers, and provided an increased nutritional value. Our results show that the use of malt pomace in meat products is a viable alternative that helps to reduce production costs and aids in solving an environmental issue.

Resumo A doença celíaca é caracterizada pela incapacidade do organismo de digerir o glúten no intestino delgado, e o tratamento dietético consiste em evitar alimentos que contenham essa proteína. Casca de batata, semente de abóbora e quinoa são ingredientes que podem ser usados para produzir pão sem glúten. O objetivo deste trabalho foi avaliar as características químicas e sensoriais de pães isentos de glúten adicionados de farinhas de casca de batata (FCB), semente de abóbora (FSA) e quinoa (FQ). Foram realizadas a análise sensorial dos pães preparados sem glúten e a análise química das farinhas alternativas e das formulações de pães elaboradas. A formulação do pão com farinhas e amidos tradicionais que obteve maior aceitação na avaliação sensorial foi utilizada como base para preparar as três formulações de pães com 2,5%, 5,0% e 7,5% de cada uma das farinhas alternativas. Na análise química das farinhas, as concentrações de carboidratos da FCB e FQ não diferiram estatisticamente. Os níveis de lipídios, proteínas e cinzas da FSA foram significativamente maiores que os das demais farinhas. Os teores de carboidratos, cinzas e lipídios das três formulações elaboradas com as farinhas alternativas não apresentaram diferença significativa. A formulação com 5,0% das farinhas alternativas apresentou maior aceitação sensorial. Os resultados obtidos demonstram que as farinhas utilizadas são alternativas viáveis para a produção de pães sem glúten, o que torna possível melhorar a dieta da população com doença celíaca pelo aumento do teor nutricional de minerais, proteínas e lipídios.

Abstract Celiac disease is characterized by the body’s inability to digest gluten in the small intestine, and the dietary treatment consists of avoiding foods that contain this protein. Potato peel, pumpkin seeds, and quinoa are ingredients that can be used to produce gluten-free bread. The aim of this work was to evaluate chemical and sensory characteristics of gluten-free breads consisting of the idea to compare both made with traditional ingredients and with the addition of Potato Peel Flour (PPF), Pumpkin Seed Flour (PSF), and Quinoa Flour (QF). Sensory analysis of the prepared gluten-free breads and chemical analysis of the alternative flours were performed, as well as the formulations developed. The bread formulation with traditional flours and starches that obtained the highest acceptance in the sensory evaluation was used as a base to prepare three bread formulations, with 2.5%, 5.0%, and 7.5% of each alternative flours. Regarding the chemical analysis of the flours, carbohydrate concentrations of PPF and QF did not differ statistically. Lipid, protein, and ash levels in PSF were significantly higher than those in the other flours. Carbohydrate, ash, and lipid contents of the three formulations made with alternative flours showed no significant differences. The formulation with 5.0% of the alternative flours had higher sensory acceptance. The results showed that the alternative flours used are viable alternatives for the production of gluten-free breads, making it possible to improve the diet of part of the population with celiac disease by increasing the nutritional contents of minerals, lipids, and proteins.

Abstract We investigated the immobilization of a tetrameric Kluyveromyces lactis β-galactosidase (EC: 3.2.1.23) (KL-Gal) on Immobead 150 using different support modification strategies. Immobead support was modified using an acid solution of H2SO4:HNO3 (3:1) (Immobead-Ac) or 5 % (v/v) glutaraldehyde (Immobead-Glu). Its unmodified form (Immobead) was also tested. Immobilization yields and efficiencies were evaluated by testing protein loads from 10 to 200 mg.g-1 support. The thermal, physico-chemical, textural and catalytic properties of the supports (modified and unmodified) and their derivatives (Immobead-KL-Gal, Immobead-Ac-KL-Gal and Immobead-Glu-KL-Gal) were analyzed. The highest immobilization yields and efficiencies were achieved with a protein load of 100 mg.g-1 support. Surface and pore areas of the Immobead support were greatly decreased after modification. Michaelis constant of the immobilized β-galactosidase increased in the derivatives. Maximum velocity decreased approximately 2.8 times for Immobead-KL-Gal and Immobead-Glu-KL-Gal, and approximately 1.4 times for Immobead-Ac-KL-Gal. In batch processes, the three derivatives could be reused successfully at least 15 times, maintaining high residual enzymatic activity during the lactose hydrolysis (in both cheese whey and milk). The tetrameric K. lactis β-galactosidase immobilized on Immobead supports via the tested treatments was stabilized and is an alternative tool for lactose hydrolysis in the dairy industry.

