PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

Diel D.G., Fonseca E.T., Souza S.F., Mazzanti A., Bauermann F., Weiblen R. & Flores E.F. 2005. [Bovine herpesvirus 5 may use the olfactory and trigeminal pathways to invade the central nervous system of rabbits, depending upon the route of inoculation.] O Herpesvírus bovino tipo 5 (BoHV-5) pode utilizar as rotas olfatória ou trigeminal para invadir o sistema nervoso central de coelhos, dependendo da via de inoculação. Pesquisa Veterinária Brasileira 25(3):164-170. Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. E-mail: flores@ccr.ufsm.br Bovine herpesvirus type 5 (BoHV-5) is a major etiological agent of meningoencephalitis in cattle. Following replication in the nasal mucosa, viral invasion of the brain is thought to occur mainly by the olfactory pathway. To address the role of this pathway in the pathogenesis of neurological infection in a laboratory model, 30 days old rabbits had the main olfactory bulbs (MOBs) surgically removed and were subsequently inoculated intranasally (IN) or conjunctivally (IC) with a highly neurovirulent BoHV-5 strain (SV-507). Following IN inoculation, 10 out of 10 (100 %) control rabbits developed neurological disease. The clinical onset ranged from day 5 to 10 post-inoculation (pi, average 7.5 days); nine being euthanized in extremis and one recovering after a mild clinical course. In contrast, only one rabbit (9.1 %) of the group lacking the MOBs (n=11) developed neurological disease (onset at day 17 pi). Dexamethasone administration to the survivors (n=10) at day 50pi was followed by virus shedding in nasal and/or ocular secretions by 8 animals, demonstrating that the virus was able to reach the trigeminal ganglia (TG) during acute infection. These results demonstrate that the olfactory route provides the main, yet not the sole access to the brain of rabbits following IN inoculation. To address the role of a second pathway, groups of control (n=12) or MOB-lacking rabbits (n=12) were inoculated into the conjunctival sac (IC), following which the virus would be expected to use the ophtalmic branch of the trigeminal nerve to reach the brain. Ten control rabbits (83.3 %) developed neurological disease upon IC inoculation (onset 15.3 days [11 to 20]). Previous ablation of the MOBs did not affect the frequency and course of neurological disease: ten out of 12 rabbits (83.3 %) lacking the MOBs developed neurological disease (onset 9 to 15 dpi, average: 12.7 days) upon IC inoculation. These results demonstrate that both IN and IC routes may operate in the transport of BoHV-5 to the brain of experimentally infected rabbits, depending on the route of inoculation. IN inoculation results in a fast and efficient transport by the olfactory pathway, the trigeminal route providing an alternative, much slower and less efficient transport; IC inoculation results in efficient viral transport by the trigeminal route, yet with a delayed kinetics comparing to the transport provided by the olfactory pathway.

• Bovine herpesvirus type 5 BoHV-5 neurological infection rabbits.

Abstract in Portuguese:

Diel D.G., Fonseca E.T., Souza S.F., Mazzanti A., Bauermann F., Weiblen R. & Flores E.F. 2005. [Bovine herpesvirus 5 may use the olfactory and trigeminal pathways to invade the central nervous system of rabbits, depending upon the route of inoculation.] O Herpesvírus bovino tipo 5 (BoHV-5) pode utilizar as rotas olfatória ou trigeminal para invadir o sistema nervoso central de coelhos, dependendo da via de inoculação. Pesquisa Veterinária Brasileira 25(3):164-170. Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. E-mail: flores@ccr.ufsm.br Bovine herpesvirus type 5 (BoHV-5) is a major etiological agent of meningoencephalitis in cattle. Following replication in the nasal mucosa, viral invasion of the brain is thought to occur mainly by the olfactory pathway. To address the role of this pathway in the pathogenesis of neurological infection in a laboratory model, 30 days old rabbits had the main olfactory bulbs (MOBs) surgically removed and were subsequently inoculated intranasally (IN) or conjunctivally (IC) with a highly neurovirulent BoHV-5 strain (SV-507). Following IN inoculation, 10 out of 10 (100 %) control rabbits developed neurological disease. The clinical onset ranged from day 5 to 10 post-inoculation (pi, average 7.5 days); nine being euthanized in extremis and one recovering after a mild clinical course. In contrast, only one rabbit (9.1 %) of the group lacking the MOBs (n=11) developed neurological disease (onset at day 17 pi). Dexamethasone administration to the survivors (n=10) at day 50pi was followed by virus shedding in nasal and/or ocular secretions by 8 animals, demonstrating that the virus was able to reach the trigeminal ganglia (TG) during acute infection. These results demonstrate that the olfactory route provides the main, yet not the sole access to the brain of rabbits following IN inoculation. To address the role of a second pathway, groups of control (n=12) or MOB-lacking rabbits (n=12) were inoculated into the conjunctival sac (IC), following which the virus would be expected to use the ophtalmic branch of the trigeminal nerve to reach the brain. Ten control rabbits (83.3 %) developed neurological disease upon IC inoculation (onset 15.3 days [11 to 20]). Previous ablation of the MOBs did not affect the frequency and course of neurological disease: ten out of 12 rabbits (83.3 %) lacking the MOBs developed neurological disease (onset 9 to 15 dpi, average: 12.7 days) upon IC inoculation. These results demonstrate that both IN and IC routes may operate in the transport of BoHV-5 to the brain of experimentally infected rabbits, depending on the route of inoculation. IN inoculation results in a fast and efficient transport by the olfactory pathway, the trigeminal route providing an alternative, much slower and less efficient transport; IC inoculation results in efficient viral transport by the trigeminal route, yet with a delayed kinetics comparing to the transport provided by the olfactory pathway.

