Validation and Field Evaluation of a Competitive Enzyme-Linked Immunosorbent Assay for Diagnosis of Babesia bovis Infections in Argentina

Dominguez, Mariana Ribeiro; Echaide, Ignacio; Echaide, Susana Torioni de; Wilkowsky, Silvina Elizabeth; Zábal, Osvaldo; Mosqueda, Juan; Schnittger, Leonhard; Florín-Christensen, Mónica

doi:10.1128/cvi.00015-12

Cited by 17 publications

(12 citation statements)

References 28 publications

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“…It is important to note that bovines immunized with VAC-LC do not show the clinical signs observed in cattle inoculated with VAC after vaccination [19,20]; however, the underlying mechanism responsible for this protection is not known. The only evidence we have is the recorded level of total IgG anti-Babesia [21], which is higher in animals treated with VAC and L. casei than in bovines immunized with VAC [18,19]. To date, there are no published studies on the levels of IgG1 and IgG2 against Babesia in bovines immunized with VAC or VAC and L. casei.…”

Section: Introductioncontrasting

confidence: 47%

Co-immunization of cattle with a vaccine against babesiosis and Lactobacillus casei increases specific IgG1 levels to Babesia bovis and B. bigemina

Garfias

Lozano

Martı́nez

et al. 2015

Parasitology International

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Section: Introductioncontrasting

confidence: 47%

Co-immunization of cattle with a vaccine against babesiosis and Lactobacillus casei increases specific IgG1 levels to Babesia bovis and B. bigemina

Garfias

Lozano

Martı́nez

et al. 2015

Parasitology International

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“…In addition, cELISA has the potential to display higher specificity than an indirect ELISA [35]. In the current study, the cELISA assays detected a higher number of infected animals than the PCR assays; this could be explained by differences in the timing of the parasite presence and the antibody responses in the infected animals as well as the stage of infections [36, 37].…”

Section: Discussionmentioning

confidence: 86%

Serological and molecular diagnostic surveys combined with examining hematological profiles suggests increased levels of infection and hematological response of cattle to babesiosis infections compared to native buffaloes in Egypt

Mahmoud¹,

Kandil²,

Nasr³

et al. 2015

Parasites Vectors

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BackgroundBabesiosis threatens the development of the cattle and buffaloes industries in Egypt and improved control is needed. The main objectives of this study are surveying the presence of bovine babesiosis in distinct selected bovine and buffalo populations in Egypt using novel molecular and previously validated serological methods, while also comparing the occurrence of hematological alterations among Babesia infected cattle and buffalos.MethodsA total of 253 and 81 blood samples from apparently healthy cattle and buffaloes, respectively, were randomly collected from diverse locations in Egypt. All samples were tested for Babesia bovis and B. bigemina infection using blood film examination, competitive ELISA (cELISA) and PCR. Novel semi-nested and nested PCR assays for the detection of B. bovis and B. bigemina respectively, were developed and used to analyze DNA extracted from bovine and buffalo samples. Hematological profiles were studied using a hematological analyzer.ResultsBlood films examination revealed 13.8 % and 7.4 % Babesia infection rates in cattle and buffaloes, respectively. However, in cattle, the cELISA detected 32.8 %, 21.3 % and 10.7 % infection rates with B. bigemina, B. bovis and mixed infection, respectively. In addition, cELISA identified 22.2 %, 22.2 % and 6.2 % infection rates with B. bigemina, B. bovis and mixed infection, respectively in buffaloes. The semi-nested PCR assay showed that 15 % of the tested samples were positive for B. bovis in cattle, but just 3 % in buffaloes. Infections with B. bigemina were also found in cattle (32.4 %), but not in buffaloes upon nested PCR analysis. Sequencing analysis confirmed the identity of the PCR amplicons and showed that Egyptian genotypes of B. bigemina and B. bovis highly resemble sequences previously deposited in GenBank. Hemograms performed on the sampled animals revealed macrocytic hypochromic anemia associated with reduced platelet counts in infected cattle with babesiosis. In addition, marked increases in total leukocyte and granulocytic counts and decreases in lymphocytic counts were found in infected cattle. In contrast, no such hematological anomalies were found in presumably Babesia-infected buffaloes.ConclusionsFrequent occurrence of babesiosis among apparently healthy bovines in Egypt, suggests the need for appropriately designed prevalence studies in this country. Infected bovine, but not buffalo, populations often present hematological disorders compatible with intravascular hemolysis and thrombocytopenia.

