Molecular detection and genetic diversity of Bartonella species in large ruminants and associated ectoparasites from the Brazilian Cerrado

Luiz Ricardo Gonçalves; Shimon Harrus; Ricardo Gutiérrez; Heitor Miraglia Herrera; Inalda Angélica de Souza Ramos; Grasiela Edith de Oliveira Porfírio; Yaarit Nachum-Biala; Keyla Carstens Marques de Sousa; Thiago Merighi Vieira da Silva; João Bosco Vilela Campos; Wagner Lemos; Darci Moraes Barros-Battesti; Rosangela Zacarias Machado; Marcos Rogério André

doi:10.1111/tbed.13517

Molecular detection and genetic diversity of Bartonella species in large ruminants and associated ectoparasites from the Brazilian Cerrado

Luiz Ricardo Gonçalves, Shimon Harrus, Ricardo Gutiérrez, Heitor Miraglia Herrera, Inalda Angélica de Souza Ramos, Grasiela Edith de Oliveira Porfírio, Yaarit Nachum-Biala, Keyla Carstens Marques de Sousa, Thiago Merighi Vieira da Silva, João Bosco Vilela Campos, Wagner Lemos, Darci Moraes Barros-Battesti, Rosangela Zacarias Machado, Marcos Rogério André^*

^*Corresponding author for this work

The Koret School of Veterinary Medicine

Research output: Contribution to journal › Article › peer-review

6 Scopus citations

Abstract

Currently, five Bartonella species and an expanding number of Candidatus Bartonella species have globally been reported in ruminants. Likewise, different Bartonella genotypes were identified. However, studies relating to ruminant-associated Bartonella in Brazil are scarce. The current study aimed to assess the prevalence and genetic diversity of Bartonella in cattle, buffaloes and associated ectoparasites in Brazil. For this purpose, EDTA-blood samples from 75 cattle and 101 buffaloes were sampled. Additionally, 128 Rhipicephalus microplus and one Amblyomma sculptum ticks collected from cattle, and 197 R. microplus, one A. sculptum and 170 lice (Haematopinus tuberculatus) collected from buffaloes were included. Bartonella DNA was initially screened through an HRM real-time PCR assay targeting the 16S–23S internal transcribed spacer (ITS), and the positive samples were submitted to an additional HRM assay targeting the ssrA gene. The HRM-positive amplicons were sequenced, and the nucleotide identity was assessed by BLASTn. Bartonella spp.-positive DNA samples were analysed by conventional PCR assays targeting the gltA and rpoB genes, and then, the samples were cloned. Finally, the phylogenetic positioning and the genetic diversity of clones were assessed. Overall, 21 of 75 (28%) cattle blood samples and 13 of 126 (10.3%) associated ticks were positive for Bartonella bovis. Out of 101 buffaloes, 95 lice and 188 tick DNA samples, one (1%) buffalo and four (4.2%) lice were positive for Bartonella spp. Conversely, none of the ticks obtained from buffaloes were positive for Bartonella. The Bartonella sequences from buffaloes showed identity ranging from 100% (ITS and gltA) to 94% (ssrA) with B. bovis. In contrast, the Bartonella DNA sequences from lice were identical (100%) to uncultured Bartonella sp. detected in cattle tail louse (Haematopinus quadripertusus) from Israel in all amplified genes. The present study demonstrates the prevalence of new B. bovis genotypes and a cattle lice-associated Bartonella species in large ruminants and their ectoparasites from Brazil. These findings shed light on the distribution and genetic diversity of ruminant- and ectoparasite-related Bartonella in Brazil.

Original language	American English
Pages (from-to)	1888-1897
Number of pages	10
Journal	Transboundary and Emerging Diseases
Volume	67
Issue number	5
DOIs	https://doi.org/10.1111/tbed.13517
State	Published - 1 Sep 2020

Bibliographical note

Funding Information:
The authors thank the Graduate Agricultural Microbiology Program (Universidade Estadual Paulista, UNESP, Campus Jaboticabal), the FAPESP (Foundation for Research Support of the State of São Paulo—Process 2018/02753‐0), FUNDECT (Foundation for Support to the Development of Education, Science and Technology of the State of Mato Grosso do Sul, Case 59/300.187/2016) and CNPq for the Productivity Grant granted to the MRA (CNPq Process No. 302420/2017‐7). The present study was carried out with the support of the Coordination for the Improvement of Higher Education (CAPES—Code 1). L.R.G. received a ‘Sandwich’ scholarship from the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior.

