Serologic immunoreactivity to Neospora caninum antigens in dogs determined by indirect immunofluorescence, western blotting and dot-ELISA

2005 ◽  
Vol 130 (1-2) ◽  
pp. 73-79 ◽  
Author(s):  
A.M. Pinheiro ◽  
M.F. Costa ◽  
B. Paule ◽  
V. Vale ◽  
M. Ribeiro ◽  
...  
Keyword(s):  
Western Blotting ◽  
Indirect Immunofluorescence ◽  
Neospora Caninum ◽  
Dot Elisa

scholarly journals Seroprevalence and risk factors associated with infection by Neospora caninum of dairy cattle in the state of Alagoas, Brazil

2012 ◽  
Vol 32 (10) ◽  
pp. 1009-1013 ◽  
Author(s):  
Maria E. Sousa ◽  
J.N. Porto Wagnner ◽  
Pedro P.F. Albuquerque ◽  
Orestes L. Souza Neto ◽  
Eduardo B. Faria ◽  
...  
Keyword(s):  
Risk Factors ◽  
Dairy Cattle ◽  
Indirect Immunofluorescence ◽  
Neospora Caninum ◽  
Infection Risk ◽  
Control Measures ◽  
Risk Indicators ◽  
Blood Samples ◽  
Factors Associated ◽  
Serological Investigation

The objective of this study was to investigate the prevalence of anti-Neospora caninum antibodies in cattle from milk producing farms of the microregion of Batalha, state of Alagoas, Brazil, as well as to identify the risk factors associated with the infection. Blood samples were collected from 1,004 cattle of 17 farms for the serological investigation regarding the presence of anti-N. caninum antibodies by the Indirect Immunofluorescence Reaction Technique (IMRT). From the total amount of samples analyzed, 77/1,004 (7.67%) were positive and 927/1,004 (92.33%) were negative. The logistical regression identified that cattle from farms without consortium breeding have an infection risk 6.33 (p<0.001; C.I. 2.89-13.10) times higher than cattle from farms with that type of breeding. Cattle from farms where the aborted fetuses are not adequately buried have an infection risk 3.04 (p<0.001; C.I. 1.64-5.63) times higher than cattle from farms with adequate destination of these fetuses. Infection by N. caninum occurs in cattle of the investigated region. The factors identified in our study can be used as risk indicators, so that control measures could be implemented to avoid infection by N. caninum in the herds of this region.

scholarly journals Desenvolvimento e padronização do Dot-ELISA usando peptídeos recombinantes para o diagnóstico sorológico de Neospora caninum

2014 ◽  
Vol 34 (8) ◽  
pp. 723-727
Author(s):  
Rafael D. Blanco ◽  
Cintia F. Fidelis ◽  
Leandro S. Araujo ◽  
Adriana M. Henao ◽  
Jose A. Cardona ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Neospora Caninum ◽  
E 92 ◽  
Dot Elisa

A neosporose é reconhecida como uma das maiores causas de aborto e perdas neonatais em bovinos de leite e corte em todo o mundo. Nos últimos anos esta doença tem atraído o interesse de pesquisadores com foco na epidemiologia e métodos eficazes de diagnóstico desta doença. No presente estudo objetivou-se desenvolver e padronizar um teste Dot-ELISA para o diagnóstico sorológico de Neospora caninum com um peptídeo recombinate como antígeno, visando o desenvolvimento de um kit para diagnóstico a campo. O peptídeo recombinante (rNcGRA1) foi desenhado com base na metodologia de genética reversa de epítopos antigênicos originados de uma proteína de grânulos densos de N. caninum, e sintetizado pela GenScript (USA). Produzido mediante o processo fermentativo em leveduras Pichia pastoris KM71. Para a padronização do Dot-ELISA, membranas de nitrocelulose de 0.22µm foram sensibilizadas com 1µL do antígeno e posteriormente os soros foram diluídos em solução de lavagem e incubados durante 1 hora. A revelação foi feita mediante a adição de Proteína G marcada com peroxidase por 30 minutos, seguido da solução reveladora a base de 3,3’-Diaminobenzidine (DAB). Logo após a padronização foram testados 44 soros bovinos diagnosticados por imunofluorescência indireta (RIFI), obtendo-se uma concordância nos resultados do teste de 95,5% e uma sensibilidade e especificidade de 100% e 92% respectivamente. Quanto ao Kit para diagnóstico a campo na Plataforma Tecnológica RapidFlow-Through Miriad®, o peptídeo rNcGRA1 apresentou marcações visíveis ao reagir com os soros positivos, e não apresentou marcações usando os soros negativos. Este estudo é o primeiro a utilizar peptídeos recombinantes e mostrar-se eficiente para o diagnóstico sorológico de bovinos naturalmente infetados por N. caninum.

