PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

Amaral A.L., Morés N., Barioni W.J., Ventura L., Silva R.A.M. & Silva V.S. 2004. [Risk factors at the growth-termination phase associated with the occurrence of lymphadenitis in swine.] Fatores de risco associados à ocorrência de linfadenite em suínos na fase de crescimento-terminação. Pesquisa Veterinária Brasileira 24(3):120-122. Depto Epidemiologia, Embrapa Suínos e Aves, Cx. Postal 21, Concórdia, SC 89700-000, Brazil. E-mail: armando@cnpsa.embrapa.br An epidemiological study in 60 units of swine termination (UT), pertaining to producers integrated with the main Agribusiness of the States of Santa Catarina and Rio Grande do Sul. These units had been selected due to the occurrence of lymphadenitis in slaughtered pigs. The batch size studied in each UT varied from 22 to 960 pigs. The animals were considered positive or negative for lymphadenitis, following the evaluation routine of the Service of Federal Inspection (SIF) at slaughter. For data collection at the UTs, a questionnaire with 156 variables was applied at a single visit to the farms. In the 60 UT studied, the percentage of positive animals for lymphadenitis was 10.37%. Among the variables analyzed, nine risk factors were identified (explicative variables) and associated with the occurrence of lymphadenitis (objective variables). The explicative variables were: bad hygiene of the feeders and drinking fountains, untreated drinking water, bad conservation of the facilities, use of the same truck for ration and animal transportation, production of ration inside the farm, free access of other animals to the ingredients of ration, storage of ready usable ration in boxes or bags and handling of the production of animals in an continuous system.

• Swine risk factors lymphadenitis.

Abstract in Portuguese:

Amaral A.L., Morés N., Barioni W.J., Ventura L., Silva R.A.M. & Silva V.S. 2004. [Risk factors at the growth-termination phase associated with the occurrence of lymphadenitis in swine.] Fatores de risco associados à ocorrência de linfadenite em suínos na fase de crescimento-terminação. Pesquisa Veterinária Brasileira 24(3):120-122. Depto Epidemiologia, Embrapa Suínos e Aves, Cx. Postal 21, Concórdia, SC 89700-000, Brazil. E-mail: armando@cnpsa.embrapa.br An epidemiological study in 60 units of swine termination (UT), pertaining to producers integrated with the main Agribusiness of the States of Santa Catarina and Rio Grande do Sul. These units had been selected due to the occurrence of lymphadenitis in slaughtered pigs. The batch size studied in each UT varied from 22 to 960 pigs. The animals were considered positive or negative for lymphadenitis, following the evaluation routine of the Service of Federal Inspection (SIF) at slaughter. For data collection at the UTs, a questionnaire with 156 variables was applied at a single visit to the farms. In the 60 UT studied, the percentage of positive animals for lymphadenitis was 10.37%. Among the variables analyzed, nine risk factors were identified (explicative variables) and associated with the occurrence of lymphadenitis (objective variables). The explicative variables were: bad hygiene of the feeders and drinking fountains, untreated drinking water, bad conservation of the facilities, use of the same truck for ration and animal transportation, production of ration inside the farm, free access of other animals to the ingredients of ration, storage of ready usable ration in boxes or bags and handling of the production of animals in an continuous system.

Abstract in English:

