PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

Garmatz S.L., Irigoyen L.F., Rech R.R., Brown C.C., Zhang J. & Barros C.S.L. 2004. [Malignant catarrhal fever in cattle in Rio Grande do Sul, Brazil: Experimental transmission to cattle and characterization of the etiological agent.] Febre catarral maligna em bovinos no Rio Grande do Sul: transmissão experimental para bovinos e caracterização do agente etiológico. Pesquisa Veterinária Brasileira 24(2):93-106. Depto Patologia, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. E-mail: claudioslbarros@uol.com.br Two oubreaks of malignant catarrhal fever (MCF) occurring in cattle on two farms (A and B) in the municipality of Santiago, state of Rio Grande do Sul (RS), Brazil, and the transmission of the disease to susceptible calves as well as the detection of ovine herpesvirus-2 (OvHV-2) in tissues of affected cattle are reported. The two epizootics occurred from November 2001 to February 2002 (Farm A) and in January-February 2003 (Farm B). Numbers of cattle at risk, morbidity and letality rates were respectively 170, 10.59% and 83.33% for Farm A and 500, 2.4% and 100% for Farm B. Contact between affected cattle and sheep was detected in both farms, but lambing ewes were present only in farm A. Duration of clinical courses, gross findings and histopathology were the same for the affected cattle in both farms. Most affected cattle died or were euthanatized in extremis after a clinical course of 2-8 days. Clinical signs included fever (40.5 and 41.5°C), nasal and ocular discharge, corneal opacity, conjunctivitis, drooling, erosions and ulcerations of the mucosae, diarrhea, hematuria, and neurological disturbances. Eleven necropsies (9 on Farm A, 2 on Farm B) were performed. Gross lesions included erosions and ulcers affecting the mucosae of nasal turbinates, oral cavity, gastrointestinal and urogenital tracts; hemorrhage and necrosis of the tip of the buccal papillae, lymph node enlargement, multifocal white foci in renal cortex, and hyperemia of leptomeninges. Microscopically, there were arteritis and fibrinoid degeneration in medium and small arteries and arterioles of multiple organs and tissues, necrosis and inflammation in several mucosal surfaces, keratitis, conjunctivitis, uveitis, intersticial nephritis, and encephalitis. Transmission experiments were attempted in five calves (E1-E5) by inocculating each of them intravenously with 500 ml of whole heparinized blood from a MCF affected cow. The transmission was suscessful in at least three (E1-E3) of the experimental calves which became sick after an incubation period of 15-27 days. Four experimental calves either died or were euthanatized in extremis after a clinical course which varied from 3 days to 8 weeks. The remaining experimental calf (E5) recovered from a mild disease and was euthanatized 14 weeks after inocculation. Necropsies were performed in all five calves. Clinical signs, necropsy and histopathological findings of three calves (E1-E3) were characteristic of MCF. OvHV-2 viral DNA was detected by the polimerase chain reaction (PCR) test in paraffin embedded tissues from seven cattle out of the 11 spontaneous MCF cases and from three experimental calves (E1-E3). PCR tests resulted negative in the remaining four of the 11 spontaneous MCF cases tested and in two (E4,E5) of the five experimental calves. Immunohistochemistry performed in sections of lymphoid tissue from calf E4 failed to detect BVD virus antigen. The experimental transmission of MCF and the characterization of the etiological agent as OvHV-2 were successfully attempted in cattle for the first time in Brazil.

• Infectious diseases malignant catarrhal fever viral diseases diseases of cattle pathology polimerase chain reation.

Abstract in Portuguese:

Garmatz S.L., Irigoyen L.F., Rech R.R., Brown C.C., Zhang J. & Barros C.S.L. 2004. [Malignant catarrhal fever in cattle in Rio Grande do Sul, Brazil: Experimental transmission to cattle and characterization of the etiological agent.] Febre catarral maligna em bovinos no Rio Grande do Sul: transmissão experimental para bovinos e caracterização do agente etiológico. Pesquisa Veterinária Brasileira 24(2):93-106. Depto Patologia, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. E-mail: claudioslbarros@uol.com.br Two oubreaks of malignant catarrhal fever (MCF) occurring in cattle on two farms (A and B) in the municipality of Santiago, state of Rio Grande do Sul (RS), Brazil, and the transmission of the disease to susceptible calves as well as the detection of ovine herpesvirus-2 (OvHV-2) in tissues of affected cattle are reported. The two epizootics occurred from November 2001 to February 2002 (Farm A) and in January-February 2003 (Farm B). Numbers of cattle at risk, morbidity and letality rates were respectively 170, 10.59% and 83.33% for Farm A and 500, 2.4% and 100% for Farm B. Contact between affected cattle and sheep was detected in both farms, but lambing ewes were present only in farm A. Duration of clinical courses, gross findings and histopathology were the same for the affected cattle in both farms. Most affected cattle died or were euthanatized in extremis after a clinical course of 2-8 days. Clinical signs included fever (40.5 and 41.5°C), nasal and ocular discharge, corneal opacity, conjunctivitis, drooling, erosions and ulcerations of the mucosae, diarrhea, hematuria, and neurological disturbances. Eleven necropsies (9 on Farm A, 2 on Farm B) were performed. Gross lesions included erosions and ulcers affecting the mucosae of nasal turbinates, oral cavity, gastrointestinal and urogenital tracts; hemorrhage and necrosis of the tip of the buccal papillae, lymph node enlargement, multifocal white foci in renal cortex, and hyperemia of leptomeninges. Microscopically, there were arteritis and fibrinoid degeneration in medium and small arteries and arterioles of multiple organs and tissues, necrosis and inflammation in several mucosal surfaces, keratitis, conjunctivitis, uveitis, intersticial nephritis, and encephalitis. Transmission experiments were attempted in five calves (E1-E5) by inocculating each of them intravenously with 500 ml of whole heparinized blood from a MCF affected cow. The transmission was suscessful in at least three (E1-E3) of the experimental calves which became sick after an incubation period of 15-27 days. Four experimental calves either died or were euthanatized in extremis after a clinical course which varied from 3 days to 8 weeks. The remaining experimental calf (E5) recovered from a mild disease and was euthanatized 14 weeks after inocculation. Necropsies were performed in all five calves. Clinical signs, necropsy and histopathological findings of three calves (E1-E3) were characteristic of MCF. OvHV-2 viral DNA was detected by the polimerase chain reaction (PCR) test in paraffin embedded tissues from seven cattle out of the 11 spontaneous MCF cases and from three experimental calves (E1-E3). PCR tests resulted negative in the remaining four of the 11 spontaneous MCF cases tested and in two (E4,E5) of the five experimental calves. Immunohistochemistry performed in sections of lymphoid tissue from calf E4 failed to detect BVD virus antigen. The experimental transmission of MCF and the characterization of the etiological agent as OvHV-2 were successfully attempted in cattle for the first time in Brazil.

Abstract in English:

Sallis E.S.V., Raffi M.B. & Riet-Correa F. 2004. [Experimental poisoning in sheep with frozen or dried Ramaria flavo-brunnescens.] Intoxicação experimental em ovinos com Ramaria flavo-brunnescens congelada ou dessecada. Pesquisa Veterinária Brasileira 24(2):107-110. Faculdade de Veterinária, Universidade Federal de Pelotas, Campus Universitário, Cx. Postal 354, Pelotas, RS 96010-900, Brazil. E-mail: esvsallis@yahoo.com.br Ramaria flavo-brunnescens is a toxic mushroom affecting cattle and sheep. Its active principle is unknown. The experiment was done to obtain information about the toxicity of the mushroom after been frozen or dried. R. flavo-brunnescens was collected in autumn 1991. One part was frozen at –15ºC for 2-4 months, and another was dried in the shade. Other samples collected at the same time were given immediately after harvest to two sheep, at a total dose of 200 g/kg for 3 and 4 days, causing severe poisoning. The dried mushroom did not cause clinical signs at doses of 60 and 75 g/kg bw (equivalent to 400 and 500 g/kg of the fresh mushroom). The frozen mushroom at 200 g/kg bw caused hyperthermia, depression, hyperemia of the sclera and hemorrhages of the anterior chamber of the eye. Sheep that ingested 350 and 400 g/kg bw showed also nervous signs, and with the highest dose hyperemic lesions of the coronary band. All sheep recovered within 3 to 12 days. Clinical signs when given the frozen mushroom were less severe than clinical signs induced by the fresh mushroom. These results showed loss of toxicity of the dried material and decrease of toxicity of the frozen mushroom. It is suggested that chemical analyses for the toxic principle can be performed with the frozen or lyophilized mushroom.

• Poisonous plants toxic mushroom Ramaria flavo-brunnescens sheep.

