PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

Simionatto S., Vaz E.K., Michelon A., Seixas F.K., Dellagostin O.A. 2005. [Development and evaluation of new strategies for immunization against swine colibacillosis.] Desenvolvimento e avaliação de novas estratégias de imunização contra colibacilose suína. Pes-quisa Veterinária Brasileira 25(2):84-90. Laboratório de Biologia Molecular, Centro de Bio-tecnologia, UFPel, Campus Capão do Leão, Cx. Postal 354, Pelotas, RS 96010-900, Brazil. E-mail: ssimionatto@bol.com.br Swine colibacillosis caused by enterotoxigenic Escherichia coli remains one of the main sanitary problems in pig farms. The recombinant DNA technology offers the possibility of developing new immunization strategies. This paper describes the development of a subunit vaccine through the expression and purification of the E. coli K88 FaeC fimbrial protein. The gene that codes for this antigen was amplified by PCR and cloned into an E. coli expression vector fused to a 6X histidine tag. The recombinant protein was purified by affinity chromatography and used for mice immunization. In parallel, the same gene was cloned into an eucariotic expression vector with the addition of the Kozak sequence for improving translation of this gene in muscle cells. The resulting plasmid named pUP310 was purified in large scale and used to immunize mice. The immune response afforded by both forms of immunization was monitored by ELISA. There was an immune response in mice inoculated with pUP310 and purified FaeC. It was possible to detect anti-FaeC antibodies 42 days after the first inoculation. The antibody titer increased with time, being still detectable 7 months after the first inoculation. It is concluded that recombinant FaeC and pUP310 are potential tools for immunization of swine against E. coli K88.

• Swine colibacillosis FaeC Escherichia coli DNA vaccine recombinant vaccine.

Abstract in Portuguese:

Simionatto S., Vaz E.K., Michelon A., Seixas F.K., Dellagostin O.A. 2005. [Development and evaluation of new strategies for immunization against swine colibacillosis.] Desenvolvimento e avaliação de novas estratégias de imunização contra colibacilose suína. Pes-quisa Veterinária Brasileira 25(2):84-90. Laboratório de Biologia Molecular, Centro de Bio-tecnologia, UFPel, Campus Capão do Leão, Cx. Postal 354, Pelotas, RS 96010-900, Brazil. E-mail: ssimionatto@bol.com.br Swine colibacillosis caused by enterotoxigenic Escherichia coli remains one of the main sanitary problems in pig farms. The recombinant DNA technology offers the possibility of developing new immunization strategies. This paper describes the development of a subunit vaccine through the expression and purification of the E. coli K88 FaeC fimbrial protein. The gene that codes for this antigen was amplified by PCR and cloned into an E. coli expression vector fused to a 6X histidine tag. The recombinant protein was purified by affinity chromatography and used for mice immunization. In parallel, the same gene was cloned into an eucariotic expression vector with the addition of the Kozak sequence for improving translation of this gene in muscle cells. The resulting plasmid named pUP310 was purified in large scale and used to immunize mice. The immune response afforded by both forms of immunization was monitored by ELISA. There was an immune response in mice inoculated with pUP310 and purified FaeC. It was possible to detect anti-FaeC antibodies 42 days after the first inoculation. The antibody titer increased with time, being still detectable 7 months after the first inoculation. It is concluded that recombinant FaeC and pUP310 are potential tools for immunization of swine against E. coli K88.

Abstract in English:

França T.N., Ribeiro C.T., Cunha B.M. & Peixoto P.V. 2005. [Porcine Circovirosis: a review.] Circovirose Suína. Pesquisa Veterinária Brasileira 25(2):59-72. Universidade Estácio de Sá, Curso de Medicina Veterinária, Disciplina de Anatomia Patológica, Estrada Boca do Mato 850, Vargem Pequena, RJ 22783-320, Brazil. E-mail: ticianaf@uol.com.br The literature of Porcine Circovirosis, including the main data on epidemiology and clinical, macroscopic and microscopic alterations of the infection of swine by Porcine Circovirus type 2 (PCV-2), is reviewed. There are various forms of infection: the [Porcine] Postweaning Multisystemic Wasting Syndrome (PMWS), Porcine Congenital Tremor, Porcine Dermatitis and Nephropathy Syndrome, and other associated or correlated diseases as the Porcine Reproductive and Respiratory Syndrome, Proliferative Necrotizing Pneumonia, and reproductive disorders. As PMWS already has been reported from southern Brazil and from the state of Rio de Janeiro, the objective of this review is to draw attention to the implications of this virosis for swine production in Brazil and its economical importance.

