Potentially probiotic lactic acid bacteria isolated from unpasteurized milk

Isabela Sguilla Rotta, Marcela Fernandes da Matta, Celso Tadeu Barbosa dos Santos, Aline Dias Paiva, Alessandra Barbosa Ferreira Machado

Abstract


Some Lactic Acid Bacteria (LAB), when ingested in adequate quantities, are able to positively impact the host, being considered probiotics. To be used, these bacteria must have their identity known and show specific characteristics. In this study, LAB were isolated from unpasteurized milk samples and characterized in vitro for probiotic potential. After isolation, the colonies were analyzed by Gram stain and catalase test; total DNA was extracted, and the 16S rDNA was amplified and sequenced. Phylogenetic analysis was performed to reconstruct the evolutionary history of the isolates. The isolates were evaluated regarding the resistance to acidic pH and the presence of bile salts. The antagonistic activity of the isolated bacteria was evaluated by the overlay test and the antimicrobial susceptibility profile was performed by the disc diffusion method. Of the 13 isolated bacteria, three were characterized as Gram positive and catalase negative rods, showing 99% of identity with Lactobacillus casei, Lactobacillus paracasei, and Weisella paramesenteroides. The isolates showed in vitro resistance to acidic stress and to the presence of bile salts; showed similar antimicrobial resistance profile and ability to inhibit the growth of potentially pathogenic bacteria, such as Staphylococcus saprophitycus, Escherichia coli, Staphylococcus aureus, Enterococcus faecalis, and Enterococcus faecium. In view of the proven characteristics, the lactic acid bacteria isolated in this work have probiotic potential.


Keywords


phylogenetic analysis; microbiota; Lactobacillus.

References


ALVIM, L. B. Identificação molecular e seleção de bactérias láticas com potencial probiótico isoladas de diferentes mucosas de suínos. 2011. 66 f. Dissertação (Mestrado em Genética) – Universidade Federal de Minas Gerais, Belo Horizonte, 2011.

BOOTH, P. B. et al. Selective depression of blood group antigens associated with hereditary ovalocytosis among Melanesians. Vox sanguinis, v. 32, n. 2, p. 99-110, 1977. DOI: 10.1111/j.1423-0410.1977.tb00612.x

CARVALHO, P. T. et al. Análises de bactérias ácidos láticas, de pH e acidez em amostras de leites fermentados comercializados no município de Sete Lagoas-MG. Brazilian Journal of Food Research, v. 8, n. 3, p.12-21, 2017. DOI: 10.3895/rebrapa.v8n3.3989

CHARTERIS, A. et al. Antibiotic susceptibility of potentially probiotic Lactobacillus Specie. Journal of Food Protection, v. 61, n. 12, p. 1636-1643, 1998. DOI: 10.4315/0362-028x-61.12.1636

CLSI - Clinical and Laboratory Standards Institute. M100-S26: Performance standards for antimicrobial susceptibility testing. 26. ed. Wayne: CLSI, 2017. 256 p.

COPPOLA, R.; TURNES, C. Probióticos e resposta imune. Ciência Rural, v. 34, n. 4, p.1297-1303, 2004. DOI: 10.1590/s0103-84782004000400056

CRUZ, G. B. V. et al. Prospecção tecnológica de micro-organismos probióticos com atividade imunomoduladora. Cadernos de Prospecção, v. 8, n. 4, p. 684-690, 2015. DOI: 10.9771/s.cprosp.2015.008.076

ERKKILÄ, S.; PETÄJÄ, E. Screening of commercial meat starter culture at low pH and in presence of bile salts for potential probiotic use. Meat Science, v. 55, n. 3, p. 297-300, 2000. DOI: 10.1016/S0309-1740(99)00156-4

FAO - Food and Agriculture Organization of the United Nations; WHO - World Health Organization. Health and nutritional properties of probiotics in food including powder milk with live lactic acid bacteria. Report of a Joint FAO/WHO Expert Consultation on evaluation of health and nutritional properties of probiotics in food including powder milk with live lactic acid bacteria. Córdoba: FAO; WHO, 2001. 34 p. Disponível em: http://www.fao.org/tempref/docrep/fao/meeting/009/y6398e.pdf

FERREIRA, C. L. L. F. Grupo de bactérias lácticas e aplicação tecnológica de bactérias probióticas. In: FERREIRA, C. L. L. F. (ed.). Prebióticos e Probióticos: atualização e prospecção. 1. ed. Rio de Janeiro: Editora Rubio, 2012. cap. 1. p. 1-28.

FUSCO, V. et al. The genus Weissella: taxonomy, ecology and biotechnological potential. Frontiers in Microbiology, v. 6, p. 1-22, 2015. DOI: 10.3389/fmicb.2015.00155

GUIMARÃES, F. et al. Seleção de bactérias láticas com potencial probiótico provenientes do leite de transição bovino fermentado. Interciência, v. 43, n. 2, p. 132-136, 2018.