Abstract in Portuguese:

Sallis E.S.V., Raffi M.B. & Riet-Correa F. 2004. [Experimental poisoning in sheep with frozen or dried Ramaria flavo-brunnescens.] Intoxicação experimental em ovinos com Ramaria flavo-brunnescens congelada ou dessecada. Pesquisa Veterinária Brasileira 24(2):107-110. Faculdade de Veterinária, Universidade Federal de Pelotas, Campus Universitário, Cx. Postal 354, Pelotas, RS 96010-900, Brazil. E-mail: esvsallis@yahoo.com.br Ramaria flavo-brunnescens is a toxic mushroom affecting cattle and sheep. Its active principle is unknown. The experiment was done to obtain information about the toxicity of the mushroom after been frozen or dried. R. flavo-brunnescens was collected in autumn 1991. One part was frozen at –15ºC for 2-4 months, and another was dried in the shade. Other samples collected at the same time were given immediately after harvest to two sheep, at a total dose of 200 g/kg for 3 and 4 days, causing severe poisoning. The dried mushroom did not cause clinical signs at doses of 60 and 75 g/kg bw (equivalent to 400 and 500 g/kg of the fresh mushroom). The frozen mushroom at 200 g/kg bw caused hyperthermia, depression, hyperemia of the sclera and hemorrhages of the anterior chamber of the eye. Sheep that ingested 350 and 400 g/kg bw showed also nervous signs, and with the highest dose hyperemic lesions of the coronary band. All sheep recovered within 3 to 12 days. Clinical signs when given the frozen mushroom were less severe than clinical signs induced by the fresh mushroom. These results showed loss of toxicity of the dried material and decrease of toxicity of the frozen mushroom. It is suggested that chemical analyses for the toxic principle can be performed with the frozen or lyophilized mushroom.

Abstract in English:

Pereira M.S.V., Siqueira Júnior J.P. & Campos Takaki G.M. 2004. [Elimination of resistance to drugs by fluoroquinolones in bovine strains of Staphylococcus aureus.] Eliminação de resistência a drogas por fluorquinolonas em Staphylococcus aureus de origem bovina. Pesquisa Veterinária Brasileira 24(1):11-14. Depto Biologia Molecular, Centro de Ciências Exatas e da Natureza, Universidade Federal da Paraiba, João Pessoa, PB 58059-900, Brazil. E-mail: svieira@dbm.ufpb.br Bovine strains of Staphylococcus aureus were submitted to treatment with four fluoro-quinolones in subinhibitory concentrations (1/2 x MICs) to evaluate their influence on the curing of plasmids. Ciprofloxacin showed to be the most efficient by eliminating resistance to streptomycin, tetracyclin, penicillin, and cadmium nitrate. Norfloxacin and pefloxacin eliminated penicillin- and tetracyclin-resistance respectively. Otherwise, plasmids elimination by ofloxacin was not evidenced. The results obtained in this study confirm the potential of fluoroquinolones to eliminate antibiotic-resistant plasmids, and showed to be a valuable contribution for the prevention of multi-resistant strains, and may even enhance their sensitivity to other chemotherapeutic agents.

• Staphylococcus aureus fluoroquinolones elimination of resistance to drugs.

Abstract in Portuguese:

Pereira M.S.V., Siqueira Júnior J.P. & Campos Takaki G.M. 2004. [Elimination of resistance to drugs by fluoroquinolones in bovine strains of Staphylococcus aureus.] Eliminação de resistência a drogas por fluorquinolonas em Staphylococcus aureus de origem bovina. Pesquisa Veterinária Brasileira 24(1):11-14. Depto Biologia Molecular, Centro de Ciências Exatas e da Natureza, Universidade Federal da Paraiba, João Pessoa, PB 58059-900, Brazil. E-mail: svieira@dbm.ufpb.br Bovine strains of Staphylococcus aureus were submitted to treatment with four fluoro-quinolones in subinhibitory concentrations (1/2 x MICs) to evaluate their influence on the curing of plasmids. Ciprofloxacin showed to be the most efficient by eliminating resistance to streptomycin, tetracyclin, penicillin, and cadmium nitrate. Norfloxacin and pefloxacin eliminated penicillin- and tetracyclin-resistance respectively. Otherwise, plasmids elimination by ofloxacin was not evidenced. The results obtained in this study confirm the potential of fluoroquinolones to eliminate antibiotic-resistant plasmids, and showed to be a valuable contribution for the prevention of multi-resistant strains, and may even enhance their sensitivity to other chemotherapeutic agents.

