Microbiology, Applied Microbiology and Biotechnology, Molecular Biology, Microbiology (medical)
40
Scopus Publications
Scopus Publications
Development of Melting-Curve-Based Real-Time PCR for Differentiating Medically Important Candida Species Eliandro Reis Tavares, Virginia Prezzi Santos, Isabela Madeira de Castro, Paulo Henrique Guilherme Borges, Gislaine Silva-Rodrigues, Anna Paula Silva Olak, Guilherme Bartolomeu-Gonçalves, Jussevania Pereira Santos, Alexandre Tadachi Morey, Kelly Ishida, Sueli Fumie Yamada-Ogatta, Lucy Megumi Yamauchi International Journal of Molecular Sciences, 2025 Candida species are the primary fungal pathogens of invasive infections associated with high morbidity and mortality. The identification of these microorganisms is critical for therapeutic management and control of hospital infection. Herein, assays targeting the Intergenic Spacer 2 (IGS2) and Internal Transcribed Spacer 1 (ITS1) from the rDNA locus were developed to differentiate Candida species. Based on consensus nucleotide sequences, specific primers and positive controls were designed, and standard PCR and real-time PCR (qPCR) assays were performed. All primers resulted in specific amplification of the molecular targets of each species with no amplifications of the negative template control. Furthermore, the primers were highly specific when tested with a range of fungal DNAs and no cross-reactivity was observed among Candida species. The assays presented a limit of detection (LoD) of 10 copies of positive control per reaction for all specific primers designed. Overall, our results showed that qPCR assays employing primers targeting the regions IGS2 and ITS1 completely differentiated between Candida albicans, Candida auris, Candida parapsilosis, Candida tropicalis, and Nakazeomyces glabratus, with great accuracy and no amplification of DNA from other fungal species.
Synergistic Antibacterial Effect of Eugenol and Biogenic Silver Nanoparticles on Staphylococcus pseudintermedius Isolated from Canine Keratoconjunctivitis Sicca Weslei Roberto Correia Cabral, Caio Ferreira de Oliveira, Rogerio Giuffrida, Lais Fernanda de Almeida Spoladori, Isabela Madeira de Castro, Guilherme Bartolomeu-Gonçalves, Helena Tiemi Suzukawa, Gabriella Maria Andriani, Gerson Nakazato, Eliandro Reis Tavares, Lucy Megumi Yamauchi, Sueli Fumie Yamada-Ogatta Molecules, 2025 Plants are a valuable source of bioactive compounds with therapeutic potential. Antibacterials of natural origin represent a promising and sustainable alternative in the fight against bacterial infections. In addition to being effective against bacterial growth, these natural agents may have lower toxicity and fewer side effects, which reinforces their value in the development of new therapeutic strategies. This study reports on the antibacterial effect of eugenol (EUG) and biogenic silver nanoparticles (bioAgNPs) synthesized using the aqueous extract of Trichilia catigua A. Juss. bark, alone or in combination, against planktonic and sessile cells of multidrug-resistant Staphylococcus pseudintermedius, one of the main opportunistic pathogens in dogs. EUG and bioAgNPs showed a dose- and time-dependent bactericidal effect on planktonic cells, interfering with cell membrane integrity. The interaction between EUG and bioAgNPs was classified as synergistic or indifferent for planktonic cells. Except for one isolate, the combination exhibited a synergistic effect for biofilms previously formed on abiotic surfaces for 24 h. Both bioactive compounds promoted morphological and ultrastructural changes in S. pseudintermedius biofilms. All concentrations of EUG and bioAgNPs in synergistic or indifferent combinations showed reduced toxicity to mammalian cells. These findings suggest that the EUG and bioAgNP combination could be a promising strategy for controlling S. pseudintermedius infections.
