Resultado da pesquisa (113)

Termo utilizado na pesquisa analysis

#101 - Clonal study of avian Escherichia coli strains by fliC conserved-DNA-sequence regions analysis, p.508-514

Abstract in English:

ABSTRACT.- Campos T.A., Nakazato G., Stehling E.G., Brocchi M. & Silveira W.D. 2008. Clonal study of avian Escherichia coli strains by fliC conserved-DNA-sequence regions analysis. Pesquisa Veterinária Brasileira 28(10):508-514. Departamento de Microbiologia e Imunologia, Instituto de Biologia, Cx. Postal 6109, Universidade Estadual de Campinas, Cidade Universitária Zeferino Vaz s/n, Barão Geraldo, Campinas, SP 3081-862, Brazil. *Corresponding author: wds@unicamp.br The clonal relationship among avian Escherichia coli strains and their genetic proximity with human pathogenic E. coli, Salmonela enterica, Yersinia enterocolitica and Proteus mirabilis, was determined by the DNA sequencing of the conserved 5’ and 3’regions fliC gene (flagellin encoded gene). Among 30 commensal avian E. coli strains and 49 pathogenic avian E. coli strains (APEC), 24 commensal and 39 APEC strains harbored fliC gene with fragments size varying from 670bp to 1,900bp. The comparative analysis of these regions allowed the construction of a dendrogram of similarity possessing two main clusters: one compounded mainly by APEC strains and by H-antigens from human E. coli, and another one compounded by commensal avian E. coli strains, S. enterica, and by other H-antigens from human E. coli. Overall, this work demonstrated that fliC conserved regions may be associated with pathogenic clones of APEC strains, and also shows a great similarity among APEC and H-antigens of E. coli strains isolated from humans. These data, can add evidence that APEC strains can exhibit a zoonotic risk.

Abstract in Portuguese:

ABSTRACT.- Campos T.A., Nakazato G., Stehling E.G., Brocchi M. & Silveira W.D. 2008. Clonal study of avian Escherichia coli strains by fliC conserved-DNA-sequence regions analysis. Pesquisa Veterinária Brasileira 28(10):508-514. Departamento de Microbiologia e Imunologia, Instituto de Biologia, Cx. Postal 6109, Universidade Estadual de Campinas, Cidade Universitária Zeferino Vaz s/n, Barão Geraldo, Campinas, SP 3081-862, Brazil. *Corresponding author: wds@unicamp.br The clonal relationship among avian Escherichia coli strains and their genetic proximity with human pathogenic E. coli, Salmonela enterica, Yersinia enterocolitica and Proteus mirabilis, was determined by the DNA sequencing of the conserved 5’ and 3’regions fliC gene (flagellin encoded gene). Among 30 commensal avian E. coli strains and 49 pathogenic avian E. coli strains (APEC), 24 commensal and 39 APEC strains harbored fliC gene with fragments size varying from 670bp to 1,900bp. The comparative analysis of these regions allowed the construction of a dendrogram of similarity possessing two main clusters: one compounded mainly by APEC strains and by H-antigens from human E. coli, and another one compounded by commensal avian E. coli strains, S. enterica, and by other H-antigens from human E. coli. Overall, this work demonstrated that fliC conserved regions may be associated with pathogenic clones of APEC strains, and also shows a great similarity among APEC and H-antigens of E. coli strains isolated from humans. These data, can add evidence that APEC strains can exhibit a zoonotic risk.


#102 - Occurrence of virulence-related sequences and phylogenetic analysis of commensal and pathogenic avian Escherichia coli strains (APEC), p.533-540

Abstract in English:

ABSTRACT.- Campos T.A., Lago J.C., Nakazato G., Stehling E.G., Brocchi M., Castro A.F.P. & Silveira W.D. 2008. Occurrence of virulence-related sequences and phylogenetic analysis of commensal and pathogenic avian Escherichia coli strains (APEC). Pesquisa Veterinária Brasileira 28(10):533-540. Departamento de Microbiologia e Immunologia, Instituto de Biologia, Unicamp, Cidade Universitrária Zeferino Vaz s/n, Campinas, SP 13081-862, Brazil. E-mail: wds@unicamp.br The presence of iron uptake (irp-2, fyuA, sitA, fepC, iucA), adhesion (iha, lpfAO157/O141, lpfAO157/O154, efa, toxB) and invasion (inv, ial-related DNA sequences and assignment to the four main Escherichia coli phylogenetic groups (A, B1, B2 e D) were determined in 30 commensal E. coli strains isolated from healthy chickens and in 49 APEC strains isolated from chickens presenting clinical signs of septicemia (n=24) swollen head syndrome (n=14) and omphalitis (n=11) by PCR. None of the strains presented DNA sequences related to the inv, ial, efa, and toxB genes. DNA sequences related to lpfAO157/O154, iucA, fepC, and irp-2 genes were significantly found among pathogenic strains, where iucA gene was associated with septicemia and swollen head syndrome and fepC and irp-2 genes were associated with swollen head syndrome strains. Phylogenetic typing showed that commensal and omphalitis strains belonged mainly to phylogenetic Group A and swollen head syndrome to phylogenetic Group D. Septicemic strains were assigned in phylogenetic Groups A and D. These data could suggest that clonal lineage of septicemic APEC strains have a multiple ancestor origin; one from a pathogenic bacteria ancestor and other from a non-pathogenic ancestor that evolved by the acquisition of virulence related sequences through horizontal gene transfer. Swollen head syndrome may constitute a pathogenic clonal group. By the other side, omphalitis strains probably constitute a non-pathogenic clonal group, and could cause omphalitis as an opportunistic infection. The sharing of virulence related sequences by human pathogenic E. coli and APEC strains could indicate that APEC strains could be a source of virulence genes to human strains and could represent a zoonotic risk.

Abstract in Portuguese:

ABSTRACT.- Campos T.A., Lago J.C., Nakazato G., Stehling E.G., Brocchi M., Castro A.F.P. & Silveira W.D. 2008. Occurrence of virulence-related sequences and phylogenetic analysis of commensal and pathogenic avian Escherichia coli strains (APEC). Pesquisa Veterinária Brasileira 28(10):533-540. Departamento de Microbiologia e Immunologia, Instituto de Biologia, Unicamp, Cidade Universitrária Zeferino Vaz s/n, Campinas, SP 13081-862, Brazil. E-mail: wds@unicamp.br The presence of iron uptake (irp-2, fyuA, sitA, fepC, iucA), adhesion (iha, lpfAO157/O141, lpfAO157/O154, efa, toxB) and invasion (inv, ial-related DNA sequences and assignment to the four main Escherichia coli phylogenetic groups (A, B1, B2 e D) were determined in 30 commensal E. coli strains isolated from healthy chickens and in 49 APEC strains isolated from chickens presenting clinical signs of septicemia (n=24) swollen head syndrome (n=14) and omphalitis (n=11) by PCR. None of the strains presented DNA sequences related to the inv, ial, efa, and toxB genes. DNA sequences related to lpfAO157/O154, iucA, fepC, and irp-2 genes were significantly found among pathogenic strains, where iucA gene was associated with septicemia and swollen head syndrome and fepC and irp-2 genes were associated with swollen head syndrome strains. Phylogenetic typing showed that commensal and omphalitis strains belonged mainly to phylogenetic Group A and swollen head syndrome to phylogenetic Group D. Septicemic strains were assigned in phylogenetic Groups A and D. These data could suggest that clonal lineage of septicemic APEC strains have a multiple ancestor origin; one from a pathogenic bacteria ancestor and other from a non-pathogenic ancestor that evolved by the acquisition of virulence related sequences through horizontal gene transfer. Swollen head syndrome may constitute a pathogenic clonal group. By the other side, omphalitis strains probably constitute a non-pathogenic clonal group, and could cause omphalitis as an opportunistic infection. The sharing of virulence related sequences by human pathogenic E. coli and APEC strains could indicate that APEC strains could be a source of virulence genes to human strains and could represent a zoonotic risk.


#103 - Morphological analysis of the corneal endothelium in eyes of dogs using specular microscopy, p.427-430

Abstract in English:

ABSTRACT.- Pigatto J.A.T., Cerva C., Freire C.D., Abib F.C., Bellini L.P., Barros P.S.M. & Laus J.L. 2008. Morphological analysis of the corneal endothelium in eyes of dogs using specular microscopy. Pesquisa Veterinária Brasileira 28(9):427-430. Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: pigatto@ufrgs.br Both healthy eyes of 10 six-year-old male and female mongrel dogs were studied. With a contact specular microscope the corneal endothelium was examined. Endothelial cells were analyzed in the central and peripheral cornea. Morphological analysis with regard to polymegathism and pleomorphism was performed. Three images of each region with at least 100 cells were obtained. The analysis showed that polygonal cells formed a mosaic-like pattern uniform in size and shape. The predominant number of cells was hexagonal. The polymegathism index was 0.22. The study demonstrates that the morphology of the normal corneal endothelial cells of dogs is similar to that found in the human cornea.

