Datos de investigación
Improved Serodiagnosis of Trypanosoma vivax Infections in Cattle Reveals High Infection Rates in the Livestock Regions of Argentina
Publicador:
Consejo Nacional de Investigaciones Científicas y Técnicas
Fecha de depósito:
06/05/2024
Fecha de recolección:
02/01/2021-05/01/2023
Clasificación temática:
Resumen
Bovine trypanosomosis, caused by Trypanosoma vivax, currently affects cattle and has a significant economic impact in sub-Saharan Africa and South America. The development of new diagnostic antigens is essential to improve and refine existing methods. Our study evaluated the efficacy of two recombinant antigens in detecting specific antibodies in cattle. These antigens are derivatives of an invariant surface glycoprotein (ISG) from T. vivax. A fraction of a previously described antigen (TvY486_0045500), designated TvISGAf, from an African strain was evaluated, and a new ISG antigen from an American isolate, TvISGAm, was identified. The two antigens were expressed as fusion proteins in Escherichia coli: TvISGAf was fused to the MBP-His-tag, and TvISGAm was obtained as a His-tag fused protein. An ELISA evaluation was conducted using these antigens on 149 positive and 63 negative bovine samples. The diagnostic performance was enhanced by the use of a combination of both antigens (referred to as TvISG-based ELISA), achieving a sensitivity of 89.6% and specificity of 93.8%. Following the validation of the TvISG-based ELISA, the seroprevalence of T. vivax infection in 892 field samples from cattle in the central region of Argentina was determined. The mean seroprevalence of T. vivax was 53%, with variation ranging from 21% to 69% among the six departments studied. These results support the use of the TvISG ELISA as a valuable serological tool for the detection and monitoring of T. vivax infection in cattle. Furthermore, we report for the first time the seroprevalence of T. vivax in Argentina, which highlights the widespread endemic nature of the disease in the region. In order to effectively manage the increasing spread of T. vivax in the vast livestock production areas of South America, it is essential to implement consistent surveillance programs and to adopt preventive strategies.
Métodos
resumir: Immunodetection experiments were adapted from a previously described method for T. cruzi (31). Briefly, polyclonal antibodies against TvISGAf and TvISGAm were obtained by immunizing mice with 10 µg of purified recombinant protein at two-week intervals, as previously described (32). T. vivax cells of the Y486 strain, obtained from mice infected for four days, were pelleted and washed twice for 15 minutes at room temperature with sterile PBS buffer and then fixed in 4% (v/v) formaldehyde. After washing, they were permeabilized and blocked for 30 minutes in a medium containing PBS with the addition of 0.1% (v/v) Triton X-100 and 3% (w/v) BSA. The primary antibodies used were either mouse anti-TvISAf sera (1/100) or mouse anti-TvISGAm sera (1/100), along with rabbit anti-TccPx polyclonal antibodies (1/100 dilution). Slide wells were washed three times with blocking solution (3x50 µl). Secondary antibodies diluted 1/100 in blocking buffer (20 µl) were added to the slide wells and incubated for 60 minutes. FITC-conjugated goat anti-rabbit or Alexa 680-conjugated goat anti-mouse antibodies were used as secondary antibodies. DAPI (20 µl/slide well), diluted in PBS (final concentration 0.5 µg/ml), was added to each slide well. Slides were examined with a confocal microscope (Leica).
Palabras clave:
T. vivax seroprevalence,
Dairy cattle,
Endemic disease,
Parasite spreading
Alcance geográfico
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Identificador del recurso
Colecciones
Datos de Investigación(CCT - SANTA FE)
Datos de Investigación de CTRO.CIENTIFICO TECNOL.CONICET - SANTA FE
Datos de Investigación de CTRO.CIENTIFICO TECNOL.CONICET - SANTA FE
Datos de Investigación(IAL)
Datos de Investigación de INSTITUTO DE AGROBIOTECNOLOGIA DEL LITORAL
Datos de Investigación de INSTITUTO DE AGROBIOTECNOLOGIA DEL LITORAL
Citación
Bontempi, Iván; Arias, Diego Gustavo; Marcipar, Iván Sergio; (2024): Improved Serodiagnosis of Trypanosoma vivax Infections in Cattle Reveals High Infection Rates in the Livestock Regions of Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. (dataset). http://hdl.handle.net/11336/234547
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