Advanced culture techniques, next generation sequencing, and immunoblot applied to the study of a Brazilian relapsing fever spirochete

• Grant number: 19/17960-3
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Start date: December 06, 2019
• End date: June 29, 2020
• Field of knowledge: Biological Sciences - Microbiology - Applied Microbiology
• Principal Investigator: Marcelo Bahia Labruna
• Grantee: Sebastián Alejandro Munoz Leal
• Supervisor: Job Lopez
• Host Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Institution abroad: Baylor College of Medicine, United States
• Associated to the scholarship: 18/02521-1 - Study on tick relapsing fever group spirochetes (Spirochaetaceae: Borrelia) on ticks of genus Ornithodoros (Acari: Argasidae) parasitizing humans in Brazil, BP.PD

Abstract

In South America, interest in tick-borne relapsing fever begun in the early 20th century and extended almost for 50 years. After this period, research on the disease vanished, opening a gap that has prevented a current approach with contemporary tools. Recently, its etiologic agent, the spirochete Borrelia venezuelensis, was re-isolated from its vector Ornithodoros rudis, in the Maranhão State (MA), northeastern Brazil. If we are to understand the eco-epidemiology of B. venezuelensis in this country, mainstream techniques are now required to develop studies. The objectives of the present project are (1) the standardization of in vitro culture of the isolated B. venezuelensis RMA01 stocked in our laboratory, (2) the sequencing of its genome, and (3) the design of a specific immunoblot for recognizing the circulation of B. venezuelensis antibodies in canine and human sera from MA. To this end, B. venezuelensis RMA01 will be re-activated in laboratory mice and cultivated in a modified version of the Barbour-Stoenner-Kelly (mBSK) medium. DNA extracted from mBSK-grown borreliae will be submitted to Next Generation Sequencing to obtain the complete sequences of its chromosome and large plasmid. Sequences of bipA gene will be transformed and expressed to obtain recombinant BipA protein (rBipA). Immunoblots composed by whole-cell lysates obtained from cultures and rBipA will be employed for testing anti-B. venezuelensis reactiveness of canine and human sera from MA. In its conjunct, the results of this study will represent the first efforts to understand the eco-epidemiology of tick-borne relapsing fever in Brazil. Serological tests will bring unprecedented data demonstrating whether domestic animals and/or humans are exposed to B. venezuelensis infection in MA. The genome of B. venezuelensis will provide accurate insights on the phylogenetic position of this spirochete, enabling to infer the evolutionary relationships with other species in the genus.

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)

MUNOZ-LEAL, SEBASTIAN; RAMIREZ, DIEGO G.; LUZ, HERMES R.; FACCINI, JOAO L. H.; LABRUNA, MARCELO B.. ``Candidatus Borrelia ibitipoquensis,{''} a Borrelia valaisiana-Related Genospecies Characterized from Ixodes paranaensis in Brazil. MICROBIAL ECOLOGY, v. 80, n. 3, MAY 2020. (18/02521-1, 19/17960-3)

MUNOZ-LEAL, SEBASTIAN; VENZAL, JOSE M.; JORGE, FELIPE R.; TEIXEIRA, BRUNO M.; LABRUNA, MARCELO B.. A new species of soft tick from dry tropical forests of Brazilian Caatinga. TICKS AND TICK-BORNE DISEASES, v. 12, n. 5, SEP 2021. (18/02521-1, 19/17960-3)

MUNOZ-LEAL, SEBASTIAN; MARTINS, MARIA M.; NAVA, SANTIAGO; LANDULFO, GABRIEL A.; SIMONS, SIMONE M.; RODRIGUES, VINICIUS S.; RAMOS, VANESSA N.; SUZIN, ADRIANE; SZABO, MATIAS P. J.; LABRUNA, MARCELO B.. Ornithodoros cerradoensis n. sp. (Acari: Argasidae), a member of the Ornithodoros talaje (Guerin-Meneville, 1849) group, parasite of rodents in the Brazilian Savannah. TICKS AND TICK-BORNE DISEASES, v. 11, n. 5, SEP 2020. (18/02521-1, 19/17960-3)

KNEUBEHL, ALEXANDER R.; KRISHNAVAJHALA, APARNA; LEAL, SEBASTIAN MUNOZ; REPLOGLE, ADAM J.; KINGRY, LUKE C.; BERMUDEZ, SERGIO E.; LABRUNA, MARCELO B.; LOPEZ, JOB E.. Comparative genomics of the Western Hemisphere soft tick-borne relapsing fever borreliae highlights extensive plasmid diversity. BMC Genomics, v. 23, n. 1, p. 22-pg., 2022-05-31. (19/17960-3, 18/02521-1)

PACHECO, THABATA DOS ANJOS; MUNOZ-LEAL, SEBASTIAN; MAIA, MAERLE O.; MARTINS, THIAGO F.; DE OLIVEIRA, GLAUBER M. B.; FONSECA, EDUARDO P.; PEDRONI, FERNANDO; ROSSI, ROGERIO V.; PACHECO, RICHARD C.. Molecular detection of Rickettsia spp. in ticks associated with non-volant small mammals from the Brazilian Cerrado, with notes on a divergent morphotype of Ornithodoros guaporensis. INTERNATIONAL JOURNAL OF ACAROLOGY, v. 47, n. 3, FEB 2021. (18/02521-1, 19/17960-3)

MUNOZ-LEAL, SEBASTIAN; RAMIREZ, DIEGO G.; LUZ, HERMES R.; FACCINI, JOAO L. H.; LABRUNA, MARCELO B.. "Candidatus Borrelia ibitipoquensis," a Borrelia valaisiana-Related Genospecies Characterized from Ixodes paranaensis in Brazil. MICROBIAL ECOLOGY, v. 80, n. 3, p. 8-pg., 2020-05-04. (18/02521-1, 19/17960-3)

MUNOZ-LEAL, SEBASTIAN; FACCINI-MARTINEZ, ALVARO A.; TEIXEIRA, BRUNO M.; MARTINS, MARIA MARLENE; SERPA, MARIA CAROLINA A.; OLIVEIRA, GLAUBER M. B.; JORGE, FELIPE R.; PACHECO, RICHARD C.; COSTA, FRANCISCO B.; LUZ, HERMES R.; et al. Relapsing Fever Group Borreliae in Human-Biting Soft Ticks, Brazil. Emerging Infectious Diseases, v. 27, n. 1, p. 321-324, JAN 2021. (18/02521-1, 19/17960-3)

MUNOZ-LEAL, SEBASTIAN; FACCINI-MARTINEZ, ALVARO A.; PEREZ-TORRES, JAIRO; CHALA-QUINTERO, SANDRA M.; HERRERA-SEPULVEDA, MARIA T.; CUERVO, CLAUDIA; LABRUNA, MARCELO B.. Novel Borrelia genotypes in bats from the Macaregua Cave, Colombia. ZOONOSES AND PUBLIC HEALTH, v. 68, n. 1, p. 12-18, FEB 2021. (18/02521-1, 19/17960-3)