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Isolation and characterization of Lactobacillus spp. from oral cavity and its probiotic action in Candida albicans and Streptococcus mutans

Grant number: 15/09770-9
Support type:Regular Research Grants
Duration: November 01, 2015 - October 31, 2017
Field of knowledge:Health Sciences - Dentistry
Principal Investigator:Juliana Campos Junqueira
Grantee:Juliana Campos Junqueira
Home Institution: Instituto de Ciência e Tecnologia (ICT). Universidade Estadual Paulista (UNESP). Campus de São José dos Campos. São José dos Campos , SP, Brazil

Abstract

The objective of this study will be isolate and identify strains of Lactobacillus spp. from the oral cavity of individuals caries free and to evaluate their probiotic action in Candida albicans and Streptococcus mutans by in vitro and in vivo studies. Saliva samples will be collected and identified by the API50CHL system and confirmed by Polymerase Chain Reaction (PCR). Thirty strains identified as Lactobacillus will be submitted to the tests for the study of inhibitory action on C. albicans and S. mutans. For the C. albicans study, it will be evaluated the in vitro effects of Lactobacillus strains on biofilm formation and on the ability of C. albicans filamentation. Next, ten strains that having better inhibitory action on C. albicans will be selected for the in vivo study using host models of G. mellonella and C. elegans. In the G. mellonella model, the experimental infection will be analyzed by survival curves, quantification of fungal load in the hemolymph, and hemocyte density quantification. For the C. elegans model, the infection will be evaluated by survival curves, count of CFU/mL in the intestinal tract of nematodes and study of C. albicans filamentation. In addition, it will be performed the quantification of gene expression of EFG1, ALS1 and HWP1 in heterotypic and monotypic biofilms of C. albicans by testing Real Time PCR (qPCR). In relation to S. mutans study, the thirty Lactobacillus strains will be submitted to tests of antibacterial activity against S. mutans in planktonic growth. Hereafter, 10 strains that exhibit higher antibacterial activity will be selected for the study of antimicrobial effects on biofilms of S. mutans formed on hydroxyapatite discs. For in vivo study, one Lactobacillus strain with higher inhibitory effect on biofilm of S. mutans will be selected to assess its effects on the development of experimental caries in rats. After infection by S. mutans and Lactobacillus strain in the oral cavity of the rats, sampling of biofilm will be collected to count CFU/mL and analysis of the maxilla and mandible in stereomicroscope for quantification of occlusal caries and smooth surface will be performed. The results will be submitted to an exploratory analysis for selecting the most appropriate statistical test for each experiment of this study. (AU)

Scientific publications (8)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
SANTOS, RAFAELLA BRAGA; SCORZONI, LILIANA; NAMBA, ANDRESSA MAYUMI; ROSSONI, RODNEI DENNIS; CARDOSO JORGE, ANTONIO OLAVO; JUNQUEIRA, JULIANA CAMPOS. Lactobacillus species increase the survival of Galleria mellonella infected with Candida albicans and non-albicans Candida clinical isolates. Medical Mycology, v. 57, n. 3, p. 391-394, APR 2019. Web of Science Citations: 4.
ROSSONI, RODNEI DENNIS; VELLOSO, MARISOL DOS SANTOS; DE BARROS, PATRICIA PIMENTEL; DE ALVARENGA, JANAINA ARAUJO; DOS SANTOS, JESSICA DIANE; CHIPOLETTI DOS SANTOS PRADO, ANA CAROLINA; RIBEIRO, FELIPE DE CAMARGO; ANBINDER, ANA LIA; JUNQUEIRA, JULIANA CAMPOS. Inhibitory effect of probiotic Lactobacillus supernatants from the oral cavity on Streptococcus mutans biofilms. Microbial Pathogenesis, v. 123, p. 361-367, OCT 2018. Web of Science Citations: 3.
ROSSONI, RODNEI DENNIS; VELLOSO, MARISOL DOS SANTOS; ALVES FIGUEIREDO, LIVIA MARA; MARTINS, CAROLINA PISTILLE; CARDOSO JORGE, ANTONIO OLAVO; JUNQUEIRA, JULIANA CAMPOS. Clinical strains of Lactobacillus reduce the filamentation of Candida albicans and protect Galleria mellonella against experimental candidiasis. Folia Microbiologica, v. 63, n. 3, p. 307-314, MAY 2018. Web of Science Citations: 7.
DE BARROS, PATRICIA PIMENTEL; SCORZONI, LILIANA; RIBEIRO, FELIPE DE CAMARGO; DE OLIVEIRA FUGISAKI, LUCIANA RUANO; FUCHS, BETH BURGWYN; MYLONAKIS, ELEFTHERIOS; CARDOSO JORGE, ANTONIO OLAVO; JUNQUEIRA, JULIANA CAMPOS; ROSSONI, RODNEI DENNIS. Lactobacillus paracasei 28.4 reduces in vitro hyphae formation of Candida albicans and prevents the filamentation in an experimental model of Caenorhabditis elegans. Microbial Pathogenesis, v. 117, p. 80-87, APR 2018. Web of Science Citations: 7.
DE BARROS, PATRICIA PIMENTEL; ROSSONI, RODNEI DENNIS; FREIRE, FERNANDA; RIBEIRO, FELIPE DE CAMARGO; DAS CHAGAS LOPES, LUCAS ALEXANDRE; JUNQUEIRA, JULIANA CAMPOS; CARDOSO JORGE, ANTONIO OLAVO. Candida tropicalis affects the virulence profile of Candida albicans: an in vitro and in vivo study. PATHOGENS AND DISEASE, v. 76, n. 2 MAR 2018. Web of Science Citations: 5.
JORJAO, ADELINE L.; OLIVEIRA, LUCIANE D.; SCORZONI, LILIANA; FIGUEIREDO-GODOI, LIVIA MARA A.; PRATA, MARCIA CRISTINA A.; JORGE, ANTONIO OLAVO C.; JUNQUEIRA, JULIANA C. From moths to caterpillars: Ideal conditions for Galleria mellonella rearing for in vivo microbiological studies. VIRULENCE, v. 9, n. 1, p. 383-389, 2018. Web of Science Citations: 12.
ROSSONI, RODNEI DENNIS; DE BARROS, PATRICIA PIMENTEL; DE ALVARENGA, JANAINA ARAUJO; RIBEIRO, FELIPE DE CAMARGO; VELLOSO, MARISOL DOS SANTOS; FUCHS, BETH BURGWYN; MYLONAKIS, ELEFTHERIOS; CARDOSO JORGE, ANTONIO OLAVO; JUNQUEIRA, JULIANA CAMPOS. Antifungal activity of clinical Lactobacillus strains against Candida albicans biofilms: identification of potential probiotic candidates to prevent oral candidiasis. BIOFOULING, v. 34, n. 2, p. 212-225, 2018. Web of Science Citations: 17.
ROSSONI, RODNEI DENNIS; FUCHS, BETH BURGWYN; DE BARROS, PATRICIA PIMENTEL; VELLOSO, MARISOL DOS SANTOS; CARDOSO JORGE, ANTONIO OLAVO; JUNQUEIRA, JULIANA CAMPOS; MYLONAKIS, ELEFTHERIOS. Lactobacillus paracasei modulates the immune system of Galleria mellonella and protects against Candida albicans infection. PLoS One, v. 12, n. 3 MAR 7 2017. Web of Science Citations: 22.

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