PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

Lima E.F., Riet-Correa F., Tabosa I.M., Dantas A.F.M., Medeiros J.M. & Sucupira Júnior G. 2005. [Polioencephalomalacia in goats and sheep in the semiarid region of northeastern Brazil.] Polioencefalomalacia em caprinos e ovinos na região semi-árida do Nordeste do Brasil. Pesquisa Veterinária Brasileira 25(1):9-14. Centro de Saúde e Tecnologia Rural, Universidade Federal de Campina Grande, Campus de Patos, 58700-000 Patos, PB, Brazil. E-mail: riet@cstr.ufcg.edu.br Seven outbreaks of polioencephalomalacia in goats and 3 in sheep are reported from the semiarid region of northeastern Brazil. Animals of different ages were affected in various seasons of the year. In 5 outbreaks the animals were supplemented with concentrate ration and in 5 others they were only grazing on pastures. In one outbreak sheep were supplemented with an energy-protein-mineral mixture containing 1.3% of sulfur flower. Clinical signs were characterized by blindness, depression, head pressing, circling, grinding of the teeth, incoordination, spastic paralysis, ataxia, depression of the palpebral and pupillary reflexes, lateral strabismus, nystagmus, and dilated pupils. Nine affected animals were treated with thiamine and dexamethasone; 7 of them recovered but 2 died. The diagnosis of the disease was based on the recovered animals after treatment and/or on the histologic lesions. The clinical course varied from 2 to 15 days. On three animals post-mortem examination was made. One had herniation of the cerebellum through the Foramen magnum and softening of the cerebral cortex. The cut surface of the cerebral cortex showed cavitation and yellowish discoloration. Another animal had only cerebellar herniation. In a third animal no gross lesions were observed. Histological changes in the 3 animals were laminar necrosis of the cerebral cortex, and in 2 malacia of the thalamus and the rostral colliculi was also observed. In 9 outbreaks the cause of the disease was not determined, but one was probably due to sulfur toxicosis caused by the high sulfur content of the energy-protein-mineral mixture containing 1.3% of sulfur flower (96% sulfur) and 30% chicken litter (0.39% sulfur).

• Polioencephalomalacia cortical necrosis goats sheep sulfur poisoning thiamine deficiency.

Abstract in Portuguese:

Lima E.F., Riet-Correa F., Tabosa I.M., Dantas A.F.M., Medeiros J.M. & Sucupira Júnior G. 2005. [Polioencephalomalacia in goats and sheep in the semiarid region of northeastern Brazil.] Polioencefalomalacia em caprinos e ovinos na região semi-árida do Nordeste do Brasil. Pesquisa Veterinária Brasileira 25(1):9-14. Centro de Saúde e Tecnologia Rural, Universidade Federal de Campina Grande, Campus de Patos, 58700-000 Patos, PB, Brazil. E-mail: riet@cstr.ufcg.edu.br Seven outbreaks of polioencephalomalacia in goats and 3 in sheep are reported from the semiarid region of northeastern Brazil. Animals of different ages were affected in various seasons of the year. In 5 outbreaks the animals were supplemented with concentrate ration and in 5 others they were only grazing on pastures. In one outbreak sheep were supplemented with an energy-protein-mineral mixture containing 1.3% of sulfur flower. Clinical signs were characterized by blindness, depression, head pressing, circling, grinding of the teeth, incoordination, spastic paralysis, ataxia, depression of the palpebral and pupillary reflexes, lateral strabismus, nystagmus, and dilated pupils. Nine affected animals were treated with thiamine and dexamethasone; 7 of them recovered but 2 died. The diagnosis of the disease was based on the recovered animals after treatment and/or on the histologic lesions. The clinical course varied from 2 to 15 days. On three animals post-mortem examination was made. One had herniation of the cerebellum through the Foramen magnum and softening of the cerebral cortex. The cut surface of the cerebral cortex showed cavitation and yellowish discoloration. Another animal had only cerebellar herniation. In a third animal no gross lesions were observed. Histological changes in the 3 animals were laminar necrosis of the cerebral cortex, and in 2 malacia of the thalamus and the rostral colliculi was also observed. In 9 outbreaks the cause of the disease was not determined, but one was probably due to sulfur toxicosis caused by the high sulfur content of the energy-protein-mineral mixture containing 1.3% of sulfur flower (96% sulfur) and 30% chicken litter (0.39% sulfur).

