RATIONALE: Filaggrin (FLG) gene mutation carriers have increased risk of atopic dermatitis (AD). ... more RATIONALE: Filaggrin (FLG) gene mutation carriers have increased risk of atopic dermatitis (AD). Herein we describe the clinical features of Japanese children with AD with and without FLG-null mutations. METHODS: Children with moderate-to-severe AD whose parents consented to filaggrin gene mutation analysis during February 2009 to July 2014 were enrolled. Each child was genotyped for the eight most prevalent FLG-null mutations in the Japanese population (R501X,
Introdução: A dermatite atópica (DA) é uma doença cutânea crónica imunologicamente mediada em que... more Introdução: A dermatite atópica (DA) é uma doença cutânea crónica imunologicamente mediada em que a maioria dos doentes está colonizada por Staphylococcus aureus (S. aureus) capaz de produzir vários factores de virulência. O S. aureus pode ser cultivado em cerca de 90% das lesões cutâneas e pode colonizar a pele de aspecto morfologicamente normal. Os estafilococos coagulase negativos (SCN) geralmente não produzem toxinas com actividade superantigénica, mas o seu papel patogénico na DA não pode ser excluído. Neste estudo, pretende-se caracterizar a comunidade estafilocócica da pele de doentes com DA e indivíduos saudáveis, assim como identificar factores de virulência nas espécies identificadas. Métodos: Todos os isolados estafilocócicos foram submetidos a análise numérica de factores de virulência. As espécies isoladas da pele de doentes com DA e indivíduos saudáveis foram submetidas a técnicas de identificação molecular por Multiplex-PCR para identificação de bactérias pertencentes...
Oral candidiasis is particularly evident, not only in cancer patients receiving chemotherapy, but... more Oral candidiasis is particularly evident, not only in cancer patients receiving chemotherapy, but also in elderly people with xerostomy. In general, Candida is an opportunistic pathogen, causing infections in immunocompromised people and, in some cases, when the natural microbiota is altered. Chitosan, a natural derivative of chitin, is a polysaccharide that has been proven to possess a broad spectrum of antimicrobial activity that encompasses action against fungi, yeast and bacteria. While recent studies have revealed a significant antibiofilm activity upon several microorganisms, including C. albicans, little is known regarding the impact of chitosan upon the adhesive process or mature biofilms. With that in mind, the purpose of this work was to evaluate, in vitro, the capability of chitosan to inhibit C. albicans growth and biofilm formation. The results obtained showed that chitosan is capable of inhibiting C. albicans planktonic growth (HMW, 1 mg/mL; LMW, 3 mg/mL). Regarding biofilm growth, chitosan inhibited C. albicans adhesion (ca. 95%), biofilm formation (percentages above 90%) and reduced mature biofilms by ca. 65% and dual species biofilms (C. albicans and S. mutans) by ca. 70%. These results display the potential of this molecule to be used as an effective anti-Candida agent capable of acting upon C. albicans infections.
World journal of microbiology & biotechnology, Jan 13, 2018
Over the years the body of work relating chitosan and its possible skin-related applications has ... more Over the years the body of work relating chitosan and its possible skin-related applications has grown, showing that chitosan is active both as a free compound and as a functional finishing of textiles. As a free molecule chitosan proved itself to be an attractive option as it is biocompatible and has a demonstrated biological activity (e.g. growth inhibition or adhesion inhibition) upon multiple skin pathogens, even upon multidrug resistant species. Furthermore, chitosan has wound healing accelerating properties, which make it a prime candidate for possible control of skin related infections. Almost inconspicuously, textiles have been one the main lines of defense of the skin against foreign threats, acting as a physical barrier to microbial colonization and infection. With the advent of textile functionalization specially designed textiles with enhanced protective characteristics, such as antimicrobial capacity, have come to the forefront. Chitosan functionalized textiles have bee...