Abstract in Portuguese:

ABSTRACT.- Pigatto J.A.T., Cerva C., Freire C.D., Abib F.C., Bellini L.P., Barros P.S.M. & Laus J.L. 2008. Morphological analysis of the corneal endothelium in eyes of dogs using specular microscopy. Pesquisa Veterinária Brasileira 28(9):427-430. Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: pigatto@ufrgs.br Both healthy eyes of 10 six-year-old male and female mongrel dogs were studied. With a contact specular microscope the corneal endothelium was examined. Endothelial cells were analyzed in the central and peripheral cornea. Morphological analysis with regard to polymegathism and pleomorphism was performed. Three images of each region with at least 100 cells were obtained. The analysis showed that polygonal cells formed a mosaic-like pattern uniform in size and shape. The predominant number of cells was hexagonal. The polymegathism index was 0.22. The study demonstrates that the morphology of the normal corneal endothelial cells of dogs is similar to that found in the human cornea.


#104 - Comparison of two commercial kits and two extraction methods for fecal glucocorticoid analysis in ocelots (Leopardus pardalis) submitted to ACTH challenge, p.329-334

Abstract in English:

ABSTRACT.- Dias E.A., Nichi M. & Guimarães M.A.B.V. 2008. Comparison of two commercial kits and two extraction methods for fecal glucocorticoid analysis in ocelots (Leopardus pardalis) submitted to ACTH challenge. Pesquisa Veterinária Brasileira 28(7):329-334. Departamento de Reprodução Animal, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, Cidade Universitária, São Paulo, SP 05508-270, Brazil. E-mail: eantdias@usp.br The ocelot (Leopardus pardalis) is included in list of wild felid species protected by CITES and is part of conservation strategies that necessarily involve the use of assisted reproduction techniques, which requires practical and minimally invasive techniques of high reproducibility that permit the study of animal reproductive physiology. The objective of this study was to compare and validate two commercial assays: ImmuChem Double Antibody Corticosterone 125I RIA from ICN Biomedicals, Costa Mesa, CA, USA; and Coat-a-Count Cortisol 125I RIA from DPC, Los Angeles, CA, USA, for assessment of fecal glucocorticoid metabolites in ocelots submitted to ACTH (adrenocorticotropic hormone) challenge. Fecal samples were collected from five ocelots kept at the Brazilian Center of Neotropical Felines, Associação Mata Ciliar, São Paulo, Brazil, and one of the animals was chosen as a negative control. The experiment was conducted over a period of 9 days. On day 0, a total dose of 100 IU ACTH was administered intramuscularly. Immediately after collection the samples were stored at -20C in labeled plastic bags. The hormone metabolites were subsequently extracted and assayed using the two commercial kits. Previously it was performed a trial with the DPC kit to check the best extraction method for hormones metabolites. Data were analyzed with the SAS program for Windows V8 and reported as means ± SEM. The Schwarzenberger extraction method was slightly better when compared with the Wasser extraction method (103,334.56 ± 19,010.37ng/g of wet feces and 59,223.61 ± 12,725.36ng/g of wet feces respectively; P=0,0657). The ICN kit detected an increase in glucocorticoid metabolite concentrations in a more reliable manner. Metabolite concentrations (ng/g wet feces) on day 0 and day 1 were 66,956.28 ± 36,786.93 and 92,991.19 ± 28,555.63 for the DPC kit, and 205,483.32 ± 83,811.32 and 814,578.75 ± 292,150.47 for the ICN kit, respectively. The limit of detection for the ICN kit was 7.7 ng/mL for 100% B/Bo (25ng/mL for 88%B/Bo) and for the DPC kit it was 0.2ug/dL for 90.95% B/Bo (1ug/dL for 81.27% B/Bo). In conclusion it was confirmed that the Schwarzenberger extraction method and the ICN kit are superior for extracting and measuring fecal glucocorticoid metabolites in ocelot fecal samples.

