PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Moraes J.M., Brito L.A.B., Moura V.M.B.D., Ribeiro C.S., Guimarães V.Y., Andrade D.F., Lobo J.R. & Fioravanti M.C.S. 2009. [Immunophenotyping and quantitative evaluation of circulating lymphocytes of cattle of the Curraleiro breed.] Imunofenotipagem e avaliação quantitativa de linfócitos circulantes de bovinos da raça Curraleiro. Pesquisa Veterinária Brasileira 29(4):339-344. Escola de Veterinária, Universidade Federal de Goiás, Campus Samambaia, Cx. Postal 131, Goiânia, GO 74001-970, Brazil. E-mail: mmjulia_vet@yahoo.com.br The aim of this study was to establish an immunological profile by marking and quantification of T and B lymphocytes in Curraleiro breed by immunocytochemistry. A total of 116 cattle was used, males and females of different ages, from two farms situated in the Goiás State, Brazil. The animals were allotted into groups according to age, sex and origin. Blood samples were collected and processed in accordance with immunocytochemistry standard technique using lymphoid markers species-specific, as anti-CD3 (MM1A - BoCD3) and anti-LB (LCTB16A-clone B-B14), for T and B lymphocyte counting, respectively. The data were submitted to descriptive statistics, and to Kruskall Wallis and Mann-Whitney tests. The results showed decreased levels of leukocytes, lymphocytes, T lymphocytes and B lymphocytes along the age advance. Absolute values of leukocytes, lymphocytes and T lymphocytes were higher in males than females. None of the evaluated parameters were affected by differences of the management carried out at the two farms.

• Immunocytochemistry CD3 T lymphocytes B lymphocytes immunological profile

Abstract in Portuguese:

ABSTRACT.- Moraes J.M., Brito L.A.B., Moura V.M.B.D., Ribeiro C.S., Guimarães V.Y., Andrade D.F., Lobo J.R. & Fioravanti M.C.S. 2009. [Immunophenotyping and quantitative evaluation of circulating lymphocytes of cattle of the Curraleiro breed.] Imunofenotipagem e avaliação quantitativa de linfócitos circulantes de bovinos da raça Curraleiro. Pesquisa Veterinária Brasileira 29(4):339-344. Escola de Veterinária, Universidade Federal de Goiás, Campus Samambaia, Cx. Postal 131, Goiânia, GO 74001-970, Brazil. E-mail: mmjulia_vet@yahoo.com.br The aim of this study was to establish an immunological profile by marking and quantification of T and B lymphocytes in Curraleiro breed by immunocytochemistry. A total of 116 cattle was used, males and females of different ages, from two farms situated in the Goiás State, Brazil. The animals were allotted into groups according to age, sex and origin. Blood samples were collected and processed in accordance with immunocytochemistry standard technique using lymphoid markers species-specific, as anti-CD3 (MM1A - BoCD3) and anti-LB (LCTB16A-clone B-B14), for T and B lymphocyte counting, respectively. The data were submitted to descriptive statistics, and to Kruskall Wallis and Mann-Whitney tests. The results showed decreased levels of leukocytes, lymphocytes, T lymphocytes and B lymphocytes along the age advance. Absolute values of leukocytes, lymphocytes and T lymphocytes were higher in males than females. None of the evaluated parameters were affected by differences of the management carried out at the two farms.

Abstract in English:

ABSTRACT.- Santos-Rodrigues A., Dagli M.L.Z., Avanzo J.L., Moraes H.P., Mackowiak I.I. & Hernandez-Blazquez F.J. 2009. Expression and distribution of connexin 32 in rat liver with experimentally induced fibrosis. Pesquisa Veterinária Brasileira 29(4):353-357. 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 05508-270, Brazil. E-mail: alexsantos@usp.br The connexin 32 (Cx32) is a protein that forms the channels that promote the gap junction intercellular communication (GJIC) in the liver, allowing the diffusion of small molecules through cytosol from cell-to-cell. Hepatic fibrosis is characterized by a disruption of normal tissue architeture by cellular lesions, and may alter the GJIC. This work aimed to study the expression and distribution of Cx32 in liver fibrosis induced by the oral administration of dimethylnitrosamine in female Wistar rats. The necropsy of the rats was carried out after five weeks of drug administration. They presented a hepatic fibrosis state. Sections from livers with fibrosis and from control livers were submitted to immunohistochemical, Real Time-PCR and Western-Blot analysis to Cx32. In fibrotic livers the Cxs were diffusely scattered in the cytoplasm, contrasting with the control livers, where the Cx32 formed junction plaques at the cell membrane. Also it was found a decrease in the gene expression of Cx32 without reduction in the protein quantity when compared with controls. These results suggest that there the mechanism of intercellular communication between hepatocytes was reduced by the fibrotic process, which may predispose to the occurrence of a neoplastic process, taken in account that connexins are considered tumor suppressing genes.

