Virulence studies of Neospora caninum in goat trophoblast primary culture

Neospora caninum is a well-documented cause of abortion in cattle worldwide. The relevance of the abortifacient role of this parasite in small ruminants remains under investigation, especially in areas with high seroprevalence. The objective of this study was to evaluate the virulence (tachyzoite yi...

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Autores principales: Álvarez, Betiana Elizabeth, Rambeaud, Magdalena, Dellarupe, Andrea, Campero, Lucía María, Venturini, María Cecilia
Formato: Objeto de conferencia Resumen
Lenguaje:Inglés
Publicado: 2024
Materias:
Ciencias Veterinarias
Neospora caninum
cultivo celular
trofoblasto
invasión
inmunofluorescencia
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/176905
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-176905
record_format dspace
spelling I19-R120-10915-1769052025-02-25T20:28:53Z http://sedici.unlp.edu.ar/handle/10915/176905 Virulence studies of Neospora caninum in goat trophoblast primary culture Álvarez, Betiana Elizabeth Rambeaud, Magdalena Dellarupe, Andrea Campero, Lucía María Venturini, María Cecilia 2024-10-25 2024 2025-02-25T16:10:26Z en Ciencias Veterinarias Neospora caninum cultivo celular trofoblasto invasión inmunofluorescencia Neospora caninum is a well-documented cause of abortion in cattle worldwide. The relevance of the abortifacient role of this parasite in small ruminants remains under investigation, especially in areas with high seroprevalence. The objective of this study was to evaluate the virulence (tachyzoite yield [TY] and invasion rate [IR]) of a locally isolated N. caninum strain (NC-Argentina LP1) compared to a reference strain (NC-1) in goat trophoblast cells. Primary culture was obtained from cotyledons of a goat seronegative for N. caninum and Toxoplasma gondii, and negative by PCR, using the explant method. A total of 105placental cells/well (passage 4) were seeded into 24-well plates and grown to confluence (48 hours). For TY and IR studies, cultures were infected with 2x105tachyzoites/well and 104tachyzoites/well of each strain, respectively, in 3 independent assays. At 48 h post infection, TY was quantified by qPCR, and IR was evaluated by immunolabeling of parasitophorous vacuoles (PVs) with anti-N. caninum hyperimmune serum and its conjugate (Alexa Fluor 488, Life Technologies) and DAPI staining. Fifty fields per replicate were counted and PV size was measured. Regarding TY, significant differences were found between NC-Argentina LP1 and NC-1, with lower values for NC-Argentina LP1 compared to NC-1 (p<0.01). In addition, NC-Argentina LP1 showed a lower IR than the reference strain (p<0.0001). However, the size of the PVs was larger for NC-Argentina LP1 with respect to NC-1 (p<0.0001), which could be related to its lower capacity to reinvade. These results suggest that NC-Argentina LP1 can invade and replicate in primary cultures of goat trophoblast, although with lower virulence than the reference strain NC-1, and may aid in predicting in vivo behavior. Facultad de Ciencias Veterinarias Objeto de conferencia Resumen http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf 122-123
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Ciencias Veterinarias
Neospora caninum
cultivo celular
trofoblasto
invasión
inmunofluorescencia
spellingShingle Ciencias Veterinarias
Neospora caninum
cultivo celular
trofoblasto
invasión
inmunofluorescencia
Álvarez, Betiana Elizabeth
Rambeaud, Magdalena
Dellarupe, Andrea
Campero, Lucía María
Venturini, María Cecilia
Virulence studies of Neospora caninum in goat trophoblast primary culture
topic_facet Ciencias Veterinarias
Neospora caninum
cultivo celular
trofoblasto
invasión
inmunofluorescencia
description Neospora caninum is a well-documented cause of abortion in cattle worldwide. The relevance of the abortifacient role of this parasite in small ruminants remains under investigation, especially in areas with high seroprevalence. The objective of this study was to evaluate the virulence (tachyzoite yield [TY] and invasion rate [IR]) of a locally isolated N. caninum strain (NC-Argentina LP1) compared to a reference strain (NC-1) in goat trophoblast cells. Primary culture was obtained from cotyledons of a goat seronegative for N. caninum and Toxoplasma gondii, and negative by PCR, using the explant method. A total of 105placental cells/well (passage 4) were seeded into 24-well plates and grown to confluence (48 hours). For TY and IR studies, cultures were infected with 2x105tachyzoites/well and 104tachyzoites/well of each strain, respectively, in 3 independent assays. At 48 h post infection, TY was quantified by qPCR, and IR was evaluated by immunolabeling of parasitophorous vacuoles (PVs) with anti-N. caninum hyperimmune serum and its conjugate (Alexa Fluor 488, Life Technologies) and DAPI staining. Fifty fields per replicate were counted and PV size was measured. Regarding TY, significant differences were found between NC-Argentina LP1 and NC-1, with lower values for NC-Argentina LP1 compared to NC-1 (p<0.01). In addition, NC-Argentina LP1 showed a lower IR than the reference strain (p<0.0001). However, the size of the PVs was larger for NC-Argentina LP1 with respect to NC-1 (p<0.0001), which could be related to its lower capacity to reinvade. These results suggest that NC-Argentina LP1 can invade and replicate in primary cultures of goat trophoblast, although with lower virulence than the reference strain NC-1, and may aid in predicting in vivo behavior.
format Objeto de conferencia
Resumen
author Álvarez, Betiana Elizabeth
Rambeaud, Magdalena
Dellarupe, Andrea
Campero, Lucía María
Venturini, María Cecilia
author_facet Álvarez, Betiana Elizabeth
Rambeaud, Magdalena
Dellarupe, Andrea
Campero, Lucía María
Venturini, María Cecilia
author_sort Álvarez, Betiana Elizabeth
title Virulence studies of Neospora caninum in goat trophoblast primary culture
title_short Virulence studies of Neospora caninum in goat trophoblast primary culture
title_full Virulence studies of Neospora caninum in goat trophoblast primary culture
title_fullStr Virulence studies of Neospora caninum in goat trophoblast primary culture
title_full_unstemmed Virulence studies of Neospora caninum in goat trophoblast primary culture
title_sort virulence studies of neospora caninum in goat trophoblast primary culture
publishDate 2024
url http://sedici.unlp.edu.ar/handle/10915/176905
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AT dellarupeandrea virulencestudiesofneosporacaninumingoattrophoblastprimaryculture
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AT venturinimariacecilia virulencestudiesofneosporacaninumingoattrophoblastprimaryculture
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