Successful Infection of Tick Cell Cultures of Rhipicephalus sanguineus (Tropical Lineage) with Ehrlichia canis

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dc.contributorLab. Parasitologiapt_BR
dc.contributor(LDI) Laboratório de Desenvolvimento e Inovação Industrialpt_BR
dc.contributor.authorBarros-Battesti, Darci Moraespt_BR
dc.contributor.authorMachado, Rosangela Zacariaspt_BR
dc.contributor.authorAndre, Marcos Rogeriopt_BR
dc.contributor.authorMarques de Sousa, Keyla Carstenspt_BR
dc.contributor.authorFranze, Daniella Aparecidapt_BR
dc.contributor.authorDuarte, Leidiane Limapt_BR
dc.contributor.authorCirelli-Moraes, Angelinapt_BR
dc.contributor.authorNunes, Pablo Henriquept_BR
dc.contributor.authorLabruna, Marcelo Bahiapt_BR
dc.contributor.authorMoraes-Filho, Jonaspt_BR
dc.contributor.authorMartins, Maria Marlenept_BR
dc.contributor.authorJuan Szabo, Matias Pablopt_BR
dc.identifier.citationBarros-Battesti DM, Machado RZ, Andre MR, Marques de Sousa KC, Franze DA, Duarte LL, et al. Successful Infection of Tick Cell Cultures of Rhipicephalus sanguineus (Tropical Lineage) with Ehrlichia canis. Vector-Borne Zoonotic Dis.. 2018 Set;18(2):653-662. doi:10.1089/vbz.2017.2197.pt_BR
dc.description.abstractThere are two distinct lineages of ticks, Rhipicephalus sanguineus, in South America: tropical and temperate lineages. Only the tropical lineage is recognized as competent vector for Ehrlichia canis. The epidemiological data of canine monocytic ehrlichiosis is congruent with the distribution of the two lineages of R. sanguineus. Herein, we report the infection of R. sanguineus (tropical lineage) cell cultures with E. canis, after cryopreservation. R. sanguineus (tropical lineage) cell identity was confirmed by sequencing using a 16S rDNA gene fragment. Tick cell cultures were prepared in L-15B medium supplemented with 10%, 15%, and 20% Fetal Bovine Serum (FBS), and 10% of Tryptose Phosphate Broth (TPB). Cell cultures developed better at the concentration of 20% of FBS. Cultures in the fifth harvest (approximately 7 months later) were selected for the first infections. Optimal R. sanguineus cell growth and adhesion was observed (5.0?×?106 cells/mL, and the population doubling time every 57?h). Once infected with E. canis, the cultures were maintained in L-15B medium supplemented with 2% and 5% of FBS fortified with iron and 10% TPB. Infected cells were also cryopreserved. DNA was extracted from infected and noninfected cells and analyzed using quantitative real-time PCR targeting the E. canis-dsb gene. Primary culture of the fifth passage was infected by E. canis and it maintained the pathogen for at least 40 days before partial cell destruction. Subcultures of infected cells (fresh and cryopreserved cultures) onto new tick cell cultures were successful. The E. canis infection was confirmed by real-time PCR and light and transmission electron microscopy. The R. sanguineus (tropical lineage) cells infected with E. canis successfully infected new tick cell cultures, showing that these cells could be an alternative substrate for maintenance of this pathogen.pt_BR
dc.description.sponsorship(FAPESP) Fundação de Amparo à Pesquisa do Estado de São Paulopt_BR
dc.description.sponsorship(CNPq) Conselho Nacional de Desenvolvimento Científico e Tecnológicopt_BR
dc.relation.ispartofVector-Borne and Zoonotic Diseasespt_BR
dc.titleSuccessful Infection of Tick Cell Cultures of Rhipicephalus sanguineus (Tropical Lineage) with Ehrlichia canispt_BR
dc.contributor.external(UNILA) Universidade Federal da Integração Latino-Americanapt_BR
dc.contributor.external(USP) Universidade de São Paulopt_BR
dc.contributor.external(UNISA) Universidade Santo Amaropt_BR
dc.contributor.external(UFU) Universidade Federal de Uberlândiapt_BR
dc.subject.keywordbrown dog tickpt_BR
dc.subject.keywordtropical lineagept_BR
dc.subject.keywordtick cell culturept_BR
dc.subject.keywordEhrlichia canispt_BR
dc.relation.ispartofabbreviatedVector-Borne Zoonotic Dispt_BR
dc.identifier.citationabntv. 18, n. 12, p. 653-662, set. 2018pt_BR
dc.identifier.citationvancouver2018 Set;18(2):653-662pt_BR
dc.contributor.butantanFranze, Daniella Aparecida|:Aluno|:Lab. Parasitologia|:pt_BR
dc.contributor.butantanDuarte, Leidiane Lima|:Aluno|:Lab. Parasitologia|:pt_BR
dc.contributor.butantanBarros-Battesti, Darci Moraes|:Pesquisador|:Lab. Parasitologia|:PrimeiroAutor:Autor de correspondênciapt_BR
dc.contributor.butantanCirelli-Moraes, Angelina|:Aluno|:Lab. Parasitologia:Lab. Biologia Molecular|:pt_BR
dc.sponsorship.butantan(CNPq) Conselho Nacional de Desenvolvimento Científico e Tecnológico¦¦312331/2013-4pt_BR
dc.sponsorship.butantan(FAPESP) Fundação de Amparo à Pesquisa do Estado de São Paulo¦¦2007/57749-2pt_BR
dc.sponsorship.butantan(FAPESP) Fundação de Amparo à Pesquisa do Estado de São Paulo¦¦2015/26209-9pt_BR
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