• Porcine Circovirosis Porcine Circovirus type 2 Porcine Postweaning Multisystemic Wasting Syndrome Porcine Congenital Tremor Porcine Dermatitis and Nephropathy Syndrome Proliferative Necrotizing Pneumonia reproductive disorders of swine.

Abstract in Portuguese:

França T.N., Ribeiro C.T., Cunha B.M. & Peixoto P.V. 2005. [Porcine Circovirosis: a review.] Circovirose Suína. Pesquisa Veterinária Brasileira 25(2):59-72. Universidade Estácio de Sá, Curso de Medicina Veterinária, Disciplina de Anatomia Patológica, Estrada Boca do Mato 850, Vargem Pequena, RJ 22783-320, Brazil. E-mail: ticianaf@uol.com.br The literature of Porcine Circovirosis, including the main data on epidemiology and clinical, macroscopic and microscopic alterations of the infection of swine by Porcine Circovirus type 2 (PCV-2), is reviewed. There are various forms of infection: the [Porcine] Postweaning Multisystemic Wasting Syndrome (PMWS), Porcine Congenital Tremor, Porcine Dermatitis and Nephropathy Syndrome, and other associated or correlated diseases as the Porcine Reproductive and Respiratory Syndrome, Proliferative Necrotizing Pneumonia, and reproductive disorders. As PMWS already has been reported from southern Brazil and from the state of Rio de Janeiro, the objective of this review is to draw attention to the implications of this virosis for swine production in Brazil and its economical importance.

Abstract in English:

Hofer E. & Reis C.M.F. 2005. [Species and serovars of Listeria isolated from sick and clinically healthy animals in Brazil.] Espécies e sorovares de Listeria isolados de animais doentes e portadores no Brasil. Pesquisa Veterinária Brasileira 25(2):79-83. Laboratório de Zoonoses Bacterianas, Depto Bacteriologia, Instituto Oswaldo Cruz/FIOCRUZ, Rio de Janeiro, RJ 21045-900, Brazil. E-mail: ehofer@ioc.fiocruz.br Two hundred fourty-six strains of the genus Listeria were isolated from sick and clinically healthy animals, collected in three different regions of Brazil during 1971-2000. About 88.2% (217 cultures) yielded Listeria species from faecal specimens of healthy cattle and 29 strains (11.7%) were isolated from sick animals: 15 (6.0%) from central nervous system (CNS) and 14(5.6%) were from otherwise sterile sites. Phenotyping techniques were used to characterize the Listeria isolates. The commonest were L. innocua 6a and non-typable (140/56.9%), L. monocytogenes 4a (37/15.0%) and 4b (22/8.9%), originated mainly from stools of healthy cattle. From sick animals the predominant species and serovars were L. monocytogenes 4b (14/5.6%), and the higher incidence was observed in ruminants (12/4.8%) and 8/3.2% of the serovar 1a from other animal species (rodents and canines) mainly isolated from CNS samples.

• Listeria species serovars sick and healthy animals.

Abstract in Portuguese:

Hofer E. & Reis C.M.F. 2005. [Species and serovars of Listeria isolated from sick and clinically healthy animals in Brazil.] Espécies e sorovares de Listeria isolados de animais doentes e portadores no Brasil. Pesquisa Veterinária Brasileira 25(2):79-83. Laboratório de Zoonoses Bacterianas, Depto Bacteriologia, Instituto Oswaldo Cruz/FIOCRUZ, Rio de Janeiro, RJ 21045-900, Brazil. E-mail: ehofer@ioc.fiocruz.br Two hundred fourty-six strains of the genus Listeria were isolated from sick and clinically healthy animals, collected in three different regions of Brazil during 1971-2000. About 88.2% (217 cultures) yielded Listeria species from faecal specimens of healthy cattle and 29 strains (11.7%) were isolated from sick animals: 15 (6.0%) from central nervous system (CNS) and 14(5.6%) were from otherwise sterile sites. Phenotyping techniques were used to characterize the Listeria isolates. The commonest were L. innocua 6a and non-typable (140/56.9%), L. monocytogenes 4a (37/15.0%) and 4b (22/8.9%), originated mainly from stools of healthy cattle. From sick animals the predominant species and serovars were L. monocytogenes 4b (14/5.6%), and the higher incidence was observed in ruminants (12/4.8%) and 8/3.2% of the serovar 1a from other animal species (rodents and canines) mainly isolated from CNS samples.