Abstract in English:

Lima E.S.C., Pinto P.S.A., Santos J.L., Vanetti M.C.D., Bevilacqua P.D., Almeida L.P., Pinto M.S. & Dias F.S. 2004. [Isolation of Salmonella sp and Staphylococcus aureus at swine slaughtering as subsidy for HACCP, the Hazard Analysis and Critical Control Point system.] Isolamento de Salmonella sp e Staphylococcus aureus no processo do abate suíno como subsídio ao sistema de Análise de Perigos e Pontos Críticos de Controle - APPCC. Pesquisa Veterinária Brasileira 24(4):185-190. Depto Veterinária, Universidade Federal de Viçosa, 36571-000 Viçosa, Minas Gerais, Brazil. E-mail: pintopsa@ufv.br This study was done to evaluate the superficial contamination of swine carcasses by Salmonella sp and Staphylococcus aureus, the identification of microbiological hazards in different segments of the processing line, and critical control points (CCPs), through the quantification of risks. A total of 120 surface swabbing carcasses were collected in a slaughterhouse: after the scalding/dehairing (point A), before evisceration (B), after evisceration and splitting (C), and after 24 hours of refrigeration (D). Salmonella sp and S. aureus were isolated from 14 (11.7%) carcasses. No statistical difference between the points studied was observed. The number of S. aureus isolated was between 1.2 and 1.5 log UFC/cm2. It was concluded that the risks observed were the same for both microorganisms.

• Salmonella sp Staphylococcus aureus swine carcass slaughterhouse HACCP.

Abstract in Portuguese:

Lima E.S.C., Pinto P.S.A., Santos J.L., Vanetti M.C.D., Bevilacqua P.D., Almeida L.P., Pinto M.S. & Dias F.S. 2004. [Isolation of Salmonella sp and Staphylococcus aureus at swine slaughtering as subsidy for HACCP, the Hazard Analysis and Critical Control Point system.] Isolamento de Salmonella sp e Staphylococcus aureus no processo do abate suíno como subsídio ao sistema de Análise de Perigos e Pontos Críticos de Controle - APPCC. Pesquisa Veterinária Brasileira 24(4):185-190. Depto Veterinária, Universidade Federal de Viçosa, 36571-000 Viçosa, Minas Gerais, Brazil. E-mail: pintopsa@ufv.br This study was done to evaluate the superficial contamination of swine carcasses by Salmonella sp and Staphylococcus aureus, the identification of microbiological hazards in different segments of the processing line, and critical control points (CCPs), through the quantification of risks. A total of 120 surface swabbing carcasses were collected in a slaughterhouse: after the scalding/dehairing (point A), before evisceration (B), after evisceration and splitting (C), and after 24 hours of refrigeration (D). Salmonella sp and S. aureus were isolated from 14 (11.7%) carcasses. No statistical difference between the points studied was observed. The number of S. aureus isolated was between 1.2 and 1.5 log UFC/cm2. It was concluded that the risks observed were the same for both microorganisms.

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: Guimarães A.M., Lima J.D. & Ribeiro M.EB. 2003. Ultrastructure of Babesia equitrophozoites isolated in Minas Gerais, Brazil. Pesquisa Veterinária Brasileira 23(3):101-104. [Ultra-estrutura de trofozoítos de Babesia equi isolados em Minas Gerais, Brasil.] Departamento de Medicina Veterinária, Universidade Federal de Lavras, Cx. Postal 37, Lavras, MG 37200-000, Brazil. E-mail: amg@ufla.br A transmission electron microscope study was carried out on Babesia equi obtained from a splenectomized horse, from the municipality of Santa Luzia, Minas Gerais, Brazil. The isolate was inoculated into two splenectomized foals (1.05 x 1010 parasitized erythrocytes by B. equi). Trophozoites have a single membrane in direct contact with the cytoplasm of the red blood cells, a prominent nucleus, well-developed rough and smooth endoplasmic reticulum, numerous free ribosomes and small food vacuóles. B. equi trophozoites have a cytostome and a long tubular feeding structure in direct contact with the blood plasmá.