Abstract in English:

Flores E.F., Weiblen R., Vogel F.S.F., Roehe P.M., Alfieri A.A. & Pituco E.M. 2005. [Bovine viral diarrhea virus (BVDV) infection in Brazil: history, current situation and perspectives.] A infecção pelo vírus da Diarréia Viral Bovina (BVDV) no Brasil - histórico, situação atual e perspectivas. Pesquisa Veterinária Brasileira 25(3):125-134. Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. E-mail: flores@ccr.ufsm.br Bovine viral diarrhea virus (BVDV) is one of the most important pathogens of cattle worldwide. BVDV infection and associated diseases have been reported in Brazil since the late 1960ties. Several serological, virological, clinical and pathological reports demonstrate the widespread distribution of BVDV infection among Brazilian cattle. In addition to variable levels of positive serology in beef and dairy cattle, BVDV antibodies have been occasionally detected in swine, wild boars, goats, cervids and water buffaloes. BVDV infection has been diagnosed in aborted fetuses, buffy coats of persistently infected (PI) animals, clinical specimens from animals suffering from different clinical syndromes, semen of bulls of artificial insemination (AI) centers, in healthy fetuses and in commercial fetal bovine serum and/or cultured cells. About 50 isolates have been genetically and/or antigenically characterized up to date, whilst roughly an equivalent number of isolates awaits characterization. Most of the characterized isolates belong to BVDV-1 genotype, non-cytopathic (NCP) biotype, yet some BVDV-2 (and some CP BVDV) have been identified as well. Brazilian BVDV isolates display a high antigenic variability and are markedly different from North American vaccine strains. A few inactivated, polyvalent vaccines are currently licensed in the country, yet vaccination is still incipient in many regions: only about 2.5 million doses were marketed in 2003. The low serological cross-reactivity between vaccine strains and field isolates has recently stimulated national industries to develop vaccines containing Brazilian BVDV-1 and BVDV-2 strains. The overall knowledge about BVDV infection in Brazil has grown considerably in the last years, due to an increasing number of laboratories performing diagnosis and research. Studies on the pathogenesis, serological and molecular epidemiology and production of reagents for diagnosis have contributed decisively for the recent growing knowledge on BVDV infections in the country.

• Bovine viral diarrhea virus BVDV epidemiology diagnostic.

Abstract in Portuguese:

Flores E.F., Weiblen R., Vogel F.S.F., Roehe P.M., Alfieri A.A. & Pituco E.M. 2005. [Bovine viral diarrhea virus (BVDV) infection in Brazil: history, current situation and perspectives.] A infecção pelo vírus da Diarréia Viral Bovina (BVDV) no Brasil - histórico, situação atual e perspectivas. Pesquisa Veterinária Brasileira 25(3):125-134. Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. E-mail: flores@ccr.ufsm.br Bovine viral diarrhea virus (BVDV) is one of the most important pathogens of cattle worldwide. BVDV infection and associated diseases have been reported in Brazil since the late 1960ties. Several serological, virological, clinical and pathological reports demonstrate the widespread distribution of BVDV infection among Brazilian cattle. In addition to variable levels of positive serology in beef and dairy cattle, BVDV antibodies have been occasionally detected in swine, wild boars, goats, cervids and water buffaloes. BVDV infection has been diagnosed in aborted fetuses, buffy coats of persistently infected (PI) animals, clinical specimens from animals suffering from different clinical syndromes, semen of bulls of artificial insemination (AI) centers, in healthy fetuses and in commercial fetal bovine serum and/or cultured cells. About 50 isolates have been genetically and/or antigenically characterized up to date, whilst roughly an equivalent number of isolates awaits characterization. Most of the characterized isolates belong to BVDV-1 genotype, non-cytopathic (NCP) biotype, yet some BVDV-2 (and some CP BVDV) have been identified as well. Brazilian BVDV isolates display a high antigenic variability and are markedly different from North American vaccine strains. A few inactivated, polyvalent vaccines are currently licensed in the country, yet vaccination is still incipient in many regions: only about 2.5 million doses were marketed in 2003. The low serological cross-reactivity between vaccine strains and field isolates has recently stimulated national industries to develop vaccines containing Brazilian BVDV-1 and BVDV-2 strains. The overall knowledge about BVDV infection in Brazil has grown considerably in the last years, due to an increasing number of laboratories performing diagnosis and research. Studies on the pathogenesis, serological and molecular epidemiology and production of reagents for diagnosis have contributed decisively for the recent growing knowledge on BVDV infections in the country.

Abstract in English:

Rech R.R., Schild A.L., Driemeier D., Garmatz S.L., Oliveira F.N., Riet-Correa F. & Barros C.S.L. 2005. [Malignant catarrhal fever in cattle in Rio Grande do Sul, Brazil: Epidemiology, clinical signs and pathology.] Febre catarral maligna em bovinos no Rio Grande do Sul: epidemiologia, sinais clínicos e patologia. Pesquisa Veterinária Brasileira 25(2):97-105. Depto Patologia, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. E-mail: claudioslbarros@uol.com.br The epidemiology, clinical signs, necropsy and histopathological findings in cases of malignant catarrhal fever (MCF) occurring in cattle from 15 farms in Rio Grande do Sul, Brazil, from 1973 to 2003, are described. In 9 instances (60%) the disease occurred as sporadic cases affecting 1-3 cattle whereas in six farms (40%) MCF occurred as epizootics involving several cattle in each affected herd. Morbidity rates ranged from 2.4% to 20% and lethality rates were 83.3% and 100%. Cattle of all ages and both sexes were affected. Where the information was available (9 farms) sheep were in contact with affected cattle and cases of MCF occurred more frequently in spring and summer. Clinical courses were acute or subacute and clinical signs included fever, nasal and ocular discharges, conjunctivitis, drooling, hematuria, necrosis and blunting of buccal papillae, enlargement of lymph nodes, diarrhea and neurological disturbances. Necropsy findings included opaque corneas, reddening, erosions and ulcerations in several mucous membranes of the alimentary, respiratory and urogenital tracts, and the conjunctiva; enlargement and haemorrhage of lymph nodes and multiple white foci in the renal cortices and in the hepatic portal triads. Crustous dermatitis was observed in some cases. Main histopathological findings included vasculitis, necrosis of the surface epithelia and accumulation of inflammatory cells in several organs. Vasculitis were associated with fibrinoid necrosis of the medial layer of arteries and inflammatory cells included lymphoblasts, lymphocytes, plasma cells and macrophages.

• Infectious diseases epidemiology viral diseases diseases of cattle pathology.

Abstract in Portuguese:

Rech R.R., Schild A.L., Driemeier D., Garmatz S.L., Oliveira F.N., Riet-Correa F. & Barros C.S.L. 2005. [Malignant catarrhal fever in cattle in Rio Grande do Sul, Brazil: Epidemiology, clinical signs and pathology.] Febre catarral maligna em bovinos no Rio Grande do Sul: epidemiologia, sinais clínicos e patologia. Pesquisa Veterinária Brasileira 25(2):97-105. Depto Patologia, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. E-mail: claudioslbarros@uol.com.br The epidemiology, clinical signs, necropsy and histopathological findings in cases of malignant catarrhal fever (MCF) occurring in cattle from 15 farms in Rio Grande do Sul, Brazil, from 1973 to 2003, are described. In 9 instances (60%) the disease occurred as sporadic cases affecting 1-3 cattle whereas in six farms (40%) MCF occurred as epizootics involving several cattle in each affected herd. Morbidity rates ranged from 2.4% to 20% and lethality rates were 83.3% and 100%. Cattle of all ages and both sexes were affected. Where the information was available (9 farms) sheep were in contact with affected cattle and cases of MCF occurred more frequently in spring and summer. Clinical courses were acute or subacute and clinical signs included fever, nasal and ocular discharges, conjunctivitis, drooling, hematuria, necrosis and blunting of buccal papillae, enlargement of lymph nodes, diarrhea and neurological disturbances. Necropsy findings included opaque corneas, reddening, erosions and ulcerations in several mucous membranes of the alimentary, respiratory and urogenital tracts, and the conjunctiva; enlargement and haemorrhage of lymph nodes and multiple white foci in the renal cortices and in the hepatic portal triads. Crustous dermatitis was observed in some cases. Main histopathological findings included vasculitis, necrosis of the surface epithelia and accumulation of inflammatory cells in several organs. Vasculitis were associated with fibrinoid necrosis of the medial layer of arteries and inflammatory cells included lymphoblasts, lymphocytes, plasma cells and macrophages.

Abstract in English:

Simionatto S., Lima-Rosa C.A.V., Rubin L.L. & Canal C.W. 2005. [A nested-PCR protocol for detection of the chicken anemia virus.] Um protocolo de “nested-PCR” para detecção do virus da anemia das galinhas. Pesquisa Veterinária Brasileira 25(2):106-110. Laboratório de Virologia, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: claudio.canal@ufrgs.br This paper reports a nested polymerase chain reaction (nested-PCR) protocol for detection of chicken anemia virus (CAV), the causal agent of infectious chicken anemia. For DNA extraction from clinical samples, a method based on guanidine thiocyanate was found more sensitive and practical than other extraction protocols tested. The pair of primers used in the initial PCR targeted a 664 bp fragment on the VP1 gene. The primers for the internal PCR targeted a fragment of 520 bp. The specificity of the primers was evaluated on samples of CAV controlled flocks. Thirty different viruses and bacteria isolated from chickens did not give rise to any amplification product in the assay. The sensitivity of the nested-PCR was determined on serial dilutions of a CAV vaccine. The nested-PCR was more sensitive than a one step PCR and was able to detect at least 0.16 TCID50 of the vaccine strain. In addition, the protocol employed here detected viral DNA from tissues, sera and litter from flocks with or without clinical signs of disease. It is concluded that the nested-PCR protocol described here is more sensitive, faster and less cumbersome than virus isolation in cell culture as a diagnostic technique for detection of CAV.

• Chicken anemia virus CAV molecular diagnosis nested-PCR.

Abstract in Portuguese:

Simionatto S., Lima-Rosa C.A.V., Rubin L.L. & Canal C.W. 2005. [A nested-PCR protocol for detection of the chicken anemia virus.] Um protocolo de “nested-PCR” para detecção do virus da anemia das galinhas. Pesquisa Veterinária Brasileira 25(2):106-110. Laboratório de Virologia, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: claudio.canal@ufrgs.br This paper reports a nested polymerase chain reaction (nested-PCR) protocol for detection of chicken anemia virus (CAV), the causal agent of infectious chicken anemia. For DNA extraction from clinical samples, a method based on guanidine thiocyanate was found more sensitive and practical than other extraction protocols tested. The pair of primers used in the initial PCR targeted a 664 bp fragment on the VP1 gene. The primers for the internal PCR targeted a fragment of 520 bp. The specificity of the primers was evaluated on samples of CAV controlled flocks. Thirty different viruses and bacteria isolated from chickens did not give rise to any amplification product in the assay. The sensitivity of the nested-PCR was determined on serial dilutions of a CAV vaccine. The nested-PCR was more sensitive than a one step PCR and was able to detect at least 0.16 TCID50 of the vaccine strain. In addition, the protocol employed here detected viral DNA from tissues, sera and litter from flocks with or without clinical signs of disease. It is concluded that the nested-PCR protocol described here is more sensitive, faster and less cumbersome than virus isolation in cell culture as a diagnostic technique for detection of CAV.

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:

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.

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.