Abstract in English:

Penatti M.P.A., Silva A.S., Valadares G.F. & Leite D.S. 2005. Occurrence of F42 colonization factor in Escherichia coli strains isolated from piglets with diarrhea. Pesquisa Veterinária Brasileira 25(1):31-33. Depto Microbiologia e Imunologia, Instituto de Biologia, Unicamp, Campinas, SP 13081-970, Brazil. E-mail: domingos@unicamp.br The objective of this study was to determine the presence of the colonization factor F42 in 168 strains of Escherichia coli isolated from diarrheic stools of newborn piglets. The presence of F42 in 12 (7.1%) strains was detected with the agglutination test. Through the Polymerase Chain Reaction (PCR) of F42 positive strains, gene encoding enterotoxins (ST-I, ST-II, LT-I and LT-II) were detected. The finding of ST-I/ST-II genes in 50% of the strains, ST-I (16%) and ST-II (25%) indicates a strong association of FC F42 with heat-stable enterotoxins (91%). In contrast, the thermolabile enterotoxin (LT-I and LT-II) genes were not detected. Serogroups of F42 positive strains were determined, serogroup O8 being the most prevalent (41,7%). Other serogroups, as there are O9, O11, O18, O32, O35, O98 and O101, were also identified. Thus, FC F42 was confirmed as an additional factor of virulence in the pathogenesis of porcine colibacillosis.

• Escherichia coli F42 piglets fimbriae diarrhea PCR.

Abstract in Portuguese:

Penatti M.P.A., Silva A.S., Valadares G.F. & Leite D.S. 2005. Occurrence of F42 colonization factor in Escherichia coli strains isolated from piglets with diarrhea. Pesquisa Veterinária Brasileira 25(1):31-33. Depto Microbiologia e Imunologia, Instituto de Biologia, Unicamp, Campinas, SP 13081-970, Brazil. E-mail: domingos@unicamp.br The objective of this study was to determine the presence of the colonization factor F42 in 168 strains of Escherichia coli isolated from diarrheic stools of newborn piglets. The presence of F42 in 12 (7.1%) strains was detected with the agglutination test. Through the Polymerase Chain Reaction (PCR) of F42 positive strains, gene encoding enterotoxins (ST-I, ST-II, LT-I and LT-II) were detected. The finding of ST-I/ST-II genes in 50% of the strains, ST-I (16%) and ST-II (25%) indicates a strong association of FC F42 with heat-stable enterotoxins (91%). In contrast, the thermolabile enterotoxin (LT-I and LT-II) genes were not detected. Serogroups of F42 positive strains were determined, serogroup O8 being the most prevalent (41,7%). Other serogroups, as there are O9, O11, O18, O32, O35, O98 and O101, were also identified. Thus, FC F42 was confirmed as an additional factor of virulence in the pathogenesis of porcine colibacillosis.

Abstract in English:

Assis R.A., Lobato F.C.F., Serakides R., Santos R.L., Dias G.R.C., Nascimento R.A.P., Abreu V.L.V, Parreiras P.M. & Uzal F.A. 2005. Immunohistochemical detection of Clostridia species in paraffin-embedded tissues of experimentally inoculated guinea pigs. Pesquisa Veterinária Brasileira 25(1):4-8. Departamento de Medicina Veterinária Preventiva, Escola de Veterinária, Universidade Federal de Minas Gerais, Av. Presidente Antônio Carlos 6627, Cx. Postal 567, Belo Horizonte, MG 30123-970, Brazil. E-mail: assisra@rwnet.com.br Blackleg is caused by Clostridium chauvoei, whereas malignant oedema is caused by C. chauvoei, C. septicum, C. sordellii, C. perfringens type A, and/or C. novyi type A. Anti-C. chauvoei, anti-C. septicum, anti-C. sordellii and anti-C. novyi type A polyclonal antibodies were produced in rabbits and purified in a column of DEAE-cellulose. Aliquots of the antisera were conjugated with fluorescein isothiocyanate and the remaining was used for the streptavidin biotin peroxidase technique (SBPT). SBPT was standardized to detect C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs. SBPT was compared to a fluorescent antibody technique (FAT). Sections and smears of muscle from inoculation area (MIA), heart, liver, spleen and kidney, were obtained for both SBPT and FAT. Cross-reactions between the different Clostridial species were not observed. C. chauvoei and C. septicum were detected in all specimens from the animals inoculated with these microorganisms, while only sections of muscle obtained from all the animals inoculated with C. sordellii and C. novyi type A were positive. The same results observed by the SBPT, were obtained on tissue smears of these microorganisms stained by the FAT. The results indicate that SBPT is suitable for detection of C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs.

• Blackleg malignant oedema Clostridia guinea pigs fluorescent antibody technique streptavidin biotin peroxidase technique.