• Babesia equi ultra-estrutura trofozoítos mecanismos de alimentação ultrastructure trophozoites feeding mechanism.

Abstract in Portuguese:

RESUMO: Guimarães A.M., Lima J.D. & Ribeiro M.EB. 2003. Ultrastructure of Babesia equitrophozoites isolated in Minas Gerais, Brazil. Pesquisa Veterinária Brasileira 23(3):101-104. [Ultra-estrutura de trofozoítos de Babesia equi isolados em Minas Gerais, Brasil.] Departamento de Medicina Veterinária, Universidade Federal de Lavras, Cx. Postal 37, Lavras, MG 37200-000, Brazil. E-mail: amg@ufla.br Neste estudo de microsco-pia eletrônica de transmissão utilizou-se um isolado de Babesia equi obtido a partir de um eqüino esplenectomizado, oriundo do município de Santa Luzia, Minas Gerais, Brasil. O isolado foi inoculado em dois potros esplenectomizados (1,05 x 1010 eritrócitos parasitados com B. equi). Os trofozoítos apresentaram uma membrana simples em· contato direto com o citoplasma das· hemácias, núcleo proeminente, retículo endoplasmático liso e rugoso bem desenvolvidos, numerosos ribosomos livres e pequenos vacúolos alimentares. Em trofozoítos de B. equi foi observado citostoma e uma longa estrutura tubular de alimentação em contato direto com o plasma sangüíneo.

Abstract in English:

ABSTRACT.- Corbellini L.G., Pescador C.A., Frantz F.J., Lima M., Ferreiro L. & Driemeier D. 2003. [Abortus by Aspergillus fumigatus and Aspergillus niger in cattle in southern Brazil.] Aborto por Aspergillusfumigatus eAspergillus niger em bovinos no sul do Brasil. Pesquisa Veterinária Brasileira 23(2):82-86. Departamento de Patologia Clínica Veterinária UFRGS, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail:davetpat@vortex.ufrgs.br Mycotic infection has worldwide distribution and may cause placentitis and abortion in almost ali domestic animais. From September 2001 through November 2002 specimens from 147 aborted bovine fetuses along with 34 placentas were submitted to the Laboratory of Veterinary Pathology, Universidade Federal do Rio Grande do Sul, to investigate infectious causes of abortion in southern Brazil. Mycotic abortion was diagnosed in five cases (3.4%). Aspergilfus fumigatus was isolated from four cases and A. niger from one. Virological, bacteriological and direct immunofluorescent antibody tests for Leptospira sp were negative in those cases where Aspergillus sp was cultured. The gestational age of those fetuses ranged from 5 to 8 months. Macroscopic lesions were observed in four cases. One had several Nodular lesions disseminated throughout the liver, two had skin lesions characterized by white-grayish round plaques mostly on the head and neck, and thickened cotyledons in two placentas sent with those fetuses. The histological lesions were observed in the liver, lungs and placenta and consisted primary of necrotizing hepatitis, suppurative bronchopneumonia of varying degrees and necrotizing placentitis. With the use of Grocott's methenamine-silver staining, septate hyphae could be observed in three cases, surrounding necrotizing lesions in the tiver of one fetus and in two placentas. In two cases hyphae were observed in placenta) tissues and not in fetal tissues, indicating the importance of the placenta in diagnosing mycotic abortion in cattle.