Nowadays, nosocomial skin infections are increasingly harder to manage and control. In the search... more Nowadays, nosocomial skin infections are increasingly harder to manage and control. In the search for new, natural compounds capable of being alternatives to traditional antibiotics, chitosan and its nanoparticles, have garnered attention. This work sought to understand the potential of chitosan NPs in the management of infections caused by MDR skin pathogens in planktonic and sessile assays. Additionally, NPs' capacity to inhibit biofilm quorum sensing and prevent HaCat infections was also evaluated. The results obtained showed that chitosan NPs had an average size and charge of 226.6 ± 5.24 nm and +27.1 ± 3.09 mV. Inhibitory and bactericidal concentrations varied between 1 and 2 mg/mL and 2-7 mg/mL, respectively. Chitosan NPs effectively inhibited biofilm growth for all microorganisms and possessed strong anti-quorum sensing activity. Lastly, chitosan NPs proved to be effective interfere with A. baumannii's infection of HaCat cells, as they significantly reduced intracellu...
Annals of Applied Microbiology & Biotechnology Journal
Os dermatófitos são um grupo de fungos que conseguem invadir as superfícies queratinizadas de hum... more Os dermatófitos são um grupo de fungos que conseguem invadir as superfícies queratinizadas de humanos e outros animais e origenar uma infeção -dermatofitose. Estas infeções são um importante problema de saúde pública e, para as controlar, é necessário terapia prolongada. No entanto, os medicamentos existentes parecem exibir efeitos secundários e o uso frequente e prolongado destes compostos é responsável pela existência de estirpes resistentes a antifúngicos, o que representa um risco potencial para o ambiente e saúde humana. Por isso, são necessárias novas drogas biocompatíveis para usos prolongados. O quitosano é um polissacarídeo catiónico e biocompatível que possui atividade antimicrobiana. Assim, o principal objetivo deste trabalho foi avaliar a atividade antifúngica do quitosano sobre alguns dermatófitos e algumas espécies de Aspergillus sp. Para isso, a concentração mínima inibitória dos quitosanos foi determinada e os resultados mostraram que o quitosano possui atividade antifúngica contra T. rubrum e M. canis, apresentando CMIs que variam entre 1.1 e 2.2 mg/mL. Para as espécies de Aspergillus, não foi possível determinar as CMIs do quitosano. A concentração mínima fungicida também foi obtida para os dermatófitos, apresentando os mesmos valores obtidos para as CMIs. Como os dermatófitos são responsáveis por infeções das superfícies queratinizadas, é preciso compreender se o quitosano exerce algum efeito na atividade fúngica. A análise de imagens de microscopia eletrónica de varimento mostou que o quitosano parece ter um efeito protetor do substrato usado, o cabelo, quando este foi infetado com dermatófitos. O estudo do efeito do quitosano na atividade enzimática usando protease K revelou uma atividade importante na prevenção da ação das proteases. O efeito do quitosano na degradação de queratina por M. canis e T. rubrum também foi estudado pelo teste da "keratinazure" e os resultados indicaram que a libertação de cor era menor quando o quitosano estava presente no meio de cultura. A análise microscópica da superfície da "keratin-azure" quando o quitosano estava presente no meio de cultura corroborou as conclusões anteriores porque a superfície da "keratin-azure" tratada com quitosano mostrou-se intacta, apesar da existência de estruturas fúngicas à sua volta. Com base nestes resultados, é possível concluir que o quitosano apresenta uma atividade antifúngica relevante contra os dermatófitos, mostrando-se uma alternativa promissora aos tratamentos comuns para a tinha. i Abstract Dermatophytes are a goup of fungi that can invade keratinized tissues of humans and other animals and produce an infection -dermatophytosis. Dermatophytic infections are an important public health problem and to control them a prolonged therapy is required. But, the existing drugs seem to exhibit side effects and the frequent and prolonged use of these compounds is responsible for strain resistance, representing a potential risk for the environment and human health. Because of this features, new drug biocompatible formulations for long term use are required. Chitosan is a cationic, biocompatible polysaccharide that possess antimicrobial action. So, the main objective of this work was to evaluate the antifungal activity of chitosan upon some dermatophytes (T. rubrum and M. canis ) and some Aspergillus (A. terreus, A. niger, A. flavus and A. fumigatus). For this, the MIC of chitosans was determined and the results showed that chitosan possesses antifungal activity against T. rubrum and M. canis, presenting MIC ranging from 1.1 to 2.2 mg/mL. For Aspergillus species, in the range of tested concentrations it was not possible to determine the chitosan's MICs. Minimum fungicidal concentrations were also obtained for both dermatophytes, corresponding in both cases to the values obtained for MIC. As dermatophytes are responsible for infections of keratinized surfaces, is crucial to understand if chitosan exert any effect on fungi activity. The analysis of SEM images showed that chitosan seems to have a protective effect of the hair (substract), when this was infected with dermatophytes.