Strategy for the Construction of SARS-CoV-2 S and N Recombinant Proteins and Their Immunogenicity Evaluation Paulo Henrique Guilherme Borges, Barbara Gregio, Helena Tiemi Suzukawa, Gislaine Silva-Rodrigues, Emanuella de Castro Andreassa, Isabela Madeira de Castro, Guilherme Bartolomeu-Gonçalves, Emerson José Venancio, Phileno Pinge-Filho, Viviane Monteiro Góes, Celso Vataru Nakamura, Eliandro Reis Tavares, Tatiana de Arruda Campos Brasil de Souza, Sueli Fumie Yamada-Ogatta, Lucy Megumi Yamauchi Biotech, 2025 This study reports the construction, expression, and purification of synthetic SARS-CoV-2 spike (S) and nucleoprotein (N) containing immunodominant epitopes. The pET28aS_epit construct included epitopes 287–317, 402, 507, 524–598, and 601–640, while the pET28aN_epit construct included residues 42–62, 153–172, and 355–401. Commercial sequences of both proteins were used as controls. The four constructs were expressed using the Escherichia coli BL21(DE3) star strain at 37 °C. The results show that the S protein constructs were insoluble, unlike the N protein constructs. Both recombinant proteins induced immune responses in mice and were recognized by antibodies present in sera from COVID-19-positive and/or SARS-CoV-2-vaccinated humans. No significant differences in immune recognition were observed between our constructs and the commercially available proteins. In conclusion, S_epit and N_epit could be promising starting points for the development of new strategies based on immunological reactions for the control of SARS-CoV-2 infections.
Synergistic Antibacterial Interaction of Geraniol and Biogenic Silver Nanoparticles on Methicillin-Resistant Staphylococcus aureus Isabela Madeira de Castro, Camila Antunes, Camila Cristina Valentim, Laís Fernanda de Almeida Spoladori, Helena Tiemi Suzukawa, Guilherme Ferreira Correia, Gislaine Silva-Rodrigues, Paulo Henrique Guilherme Borges, Guilherme Bartolomeu-Gonçalves, Mariana Luiza Silva, Marcelle de Lima Ferreira Bispo, Rayanne Regina Beltrame Machado, Celso Vataru Nakamura, Gerson Nakazato, Phileno Pinge-Filho, Eliandro Reis Tavares, Lucy Megumi Yamauchi, Sueli Fumie Yamada-Ogatta Plants, 2025 Since ancient times, plants have been used in folk medicine to treat different diseases. Plants offer exceptional chemical diversity with a wide range of biological activities, and have therefore been the most promising sources for the discovery and development of drugs, including antimicrobial agents. This study reports the antibacterial effect of geraniol (GER), alone and in combination with biogenic silver nanoparticles (bioAgNPs), produced using the aqueous extract of Trichilia catigua bark, against planktonic and sessile cells of methicillin-resistant Staphylococcus aureus (MRSA), one of the main opportunistic and potentially fatal human pathogens. GER had a time-dependent bactericidal effect on planktonic cells, impairing the cell membrane integrity. In addition, GER inhibited the staphyloxanthin production, and molecular docking analyses supported the in silico affinity of GER to dehydrosqualene synthase (CrtM) and 4,4′-diaponeurosporen-aldehyde dehydrogenase (AldH), which are key enzymes within the pigment biosynthesis pathway in S. aureus. GER treatment increased the sensitivity of MRSA to hydrogen peroxide killing. GER displayed synergism with bioAgNPs against planktonic and sessile cells, inhibiting bacterial adhesion and the viability of biofilms formed on abiotic surfaces. MRSA planktonic and sessile cells treated with GER or GER/bioAgNPs displayed severe morphological and ultrastructural alterations. Notably, neither GER nor its combination caused in vitro and in vivo toxicity in mammalian cells and Galleria mellonella larvae, respectively. These findings suggest that the combination of GER/bioAgNPs may be a promising strategy to control MRSA infections.
Tuberculosis Diagnosis: Current, Ongoing, and Future Approaches Guilherme Bartolomeu-Gonçalves, Joyce Marinho de Souza, Bruna Terci Fernandes, Laís Fernanda Almeida Spoladori, Guilherme Ferreira Correia, Isabela Madeira de Castro, Paulo Henrique Guilherme Borges, Gislaine Silva-Rodrigues, Eliandro Reis Tavares, Lucy Megumi Yamauchi, Marsileni Pelisson, Marcia Regina Eches Perugini, Sueli Fumie Yamada-Ogatta Diseases, 2024 Tuberculosis (TB) remains an impactful infectious disease, leading to millions of deaths every year. Mycobacterium tuberculosis causes the formation of granulomas, which will determine, through the host–pathogen relationship, if the infection will remain latent or evolve into active disease. Early TB diagnosis is life-saving, especially among immunocompromised individuals, and leads to proper treatment, preventing transmission. This review addresses different approaches to diagnosing TB, from traditional methods such as sputum smear microscopy to more advanced molecular techniques. Integrating these techniques, such as polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP), has significantly improved the sensitivity and specificity of M. tuberculosis identification. Additionally, exploring novel biomarkers and applying artificial intelligence in radiological imaging contribute to more accurate and rapid diagnosis. Furthermore, we discuss the challenges of existing diagnostic methods, including limitations in resource-limited settings and the emergence of drug-resistant strains. While the primary focus of this review is on TB diagnosis, we also briefly explore the challenges and strategies for diagnosing non-tuberculous mycobacteria (NTM). In conclusion, this review provides an overview of the current landscape of TB diagnostics, emphasizing the need for ongoing research and innovation. As the field evolves, it is crucial to ensure that these advancements are accessible and applicable in diverse healthcare settings to effectively combat tuberculosis worldwide.