Abstract Although cow milk is the most widely used dairy product, the milk of other animals presents characteristics relevant to the development of these products. Goat milk has higher digestibility and a lower incidence of allergic reactions than cow milk, while buffalo milk has higher levels of lipids, proteins and lactose. The objective of this work was to elaborate and evaluate the physicochemical, technological, microbiological and sensory characteristics of cream cheese formulations elaborated with different proportions of cow, goat and buffalo milk without whey removal. The milk mixture was allowed to produce cream cheese with high levels of ashes and proteins, and despite the retention of the whey, no occurrence of syneresis was observed in the formulations. The formulation elaborated with 33.33% of each type of milk obtained the highest positive intent to purchase, the highest acceptability index for overall impression and the highest score for the flavor attribute. Considering the number of probiotic microorganisms present in the cream cheese composition, the developed product can be considered a functional food. It is possible to prepare cream cheese with different types of milk and whey retention, creating a competitive product for the market and an alternative for the industry.

RESUMO: Um dos grandes desafios das indústrias de laticínios é destinar de forma correta todo o soro gerado durante a produção de queijo, devido ao seu impacto ambiental, grande volume gerado e potencial tecnológico. A hidrólise enzimática da lactose presente no soro de queijo é uma alternativa biotecnológica. Contudo, um dos fatores limitantes de sua utilização é o custo relativamente alto das enzimas, o que poderia ser minimizado com a imobilização destes biocatalisadores. Baseado nesse contexto, o objetivo do presente trabalho foi avaliar a enzima comercial β-galactosidase de Kluyveromyces lactis , imobilizada em esferas de alginato de cálcio e gelatina, empregando o glutaraldeído e a concanavalina A (ConA) como agentes modificadores, no processo de hidrólise da lactose presente no soro de queijo. Os resultados obtidos demonstraram que o encapsulamento da enzima complexada com ConA em esferas de alginato-gelatina, sem a presença de glutaraldeído no meio de imobilização, aumentou de modo significativo o teor de hidrólise da lactose, obtendo conversão máxima de 72%.

ABSTRACT: One of the greatest challenges for dairy industries is the correct destination of all the whey generated during cheese making, considering its high impact, the large volume created, and its technological potential. Enzymatic hydrolysis of cheese whey lactose is a biotechnological alternative. However, one of the limiting factors of its use is the relatively high cost of the enzymes, which could be lowered with the immobilization of these biocatalysts. Considering this context, the objective of this research was to evaluate the commercial Kluyveromyces lactis β-galactosidase enzyme immobilized in calcium alginate spheres and gelatin, using glutaraldehyde and concanavalin A (ConA) as modifying agents in the hydrolysis of cheese whey lactose process. Results have shown that the enzyme encapsulation complexed with ConA in alginate-gelatin spheres, without glutaraldehyde in the immobilization support, has significantly increased the hydrolysis of lactose rate, achieving a maximum conversion of 72%.

Background Strawberry is a pseudofruit mainly cultivated in temperate climate regions. Considering its high levels of ascorbic acid and phenolic compounds, the consumption of strawberry fruit can be beneficial to health. The Brazilian strawberry production revolves around 3000 tons per year, significantly influencing the food market and generating income to farmers. However, this production can be partially impaired by two-spotted spider mite (TSSM) Tetranychus urticae Koch infestations, due to decreases in the quality and quantity of fruit. Since there are no data in the literature about alterations caused by TSSM infestation in strawberry plants, our work aimed towards evaluating nutritional and physicochemical parameters of TSSM-infested strawberry plants, along with the related chemical treatment (CT) (acaricide) or biological treatment (predatory mite Phytoseiulus macropilis Banks). Results Strawberry fruit from TSSM-infested plants present the highest levels of acidity and exhibit low levels of anthocyanin and phenolic compounds, while fruit from TSSM-infested plants + biological control using predatory mite shows high levels of soluble solids, phenolic compounds and ascorbic acid, along with a high soluble solid content/titratable (SSC/TA) acidity ratio, which indicates high quality fruit. Conclusions Our results suggest that TSSM infestation decreases fruit quality and that the biological control of TSSM using a predatory mite is a suitable alternative to organic production, since the presence of predatory mite does not affect fruit quality and development.