• Bovine abortion mycotic abortion Aspergillus spp. Aborto bovino aborto micótico

Abstract in Portuguese:

RESUMO.- Corbellini L.G., Pescador C.A., Frantz F.J., Lima M., Ferreiro L. & Driemeier D. 2003. [Abortus by Aspergillus fumigatus and Aspergillus niger in cattle in southern Brazil.] Aborto por Aspergillusfumigatus eAspergillus niger em bovinos no sul do Brasil. Pesquisa Veterinária Brasileira 23(2):82-86. Departamento de Patologia Clínica Veterinária UFRGS, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail:davetpat@vortex.ufrgs.br As infecções micóticas apresentam distribuição mundial e podem causar placentite e aborto em diversas espécies de animais. Entre setembro 2001 e novembro 2002, foram processados no Setor de Patologia Veterinária, Universidade Federal do Rio Grande do Sul, 147 fetos bovinos abortados com o objetivo de avaliar as principais causas de aborto infeccioso bovino no sul do Brasil. Destes, 34 estavam acompanhados da placenta. Aborto micótico foi diagnosticado em cinco casos (3,4%) mediante cultivo puro de quatro amostras de Aspergillus fumigatus e uma de A. niger associado a lesões histológicas características de fungo. Os exames virológico, bacteriológico e imunofluorescência direta para Leptospira sp foram negativos nestes casos. A idade dos fetos variava entre 5 e 8 meses de idade. Lesões macroscópicas foram observadas em quatro casos e eram caracterizadas por áreas circulares multifocais branco-acinzentadas na pele, principalmente na região da cabeça e dorso, em dois fetos, lesões nodulares no fígado em um caso e espessamento dos cotilédones em duas placentas enviadas juntamente com os fetos. Lesões histológicas foram observadas principalmente no fígado, pulmão e placenta, caracterizadas por hepatite necrótica multifocal, broncopneumonia supurativa e placentite necrótica. Através da coloração de Grocott hifas septadas foram observadas em duas placentas e nas bordas das lesões necróticas no fígado de um feto. Em dois casos hifas foram observadas somente na placenta e não no feto, salientando-se a importância deste tecido para o diagnóstico de aborto micótico bovino.

Abstract in English:

ABSTRACT.- Franco A.C., Spilki F.R., Esteves P.A., Lima M., Weiblen R., Flores E.F., Rijsewijk F.A.M. & Roehe P.M. 2002. A Brazilian glycoprotein E-negative bovine herpesvirus type 1.2a (BHV-1.2a) mutant is attenuated for cattle and induces protection against wild-type virus challenge. Pesquisa Veterinária Brasileira 22(4):135-140. [Um mutante gE-negativo de herpesvírus bovino tipo 1.2a é atenuado para bovinos e induz proteção frente ao desafio com vírus de campo.] Centro de Pesquisas Veterinárias Desidério Finamor, Fepagro-Saúde Animal, Cx. Postal 2076, Porto Alegre, RS 90001-970, Brazil. E-mail: proehe@ufrgs.br The authors previously reported the construction of a glycoprotein E-deleted (gE·) mutante of bovine herpesvirus type 1.2a (BHV-1.2a). This mutant, 265gE·, was designed as a vacinal strain for differential vaccines, allowing the distinction between vaccinated and naturally infected cattle. In order to determine the safety and efficacy of this candidate vaccine virus, a group of calves was inoculated with 265gE·. The virus was detected in secretions of inoculated calves to lower titres and for a shorter period than the parental virus inoculated in control calves. Twenty one days after inoculation, the calves were challenged with the wild type parental virus. Only mild signs of infection were detected on vaccinated calves, whereas nonvaccinated controls displayed intense rhinotracheitis and shed virus for longer and to higher titres than vaccinated calves. Six months after vaccination, both vaccinated and control groups were subjected to reactivation of potentially latent virus. The mutant 265gE· could not be reactivated from vaccinated calves. The clinical signs observed, following the reactivation of the parental virus, were again much milder on vaccinated than on non-vaccinated calves. Moreover, parental vírus shedding was considerably reduced on vaccinated calves at reactivation. In view of its attenuation, immunogenicity and protective effect upon challenge and reactivation with a virulent BHV-1, the mutant 265gE· was shown to be suitable for use as a BHV-1 differential vaccine vírus.

• BHV-1 differential vaccine gE deletion IBR vacina diferencial gE.

