Ticks as potential vectors of Mycobacterium leprae: Use of tick cell lines to culture the bacilli and generate transgenic strains.

Leprosy is an infectious disease caused by Mycobacterium leprae and frequently resulting in irreversible deformities and disabilities. Ticks play an important role in infectious disease transmission due to their low host specificity, worldwide distribution, and the biological ability to support transovarial transmission of a wide spectrum of pathogens, including viruses, bacteria and protozoa. To investigate a possible role for ticks as vectors of leprosy, we assessed transovarial transmission of M. leprae in artificially-fed adult female Amblyomma sculptum ticks, and infection and growth of M. leprae in tick cell lines. Our results revealed M. leprae RNA and antigens persisting in the midgut and present in the ovaries of adult female A. sculptum at least 2 days after oral infection, and present in their progeny (eggs and larvae), which demonstrates the occurrence of transovarial transmission of this pathogen. Infected tick larvae were able to inoculate viable bacilli during blood-feeding on a rabbit. Moreover, following inoculation with M. leprae, the Ixodes scapularis embryo-derived tick cell line IDE8 supported a detectable increase in the number of bacilli for at least 20 days, presenting a doubling time of approximately 12 days. As far as we know, this is the first in vitro cellular system able to promote growth of M. leprae. Finally, we successfully transformed a clinical M. leprae isolate by inserting the reporter plasmid pCHERRY3; transformed bacteria infected and grew in IDE8 cells over a 2-month period. Taken together, our data not only support the hypothesis that ticks may have the potential to act as a reservoir and/or vector of leprosy, but also suggest the feasibility of technological development of tick cell lines as a tool for large-scale production of M. leprae bacteria, as well as describing for the first time a method for their transformation

Characterization of two strains of Anaplasma marginale isolated from cattle in Rio de Janeiro, Brazil, after propagation in tick cell culture

IDE8 tick cell cultures have been used for the isolation and propagation of several isolates of Anaplasma marginale. The genetic heterogeneity of A. marginale strains in cattle is diverse in endemic regions worldwide and the analyses of msp1α (major surface protein 1 alpha) gene sequences have allowed the identification of different strains. This study reports the isolation and propagation of two new isolates of A. marginale in IDE8 cells from blood of two cattle and their morphological and molecular characterization using light microscopy and the msp1α gene, respectively. Small colonies were observed in cytospin smears of each of the isolates 60 days after culture initiation. Based on msp1α sequence variation, the two isolates were found to be separate strains and were named AmRio1 and AmRio2. Analysis of msp1α microsatellite in both strains resulted in a single genotype, genotype E. The amino acid sequence of one MSP1α tandem repeat from the strain AmRio1 resulted in a new sequence (named 162) with one amino acid change. The results of these phylogenetic analyses demonstrated that A. marginale strains from Brazil and Argentina formed two large clusters of which one was less divergent that the other.To the National Council for Scientific and Technological Development (CNPq), and to the Coordination Office for Improvement of Higher-Education Staff (CAPES), for their financial support.Peer Reviewe

Characterization of two strains of <em>Anaplasma marginale</em> isolated from cattle in Rio de Janeiro, Brazil, after propagation in tick cell culture.

Contribution: Sequence and phylogenetic analysis.International audienceIDE8 tick cell cultures have been used for the isolation and propagation of several isolates of Anaplasma marginale. The genetic heterogeneity of A. marginale strains in cattle is diverse in endemic regions worldwide and the analyses of msp1˛ (major surface protein 1 alpha) gene sequences have allowed the identification of different strains. This study reports the isolation and propagation of two new isolates of A. marginale in IDE8 cells from blood of two cattle and their morphological and molecular characterization using light microscopy and the msp1˛ gene, respectively. Small colonies were observed in cytospin smears of each of the isolates 60 days after culture initiation. Based on msp1˛ sequence variation, the two isolates were found to be separate strains and were named AmRio1 and AmRio2. Analysis of msp1˛ microsatellite in both strains resulted in a single genotype, genotype E. The amino acid sequence of one MSP1 tandem repeat from the strain AmRio1 resulted in a new sequence (named 162) with one amino acid change. The results ofthese phylogenetic analyses demonstrated that A. marginale strains from Brazil and Argentina formed two large clusters of which one was less divergent that the other

Alterações bioquímicas, anatômicas e histopatológicas em fígado de Gallus gallus Linnaeus, 1758 experimentalmente infectados por Borrelia anserina Sakharoff, 1891