• Dimethylnitrosamine fibrosis liver Cx32 gap junctions

Abstract in Portuguese:

ABSTRACT.- Santos-Rodrigues A., Dagli M.L.Z., Avanzo J.L., Moraes H.P., Mackowiak I.I. & Hernandez-Blazquez F.J. 2009. Expression and distribution of connexin 32 in rat liver with experimentally induced fibrosis. Pesquisa Veterinária Brasileira 29(4):353-357. 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 05508-270, Brazil. E-mail: alexsantos@usp.br The connexin 32 (Cx32) is a protein that forms the channels that promote the gap junction intercellular communication (GJIC) in the liver, allowing the diffusion of small molecules through cytosol from cell-to-cell. Hepatic fibrosis is characterized by a disruption of normal tissue architeture by cellular lesions, and may alter the GJIC. This work aimed to study the expression and distribution of Cx32 in liver fibrosis induced by the oral administration of dimethylnitrosamine in female Wistar rats. The necropsy of the rats was carried out after five weeks of drug administration. They presented a hepatic fibrosis state. Sections from livers with fibrosis and from control livers were submitted to immunohistochemical, Real Time-PCR and Western-Blot analysis to Cx32. In fibrotic livers the Cxs were diffusely scattered in the cytoplasm, contrasting with the control livers, where the Cx32 formed junction plaques at the cell membrane. Also it was found a decrease in the gene expression of Cx32 without reduction in the protein quantity when compared with controls. These results suggest that there the mechanism of intercellular communication between hepatocytes was reduced by the fibrotic process, which may predispose to the occurrence of a neoplastic process, taken in account that connexins are considered tumor suppressing genes.

Abstract in English:

ABSTRACT.- Silva Júnior A., Castro L.A., Chiarelli Neto O., Silva F.M.F., Vidigal P.M.P., Moraes M.P. & Almeida M.R. 2009. Development and evaluation of a recombinant DNA vaccine candidate expressing porcine circovirus 2 structural protein. Pesquisa Veterinária Brasileira 29(1):76-82. Laboratório de Infectologia Molecular Animal, Instituto de Biotecnologia Aplicada à Agropecuária, Universidade Federal de Viçosa, Av. PH Rolfs s/n, Campus Universitário, Viçosa, MG 36570-000, Brazil. E-mail: marcia@ufv.br Porcine circovirus 2 (PCV2) is generally associated with the porcine circovirosis syndrome, which is considered an important disease of swine and has potentially serious economic impact on the swine industry worldwide. This article describes the construction of a recombinant plasmid expressing the PCV2 structural protein and the evaluation of cellular and humoral immune responses produced by this recombinant vaccine in BALB/c mice. The vaccine candidate was obtained and analyzed in vivo, in an effort to determine the ability to induce a specific immune response in mice. DNA was extracted from a Brazilian PCV2 isolate and the gene coding for Cap protein was amplified by PCR and inserted into an expression plasmid. Groups of BALB/c mice were inoculated intra-muscularly and intradermally in a 15-day interval, with 100 µg and 50 µg of the vaccine construct, respectively. Another group was inoculated intramuscularly with 100 µg of empty plasmid, corresponding to the control group. Seroconversion and cellular response in BALB/c mice were compared and used for vaccine evaluation. Seroconversion was analyzed by ELISA. After a series of 3 immunizations the spleen cells of the immunized animals were used to perform lymphocyte proliferation assays. Seroconversion to PCV2 was detected by ELISA in the animals inoculated with the vaccine construct when compared with control groups. Lymphocyte proliferation assays showed a stronger cell proliferation in the inoculated animals compared with the control group. Thus, the vaccine candidate construct demonstrated to be able to induce both humoral and cellular responses in inoculated mice.

