Bovine babesiosis: Anti-erythrocyte antibodies purification from the sera of naturally infected cattle

Goes, T.S.; Goes, Vinícius S.; Ribeiro, Múcio Flávio Barbosa; Gontijo, C.M.

doi:10.1016/j.vetimm.2006.12.011

Cited by 17 publications

(16 citation statements)

References 12 publications

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“…may cause direct damage on some erythrocytes, immune-mediated injury of parasite may be more important in the pathogenesis of anemia [7]. Yet, the increase in erythrophagocytosis by activated macrophages [50, 51] and the production of anti-erythrocyte antibodies [8] may also contribute to the development of anemia.…”

Section: Discussionmentioning

confidence: 99%

“…Because of its significant economic impact on meat and milk production and on livestock management, it is currently considered as the most important endemic parasitic disease affecting cattle in Egypt [5]. Both B. bigemina and B. bovis are capable of causing acute disease resulting in life-threatening hemolytic anemia as well as other severe clinical manifestations [7, 8]. Often, especially in bovines younger than a year old, acute disease resolves into persistent disease and the animals become carriers of the parasites, thus ensuring transmission by competent ticks in zoonotic areas.…”

Section: Introductionmentioning

confidence: 99%

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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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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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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“…This was a modified method as described previously [ 28 ]. The RBC white ghost membrane was used as antigen at a protein concentration of 2 μg in 100 μl of coating buffer (pH 9.6) per well to coat polystyrene plates (Lot number 091611, Nunc, Copenhagen, Denmark) at 4°C overnight.…”

Section: Methodsmentioning

confidence: 99%

Rate of red blood cell destruction varies in different strains of mice infected with Plasmodium berghei-ANKA after chronic exposure

Helegbe

Huy

Yanagi

et al. 2009

Malar J

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BackgroundSevere malaria anaemia in the semi-immune individuals in the holo-endemic area has been observed to occur at low parasite density with individual variation in the responses. Thus the following has been thought to be involved: auto-immune-mediated mechanisms of uninfected red blood cell destruction, and host genetic factors to explain the differences in individual responses under the same malaria transmission. In this study, the extent of red blood cell (RBC) destruction in different strains of semi-immune mice model at relatively low parasitaemia was studied.MethodologyTo generate semi-immunity, four strains of mice were taken through several cycles of infection and treatment. By means of immunofluorescent assay and ELISA, sera were screened for anti-erythrocyte auto-antibodies, and their relationship with haematological parameters and parasitaemia in the strains of semi-immune mice was investigated.ResultsUpon challenge with Plasmodium berghei ANKA after generating semi-immune status, different mean percentage haemoglobin (Hb) drop was observed in the mice strains (Balb/c = 47.1%; NZW = 30.05%; C57BL/6 = 28.44%; CBA = 25.1%), which occurred on different days for each strain (for Balb/c, mean period = 13.6 days; for C57BL/6, NZW, and CBA mean period = 10.6, 10.8, 10.9 days respectively). Binding of antibody to white ghost RBCs was observed in sera of the four strains of semi-immune mice by immunofluorescence. Mean percentage Hb drop per parasitaemia was highest in Balb/c (73.6), followed by C57BL/6 (8.6), CBA (6.9) and NZW (4.0), p = 0.0005. Consequently, auto-antibodies level to ghost RBC were correlated with degree of anaemia and were highest in Balb/c, when compared with the other strains, p < 0.001.ConclusionThe results presented in this study seem to indicate that anti-RBC auto-antibodies may be involved in the destruction of uninfected RBC in semi-immune mice at relatively low parasite burden. Host genetic factors may also influence the outcome of auto-immune mediated destruction of RBC due to the variation in Hb loss per % parasitaemia and differences in antibody titer for each semi-immune mice strain. However, further studies at the molecular level ought to be carried out to confirm this.

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“…A proporção superior de resultados falsos positivos no ELISA com CI em relação aos antígenos recombinantes provavelmente ocorreu devido a reações entre resíduos protéicos de eritrócitos, provenientes do processo de produção do antígeno, com anticorpos anti-eritrocíticos. Apesar do desenvolvimento desses anticorpos (anti-eritrocíti-cos) serem um processo patológico já bem documentado para alguns hemoprotozoários como Babesia gibsoni em cães (Adachi et al 1992), Babesia rodhaini e Plasmodium berghei em camundongos (Kanbara et al 1988), em bovinos apenas recentemente foi confirmado com Babesia bigemina (Góes et al 2007). Além disso, Madruga et al (2000) Outro ponto importante nesse trabalho foi a padronização de um ELISA utilizando como antígeno um coquetel de proteínas recombinantes.…”

Section: Resultsunclassified

“…Uma vez que os protocolos de infecção experimental por A. marginale geralmente utilizam inóculos contendo um grande número de riquétsias (10 10 eritrócitos infectados) (Palmer et al 1988, Gale et al 1996, Hope et al 2003, o que leva o animal a desenvolver altos títulos de anticorpos, os quais podem ser detectados por testes de menor sensibilidade. Por outro lado, os animais mantidos em isolamento ficam impossibilitados de contrair outros patógenos sanguíneos, como Babesia bovis, B. bigemina, Anaplasma phagocytophilum e A. bovis, que poderiam induzir à formação de anticorpos, os quais poderiam reagir cruzadamente com os antígenos de A. marginale, produzindo consequentemente resultados falsos positivos (Silva et al 2006, Santos & Carvalho 2006, Góes et al 2007.…”

Section: Methodsunclassified

Comparação entre diversos antígenos para o diagnóstico de Anaplasma marginale por ELISA

et al. 2010

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Anaplasmose bovina é uma doença com grande importância nas regiões tropicais e subtropicais do mundo por determinar perdas econômicas devido à mortalidade e redução da produtividade. É causada por Anaplasma marginale, uma riquétsia intraeritrocítica obrigatória cujo controle requer, além de uma vacina eficiente, uma acurada identificação de bovinos cronicamente infectados. Apesar de existirem atualmente diversos métodos de diagnóstico dessa riquétsia, os métodos sorológicos, em particular o ensaio de imunoadsorção enzimática-ELISAs, são os mais utilizados devido à sua versatilidade e praticidade. No entanto, devido ao grande número de antígenos disponíveis, atualmente torna-se necessária uma avaliação para definir quais antígenos apresentam um melhor desempenho no diagnóstico da anaplasmose. Soros de bovinos positivos e negativos para A. marginale por PCR, e soros de animais provenientes do Brasil e Costa Rica, foram testados em ELISAs baseados em MSP1a, MSP2 e MSP5 recombinantes, um pool das três proteínas recombinantes, e antígeno de lisado de corpúsculos iniciais da riquétsia (CI). Utilizando soro de bovinos positivos para A. marginale por PCR, uma maior sensibilidade foi observada no ELISA CI. No entanto, uma maior especificidade, com soro de bovinos negativos a PCR, foi observada com os ELISAs recombinantes. O porcentual de bovinos positivos do Brasil e Costa Rica foi maior com ELISA CI. Razões para essas diferenças são discutidas.

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Bovine babesiosis: Anti-erythrocyte antibodies purification from the sera of naturally infected cattle

Cited by 17 publications

References 12 publications

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

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

Rate of red blood cell destruction varies in different strains of mice infected with Plasmodium berghei-ANKA after chronic exposure

Comparação entre diversos antígenos para o diagnóstico de Anaplasma marginale por ELISA

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