Funding Information:
The authors thank the Graduate Agricultural Microbiology Program (Universidade Estadual Paulista, UNESP, Campus Jaboticabal), the FAPESP (Foundation for Research Support of the State of São Paulo—Process 2018/02753-0), FUNDECT (Foundation for Support to the Development of Education, Science and Technology of the State of Mato Grosso do Sul, Case 59/300.187/2016) and CNPq for the Productivity Grant granted to the MRA (CNPq Process No. 302420/2017-7). The present study was carried out with the support of the Coordination for the Improvement of Higher Education (CAPES—Code 1). L.R.G. received a ‘Sandwich’ scholarship from the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior.

Publisher Copyright:
© 2020 Blackwell Verlag GmbH

Keywords

Haematopinus tuberculatus
Rhipicephalus microplus
bartonellosis
buffaloes
cattle
genetic diversity

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10.1111/tbed.13517

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Gonçalves, L. R., Harrus, S., Gutiérrez, R., Herrera, H. M., de Souza Ramos, I. A., Porfírio, G. E. D. O., Nachum-Biala, Y., de Sousa, K. C. M., da Silva, T. M. V., Campos, J. B. V., Lemos, W., Moraes Barros-Battesti, D., Machado, R. Z., & André, M. R. (2020). Molecular detection and genetic diversity of Bartonella species in large ruminants and associated ectoparasites from the Brazilian Cerrado. Transboundary and Emerging Diseases, 67(5), 1888-1897. https://doi.org/10.1111/tbed.13517

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title = "Molecular detection and genetic diversity of Bartonella species in large ruminants and associated ectoparasites from the Brazilian Cerrado",

abstract = "Currently, five Bartonella species and an expanding number of Candidatus Bartonella species have globally been reported in ruminants. Likewise, different Bartonella genotypes were identified. However, studies relating to ruminant-associated Bartonella in Brazil are scarce. The current study aimed to assess the prevalence and genetic diversity of Bartonella in cattle, buffaloes and associated ectoparasites in Brazil. For this purpose, EDTA-blood samples from 75 cattle and 101 buffaloes were sampled. Additionally, 128 Rhipicephalus microplus and one Amblyomma sculptum ticks collected from cattle, and 197 R. microplus, one A. sculptum and 170 lice (Haematopinus tuberculatus) collected from buffaloes were included. Bartonella DNA was initially screened through an HRM real-time PCR assay targeting the 16S–23S internal transcribed spacer (ITS), and the positive samples were submitted to an additional HRM assay targeting the ssrA gene. The HRM-positive amplicons were sequenced, and the nucleotide identity was assessed by BLASTn. Bartonella spp.-positive DNA samples were analysed by conventional PCR assays targeting the gltA and rpoB genes, and then, the samples were cloned. Finally, the phylogenetic positioning and the genetic diversity of clones were assessed. Overall, 21 of 75 (28%) cattle blood samples and 13 of 126 (10.3%) associated ticks were positive for Bartonella bovis. Out of 101 buffaloes, 95 lice and 188 tick DNA samples, one (1%) buffalo and four (4.2%) lice were positive for Bartonella spp. Conversely, none of the ticks obtained from buffaloes were positive for Bartonella. The Bartonella sequences from buffaloes showed identity ranging from 100% (ITS and gltA) to 94% (ssrA) with B. bovis. In contrast, the Bartonella DNA sequences from lice were identical (100%) to uncultured Bartonella sp. detected in cattle tail louse (Haematopinus quadripertusus) from Israel in all amplified genes. The present study demonstrates the prevalence of new B. bovis genotypes and a cattle lice-associated Bartonella species in large ruminants and their ectoparasites from Brazil. These findings shed light on the distribution and genetic diversity of ruminant- and ectoparasite-related Bartonella in Brazil.",