Paulovich F.B., Borowski S.M., Driemeier D., Razia L.E., Coutinho T. A., Prates A.B.H., Pescador C., Correa A. & Barcellos D.E.S.N. 2004. [Assessment of the pathogenicity of Brachyspira pilosicoli strains by conventional histopathological techniques and by immunohistochemistry.] Avaliação da patogenicidade de amostras de Brachyspira pilosicoli através de técnicas histopatológicas convencionais e por imuno-histoquímica. Pesquisa Veterinária Brasileira 24(3):144-148. Depto Patologia Clínica Veterinária, Faculdade de Veterinária, UFRGS, Cx. Postal 15094, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@vortex.ufrgs.br The study assessed differences of pathogenicity among 19 strains of Brachyspira pilosicoli isolated from cases of diarrhea in swine in the State of Rio Grande do Sul, Brazil, using an one-day-old chick model. The chicken inoculated with live bacterial cultures were submitted to euthanasia 21 days later, and the cecum was examined histologically using hematoxylin-eosin, silver staining and immunohistochemistry with a polyclonal antibody anti-Brachyspira pilosicoli. With silver staining and immunohistochemistry, respectively, 21.59% and 70.96% of the chicken showed colonization of the cecal epithelium with B. pilosicoli. Differences in the type of colonization characterized by continuous adhesion, focal adhesion or free bacteria in the intestinal lumen were observed. Immunohistochemistry was more efficient to assess intestinal colonization, because 49.37% more colonized chicken could be detected with silver staining. With three strains, elongated figures inside the cytoplasm of epithelial cecal cells in the inoculated chicken were observed.

• Brachyspira spp Brachyspira pilosicoli spirochetal colitis immunohistochemistry pathogenesis.

Abstract in Portuguese:

Paulovich F.B., Borowski S.M., Driemeier D., Razia L.E., Coutinho T. A., Prates A.B.H., Pescador C., Correa A. & Barcellos D.E.S.N. 2004. [Assessment of the pathogenicity of Brachyspira pilosicoli strains by conventional histopathological techniques and by immunohistochemistry.] Avaliação da patogenicidade de amostras de Brachyspira pilosicoli através de técnicas histopatológicas convencionais e por imuno-histoquímica. Pesquisa Veterinária Brasileira 24(3):144-148. Depto Patologia Clínica Veterinária, Faculdade de Veterinária, UFRGS, Cx. Postal 15094, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@vortex.ufrgs.br The study assessed differences of pathogenicity among 19 strains of Brachyspira pilosicoli isolated from cases of diarrhea in swine in the State of Rio Grande do Sul, Brazil, using an one-day-old chick model. The chicken inoculated with live bacterial cultures were submitted to euthanasia 21 days later, and the cecum was examined histologically using hematoxylin-eosin, silver staining and immunohistochemistry with a polyclonal antibody anti-Brachyspira pilosicoli. With silver staining and immunohistochemistry, respectively, 21.59% and 70.96% of the chicken showed colonization of the cecal epithelium with B. pilosicoli. Differences in the type of colonization characterized by continuous adhesion, focal adhesion or free bacteria in the intestinal lumen were observed. Immunohistochemistry was more efficient to assess intestinal colonization, because 49.37% more colonized chicken could be detected with silver staining. With three strains, elongated figures inside the cytoplasm of epithelial cecal cells in the inoculated chicken were observed.

Abstract in English:

Rech R.R, Fighera R.A., Oliveira F.N. & Barros C.S.L. 2004. [Granulomatous meningoencephalitis in cattle grazing vetch (Vicia spp).] Meningoencefalite granulomatosa em bovinos em pastoreio de ervilhaca (Vicia spp). Pesquisa Veterinária Brasileira 24(3): 169-172. Depto Patologia, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. E-mail: claudioslbarros@uol.com.br Granulomatous meningoencephalitis was observed in 7 out of 8 adult dairy cows affected by vetch-associated systemic granulomatous disease, although there was no neurological signs associated with the condition. The cellular inflammatory infiltrates were located in the leptomeninges and as perivascular cuffings within the brain and consisted of epithelioid macrophages, lymphocytes, plasm cells and eosinophils. These inflammatory changes varied from mild to severe from animal to animal and among different brain regions of the same animal. Perivascular cuffings were usually more marked than the leptomeningeal infiltrates. Affected brain regions, in decreasing order of intensity, included diencephalon through the level of massa intermedia, mesencephalon at the level of rostral colliculi, pons and cerebellar peduncles, medulla at the level of obex, frontal lobe at the level of the genu of the corpus callosum, and cerebellum. The character and the distribution of the inflammatory changes are emphasized regarding the differential diagnosis with other diseases and lesions of the bovine central nervous system in the context of the Brazilian surveillance program for bovine spongiform encephalopath.

