Characterization of tick cell lines and their use as a substrate for the growth, maintenance and protein expression profile of Anaplasma marginale

• Grant number: 21/06758-9
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: January 01, 2022
• End date: June 10, 2025
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Preventive Veterinary Medicine
• Principal Investigator: Darci Moraes Barros-Battesti
• Grantee: Ana Carolina Castro Santiago
• Host Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil
• Associated scholarship(s): 23/11270-0 - Identification and characterization of Rhipicephalus microplus (RBME-6) and Amblyomma sculptum (ASE-14) tick cell transcripts in response to Anaplasma marginale infection, BE.EP.DR

Abstract

Ticks are arthropods of great medical and veterinary importance, with many species responsible for transmitting viruses, bacteria and protozoa to their hosts. Some of the tick-borne pathogens are difficult to grow in vitro, and do not grow in artificial media. Tick cell culture has been an excellent tool to isolate and study pathogens, allowing the production of antigenic material for diagnostic tests, production of antibodies and vaccines, and for studies of host-vector-pathogen relationships. In this sense, this study aims to characterize tick cell lines and use them as a substrate for the growth, maintenance and protein expression profile of Anaplasma marginale. For this, we will use primary cultures and established lineages of tick cells from Amblyomma sculptum (Berlese) and Rhipicephalus (Boophilus) microplus (Canestrini) eggs. The primary cultures will be prepared with egg masses and kept at 30 ° C in L-15B medium (Leibovitz) supplemented with Bovine Fetal Serum, subcultured when they reach the necessary confluence and cryopreserved in several passages. For the characterization of cells in culture, morphological characterization with histological techniques and karyotyping will be performed. The cells inoculated with Anaplasma marginale canis will be analyzed by slide preparations and counts of infected cells under optical microscopy, by real-time PCR and by Transmission Electron Microscopy. In addition, a differential proteomic analysis of infected and uninfected cells will be carried out, aiming at the search for proteins that may be used as vaccine antigens in the future.

Scientific publications

(15)

(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)

CASTRO-SANTIAGO, ANA CAROLINA; LIMA-DUARTE, LEIDIANE; CAMARGO, JAQUELINE VALERIA; DE ALMEIDA, BEATRIZ ROCHA; SIMONS, SIMONE MICHAELA; MATHIAS, LUIS ANTONIO; BASSINI-SILVA, RICARDO; MACHADO, ROSANGELA ZACARIAS; ANDRE, MARCOS ROGERIO; BARROS-BATTESTI, DARCI MORAES. Artificial Feeding of Ornithodoros fonsecai and O. brasiliensis (Acari: Argasidae) and Investigation of the Transstadial Perpetuation of Anaplasma marginale. MICROORGANISMS, v. 11, n. 7, p. 11-pg., 2023-07-01. (20/03190-9, 21/06758-9, 19/09915-8, 20/11755-6)

BASSINI-SILVA, RICARDO; CASTRO-SANTIAGO, ANA CAROLINA; CALCHI, ANA CLAUDIA; PERLES, LIVIA; TAKATSU, JULIA CARVALHO; ALENCAR, ISABEL D. C. C.; OCHOA, RONALD; DOWLING, ASHLEY P. G.; WERTHER, KARIN; ANDRE, MARCOS ROGERIO; et al. Sleeping with the enemy: case reports of Ornithonyssus bursa (Berlese, 1888) (Mesostigmata: Macronyssidae) causing human dermatitis in Brazil. Parasitology Research, v. 121, n. 9, p. 9-pg., 2022-07-08. (21/06758-9, 18/24667-8, 19/15150-4, 21/02295-4, 20/07826-5, 20/11755-6, 19/19853-0, 17/01416-7)