• PCV2 DNA vaccine porcine circovirosis

Abstract in Portuguese:

ABSTRACT.- Silva Júnior A., Castro L.A., Chiarelli Neto O., Silva F.M.F., Vidigal P.M.P., Moraes M.P. & Almeida M.R. 2009. Development and evaluation of a recombinant DNA vaccine candidate expressing porcine circovirus 2 structural protein. Pesquisa Veterinária Brasileira 29(1):76-82. Laboratório de Infectologia Molecular Animal, Instituto de Biotecnologia Aplicada à Agropecuária, Universidade Federal de Viçosa, Av. PH Rolfs s/n, Campus Universitário, Viçosa, MG 36570-000, Brazil. E-mail: marcia@ufv.br Porcine circovirus 2 (PCV2) is generally associated with the porcine circovirosis syndrome, which is considered an important disease of swine and has potentially serious economic impact on the swine industry worldwide. This article describes the construction of a recombinant plasmid expressing the PCV2 structural protein and the evaluation of cellular and humoral immune responses produced by this recombinant vaccine in BALB/c mice. The vaccine candidate was obtained and analyzed in vivo, in an effort to determine the ability to induce a specific immune response in mice. DNA was extracted from a Brazilian PCV2 isolate and the gene coding for Cap protein was amplified by PCR and inserted into an expression plasmid. Groups of BALB/c mice were inoculated intra-muscularly and intradermally in a 15-day interval, with 100 µg and 50 µg of the vaccine construct, respectively. Another group was inoculated intramuscularly with 100 µg of empty plasmid, corresponding to the control group. Seroconversion and cellular response in BALB/c mice were compared and used for vaccine evaluation. Seroconversion was analyzed by ELISA. After a series of 3 immunizations the spleen cells of the immunized animals were used to perform lymphocyte proliferation assays. Seroconversion to PCV2 was detected by ELISA in the animals inoculated with the vaccine construct when compared with control groups. Lymphocyte proliferation assays showed a stronger cell proliferation in the inoculated animals compared with the control group. Thus, the vaccine candidate construct demonstrated to be able to induce both humoral and cellular responses in inoculated mice.

Abstract in English:

ABSTRACT.- Juvenal J.C., Erdmann R.H., Moreira N., Moraes W., Cubas P.H., Delgado L.E.S., Carvalho A.L. & Pachaly J.R. 2008. [Chemical restraint of tigrinas, Leopardus tigrinus, for semen collection with allometrically scaled doses of tiletamine, zolazepam, and xylazine.] Contenção farmacológica de gato-do-mato-pequeno, Leopardus tigrinus, para colheita de sêmen, pela associação de tiletamina zolazepam e xilazina. Pesquisa Veterinária Brasileira 28(11):541-546. Curso de Medicina Veterinária, Universidade Pa-ranaense, Cx. Postal 162, Umuarama, PR 87502-970, Brazil. E-mail: pachaly@uol.com.br This paper reports the anesthetic effects of the combination of tiletamine HCl, zolazepam HCl, and xylazine HCl in tigrinas, Leopardus tigrinus Schreber, 1775 (Fam. Felidae), submitted to semen collection by electroejaculation. Three different protocols and the individual anesthetic doses were calculated by interspecific allometric scaling, based on the usual recommendations for a 10.0 kg domestic dog: On Protocol 1 (n=10) the basis for calculation was 5.0mg/kg for tiletamine + zolazepam and 0.5mg/kg for xylazine; on Protocol 2 (n=12) 5.0mg/kg for tiletamine + zolazepam and 0.75mg/kg for xylazine; and on Protocol 3 (n=11) 5.0mg/kg for tiletamine + zolazepam and 1.0mg/kg for xylazine. The tigrinas were anesthetized on three different occasions with a minimum interval of 30 days. During 120 minutes after the drug administration cardiac and respiratory frequencies, rectal temperature, limb myorelaxation and sensitivity to deep pain were monitored. Latency period, anesthetic period, and contamination of the semen with urine were also monitored. From a total of 32 collections, 10 samples (31.2%) and 18 aliquots (0.07%) were contaminated and rejected, but this episodes were not detrimental for semen analysis and processing. A discrete increase in rectal temperature during electroejaculation caused by muscle contraction, followed by temperature decrease, was observed. Cardiac and respiratory frequency varied during the experiment, but remained within physiological standards for the species. The three tested protocols showed to be safe and effective to produce analgesia and anesthesia in L. tigrinus during semen collection by electroejaculation.