Abstract in Portuguese:

Assis R.A., Lobato F.C.F., Serakides R., Santos R.L., Dias G.R.C., Nascimento R.A.P., Abreu V.L.V, Parreiras P.M. & Uzal F.A. 2005. Immunohistochemical detection of Clostridia species in paraffin-embedded tissues of experimentally inoculated guinea pigs. Pesquisa Veterinária Brasileira 25(1):4-8. Departamento de Medicina Veterinária Preventiva, Escola de Veterinária, Universidade Federal de Minas Gerais, Av. Presidente Antônio Carlos 6627, Cx. Postal 567, Belo Horizonte, MG 30123-970, Brazil. E-mail: assisra@rwnet.com.br Blackleg is caused by Clostridium chauvoei, whereas malignant oedema is caused by C. chauvoei, C. septicum, C. sordellii, C. perfringens type A, and/or C. novyi type A. Anti-C. chauvoei, anti-C. septicum, anti-C. sordellii and anti-C. novyi type A polyclonal antibodies were produced in rabbits and purified in a column of DEAE-cellulose. Aliquots of the antisera were conjugated with fluorescein isothiocyanate and the remaining was used for the streptavidin biotin peroxidase technique (SBPT). SBPT was standardized to detect C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs. SBPT was compared to a fluorescent antibody technique (FAT). Sections and smears of muscle from inoculation area (MIA), heart, liver, spleen and kidney, were obtained for both SBPT and FAT. Cross-reactions between the different Clostridial species were not observed. C. chauvoei and C. septicum were detected in all specimens from the animals inoculated with these microorganisms, while only sections of muscle obtained from all the animals inoculated with C. sordellii and C. novyi type A were positive. The same results observed by the SBPT, were obtained on tissue smears of these microorganisms stained by the FAT. The results indicate that SBPT is suitable for detection of C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs.

Abstract in English:

Seitz A.L., Colodel E.M., Barros S.S. & Driemeier D. 2005. [Experimental poisoning by Sida carpinifolia (Malvaceae) in sheep.] Intoxicação experimental por Sida carpinifolia (Malvaceae) em ovinos. Pesquisa Veterinária Brasileira 25(1):15-20. Departamento de Patologia Clínica Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS 91540-000, Brazil. E-mail: alseitz@terra.com.br. Seven sheep received dry crushed Sida carpinifolia L.f. One of them died at 18 and other at 53 days of the experiment. Four others were euthanatized and necropsied at 30, 45, 75 and 100 days. For one sheep the supply of S. carpinifolia was interrupted on the 80th day of the experiment, and 70 days later the animal was euthanized and necropsied. The minimal amount of the dry plant consumed was 11 g/kg and the maximum was 30 g/kg. The progression of clinical findings was similar in six animals with slight diarrhea at 20 days of experiment. Neurological signs were observed at 25 days and included ataxia with dysmetria, muscle tremors of the head, atypical postural reactions, frequent falls, sluggish of movements, difficulty in grazing and swallowing. These signs were enhanced when the animals were forced to walk. Four of the animals presented progressive emaciation. The sheep whose supply of the plant was interrupted recovered gradually, and 11 days after the animal returned to normal. During necropsy, only enlarged mesenteric lymph nodes were observed. The histological alterations were more significant in the central nervous system, with multiple and severe cytoplasmic distention and vacuolation which affects specially Purkinje cells of the cerebellum, neurons of cerebral cortex, thalamus, midbrain and the ventral horn of spinal cord. Axonal spheroids in the brain, more frequently in the granular layer of cerebellum were also observed. The cytoplasmic vacuolation was also found in pancreatic acinar cells, renal tubules, thyroid follicular epithelium, hepatocytes and macrophages of lymphoid organs. The ultrastructural lesions observed were cytoplasmic vacuolation, some surrounded by membranes in Purkinje cells of cerebellum and thyroid follicular cells. The sheep, which had S. carpinifolia withdrawn from its diet for 70 days, had no significant histological alterations.

• Toxic plants plant poisoning Sida carpinifolia Malvaceae lysosomal storage disease swainsonine electronic microscopy.

Abstract in Portuguese:

Seitz A.L., Colodel E.M., Barros S.S. & Driemeier D. 2005. [Experimental poisoning by Sida carpinifolia (Malvaceae) in sheep.] Intoxicação experimental por Sida carpinifolia (Malvaceae) em ovinos. Pesquisa Veterinária Brasileira 25(1):15-20. Departamento de Patologia Clínica Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS 91540-000, Brazil. E-mail: alseitz@terra.com.br. Seven sheep received dry crushed Sida carpinifolia L.f. One of them died at 18 and other at 53 days of the experiment. Four others were euthanatized and necropsied at 30, 45, 75 and 100 days. For one sheep the supply of S. carpinifolia was interrupted on the 80th day of the experiment, and 70 days later the animal was euthanized and necropsied. The minimal amount of the dry plant consumed was 11 g/kg and the maximum was 30 g/kg. The progression of clinical findings was similar in six animals with slight diarrhea at 20 days of experiment. Neurological signs were observed at 25 days and included ataxia with dysmetria, muscle tremors of the head, atypical postural reactions, frequent falls, sluggish of movements, difficulty in grazing and swallowing. These signs were enhanced when the animals were forced to walk. Four of the animals presented progressive emaciation. The sheep whose supply of the plant was interrupted recovered gradually, and 11 days after the animal returned to normal. During necropsy, only enlarged mesenteric lymph nodes were observed. The histological alterations were more significant in the central nervous system, with multiple and severe cytoplasmic distention and vacuolation which affects specially Purkinje cells of the cerebellum, neurons of cerebral cortex, thalamus, midbrain and the ventral horn of spinal cord. Axonal spheroids in the brain, more frequently in the granular layer of cerebellum were also observed. The cytoplasmic vacuolation was also found in pancreatic acinar cells, renal tubules, thyroid follicular epithelium, hepatocytes and macrophages of lymphoid organs. The ultrastructural lesions observed were cytoplasmic vacuolation, some surrounded by membranes in Purkinje cells of cerebellum and thyroid follicular cells. The sheep, which had S. carpinifolia withdrawn from its diet for 70 days, had no significant histological alterations.

Abstract in English:

França T.N., Peixoto P.V., Brito M.F., Driemeier D., Mores N. & Zanella J. 2005. [Outbreak of Circovirosis (Porcine Postweaning Multisystemic Wasting Syndrome) in the state of Rio de Janeiro, Brazil.] Surto de Circovirose (Síndrome Definhante Multissistêmica de Suínos Desmamados) no estado do Rio de Janeiro. Pesquisa Veterinária Brasileira 25(1):39-53. Universidade Estácio de Sá, Curso de Medicina Veterinária, Disciplina de Anatomia Patológica, Estrada Boca do Mato 850, Vargem Pequena, RJ 22783-320, Brazil. E-mail: ticianaf@uol.com.br The first outbreak of Postweaning Multisystemic Wasting Syndrome (PMWS) in swine, which occurred in southeastern Brazil, in the state of Rio de Janeiro, is described. The disease, which affects mainly weaned about 4 month-old pigs, caused the death of at least 14 animals. The property, where the outbreak occurred, had inadequate sanitary and management conditions. Clinically the disease was characterized by wasting, poor development, cough, tachypnoea, dispnoea, diarrhoea, ataxia, tremors after stimulation, decubitus and convulsions. The course of the disease was acute or subacute. The most important post-mortem findings were enlarged lymphnodes, non-collapsed lungs, with consolidated areas mainly in the cranial lobes. Histological lesions consisted mainly of lymphohistiocytic infiltration with multinucleate giant cells in lymph nodes, spleen, Peyer’s patches, kidney, lung and liver, depletion or lymphoid hyperplasia, as well as lymphohistiocytic interstitial pneumonia and areas of secondary bronchopneumonia. The diagnosis was established through observations of the symptoms and typical lesions, and was confirmed by immunohistochemical examination and PCR. The objective of this study was to characterize the epidemiological, clinical and pathological aspects of the outbreak of PMWS, because of the severe direct or indirect economical losses caused by the disease to the world pig industry.

• Postweaning multisystemic wasting syndrome porcine circovirus type 2 pathology immunohistochemistry PCR.

Abstract in Portuguese:

França T.N., Peixoto P.V., Brito M.F., Driemeier D., Mores N. & Zanella J. 2005. [Outbreak of Circovirosis (Porcine Postweaning Multisystemic Wasting Syndrome) in the state of Rio de Janeiro, Brazil.] Surto de Circovirose (Síndrome Definhante Multissistêmica de Suínos Desmamados) no estado do Rio de Janeiro. Pesquisa Veterinária Brasileira 25(1):39-53. Universidade Estácio de Sá, Curso de Medicina Veterinária, Disciplina de Anatomia Patológica, Estrada Boca do Mato 850, Vargem Pequena, RJ 22783-320, Brazil. E-mail: ticianaf@uol.com.br The first outbreak of Postweaning Multisystemic Wasting Syndrome (PMWS) in swine, which occurred in southeastern Brazil, in the state of Rio de Janeiro, is described. The disease, which affects mainly weaned about 4 month-old pigs, caused the death of at least 14 animals. The property, where the outbreak occurred, had inadequate sanitary and management conditions. Clinically the disease was characterized by wasting, poor development, cough, tachypnoea, dispnoea, diarrhoea, ataxia, tremors after stimulation, decubitus and convulsions. The course of the disease was acute or subacute. The most important post-mortem findings were enlarged lymphnodes, non-collapsed lungs, with consolidated areas mainly in the cranial lobes. Histological lesions consisted mainly of lymphohistiocytic infiltration with multinucleate giant cells in lymph nodes, spleen, Peyer’s patches, kidney, lung and liver, depletion or lymphoid hyperplasia, as well as lymphohistiocytic interstitial pneumonia and areas of secondary bronchopneumonia. The diagnosis was established through observations of the symptoms and typical lesions, and was confirmed by immunohistochemical examination and PCR. The objective of this study was to characterize the epidemiological, clinical and pathological aspects of the outbreak of PMWS, because of the severe direct or indirect economical losses caused by the disease to the world pig industry.