BASSINI-SILVA, RICARDO; CALCHI, ANA CLAUDIA; CASTRO-SANTIAGO, ANA CAROLINA; MAROCCO, JOAO CARLOS; DORIGONI, LETICIA; DE QUADROS, ROSILEIA MARINHO; ANDRE, MARCOS ROGERIO; BARROS-BATTESTI, DARCI MORAES; DOWLING, ASHLEY P. G.; LABRUNA, MARCELO BAHIA; et al. Molecular evidence of Wolbachia in bat-associated mite Periglischrus Iheringi Oudemans, 1902 (Mesostigmata: Spinturnicidae) from Brazil. VETERINARY RESEARCH COMMUNICATIONS, v. 49, n. 1, p. 8-pg., 2025-02-01. (20/11755-6, 24/01231-0, 21/06758-9, 20/07826-5, 22/05615-2, 19/19853-0)

BASSINI-SILVA, RICARDO; CHAGAS, MARIA EDUARDA MORAES DAS; CALCHI, ANA CLAUDIA; CASTRO-SANTIAGO, ANA CAROLINA; MUNHOZ DE MELLO, ERICA; SOARES, GABRIELLA; DORNELAS, LEORMANDO FORTUNATO; MAROCCO, JOAO CARLOS; DORIGONI, LETICIA; DOS SANTOS DA SILVA, ALLYSON; et al. Contribution to the knowledge of Periglischrus iheringi Oudemans, 1902 (Mesostigmata: Spinturnicidae) associated with Artibeus bats (Chiroptera: Phyllostomidae) from Brazil. INTERNATIONAL JOURNAL OF ACAROLOGY, v. 51, n. 1, p. 6-pg., 2025-01-04. (23/09336-3, 20/11755-6, 24/01231-0, 21/06758-9, 20/07826-5, 19/19853-0)

MELLO-OLIVEIRA, VICTOR DE SOUZA; JACINAVICIUS, FERNANDO DE CASTRO; GUERRO-DE-SOUZA, MARIANE; DORNELAS JR, LEORMANDO FORTUNATO; CASTRO-SANTIAGO, ANA CAROLINA; CALCHI, ANA CLAUDIA; LOURENCO, ELIZABETE CAPTIVO; DOWLING, ASHLEY P. G.; FAMADAS, KATIA MARIA; ANDRE, MARCOS ROGERIO; et al. Contribution to the knowledge of Parichoronyssus bakeri Morales-Malacara and Guerrero, 2007 (Mesostigmata: Macronyssidae): new locality and host-association records with additional molecular data. ACAROLOGIA, v. 64, n. 1, p. 7-pg., 2023-12-14. (20/11755-6, 18/24667-8, 23/06748-9, 20/07826-5, 19/19853-0, 23/06878-0, 22/08620-7, 17/01416-7, 21/06758-9)

CASTRO-SANTIAGO, ANA CAROLINA; LIMA-DUARTE, LEIDIANE; CAMARGO, JAQUELINE VALERIA; SIMONS, SIMONE MICHAELA; BASSINI-SILVA, RICARDO; CALCHI, ANA CLAUDIA; DE OLIVEIRA, GLAUBER MENESES BARBOZA; HORTA, MAURICIO CLAUDIO; MUNOZ-LEAL, SEBASTIAN; LABRUNA, MARCELO BAHIA; et al. New records on autogenic behavior among populations of Ornithodoros fonsecai (Labruna and Venzal, 2009) (Ixodida: Argasidae) from northeastern Brazil. VETERINARY RESEARCH COMMUNICATIONS, v. 49, n. 2, p. 9-pg., 2025-04-01. (20/11755-6, 23/07786-1, 21/06758-9, 20/07826-5, 22/05615-2)