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note = "Funding Information: The authors thank the Graduate Agricultural Microbiology Program (Universidade Estadual Paulista, UNESP, Campus Jaboticabal), the FAPESP (Foundation for Research Support of the State of S{\~a}o Paulo—Process 2018/02753‐0), FUNDECT (Foundation for Support to the Development of Education, Science and Technology of the State of Mato Grosso do Sul, Case 59/300.187/2016) and CNPq for the Productivity Grant granted to the MRA (CNPq Process No. 302420/2017‐7). The present study was carried out with the support of the Coordination for the Improvement of Higher Education (CAPES—Code 1). L.R.G. received a {\textquoteleft}Sandwich{\textquoteright} scholarship from the Coordena{\c c}{\~a}o de Aperfei{\c c}oamento de Pessoal de N{\'i}vel Superior. Funding Information: The authors thank the Graduate Agricultural Microbiology Program (Universidade Estadual Paulista, UNESP, Campus Jaboticabal), the FAPESP (Foundation for Research Support of the State of S{\~a}o Paulo—Process 2018/02753-0), FUNDECT (Foundation for Support to the Development of Education, Science and Technology of the State of Mato Grosso do Sul, Case 59/300.187/2016) and CNPq for the Productivity Grant granted to the MRA (CNPq Process No. 302420/2017-7). The present study was carried out with the support of the Coordination for the Improvement of Higher Education (CAPES—Code 1). L.R.G. received a {\textquoteleft}Sandwich{\textquoteright} scholarship from the Coordena{\c c}{\~a}o de Aperfei{\c c}oamento de Pessoal de N{\'i}vel Superior. Publisher Copyright: {\textcopyright} 2020 Blackwell Verlag GmbH",

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doi = "10.1111/tbed.13517",

language = "American English",

volume = "67",

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Gonçalves, LR, Harrus, S, Gutiérrez, R, Herrera, HM, de Souza Ramos, IA, Porfírio, GEDO, Nachum-Biala, Y, de Sousa, KCM, da Silva, TMV, Campos, JBV, Lemos, W, Moraes Barros-Battesti, D, Machado, RZ & André, MR 2020, 'Molecular detection and genetic diversity of Bartonella species in large ruminants and associated ectoparasites from the Brazilian Cerrado', Transboundary and Emerging Diseases, vol. 67, no. 5, pp. 1888-1897. https://doi.org/10.1111/tbed.13517

TY - JOUR

T1 - Molecular detection and genetic diversity of Bartonella species in large ruminants and associated ectoparasites from the Brazilian Cerrado

AU - Gonçalves, Luiz Ricardo

AU - Harrus, Shimon

AU - Gutiérrez, Ricardo

AU - Herrera, Heitor Miraglia

AU - de Souza Ramos, Inalda Angélica

AU - Porfírio, Grasiela Edith de Oliveira

AU - Nachum-Biala, Yaarit

AU - de Sousa, Keyla Carstens Marques

AU - da Silva, Thiago Merighi Vieira

AU - Campos, João Bosco Vilela

AU - Lemos, Wagner

AU - Moraes Barros-Battesti, Darci

AU - Machado, Rosangela Zacarias

AU - André, Marcos Rogério

N1 - Funding Information: The authors thank the Graduate Agricultural Microbiology Program (Universidade Estadual Paulista, UNESP, Campus Jaboticabal), the FAPESP (Foundation for Research Support of the State of São Paulo—Process 2018/02753‐0), FUNDECT (Foundation for Support to the Development of Education, Science and Technology of the State of Mato Grosso do Sul, Case 59/300.187/2016) and CNPq for the Productivity Grant granted to the MRA (CNPq Process No. 302420/2017‐7). The present study was carried out with the support of the Coordination for the Improvement of Higher Education (CAPES—Code 1). L.R.G. received a ‘Sandwich’ scholarship from the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior. Funding Information: The authors thank the Graduate Agricultural Microbiology Program (Universidade Estadual Paulista, UNESP, Campus Jaboticabal), the FAPESP (Foundation for Research Support of the State of São Paulo—Process 2018/02753-0), FUNDECT (Foundation for Support to the Development of Education, Science and Technology of the State of Mato Grosso do Sul, Case 59/300.187/2016) and CNPq for the Productivity Grant granted to the MRA (CNPq Process No. 302420/2017-7). The present study was carried out with the support of the Coordination for the Improvement of Higher Education (CAPES—Code 1). L.R.G. received a ‘Sandwich’ scholarship from the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior. Publisher Copyright: © 2020 Blackwell Verlag GmbH