Abstract in English:

Malafaia P., Pimentel V.A., Freitas K.P., Coelho C.D., Brito M.F. & Peixoto P.V. 2003. [Daily weight gain, economic, nutritional and clinical aspects of goats receiving two types of mineral supplements.] Desempenho ponderal, aspectos econômicos, nutricionais e clínicos de caprinos submetidos a dois esquemas de suplementação mineral. Pesquisa Veterinária Brasileira 24(1):15-22. Depto de Nutrição Animal e Pastagem, Instituto de Zootecnia, UFRRJ, Km 47, Seropédica, RJ 23835-000, Brazil. E-mail: malafaia1@ig.com.br Daily weight gain, economic, nutritional and clinical aspects of goats receiving two types of mineral supplements were evaluated during 148 days in Seropédica county, Rio de Janeiro, Brazil. The animals were daily fed with fresh and chopped Andropogon gayanus or Pennisetum purpureum; the concentrate ration was formulated without a mineral mixture and was offered together with the forage of 300 up to 500 g/animal/day. Sixteen goats were divided into two groups of 8 animals each (4 small and 4 larger goats). For one group a commercial mineral mixture was used, and to the other group a selective mineral mixture containing only Na, P and Cu was offered. The animals were weighed every 30 days and the mineral supplement intake was measured monthly. Blood samples were collected at the beginning and the end of the experimental period. There was no difference in daily weight gain when the selective or commercial mixture was given for both groups. The selective mineral supplementation was 2.3 (for the smaller animals) and 3.9 (for larger animals) times more economic than the conventional supplementation with a “complete” mineral mix. The two groups showed recovery of hair quality and had an increase of hair pigmentation around the eyes, which was attributed to the correction of copper deficiency. In the larger animals the intake of both supplements increased the red blood cells counts, and the haematocrit and haemoglobin values. The forage intake was sufficient to meet the Ca and Co needs of the animals, and for P the concentrate intake was sufficient to overcome the demand for this element. The requirement of zinc was supplied through the forage and the concentrate ration. Thus, when an adequate concentrate and a good quality forage is given, only the deficient mineral element(s) should be supplemented. In this specific case only Na and Cu should be given. The result of this experiment confirms the hypothesis that selective mineral supplementation – i.e. supplementation of the mineral element(s) deficient in the diet – was correct and can result in reduction of the cost with the mineral supplementation of a herd.

• Goats copper deficiency mineral supplementation tropical forages.

Abstract in Portuguese:

Malafaia P., Pimentel V.A., Freitas K.P., Coelho C.D., Brito M.F. & Peixoto P.V. 2003. [Daily weight gain, economic, nutritional and clinical aspects of goats receiving two types of mineral supplements.] Desempenho ponderal, aspectos econômicos, nutricionais e clínicos de caprinos submetidos a dois esquemas de suplementação mineral. Pesquisa Veterinária Brasileira 24(1):15-22. Depto de Nutrição Animal e Pastagem, Instituto de Zootecnia, UFRRJ, Km 47, Seropédica, RJ 23835-000, Brazil. E-mail: malafaia1@ig.com.br Daily weight gain, economic, nutritional and clinical aspects of goats receiving two types of mineral supplements were evaluated during 148 days in Seropédica county, Rio de Janeiro, Brazil. The animals were daily fed with fresh and chopped Andropogon gayanus or Pennisetum purpureum; the concentrate ration was formulated without a mineral mixture and was offered together with the forage of 300 up to 500 g/animal/day. Sixteen goats were divided into two groups of 8 animals each (4 small and 4 larger goats). For one group a commercial mineral mixture was used, and to the other group a selective mineral mixture containing only Na, P and Cu was offered. The animals were weighed every 30 days and the mineral supplement intake was measured monthly. Blood samples were collected at the beginning and the end of the experimental period. There was no difference in daily weight gain when the selective or commercial mixture was given for both groups. The selective mineral supplementation was 2.3 (for the smaller animals) and 3.9 (for larger animals) times more economic than the conventional supplementation with a “complete” mineral mix. The two groups showed recovery of hair quality and had an increase of hair pigmentation around the eyes, which was attributed to the correction of copper deficiency. In the larger animals the intake of both supplements increased the red blood cells counts, and the haematocrit and haemoglobin values. The forage intake was sufficient to meet the Ca and Co needs of the animals, and for P the concentrate intake was sufficient to overcome the demand for this element. The requirement of zinc was supplied through the forage and the concentrate ration. Thus, when an adequate concentrate and a good quality forage is given, only the deficient mineral element(s) should be supplemented. In this specific case only Na and Cu should be given. The result of this experiment confirms the hypothesis that selective mineral supplementation – i.e. supplementation of the mineral element(s) deficient in the diet – was correct and can result in reduction of the cost with the mineral supplementation of a herd.