Abstract in Portuguese:

ABSTRACT.- Dias E.A., Nichi M. & Guimarães M.A.B.V. 2008. Comparison of two commercial kits and two extraction methods for fecal glucocorticoid analysis in ocelots (Leopardus pardalis) submitted to ACTH challenge. Pesquisa Veterinária Brasileira 28(7):329-334. Departamento de Reprodução Animal, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, Cidade Universitária, São Paulo, SP 05508-270, Brazil. E-mail: eantdias@usp.br The ocelot (Leopardus pardalis) is included in list of wild felid species protected by CITES and is part of conservation strategies that necessarily involve the use of assisted reproduction techniques, which requires practical and minimally invasive techniques of high reproducibility that permit the study of animal reproductive physiology. The objective of this study was to compare and validate two commercial assays: ImmuChem Double Antibody Corticosterone 125I RIA from ICN Biomedicals, Costa Mesa, CA, USA; and Coat-a-Count Cortisol 125I RIA from DPC, Los Angeles, CA, USA, for assessment of fecal glucocorticoid metabolites in ocelots submitted to ACTH (adrenocorticotropic hormone) challenge. Fecal samples were collected from five ocelots kept at the Brazilian Center of Neotropical Felines, Associação Mata Ciliar, São Paulo, Brazil, and one of the animals was chosen as a negative control. The experiment was conducted over a period of 9 days. On day 0, a total dose of 100 IU ACTH was administered intramuscularly. Immediately after collection the samples were stored at -20C in labeled plastic bags. The hormone metabolites were subsequently extracted and assayed using the two commercial kits. Previously it was performed a trial with the DPC kit to check the best extraction method for hormones metabolites. Data were analyzed with the SAS program for Windows V8 and reported as means ± SEM. The Schwarzenberger extraction method was slightly better when compared with the Wasser extraction method (103,334.56 ± 19,010.37ng/g of wet feces and 59,223.61 ± 12,725.36ng/g of wet feces respectively; P=0,0657). The ICN kit detected an increase in glucocorticoid metabolite concentrations in a more reliable manner. Metabolite concentrations (ng/g wet feces) on day 0 and day 1 were 66,956.28 ± 36,786.93 and 92,991.19 ± 28,555.63 for the DPC kit, and 205,483.32 ± 83,811.32 and 814,578.75 ± 292,150.47 for the ICN kit, respectively. The limit of detection for the ICN kit was 7.7 ng/mL for 100% B/Bo (25ng/mL for 88%B/Bo) and for the DPC kit it was 0.2ug/dL for 90.95% B/Bo (1ug/dL for 81.27% B/Bo). In conclusion it was confirmed that the Schwarzenberger extraction method and the ICN kit are superior for extracting and measuring fecal glucocorticoid metabolites in ocelot fecal samples.


#105 - Genetic grouping of avian infectious bronchitis virus isolated in Brazil, based on RT-PCR/RFLP analysis of the S1 gene, p.190-194

Abstract in English:

ABSTRACT.- Montassier M.F.S., Brentano L., Montassier H.J. & Richtzenhain L.J. 2008. Genetic grouping of avian infectious bronchitis virus isolated in Brazil, based on RT-PCR/RFLP analysis of the S1 gene. Pesquisa Veterinária Brasileira 28(3):190-194. Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando de Paiva 87, São Paulo, SP 05508-000, Brazil. E-mail: leonardo@usp.br Twelve Brazilian isolates and one reference vaccine strain of avian infectious bronchitis virus (IBV) were propagated in embryonating chicken eggs. The entire S1 glycoprotein gene of these viruses was analysed by reverse-transcriptase-polymerase chain reaction and restriction fragment length polymorphism (RT-PCR-RFLP), using the restriction enzymes HaeIII, XcmI and BstyI. The RFLP patterns led to the classification of these isolates into five distinct genotypes: A, B, C, D and Massachusetts. Five of twelve isolates were grouped in Massachusetts genotype and the remaining seven viruses were classified into four distinct genotypes: A (2), B (2), C (2) or D (1). Such genotyping classification agreed with previous immunological analysis for most of these viruses, highlighting the occurrence of a relevant variability among the IBV strains that are circulating in Brazilian commercial poultry flocks.