BASSINI-SILVA, RICARDO; CALCHI, ANA CLAUDIA; CASTRO-SANTIAGO, ANA CAROLINA; SICONELLI, MARCIO JUNIO LIMA; ANDRE, MARCOS ROGERIO; BARROS-BATTESTI, DARCI MORAES; JACINAVICIUS, FERNANDO DE CASTRO. Co-infestation caused by Goniodes pavonis (L., 1758) (Ischnocera: Goniodidae) and Goniocotes rectangulatus Nitzsch [In Giebel], 1866 on white Indian peafowl, Pavo cristatus L., 1758 (Galliformes: Phasianidaede) in Brazil. VETERINARY PARASITOLOGY- REGIONAL STUDIES AND REPORTS, v. 47, p. 4-pg., 2023-11-21. (20/11755-6, 18/24667-8, 20/07826-5, 19/19853-0, 17/01416-7, 21/06758-9)

DARCI MORAES BARROS-BATTESTI; MARCOS ROGÉRIO ANDRÉ; ESTEVAM GUILHERME LUX HOPPE; GUSTAVO SERON SANCHES; RICARDO BASSINI-SILVA; ANA CLAÚDIA CALCHI; LÍVIA OLIVEIRA ANDRADE; PATRÍCIA PARREIRA PERIN; THIAGO FERNANDES MARTINS; ANA CAROLINA CASTRO-SANTIAGO; et al. Interessantes registros dos carrapatos Ornithodoros rostratus e Amblyomma sculptum parasitando Pteronura brasiliensis na região centro-oeste do Brasil, com informações sobre a investigação de patógenos. Revista Brasileira de Parasitologia Veterinária, v. 33, n. 1, 2023-12-22. (21/06758-9, 20/05987-1, 17/01416-7, 19/03167-0, 20/11755-6, 19/19853-0, 20/07826-5, 18/24667-8)

CALCHI, ANA CLAUDIA; BRAGA, LAIZA DE QUEIROZ VIANA; BASSINI-SILVA, RICARDO; CASTRO-SANTIAGO, ANACAROLINA; HERRERA, HEITOR MIRAGLIA; SOARES, JOAO FABIO; BARROS-BATTESTI, DARCI MORAES; MACHADO, ROSANGELA ZACARIAS; ROCHA, FABIANA LOPES; ANDRE, MARCOS ROGERIO. Phylogenetic inferences based on distinct molecular markers reveals a novel Babesia ( Babesia pantanalensis nov. sp.) and a Hepatozoon americanum-related genotype in crab-eating foxes ( Cerdocyon thous ). Experimental Parasitology, v. 262, p. 10-pg., 2024-05-22. (20/11755-6, 20/12037-0, 20/07826-5, 21/06758-9, 22/08543-2)

CALCHI, ANA CLAUDIA; DUARTE, JOSE MAURICIO BARBANTI; CASTRO-SANTIAGO, ANA CAROLINA; BASSINI-SILVA, RICARDO; BARROS-BATTESTI, DARCI MORAES; MACHADO, ROSANGELA ZACARIAS; ANDRE, MARCOS ROGERIO. Genetic diversity of Theileria spp. in deer (Artiodactyla: Cervidae) from Brazil. Parasitology Research, v. 123, n. 11, p. 17-pg., 2024-11-01. (20/11755-6, 20/12037-0, 20/07826-5, 21/06758-9, 22/08543-2)

BASSINI-SILVA, RICARDO; DAS CHAGAS, MARIA EDUARDA MORAES; MELLO-OLIVEIRA, VICTOR DE SOUZA; CALCHI, ANA CLAUDIA; CASTRO-SANTIAGO, ANA CAROLINA; ANDRADE, LIVIA DE OLIVEIRA; BENEDET, GABRIELA COELHO; PEREIRA, FERNANDA MARA ARAGAO MACEDO; SOARES-NETO, LAURO LEITE; HIPPOLITO, ALICIA GIOLO; et al. Eutrichophilus cordiceps Mjoberg, 1910 (Ischnocera: Trichodectidae) in Spiny Tree Porcupines (Coendou villosus): New locality records and the first molecular evidence of association with Bartonella sp. Parasitology International, v. 100, p. 7-pg., 2024-06-01. (18/24667-8, 20/07826-5, 19/19853-0, 23/06878-0, 23/09336-3, 17/01416-7, 21/06758-9)