scholarly journals Prevalence of Neospora caninum antibodies in sheep flocks of Uberlândia county, MG

2010 ◽  
Vol 19 (3) ◽  
pp. 148-151 ◽  
Author(s):  
Sandra Renata Sampaio Salaberry ◽  
Liria Hiromi Okuda ◽  
Alessandra Figueiredo de Castro Nassar ◽  
Jacqueline Ribeiro de Castro ◽  
Anna Monteiro Correia Lima-Ribeiro
Keyword(s):  
Risk Factors ◽  
Indirect Immunofluorescence ◽  
Neospora Caninum ◽  
Immunofluorescence Assay ◽  
Indirect Immunofluorescence Assay ◽  
Parasitic Disease ◽  
Blood Samples ◽  
Sheep Blood ◽  
Wild Canids ◽  
Significant Difference

Neosporosis is a parasitic disease that occurs in sheep and is associated with reproductive failure. The aim of this study was to verify seroprevalence of Neospora caninum antibodies in 12 sheep flocks in Uberlândia county, Minas Gerais, Brazil, using Indirect Immunofluorescence Assay (IFA). A total of 334 sheep blood samples were analyzed and an epidemiologic questionnaire was applied for each farm in order to correlate with risk factors of neosporosis: gender, age, breed, abortion problems, contact with cattle and dogs, and presence of wild canids. The prevalence of Neospora caninum antibodies in sheep was 8.1%, with titers ranging from 50 to 3200 and 83.3% of farms detected at least one ovine reactive. There was statistically significant difference in flocks with abortion problems associated with the presence of Neospora caninum antibodies (p < 0.05).

Platelets contain a 210K microtubule-associated protein related to a similar protein in HeLa cells

1988 ◽  
Vol 90 (2) ◽  
pp. 317-324
Author(s):  
F. Tablin ◽  
M.J. Reeber ◽  
V.T. Nachmias
Keyword(s):  
Western Blotting ◽  
Indirect Immunofluorescence ◽  
Polyclonal Antibody ◽  
Hela Cells ◽  
Blood Platelets ◽  
The Other ◽  
Similar Protein ◽  
Other Hand ◽  
Protein Map ◽  
Tubulin Immunofluorescence

We have demonstrated the presence of a 210K (K = 10(3) Mr) microtubule-associated protein (MAP) in blood platelets and have studied its relationship to tubulin and to the cytoskeleton, using a well-characterized polyclonal antibody for the analysis. When platelet lysates were enriched for tubulin by an assembly cycle at 37 degrees C, the 210K MAP was also enriched, as detected by Western blotting, while the antigen was not detected in pellets from cold-treated samples that lacked stabilized tubulin. Immunofluorescence of resting platelets showed that the 210K antigen colocalized with the microtubule coil in ring-like structures. On the other hand, in preparations of platelet cytoskeletons, the 210K antigen was present in samples from platelets in which the coil was disassembled (cold-treated without taxol pretreatment) as well as from platelets in which the coil was preserved (at 37 degrees C without taxol, or 4 degrees C with taxol pretreatment). In chilled platelets with disassembled microtubule coils, indirect immunofluorescence using antibodies to 210K or tubulin gave a diffuse signal throughout the platelet cytoplasm. However, immunofluorescence of the 210K antigen in both resting and cold-treated platelets displayed discrete or patchy staining as compared to the continuous staining with antitubulin. We conclude that 210K MAP is present in platelets, that it copurifies with tubulin and that it is localized along the microtubule coil. Our results also suggest that the 210K MAP may interact with some other element(s) of the cytoskeleton, and hence that it might serve as a linking protein.