• Chemical restraint anesthesia tigrina Leopardus tigrinus allometric scaling semen collection

Abstract in Portuguese:

ABSTRACT.- Juvenal J.C., Erdmann R.H., Moreira N., Moraes W., Cubas P.H., Delgado L.E.S., Carvalho A.L. & Pachaly J.R. 2008. [Chemical restraint of tigrinas, Leopardus tigrinus, for semen collection with allometrically scaled doses of tiletamine, zolazepam, and xylazine.] Contenção farmacológica de gato-do-mato-pequeno, Leopardus tigrinus, para colheita de sêmen, pela associação de tiletamina zolazepam e xilazina. Pesquisa Veterinária Brasileira 28(11):541-546. Curso de Medicina Veterinária, Universidade Pa-ranaense, Cx. Postal 162, Umuarama, PR 87502-970, Brazil. E-mail: pachaly@uol.com.br This paper reports the anesthetic effects of the combination of tiletamine HCl, zolazepam HCl, and xylazine HCl in tigrinas, Leopardus tigrinus Schreber, 1775 (Fam. Felidae), submitted to semen collection by electroejaculation. Three different protocols and the individual anesthetic doses were calculated by interspecific allometric scaling, based on the usual recommendations for a 10.0 kg domestic dog: On Protocol 1 (n=10) the basis for calculation was 5.0mg/kg for tiletamine + zolazepam and 0.5mg/kg for xylazine; on Protocol 2 (n=12) 5.0mg/kg for tiletamine + zolazepam and 0.75mg/kg for xylazine; and on Protocol 3 (n=11) 5.0mg/kg for tiletamine + zolazepam and 1.0mg/kg for xylazine. The tigrinas were anesthetized on three different occasions with a minimum interval of 30 days. During 120 minutes after the drug administration cardiac and respiratory frequencies, rectal temperature, limb myorelaxation and sensitivity to deep pain were monitored. Latency period, anesthetic period, and contamination of the semen with urine were also monitored. From a total of 32 collections, 10 samples (31.2%) and 18 aliquots (0.07%) were contaminated and rejected, but this episodes were not detrimental for semen analysis and processing. A discrete increase in rectal temperature during electroejaculation caused by muscle contraction, followed by temperature decrease, was observed. Cardiac and respiratory frequency varied during the experiment, but remained within physiological standards for the species. The three tested protocols showed to be safe and effective to produce analgesia and anesthesia in L. tigrinus during semen collection by electroejaculation.

Abstract in English:

ABSTRACT.- Rocha A.C.G.P., Rocha S.L.S., Lima-Rosa C.A.V., Souza G.F., Moraes H.L.S., Salle F.O., Moraes L.B. & Salle C.T.P. 2008. Genes associated with pathogenicity of avian Escherichia coli (APEC) isolated from respiratory cases of poultry. Pesquisa Veterinária Brasileira 28(3):183-186. Centro de Diagnóstico e Pesquisa em Patologia Aviária, Departamento de Medicina Animal, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 8824, Porto Alegre, RS 91540-000, Brazil. E-mail: ana.crocha@terra.com.br The virulence mechanisms of avian pathogenic Escherichia coli (APEC) have been continually studied and are believed to be multi-factorial. Certain properties are primarily associated with virulent samples and have been identified in avian isolates. In this study a total of 61 E. coli, isolates from chicken flocks with respiratory symptomatology, were probed by Polimerase Chain Reation (PCR) for the presence of genes responsible for the adhesion capacity, P fimbria (papC) e F11 fimbria (felA), colicin production (cvaC), aerobactin presence (iutA), serum resistance (iss), temperature-sensitive hemagglutinin (tsh), and presence of K1 and K5 capsular antigens (kpsII). The iss gene was detected in 73,8%, tsh in 55,7%, iutA in 45,9%, felA in 39,3%, papC in 24,3%, cvaC in 23% and kpsII in18%.

• Escherichia coli virulence PCR poultry pathogenicity

Abstract in Portuguese:

ABSTRACT.- Rocha A.C.G.P., Rocha S.L.S., Lima-Rosa C.A.V., Souza G.F., Moraes H.L.S., Salle F.O., Moraes L.B. & Salle C.T.P. 2008. Genes associated with pathogenicity of avian Escherichia coli (APEC) isolated from respiratory cases of poultry. Pesquisa Veterinária Brasileira 28(3):183-186. Centro de Diagnóstico e Pesquisa em Patologia Aviária, Departamento de Medicina Animal, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 8824, Porto Alegre, RS 91540-000, Brazil. E-mail: ana.crocha@terra.com.br The virulence mechanisms of avian pathogenic Escherichia coli (APEC) have been continually studied and are believed to be multi-factorial. Certain properties are primarily associated with virulent samples and have been identified in avian isolates. In this study a total of 61 E. coli, isolates from chicken flocks with respiratory symptomatology, were probed by Polimerase Chain Reation (PCR) for the presence of genes responsible for the adhesion capacity, P fimbria (papC) e F11 fimbria (felA), colicin production (cvaC), aerobactin presence (iutA), serum resistance (iss), temperature-sensitive hemagglutinin (tsh), and presence of K1 and K5 capsular antigens (kpsII). The iss gene was detected in 73,8%, tsh in 55,7%, iutA in 45,9%, felA in 39,3%, papC in 24,3%, cvaC in 23% and kpsII in18%.

Abstract in English:

ABSTRACT.- Aguiar D.M, Gennari S.M., Cavalcante G.T., Labruna M.B., Vasconcellos S.A., Rodrigues A.A.R., Moraes Z.M. & Camargo L.M.A. 2006. Seroprevalence of Leptospira spp in cattle from Monte Negro municipality, western Amazon. Pesquisa Veterinária Brasileira 26(2):102-104. Department of Preventive Veterinary Medicine and Animal Health, Faculty of Veterinary Medicine and Animal Production, University of São Paulo, Av. Prof. Orlando Marques de Paiva 87, São Paulo, SP 05508-900, Brazil. E-mail: danmoura@aptaregional.sp.gov.br The prevalence of anti-Leptospira spp antibodies was investigated in 2,109 female cattle from 86 herds of Monte Negro municipality, Rondônia, Brazil. Sera samples were evaluated by Microscopic Agglutination Test against 24 leptospira serovars. Titers =100 for at least one of 24 leptospira serovars were detected in 1,114 cows (52.8%) from 82 (95.3%) herds. The adjusted overall prevalence for Monte Negro municipality was 53.9% (49-58.7%; CI: 95%). The most prevalent serovars were Hardjo (14.5%), Wolffi (12.3%), Shermani (10.8%), Patoc (7.9%), and Hebdomadis (6.1%). Other serovars worldwidely reported like Bratislava, Pomona and Grippotyphosa were detected in low levels.

• Leptospira spp cattle epidemiology Amazon.

Abstract in Portuguese:

ABSTRACT.- Aguiar D.M, Gennari S.M., Cavalcante G.T., Labruna M.B., Vasconcellos S.A., Rodrigues A.A.R., Moraes Z.M. & Camargo L.M.A. 2006. Seroprevalence of Leptospira spp in cattle from Monte Negro municipality, western Amazon. Pesquisa Veterinária Brasileira 26(2):102-104. Department of Preventive Veterinary Medicine and Animal Health, Faculty of Veterinary Medicine and Animal Production, University of São Paulo, Av. Prof. Orlando Marques de Paiva 87, São Paulo, SP 05508-900, Brazil. E-mail: danmoura@aptaregional.sp.gov.br The prevalence of anti-Leptospira spp antibodies was investigated in 2,109 female cattle from 86 herds of Monte Negro municipality, Rondônia, Brazil. Sera samples were evaluated by Microscopic Agglutination Test against 24 leptospira serovars. Titers =100 for at least one of 24 leptospira serovars were detected in 1,114 cows (52.8%) from 82 (95.3%) herds. The adjusted overall prevalence for Monte Negro municipality was 53.9% (49-58.7%; CI: 95%). The most prevalent serovars were Hardjo (14.5%), Wolffi (12.3%), Shermani (10.8%), Patoc (7.9%), and Hebdomadis (6.1%). Other serovars worldwidely reported like Bratislava, Pomona and Grippotyphosa were detected in low levels.

Abstract in English:

Caron L., Brum M.C.S., Moraes M.P., Golde W.T., Arns C.W. & Grubman M.J. 2005. Granulocyte-macrophage colony-stimulating factor does not increase the potency or efficacy of a foot-and-mouth disease virus subunit vaccine. Pesquisa Veterinária Brasileira 25(3):150-158. USDA, ARS, PIADC-FMD Research Unit, PO.Box 848, Greenport, NY 11944 0848, USA. E-mail: mgrubman@piadc.ars.usda.gov Foot-and-mouth disease (FMD) is one of the most feared diseases of livestock worldwide. Vaccination has been a very effective weapon in controlling the disease, however a number of concerns with the current vaccine including the inability of approved diagnostic tests to reliably distinguish vaccinated from infected animals and the need for high containment facilities for vaccine production, have limited its use during outbreaks in countries previously free of the disease. A number of FMD vaccine candidates have been tested and a replication-defective human adenovirus type 5 (Ad5) vector containing the FMDV capsid (P1-2A) and 3C protease coding regions has been shown to completely protect pigs against challenge with the homologous virus (FMDV A12 and A24). An Ad5-P1-2A+3C vaccine for FMDV O1 Campos (Ad5-O1C), however, only induced a low FMDV-specific neutralizing antibody response in swine potency tests. Granulocyte-macrophage colony-stimulating factor (GM-CSF) has been successfully used to stimulate the immune response in vaccine formulations against a number of diseases, including HIV, hepatitis C and B. To attempt to improve the FMDV-specific immune response induced by Ad5-O1C, we inoculated swine with Ad5-O1C and an Ad5 vector containing the gene for porcine GM-CSF (pGM-CSF). However, in the conditions used in this trial, pGM-CSF did not improve the immune response to Ad5-O1C and adversely affected the level of protection of swine challenged with homologous FMDV.

• Foot-and-mouth disease virus O1 Campos Adenovirus Granulocyte-macrophage colony-stimulating factor.

Abstract in Portuguese:

Caron L., Brum M.C.S., Moraes M.P., Golde W.T., Arns C.W. & Grubman M.J. 2005. Granulocyte-macrophage colony-stimulating factor does not increase the potency or efficacy of a foot-and-mouth disease virus subunit vaccine. Pesquisa Veterinária Brasileira 25(3):150-158. USDA, ARS, PIADC-FMD Research Unit, PO.Box 848, Greenport, NY 11944 0848, USA. E-mail: mgrubman@piadc.ars.usda.gov Foot-and-mouth disease (FMD) is one of the most feared diseases of livestock worldwide. Vaccination has been a very effective weapon in controlling the disease, however a number of concerns with the current vaccine including the inability of approved diagnostic tests to reliably distinguish vaccinated from infected animals and the need for high containment facilities for vaccine production, have limited its use during outbreaks in countries previously free of the disease. A number of FMD vaccine candidates have been tested and a replication-defective human adenovirus type 5 (Ad5) vector containing the FMDV capsid (P1-2A) and 3C protease coding regions has been shown to completely protect pigs against challenge with the homologous virus (FMDV A12 and A24). An Ad5-P1-2A+3C vaccine for FMDV O1 Campos (Ad5-O1C), however, only induced a low FMDV-specific neutralizing antibody response in swine potency tests. Granulocyte-macrophage colony-stimulating factor (GM-CSF) has been successfully used to stimulate the immune response in vaccine formulations against a number of diseases, including HIV, hepatitis C and B. To attempt to improve the FMDV-specific immune response induced by Ad5-O1C, we inoculated swine with Ad5-O1C and an Ad5 vector containing the gene for porcine GM-CSF (pGM-CSF). However, in the conditions used in this trial, pGM-CSF did not improve the immune response to Ad5-O1C and adversely affected the level of protection of swine challenged with homologous FMDV.

Abstract in English:

Beltrão N., Furian T.Q., Leão J.A., Pereira R.A., Moraes L.B. & Canal C.W. 2004. [Detection of infectious laryngotracheitis virus in chickens in Brazil.] Detecção do vírus da laringotraqueíte das galinhas no Brasil. Pesquisa Veterinária Brasileira 24(2):85-88. Centro de Diagnóstico e Pesquisa em Patologia Aviária (CDPA), Faculdade de Veterinária, UFRGS, Porto Alegre, RS 91540-000, Brazil. E-mail: nilzaneb@hotmail.com A study was carried out in search for evidences of infectious laryngotracheitis virus (ILTV) infections in some Brazilian chicken flocks. Tracheal tissues and swabs were collected from 10 different flocks of layers and broilers displaying respiratory signs of disease. Samples were processes for virus isolation in embryonated eggs and the membranes examined by histopathology. In addition, specimens were examined by polymerase chain reaction (PCR). Three flocks had ILTV positive chickens by virus isolation and PCR. These results confirm the occurrence of ILTV in chickens in Brazil.

• Infectious laryngotracheitis virus PCR virus isolation avian pathology.