Abstract in English:

Cattani C.S.O., Colodel E.M., Traverso S.D., Correa A.M.R., & Driemeier D. 2004. [Experimental poisoning by Dodonea viscosa (Sapindaceae) in cattle.] Intoxicação experimental por Dodonea viscosa (Sapindaceae) em bovinos. Pesquisa Veterinária Brasileira 24(1):31-34. Depto Patologia Clínica Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Bairro Agronomia, Cx. Postal 15094, Porto Alegre, RS 91540-000, Brazil. E-mail: moleta@terra.com.br The leaves of Dodonea viscosa were force fed to five bovines. Four received the leaves in fresh green stage and the fifth dried ones. Clinical signs were observed in four of the bovines that died; the fifth did not show signs of poisoning. The fresh green plant was proved to be toxic from a dose of 25g/kg on. Dried leaves fed at the dose of 30 g/kg were also toxic. All the animals that died showed clinical signs from 13h30min to 45h after the ingestion of the plant and and death followed within about 48h. The clinical course lasted for about 8h30min until death. The main symptoms where apathy, anorexia, slight tenesmus, muscle trembling, difficulties to keep consciousness, pressing the head against obstacles, lateral recumbency, paddling movements, coma and death. The most significant macroscopic alterations were observed in the liver, with accentuation of the lobular pattern, dark-red areas interspersed with yellowish areas. Petechiae were found in serosal membranes of the abdominal and thoracic organs as well as the intestines. The main microscopic change was hepatic centrolobular coagulative necrosis, associated with congestion and hemorrhages.

• Dodonea viscosa diseases of cattle plant poisoning hepatocellular necrosis.

Abstract in Portuguese:

Cattani C.S.O., Colodel E.M., Traverso S.D., Correa A.M.R., & Driemeier D. 2004. [Experimental poisoning by Dodonea viscosa (Sapindaceae) in cattle.] Intoxicação experimental por Dodonea viscosa (Sapindaceae) em bovinos. Pesquisa Veterinária Brasileira 24(1):31-34. Depto Patologia Clínica Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Bairro Agronomia, Cx. Postal 15094, Porto Alegre, RS 91540-000, Brazil. E-mail: moleta@terra.com.br The leaves of Dodonea viscosa were force fed to five bovines. Four received the leaves in fresh green stage and the fifth dried ones. Clinical signs were observed in four of the bovines that died; the fifth did not show signs of poisoning. The fresh green plant was proved to be toxic from a dose of 25g/kg on. Dried leaves fed at the dose of 30 g/kg were also toxic. All the animals that died showed clinical signs from 13h30min to 45h after the ingestion of the plant and and death followed within about 48h. The clinical course lasted for about 8h30min until death. The main symptoms where apathy, anorexia, slight tenesmus, muscle trembling, difficulties to keep consciousness, pressing the head against obstacles, lateral recumbency, paddling movements, coma and death. The most significant macroscopic alterations were observed in the liver, with accentuation of the lobular pattern, dark-red areas interspersed with yellowish areas. Petechiae were found in serosal membranes of the abdominal and thoracic organs as well as the intestines. The main microscopic change was hepatic centrolobular coagulative necrosis, associated with congestion and hemorrhages.

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:

ABSTRACT.- Araújo ER., Madruga C.R., Soares e.o. & Kessler R.H. 2003. [Progresses in immunization against Anaplasma marginale] Progressos na imunização contra Anaplasma marginale. Pesquisa Veterinária Brasileira23(3):139-148. Embrapa Gado de Corte, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. E-mail: flabio@cnpgc.embrapa.br The current immunization against anaplasmosis in cattle is derived from the blood of infected animais, as live or dead organisms. Nevertheless, efforts have been made to develop a new generation of vaccines. Toe outer membrane of Anaplasma marginale induces a protective immune response against challenge with homologous isolates and a partially protective response against heterologous challenge. ln this membrane, six major surface proteins (MSPs) have been identified, which · have been targeted for the development of immunogens against anaplasmosis. From those proteins, MSP1 a and MSP2 have shown the greatest potential as immunogens, protecting cattle against challenge with virulent homologous and heterologous isolates of A. marginale, despite the size polymorphism of the former protein and the variability of the gene that encodes the latter protein. Another alternative of immunogen is the in vitro culture of A. marginale. lnactivated organisms originating from Dermacentor variabilis IDE8 cell culture were tested as immunogen. Cattle immunized with cell culture-derived A. marginale had a significantly lower reduction in the packed cell volume after challenge exposure and did not display clinical anaplasmosis. Besides the protection afforded by this type of immunogen, cell culture derived organisms are free from bovine cells and pathogens, what is a major advantage as compared with traditional immunization procedures.

