RESUMO O objetivo deste estudo foi avaliar o uso do biossensor de Maldi-Tof MS na avaliação microbiana de grãos de kefir. Maldi-Tof MS é uma nova metodologia para o diagnóstico rápido de micro-organismos. Um total de 358 micro-organismos foram isolados, 31 foram leveduras e 327 foram bactérias (divididas em bactérias láticas e acéticas). Colônias microbianas foram cultivadas em meio Luria-Bertani (LB) e incubadas a 35 °C por 18h e posteriormente utilizadas na identificação a nível de espécies pelo Maldi-Tof MS. A população microbiana identificada nos grãos de kefir brasileiros foi Lactobacillus paracasei, Saccharomyces cerevisiae, Lactobacillus plantarum, Acetobacter pasteurianus e Acetobacter syzygii. Este estudo demonstrou uma identificação rápida e precisa dos micro-organismos de grãos de kefir brasileiros utilizando o biossensor Maldi-Tof MS. Em conclusão, a tecnologia Maldi-Tof MS pode facilitar o controle microbiológico em um processo de fermentação usando grãos de kefir como culturas iniciadoras.

ABSTRACT The aim of this study was to evaluate the use of Maldi-Tof MS biosensor in microbial assessment of Brazilian kefir grains. Maldi-Tof MS is a new methodology for the rapid diagnosis of microorganisms. A total of 358 microorganisms were isolated, 31 were yeasts and 327 were bacteria (divided into lactic and acetic bacteria). Microbial colonies were grown in Luria-Bertani agar medium and incubated at 35 °C for 18h and used in the identification of species by Maldi-Tof MS. The microbial population identified in Brazilian kefir grains was Lactobacillus paracasei, Saccharomyces cerevisiae, Lactobacillus plantarum, Acetobacter pasteurianus, and Acetobacter syzygii. This study demonstrated a rapid and accurate identification of the Brazilian kefir grains microorganisms using the Maldi-Tof MS biosensor. In conclusion, the Maldi-Tof MS technology can facilitate the microbiological control in a fermentation process using kefir grains as starter cultures.

Resumo: O objetivo deste trabalho foi verificar o uso de biossensor a laser para a avaliação da atividade metabólica microbiana do vinagre de kefir. Os grãos de kefir foram inoculados em mosto de maçã e analisados diariamente pela técnica de biospeckle. A atividade biológica inicial foi de 14,21 pixels por valor absoluto das diferenças (AVD). A atividade biológica dos grãos de kefir diminuiu nos primeiros dias de fermentação e aumentou no quarto dia para 11,51 pixels por AVD; no entanto, no último dia, a atividade biológica dos grãos de kefir diminuiu para 7,12 pixels (fermentação alcoólica) e 6,77 pixels por AVD (fermentação acética). Esta nova metodologia do biospeckle facilita o controle microbiológico em processos fermentativos.

Abstract: The objective of this work was to verify the use of a laser biosensor for the microbial metabolic activity assessment of kefir vinegar. Kefir grains were inoculated in apple must and were analyzed daily using the biospeckle technique. The initial biological activity was of 14.21 pixels per absolute value of differences (AVD). The biological activity of kefir grains decreased in the early days of fermentation and increased on the fourth day to 11.51 pixels per AVD; however, on the last day, the biological activity of kefir grains decreased to 7.12 pixels (alcoholic fermentation) and to 6.77 pixels per AVD (acetic fermentation). This new biospeckle methodology facilitates the microbiological control in fermentation processes.

Abstract The aim of this study was to develop a kefir apple-based vinegar and evaluate this fermentation process using new methodology with Biospeckle Laser. Brazilian kefir grains were inoculated in apple must for vinegar production. In this study, the microbial community present in kefir, and correspondent vinegar, was investigated using Matrix Assisted Laser Desorption/Ionization - Time of Flight Mass Spectrometry (MALDI-TOF MS) technique. Saccharomyces cerevisiae, Lactobacillus paracasei, Lactobacillus plantarum, Acetobacter pasteurianus and Acetobacter syzygii were the microbial species identified. S. cerevisiae, L. plantarum, A. pasteurianus and A. syzygii were found in smaller quantities at the beginning of the alcoholic fermentation, but were found throughout the alcoholic and acetic fermentation. Kefir grains were able to utilize apple must as substrate to produce ethanol, and acetic acid. Acetate, volatile alcohols and aldehydes in the vinegar-based kefir were also produced. The yield of acetic acid in the kefir vinegars was ∼79%. The acetic acid concentration was ∼41 g L-1, reaching the required standard for the Brazilian legislation accepts it as vinegar (4.0% acetic acid). Kefir vinegar showed good acceptance in the sensory analysis. The technology proposed here is novel by the application of immobilized-cell biomass (kefir grains) providing a mixed inocula and eliminating the use of centrifuge at the end of the fermentative process. This step will save energy demand and investment. This is the first study to produce apple vinegar using kefir grains.

Bacaba chicha is a beverage prepared by the indigenous Umutina people from the bacaba fruit (Oenocarpus bacaba), a purple berry that is rich in fat and carbohydrates, as well as a source of phenolic compounds. In this study, samples of bacaba chicha beverage were collected, and the microbial community was assessed using culture-dependent and -independent techniques. The nutritional composition and metabolite profiles were analyzed, and species belonging to lactic acid bacteria (LAB) and yeasts were detected. The LAB group detected by culture-dependent analysis included Enterococcus hormaechei and Leuconostoc lactis. Polymerase chain reaction and denaturing gradient gel electrophoresis (PCR-DGGE) detected additional Propionibacterium avidum, Acetobacter spp., and uncultured bacteria. Pichia caribbica and Pichia guilliermondii were detected in a culture-dependent method, and Pichia caribbica was confirmed by PCR-DGGE analysis. The pH value of the beverage was 6.2. The nutritional composition was as follows: 16.47 ± 0.73 g 100 mL-1 dry matter, 2.2 ± 0.0 g 100 mL-1 fat, 3.36 ± 0.44 g 100 mL-1 protein, and 10.87 ± 0.26 g 100 mL-1 carbohydrate. The metabolites detected were 2.69 g L-1 succinic acid, 0.9 g L-1 acetic acid, 0.49 g L-1 citric acid, 0.52 g L-1 ethanol, and 0.4 g L-1 glycerol. This is the first study to identify microbial diversity in bacaba chicha spontaneous fermentation. This study is also the starting step in the immaterial record of this Brazilian indigenous beverage prepared from bacaba fruit.