Milkborne transmission of Shiga toxin- producing Escherichia coli (STEC) has raised considerable concern due to recent outbreaks worldwide and poses a threat to public health. The aim of this study was to develop a sensitive and specific multiplex PCR assay to detect the presence of STEC in bovine raw milk. To identify E. coli (ATCC 25922) contamination, the gene uspA was used, and PCR sensitivity and specificity were accessed by testing diluted samples ranging from 2 to 2.0 × 10(6) CFU/mL. To detect STEC, the stx1 and stx2 genes were selected as targets. After reaction standardization, the multiplex assay was tested in raw milk collected from 101 cows on dairy farms. PCR assay for E. coli detection had a specificity of 100% and sensitivity of 79% (P<0.0001), with a lower detection limit of 2 CFU/mL. Multiplex PCR assay had 100% sensitivity for E. coli positive raw milk samples, and 31.1% were contaminated with STEC, 28.3% of stx2, and 1.9% of stx1. The multiplex PCR assay described in the present study can be employed to identify and screen E. coli harboring stx1 and stx2 genes in raw milk on dairy farms and in industries.

As variações da microbiota de seis bateladas do queijo Serrano (3 maturadas no verão e 3 maturadas no inverno) foram estudadas durante a produção e maturação, com o objetivo de caracterizá-lo microbiologicamente. Este queijo é tradicionalmente elaborado com leite cru, sem a adição de cultura lática, por produtores rurais do sul do Brasil. Vários grupos microbianos foram enumerados no leite, na coalhada e em diferentes estágios ao longo dos 60 dias de maturação. Também foram investigados alguns parâmetros físico-químicos, tais como pH, acidez titulável, atividade de água, umidade e teor de NaCl. Determinaram-se os coeficientes de correlação entre os valores desses parâmetros e as contagens dos microrganismos ao longo da maturação. A influência da estação do ano também foi estudada. Os resultados demonstraram que a microbiota e as características físico-químicas do queijo Serrano apresentaram variações significativas durante o período de maturação de 60 dias, principalmente no verão. A maioria dos grupos microbianos atingiram suas contagens máximas no queijo aos 7 dias, diminuindo progressivamente até o final da maturação. O aumento nas contagens durante a primeira semana de maturação foi acompanhado por um declínio acentuado no valor do pH, devido à produção de ácido pelos microrganismos. As bactérias láticas constituíram-se no principal grupo microbiano do queijo Serrano e suas contagens foram similares as de microrganismos viáveis totais em todos os pontos amostrados. O grande número de lactobacilos durante a elaboração e maturação do queijo sugere que estes microrganismos possam ter um papel importante na produção deste queijo. Em conclusão, o trabalho aponta a importância de um tempo de maturação superior a trinta dias como fator de estabilização microbiológica deste produto artesanal, bem como o fato de ser um produto bastante homogêneo quanto às suas características físico-químicas e microbiológicas, independente da estação do ano, exceto para as contagens de halotolerantes e de bolores e do teor de umidade, que apresentaram diferenças significativas entre o verão e o inverno.

Changes in the microbial flora present in six Serrano cheese batches (3 ripened in Summer and 3 in Winter) were studied during production and ripening, to microbiologically characterize this cheese, which is traditionally manufactured by farmers in the south of Brazil, using raw cow milk, without the addition of starter cultures. The levels of several microbial groups were enumerated in the milk, curd and different stages of the 60-day long ripening period. Values of some physicochemical parameters, such as pH, titratable acidity, water activity, moisture and NaCl content were also determined. The correlation coefficients between these parameters and the microbial groups throughout ripening were determined. The influence of the season of the year was also studied. Results demonstrated that the microflora and the physicochemical characteristics of Serrano cheese changed significantly during the 60-day ripening period, especially in Summer. Almost all microbial groups reached their highest counts after the 7th day of maturation, steadily decreasing towards the end of the process. Higher microbial counts during the first week of ripening was accompanied by lactic acid production and, as a consequence, a sharp drop in the pH of cheese was observed. Lactic acid bacteria comprised the main microbial group present in Serrano cheese and their counts were similar to the total viable counts at all sampling points. The abundance of lactobacilli during the manufacture and ripening suggests that these microorganisms may play an important role in the production of Serrano cheese. In conclusion, this work shows the importance of ripening time being longer than 30 days to allow for microbiological stabilization of this artisanal product. Also, except for halotolerants and moulds counts, and for moisture content, there were no significant differences observed either for microbial counting or for physicochemical characteristics of the cheese concerning the season of the year in which the product was manufactured.