PESQUISA VETERINÁRIA BRASILEIRA

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.

Abstract in English:

Elias F., Schild A.L. & Riet-Correa F. 2004. [Bovine herpesvirus type-5 meningoencephalitis and malacia: histological lesions distribution in the central nervous system of naturally infected cattle.] Meningoencefalite e encefalomalacia por Herpesvírus bovino-5: distribuição das lesões no sistema nervoso central de bovinos naturalmente infectados. Pesquisa Veterinária Brasileira 24(3):123-131. Laboratório Regional de Diagnóstico, Faculdade de Veterinária, UFPel, Cx. Postal 354, Pelotas, RS 96010-900, Brazil. E-mail: alschild@terra.com.br The distribution of the histological lesions in the central nervous system (CNS) of cattle naturally infected by bovine herpes virus type-5 (BHV-5) was determined in 12 affected calves from 10 outbreaks of the disease diagnosed by the Regional Diagnostic Laboratory (LRD) at Pelotas University, from 1986 to 2003. The epidemiological data, clinical signs and duration of clinical course were obtained from the files of LRD. Transversal sections were performed at different levels in 10% formalin-fixed CNS. The sections were made in the frontal, parietal, temporal e occipital lobes of the telencephalic hemispheres, basal ganglia and internal capsule, thalamus, anterior colliculus, pons, cerebellar peduncles, cerebellum, medulla oblongata and cervical spinal cord. Paraffin embedded tissues were sectioned and stained with hematoxylin and eosin. The severity and distribution of the inflammatory and malacic lesions were evaluated in all sections. These lesions were related with the epidemiological and clinical aspects of the disease. The outbreaks of the disease were observed in different seasons of the year. Affected animals were 2 to 24-month-old, of different breeds and both sexes. Gross lesions characterized by yellow and depressed areas in the cerebral cortex were observed in five calves. In two of them, similar lesions were additionally observed in thalamus, basal nuclei, and internal capsule. Congestion and multifocal hemorrhages were observed in most cases. The histological lesions were characterized by non-suppurative meningoencephalitis in all sections of CNS, but more severe in the frontal cortex. Focal or widespread malacia with infiltration of Gitter cells were observed in all sections of cerebral cortex, basal ganglia, internal capsule, and thalamus. In some cases mild malacia was also observed in the rostral colliculi, pons, medulla, cerebellum and cervical spinal cord. Intranuclear inclusion bodies were seen in all cases studied; they were frequent in regions of the cerebral cortex near mild to moderate inflammatory or malacic lesions. In two cases the inclusion bodies were also seen in the basal ganglia and thalamus. The severity of the histological lesions was not proportional with the clinical course of the disease. The presence of lesions of malacia in different regions of the CNS, an aspect not mentioned in most reports of BHV-5 infections, could be due to variable pathogenicity of different virus isolates. Alternatively, it is possible that BHV-5 encephalitis occurs due to the reactivation of the virus in cattle previously affected by polioencefalomacia; this last sequence of events was already demonstrated experimentally by our research group.

• Bovine herpesvirus type-5 BHV-5 cattle meningoencephalitis polioencephalomalacia.

Abstract in Portuguese:

Elias F., Schild A.L. & Riet-Correa F. 2004. [Bovine herpesvirus type-5 meningoencephalitis and malacia: histological lesions distribution in the central nervous system of naturally infected cattle.] Meningoencefalite e encefalomalacia por Herpesvírus bovino-5: distribuição das lesões no sistema nervoso central de bovinos naturalmente infectados. Pesquisa Veterinária Brasileira 24(3):123-131. Laboratório Regional de Diagnóstico, Faculdade de Veterinária, UFPel, Cx. Postal 354, Pelotas, RS 96010-900, Brazil. E-mail: alschild@terra.com.br The distribution of the histological lesions in the central nervous system (CNS) of cattle naturally infected by bovine herpes virus type-5 (BHV-5) was determined in 12 affected calves from 10 outbreaks of the disease diagnosed by the Regional Diagnostic Laboratory (LRD) at Pelotas University, from 1986 to 2003. The epidemiological data, clinical signs and duration of clinical course were obtained from the files of LRD. Transversal sections were performed at different levels in 10% formalin-fixed CNS. The sections were made in the frontal, parietal, temporal e occipital lobes of the telencephalic hemispheres, basal ganglia and internal capsule, thalamus, anterior colliculus, pons, cerebellar peduncles, cerebellum, medulla oblongata and cervical spinal cord. Paraffin embedded tissues were sectioned and stained with hematoxylin and eosin. The severity and distribution of the inflammatory and malacic lesions were evaluated in all sections. These lesions were related with the epidemiological and clinical aspects of the disease. The outbreaks of the disease were observed in different seasons of the year. Affected animals were 2 to 24-month-old, of different breeds and both sexes. Gross lesions characterized by yellow and depressed areas in the cerebral cortex were observed in five calves. In two of them, similar lesions were additionally observed in thalamus, basal nuclei, and internal capsule. Congestion and multifocal hemorrhages were observed in most cases. The histological lesions were characterized by non-suppurative meningoencephalitis in all sections of CNS, but more severe in the frontal cortex. Focal or widespread malacia with infiltration of Gitter cells were observed in all sections of cerebral cortex, basal ganglia, internal capsule, and thalamus. In some cases mild malacia was also observed in the rostral colliculi, pons, medulla, cerebellum and cervical spinal cord. Intranuclear inclusion bodies were seen in all cases studied; they were frequent in regions of the cerebral cortex near mild to moderate inflammatory or malacic lesions. In two cases the inclusion bodies were also seen in the basal ganglia and thalamus. The severity of the histological lesions was not proportional with the clinical course of the disease. The presence of lesions of malacia in different regions of the CNS, an aspect not mentioned in most reports of BHV-5 infections, could be due to variable pathogenicity of different virus isolates. Alternatively, it is possible that BHV-5 encephalitis occurs due to the reactivation of the virus in cattle previously affected by polioencefalomacia; this last sequence of events was already demonstrated experimentally by our research group.

Abstract in English:

Lemos E.R.S., D'Andrea P.S., Bonvicino C. R., Famadas K. M., Padula P., Cavalcanti A.A. & Schatzmayr H.G. 2004. Evidence of hantavirus infection in wild rodents captured in a rural area of the state of São Paulo, Brazil. Pesquisa Veterinária Brasileira 24(2):71-73. Depto Virologia, Instituto Oswaldo Cruz, Av. Brasil 4365, Pavilhão Rocha Lima, 5º andar, Manguinhos, Rio de Janeiro, RJ, 21045-900, Brazil. E-mail: elemos@ioc.fiocruz.br Hantaviruses are the etiological agents of the Hantavirus Cardio-Pulmonary Syndrome, a serious rodent-borne disease in Brazil. In order to investigate the occurrence of hantavirus infection in wild rodents, a survey was conducted in three different suburban areas of the municipality of Pedreira, State of São Paulo, Brazil. Of the 145 wild animals captured belonging to 12 different species identified by morphology and karyological analysis, 107 were rodents of the following species: Akodon montensis, Bolomys lasiurus, Calomys tener, Oligoryzomys nigripes, Oligoryzomys flavescens, and Myocastor coypus. Blood samples from these rodents were assayed for the presence of antibodies against hantavirus by IgG ELISA using Andes recombinant nucleocapsid antigen. Antibody reactive to Andes virus was found in two different species, O. nigripes and O. flavescens. These results indicate a potential risk for hantavirus transmission to humans in this area, where reservoir rodents are present in peridomestic settings.

• Hantavirus rodent survey Pedreira São Paulo Brazil.

Abstract in Portuguese:

Lemos E.R.S., D'Andrea P.S., Bonvicino C. R., Famadas K. M., Padula P., Cavalcanti A.A. & Schatzmayr H.G. 2004. Evidence of hantavirus infection in wild rodents captured in a rural area of the state of São Paulo, Brazil. Pesquisa Veterinária Brasileira 24(2):71-73. Depto Virologia, Instituto Oswaldo Cruz, Av. Brasil 4365, Pavilhão Rocha Lima, 5º andar, Manguinhos, Rio de Janeiro, RJ, 21045-900, Brazil. E-mail: elemos@ioc.fiocruz.br Hantaviruses are the etiological agents of the Hantavirus Cardio-Pulmonary Syndrome, a serious rodent-borne disease in Brazil. In order to investigate the occurrence of hantavirus infection in wild rodents, a survey was conducted in three different suburban areas of the municipality of Pedreira, State of São Paulo, Brazil. Of the 145 wild animals captured belonging to 12 different species identified by morphology and karyological analysis, 107 were rodents of the following species: Akodon montensis, Bolomys lasiurus, Calomys tener, Oligoryzomys nigripes, Oligoryzomys flavescens, and Myocastor coypus. Blood samples from these rodents were assayed for the presence of antibodies against hantavirus by IgG ELISA using Andes recombinant nucleocapsid antigen. Antibody reactive to Andes virus was found in two different species, O. nigripes and O. flavescens. These results indicate a potential risk for hantavirus transmission to humans in this area, where reservoir rodents are present in peridomestic settings.

