Atividade antimicrobiana de kefir fermentado com subproduto de uva contra Alicyclobacillus acidoterrestris / Antimicrobial activity of fermented kefir with a byproduction

Jéssica Lima de Menezes, Amanda Gouveia Mizuta, Tatiane Viana Dutra, Taiana Varela Ferreira, Edinéia Bonin, Vicky Cristine Bragante Thumaz, Márcia Maria dos Anjos Szczerepa, Benício Alves de Abreu Filho


Alicyclobacillus spp. they are spore-forming bacteria that deteriorate acidic fruit-based drinks, causing economic losses, with A. acidoterrestris being the most studied, responsible for causing sensory changes, especially in orange juice. The use of natural antimicrobial agents in foods can be an option with great advantages for the industry and the consumer. Therefore, the objective of this work was to investigate the antimicrobial activity of extracts fermented by kefir, from a grape by-product, against A. acidoterrestris. Four extracts were prepared with kefir grains, being 1 - grape extract and kefir grains; 2 - grape extract, brown sugar and kefir grains; 3 - grape extract, ultrasound and kefir grains; and 4 - grape extract, ultrasound, brown sugar and kefir grains. The four extracts were fermented at 28 ºC for 7 days. The extracts were centrifuged at 10,000 rpm for 10 min and the supernatant was subjected to membrane filtration (0.22 μm). The minimum inhibitory concentration (MIC) was determined by the serial microdilution technique of extracts of 50 to 0.1% concentration in Bacillus acidoterrestris (BAT) medium. The minimum bactericidal concentration (CBM) was also determined. Structural changes in cells after treatment were evaluated by scanning electron microscopy (SEM). The MIC value capable of inhibiting the visible growth of A. acidoterrestris for all extracts was 1.6%, while the CBM was 50% for extracts 1 and 3, while for extracts 2 and 4 the CBM was 25%. The results show that extracts 2 and 4 obtained better CBM value, possibly because kefir produced more secondary metabolites with the addition of brown sugar, in addition, the use of ultrasound did not interfere. The results of the inhibitory and/or bactericidal concentration indicate that the extracts have activity against A. acidotrrestris.


deterioration, preservation, scanning electron microscopy.

Full Text:



ANJOS, M.M., SILVA, A.A., PASCOLI, I.C., MIKCHA, J.M.G., MACHINSKI JR, M.; PERALTA, R.M., FILHO, B.A.A. Antibacterial activity of papain and bromelain on Alicyclobacillus spp. International Journal of Food Microbiology, v. 216, p. 121–126, 2016.

CLSI - CLINICAL AND LABORATORY STANDARDS INSTITUTE. Methods for Diluition Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically; Approved Standard. 11th edition. (CLSI document M07- A11), 2018.

COMA, M. E.; PELTZER, M. A.; DELGADO, J. F.; SALVAY, A. G. Water kefir grains as an innovative source of materials: Study of plasticiser content on film properties. European Polymer Journal, v. 120, 2019.

DEINHARD, G.; BLANZ, P.; PORALLA, K.; ALTAN, E. Bacillus acidoterrestris sp. nov., a new thermotolerant acidophile isolated from different soils. Systematic and Applied Microbiology, v. 10, p. 47-53, 1987.

DUTRA, T. V.; CASTRO, J. C.; MENEZES, J. L.; RAMOS, T. R.; PRADO, I. N.; MACHINSKI JR, M.; MIKCHA, J. M. G.; ABREU FILHO, B. A. Bioactivity of oregano (Origanum vulgare) essential oil against Alicyclobacillus spp. Industrial Crops and Products, v. 129, p. 345-349, 2019.

ENDO, H.E., CORTEZ, D.A.G., UEDA-NAKAMURA, T., NAKAMURA, C.V., DIAS FILHO, B.P. Potent antifungal activity of extracts and pure compound isolated from pomegranate peels and synergism with fluconazole against Candida albicans. Research in. Microbiology, v. 161, p. 534–540, 2010.

FIORDA, F. A.; DE MELO PEREIRA, G. V.; THOMAZ-SOCCOL, V.; MEDEIROS, A. P.; RAKSHIT, S. K.; SOCCOL, C. R. Development of kefir-based probiotic beverages with DNA protection and antioxidant activities using soybean hydrolyzed extract, colostrum and honey. LWT - Food Science and Technology, v. 86, p. 690–607, 2016.