Abstract in Portuguese:

Beltrão N., Furian T.Q., Leão J.A., Pereira R.A., Moraes L.B. & Canal C.W. 2004. [Detection of infectious laryngotracheitis virus in chickens in Brazil.] Detecção do vírus da laringotraqueíte das galinhas no Brasil. Pesquisa Veterinária Brasileira 24(2):85-88. Centro de Diagnóstico e Pesquisa em Patologia Aviária (CDPA), Faculdade de Veterinária, UFRGS, Porto Alegre, RS 91540-000, Brazil. E-mail: nilzaneb@hotmail.com A study was carried out in search for evidences of infectious laryngotracheitis virus (ILTV) infections in some Brazilian chicken flocks. Tracheal tissues and swabs were collected from 10 different flocks of layers and broilers displaying respiratory signs of disease. Samples were processes for virus isolation in embryonated eggs and the membranes examined by histopathology. In addition, specimens were examined by polymerase chain reaction (PCR). Three flocks had ILTV positive chickens by virus isolation and PCR. These results confirm the occurrence of ILTV in chickens in Brazil.

Abstract in English:

Canal C.W., Ferreira D.J., Macagnan M., Fallavena L.C.B., Moraes H.L.S. & Wald V.B. 2004. Prevalence of antibodies against chicken anaemia virus (CAV) in broiler breeders in Southern Brazil. Pesquisa Veterinária Brasileira 24(2):89-92. Centro de Diagnóstico e Pesquisa em Patologia Aviária (CDPA), Faculdade de Veterinária da Universidade Federal do Rio Grande do Sul, Porto Alegre, RS 91540-000, Brazil. E-mail: claudio.canal@ufrgs.br Chicks infected during the first two weeks of life with chicken anaemia virus (CAV) manifest clinical disease that can be avoided if the breeder hens transfer enough antibodies to their progeny. The objective of the present work was to establish the prevalence and titer of anti-CAV antibodies in some Brazilian broiler hen breeder flocks and verify in which phase of life the birds were infected. A total of 1,709 serum samples from 12 broiler hen flocks vaccinated against CAV and 64 unvaccinated flocks were analyzed for CAV antibodies with an enzyme-linked immunosorbent assay (ELISA). All non-vaccinated breeder flocks were found to be infected with CAV, with 89% of the hens tested presenting antibodies, 52% of these with titers considered high enough to protect their progeny against CAV infection. Likewise, all vaccinated hens had antibody titer to CAV capable of conferring protection to their progeny. Thus, vaccination of hens seems capable of conferring protection to chicks against clinically apparent CAV-associated disease.

• Antibodies avian pathology chicken anemia virus CAV epidemiology prevalence.

Abstract in Portuguese:

Canal C.W., Ferreira D.J., Macagnan M., Fallavena L.C.B., Moraes H.L.S. & Wald V.B. 2004. Prevalence of antibodies against chicken anaemia virus (CAV) in broiler breeders in Southern Brazil. Pesquisa Veterinária Brasileira 24(2):89-92. Centro de Diagnóstico e Pesquisa em Patologia Aviária (CDPA), Faculdade de Veterinária da Universidade Federal do Rio Grande do Sul, Porto Alegre, RS 91540-000, Brazil. E-mail: claudio.canal@ufrgs.br Chicks infected during the first two weeks of life with chicken anaemia virus (CAV) manifest clinical disease that can be avoided if the breeder hens transfer enough antibodies to their progeny. The objective of the present work was to establish the prevalence and titer of anti-CAV antibodies in some Brazilian broiler hen breeder flocks and verify in which phase of life the birds were infected. A total of 1,709 serum samples from 12 broiler hen flocks vaccinated against CAV and 64 unvaccinated flocks were analyzed for CAV antibodies with an enzyme-linked immunosorbent assay (ELISA). All non-vaccinated breeder flocks were found to be infected with CAV, with 89% of the hens tested presenting antibodies, 52% of these with titers considered high enough to protect their progeny against CAV infection. Likewise, all vaccinated hens had antibody titer to CAV capable of conferring protection to their progeny. Thus, vaccination of hens seems capable of conferring protection to chicks against clinically apparent CAV-associated disease.