Abstract in Portuguese:

RESUMO.- Franco A.C., Spilki F.R., Esteves P.A., Lima M., Weiblen R., Flores E.F., Rijsewijk F.A.M. & Roehe P.M. 2002. A Brazilian glycoprotein E-negative bovine herpesvirus type 1.2a (BHV-1.2a) mutant is attenuated for cattle and induces protection against wild-type virus challenge. Pesquisa Veterinária Brasileira 22(4):135-140. [Um mutante gE-negativo de herpesvírus bovino tipo 1.2a é atenuado para bovinos e induz proteção frente ao desafio com vírus de campo.] Centro de Pesquisas Veterinárias Desidério Finamor, Fepagro-Saúde Animal, Cx. Postal 2076, Porto Alegre, RS 90001-970, Brazil. E-mail: proehe@ufrgs.br Em estudo prévio os autores reportaram a construção de um mutante do Vírus da Rinotraqueíte Infecciosa Bovina (IBR) ou Herpesvírus Bovino tipo 1.2a (BHV-1.2a), do qual foi deletado o gene que codifica a glicoproteina E. Esse mutante (265gE-) foi construído a partir de uma amostra autóctone do vírus, tendo como objetivo seu uso como amostra vacinai em vacinas diferenciais, capazes de permitir a diferenciação entre animais vacinados e infectados com vírus de campo. Para determinar a atenuação e eficácia do 265gE· como imunógeno, bezerros foram inoculados por via intranasal com 106,9 DICC50 do mesmo. O vírus foi detectado em secreções dos animais inoculados em títulos mais baixos e por um período mais curto do que a amostra virulenta parental, inoculada em animais controle. Vinte e um dias após, os animais inoculados com o vírus mutante foram desafiados com a amostra parental, apresentando somente sinais leves de infecção. Os animais controle apresentaram intensa rinotraqueíte e excretaram vírus em títulos mais elevados e por mais tempo do que os vacinados. Seis meses após a vacinação, foi examinada a capacidade de reativação da infecção nos bezerros, através da administração de corticosteróides. O mutante 265gE- não foi reativado dos animais vàcinados. Os sinais clínicos consequentes à reativação do vírus parental foram muito atenuados nos animais vacinados, em comparação com os não vacinados. Além disso, a excreção de vírus de campo foi consideravelmente reduzida nestes últimos. Em vista de sua atenuação, imunogenicidade e efeito protetivo frente ao desafio com uma amostra virulenta de BHV-1 e subseqüente reativação, o mutante 265gE- demonstrou apresentar grande potencial para ser utilizado como vírus vacinai em vacinas diferenciais contra o BHV-1.

Abstract in English:

ABSTRACT.- Molnár L., Molnár E., Lima E.S.C. & Dias H.L.T. 2002. [Evaluation of six serological tests for the diagnosis of brucellosis in water buffaloes.] Avaliação de seis testes sorológicos no diagnóstico da brucelose bubalina. Pesquisa Veterinária Brasileira 22(2):41-44. Centro Agropecuário, Lidea, Universidade Federal do Pará, Belém, PA 66075-900, Brazil. Four hundred and forty buffalo sera, selected from about 1,200 blood samples of another study, were examined. The samples were tested by six serological methods: two of agglutination, two of indirect ELISA and two of competitive ELISA. To determine the relative sensitivity and specificity of different tests, animals with a positive result to competitive ELISA of the FAO/IAEA were considered as infected. The relative sensitivity of competitive ELISA, indirect ELISA with conjugate anti-bovine light chain monoclonal antibody labelled with HRPO, indirect ELISA with anti-bovine lgG conjugate, rose Bengal test and rapid slide agglutination test was 100%, 98.57%, 97.14%, 91.42% and 79.28%, and the relative specificity 99.33%, 97.33%, 95.66%, 94.00% and 86.33%, respectively. The value of the different serological tests for the diagnosis of brucellosis is discussed.

• Brucellosis water buffalo competitive ELISA indirect ELISA agglutination Bucelose bubalino ELISA competitivo ELISA indireto aglutinação.

