Using Digital PCR to Unravel the Occurrence of Piroplasmids, <i>Bartonella</i> spp., and <i>Borrelia</i> spp. in Wild Animals from Brazil
Piroplasmids (<i>Babesia</i> spp., <i>Rangelia</i> spp., <i>Theileria</i> spp., <i>Cytauxzoon</i> spp.) are tick-borne apicomplexan protozoa that infect, depending on the species, erythrocytes and leucocytes in a wide variety of mammals and birds. The...
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2025-06-01
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| author | Ana Cláudia Calchi Anna Claudia Baumel Mongruel Fernanda Beatriz Pereira Cavalcanti Lilliane Bartone José Maurício Barbanti Duarte Emília Patrícia Medici Danilo Kluyber Mayara G. Caiaffa Mario Henrique Alves Arnaud Leonard Jean Desbiez Taciana Fernandes Souza Barbosa Coelho Rosangela Zacarias Machado Edward B. Breitschwerdt Ricardo G. Maggi Marcos Rogério André |
| author_facet | Ana Cláudia Calchi Anna Claudia Baumel Mongruel Fernanda Beatriz Pereira Cavalcanti Lilliane Bartone José Maurício Barbanti Duarte Emília Patrícia Medici Danilo Kluyber Mayara G. Caiaffa Mario Henrique Alves Arnaud Leonard Jean Desbiez Taciana Fernandes Souza Barbosa Coelho Rosangela Zacarias Machado Edward B. Breitschwerdt Ricardo G. Maggi Marcos Rogério André |
| author_sort | Ana Cláudia Calchi |
| collection | DOAJ |
| description | Piroplasmids (<i>Babesia</i> spp., <i>Rangelia</i> spp., <i>Theileria</i> spp., <i>Cytauxzoon</i> spp.) are tick-borne apicomplexan protozoa that infect, depending on the species, erythrocytes and leucocytes in a wide variety of mammals and birds. The genera <i>Bartonella</i> and <i>Borrelia</i> include vector-borne bacteria that can infect and cause disease in both animals and humans. Detection of hemotropic bacteria and piroplasmids in wild animals is often challenging due to low bacteremia or parasitemia. Digital (d)PCR has proven to be an effective modality for the detection and quantification of DNA of hemotropic pathogens with low parasitemia. This study compared dPCR results from 366 biological samples from seven different Brazilian wild animal groups (5 Xenarthra species, 5 deer species, 3 felid species, 1 canid species, 3 rodent species, 1 bat species, 1 tapir species, and 12 bird species) to two other molecular diagnostic techniques: quantitative real-time (qPCR) and nested (nPCR). For this study, DNA extracted from wild animal blood and spleen samples were subjected to a multiplex dPCR assay for piroplasmids, <i>Bartonella</i> spp., and <i>Borrelia</i> spp. For comparison, the same primers and probes for each agent were used in qPCR assays. Additionally, an nPCR based on the 18S rRNA gene for piroplasmids was performed. The proportions of positive results obtained using dPCR were 85.5% for piroplasmids, 33.6% for <i>Bartonella</i> spp., and 16.7% for <i>Borrelia</i> spp. For all tested agents, dPCR proved to be the technique with the highest sensitivity, making it a useful tool for screening vector-borne agents in biological samples from wild animals with low parasitemia. |
| format | Article |
| id | doaj-art-28cfbf1f57934a4792b00f08ead87e1c |
| institution | Kabale University |
| issn | 2076-0817 |
| language | English |
| publishDate | 2025-06-01 |
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| spelling | doaj-art-28cfbf1f57934a4792b00f08ead87e1c2025-08-20T03:29:51ZengMDPI AGPathogens2076-08172025-06-0114656710.3390/pathogens14060567Using Digital PCR to Unravel the Occurrence of Piroplasmids, <i>Bartonella</i> spp., and <i>Borrelia</i> spp. in Wild Animals from BrazilAna Cláudia Calchi0Anna Claudia Baumel Mongruel1Fernanda Beatriz Pereira Cavalcanti2Lilliane Bartone3José Maurício Barbanti Duarte4Emília Patrícia Medici5Danilo Kluyber6Mayara G. Caiaffa7Mario Henrique Alves8Arnaud Leonard Jean Desbiez9Taciana Fernandes Souza Barbosa Coelho10Rosangela Zacarias Machado11Edward B. Breitschwerdt12Ricardo G. Maggi13Marcos Rogério André14Vector-Borne Bioagents Laboratory (VBBL), Department of Pathology, Reproduction and One Health, School of Agricultural and Veterinarian Sciences (FCAV), São Paulo State University (UNESP), Jaboticabal 14884-900, SP, BrazilVector-Borne Bioagents Laboratory (VBBL), Department of Pathology, Reproduction and One Health, School of Agricultural and Veterinarian Sciences (FCAV), São Paulo State University (UNESP), Jaboticabal 14884-900, SP, BrazilPrograma de Pós-Graduação em Ciências Veterinárias, FCAV-UNESP, São Paulo State University (UNESP), Jaboticabal 14884-900, SP, BrazilIntracellular Pathogens Research Laboratory, Comparative Medicine Institute, College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27607, USADeer Research and Conservation Center (NUPECCE), São Paulo State University (UNESP), Jaboticabal 14884-900, SP, BrazilLowland Tapir Conservation Initiative (LTCI), Instituto de Pesquisas Ecológicas (IPÊ), Nazaré Paulista 