Abstract in English:

Spilki F.R., Silva A.D., Batista H.B.C.R., Oliveira A.P., Winkelmann E., Franco A.C., Porciúncula J.A. & Roehe P.M. 2005. Field evaluation of safety during gestation and horizontal spread of a recombinant differential bovine herpesvirus 1 (BoHV-1) vaccine. Pesquisa Veterinária Brasileira 25(1):54-58. Instituto de Pesquisa Veterinária Desidério Finamor, Fepagro-Saúde Animal, Cx. Postal 47, Eldorado do Sul, RS 92990-000, Brazil. E-mail: proehe@ufrgs.br Bovine herpesvirus type 1 (BoHV-1) is recognized as a major cause of respiratory, reproductive disease and abortion in cattle. Vaccination is widely applied to minimize losses induced by BoHV-1 infections; however, vaccination of dams during pregnancy with modified live virus (MLV) vaccines has been occasionally associated to abortions. We have previously reported the development of a BoHV-1 recombinant virus, constructed with basis on a Brazilian BoHV-1 (Franco et al. 2002a) from which the gene coding for glycoprotein E (gE) was deleted (gE-) by genetic manipulation. Such recombinant has been previously evaluated in its potential as a differential vaccine (gE- vaccine) that allows differentiation between vaccinated and infected animals. Here, in the first part of the present study, the safety of the gE- vaccine during pregnancy was evaluated by the intramuscular inoculation of 107.4 tissue culture 50 % infective doses (TCID50) of the virus into 22 pregnant dams (14 BoHV-1 seronegative; 8 seropositive), at different stages of gestation. Other 15 pregnant dams were kept as non-vaccinated controls. No abortions, stillbirths or fetal abnormalities were seen after vaccination. Seroconversion was observed in both groups of previously seronegative vaccinated animals. In the second part of the study, the potential of the gE- vaccine virus to spread among beef cattle under field conditions was examined. Four heifers were inoculated intranasally with a larger amount (107,6 TCID50) of the gE- vaccine (to increase chances of transmission) and mixed with other sixteen animals at the same age and body condition, in the same grazing area, at a population density equal to the average cattle farming density within the region (one cattle head per 10,000 m2), for 180 days. All animals were monitored daily for clinical signs. Serum samples were collected on days 0, 30, 60 and 180 post-vaccination. Seroconversion was observed only in vaccinated heifers. These results indicate that, under the conditions of the present study, the gE- vaccine virus did not cause any noticeable harmful effect on pregnant dams and on its offspring and did not spread horizontally among cattle.

• Bovine herpesvirus 1 BoHV-1 recombinant gE- vaccine.

Abstract in Portuguese:

Spilki F.R., Silva A.D., Batista H.B.C.R., Oliveira A.P., Winkelmann E., Franco A.C., Porciúncula J.A. & Roehe P.M. 2005. Field evaluation of safety during gestation and horizontal spread of a recombinant differential bovine herpesvirus 1 (BoHV-1) vaccine. Pesquisa Veterinária Brasileira 25(1):54-58. Instituto de Pesquisa Veterinária Desidério Finamor, Fepagro-Saúde Animal, Cx. Postal 47, Eldorado do Sul, RS 92990-000, Brazil. E-mail: proehe@ufrgs.br Bovine herpesvirus type 1 (BoHV-1) is recognized as a major cause of respiratory, reproductive disease and abortion in cattle. Vaccination is widely applied to minimize losses induced by BoHV-1 infections; however, vaccination of dams during pregnancy with modified live virus (MLV) vaccines has been occasionally associated to abortions. We have previously reported the development of a BoHV-1 recombinant virus, constructed with basis on a Brazilian BoHV-1 (Franco et al. 2002a) from which the gene coding for glycoprotein E (gE) was deleted (gE-) by genetic manipulation. Such recombinant has been previously evaluated in its potential as a differential vaccine (gE- vaccine) that allows differentiation between vaccinated and infected animals. Here, in the first part of the present study, the safety of the gE- vaccine during pregnancy was evaluated by the intramuscular inoculation of 107.4 tissue culture 50 % infective doses (TCID50) of the virus into 22 pregnant dams (14 BoHV-1 seronegative; 8 seropositive), at different stages of gestation. Other 15 pregnant dams were kept as non-vaccinated controls. No abortions, stillbirths or fetal abnormalities were seen after vaccination. Seroconversion was observed in both groups of previously seronegative vaccinated animals. In the second part of the study, the potential of the gE- vaccine virus to spread among beef cattle under field conditions was examined. Four heifers were inoculated intranasally with a larger amount (107,6 TCID50) of the gE- vaccine (to increase chances of transmission) and mixed with other sixteen animals at the same age and body condition, in the same grazing area, at a population density equal to the average cattle farming density within the region (one cattle head per 10,000 m2), for 180 days. All animals were monitored daily for clinical signs. Serum samples were collected on days 0, 30, 60 and 180 post-vaccination. Seroconversion was observed only in vaccinated heifers. These results indicate that, under the conditions of the present study, the gE- vaccine virus did not cause any noticeable harmful effect on pregnant dams and on its offspring and did not spread horizontally among cattle.