Abstract in Portuguese:

RESUMO.- Molnár L., Molnár E., Lima E.S.C. & Dias H.L.T. 2002. [Evaluation of six serological tests for the diagnosis of brucellosis in water buffaloes.] Avaliação de seis testes sorológicos no diagnóstico da brucelose bubalina. Pesquisa Veterinária Brasileira 22(2):41-44. Centro Agropecuário, Lidea, Universidade Federal do Pará, Belém, PA 66075-900, Brazil. Foram examinados 440 soros bubalinos, selecionados em um outro exame de cerca de 1200 amostras sangüíneas. Utilizaram-se seis diferentes testes sorológicos para o exame dessas amostras: dois de aglutinação, dois de ELISA indireto e dois de ELISA competitivo. Os animais positivos no ELISA competitivo da FAO/IAEA foram considerados como infectados, e a comparação com os resultados dos outros testes aconteceu neste sentido. A sensibilidade relativa foi de 100%, 98,57%, 97,14%, 91,42% e 79,28%, e a especificidade relativa de 99,33%, 97,33%, 95,66%, 94,00% e 86,33% nas provas de ELISA competitivo, ELISA indireto com conjugado antibovino de cadeia leve (anticorpo monoclonal com HRPO), ELISA indireto com conjugado contra lgG bovino total, teste do antígeno acidificado tamponado e aglutinação rápida, respectivamente. Discute-se o valor dos diferentes testes sorológicos no diagnóstico da brucelose.

Abstract in English:

ABSTRACT.- Scherer C.F.C., Flores E.F., Weiblen R., Kreutz L.C., Dürr J.W., Brum L.P., Quadros V.L. & Lima M. 2000. [A rapid virus-neutralization test for detection of antibodies against bovine viral diarrhea virus (BVDV) in milk.] Técnica rápida de neutralização viral para pesquisa de anticorpos contra o vírus da Diarréia Viral Bovina (BVDV) no leite. Pesquisa Veterinária Brasileira 22(2):45-50. Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. The identification of bovine viral diarrhea virus (BVDV) positive herds through detection of antibodies in milk may viabilize large scale control/eradication programs. With this objective, the virus neutralization test (VN) was adapted to detect BVDV antibodies in milk. The adaptation consisted of a reduction in the time of incubation followed by detection of viral antigens in the indicator cells by immunofluorescence (IFA) and allowed readings at 24 hours. The rapid virus neutralization test (RVN) was initially tested in 1,335 serum samples, showing a 93. 7% sensitivity and 91.1 % agreement with the traditional VN. The RVN was also used to test 423 bovine sera that were toxic for cell culture in the traditional VN test, detecting 316 (74.7%) positive samples. Testing of matched serum and milk samples from BVDV seropositive cows showed that the VNR can detect antibodies in the milk of cows with serum neutralizing titers as low as 10. Anti-BVDV neutralizing activity was detected in milk of 97.4% (191/196) of cows with serum titers 3320; in 92.9% (79/85) of cows with titers of 160; in 88% (59/67) of cows with serum titers of 80. The frequency of BVDV antibodies in milk was 76.9% (40/52) for cows with serum titers of 40; 61.3% (19/31) for cows with titers of 20 and 33.3% (10/30) for cows with serum titers of 20. These results demonstrate that the RVN test is adequate for detecting BVDV antibodies in milk, mainly in cows having moderate to high serum titers, and therefore may be used for testing bulk milk samples to identify herds with viral activity. The use of this test may viabilize large scale programs for control/eradication of BVDV infection. It allows to assay a large number of samples and identify positive herds through testing milk routinely submitted for somatic cell counts (SCC), reducing costs with individual sample collection, shipping and testing.

• Bovine viral diarrhea virus BVDV diagnosis antibodies milk Vírus da Diarréia Viral Bovina diagnóstico leite anticorpos.