Abstract in Portuguese:

ABSTRACT.- Montassier M.F.S., Brentano L., Montassier H.J. & Richtzenhain L.J. 2008. Genetic grouping of avian infectious bronchitis virus isolated in Brazil, based on RT-PCR/RFLP analysis of the S1 gene. Pesquisa Veterinária Brasileira 28(3):190-194. Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando de Paiva 87, São Paulo, SP 05508-000, Brazil. E-mail: leonardo@usp.br Twelve Brazilian isolates and one reference vaccine strain of avian infectious bronchitis virus (IBV) were propagated in embryonating chicken eggs. The entire S1 glycoprotein gene of these viruses was analysed by reverse-transcriptase-polymerase chain reaction and restriction fragment length polymorphism (RT-PCR-RFLP), using the restriction enzymes HaeIII, XcmI and BstyI. The RFLP patterns led to the classification of these isolates into five distinct genotypes: A, B, C, D and Massachusetts. Five of twelve isolates were grouped in Massachusetts genotype and the remaining seven viruses were classified into four distinct genotypes: A (2), B (2), C (2) or D (1). Such genotyping classification agreed with previous immunological analysis for most of these viruses, highlighting the occurrence of a relevant variability among the IBV strains that are circulating in Brazilian commercial poultry flocks.


#106 - Detection and phylogenetic analysis of porcine enteric calicivirus, genetically related to the Cowden strain of sapovirus genogroup III, in Brazilian swine herds, p.82-86

Abstract in English:

Abstract.- Barry A.F, Alfieri A.F. & Alfieri A.A. 2008. Detection and phylogenetic analysis of porcine enteric calicivirus, genetically related to the Cowden strain of sapovirus genogroup III, in Brazilian swine herds. Pesquisa Veterinária Brasileira 28(1):82-86. Laboratório de Virologia Animal, Departamento de Medicina Veterinária Preventiva, Centro de Ciências Agrárias, Universidade Estadual de Londrina, Campus Universitário, Londrina, PR 86051-990, Brazil. E-mail: alinebarry@uol.com.br Sapovirus of the Caliciviridae family is an important agent of acute gastroenteritis in children and piglets. The Sapovirus genus is divided into seven genogroups (G), and strains from the GIII, GVI and GVII are associated with infections in swine. Despite the high prevalence in some countries, there are no studies related to the presence of porcine enteric sapovirus infections in piglets in Brazil. In the present study, 18 fecal specimens from piglets up to 28 days were examined to determine the presence of sapovirus genome by RT-PCR assay, using primers designed to amplify a 331 bp segment of the RNA polymerase gene. In 44.4% (8/18) of fecal samples, an amplified DNA fragment was obtained. One of these fragments was sequenced and submitted to molecular and phylogenetic analysis. This analysis revealed high similarity, with nucleotides (87%) and amino acids (97.8%), to the Cowden strain, the GIII prototype of porcine enteric calicivirus. This is the first description of sapovirus in Brazilian swine herds.

Abstract in Portuguese:

Abstract.- Barry A.F, Alfieri A.F. & Alfieri A.A. 2008. Detection and phylogenetic analysis of porcine enteric calicivirus, genetically related to the Cowden strain of sapovirus genogroup III, in Brazilian swine herds. Pesquisa Veterinária Brasileira 28(1):82-86. Laboratório de Virologia Animal, Departamento de Medicina Veterinária Preventiva, Centro de Ciências Agrárias, Universidade Estadual de Londrina, Campus Universitário, Londrina, PR 86051-990, Brazil. E-mail: alinebarry@uol.com.br Sapovirus of the Caliciviridae family is an important agent of acute gastroenteritis in children and piglets. The Sapovirus genus is divided into seven genogroups (G), and strains from the GIII, GVI and GVII are associated with infections in swine. Despite the high prevalence in some countries, there are no studies related to the presence of porcine enteric sapovirus infections in piglets in Brazil. In the present study, 18 fecal specimens from piglets up to 28 days were examined to determine the presence of sapovirus genome by RT-PCR assay, using primers designed to amplify a 331 bp segment of the RNA polymerase gene. In 44.4% (8/18) of fecal samples, an amplified DNA fragment was obtained. One of these fragments was sequenced and submitted to molecular and phylogenetic analysis. This analysis revealed high similarity, with nucleotides (87%) and amino acids (97.8%), to the Cowden strain, the GIII prototype of porcine enteric calicivirus. This is the first description of sapovirus in Brazilian swine herds.