12960-000, SP, BrazilICAS—Instituto de Conservacao de Animais Silvestres, Campo Grande, Mato Grosso do Sul 79040-290, MS, BrazilICAS—Instituto de Conservacao de Animais Silvestres, Campo Grande, Mato Grosso do Sul 79040-290, MS, BrazilICAS—Instituto de Conservacao de Animais Silvestres, Campo Grande, Mato Grosso do Sul 79040-290, MS, BrazilICAS—Instituto de Conservacao de Animais Silvestres, Campo Grande, Mato Grosso do Sul 79040-290, MS, BrazilSection of Arbovirology and Hemorrhagic Fevers, Coordinator of the Rabies Diagnosis Laboratory, Evandro Chagas Institute MS-SVS, São Brás, Belém 66093-020, PA, BrazilVector-Borne Bioagents Laboratory (VBBL), Department of Pathology, Reproduction and One Health, School of Agricultural and Veterinarian Sciences (FCAV), São Paulo State University (UNESP), Jaboticabal 14884-900, SP, BrazilIntracellular Pathogens Research Laboratory, Comparative Medicine Institute, College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27607, USAIntracellular Pathogens Research Laboratory, Comparative Medicine Institute, College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27607, USAVector-Borne Bioagents Laboratory (VBBL), Department of Pathology, Reproduction and One Health, School of Agricultural and Veterinarian Sciences (FCAV), São Paulo State University (UNESP), Jaboticabal 14884-900, SP, BrazilPiroplasmids (<i>Babesia</i> spp., <i>Rangelia</i> spp., <i>Theileria</i> spp., <i>Cytauxzoon</i> spp.) are tick-borne apicomplexan protozoa that infect, depending on the species, erythrocytes and leucocytes in a wide variety of mammals and birds. The genera <i>Bartonella</i> and <i>Borrelia</i> include vector-borne bacteria that can infect and cause disease in both animals and humans. Detection of hemotropic bacteria and piroplasmids in wild animals is often challenging due to low bacteremia or parasitemia. Digital (d)PCR has proven to be an effective modality for the detection and quantification of DNA of hemotropic pathogens with low parasitemia. This study compared dPCR results from 366 biological samples from seven different Brazilian wild animal groups (5 Xenarthra species, 5 deer species, 3 felid species, 1 canid species, 3 rodent species, 1 bat species, 1 tapir species, and 12 bird species) to two other molecular diagnostic techniques: quantitative real-time (qPCR) and nested (nPCR). For this study, DNA extracted from wild animal blood and spleen samples were subjected to a multiplex dPCR assay for piroplasmids, <i>Bartonella</i> spp., and <i>Borrelia</i> spp. For comparison, the same primers and probes for each agent were used in qPCR assays. Additionally, an nPCR based on the 18S rRNA gene for piroplasmids was performed. The proportions of positive results obtained using dPCR were 85.5% for piroplasmids, 33.6% for <i>Bartonella</i> spp., and 16.7% for <i>Borrelia</i> spp. For all tested agents, dPCR proved to be the technique with the highest sensitivity, making it a useful tool for screening vector-borne agents in biological samples from wild animals with low parasitemia.https://www.mdpi.com/2076-0817/14/6/567PCR techniquesdPCRhemoparasitesvector-borne agentswild animals |
| spellingShingle | Ana Cláudia Calchi Anna Claudia Baumel Mongruel Fernanda Beatriz Pereira Cavalcanti Lilliane Bartone José Maurício Barbanti Duarte Emília Patrícia Medici Danilo Kluyber Mayara G. Caiaffa Mario Henrique Alves Arnaud Leonard Jean Desbiez Taciana Fernandes Souza Barbosa Coelho Rosangela Zacarias Machado Edward B. Breitschwerdt Ricardo G. Maggi Marcos Rogério André Using Digital PCR to Unravel the Occurrence of Piroplasmids, <i>Bartonella</i> spp., and <i>Borrelia</i> spp. in Wild Animals from Brazil Pathogens PCR techniques dPCR hemoparasites vector-borne agents wild animals |
| title | Using Digital PCR to Unravel the Occurrence of Piroplasmids, <i>Bartonella</i> spp., and <i>Borrelia</i> spp. in Wild Animals from Brazil |
| title_full | Using Digital PCR to Unravel the Occurrence of Piroplasmids, <i>Bartonella</i> spp., and <i>Borrelia</i> spp. in Wild Animals from Brazil |
| title_fullStr | Using Digital PCR to Unravel the Occurrence of Piroplasmids, <i>Bartonella</i> spp., and <i>Borrelia</i> spp. in Wild Animals from Brazil |
| title_full_unstemmed | Using Digital PCR to Unravel the Occurrence of Piroplasmids, <i>Bartonella</i> spp., and <i>Borrelia</i> spp. in Wild Animals from Brazil |
| title_short | Using Digital PCR to Unravel the Occurrence of Piroplasmids, <i>Bartonella</i> spp., and <i>Borrelia</i> spp. in Wild Animals from Brazil |
| title_sort | using digital pcr to unravel the occurrence of piroplasmids i bartonella i spp and i borrelia i spp in wild animals from brazil |
| topic | PCR techniques dPCR hemoparasites vector-borne agents wild animals |
| url | https://www.mdpi.com/2076-0817/14/6/567 |
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