Abstract in English:

D'Ávila da Silva A., Sortica V.A., Braga A.C., Spilki F.R., Franco A.C., Esteves P.A., Rijsewijk F., Rosa J.C.A., Batista H.B.C.R., Oliveira A.P. & Roehe P.M. 2005. [Antigenic and molecular characterization of eight samples of Aujeszky’s disease virus isolated in the state of Rio Grande do Sul, Brazil, in 2003.] Caracterização antigênica e molecular de oito amostras do virus da doença de Aujeszky isoladas no estado do Rio Grande do Sul em 2003. Pesquisa Veterinária Brasileira 25(1):21-24. Instituto de Pesquisas Veterinárias Desidério Finamor (IPVDF), Fepagro Saúde Animal, Estrada do Conde 6000, Cx. Postal 47, Eldorado do Sul, RS 92990-000, Brazil. E-mail: proehe@ufrgs.br Pseudorabies or Aujeszky’s disease (AD), caused by pseudorabies virus (PRV) is a major concern in swine production. In the state of Rio Grande do Sul, Brazil, AD was only detected in 1954, in cattle. In 2003 two outbreaks of encephalitis occurred on the northern region of the state, close to the border with the state of Santa Catarina. Pseudorabies virus (PRV) was isolated from distinct farms within the region and subjected to antigenic and genomic analyses. These isolates were compared with prototype strains NIA-3 and NP. Antigenic characterization with a panel of monoclonal antibodies (Mabs) directed to viral glycoproteins (gB, gC, gD and gE,) was performed by an imunoperoxidase monolayer assay (IPMA) on infected cell monolayers. Genomic characterization was carried out by restriction enzyme analysis (REA) of the whole DNA viral genome with Bam HI. The antigenic profile of the eight isolates from Rio Grande do Sul as well as strains NIA-3 and NP were similar. REA analysis revealed that all isolates from Rio Grande do Sul displayed a genomic type II arrangement, a genotype often found in other outbreaks of AD previously reported in other Brazilian states. The results obtained suggest that the eight isolates examined here were similar.

• Aujeszky's disease epidemiology.

Abstract in Portuguese:

D'Ávila da Silva A., Sortica V.A., Braga A.C., Spilki F.R., Franco A.C., Esteves P.A., Rijsewijk F., Rosa J.C.A., Batista H.B.C.R., Oliveira A.P. & Roehe P.M. 2005. [Antigenic and molecular characterization of eight samples of Aujeszky’s disease virus isolated in the state of Rio Grande do Sul, Brazil, in 2003.] Caracterização antigênica e molecular de oito amostras do virus da doença de Aujeszky isoladas no estado do Rio Grande do Sul em 2003. Pesquisa Veterinária Brasileira 25(1):21-24. Instituto de Pesquisas Veterinárias Desidério Finamor (IPVDF), Fepagro Saúde Animal, Estrada do Conde 6000, Cx. Postal 47, Eldorado do Sul, RS 92990-000, Brazil. E-mail: proehe@ufrgs.br Pseudorabies or Aujeszky’s disease (AD), caused by pseudorabies virus (PRV) is a major concern in swine production. In the state of Rio Grande do Sul, Brazil, AD was only detected in 1954, in cattle. In 2003 two outbreaks of encephalitis occurred on the northern region of the state, close to the border with the state of Santa Catarina. Pseudorabies virus (PRV) was isolated from distinct farms within the region and subjected to antigenic and genomic analyses. These isolates were compared with prototype strains NIA-3 and NP. Antigenic characterization with a panel of monoclonal antibodies (Mabs) directed to viral glycoproteins (gB, gC, gD and gE,) was performed by an imunoperoxidase monolayer assay (IPMA) on infected cell monolayers. Genomic characterization was carried out by restriction enzyme analysis (REA) of the whole DNA viral genome with Bam HI. The antigenic profile of the eight isolates from Rio Grande do Sul as well as strains NIA-3 and NP were similar. REA analysis revealed that all isolates from Rio Grande do Sul displayed a genomic type II arrangement, a genotype often found in other outbreaks of AD previously reported in other Brazilian states. The results obtained suggest that the eight isolates examined here were similar.