#107 - IgA production, coliforms analysis and intestinal mucosa morphology of piglets that received probiotics with viable or inactivated cells, p.241-245

Abstract in English:

ABSTRACT.- Rodrigues M.A.M., Oliveira D.A., Taketomi E.A. & Hernandez-Blazquez F.J. 2007. IgA production, coliforms analysis and intestinal mucosa morphology of piglets that received probiotics with viable or inactivated cells. Pesquisa Veterinária Brasileira 27(6):241-245. Departamento de Cirurgia, Setor de Anatomia, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, São Paulo, SP 05508-900, Brazil. Email: fjhblazq@usp.br Two types of probiotics were used in piglets. One product is a mixed culture of viable Lactobacillus acidophilus, Enterococcus faecium e Bifidobacterium bifidum. The second product is composed of inactivated Lactobacillus acidophilus cells. The piglets received two weekly oral doses for 30 days while a control group did not receive probiotics. All piglets were euthanized at the 30th day of life and the mesenteric lymph nodes, the small intestine, and blood samples were collected. The tissue samples were studied by light microscopy and the blood serum was analyzed by ELISA method. The treatment with the probiotic with viable cells produced higher serum levels of IgA (P<0.05) and more IgA expressing cells were found in the mesenteric lymph nodes than observed in the inactivated cells treatment or control groups (P<0.05). Also, intestinal villi were longer, crypts were deeper (P<0.05) and fecal coliform count was lower than found in the inactivated product (P<0.05). These results suggest that viable probiotics are more efficient than inactivated probiotics to induce immunostimulation and intestinal modifications in piglets, thus improving their health and development.

Abstract in Portuguese:

ABSTRACT.- Rodrigues M.A.M., Oliveira D.A., Taketomi E.A. & Hernandez-Blazquez F.J. 2007. IgA production, coliforms analysis and intestinal mucosa morphology of piglets that received probiotics with viable or inactivated cells. Pesquisa Veterinária Brasileira 27(6):241-245. Departamento de Cirurgia, Setor de Anatomia, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, São Paulo, SP 05508-900, Brazil. Email: fjhblazq@usp.br Two types of probiotics were used in piglets. One product is a mixed culture of viable Lactobacillus acidophilus, Enterococcus faecium e Bifidobacterium bifidum. The second product is composed of inactivated Lactobacillus acidophilus cells. The piglets received two weekly oral doses for 30 days while a control group did not receive probiotics. All piglets were euthanized at the 30th day of life and the mesenteric lymph nodes, the small intestine, and blood samples were collected. The tissue samples were studied by light microscopy and the blood serum was analyzed by ELISA method. The treatment with the probiotic with viable cells produced higher serum levels of IgA (P<0.05) and more IgA expressing cells were found in the mesenteric lymph nodes than observed in the inactivated cells treatment or control groups (P<0.05). Also, intestinal villi were longer, crypts were deeper (P<0.05) and fecal coliform count was lower than found in the inactivated product (P<0.05). These results suggest that viable probiotics are more efficient than inactivated probiotics to induce immunostimulation and intestinal modifications in piglets, thus improving their health and development.


#108 - Genotype characterization of the horn fly Haematobia irritans from different Brazilian geographic regions based on randomly amplified polymorphic DNA (RAPD) analysis, p.1-5

Abstract in English:

ABSTRACT.- Brito L.G., Regitano L.C.A., Huacca M.E.F., Carrilho E. & Moya Borja G.E. 2007. Genotype characterization of the horn fly Haematobia irritans from different Brazilian geographic regions based on randomly amplified polymorphic DNA (RAPD) analysis. Pesquisa Veterinária Brasileira 27(1):1-5. Laboratório de Sanidade Animal, Embrapa Rondônia, BR 364 Km 5,5, Porto Velho, RO 78900-970, Brazil. E-mail: luciana@cpafro.embrapa.br Blood-sucking diptera are important parasites in bovine production systems, especially regarding confinement conditions. Haematobia irritans, the horn fly, is one of the most troublesome species within bovine production systems, due to the intense stress imposed to the animals. An important aspect while studying the variability within a species is the study of the geographic structure of its populations and, attempting to find out the genetic flow of Brazilian populations of horn fly, the RAPD technique, which is suited for this purpose, has been used. The use of molecular markers generated from RAPD made it possible to identify the geographic origin of samples from different Brazilian geographic regions, as well as to estimate the genotypic flow among the different Brazilian populations of the horn fly.