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:

Lima M., Flores E.F., Weiblen R., Flores F.S.F. & Arenhart S. 2004. [Characterization of bovine viral diarrhea virus (BVDV) types 1 and 2 isolates for use in vaccines.] Caracterização de amostras atenuadas do vírus da Diarréia Viral Bovina (BVDV) tipos 1 e 2 para uso em vacinas. Pesquisa Veterinária Brasileira 24(1):35-42. Depto Medicina Veterinária Preventiva, Centro de Ciências Rurais, Universidade Federal de Santa Maria, Santa Maria, RS 97105-900, Brazil. E-mail: flores@ccr.ufsm.br This article reports the characterization of two cytopathic isolates of bovine viral diarrhea virus (BVDV-1: IBSP-2; BVDV-2:SV-253) submitted to multiple passages (n=30) in tissue culture associated with ultraviolet irradiation. The vaccine candidate strains were characterized in vitro (plaque size and morphology, growth kinetics and antigenic profile) and in vivo (attenuation and serological response in calves). In vitro characterization of biologically cloned viruses obtained at passages 0, 1, 10, 20 and 30 demonstrated that the attenuation process did not significantly affect the phenotypic and antigenic properties of the viruses. No major differences in plaque size and morphology and in the growth kinetics in tissue culture were observed among the viruses obtained at different passages. Likewise, the antigenic profile of these viruses did not change upon successive passages in tissue culture, as ascertained by the pattern of binding by 48 monoclonal antibodies (mAbs). Intramuscular inoculation of both viruses (IBSP-2: 107.3 TCID50; SV-253: 106.8 TCID50) at passage 30 (p30) in twelve 15 months old heifers did not produce clinical signs, demonstrating the attenuation of the viruses. Following inoculation, infectious virus was detected in leucocytes of most inoculated animals (10/12) between days 3 and 6 post-inoculation (pi) and in nasal secretions of three animals (days 4, 7 and 8pi). However, the vaccine viruses were not transmitted to three seronegative calves maintained as sentinels. All vaccinated calves seroconverted at day 14 post-vaccination. A moderate to high serum neutralizing response against five Brazilian BVDV-1 (titers from 80 to ³1,280) and four Brazilian BVDV-2 isolates (titers from 20 to 640) was observed at day 33 post-vaccination (pv). In general, the highest titers were observed against the Brazilian BVDV-1 isolates. At day 240 post-vaccination, the animals received a booster administration (IBSP-2: 107.3 TCID50 and SV-253: 106.8 TCID50). Revaccination resulted in a strong anamnestic response in most animals, with increasing antibody titers mainly to BVDV-2. These are promising results towards the future use of these strains in modified-live vaccines for the control of BVDV infection in Brazil.

• Bovine viral diarrhea virus (BVDV) modified-live vaccine attenuation.

Abstract in Portuguese:

Lima M., Flores E.F., Weiblen R., Flores F.S.F. & Arenhart S. 2004. [Characterization of bovine viral diarrhea virus (BVDV) types 1 and 2 isolates for use in vaccines.] Caracterização de amostras atenuadas do vírus da Diarréia Viral Bovina (BVDV) tipos 1 e 2 para uso em vacinas. Pesquisa Veterinária Brasileira 24(1):35-42. Depto Medicina Veterinária Preventiva, Centro de Ciências Rurais, Universidade Federal de Santa Maria, Santa Maria, RS 97105-900, Brazil. E-mail: flores@ccr.ufsm.br This article reports the characterization of two cytopathic isolates of bovine viral diarrhea virus (BVDV-1: IBSP-2; BVDV-2:SV-253) submitted to multiple passages (n=30) in tissue culture associated with ultraviolet irradiation. The vaccine candidate strains were characterized in vitro (plaque size and morphology, growth kinetics and antigenic profile) and in vivo (attenuation and serological response in calves). In vitro characterization of biologically cloned viruses obtained at passages 0, 1, 10, 20 and 30 demonstrated that the attenuation process did not significantly affect the phenotypic and antigenic properties of the viruses. No major differences in plaque size and morphology and in the growth kinetics in tissue culture were observed among the viruses obtained at different passages. Likewise, the antigenic profile of these viruses did not change upon successive passages in tissue culture, as ascertained by the pattern of binding by 48 monoclonal antibodies (mAbs). Intramuscular inoculation of both viruses (IBSP-2: 107.3 TCID50; SV-253: 106.8 TCID50) at passage 30 (p30) in twelve 15 months old heifers did not produce clinical signs, demonstrating the attenuation of the viruses. Following inoculation, infectious virus was detected in leucocytes of most inoculated animals (10/12) between days 3 and 6 post-inoculation (pi) and in nasal secretions of three animals (days 4, 7 and 8pi). However, the vaccine viruses were not transmitted to three seronegative calves maintained as sentinels. All vaccinated calves seroconverted at day 14 post-vaccination. A moderate to high serum neutralizing response against five Brazilian BVDV-1 (titers from 80 to ³1,280) and four Brazilian BVDV-2 isolates (titers from 20 to 640) was observed at day 33 post-vaccination (pv). In general, the highest titers were observed against the Brazilian BVDV-1 isolates. At day 240 post-vaccination, the animals received a booster administration (IBSP-2: 107.3 TCID50 and SV-253: 106.8 TCID50). Revaccination resulted in a strong anamnestic response in most animals, with increasing antibody titers mainly to BVDV-2. These are promising results towards the future use of these strains in modified-live vaccines for the control of BVDV infection in Brazil.

Abstract in English:

Spilki F.R, Esteves P.A., Lima M., Franco A.C., Chiminazzo C., Flores E.F., Weiblen R., Driemeier D. & Roehe P.M. 2004. Comparative pathogenicity of bovine herpesviruses type 1 (BHV-1) subtypes 1 (BHV-1.1) and 2a (BHV-1.2a). Pesquisa Veterinária Brasileira 24(1):43-49. Centro de Pesquisas Desidério Finamor, Fepagro Saúde Animal, Cx. Postal 47, Eldorado do Sul, RS 92990-000, Brazil. E-mail: proehe@ufrgs.br The study aimed to examine the capacity of two bovine herpesvirus type 1 (BHV-1) isolates of different subtypes (EVI 123/96, BHV-1.1; SV265/98, BHV-1.2a) to induce respiratory disease in calves. These two isolates are representative of the BHV-1 subtypes prevalent in Brazil. Viral subtypes were confirmed by monoclonal antibody analysis and by restriction enzyme digestion of viral genomes. The viruses were inoculated intranasally into seven 3 months old calves (four with BHV-1.1, three with BHV-1.2a). Three other calves of identical age and condition were kept as uninfected controls. In both groups of infected calves, the clinical signs observed were consistent with typical infectious bovine rhinothracheitis (IBR), including pyrexia, apathy, anorexia, nasal and ocular mucopurulent discharges, erosions on the nasal mucosa, conjunctivitis, lachrymation, redness of nasal mucosa, dyspnoea, coughing, tracheal stridor and enlargement of retropharingeal, submandibular and cervical lymphnodes. No significant differences were observed between the clinical scores attributed to both groups. Virus shedding in nasal and ocular secretions were also similar, apart from a significant difference in nasal virus shedding on day 1 to 3 post-inoculation, which was higher for BHV-1.1 than for BHV-1.2a. Following corticosteroid induced reactivation of the latent infection, recrudescence of clinical signs was also observed, with no significant differences on both groups. It was concluded that both subtypes BHV-1.1 and BHV-1.2a were able to induce clinically undistinguishable respiratory disease in calves, either subsequent to a primary infection or following reactivation.

• Infectious bovine rhinotracheitis IBR BHV-1.1 BHV-1.2a subtypes pathogenicity.