scholarly journals TrkAIII Promotes Microtubule Nucleation and Assembly at the Centrosome in SH-SY5Y Neuroblastoma Cells, Contributing to an Undifferentiated Anaplastic Phenotype

2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
Antonietta R. Farina ◽  
Natalia Di Ianni ◽  
Lucia Cappabianca ◽  
Pierdomenico Ruggeri ◽  
Marzia Ragone ◽  
...  
Keyword(s):  
Cell Lines ◽  
Western Blotting ◽  
Indirect Immunofluorescence ◽  
Cell Morphology ◽  
Splice Variant ◽  
Neuroblastoma Cells ◽  
Advanced Stage ◽  
Microtubule Nucleation ◽  
Oncogenic Activity

The alternative TrkAIII splice variant is expressed by advanced stage human neuroblastomas (NBs) and exhibits oncogenic activity in NB models. In the present study, employing stable transfected cell lines and assays of indirect immunofluorescence, immunoprecipitation, Western blotting, microtubule regrowth, tubulin kinase, and tubulin polymerisation, we report that TrkAIII bindsα-tubulin and promotes MT nucleation and assembly at the centrosome. This effect depends upon spontaneous TrkAIII activity, TrkAIII localisation to the centrosome and pericentrosomal area, and the capacity of TrkAIII to bind, phosphorylate, and polymerise tubulin. We propose that this novel role for TrkAIII contributes to MT involvement in the promotion and maintenance of an undifferentiated anaplastic NB cell morphology by restricting and augmenting MT nucleation and assembly at the centrosomal MTOC.

scholarly journals Risk factors and anti-Toxoplasma gondii and Neospora caninum antibody occurrence in dogs in João Pessoa, Paraíba state, Northeastern Brazil

2018 ◽  
Vol 27 (2) ◽  
pp. 242-247 ◽  
Author(s):  
Arthur Willian de Lima Brasil ◽  
Roberta Nunes Parentoni ◽  
José Givanildo da Silva ◽  
Carolina de Sousa Américo Batista Santos ◽  
Rinaldo Aparecido Mota ◽  
...  
Keyword(s):  
Risk Factors ◽  
Toxoplasma Gondii ◽  
Indirect Immunofluorescence ◽  
Neospora Caninum ◽  
Northeastern Brazil ◽  
Blood Samples ◽  
Two Agents ◽  
Forested Areas ◽  
Caninum Antibody ◽  
The City

Abstract The aim of this study was to determine the occurrence of infections due to Toxoplasma gondii and Neospora caninum and corresponding risk factors among dogs attended at veterinary clinics in the city of João Pessoa, Paraíba, northeastern Brazil. Blood samples were collected from 384 dogs that were attended at 34 veterinary clinics between April 2015 and May 2016. These two agents were diagnosed through the indirect immunofluorescence reaction (IFAT). Among the 384 animals evaluated, 37 (9.6%) were positive for T. gondii, with titers ranging from 16 to 512. Six dogs (1.6%) were positive for N. caninum, with titers of 50 to 200. Access to the streets (OR = 4.60; 95% CI = 1.74-12.20) and environments close to forested areas (OR = 2.79; 95% CI = 1.32-5.93) were found to be risk factors for T. gondii infection. The dogs attended at veterinary clinics in João Pessoa are exposed to infections caused by T. gondii and N. caninum. Dog owners should avoid having access to the street or contact with forest environments.

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