Resumo: A espiroquetose aviária é uma enfermidade septicêmica de curso agudo, cosmopolita, que acomete diversas espécies aviárias, causada por Borrelia anserina e transmitida pelo carrapato Argas miniatus. O experimento teve como objetivos avaliar as alterações bioquímicas e anátomo-histopatológicas no fígado de Gallus gallus, causadas pela infecção experimental por B. anserina. Quarenta aves da espécie G. gallus foram divididas em quatro grupos inteiramente casualizados com 10 animais cada: G1 - inoculado com soro infectado com B. anserina; G2 - inoculado com soro fisiológico a 0,9%; G3 - exposto a ninfas de terceiro ínstar de A. miniatus infectados por B. anserina; G4 - exposto a ninfas de terceiro ínstar de A. miniatus livres de B. anserina. As aves dos Grupos 1 e 3 manifestaram no 3º e 6º dias pós-inoculação (DPI) respectivamente, sintomatologia característica da doença como inapetência, perda de peso, sonolência, diarreia esverdeada, mucosas hipocoradas, penas arrepiadas e hipertermia. Os níveis de ALT do Grupo 1 mostraram-se significativamente mais elevados apenas no 12ºDPI e 24ºDPI em relação ao seu grupo controle (Grupo 2) e no Grupo 3 esses níveis se mantiveram elevados até o 20º DPI em comparação ao seu grupo controle (Grupo 4). Os níveis da enzima AST pouco oscilaram nos grupos experimentais, embora tenham sido encontradas elevações no 12ºDPI nos Grupos 1 e 3. Os fígados das aves dos Grupos 1 e 3 apresentaram à necropsia, moderada hepatomegalia, congestão, superfície irregular e coloração vermelha a cianótica; constataram-se ainda pequenos pontos esbranquiçados na superfície. A histopatologia do fígado revelou congestão, infiltrados inflamatórios mononucleares, focos de necrose fibrinoide, dilatação dos sinusoides e vacuolização de hepatócitos. A coloração de Warthin-Starry revelou, nos fígados das aves dos Grupos 1 e 3, a presença de espiroquetas compatíveis com B. anserina, frequentemente no interior de vasos sanguíneos

The taxonomic impediment: a shortage of taxonomists, not the lack of technical approaches.

Engel, Michael S, Ceríaco, Luis M P, Daniel, Gimo M, Dellapé, Pablo M, Löbl, Ivan, Marinov, Milen, Reis, Roberto E, Young, Mark T, Dubois, Alain, Agarwal, Ishan, Lehmann A., Pablo, Alvarado, Mabel, Alvarez, Nadir, Andreone, Franco, Araujo-Vieira, Katyuscia, Ascher, John S, Baêta, Délio, Baldo, Diego, Bandeira, Suzana A, Barden, Phillip, Barrasso, Diego A, Bendifallah, Leila, Bockmann, Flávio A, Böhme, Wolfgang, Borkent, Art, Brandão, Carlos R F, Busack, Stephen D, Bybee, Seth M, Channing, Alan, Chatzimanolis, Stylianos, Christenhusz, Maarten J M, Crisci, Jorge V, D'elía, Guillermo, Da Costa, Luis M, Davis, Steven R, De Lucena, Carlos Alberto S, Deuve, Thierry, Fernandes Elizalde, Sara, Faivovich, Julián, Farooq, Harith, Ferguson, Adam W, Gippoliti, Spartaco, Gonçalves, Francisco M P, Gonzalez, Victor H, Greenbaum, Eli, Hinojosa-Díaz, Ismael A, Ineich, Ivan, Jiang, Jianping, Kahono, Sih, Kury, Adriano B, Lucinda, Paulo H F, Lynch, John D, Malécot, Valéry, Marques, Mariana P, Marris, John W M, Mckellar, Ryan C, Mendes, Luis F, Nihei, Silvio S, Nishikawa, Kanto, Ohler, Annemarie, Orrico, Victor G D, Ota, Hidetoshi, Paiva, Jorge, Parrinha, Diogo, Pauwels, Olivier S G, Pereyra, Martín O, Pestana, Lueji B, Pinheiro, Paulo D P, Prendini, Lorenzo, Prokop, Jakub, Rasmussen, Claus, Rödel, Mark-Oliver, Rodrigues, Miguel Trefaut, Rodríguez, Sara M, Salatnaya, Hearty, Sampaio, Íris, Sánchez-García, Alba, Shebl, Mohamed A, Santos, Bruna S, Solórzano-Kraemer, Mónica M, Sousa, Ana C A, Stoev, Pavel, Teta, Pablo, Trape, Jean-François, Dos Santos, Carmen Van-Dúnem, Vasudevan, Karthikeyan, Vink, Cor J, Vogel, Gernot, Wagner, Philipp, Wappler, Torsten, Ware, Jessica L, Wedmann, Sonja, Zacharie, Chifundera Kusamba (2021): EDITORIAL The taxonomic impediment: a shortage of taxonomists, not the lack of technical approaches. Zoological Journal of the Linnean Society 193 (2): 381-387, DOI: 10.1093/zoolinnean/zlab072, URL: https://academic.oup.com/zoolinnean/article/193/2/381/637438