Semi-quantitative evaluation of Babesia bovis and B. bigemina infection levels estimated by HRM analysis

Author(s)

Giglioti, Rodrigo ^[1] ; Okino, Cintia Hiromi ^[2] ; Azevedo, Bianca Taina ^[1] ; Rodrigues Wedy, Bruna Costa ^[1] ; Gutmanis, Gunta ^[1] ; Verissimo, Cecilia Jose ^[1] ; Katiki, Luciana Morita ^[1] ; Vercesi Filho, Anibal Eugenio ^[1] ; de Oliveira, Henrique Nunes ^[3] ; de Sena Oliveira, Marcia Cristina ^[2]

Affiliation

^[1] Inst Zootecnia, Rua Heitor Penteado 56, BR-13380011 Nova Odessa, SP - Brazil

^[2] Embrapa Pecuaria Sudeste, Sao Carlos, SP - Brazil

^[3] Univ Estadual Julio de Mesquita Filho, Unesp Jaboticabal, Sao Paulo - Brazil

• Document type: Journal article
• Source: TICKS AND TICK-BORNE DISEASES; v. 12, n. 5 SEP 2021.
• Web of Science Citations: 0

Abstract

Bovine babesiosis is economically the most important arthropod-borne disease of cattle worldwide. The most significant damage caused by bovine babesiosis is attributed to Babesia bovis due to its higher pathogenicity. This study aimed to develop a real-time PCR method followed by HRM (high-resolution melting) analysis for the simultaneous detection of B. bovis and B. bigemina, enabling a semi-quantitative analysis of Babesia levels using a single-tube reaction. The HRM was compared with real-time PCR using species-specific hydrolysis probes. The HRM analysis allowed to differentiate both Babesia species and was sensitive in the detection and differentiation of 10% for each Babesia species in the sample. Our results suggest the use of this method to estimate the prevalence of infections by B. bovis or B. bigemina as an alternative to the methods of absolute quantification by real-time PCR since it neither requires precise estimates of the number of DNA loads nor the construction of calibration curves. The simultaneous detection of the two Babesia species can be used to characterise the infection levels in cattle populations from different geographical regions, allowing a better control of these diseases. (AU)

• FAPESP's process: 19/22675-6 - Transcriptome analysis of resistance and/or susceptibility in beef cattle of different genetic groups to Babesia bovis infections
• Grantee: Rodrigo Giglioti
• Support Opportunities: Regular Research Grants
• FAPESP's process: 16/07216-7 - Genomic study and immunological characterization of cattle resistant to bovine babesiosis and analysis of the genetic diversity of Babesia bovis and Babesia bigemina
• Grantee: Henrique Nunes de Oliveira
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 18/19452-2 - Multi user equipment approved in grant 2017/50.339-5: nanoespectrofotômetro BioDrop
• Grantee: Renata Helena Branco Arnandes
• Support Opportunities: Multi-user Equipment Program