Geraniol Potentiates the Effect of Fluconazole against Planktonic and Sessile Cells of Azole-Resistant Candida tropicalis: In Vitro and In Vivo Analyses Gislaine Silva-Rodrigues, Isabela Madeira de Castro, Paulo Henrique Guilherme Borges, Helena Tiemi Suzukawa, Joyce Marinho de Souza, Guilherme Bartolomeu-Gonçalves, Marsileni Pelisson, Cássio Ilan Soares Medeiros, Marcelle de Lima Ferreira Bispo, Ricardo Sérgio Couto de Almeida, Kelly Ishida, Eliandro Reis Tavares, Lucy Megumi Yamauchi, Sueli Fumie Yamada-Ogatta Pharmaceutics, 2024 Candida tropicalis is regarded as an opportunistic pathogen, causing diseases ranging from superficial infections to life-threatening disseminated infections. The ability of this yeast to form biofilms and develop resistance to antifungals represents a significant therapeutic challenge. Herein, the effect of geraniol (GER), alone and combined with fluconazole (FLZ), was evaluated in the planktonic and sessile cells of azole-resistant C. tropicalis. GER showed a time-dependent fungicidal effect on the planktonic cells, impairing the cell membrane integrity. Additionally, GER inhibited the rhodamine 6G efflux, and the molecular docking analyzes supported the binding affinity of GER to the C. tropicalis Cdr1 protein. GER exhibited a synergism with FLZ against the planktonic and sessile cells, inhibiting the adhesion of the yeast cells and the viability of the 48-h biofilms formed on abiotic surfaces. C. tropicalis biofilms treated with GER, alone or combined with FLZ, displayed morphological and ultrastructural alterations, including a decrease in the stacking layers and the presence of wilted cells. Moreover, neither GER alone nor combined with FLZ caused toxicity, and both treatments prolonged the survival of the Galleria mellonella larvae infected with azole-resistant C. tropicalis. These findings indicate that the combination of GER and FLZ may be a promising strategy to control azole-resistant C. tropicalis infections.
Development of a Melting-Curve-Based Multiplex Real-Time PCR Assay for the Simultaneous Detection of Viruses Causing Respiratory Infection Eliandro Reis Tavares, Thiago Ferreira de Lima, Guilherme Bartolomeu-Gonçalves, Isabela Madeira de Castro, Daniel Gaiotto de Lima, Paulo Henrique Guilherme Borges, Gerson Nakazato, Renata Katsuko Takayama Kobayashi, Emerson José Venancio, César Ricardo Teixeira Tarley, Elaine Regina Delicato de Almeida, Marsileni Pelisson, Eliana Carolina Vespero, Andrea Name Colado Simão, Márcia Regina Eches Perugini, Gilselena Kerbauy, Marco Aurélio Fornazieri, Maria Cristina Bronharo Tognim, Viviane Monteiro Góes, Tatiana de Arruda Campos Brasil de Souza, Danielle Bruna Leal Oliveira, Edison Luiz Durigon, Lígia Carla Faccin-Galhardi, Lucy Megumi Yamauchi, Sueli Fumie Yamada-Ogatta Microorganisms, 2023 The prompt and accurate identification of the etiological agents of viral respiratory infections is a critical measure in mitigating outbreaks. In this study, we developed and clinically evaluated a novel melting-curve-based multiplex real-time PCR (M-m-qPCR) assay targeting the RNA-dependent RNA polymerase (RdRp) and nucleocapsid phosphoprotein N of SARS-CoV-2, the Matrix protein 2 of the Influenza A virus, the RdRp domain of the L protein from the Human Respiratory Syncytial Virus, and the polyprotein from Rhinovirus B genes. The analytical performance of the M-m-qPCR underwent assessment using in silico analysis and a panel of reference and clinical strains, encompassing viral, bacterial, and fungal pathogens, exhibiting 100% specificity. Moreover, the assay showed a detection limit of 10 copies per reaction for all targeted pathogens using the positive controls. To validate its applicability, the assay was further tested in simulated nasal fluid spiked with the viruses mentioned above, followed by validation on nasopharyngeal swabs collected from 811 individuals. Among them, 13.4% (109/811) tested positive for SARS-CoV-2, and 1.1% (9/811) tested positive for Influenza A. Notably, these results showed 100% concordance with those obtained using a commercial kit. Therefore, the M-m-qPCR exhibits great potential for the routine screening of these respiratory viral pathogens.
