Isolation and characterization of Lactobacillus spp. from oral cavity and their probiotic action in Candida albicans: biofilm formation, infection in invertebrate models and EFG1, HWP1 and ALS1 gene expression

• Grant number: 13/25181-8
• Support Opportunities: Scholarships in Brazil - Doctorate
• Effective date (Start): March 01, 2014
• Effective date (End): December 31, 2017
• Field of knowledge: Health Sciences - Dentistry
• Principal Investigator: Juliana Campos Junqueira
• Grantee: Rodnei Dennis Rossoni
• Host Institution: Instituto de Ciência e Tecnologia (ICT). Universidade Estadual Paulista (UNESP). Campus de São José dos Campos. São José dos Campos , SP, Brazil
• Associated scholarship(s): 14/12458-4 - Probiotic action of Lactobacillus spp. on Candida albicans infection: study in invertebrate model hosts, BE.EP.DR

Abstract

The objective of this study will be to isolate and identify strains of Lactobacillus spp. from oral cavity of individuals caries free and evaluate their probiotic action in Candida albicans by studies in vitro and in vivo. Saliva samples will be collected and identified by the API50CHL system. Firstly, the effects of all strains identified as Lactobacillus on biofilm formation and on the ability of filamentation of C. albicans in vitro will be evaluated. Mixed biofilms will be formed on the bottom of 96-well plates for 48 hours. Then, the amount of biofilm will be analyzed by counting the colony forming units (CFU/mL). Filamentation assays will be performed on 24-well plates and analysed by optical microscopy. Next, ten strains that having better inhibitory action on C. albicans will be selected for the execution of the other tests. For in vivo study, Lactobacillus strains will be inoculated with C. albicans in Galleria mellonella caterpillars and the experimental infection will be evaluated by survival curves, quantification of fungal load in the hemolymph, and hemocyte density quantification. For the Caenorhabditis elegans model, the nematodes will be co-infected with C. albicans and Lactobacillus. After that, the infection will be evaluated by survival curves, count of CFU/mL in the intestinal tract of nematodes and study of filamentation. The quantification of gene expression of target genes EFG1, ALS1 and HWP1 will be performed after 48 h in heterotypic and monotypic biofilms of C. albicans by testing Real Time PCR (qPCR). The results will be analyzed for selecting the best statistical test for each analysis of this study, considering a significance level of 5 % test. (AU)

Scientific publications

(7)

(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)

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. (14/11841-9, 13/25181-8, 17/19219-3, 15/09770-9)

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. (13/26820-4, 15/09770-9, 13/25181-8)

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. (15/09770-9, 13/25181-8)

RODNEI DENNIS ROSSONI; PATRÍCIA PIMENTEL DE BARROS; FERNANDA FREIRE; JÉSSICA DIANE DOS SANTOS; ANTONIO OLAVO CARDOSO JORGE; JULIANA CAMPOS JUNQUEIRA. Study of Microbial Interaction Formed by "Candida krusei" and "Candida glabrata": "In Vitro" and "In Vivo" Studies. Brazilian Dental Journal, v. 28, n. 6, p. 669-674, 2017-12-00. (12/02184-9, 13/25181-8)

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. (14/12458-4, 13/25181-8, 15/09770-9)

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. (13/25181-8, 15/09770-9, 12/15250-0)

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. (15/09770-9, 13/25181-8, 14/12458-4)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

ROSSONI, Rodnei Dennis. Isolation and characterization of Lactobacillus spp. from oral cavity and their probiotic action in Candida albicans: biofilm formation, infection in invertebrate models and EFG1, HWP1 and ALS1 gene expression. 2017. Doctoral Thesis - Universidade Estadual Paulista (Unesp). Instituto de Ciência e Tecnologia. São José dos Campos São José dos Campos.