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Previous issue date: 2010-02-10 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico, CNPq, Brasil. / The geographic distribution of bovine borreliosis is determined by the dispersion of its vector.
Borrelia theileri is the predominant species in cattle, and B. coriaceae and B. burgdorferi also
been reported causing clinical disease. B. theileri cause mild disease in cattle, and still is
important for its potential to be confused with the spirochete of Lyme disease, B. burgdorferi,
and agents of epizootic bovine abortion, B. coriaceae. In Brazil, as well as in other South
American countries, the agent of this disease has not been isolated further confusing the
diagnosis. The objective of this study was to identify genotypically Borrelia sp that affects
cattle in Brazil. DNA extraction, was performed from blood and ticks of cattle with positive
serology by indirect ELISA with crude antigen of Borrelia burgdorferi. Primers were
designed for genes of Borrelia burgdorferi and B. theileri groups: 16S, flaA, flaB, GroEL,
hbb, recA, 5s-23s, p66, rrs, rpoB and glpq. After the PCR reaction, only the primers amplified
rrs and rpoB sequences. The predictive amino acid sequence of RRS3 revealed 99%
homology with B. hermsii and B. duttonii and predictive amino acid sequence of RPOB
showed 67% homology with B. duttonii and B. recurrentis. This suggests that the species of
Borrelia sp present in Brazil is not owned by group B. burgdorferi.
Little is known regarding the genetic variability of genes that encode membrane proteins of
Brazilian isolates of A. marginale. The products of these genes constitute an important tool, as
there may be significant antigen polymorphism, which may damage cross-protection between
isolates and the chances of identifying candidate immunogens. The aim of the present study
was to determine the degree of conservation of sequences of these genes in a Brazilian isolate
of A. marginale comparing with Saint Maries and Florida isolates. For this, primers were
designed to amplify the genes omp1, omp4, omp5, omp7, omp8, omp10, omp14, omp15, sodb,
opag1, opag3, virb3, am097 (VirB9-1), am956 (PepA), am254 (ef-tu), am854 by PCR. The
genes were then sequenced by Sanger method and the predicted amino acid sequences aligned
and homology analyzed by the program CLUSTAL W. With the exception of OMP 7 all
proteins (OMP1, OMP4, OMP5, OMP8, OMP10, OMP14, OMP15, SODB, OPAG1,
OPAG3, VIRB3, VIRB9-1, PepA, EF-Tu, AM854) exhibited homology greater than 92%
with other A. marginale isolates. However, only OMP1, OMP5, EF-Tu, VirB3, SODB,
VIRB9-1 e AM854 showed homology greater than 72% regarding to A. marginale centrale
which confers cross-protection against A. marginale. / A distribui??o geogr?fica da borreliose bovina ? determinada pela dispers?o do seu vetor.
Borrelia theileri ? a esp?cie predominante em bovinos, sendo que B. burgdorferi e B.
coriaceae tamb?m foram relatadas causando doen?a cl?nica. Portanto, B. theileri causa doen?a
leve em bovinos, e ainda ? importante pelo seu potencial em ser confundido com a
espiroqueta da Doen?a de Lyme, B. burgdorferi, e com agentes do Aborto Epizo?tico bovino,
B. coriaceae. No Brasil, assim como em outros pa?ses Sul americanos, o agente desta
enfermidade ainda n?o foi isolado prejudicando ainda mais o diagn?stico. O objetivo deste
trabalho foi ? identifica??o genot?pica da esp?cie de Borrelia sp que acomete bovinos no
Brasil. Foram utilizados para extra??o de DNA, o sangue e carrapatos de bovinos com
sorologia positiva ao ELISA indireto com ant?geno bruto para Borrelia burgdorferi. Foram
desenhados oligonucleot?deos iniciadores para genes dos grupos Borrelia burgdorferi e B.
theileri: 16S, flaA, flaB, groel, hbb, recA, 5s-23s, p66, rrs, rpob e glpq. Ap?s a rea??o de
PCR, somente os oligonucleot?deos iniciadores rrs e rpob amplificaram seq??ncias. A
seq??ncia preditiva de amino?cidos de RRS3 revelou homologia de 99% com B. hermsii e B.
duttonii e a seq??ncia preditiva de amino?cidos de RPOB demonstrou 67% de homologia com
B. duttonii e B. recurrentis. Isto sugere que a esp?cie de Borrelia presente no Brasil n?o seja
pertencente ao grupo de B. burgdorferi.
Pouco se sabe sobre a variabilidade gen?tica dos genes que codificam prote?nas de membrana
de isolados brasileiros de A. marginale. O produto destes genes constitui uma ferramenta
importante, pois pode haver polimorfismo antig?nico, que pode prejudicar a prote??o cruzada
entre os isolados e as chances de identifica??o de candidatos a imun?genos. O objetivo do
presente estudo foi determinar o grau de conserva??o das seq??ncias destes genes em um
isolado brasileiro de A. marginale frente aos isolados Saint Maries, Florida e A. marginale
centrale. Para tanto, oligonucleot?deos foram desenhados para amplificar os genes omp1,
omp4, omp5, omp7, omp8, omp10, omp14, omp15, sodb, opag1, opag3, virb3, am097 (VirB9-
1), am956 (PepA), am254 (ef-tu), am854 por PCR. Os genes foram ent?o seq?enciados pelo
m?todo de Sanger e as seq??ncias preditas de amino?cidos alinhadas e a homologia analisada
atrav?s do programa CLUSTAL W. Com exce??o de OMP 7 todas as demais (OMP1, OMP4,
OMP5, OMP8, OMP10, OMP14, OMP15, SODB, OPAG1, OPAG3, VIRB3, VIRB9-1,
PepA, EF-Tu, AM854) apresentaram n?veis de homologia de 92 a 100% entre os isolados de
A. marginale. Destas, apenas OMP1, OMP5, EF-Tu, VirB3, SODB, VIRB9-1 e AM854
apresentaram homologia superior a 72% em rela??o a A. marginale centrale, o qual confere
prote??o cruzada contra A. marginale.
| Identifer | oai:union.ndltd.org:IBICT/oai:localhost:jspui/1860 |
| Date | 10 February 2010 |
| Creators | Daniel da Silva, Guedes Junior |
| Contributors | Fonseca, Adivaldo Henrique da, Ara?jo, Fl?bio Ribeiro de, Fragoso, Stenio Perdig?o, Massard, Carlos Luiz, Botteon, Paulo de Tarso Landgarff, Barreiras, Jairo Dias |
| Publisher | Universidade Federal Rural do Rio de Janeiro, Programa de P?s-Gradua??o em Ci?ncias Veterin?rias, UFRRJ, Brasil, Instituto de Veterin?ria |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/doctoralThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da UFRRJ, instname:Universidade Federal Rural do Rio de Janeiro, instacron:UFRRJ |
| Rights | info:eu-repo/semantics/openAccess |
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