Whole Genome Sequencing of an Extensively Drug-Resistant Raoultella planticola Isolate Containing blaKPC-2, blaNDM-1, and blaCTX-M-15 Paulo Victor Batista Marini, Eliandro Reis Tavares, Cintia Werner Motter, Letícia Busato Migliorini, Romário Oliveira de Sales, Nayara Helisandra Fedrigo, Danielle Rosani Shinohara, Mariangela Hungria, Sueli Fumie Yamada-Ogatta, Maria Cristina Bronharo Tognim Microbial Drug Resistance, 2023 Raoultella planticola harboring genes that confer resistance to antimicrobials, such as carbapenems, have been associated with severe infections in immunocompromised patients. In this study, we reported the first whole genome sequence of a Brazilian isolate of R. planticola and the genomic context of antibiotic resistance markers. By whole-genome sequencing (WGS) of a carbapenem-resistant R. planticola isolate, RpHUM1, we found 23 resistance-encoding genes belonging to 9 classes of antibiotics (aminoglycosides, β-lactams, fluoroquinolones, fosfomycin, macrolides, phenicols, sulfonamides, tetracycline, and diaminopyrimidine derivatives) and 3 plasmids (RpHUM1pEaer-4382s, RpHUM1_pFDAARGOS_440, and RpHUM1pRSF1010). This isolate coharbored the genes bla KPC-2 , which is carried by the plasmid RpHUM1pEaer-4382s, and bla NDM-1 and bla CTX-M-15 all located in the accessory genome. In addition, these genes were associated with, at least, one mobile genetic element. This comprehensive knowledge is of great importance for implementation of control measures to prevent the rapid dissemination of this neglected microorganism and their genetic resistance background.
Antiprotozoal Activity of Benzoylthiourea Derivatives against Trypanosoma cruzi: Insights into Mechanism of Action Patrícia Morais Lopes Pereira, Bruna Terci Fernandes, Vitória Ribeiro dos Santos, Weslei Roberto Correia Cabral, Maria Isabel Lovo-Martins, Lais Alonso, César Armando Contreras Lancheros, Jéssica Carreira de Paula, Priscila Goes Camargo, Helena Tiemi Suzukawa, Antônio Alonso, Fernando Macedo, Celso Vataru Nakamura, Eliandro Reis Tavares, Marcelle de Lima Ferreira Bispo, Lucy Megumi Yamauchi, Phileno Pinge-Filho, Sueli Fumie Yamada-Ogatta Pathogens, 2023 For decades, only two nitroheterocyclic drugs have been used as therapeutic agents for Chagas disease. However, these drugs present limited effectiveness during the chronic phase, possess unfavorable pharmacokinetic properties, and induce severe adverse effects, resulting in low treatment adherence. A previous study reported that N-(cyclohexylcarbamothioyl) benzamide (BTU-1), N-(tert-butylcarbamothioyl) benzamide (BTU-2), and (4-bromo-N-(3-nitrophenyl) carbamothioyl benzamide (BTU-3) present selective antiprotozoal activity against all developmental forms of Trypanosoma cruzi Y strain. In this study, we investigated the mechanism of action of these compounds through microscopy and biochemical analyses. Transmission electron microscopy analysis showed nuclear disorganization, changes in the plasma membrane with the appearance of blebs and extracellular arrangements, intense vacuolization, mitochondrial swelling, and formation of myelin-like structures. Biochemical results showed changes in the mitochondrial membrane potential, reactive oxygen species content, lipid peroxidation, and plasma membrane fluidity. In addition, the formation of autophagic vacuoles was observed. These findings indicate that BTU-1, BTU-2, and BTU-3 induced profound morphological, ultrastructural, and biochemical alterations in epimastigote forms, triggering an autophagic-dependent cell death pathway.