• Anaplasma marginale Ehrlichiae immunization bovine major surface proteins culture imunização bovino proteínas principais de superfície cultura.

Abstract in Portuguese:

RESUMO.- Araújo ER., Madruga C.R., Soares e.o. & Kessler R.H. 2003. [Progresses in immunization against Anaplasma marginale] Progressos na imunização contra Anaplasma marginale. Pesquisa Veterinária Brasileira23(3):139-148. Embrapa Gado de Corte, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. E-mail: flabio@cnpgc.embrapa.br Até o presente momento, as imunizações contra anaplasmose em rebanhos bovinos utilizam organismos vivos ou mortos. No entanto, esforços têm sido realizados nos últimos anos com o objetivo de desenvolver uma nova geração de vacinas. A membrana externa de Anaplasma marginale é capaz de induzir reposta imune protetora contra desafio homólogo e parcialmente protetora contra desafio heterólogo. Nela foram identificadas seis proteínas principais de superfície (MSPs), as quais têm sido alvo de estudos para o desenvolvimento de imunógenos contra a anaplasmose. Destas proteínas, MSPla e MSP2 têm demonstrado maior potencial como imunógenos, protegendo os animais contra desafio com isolados virulentos homólogos e heterólogos de A margina/e, apesar do polimorfismo de tamanho da primeira proteína e variabilidade do gene que codifica a segunda. Uma outra alternativa para a imunização contra A. margina/e é o cultivo in vitro dessa riquétsia. Organismos inativados provenientes de cultivo em células IDE8 de Dermacentor variabilis foram testados como imunógeno. Os animais apresentaram uma significativa diferença na redução do volume globular após desafio e não apresentaram sinais clínicos de anaplasmose. Além da proteção conferida por este tipo de imunógeno, os organismos provenientes de cultura de células de carrapato são livres de células. e patógenos de bovinos, o que é uma vantagem significativa quando comparado aos processos tradicionais de imunização.

Abstract in English:

ABSTRACT.- Bonel-Raposoj., Driemeier D., Barros S.S. & Gevehr~Fernandes C. 2003. [Histological and ultrastructural evolution of liver lesions in experimental Myoporum laetum poisoning of sheep and cattle.] Evolução das lesões histológicas e ultra-estruturais no fígado de ovinos e bovinos experimentalmente intoxicados por Myoporum laetum. Pesquisa Veterinária Brasileira 23(4):149-155. Depto Patologia Animal, Faculdade de Veterinária, UFPel, Pelotas, RS 96010-900, Brazil. E-mail: bonel-raposo@brturbo.com Green leaves of Myoporum laetum were collected during spring and summer, and administered to five sheep and six steers at dosages of 20 and 30 g/kg. Liver biopsies were taken before (Controls) and 1, 3 and 7 days after dosage. ln sheep, the clinicai signs were depression, rumen hypomotility, dried feces, tenesmus, teeth grinding, dyspnea and typical lesions of photosensitization. ln cattle, the clinicai picture was much less pronounced. The main histological findings in sheep were vacuolization of hepatocytes, portal ftbrosis, bile duct proliferation and necrosis of periportal hepatocytes; the ultrastructural examination revealed hyperplasia of the smooth endoplasmic reticulum, hepatocellular hydropic degeneration, presence of crystals and severa! other degenerative changes. ln cattle both, the histological and the ultrastructural findings, were less evident.

• Poisonous plants plant poisoning Myoporum laetum sheep cattle Plantas tóxicas intoxicação por plantas ovinos bovinos.

Abstract in Portuguese:

RESUMO.- Bonel-Raposoj., Driemeier D., Barros S.S. & Gevehr~Fernandes C. 2003. [Histological and ultrastructural evolution of liver lesions in experimental Myoporum laetum poisoning of sheep and cattle.] Evolução das lesões histológicas e ultra-estruturais no fígado de ovinos e bovinos experimentalmente intoxicados por Myoporum laetum. Pesquisa Veterinária Brasileira 23(4):149-155. Depto Patologia Animal, Faculdade de Veterinária, UFPel, Pelotas, RS 96010-900, Brazil. E-mail: bonel-raposo@brturbo.com Amostras de Myoporum laetum foram colhidas durante a primavera e verão e administradas a cinco ovinos e seis bovinos em doses únicas de 20 e 30 g/kg. Biópsias hepáticas foram colhidas antes (controles) e 1, 3 e 7 dias após a dosagem da planta. Estas biópsias foram analisadas histológica e ultraestruturalmente. Os sinais clínicos, em ovinos, caracterizaramse, especialmente; pordepressão, diminuição dos movimentos ruminais, fezes ressequidas, tenesmo, ranger de dentes, dispnéia e lesões típicas de fotossensibilização. Em bovinos, o quadro clínico foi discreto. Os principais achados histológicos, em ovinos, incluíram vacuolização de hepatócitos, fibrose portal, proliferação de duetos biliares e necrose de hepatócitos periportais. Os estudos ultra-estruturais, em ovinos, revelaram hiperplasia do retículo endoplasmático liso, tumefação de hepatócitos, degranulação e vesiculação do retículo endoplasmático rugoso, presença de cristais aciculares, retenção biliar, tumefação de mitocôndrias e várias outras alterações degenerativas. Em bovinos, tanto os achados histológicos, quanto os ultra-estruturais foram menos evidentes.

Abstract in English:

ABSTRACT.- Azevedo S.S., Vasconcellos S.A., Alves C.J., Keid L.B., Grasso L.M.P.S., Mascolli R. & Pinheiro S.R. 2003. [Serological survey and risk factors for brucellosis due to Brucella canis in dogs of the Santana de Parnaíba municipality, State of São Paulo.] Inquérito sorológico e fatores de risco para a brucelose por Brucella canis em cães do Município de Santana de Parnaíba, Estado de São Paulo. Pesquisa Veterinária Brasileira 23(4):156-160. Depto Medicina Veterinária Preventiva, FMVZ/USP, 05508-090 São Paulo·SP, Brazil. E-mail: sevedo@fmvz.usp.br The prevalence of brucellosis due to Bruce/la canis was investigated in dogs of the Santana de Parnaíba. county, State of São Paulo, southeastem Brazil, and the risk factors for infection were analyzed. For this purpose, 41 O blood samples were collected from dogs duririg the rabies vaccination campaign, in August 1999. The agar gel immunodiffusion test (AGIO), using lipopolysaccharides and protein antigens from Brucella ovis; strain Reo 198, was applied first as a screening test on normal sera, and secondly, for confirmation. fl1e sarne AGIO test was applied to sera treated previously with 2-mercapthathanol (ME-AGIO). The complement fixation test (CFT), using B. ovis antigen, strain 63/290, was applied also as a confirmatory test. For the prevalence analysis, animais presenting positive results in both ME-AGIO and CFTwere considered positive. The prevalence of brucellosis due to B. canis was 2.2% (95% C.I. = 1.01-4.13%). Dogs that were allowed bytheir owners to stay free outside their home had a higher risk for contracting B. canis infection, with an odds ratio value of 8.73 (95% C.I.=1.48-51.55) and p=0.04.

• Animal brucellosis Brucella canis diagnosis dogs Brucelose animal diagnóstico cães.

Abstract in Portuguese:

RESUMO.- Azevedo S.S., Vasconcellos S.A., Alves C.J., Keid L.B., Grasso L.M.P.S., Mascolli R. & Pinheiro S.R. 2003. [Serological survey and risk factors for brucellosis due to Brucella canis in dogs of the Santana de Parnaíba municipality, State of São Paulo.] Inquérito sorológico e fatores de risco para a brucelose por Brucella canis em cães do Município de Santana de Parnaíba, Estado de São Paulo. Pesquisa Veterinária Brasileira 23(4):156-160. Depto Medicina Veterinária Preventiva, FMVZ/USP, 05508-090 São Paulo·SP, Brazil. E-mail: sevedo@fmvz.usp.br Foi investigada a prevalência da brucelose causada por Brucella canis em cães do município de Santana de Parnaíba, SP, Brasil, e realizado um estudo de possíveis fatores de risco associados à soropositividade para B. canis. Foram examinadas 410 amostras de soro sanguíneo de cães colhidas durante a campanha de vacinação anti-rábica animal, realizada em agosto de 1999. A imunodifusão em gel de ágar (IOGA), utilizando antígeno de lipopolissacarídeos e proteínas de Brucella ovis, amostra Reo 198, foi empregada em soros normais como teste de triagem, e, para a confirmação, a mesma técnica foi aplicada em soros tratados pelo 2-mercaptoetanol (IOGA-ME). A reação de fixação de complemento (CFT), utilizando antígeno de B. ovis, amostra 63/290, também foi utilizada como prova confirmatória. A determinação da prevalência considerou como positivos os animais que reagiram positivamente nos dois testes confirmatórios. (IOGA-ME e CFT). A prevalência da B. canis foi de 2,2% (1.C. 95% = 1,01-4, 13%). A análise estatística mostrou que os cães com acesso irrestrito à rua o dia todo (manejo do tipo solto) estiveram mais expostos ao risco da infecção por B. canis, com um valor de odds ratio de 8,73 (I.C. 95% = 1,48-51,55) e p=0,04.