Abstract in English:

Groff F.H.S., Merlo M.A., Stoll P.A., Stepan A.L., Weiblen R. & Flores E.F. 2005. [Epidemiology and control of pseudorabies outbreaks in the state of Rio Grande do Sul, Brazil, 2003.] Epidemiologia e controle dos focos da doença de Aujeszky no Rio Grande do Sul, em 2003. Pesquisa Veterinária Brasileira 25(1):25-30. Depto Medicina Veterinária Preventiva, Universi-dade Federal de Santa Maria (UFSM), Santa Maria, RS 97105-900, Brazil. E-mail: flores@ccr.ufsm.br Aujeszky’s disease (AD) or pseudorabies is an important viral disease of swine and has significant economic impact on the pig industry worldwide. The infection produces direct and indirect economic losses, mainly due to restrictions to international trade of swine products. Since the beginning of the 20th century, AD had been notified in several Brazilian regions, yet the state of Rio Grande do Sul (RS) remained as a “provisionally free” area under the International Organization of Epizooties (OIE) guidelines. In 2003, two outbreaks were notified in swine herds located in northern RS, boundary with Santa Catarina, a state where the infection is endemic. Control/eradication measures consisted in tracing back all swine movements, quarantine and eradication of the affected herds by sanitary slaughter under official inspection. In outbreak # 1 (Pinheirinho do Vale, January/2003) six herds were affected, one of which presented animals with clinical signs. Starting from this outbreak, 146 herds and 42.399 pigs were traced back, six herds (7.822 animals) being depopulated – the herd where the index outbreak plus five other herds with positive serology. The outbreak # 2 (Aratiba, September/2003) resulted in a wider spread of the infection, involving another three counties and 77 herds (9 with clinical signs, 68 with positive serology). From the index case, 109.316 pigs in 630 herds were traced back; 28.443 animals from positive herds were slaughtered. Total numbers reached 151.715 traced back animals in 776 herds; 71 herds were found serologically positive. The adopted measures were efficient to eliminate the outbreaks and to stop the disease spread to neighboring areas, reestablishing the sanitary status previous to the outbreaks.

• Aujeszky’s disease pseudorabies ADV PRV epidemiology.

Abstract in Portuguese:

Groff F.H.S., Merlo M.A., Stoll P.A., Stepan A.L., Weiblen R. & Flores E.F. 2005. [Epidemiology and control of pseudorabies outbreaks in the state of Rio Grande do Sul, Brazil, 2003.] Epidemiologia e controle dos focos da doença de Aujeszky no Rio Grande do Sul, em 2003. Pesquisa Veterinária Brasileira 25(1):25-30. Depto Medicina Veterinária Preventiva, Universi-dade Federal de Santa Maria (UFSM), Santa Maria, RS 97105-900, Brazil. E-mail: flores@ccr.ufsm.br Aujeszky’s disease (AD) or pseudorabies is an important viral disease of swine and has significant economic impact on the pig industry worldwide. The infection produces direct and indirect economic losses, mainly due to restrictions to international trade of swine products. Since the beginning of the 20th century, AD had been notified in several Brazilian regions, yet the state of Rio Grande do Sul (RS) remained as a “provisionally free” area under the International Organization of Epizooties (OIE) guidelines. In 2003, two outbreaks were notified in swine herds located in northern RS, boundary with Santa Catarina, a state where the infection is endemic. Control/eradication measures consisted in tracing back all swine movements, quarantine and eradication of the affected herds by sanitary slaughter under official inspection. In outbreak # 1 (Pinheirinho do Vale, January/2003) six herds were affected, one of which presented animals with clinical signs. Starting from this outbreak, 146 herds and 42.399 pigs were traced back, six herds (7.822 animals) being depopulated – the herd where the index outbreak plus five other herds with positive serology. The outbreak # 2 (Aratiba, September/2003) resulted in a wider spread of the infection, involving another three counties and 77 herds (9 with clinical signs, 68 with positive serology). From the index case, 109.316 pigs in 630 herds were traced back; 28.443 animals from positive herds were slaughtered. Total numbers reached 151.715 traced back animals in 776 herds; 71 herds were found serologically positive. The adopted measures were efficient to eliminate the outbreaks and to stop the disease spread to neighboring areas, reestablishing the sanitary status previous to the outbreaks.