Abstract in English:

ABSTRACT.- Flores E.F., Weiblen R., Botton S.A., Medeiros M., lrigoyen L.F., Driemeier D., Schuch L.F. & Moraes M.S. 2000. A retrospective search for bovine respiratory syncytial virus (BRSV) antigens in histological specimens by immunofluorescence and immunohistochemistry. [Pesquisa retrospectiva de antígenos do Vírus Respiratório Sincicial Bovino (BRSV) em cortes histológicos por imunofluorescência e imunohistoquímica.] Pesquisa Veterinária Brasileira 20(4): 139-143. Depto Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. Bovine respiratory syncytial vírus (BRSV) has been only sporadically identified as a causa tive agent of respiratory disease in Brazil. This contrasts with frequent reports of clinical and histopathological findings suggestive of BRSV-associated disease. In order to examine a possible involvement of BRSV in cases of calf pneumonia, a retrospective search was performed for BRSV antigens in histological specimens submitted to veterinary diagnostic services from the states of Rio Grande do Sul and Minas Gerais. Ten out of 41 cases examined (24.4%) were positive for BRSV antigens by immunohistochemistry (IPX). Eight of these cases (19.5%) were also positive by indirect immunofluorescence (IFA), and 31 cases (75.6%) were negative in both assays. In the lungs, BRSV antigens were predominantly observed in epithelial cells of bronchioles and less frequently found in alveoli. In one case, antigens were detected only in the epithelium of the alveolar septae. The presence of antigen-positive cells was largely restricted to epithelial cells of these airways. In two cases, positive staining was also observed in cells and cellular debris in the exudate within the pulmonary airways. The clinical cases positive for BRSV antigens were observed mainly in young animals (2 to 12 month-old) from dairy herds. The main microscopic changes included bronchointerstitial pneumonia characterized by thickening of alveolar septae adjacent to airways by mononuclear cell infiltrates, and the presence of alveolar syncytial giant cells. In summary, the results demonstrate the suitability of the immunodetection of viral antigens in routinely fixed tissue specimens as a diagnostic tool for BRSV infection. Moreover, the findings provide further evidence of the importance of BRSVas a respiratory pathogen of young cattle in southeastern and southern Brazil.

• Bovine respiratory syncytial virus BRSV immunohistochemistry immunofluorescence diagnosis Vírus respiratório sincicial bovino imunohistoquímica imunofluorescência diagnóstico.

Abstract in Portuguese:

RESUMO.- Flores E.F., Weiblen R., Botton S.A., Medeiros M., lrigoyen L.F., Driemeier D., Schuch L.F. & Moraes M.S. 2000. A retrospective search for bovine respiratory syncytial virus (BRSV) antigens in histological specimens by immunofluorescence and immunohistochemistry. [Pesquisa retrospectiva de antígenos do Vírus Respiratório Sincicial Bovino (BRSV) em cortes histológicos por imunofluorescência e imunohistoquímica.] Pesquisa Veterinária Brasileira 20(4): 139-143. Depto Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. O Vírus Respiratório Sincicial Bovino (BRSV) tem sido raramente identificado como agente etiológico de doença respiratória em bovinos no Brasil. Isso contrasta com freqüentes relatos clínicos e histopatológicos sugestivos de enfermidade associada ao BRSV. Com o objetivo de investigar o possível envolvimento do BRSV em casos de pneumonia em bovinos jovens, realizou-se uma pesquisa retrospectiva para antígenos do BRSV em cortes histológicos de materiais submetidos a quatro serviços de diagnóstico nos Estados do Rio Grande do Sul e Minas Gerais. Dez entre 41 casos examinados (24,4%) foram positivos para antígenos do BRSV por imunohistoquímica (lPX). Oito desses casos (19,5%) também foram positivos por imunofluorescência (lFA) e 31 casos (75,6%) foram negativos em ambos os testes. Nos pulmões, antígenos virais foram detectados predominantemente nas células epiteliais dos bronquíolos e menos freqüentemente nos alveólos. Em um caso, antígenos virais foram detectados exclusivamente no epitélio alveolar. A presença de células positivas restringiu-se ao epitélio dessas vias aéreas. Em dois casos, antígenos virais foram detectados em células descamativas e em restos celulares no exsudato das vias aéreas. Os casos positivos para antígenos do BRSV eram oriundos principalmente de animais jovens (2 a 12 meses de idade) de rebanhos leiteiros. As principais alterações histológicas observadas foram pneumonia bronco-intersticial caracterizada por espessamento dos septos alveolares adjacentes às vias aéreas, infiltrados de células mononucleares e presença de células gigantes multinucleadas nos alveólos. Os resultados obtidos demonstram a utilidade dos métodos de detecção de antígenos em cortes histológicos para o diagnóstico da infecção pelo BRSV. Além disso, esses resultados confirmam evidências anteriores sobre a importância do BRSV como patógeno respiratório no sudeste e sul do Brasil.