Mem Inst Oswaldo Cruz, Rio de Janeiro, VOLUME 121 | 2026
Research Articles

Blechomonas campbelli from Ctenocephalides felis: in vitro development, thermotolerance, and phylogenetic insights

Letícia Gracielle Tôrres de Miranda Estevam1, Alexei Yu Kostygov2,3,+, Felipe Dutra-Rêgo1,+, Anna Luiza Moreira Martins1, Marina Andrade Freire1, Ana Cristina Vianna Mariano da Rocha Lima1, Daniel Moreira de Avelar1, Gregório Guilherme Almeida4, José Dilermando Andrade Filho1, Vyacheslav Yurchenko2, Gustavo Fontes Paz1

1Fundação Oswaldo Cruz-Fiocruz, Instituto René Rachou, Belo Horizonte, MG, Brasil1Fundação Oswaldo Cruz-Fiocruz, Instituto René Rachou, Belo Horizonte, MG, Brasil
2University of Ostrava, Faculty of Science, Life Science Research Centre, Ostrava, Czechia
3Russian Academy of Sciences, Zoological Institute, St. Petersburg, Russia
4Centro de Tecnologia de Vacinas, Belo Horizonte, MG, Brasil

DOI: 10.1590/0074-02760250286
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ABSTRACT

BACKGROUND Blechomonas spp. are flea-specific monoxenous trypanosomatids, whose biology, host range, and geographic distribution remain poorly understood, even for species inhabiting ubiquitous cat and dog fleas.
OBJECTIVES To isolate and characterise blechomonads from fleas of domestic dogs in Brazil, focusing on culture growth, morphology, and phylogenetic relationships.
METHODS Fleas collected from dogs in Sabará, Brazil, were morphologically identified and screened by cultivation. The isolate FL-1 was analysed for temperature-dependent growth, morphology, and phylogeny using 18S rRNA and gGAPDH sequences, including those mined from public metagenomic datasets.
FINDINGS Of 340 fleas (four per pool), only one culture tested positive. Growth was optimal at 25ºC, reduced at 30ºC, and absent at 35ºC. Four cell types were revealed, including a predominant “uromonad” stage specialised for attachment and aggregation. Phylogenetic analyses placed the isolate within Blechomonas campbelli and indicated that this species and Blechomonas lauriereadi occur in Ctenocephalides spp. from multiple continents.
MAIN CONCLUSIONS Blechomonas campbelli exhibits developmental adaptations to the flea gut and limited tolerance to elevated temperatures, constraining its potential to persist in warm-blooded hosts. These findings highlight the value of combining culture-based and in silico approaches to investigate flea-associated trypanosomatids.

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Financial support: This study was supported by the European Union (EU) Operational Programme Just Transition (LERCO CZ.10.03.01/00/22_003/0000003 to VY) and by the State Assignment for the Zoological Institute RAS No. 125012800894-6.
LGTME was supported by fellowships from the FAPEMIG and CAPES.
LGTME, AYK and FD-R contributed equally to this work.
+ Corresponding authors: kostygov@gmail.com | ORCID https://orcid.org/0000-0002-1516-437X; felipedutra04@hotmail.com | ORCID https://orcid.org/0000-0002-2799-8267
Received 14 October 2025
Accepted 11 May 2026

HOW TO CITE
Estevam LGTM, Kostygov AY, Dutra-Rêgo F, Martins ALM, Freire MA, Lima ACVMR, et al. Blechomonas campbelli from Ctenocephalides felis: in vitro development, thermotolerance, and phylogenetic insights. Mem Inst Oswaldo Cruz. 2026; 121: e250286.
Handling editor: Geovane Dias-Lopes | ORCID https://orcid.org/0000-0003-4448-1398

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