PY - 2020/9/1

Y1 - 2020/9/1

N2 - Currently, five Bartonella species and an expanding number of Candidatus Bartonella species have globally been reported in ruminants. Likewise, different Bartonella genotypes were identified. However, studies relating to ruminant-associated Bartonella in Brazil are scarce. The current study aimed to assess the prevalence and genetic diversity of Bartonella in cattle, buffaloes and associated ectoparasites in Brazil. For this purpose, EDTA-blood samples from 75 cattle and 101 buffaloes were sampled. Additionally, 128 Rhipicephalus microplus and one Amblyomma sculptum ticks collected from cattle, and 197 R. microplus, one A. sculptum and 170 lice (Haematopinus tuberculatus) collected from buffaloes were included. Bartonella DNA was initially screened through an HRM real-time PCR assay targeting the 16S–23S internal transcribed spacer (ITS), and the positive samples were submitted to an additional HRM assay targeting the ssrA gene. The HRM-positive amplicons were sequenced, and the nucleotide identity was assessed by BLASTn. Bartonella spp.-positive DNA samples were analysed by conventional PCR assays targeting the gltA and rpoB genes, and then, the samples were cloned. Finally, the phylogenetic positioning and the genetic diversity of clones were assessed. Overall, 21 of 75 (28%) cattle blood samples and 13 of 126 (10.3%) associated ticks were positive for Bartonella bovis. Out of 101 buffaloes, 95 lice and 188 tick DNA samples, one (1%) buffalo and four (4.2%) lice were positive for Bartonella spp. Conversely, none of the ticks obtained from buffaloes were positive for Bartonella. The Bartonella sequences from buffaloes showed identity ranging from 100% (ITS and gltA) to 94% (ssrA) with B. bovis. In contrast, the Bartonella DNA sequences from lice were identical (100%) to uncultured Bartonella sp. detected in cattle tail louse (Haematopinus quadripertusus) from Israel in all amplified genes. The present study demonstrates the prevalence of new B. bovis genotypes and a cattle lice-associated Bartonella species in large ruminants and their ectoparasites from Brazil. These findings shed light on the distribution and genetic diversity of ruminant- and ectoparasite-related Bartonella in Brazil.

AB - Currently, five Bartonella species and an expanding number of Candidatus Bartonella species have globally been reported in ruminants. Likewise, different Bartonella genotypes were identified. However, studies relating to ruminant-associated Bartonella in Brazil are scarce. The current study aimed to assess the prevalence and genetic diversity of Bartonella in cattle, buffaloes and associated ectoparasites in Brazil. For this purpose, EDTA-blood samples from 75 cattle and 101 buffaloes were sampled. Additionally, 128 Rhipicephalus microplus and one Amblyomma sculptum ticks collected from cattle, and 197 R. microplus, one A. sculptum and 170 lice (Haematopinus tuberculatus) collected from buffaloes were included. Bartonella DNA was initially screened through an HRM real-time PCR assay targeting the 16S–23S internal transcribed spacer (ITS), and the positive samples were submitted to an additional HRM assay targeting the ssrA gene. The HRM-positive amplicons were sequenced, and the nucleotide identity was assessed by BLASTn. Bartonella spp.-positive DNA samples were analysed by conventional PCR assays targeting the gltA and rpoB genes, and then, the samples were cloned. Finally, the phylogenetic positioning and the genetic diversity of clones were assessed. Overall, 21 of 75 (28%) cattle blood samples and 13 of 126 (10.3%) associated ticks were positive for Bartonella bovis. Out of 101 buffaloes, 95 lice and 188 tick DNA samples, one (1%) buffalo and four (4.2%) lice were positive for Bartonella spp. Conversely, none of the ticks obtained from buffaloes were positive for Bartonella. The Bartonella sequences from buffaloes showed identity ranging from 100% (ITS and gltA) to 94% (ssrA) with B. bovis. In contrast, the Bartonella DNA sequences from lice were identical (100%) to uncultured Bartonella sp. detected in cattle tail louse (Haematopinus quadripertusus) from Israel in all amplified genes. The present study demonstrates the prevalence of new B. bovis genotypes and a cattle lice-associated Bartonella species in large ruminants and their ectoparasites from Brazil. These findings shed light on the distribution and genetic diversity of ruminant- and ectoparasite-related Bartonella in Brazil.

KW - Haematopinus tuberculatus

KW - Rhipicephalus microplus

KW - bartonellosis

KW - buffaloes

KW - cattle

KW - genetic diversity

UR - http://www.scopus.com/inward/record.url?scp=85081409309&partnerID=8YFLogxK

U2 - 10.1111/tbed.13517

DO - 10.1111/tbed.13517

M3 - Article

C2 - 32068964

AN - SCOPUS:85081409309

SN - 1865-1674

VL - 67

SP - 1888

EP - 1897

JO - Transboundary and Emerging Diseases

JF - Transboundary and Emerging Diseases

IS - 5

ER -

Molecular detection and genetic diversity of Bartonella species in large ruminants and associated ectoparasites from the Brazilian Cerrado

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Keywords

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