Abstract in Portuguese:

Spilki F.R, Esteves P.A., Lima M., Franco A.C., Chiminazzo C., Flores E.F., Weiblen R., Driemeier D. & Roehe P.M. 2004. Comparative pathogenicity of bovine herpesviruses type 1 (BHV-1) subtypes 1 (BHV-1.1) and 2a (BHV-1.2a). Pesquisa Veterinária Brasileira 24(1):43-49. Centro de Pesquisas Desidério Finamor, Fepagro Saúde Animal, Cx. Postal 47, Eldorado do Sul, RS 92990-000, Brazil. E-mail: proehe@ufrgs.br The study aimed to examine the capacity of two bovine herpesvirus type 1 (BHV-1) isolates of different subtypes (EVI 123/96, BHV-1.1; SV265/98, BHV-1.2a) to induce respiratory disease in calves. These two isolates are representative of the BHV-1 subtypes prevalent in Brazil. Viral subtypes were confirmed by monoclonal antibody analysis and by restriction enzyme digestion of viral genomes. The viruses were inoculated intranasally into seven 3 months old calves (four with BHV-1.1, three with BHV-1.2a). Three other calves of identical age and condition were kept as uninfected controls. In both groups of infected calves, the clinical signs observed were consistent with typical infectious bovine rhinothracheitis (IBR), including pyrexia, apathy, anorexia, nasal and ocular mucopurulent discharges, erosions on the nasal mucosa, conjunctivitis, lachrymation, redness of nasal mucosa, dyspnoea, coughing, tracheal stridor and enlargement of retropharingeal, submandibular and cervical lymphnodes. No significant differences were observed between the clinical scores attributed to both groups. Virus shedding in nasal and ocular secretions were also similar, apart from a significant difference in nasal virus shedding on day 1 to 3 post-inoculation, which was higher for BHV-1.1 than for BHV-1.2a. Following corticosteroid induced reactivation of the latent infection, recrudescence of clinical signs was also observed, with no significant differences on both groups. It was concluded that both subtypes BHV-1.1 and BHV-1.2a were able to induce clinically undistinguishable respiratory disease in calves, either subsequent to a primary infection or following reactivation.

Abstract in English:

ABSTRACT.- Spilki F.R., Franco A.C., Teixeira M.B., Esteves P.A., Schaefer R., Schmidt E., Lemos R.A. & Roehe P.M. 2003. Bovine herpesvirus type 5 (BHV-5) in a calf with rabies. Pesquisa Veterinária Brasileira 23(1):1-4. Centro de Pesquisas Veterinárias Desidério Finamor, FepagroSaúde Animal, Cx. Postal 2076, Porto Alegre, RS 90001-970, Brazil. The brain of an one year old mal e calf which died with signs of neurological disease was submitted to the laboratory for rabies diagnosis. Microscopical findings included moderate mielitis, mild meningoencephalitis with perivascular cell cuffing and Negri inclusion bodies in Purkinje cells of the cerebellum. Rabies virus infection was further confirmed by the direct fluorescent antibody test as well as by mouse inoculation. ln addition, a herpesvirus was isolated from brain tissues. The isolate was antigenic and genetically characterized as bovine herpesvirus type 5 (BHV-5). It was not possible to determine whether BHV-5 played an active role in the outcome of the infection, since, the virus might have been present in a latent form in neural tissues. This is the first report of a mixed rabies/ BHV-5 infection in calves.

• Bovine herpesvirus type 5 BHV-5 rabies Herpesvírus bovino tipo 5 raiva.

Abstract in Portuguese:

RESUMO.- Spilki F.R., Franco A.C., Teixeira M.B., Esteves P.A., Schaefer R., Schmidt E., Lemos R.A. & Roehe P.M. 2003. Bovine herpesvirus type 5 (BHV-5) in a calf with rabies. Pesquisa Veterinária Brasileira 23(1):1-4. [Herpesvírus bovino tipo 5 (BHV-5) em bovino infectado pelo vírus da raiva.] Centro de Pesquisas Veterinárias Desidério Finamor, FepagroSaúde Animal, Cx. Postal 2076, Porto Alegre, RS 90001-970, Brazil. O encéfalo de um bezerro macho, de um ano de idade, que morreu com sinais de doença neurológica, foi submetido ao laboratório para diagnóstico de raiva. O exame histopatológico revelou mielite moderada, meningoencefalite leve com infiltração perivascular e corpúsculos de Negri nas células de Purkinje do cerebelo. A infecção pelo vírus rábico foi ainda confirmada por imunofluorescência diretá e por inoculação em camundongos. Além disso, um herpesvírus foi isolado dos tecidos do encéfalo. O vírus isolado foi antigênica e geneticamente caracterizado como Herpesvírus bovino tipo 5 (BHV-5). Não foi possível determinar se o BHV-5 teve algum papel ativo no desfecho da enfermidade, uma vez que o vírus poderia estar presente em forma latente nos tecidos neurais. Este é o primeiro relato de uma infecção mista pelo vírus da raiva e BHV-5 em bovinos.