Vancomycin-resistant microorganisms are a hurdle that traditional antibiotics struggle to overcom... more Vancomycin-resistant microorganisms are a hurdle that traditional antibiotics struggle to overcome. These difficulties have led to search for new solutions based on natural products. Chitosan has been recognized as an effective antibacterial agent against a vast array of microorganisms including antibiotic resistant ones. As such, this work aimed to evaluate chitosan as an alternative to traditional antibiotics in the management/control of two vancomycin-resistant microorganisms, VRSA and VREF, in planktonic and sessile settings. The results obtained showed that chitosan was highly effective in inhibiting VRSA and VREF planktonic growth and reduced VREF viable counts by 6 log CFU in 30min. Additionally, chitosan was active upon several phases of VRSA and VREF sessile growth inhibiting adhesion, biofilm formation and dual-species biofilms at concentrations as low as 0.0125mg/mL. In lieu of these results chitosan shows great potential as a possible alternative for the control of vanco...
Quinoa and amaranth belong to the group of the so called “superfoods” and have a nutritional comp... more Quinoa and amaranth belong to the group of the so called “superfoods” and have a nutritional composition that confers multiple benefits.
Bioactive food packaging is an innovative approach for the prevention of the growth of food-spoil... more Bioactive food packaging is an innovative approach for the prevention of the growth of food-spoilage microorganisms.
Antimicrobial finishing of textiles has been found to be an economical way to prevent (or treat) ... more Antimicrobial finishing of textiles has been found to be an economical way to prevent (or treat) skin disorders. Hence, this research effort was aimed at elucidating the relationship between molecular weight (MW) of chitosan and its antimicrobial activity upon six dermal reference microorganisms, as well as the influence of the interactions with cotton fabrics on said activity. Using 3 chitosans with different MW, as well as two chitooligosaccharide (COS) mixtures, a relevant antimicrobial effect was observed by 24 h for the six microorganisms tested; it was apparent that the antimicrobial effect is strongly dependent on the type of target microorganism and on the MW of chitosan being higher for lower MW in the case of E. coli, K. pneumoniae and P. aeruginosa, and the reverse in the case of both Gram-positive bacteria. Furthermore, a strong anti-fungal effect was detectable upon C. albicans, resembling the action over Gram-positive bacteria. Interactions with cotton fabric resulted in a loss of COS activity when compared with cultured media, relative to the effect over Gram-negative bacteria. However, no significant differences for the efficacy of all the 5 compounds were observed by 4 h. The three chitosans possessed a higher antimicrobial activity when impregnated onto the fabric, and presented a similar effect on both Gram-positive bacteria and yeast, in either matrix. Pseudomonas aeruginosa showed to be the most resistant microorganism to all five compounds.
International Journal of Polymeric Materials and Polymeric Biomaterials, 2015
In the last years, the number of products for oral care has been expanded to adapt to consumer ne... more In the last years, the number of products for oral care has been expanded to adapt to consumer needs. Thus, in addition to conventional products, new ''pocket products'' such as sugar-free chewing gums and oral strips (OS) have been developed for oral care. In the present study, OS were formulated using chitosan as the film-forming polymer in adequate concentrations to also be used as antimicrobial agent. Other strip components, such as the type of plasticizer, were also optimized. Mechanical properties of the optimal OS were evaluated and, due to chitosan's characteristic astringency, the strips were also sensorially evaluated.
Traditional manufacture of bread from maize has been noted to play important roles from both econ... more Traditional manufacture of bread from maize has been noted to play important roles from both economic and social standpoints; however, enforcement of increasingly strict hygiene standards requires thorough knowledge of the adventitious microbiota of the departing dough. To this goal, sourdough as well as maize and rye flours from several geographic locations and in two different periods within the agricultural year were assayed for their microbiota in sequential steps of quantification and identification. More than 400 strains were isolated and taxonomic differentiation between them was via Biomerieux API galleries (375 of which were successfully identified) following preliminary biochemical and morphological screening. The dominant groups were yeasts and lactic acid bacteria (LAB). The most frequently isolated yeasts were Saccharomyces cerevisiae and Candida pelliculosa. The most frequently isolated LAB were (heterofermentative) Leuconostoc spp. and (homofermentative) Lactobacillus...