Abstract in English:

ABSTRACT.- Fighera R.A., Rozza D.B., Piazer J.V., Copetti M.V., lrigoyen L.F. & Barros C.S.L. 2003. [Interstitial pneumonia in cattle fed moldy sweet potatoes (Ipomoea batatas).] Pneumonia intersticial em bovinos associada à ingestão de batata-doce (lpomoea batatas) mofada. Pesquisa Veterinária Brasileira 23(4):161-166. Depto Patologia, Universidade Federal de Santa Maria, 97105- 900 Santa Maria, RS, Brasil. E-mail: claudioslbarros@uol.com.br Cases of respiratory disease were diagnosed in five out of 23 cattle (21. 7%) after they wete fed moldy damaged sweet potatoes (lpomoea batatas) on a small farm in the county of São Vicente do Sul, state of Rio Grande do Sul, Brazil. Of those five cattle, three died spontaneously and another one was euthanatized for necropsy while showing advanced respiratory clinicai signs. The disease manifested itself approximately 24 hours after the ingestion of the sweet potatoes and lasted from 1 to 4 days. Clinicai signs included dyspnea (labored breathing and abdominal respiration), tachypnea, extended neck with Iow carriage of the head and rhythmical flaring of the nostrils. Two cows were necropsied. Necropsy findings included distended pale and rubbery lungs which failed to collapse when the thorax was open, and marked pulmonar interstitial emphysema and edema. Lymphoid hyperplasia was observed in the hilar nodes and spleen. Histologically, the lesions were those of interstitial pneumonia. Alveolar septa were thickened by fibroblasts and inflammatory cells, and there was hypertrophy and hyperplasia of type li pneumocytes; the interlobular septa were distended by edema and emphysema. The culture of the moldy sweet potatoes yielded Fusarium solani and Fusarium oxysporum.

• Toxic diseases mycotoxicoses respiratory diseases interstitial pneumonia diseases of cattle sweet potatoes lpomoea batatas Intoxicações micotoxicoses doenças respiratórias pneumonia intersticial doenças de bovinos batata-doce.

Abstract in Portuguese:

RESUMO.- Fighera R.A., Rozza D.B., Piazer J.V., Copetti M.V., lrigoyen L.F. & Barros C.S.L. 2003. [Interstitial pneumonia in cattle fed moldy sweet potatoes (Ipomoea batatas).] Pneumonia intersticial em bovinos associada à ingestão de batata-doce (lpomoea batatas) mofada. Pesquisa Veterinária Brasileira 23(4):161-166. Depto Patologia, Universidade Federal de Santa Maria, 97105- 900 Santa Maria, RS, Brasil. E-mail: claudioslbarros@uol.com.br Uma doença respiratória foi diagnosticada em cinco dentre 23 bovinos (21, 7%) após terem sido alimentados com batata-doce (lpomoea batatas) mofada em uma pequena propriedade rural em São Vicente do Sul, Rio Grande do Sul, Brasil. Três dos cinco bovinos afetados morreram espontaneamente, e o quarto foi sacrificado para necropsia quando mostrava sinais clínicos respiratórios avançados. A manifestação clínica iniciara cerca de 24 horas após a ingestão das batatasdoces e a evolução clínica foi de 1 a 4 dias. Os sinais clínicos incluíam dispnéia (respiração laboriosa e abdominal), taquipnéia, pescoço estendido com cabeça baixa e dilatação ritmada das narinas. Dois bovinos foram necropsiados. Os achados de necropsia incluíam pulmões distendidos, pálidos e de consistência borrachenta, que não colapsavam quando o tórax era aberto; enfisema e edema acentuados eram evidentes no pulmão. Os linfonodos e o baço apresentavam alterações características de hiperplasia linfóide. Histologicamente, as lesões eram típicas de pneumonia intersticial. Os septos alveolares estavam espessados por fibroblastos e células inflamatórias, havia hipertrofia e hiperplasia de pneumócitos tipo II; os septos interlobulares estavam distendidos por edema e enfisema. A cultura de amostras das batatas-doces mofadas produziu Fusarium solani e F. oxysporum.