BARROS-BATTESTI, DARCI MORAES; ANDRE, MARCOS ROGERIO; HOPPE, ESTEVAM GUILHERME LUX; SERON, GUSTAVO; BASSINI-SILVA, RICARDO; CALCHI, ANA CLAUDIA; ANDRADE, LIVIA OLIVEIRA; PERIN, PATRICIA PARREIRA; MARTINS, THIAGO FERNANDES; CASTRO-SANTIAGO, ANA CAROLINA; et al. Noteworthy records of the ticks Ornithodoros rostratus and Amblyomma sculptum parasitizing Pteronura brasiliensis in the central-western region of Brazil, with pathogen investigation notes. REVISTA BRASILEIRA DE PARASITOLOGIA VETERINARIA, v. 33, n. 1, p. 7-pg., 2024-01-01. (20/11755-6, 18/24667-8, 20/05987-1, 20/07826-5, 19/19853-0, 19/03167-0, 17/01416-7, 21/06758-9)

BASSINI-SILVA, RICARDO; ANDRE, MARCOS ROGERIO; PEREIRA, ALICE; DAS CHAGAS, MARIA EDUARDA MORAES; MELLO-OLIVEIRA, VICTOR DE SOUZA; CALCHI, ANA CLAUDIA; MONGRUEL, ANNA CLAUDIA BAUMEL; CASTRO-SANTIAGO, ANA CAROLINA; MACHADO, DALIA MONIQUE RIBEIRO; BARROS-BATTESTI, DARCI MORAES; et al. First molecular detection of Bartonella spp. in two chewing louse species (Amblycera: Menoponidae and Ischnocera: Philopteridae) parasitizing Calonectris borealis (Cory, 1881) (Procellariiformes: Procellariidae). VETERINARY PARASITOLOGY- REGIONAL STUDIES AND REPORTS, v. 58, p. 5-pg., 2025-02-01. (23/09336-3, 20/11755-6, 19/26403-0, 21/06758-9, 20/07826-5, 22/05615-2, 19/19853-0)

BASSINI-SILVA, RICARDO; CASTRO-SANTIAGO, ANA CAROLINA; CALCHI, ANA CLAUDIA; DE MELLO, ERICA M.; DORNELAS JR, LEORMANDO FORTUNATO; VIANA, AMANDA DE OLIVEIRA; CUNHA, IRINEU NORBERTO; SABINO-SANTOS, GILBERTO; DA SILVA, ALLYSON DOS SANTOS; TELINO-JUNIOR, WALLACE RODRIGUES; et al. A catalogue of spinturnicid species (Mesostigmata: Spinturnicidae) from Brazil: distribution, host-specificity, and new locality records. INTERNATIONAL JOURNAL OF ACAROLOGY, v. 50, n. 3, p. 15-pg., 2024-02-21. (18/24667-8, 22/05615-2, 19/19853-0, 17/01416-7, 21/06758-9, 11/06810-9)

BASSINI-SILVA, RICARDO; MELLO-OLIVEIRA, VICTOR DE SOUZA; DE ALMEIDA, BEATRIZ ROCHA; PESENATO, ISABELLA PEREIRA; CALCHI, ANA CLAUDIA; CASTRO-SANTIAGO, ANA CAROLINA; LOURENCO, ELIZABETE CAPTIVO; FAMADAS, KATIA MARIA; WELBOURN, CAL; OCHOA, RONALD; et al. Molecular detection of Ehrlichia Sp. in bat-associated chigger mites (trombidiformes: leeuwenhoekiidae and trombiculidae S. s.) from Brazil. Experimental and Applied Acarology, v. 94, n. 3, p. 9-pg., 2025-04-01. (22/05615-2, 20/07826-5, 22/11576-0, 23/09381-9, 19/19853-0, 24/01231-0, 23/06878-0, 21/06758-9, 20/11755-6)