RATIONALE: Filaggrin (FLG) gene mutation carriers have increased risk of atopic dermatitis (AD). ... more RATIONALE: Filaggrin (FLG) gene mutation carriers have increased risk of atopic dermatitis (AD). Herein we describe the clinical features of Japanese children with AD with and without FLG-null mutations. METHODS: Children with moderate-to-severe AD whose parents consented to filaggrin gene mutation analysis during February 2009 to July 2014 were enrolled. Each child was genotyped for the eight most prevalent FLG-null mutations in the Japanese population (R501X,
Introdução: A dermatite atópica (DA) é uma doença cutânea crónica imunologicamente mediada em que... more Introdução: A dermatite atópica (DA) é uma doença cutânea crónica imunologicamente mediada em que a maioria dos doentes está colonizada por Staphylococcus aureus (S. aureus) capaz de produzir vários factores de virulência. O S. aureus pode ser cultivado em cerca de 90% das lesões cutâneas e pode colonizar a pele de aspecto morfologicamente normal. Os estafilococos coagulase negativos (SCN) geralmente não produzem toxinas com actividade superantigénica, mas o seu papel patogénico na DA não pode ser excluído. Neste estudo, pretende-se caracterizar a comunidade estafilocócica da pele de doentes com DA e indivíduos saudáveis, assim como identificar factores de virulência nas espécies identificadas. Métodos: Todos os isolados estafilocócicos foram submetidos a análise numérica de factores de virulência. As espécies isoladas da pele de doentes com DA e indivíduos saudáveis foram submetidas a técnicas de identificação molecular por Multiplex-PCR para identificação de bactérias pertencentes...
Oral candidiasis is particularly evident, not only in cancer patients receiving chemotherapy, but... more Oral candidiasis is particularly evident, not only in cancer patients receiving chemotherapy, but also in elderly people with xerostomy. In general, Candida is an opportunistic pathogen, causing infections in immunocompromised people and, in some cases, when the natural microbiota is altered. Chitosan, a natural derivative of chitin, is a polysaccharide that has been proven to possess a broad spectrum of antimicrobial activity that encompasses action against fungi, yeast and bacteria. While recent studies have revealed a significant antibiofilm activity upon several microorganisms, including C. albicans, little is known regarding the impact of chitosan upon the adhesive process or mature biofilms. With that in mind, the purpose of this work was to evaluate, in vitro, the capability of chitosan to inhibit C. albicans growth and biofilm formation. The results obtained showed that chitosan is capable of inhibiting C. albicans planktonic growth (HMW, 1 mg/mL; LMW, 3 mg/mL). Regarding biofilm growth, chitosan inhibited C. albicans adhesion (ca. 95%), biofilm formation (percentages above 90%) and reduced mature biofilms by ca. 65% and dual species biofilms (C. albicans and S. mutans) by ca. 70%. These results display the potential of this molecule to be used as an effective anti-Candida agent capable of acting upon C. albicans infections.
World journal of microbiology & biotechnology, Jan 13, 2018
Over the years the body of work relating chitosan and its possible skin-related applications has ... more Over the years the body of work relating chitosan and its possible skin-related applications has grown, showing that chitosan is active both as a free compound and as a functional finishing of textiles. As a free molecule chitosan proved itself to be an attractive option as it is biocompatible and has a demonstrated biological activity (e.g. growth inhibition or adhesion inhibition) upon multiple skin pathogens, even upon multidrug resistant species. Furthermore, chitosan has wound healing accelerating properties, which make it a prime candidate for possible control of skin related infections. Almost inconspicuously, textiles have been one the main lines of defense of the skin against foreign threats, acting as a physical barrier to microbial colonization and infection. With the advent of textile functionalization specially designed textiles with enhanced protective characteristics, such as antimicrobial capacity, have come to the forefront. Chitosan functionalized textiles have bee...
