Use of an immunodominant p17 antigenic fraction of Neospora caninum in detection of antibody response in cattle

García, Gema Álvarez; Pérez, Inmaculada López; Innes, Elisabeth A.; Fernández, Esther Collantes; García, Aurora Fernández; Bautista, Mercedes Gómez; Ortega‐Mora, Luis Miguel

doi:10.1590/s0074-02762006000500008

Cited by 7 publications

(4 citation statements)

References 17 publications

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“…The use of sNcAg as effective vaccine antigen is supported by several studies performed by our group and others (Alvarez Garcia et al, 2006;Baszler et al, 2000;Mansilla et al, 2012;Moore et al, 2011). Soluble antigens of N. caninum tachyzoites can activate CD4 + T-cells (Rocchi et al, 2011), which are thought to be important in protecting cattle against this parasite's infection (Rocchi et al, 2011;Staska et al, 2003).…”

Section: Discussionmentioning

confidence: 89%

“…However, the use of live vaccines can have some disadvantages including reversion to high virulence creating sources of persistent infections, risk of transmission of prion diseases and high costs for industrial production. Although there was evidence of efficacy using a commercial vaccine based on a crude lysate of tachyzoites (Romero et al, 2004) a recent clinical trial concluded that vaccination after conception prevented 61% abortions in one of five herds and that vaccination may have increased the risk of early embryonic death (Weston et al, 2012), Fractions of N. caninum soluble native antigens (sNcAg) constitute a promising candidate antigen for vaccine development (Alvarez Garcia et al, 2006;Baszler et al, 2000;Mansilla et al, 2012;Moore et al, 2011;Otsuki et al, 2013), as it does not require inactivation treatments that could modify and ruin antigenic determinants. Proteins present in the soluble-fraction have been identified as stimulators of CD4 + -T cell responses (Rocchi et al, 2011;Tuo et al, 2005) and dendritic cell maturation, promoting the acquired immunity, which could not be achieved using inactivated parasites (Mansilla et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

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Dose-dependent immunogenicity of a soluble Neospora caninum tachyzoite-extract vaccine formulated with a soy lecithin/β-glucan adjuvant in cattle

Mansilla¹,

Czepluch²,

Malacari³

et al. 2013

Veterinary Parasitology

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Section: Discussionmentioning

confidence: 89%

Section: Introductionmentioning

confidence: 99%

Dose-dependent immunogenicity of a soluble Neospora caninum tachyzoite-extract vaccine formulated with a soy lecithin/β-glucan adjuvant in cattle

Mansilla¹,

Czepluch²,

Malacari³

et al. 2013

Veterinary Parasitology

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“…In a continued effort to identify useful N. caninum antigens for the immunodiagnosis of neosporosis, we report here the molecular characterization of the 17-19 kDa antigenic protein fraction, p17, previously identified as the major IDA in onedimensional PAGE by sera from bovines naturally infected with Neospora (Alvarez- Garcia et al 2002;Alvarez-Garcia et al 2006). Further characterization of p17 was accomplished by 2 complementary approaches.…”

Section: Discussionmentioning

confidence: 99%

“…(Alvarez-Garcia et al 2002;Alvarez-Garcia et al 2006). Further characterization of p17 was accomplished by 2 complementary approaches.…”

mentioning

confidence: 99%

The NcGRA7gene encodes the immunodominant 17 kDa antigen ofNeospora caninum

et al. 2006

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A Neospora caninum 17-19 kDa antigenic protein fraction (p17) in one-dimensional polyacrylamide gel electrophoresis (SDS-PAGE) is the immunodominant antigen recognized by sera from bovines naturally infected by N. caninum. To identify the proteins making up the p17 fraction, we screened a new N. caninum tachyzoite cDNA library with an affinity-purified antibody against p17 (APA17). We isolated several cDNA clones with 100% sequence identity to the NcGRA7 gene. This previously described gene encodes a dense granule protein with an apparent molecular mass of 33 kDa. A second line of evidence emerged through a combined proteomic approach associating two-dimensional PAGE (2D-PAGE) to Western blotting and to mass spectrometry to characterize the p17 fraction. Two acidic immunodominant but minority protein spots were recognized by APA17 and by bovine sera. These antigens of 17 and 33 kDa are respectively composed of 4 and 2 isoforms. Furthermore, p17 isolation by 2D-PAGE and peptide sequencing by tandem mass spectrometry yielded a partial sequence of 17 amino acids, which allowed the putative amino terminal region of the NcGRA7 protein to be identified unambiguously. The NcGRA7 protein, without the putative signal peptide at the NH2-terminus, was cloned and expressed in Escherichia coli and when the purified recombinant protein (rNcGRA7) was analysed by SDS-PAGE and mass spectrometry, 2 bands of 24 and 33 kDa were resolved and identified as NcGRA7. These results demonstrate that the immunodominant 17 kDa antigen of N. caninum is encoded by the NcGRA7 gene.

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Seroprevalence and associated risk factors for Neospora caninum infection in dairy cattle in South Africa

Tagwireyi,

Thompson,

Garcia

et al. 2024

Parasitol Res

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Bovine neosporosis is a widespread parasitic disease associated with significant economic losses. Its effects on the reproductive performance of cows have resulted in losses that run into the hundreds of millions of US dollars in dairy industries in various countries (Reichel et al., Int J Parasitol 43:133–142, 2013). Due to outdated and scant information on the occurrence of Neospora caninum infection in South Africa, the study aimed to determine the seroprevalence and risk factors associated with infection in dairy cattle in South Africa. A total of 1401 blood samples were randomly collected from cattle on 48 dairy farms in seven of the nine provinces in South Africa. A close-ended questionnaire was used in a cross-sectional study to obtain farm-level and animal-level data. Serological testing was done using a commercial IDvet Screen® Neospora caninum Indirect ELISA. An overall seroprevalence, adjusted for test sensitivity and specificity, of 2.3% (95% CI, 1.3–4.1) was detected and 48% (23/48) of sampled farms had at least one animal testing positive. The highest seroprevalence of N. caninum was in the KwaZulu-Natal province with 7.5% (95% CI, 3.8–14.3), and the lowest in Western Cape with 0.1% (95% CI, 0–1.2). The highest within-farm seroprevalence of 25% was detected on a farm in the North West Province. In a multivariable logistic regression model, the odds of N. caninum seropositivity were higher in Holstein–Friesian cattle when compared to other breeds. Good hygiene was identified as a protective factor. Cattle left out on pasture had increased odds of testing positive for N. caninum compared to those that were penned. The odds of testing seropositive for N. caninum was higher on farms that practised segregation of cattle into different age groups. The purchase of replacement animals was a significant risk factor, as open herds had increased odds of N. caninum seropositivity. Cattle on farms that did not have a specific calving location were more likely to be seropositive. This is the first such study in South Africa and shows that N. caninum is widely distributed in the country at a low seroprevalence, but it may be a cause of concern on certain farms.

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Use of an immunodominant p17 antigenic fraction of Neospora caninum in detection of antibody response in cattle

Cited by 7 publications

References 17 publications

Dose-dependent immunogenicity of a soluble Neospora caninum tachyzoite-extract vaccine formulated with a soy lecithin/β-glucan adjuvant in cattle

Dose-dependent immunogenicity of a soluble Neospora caninum tachyzoite-extract vaccine formulated with a soy lecithin/β-glucan adjuvant in cattle

The NcGRA7gene encodes the immunodominant 17 kDa antigen ofNeospora caninum

Seroprevalence and associated risk factors for Neospora caninum infection in dairy cattle in South Africa

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