COVID-19: The question of genetic diversity and therapeutic intervention approaches David Livingstone Alves Figueiredo, João Paulo Bianchi Ximenez, Fábio Rodrigues Ferreira Seiva, Carolina Panis, Rafael dos Santos Bezerra, Adriano Ferrasa, Alessandra Lourenço Cecchini, Alexandra Ivo de Medeiros, Ana Marisa Fusco Almeida, Anelisa Ramão, Angelica Beate Winter Boldt, Carla Fredrichsen Moya, Chung Man Chin, Daniel de Paula, Daniel Rech, Daniela Fiori Gradia, Danielle Malheiros, Danielle Venturini, Eliandro Reis Tavares, Emerson Carraro, Enilze Maria de Souza Fonseca Ribeiro, Evani Marques Pereira, Felipe Francisco Tuon, Franciele Aní Caovilla Follador, Glaura Scantamburlo Alves Fernandes, Hélito Volpato, Ilce Mara de Syllos Cólus, Jaqueline Carvalho de Oliveira, Jean Henrique da Silva Rodrigues, Jean Leandro dos Santos, Jeane Eliete Laguila Visentainer, Juliana Cristina Brandi, Juliana Mara Serpeloni, Juliana Sartori Bonini, Karen Brajão de Oliveira, Karine Fiorentin, Léia Carolina Lucio, Ligia Carla Faccin-Galhardi, Lirane Elize Defante Ferreto, Lucy Megumi Yamauchi Lioni, Marcia Edilaine Lopes Consolaro, Marcelo Ricardo Vicari, Marcos Abdo Arbex, Marcos Pileggi, Maria Angelica Ehara Watanabe, Maria Antônia Ramos Costa, Maria José S. Mendes Giannini, Marla Karine Amarante, Najeh Maissar Khalil, Quirino Alves de Lima Neto, Roberto H. Herai, Roberta Losi Guembarovski, Rogério N. Shinsato, Rubiana Mara Mainardes, Silvana Giuliatti, Sueli Fumie Yamada-Ogatta, Viviane Knuppel de Quadros Gerber, Wander Rogério Pavanelli, Weber Claudio da Silva, Maria Luiza Petzl-Erler, Valeria Valente, Christiane Pienna Soares, Luciane Regina Cavalli, Wilson Araujo Silva Jr Genetics and Molecular Biology, 2021
Hydrogel Containing Oleoresin From Copaifera officinalis Presents Antibacterial Activity Against Streptococcus agalactiae Ana Elisa Belotto Morguette, Briani Gisele Bigotto, Renata de Lima Varella, Gabriella Maria Andriani, Laís Fernanda de Almeida Spoladori, Patrícia Moraes Lopes Pereira, Fabio Goulart de Andrade, Cesar Armando Contreras Lancheros, Celso Vataru Nakamura, Nilton Syogo Arakawa, Marcos Luciano Bruschi, José Carlos Tomaz, Audrey Alesandra Stinghen Garcia Lonni, Gilselena Kerbauy, Eliandro Reis Tavares, Lucy Megumi Yamauchi, Sueli Fumie Yamada-Ogatta Frontiers in Microbiology, 2019
Detection of OXA-58-producing Acinetobacter bereziniae in Brazil Larissa dos Santos Fávaro, Suelen Balero de Paula-Petroli, Priscila Romanin, Eliandro dos Reis Tavares, Renan Augusto Ribeiro, Mariangela Hungria, Admilton Gonçalves de Oliveira, Lucy Megumi Yamauchi, Sueli Fumie Yamada-Ogatta, Floristher Elaine Carrara-Marroni Journal of Global Antimicrobial Resistance, 2019
Phenotypic characteristics and transcriptome profile of Cryptococcus gattii biofilm Eliandro Reis Tavares, Bárbara Gionco, Ana Elisa Belotto Morguette, Gabriella Maria Andriani, Alexandre Tadachi Morey, Anderson Oliveira do Carmo, Ulisses de Pádua Pereira, Galdino Andrade, Admilton Gonçalves de Oliveira, Phileno Pinge-Filho, Celso Vataru Nakamura, Lucy Megumi Yamauchi, Sueli Fumie Yamada-Ogatta Scientific Reports, 2019
Complete genome sequence of Bacillus velezensis LABIM40, an effective antagonist of fungal plant pathogens Julia Pezarini Baptista, Paula Pinheiro Sanches, Gustavo Manoel Teixeira, Alexandre Tadachi Morey, Eliandro Reis Tavares, Sueli Fumie Yamada-Ogatta, Sérgio Paulo Dejato da Rocha, Mariangela Hungria, Renan Augusto Ribeiro, Maria Isabel Balbi-Peña, Roberta Torres Chideroli, Ulisses de Padua Pereira, Admilton Gonçalves de Oliveira Genome Announcements, 2018