Abstract in Portuguese:

RESUMO.- Scherer C.F.C., Flores E.F., Weiblen R., Kreutz L.C., Dürr J.W., Brum L.P., Quadros V.L. & Lima M. 2000. [A rapid virus-neutralization test for detection of antibodies against bovine viral diarrhea virus (BVDV) in milk.] Técnica rápida de neutralização viral para pesquisa de anticorpos contra o vírus da Diarréia Viral Bovina (BVDV) no leite. Pesquisa Veterinária Brasileira 22(2):45-50. Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. A identificação de rebanhos positivos para o vírus da Diarréia Viral Bovina (BVDV) através de detecção de anticorpos no leite pode viabilizar programas de controle em larga escala. Com esse objetivo, a técnica de soroneutralização (SN) foi adaptada para a pesquisa de anticorpos em amostras de leite. A adaptação consistiu na redução do tempo de incubação do teste, seguida da detecção de antígenos virais por imunofluorescência. A redução do tempo de incubação minimizou os efeitos tóxicos do leite sobre as células de cultivo, além de permitir a obtenção dos resultados em 24 horas. A técnica rápida (SNR) foi inicialmente testada em 1.335 amostras de soro bovino, apresentando sensibilidade de 93,7% e concordância de 91, 1% em relação à SN tradicional. A SNR foi também utilizada para testar 423 amostras de soro bovino que apresentaram toxicidade para as células na SN tradicional, detectando 316 (74,7%) amostras positivas. O teste de amostras de soro e leite de 520 vacas em lactação demonstrou que a SNR pode detectar anticorpos no leite de vacas com títulos séricos a partir de 10. Atividade neutralizante anti-BVDV no leite foi detectada em 97,4% (191/196) de vacas com títulos séricos 3 320; em 92,9% (79/85) de vacas com títulos de 160; em 88% (59/67) de vacas títulos de 80. A freqüência de animais positivos na SNR foi de 76,9% (40/52) para animais com títulos séricos de 40; 61,3% (19/31) com títulos de 20 e de 33,3% (10/30) para vacas com títulos de 10. Esses resultados demonstram que a técnica de SNR é adequada para a pesquisa de anticorpos anti-BVDV no leite, principalmente em animais com títulos moderados e altos de anticorpos. Essa técnica pode ser utilizada para testar amostras coletivas de leite e identificar rebanhos com atividade viral. A utilização dessa técnica pode viabilizar programas regionais de combate à infecção, pois permite testar um grande número de amostras e identificar rebanhos positivos através do leite enviado rotineiramente para contagem de células somáticas (CCS), reduzindo significativamente os custos com a coleta individual, transporte e teste de amostras.

Abstract in English:

ABSTRACT.- Spilki F.R., Esteves P.A., Franco A.C., Lima M., Holz C.L., Batista H.B.C.R., Driemeier D., Flores E.F., Weiblen R. & Roehe P.M. 2002. [Neurovirulence and neuroinvasiveness of bovine herpesvirus type 1 and 5 in rabbits.] Neurovirulência e neuroinvasividade de Herpesvírus bovinos tipos 1 e 5 em coelhos. Pesquisa Veterinária Brasileira 22(2):58-63. Centro de Pesquisas Veterinárias Desidério Finamor, Estrada do Conde 6000, Cx. Postal 47, Eldorado do Sul, RS 92990-000, Brazil. E-mail: proehe@orion.ufrgs.br In order to determine the capacity of bovine herpesvirus type 1 and 5 (BHV-1 and BHV-5) to invade, multiply and spread along the central nervous system (CNS) (neuroinvasiveness), as well as their potential to induce neurological illness (neurovirulence), 30 to 35 days old rabbits were inoculated with the BHV-5 strain EVI 88 / 95 and Los Angeles and Cooper BHV-1 strains, by the intrathecal (IT) and intranasal (IN) routes. The BHV-5 strain induced severe neurological clinical signs in 100% (12/12) of the rabbits inoculated by both routes. Histopathological examination revealed multifocal non-suppurative meningoencephalitis, characterized by multifocal gliosis and perivascular cuffing. Virus was recovered from many parts of the brain. Both BHV-1 strains, when inoculated via 1T route, were not neurovirulent. The strain Los Angeles, after IN inoculation, induced signs of severe respiratory disease (7/7), as well as signs of neurological impairment, indistinguishable from those induced by BHV-5, in 57% (4/7) of the infected rabbits. However, the rabbits with nervous signs revealed at histopathology vasculitis and thrombosis in lungs and brain, the latter with foci of neuronal necrosis, but no lesions indicative of encephalitis, suggesting that neural damage was probably consequent to tissue anoxia. The BHV-1 strain Cooper, after IN inoculation, induced only mild signs of respiratory disease. These findings indicate that the BHV-5 strain was both neuroinvasive and neurovirulent, since it was capable of invading, spreading and multiplying in the rabbits brains by both routes of inoculation, yet causing neurological disease, apparently consequent to vírus induced neural damage. The BHV-1 Los Angeles strain was not neuroinvasive, whereas its neurovirulence was probably consequent to tissue anoxia, which histologically seemed not to be related to direct viral pathogenic effect. The BHV-1 strain Cooper was neither neurovirulent nor neuroinvasive for rabbits. It is possible that these observations bear relationship with the frequent association of BHV-5 with encephalitis in cattle, as opposed to BHV-1 encephalitis, which is a rare event in nature.