Nowadays, nosocomial skin infections are increasingly harder to manage and control. In the search... more Nowadays, nosocomial skin infections are increasingly harder to manage and control. In the search for new, natural compounds capable of being alternatives to traditional antibiotics, chitosan and its nanoparticles, have garnered attention. This work sought to understand the potential of chitosan NPs in the management of infections caused by MDR skin pathogens in planktonic and sessile assays. Additionally, NPs' capacity to inhibit biofilm quorum sensing and prevent HaCat infections was also evaluated. The results obtained showed that chitosan NPs had an average size and charge of 226.6 ± 5.24 nm and +27.1 ± 3.09 mV. Inhibitory and bactericidal concentrations varied between 1 and 2 mg/mL and 2-7 mg/mL, respectively. Chitosan NPs effectively inhibited biofilm growth for all microorganisms and possessed strong anti-quorum sensing activity. Lastly, chitosan NPs proved to be effective interfere with A. baumannii's infection of HaCat cells, as they significantly reduced intracellu...
Annals of Applied Microbiology & Biotechnology Journal
Os dermatófitos são um grupo de fungos que conseguem invadir as superfícies queratinizadas de hum... more Os dermatófitos são um grupo de fungos que conseguem invadir as superfícies queratinizadas de humanos e outros animais e origenar uma infeção -dermatofitose. Estas infeções são um importante problema de saúde pública e, para as controlar, é necessário terapia prolongada. No entanto, os medicamentos existentes parecem exibir efeitos secundários e o uso frequente e prolongado destes compostos é responsável pela existência de estirpes resistentes a antifúngicos, o que representa um risco potencial para o ambiente e saúde humana. Por isso, são necessárias novas drogas biocompatíveis para usos prolongados. O quitosano é um polissacarídeo catiónico e biocompatível que possui atividade antimicrobiana. Assim, o principal objetivo deste trabalho foi avaliar a atividade antifúngica do quitosano sobre alguns dermatófitos e algumas espécies de Aspergillus sp. Para isso, a concentração mínima inibitória dos quitosanos foi determinada e os resultados mostraram que o quitosano possui atividade antifúngica contra T. rubrum e M. canis, apresentando CMIs que variam entre 1.1 e 2.2 mg/mL. Para as espécies de Aspergillus, não foi possível determinar as CMIs do quitosano. A concentração mínima fungicida também foi obtida para os dermatófitos, apresentando os mesmos valores obtidos para as CMIs. Como os dermatófitos são responsáveis por infeções das superfícies queratinizadas, é preciso compreender se o quitosano exerce algum efeito na atividade fúngica. A análise de imagens de microscopia eletrónica de varimento mostou que o quitosano parece ter um efeito protetor do substrato usado, o cabelo, quando este foi infetado com dermatófitos. O estudo do efeito do quitosano na atividade enzimática usando protease K revelou uma atividade importante na prevenção da ação das proteases. O efeito do quitosano na degradação de queratina por M. canis e T. rubrum também foi estudado pelo teste da "keratinazure" e os resultados indicaram que a libertação de cor era menor quando o quitosano estava presente no meio de cultura. A análise microscópica da superfície da "keratin-azure" quando o quitosano estava presente no meio de cultura corroborou as conclusões anteriores porque a superfície da "keratin-azure" tratada com quitosano mostrou-se intacta, apesar da existência de estruturas fúngicas à sua volta. Com base nestes resultados, é possível concluir que o quitosano apresenta uma atividade antifúngica relevante contra os dermatófitos, mostrando-se uma alternativa promissora aos tratamentos comuns para a tinha. i Abstract Dermatophytes are a goup of fungi that can invade keratinized tissues of humans and other animals and produce an infection -dermatophytosis. Dermatophytic infections are an important public health problem and to control them a prolonged therapy is required. But, the existing drugs seem to exhibit side effects and the frequent and prolonged use of these compounds is responsible for strain resistance, representing a potential risk for the environment and human health. Because of this features, new drug biocompatible formulations for long term use are required. Chitosan is a cationic, biocompatible polysaccharide that possess antimicrobial action. So, the main objective of this work was to evaluate the antifungal activity of chitosan upon some dermatophytes (T. rubrum and M. canis ) and some Aspergillus (A. terreus, A. niger, A. flavus and A. fumigatus). For this, the MIC of chitosans was determined and the results showed that chitosan possesses antifungal activity against T. rubrum and M. canis, presenting MIC ranging from 1.1 to 2.2 mg/mL. For Aspergillus species, in the range of tested concentrations it was not possible to determine the chitosan's MICs. Minimum fungicidal concentrations were also obtained for both dermatophytes, corresponding in both cases to the values obtained for MIC. As dermatophytes are responsible for infections of keratinized surfaces, is crucial to understand if chitosan exert any effect on fungi activity. The analysis of SEM images showed that chitosan seems to have a protective effect of the hair (substract), when this was infected with dermatophytes.