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“…Immunofluorescence assays were performed as described [18]. Briefly, smears were prepared using a suspension of B. bovis (BboS2P pathogenic strain, Argentina)-parasitized erythrocytes (13% infection) that were cultured in vitro and diluted 1:3 in PBS containing 1% BSA.…”

Section: Methodsmentioning

confidence: 99%

“…As invasion of red blood cells is crucial to the survival of the parasite, blocking the antigens involved in this process may lead to protection. MSA-2c is highly conserved among B. bovis geographical isolates and therefore, together with its high immunogenicity during infection of cattle, it has been applied to the development of serological diagnostic methods [11, 13, 14, 17, 18]. …”

Section: Introductionmentioning

confidence: 99%

A recombinant multi-antigen vaccine formulation containing Babesia bovis merozoite surface antigens MSA-2a1, MSA-2b and MSA-2c elicits invasion-inhibitory antibodies and IFN-γ producing cells

et al. 2016

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Background Babesia bovis is a tick-transmitted protozoan hemoparasite and the causative agent of bovine babesiosis, a potential risk to more than 500 million cattle worldwide. The vaccines currently available are based on attenuated parasites, which are difficult to produce, and are only recommended for use in bovines under one year of age. When used in older animals, these vaccines may cause life-threatening clinical symptoms and eventually death. The development of a multi-subunit recombinant vaccine against B. bovis would be attractive from an economic standpoint and, most importantly, could be recommended for animals of any age. In the present study, recombinant ectodomains of MSA-2a1, MSA-2b and MSA-2c antigens were expressed in Pichia pastoris yeast as secreted soluble peptides.ResultsThe antigens were purified to homogeneity, and biochemically and immunologically characterized. A vaccine formulation was obtained by emulsifying a mixture of the three peptides with the adjuvant Montanide ISA 720, which elicited high IgG antibody titers against each of the above antigens. IgG antibodies generated against each MSA-antigen recognized merozoites and significantly inhibited the invasion of bovine erythrocytes. Cellular immune responses were also detected, which were characterized by splenic and lymph node CD4+ T cells producing IFN-γ and TNF-α upon stimulation with the antigens MSA-2a1 or MSA-2c.ConclusionsThese data strongly suggest the high protective potential of the presented formulation, and we propose that it could be tested in vaccination trials of bovines challenged with B. bovis.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1862-1) contains supplementary material, which is available to authorized users.

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Validation and Field Evaluation of a Competitive Enzyme-Linked Immunosorbent Assay for Diagnosis of Babesia bovis Infections in Argentina

Cited by 17 publications

References 28 publications

Co-immunization of cattle with a vaccine against babesiosis and Lactobacillus casei increases specific IgG1 levels to Babesia bovis and B. bigemina

Co-immunization of cattle with a vaccine against babesiosis and Lactobacillus casei increases specific IgG1 levels to Babesia bovis and B. bigemina

Serological and molecular diagnostic surveys combined with examining hematological profiles suggests increased levels of infection and hematological response of cattle to babesiosis infections compared to native buffaloes in Egypt

A recombinant multi-antigen vaccine formulation containing Babesia bovis merozoite surface antigens MSA-2a1, MSA-2b and MSA-2c elicits invasion-inhibitory antibodies and IFN-γ producing cells

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