Abstract in Portuguese:

ABSTRACT.- Brito L.G., Regitano L.C.A., Huacca M.E.F., Carrilho E. & Moya Borja G.E. 2007. Genotype characterization of the horn fly Haematobia irritans from different Brazilian geographic regions based on randomly amplified polymorphic DNA (RAPD) analysis. Pesquisa Veterinária Brasileira 27(1):1-5. Laboratório de Sanidade Animal, Embrapa Rondônia, BR 364 Km 5,5, Porto Velho, RO 78900-970, Brazil. E-mail: luciana@cpafro.embrapa.br Blood-sucking diptera are important parasites in bovine production systems, especially regarding confinement conditions. Haematobia irritans, the horn fly, is one of the most troublesome species within bovine production systems, due to the intense stress imposed to the animals. An important aspect while studying the variability within a species is the study of the geographic structure of its populations and, attempting to find out the genetic flow of Brazilian populations of horn fly, the RAPD technique, which is suited for this purpose, has been used. The use of molecular markers generated from RAPD made it possible to identify the geographic origin of samples from different Brazilian geographic regions, as well as to estimate the genotypic flow among the different Brazilian populations of the horn fly.


#109 - Genetic characterization of Brazilian bovine viral diarrhea virus isolates by partial nucleotide sequencing of the 5’-UTR region

Abstract in English:

ABSTRACT.- Cortez A., Heinemann M.B., Castro A.M.M.G., Soares M.S, Pinto A.M.V., Alfieri A.A., Flores E.F., Leite R.C. & Richtzenhain L.J. 2006. Genetic characterization of Brazilian bovine viral diarrhea virus isolates by partial nucleotide sequencing of the 5’-UTR region. Pesquisa Veterinária Brasileira 26(4):211-216. Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, Cidade Universitária, São Paulo, SP 05508-000, Brazil. E-mail: leonardo@usp.br Nineteen isolates of bovine viral diarrhea virus (BVDV) from Brazil were genetically characterized through partial nucleotide sequencing and analysis of the 5’UTR region. The isolates were grouped as BVDV-1 (11/19), BVDV-2 (6/19) or “atypical” pestivirus (2/19). Among the BVDV-1, eight isolates were classified as subgenotype BVDV-1a, whereas most (4 out of 6) BVDV-2 belonged to subgenotype 2b. Two isolates from aborted fetuses were not classified into any genetic group, being considered atypical BVDVs. Genetic diversity among Brazilian BVDV isolates may be responsible for vaccination and diag-nostic failure and therefore may influence the control strategies for BVDV infection in the country.

Abstract in Portuguese:

ABSTRACT.- Cortez A., Heinemann M.B., Castro A.M.M.G., Soares M.S, Pinto A.M.V., Alfieri A.A., Flores E.F., Leite R.C. & Richtzenhain L.J. 2006. Genetic characterization of Brazilian bovine viral diarrhea virus isolates by partial nucleotide sequencing of the 5’-UTR region. Pesquisa Veterinária Brasileira 26(4):211-216. Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, Cidade Universitária, São Paulo, SP 05508-000, Brazil. E-mail: leonardo@usp.br Nineteen isolates of bovine viral diarrhea virus (BVDV) from Brazil were genetically characterized through partial nucleotide sequencing and analysis of the 5’UTR region. The isolates were grouped as BVDV-1 (11/19), BVDV-2 (6/19) or “atypical” pestivirus (2/19). Among the BVDV-1, eight isolates were classified as subgenotype BVDV-1a, whereas most (4 out of 6) BVDV-2 belonged to subgenotype 2b. Two isolates from aborted fetuses were not classified into any genetic group, being considered atypical BVDVs. Genetic diversity among Brazilian BVDV isolates may be responsible for vaccination and diag-nostic failure and therefore may influence the control strategies for BVDV infection in the country.


#110 - Genetic and antigenic analysis of Babesia bigemina isolates from five geographical regions of Brazil, 22(4):153-160

Abstract in English:

ABSTRACT.- Madruga C.R., Leal C.R.B., Ferreira A.M.T., Araújo F.R., Bonato A.L.V., Kessler R.H., Schenk M.A.M. & Soares C.O. 2002. Genetic and antigenic analysis of Babesia bigemina isolates from five geographical regions of Brazil. Pesquisa Veterinária Brasileira 22(4):153- 160. [Análise genética e antigênica de isolados de Babesia bigemina das cinco regiões fisiográficas do Brasil.] Embrapa Gado de Corte, Rodovia BR 262 Km 4, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. A molecular epidemiological study was performed with Babesia bigemina isolates from five geographical regions of Brazil. The genetic analysis was done with random amplification of polymorphic DNA (RAPD), repetitive extragenic palindromic elements-polymerase chain reaction (REP-PCR) and enterobacterial repetitive intergenic consensus sequences-polymerase chain reaction (ERIC-PCR) that showed genetic polymorphism between these isolates and generated fingerprinting. In RAPD, IL0872 and IL0876 primers were able to detect at least one fingerprinting for each B. bigemina isolate. The amplification of B. bigemina DNA fragments by REP-PCR and ERIC-PCR gave evidence for the presence in this haemoprotozoan of the sequences described previously in microorganisms of the bacterial kingdom. For the first time it was demonstrated that both techniques can be used for genetic analysis of a protozoan parasite, although the ERIC-PCR was more discriminatory than REP-PCR. Toe dendogram with similarity coeficiente among isolates showed two clusters and one subcluster. The Northeastern and Mid-Westem isolates showed the greatest genetic diversity, while the Southeastem and Southem isolates were the closest. Toe antigenic analysis was done through indirect fluorescent antibodytechnique and Westem blotting using a panei of monoclonal antibodies directed against epitopes on the merozoite membrane surface, rhoptries and membrane of infected erythrocytes. As expected, the merozoite variable surface antigens, major surface antigen (MSA)-1 and MSA-2 showed antigenic diversity. However, B cell epitopes on rhoptries and infected erythrocytes were conserved among all isolates studied. In this study it was possible to identify variable and conserved antigens, which had already been described as potential immunogens. Considering that an attenuated Babesia clone used as immunogen selected populations capable of evading the immunity induced by this vaccine, it is necessary to evaluate more deeply the cross-protection conferred by genetically more distant Brazilian B. bigemina isolates and make an evaluation of the polymorphism degree of variable antigens such as MSA-1 and MSA-2.

Abstract in Portuguese:

RESUMO.- Madruga C.R., Leal C.R.B., Ferreira A.M.T., Araújo F.R., Bonato A.L.V., Kessler R.H., Schenk M.A.M. & Soares C.O. 2002. Genetic and antigenic analysis of Babesia bigemina isolates from five geographical regions of Brazil. Pesquisa Veterinária Brasileira 22(4):153- 160. [Análise genética e antigênica de isolados de Babesia bigemina das cinco regiões fisiográficas do Brasil.] Embrapa Gado de Corte, Rodovia BR 262 Km 4, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. Um estudo de epidemiologia molecular foi executado com isolados de Babesia bigemina das cinco regiões fisiográficas do Brasil. A análise genética foi feita com amplificação aleatória de DNA polimórfico (RAPD), reação da polimerase em cadeia com seqüências de elementos extragênicos repetitivos palindrômicos (REP-PCR) e reação da polimerase em cadeia com seqüências repetitivas enterobacterianas intergênicas de consenso (ERIC-PCR) que apresentaram polimorfismo genético entre os isolados e geraram marcadores. No RAPD com os oligonucleotídeos iniciadores IL0872 e IL0876, foi possível detectar pelo menos um marcador por isolado de B. bigemina. A amplificação de fragmentos de DNA de B. bigemina por REPPCR e ERIC-PCR demonstrou a presença dessas seqüências, descritas anteriormente somente em microrganismos bacterianos, nesse hemoprotozoârio, e, pela primeira vez, foi verificado que podem ser utilizadas para análise genética de um protozoário. O ERIC-PCR foi mais discriminatório que o REP-PCR. O dendograma formado com o coeficiente de similaridade entre os isolados evidenciou dois agrupamentos e um subgrupo. Os isolados do Nordeste e Centro-Oeste demonstraram maior diversidade genética, enquanto que os isolados do Sudeste e Sul foram os mais próximos. A análise antigênica foi executada por meio de imunofluorescência indireta e Western blotting usando um painel de anticorpos monoclonais direcionados a epitopos B na membrana dos merozoítos, roptries e membrana de eritrócitos infectados. Os antígenos variáveis da superfície dos merozoítos, antígeno principal da superfície do merozoíto (APSM)-1 e APSM-2 apresentaram diversidade antigênica. Entretanto, os epítopos de células B nas roptries e nos eritrócitos infectados foram conservados em todos os isolados. Nesse estudo foi possível identificar antígenos variáveis e conservados que anteriormente haviam sido descritos como potenciais imunógenos. Considerando que um clone atenuado de Babesia utilizado para imunização selecionou populações capazes de evadir a resposta imune à vacina, torna-se necessário avaliar mais detalhadamente a imunidade cruzada existente entre os isolados brasileiros mais distantes geneticamente e realizar uma avaliação do grau de polimorfismo dos antígenos variáveis APSM-1 e APSM-2.


Colégio Brasileiro de Patologia Animal SciELO Brasil CAPES CNPQ UFRRJ CFMV