GAWARE, V.; KOTADE,K.; DOLAS, R.; DHAMAK, K.; SOMWANSHI, S.; NIKAM, V.; KHADSE, A.; KASHID, V. The magic of kefir: a review. Pharmacology, v. 1, p. 376-386, 2011.

GOLOWCZYC, M. A.; MOBILI, P.; GARROTE, G. L.; ABRAHAM, A. G.; ANTONI, G. L. Protective action of Lactobacillus kefir carrying S-layer protein against Salmonella enterica serovar Enteritidis. International Journal of Food Microbiology, v. 118, p. 264-273, 2007.

HU, X.; HUANG, E.; BARRINGER, A. S.; YOUSEF, E. A. Factors affecting Alicyclobacillus acidoterrestris growth and guaiacol production and controlling apple juice spoilage by lauric arginate and ϵ-polylysine. LWT - Food Science and Technology, v. 119, 2020.

HUERTAS, J. P.; ESTEBAN, M. D.; ANTOLINOS, V.; PALOP, A. Combined effect of natural antimicrobials and thermal treatments on Alicyclobacillus acidoterrestris spores. Food Control, v. 35, n. 1, p. 73-78, 2014.

JARDIM, M. F. A.; FURLAN, L. C. O.; CARVALHO, I. S.; BARBOSA, L. N.; OTUTUMI, L. K.; JACOMASSI, E. GERMANO, R. M. SOARES. A. A. Antibacterial and antioxidant activity of water extracts of zingiber officinale roscoe leaves and rhizomas cultivated in unipar's medical court. Brazilian Journal of Development, v. 5, n. 10, p. 18292-18309, 2019.

LIMA, I. A. S. I.; RIBEIRO, I. A.; NERIS, T. S.; SILVA, S. S.; LOSS, R. A.; GUEDES, S. F. Extração de compostos fenólicos do noni (Morinda citrifolia Linn) empregando energia ultrassônica. Scientia Plena, v. 14, 2018.

MIAO, J.; LIU, G.; KE, C.; FAN, W.; LI, C.; CHEN, Y.; DIXON, W.; SONG, M.; CAO Y.; XIAO, H. Inhibitory effects of a novel antimicrobial peptide from kefir against Escherichia coli. Food Control, v. 65, p. 63-72, 2016.

MOLVA, C.; BAYSAL, A. H. Antimicrobial activity of grape seed extract on Alicyclobacillus acidoterrestris DSM 3922 vegetative cells and spores in apple juice. LWT - Food Science and Technology, v. 60, n. 1, p. 238-245, 2015.

PASCOLI, I. C.; ANJOS, M. M.; SILVA, A. A.; LORENZETTI, F. B.; CORTEZ, D. A. G.; MIKCHA, J. M. G.; NAKAMURA T. U.; NAKAMURA, C. V.; ABREU FILHO B. A. Piperaceae extracts for controlling Alicyclobacillus acidoterrestris growth in commercial orange juice. Industrial Crops and Products, v. 116, p. 224-230, 2018.

PISKERNIK, S.; KLANČNIK, A.; DEMŠAR, L.; MOŽINA, S. S.; JERŠEK B. Control of Alicyclobacillus spp. vegetative cells and spores in apple juice with rosemary extracts. Food Control, v. 60, p. 205-214, 2016.

RODRIGUES K. L.; CAPUTO L. R. G.; CARVALHO J. C. T.; EVANGELISTA J.; SCHNEEDORF, J. M. Antimicrobial and healing activity of kefir and kefiran extract. International Journal of Antimicrobial Agents, v. 25, n. 5, 2005, p. 404-408, 2005.

SAVASTANO, M. L.; PATI S.; BEVILACQUA, A.; CORBO M. R.; RIZZUTI, A.; PISCHETSRIEDER, M.; LOSITO, I. Influence of the production technology on kefir characteristics: Evaluation of microbiological aspects and profiling of phosphopeptides by LC-ESI-QTOF-MS/MS. Food Research International, v. 129, 2020.

ZAMBRANO C.; KEREKES E. B.; KOTOGÁN A.; PAPP T.; VÁGVÖLGYI C.; KRISCH J.; TAKÓ M. Antimicrobial activity of grape, apple and pitahaya residue extracts after carbohydrase treatment against food-related bacteria. LWT - Food Science and Technology, v. 100, p. 416-425, 2019.



  • There are currently no refbacks.