• Infectious bovine rhinotracheitis IBR BHV-1 bovine encephalitis herpesvirus BHV-5 Rinotraqueíte infecciosa bovina herpesvírus da encefalite bovina.

Abstract in Portuguese:

RESUMO.- Spilki F.R., Esteves P.A., Franco A.C., Lima M., Holz C.L., Batista H.B.C.R., Driemeier D., Flores E.F., Weiblen R. & Roehe P.M. 2002. [Neurovirulence and neuroinvasiveness of bovine herpesvirus type 1 and 5 in rabbits.] Neurovirulência e neuroinvasividade de Herpesvírus bovinos tipos 1 e 5 em coelhos. Pesquisa Veterinária Brasileira 22(2):58-63. Centro de Pesquisas Veterinárias Desidério Finamor, Estrada do Conde 6000, Cx. Postal 47, Eldorado do Sul, RS 92990-000, Brazil. E-mail: proehe@orion.ufrgs.br Com o objetivo de avaliar a capacidade dos herpesvírus bovinos tipos 1 e 5 (BHV-1 e BHV-5) de invadir e replicar no sistema nervoso central (SNC) (neuroinvasividade), bem como sua capacidade de induzir doença neurológica (neurovirulência), coelhos com 30 a 35 dias de idade foram inoculados com uma amostra do Herpesvírus da Encefalite Bovina (BHV-5; amostra EVI 88/95) ou com amostras de BHV-1 (Los Angeles ou Cooper), pelas vias intratecal (IT) e intranasal (IN). A inoculação da amostra de BHV-5, tanto pela via 1T como IN, induziu sinais clínicos neurológicos em 100% (12/12) dos coelhos inoculados. Os exames histopatológicos revelaram um quadro de meningoencefalite não-purulenta multifocal, caracterizada por gliose multifocal e infiltrados perivasculares. O vírus foi isolado de várias áreas do SNC desses animais. As amostras de BHV-1, quando inoculadas pela via IT, não foram neurovirulentas. A amostra Los Angeles de BHV-1, quando administrada pela via IN, induziu sinais respiratórios severos, além de sinais neurológicos em 57% (4/7) dos animais inoculados. Entretanto, o exame histopatológico destes quatro animais revelou vasculite e trombose no pulmão e cérebro, este último apresentando focos de necrose neuronal, porém sem lesões indicativas de encefalite. Isso sugere que os sinais neurológicos foram, provavelmente, consequentes a prejuízos no fluxo sangüíneo encefálico, e não a danos neuronais provocados pela inoculação desse vírus. A amostra Cooper de BHV-1, quando inoculada pela via IN, induziu apenas sinais leves de infecção respiratória. Estes resultados indicam que apenas a amostra de BHV-5 foi capaz de invadir e replicar no encéfalo dos coelhos quando inoculada tanto por via IN como IT, apresentando neuroinvasividade e neurovirulência. É possível que estas observações tenham relação com o fato de amostras de BHV-5 freqüentemente causarem encefalites, em contraposição a infecções pelo BHV-1, onde encefalites são raramente observadas.