KANT, R.; PALVA, A.; OSSOWSKI, I. An in silico pan-genomic probe for the molecular traits behind Lactobacillus ruminis gut autochthony. Plos One, v. 12, n. 4, p. 1-26, 2017. DOI: 10.1371/journal.pone.0175541

LECLERCQ, S. et al. Low-dose penicillin in early life induces long-term changes in murine gut microbiota, brain cytokines and behavior. Nature Communications, v. 8, n. 15062, p. 1-12, 2017. DOI: 10.1038/ncomms15062

MANGONI, J. et al. Potencial probiotic lactobacilli of pig origin. Acta Scientiarum-Animal Sciences, v. 33, n. 3, p. 267-272, 2011. DOI: 10.4025/actascianimsci.v33i3.9826

MONTORO, D. T. et al. A revised airway epithelial hierarchy includes CFTR-expressing ionocytes. Nature, v. 560, n. 7718, p. 319-324, 2018. DOI: 10.1038/s41586-018-0393-7

OLIVEIRA, G. L. V. et al. Intestinal dysbiosis and probiotic applications in autoimune diseases. Immunology, v. 152, n. 1, p. 1-12, 2017. DOI: 10.1111/imm.12765

OLIVEIRA, S. D. et. al. Detection and identification of Salmonella from poultry-related samples by PCR. Veterinary Microbiology, v. 87, n. 1, p. 25-35, 2002. DOI: 10.1016/s0378-1135(02)00028-7

OUOBA, L. I. I.; LEI, V.; JENSEN, L. B. Resistance of potential probiotic lactic acid bacteria and bifidobacteria of African and European origin to antimicrobials: Determination and transferability of the resistance genes to other bacteria. International Journal of Food Microbiology, v. 121, n. 2, p. 217-224, 2008. DOI: 10.1016/j.ijfoodmicro.2007.11.018

PASSOS, M. C. F.; MORAES-FILHO, J. P. Intestinal microbiota in digestive diseases. Arquivos de Gastroenterologia, v. 54, n. 3, p. 255-262, 2017. DOI: 10.1590/s0004-2803.201700000-31

PENNACCHIA, C. et. al. Selection of Lactobacillus strains from fermented sausages for their potential use as probiotics. Meat Science, v. 67, n. 2, p. 309-317, 2004. DOI: 10.1016/j.meatsci.2003.11.003

PINTO, R. M. S. P. Efeito metabólico e dos probióticos, na microbiota intestinal, na prevenção do cancro do colo-rectal. 2017. 18 f. Trabalho de Conclusão de Curso (Graduação em Nutrição) – Faculdade de Ciência da Nutrição e Alimentação da Universidade do Porto, Porto, 2017.

POSADA, D.; BUCKEY T. R. Model selection and model averaging in phylogenetics: advantages of AKAIKE information criterion and Bayesian approaches over likelihood ratio tests. Systematic Biology, v. 53, n. 5, p. 793-808, 2004. DOI: 10.1080/10635150490522304

RODRIGUEZ-ALONSO, P. et al. Antibiotic resistance in Lactic Acid Bacteria and Micrococcaceae/Staphylococcaceae isolates from artesanal raw milk cheeses, and potential implications on cheese making. Journal of Food Science, v. 74, n. 6, p. 284-293, 2009. DOI: 10.1111/j.1750-3841.2009.01217.x

SANSCHAGRIN, S.; YERGEAU, E. Next-generation sequencing of 16S ribosomal RNA gene amplicons. Journal of Visualized Experiments, v. 90, n. 51709, p. 1-6, 2014. DOI: 10.3791/51709

SILVA, J. G. Identificação molecular de bactérias ácido láticas e propriedades probióticas in vitro de Lactobacillus spp. isolados de queijo Minas artesanal de Araxá, Minas Gerais. 2016. 82 f. Dissertação (Mestrado em Tecnologia e Inspeção de Produtos de Origem Animal) – Escola de Veterinária da Universidade Federal de Minas Gerais, Belo Horizonte, 2016.

SUÁREZ, J. E. Microbiota autóctona, probióticos y prebióticos. Nutrición Hospitalaria, v. 31, n. supl. 1, p. 3-9, 2015. DOI: 10.3305/nh.2015.31.sup1.8701

SUZUKI, M.; RAPPÉ, M. S.; GIOVANNONI, S. J. Kinetic bias in estimates of coastal picoplankton community structure obtained by measurements of small-subunit rRNA gene PCR amplicon length heterogeneity. Applied and Environmental Microbiology, v. 64, n. 11, p. 4522-4529, 1998.

VENTURA, M.; TURRONI, F.; VAN SINDEREN, D. Probiogenomics as a tool to obtain genetic insights into adaptation of probiotic bacteria to the human gut. Bioengineered Bugs, v. 3, n. 2, p. 73-79, 2012. DOI: 10.4161/bbug.18540

WENDLING, L. K.; WESCHENFELDER, S. Probióticos e alimentos lácteos fermentados – uma revisão. Revista do Instituto de Laticínios Cândido Tostes, v. 68, n. 395, p. 49-57, 2013. DOI: 10.5935/2238-6416.20130048




DOI: https://doi.org/10.14295/2238-6416.v75i3.820

Refbacks

  • There are currently no refbacks.


Licença Creative Commons
Esta obra está licenciada com uma Licença Creative Commons Atribuição 4.0 Internacional.