Vancomycin-resistant microorganisms are a hurdle that traditional antibiotics struggle to overcom... more Vancomycin-resistant microorganisms are a hurdle that traditional antibiotics struggle to overcome. These difficulties have led to search for new solutions based on natural products. Chitosan has been recognized as an effective antibacterial agent against a vast array of microorganisms including antibiotic resistant ones. As such, this work aimed to evaluate chitosan as an alternative to traditional antibiotics in the management/control of two vancomycin-resistant microorganisms, VRSA and VREF, in planktonic and sessile settings. The results obtained showed that chitosan was highly effective in inhibiting VRSA and VREF planktonic growth and reduced VREF viable counts by 6 log CFU in 30min. Additionally, chitosan was active upon several phases of VRSA and VREF sessile growth inhibiting adhesion, biofilm formation and dual-species biofilms at concentrations as low as 0.0125mg/mL. In lieu of these results chitosan shows great potential as a possible alternative for the control of vanco...
Quinoa and amaranth belong to the group of the so called “superfoods” and have a nutritional comp... more Quinoa and amaranth belong to the group of the so called “superfoods” and have a nutritional composition that confers multiple benefits.
Bioactive food packaging is an innovative approach for the prevention of the growth of food-spoil... more Bioactive food packaging is an innovative approach for the prevention of the growth of food-spoilage microorganisms.
Antimicrobial finishing of textiles has been found to be an economical way to prevent (or treat) ... more Antimicrobial finishing of textiles has been found to be an economical way to prevent (or treat) skin disorders. Hence, this research effort was aimed at elucidating the relationship between molecular weight (MW) of chitosan and its antimicrobial activity upon six dermal reference microorganisms, as well as the influence of the interactions with cotton fabrics on said activity. Using 3 chitosans with different MW, as well as two chitooligosaccharide (COS) mixtures, a relevant antimicrobial effect was observed by 24 h for the six microorganisms tested; it was apparent that the antimicrobial effect is strongly dependent on the type of target microorganism and on the MW of chitosan being higher for lower MW in the case of E. coli, K. pneumoniae and P. aeruginosa, and the reverse in the case of both Gram-positive bacteria. Furthermore, a strong anti-fungal effect was detectable upon C. albicans, resembling the action over Gram-positive bacteria. Interactions with cotton fabric resulted in a loss of COS activity when compared with cultured media, relative to the effect over Gram-negative bacteria. However, no significant differences for the efficacy of all the 5 compounds were observed by 4 h. The three chitosans possessed a higher antimicrobial activity when impregnated onto the fabric, and presented a similar effect on both Gram-positive bacteria and yeast, in either matrix. Pseudomonas aeruginosa showed to be the most resistant microorganism to all five compounds.
International Journal of Polymeric Materials and Polymeric Biomaterials, 2015
In the last years, the number of products for oral care has been expanded to adapt to consumer ne... more In the last years, the number of products for oral care has been expanded to adapt to consumer needs. Thus, in addition to conventional products, new ''pocket products'' such as sugar-free chewing gums and oral strips (OS) have been developed for oral care. In the present study, OS were formulated using chitosan as the film-forming polymer in adequate concentrations to also be used as antimicrobial agent. Other strip components, such as the type of plasticizer, were also optimized. Mechanical properties of the optimal OS were evaluated and, due to chitosan's characteristic astringency, the strips were also sensorially evaluated.
Traditional manufacture of bread from maize has been noted to play important roles from both econ... more Traditional manufacture of bread from maize has been noted to play important roles from both economic and social standpoints; however, enforcement of increasingly strict hygiene standards requires thorough knowledge of the adventitious microbiota of the departing dough. To this goal, sourdough as well as maize and rye flours from several geographic locations and in two different periods within the agricultural year were assayed for their microbiota in sequential steps of quantification and identification. More than 400 strains were isolated and taxonomic differentiation between them was via Biomerieux API galleries (375 of which were successfully identified) following preliminary biochemical and morphological screening. The dominant groups were yeasts and lactic acid bacteria (LAB). The most frequently isolated yeasts were Saccharomyces cerevisiae and Candida pelliculosa. The most frequently isolated LAB were (heterofermentative) Leuconostoc spp. and (homofermentative) Lactobacillus...
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