• Diseases of cattle diseases of the central nervous system meningoencephalitis plant poisoning Vicia spp pathology.

Abstract in Portuguese:

Rech R.R, Fighera R.A., Oliveira F.N. & Barros C.S.L. 2004. [Granulomatous meningoencephalitis in cattle grazing vetch (Vicia spp).] Meningoencefalite granulomatosa em bovinos em pastoreio de ervilhaca (Vicia spp). Pesquisa Veterinária Brasileira 24(3): 169-172. Depto Patologia, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. E-mail: claudioslbarros@uol.com.br Granulomatous meningoencephalitis was observed in 7 out of 8 adult dairy cows affected by vetch-associated systemic granulomatous disease, although there was no neurological signs associated with the condition. The cellular inflammatory infiltrates were located in the leptomeninges and as perivascular cuffings within the brain and consisted of epithelioid macrophages, lymphocytes, plasm cells and eosinophils. These inflammatory changes varied from mild to severe from animal to animal and among different brain regions of the same animal. Perivascular cuffings were usually more marked than the leptomeningeal infiltrates. Affected brain regions, in decreasing order of intensity, included diencephalon through the level of massa intermedia, mesencephalon at the level of rostral colliculi, pons and cerebellar peduncles, medulla at the level of obex, frontal lobe at the level of the genu of the corpus callosum, and cerebellum. The character and the distribution of the inflammatory changes are emphasized regarding the differential diagnosis with other diseases and lesions of the bovine central nervous system in the context of the Brazilian surveillance program for bovine spongiform encephalopath.

Abstract in English:

Lázaro N.S., Tibana A., Rodrigues D.P., Reis E.M.F., Quintaes B.R. & Hofer E. 2004. Antimicrobial resistance and R-plasmid in Salmonella spp from swine and abattoir environments. Pesquisa Veterinária Brasileira 24(2):57-60. Depto Epidemiologia e Saúde Pública, Instituto de Veterinária, UFRRJ, Seropédica, RJ 23890-000, Brazil. E-mail: nslazaro@aol.com Salmonella serovars isolated from swine are of particular interest not only because of the pathogenic potential for this animal species, but also due to its relevance with regard to public health. On basis of the profile of resistance to antimicrobials, 13 Salmonella strains were selected which belonged to the serovars Muenster (7), Derby (4), Typhimurium (1), and Braenderup (1). They were isolated from healthy swine as well as from the abattoir environment in the state of Rio de Janeiro. All strains of Salmonella were subjected to bacterial conjugation, and the E. coli K12 Nalr Lac+ F standard strain was used as receptor, with the purpose to verify the ability to transfer the resistance marks. Gene transfer phenomenon was detected in seven strains, and except Salmonella Typhimurium which transconjugated to Sm, Tc and Su, the remaining ones were characterized by transferring mark Su only. By plasmidial analysis of strains used and their respective transconjugants, 63 Kb plasmid was found, which was probably related to S. Typhimurium resistance.

• Salmonella swine antimicrobial resistance R-plasmid.

Abstract in Portuguese:

Lázaro N.S., Tibana A., Rodrigues D.P., Reis E.M.F., Quintaes B.R. & Hofer E. 2004. Antimicrobial resistance and R-plasmid in Salmonella spp from swine and abattoir environments. Pesquisa Veterinária Brasileira 24(2):57-60. Depto Epidemiologia e Saúde Pública, Instituto de Veterinária, UFRRJ, Seropédica, RJ 23890-000, Brazil. E-mail: nslazaro@aol.com Salmonella serovars isolated from swine are of particular interest not only because of the pathogenic potential for this animal species, but also due to its relevance with regard to public health. On basis of the profile of resistance to antimicrobials, 13 Salmonella strains were selected which belonged to the serovars Muenster (7), Derby (4), Typhimurium (1), and Braenderup (1). They were isolated from healthy swine as well as from the abattoir environment in the state of Rio de Janeiro. All strains of Salmonella were subjected to bacterial conjugation, and the E. coli K12 Nalr Lac+ F standard strain was used as receptor, with the purpose to verify the ability to transfer the resistance marks. Gene transfer phenomenon was detected in seven strains, and except Salmonella Typhimurium which transconjugated to Sm, Tc and Su, the remaining ones were characterized by transferring mark Su only. By plasmidial analysis of strains used and their respective transconjugants, 63 Kb plasmid was found, which was probably related to S. Typhimurium resistance.

Abstract in English:

Oliveira A., Fonseca A.H., Ishikawa M.M. & Yoshinari N.H. 2004. [Cinetic growth of Borrelia burgdorferi (Spirochaetaceae) in different culture media.] Cinética do crescimento de Borrelia burgdorferi em diferentes meios de cultivo. Pesquisa Veterinária Brasileira 24(2):61-64. Depto Epidemiologia e Saúde Pública, Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, Seropédica, RJ 23890-000, Brazil. E-mail: adivaldo@ufrrj.br The cinetic of growth of Borrelia burgdorferi was studied during a 3-month period, using the following 8 culture media: (1) rabbit serum BSK, (2) swine serum BSK, (3) swine serum BSK+5 fluorouracil, (4) PMR, (5) CTB, (6) Dubos, (7) Brucella broth and (8) BHI. All media were prepared aseptically and were maintained in culture tubes of 10 ml capacity. For each medium, the inoculum was standardized to contain initially 102 spirochetes for each 0.1 ml of culture. The growth was monitorized by counting the total number of spirochetes in 0.1ml of medium in a dark field microscope, using a 10x30 mm cover slip. For the first 12 days, counting was done each 24 hours, and afterwards once a week during 14 weeks. There occurred growth of B. burgdorferi in all tested media, with the best performance of three of them: BSK with rabbit serum, BSK swine serum + 5 fluorouracil, and CTB medium. Growth of B. burgdorferi was seen from the 4th week on, reaching its maximum within 8-12 weeks, depleting the culture medium after this time. Cystic forms of B. burgdorferi were observed with all tested media.

• Culture medium Borrelia burgdorferi borreliosis Spirochaetaceae bacteria cultivation cinetic growth cystic forms.

Abstract in Portuguese:

Oliveira A., Fonseca A.H., Ishikawa M.M. & Yoshinari N.H. 2004. [Cinetic growth of Borrelia burgdorferi (Spirochaetaceae) in different culture media.] Cinética do crescimento de Borrelia burgdorferi em diferentes meios de cultivo. Pesquisa Veterinária Brasileira 24(2):61-64. Depto Epidemiologia e Saúde Pública, Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, Seropédica, RJ 23890-000, Brazil. E-mail: adivaldo@ufrrj.br The cinetic of growth of Borrelia burgdorferi was studied during a 3-month period, using the following 8 culture media: (1) rabbit serum BSK, (2) swine serum BSK, (3) swine serum BSK+5 fluorouracil, (4) PMR, (5) CTB, (6) Dubos, (7) Brucella broth and (8) BHI. All media were prepared aseptically and were maintained in culture tubes of 10 ml capacity. For each medium, the inoculum was standardized to contain initially 102 spirochetes for each 0.1 ml of culture. The growth was monitorized by counting the total number of spirochetes in 0.1ml of medium in a dark field microscope, using a 10x30 mm cover slip. For the first 12 days, counting was done each 24 hours, and afterwards once a week during 14 weeks. There occurred growth of B. burgdorferi in all tested media, with the best performance of three of them: BSK with rabbit serum, BSK swine serum + 5 fluorouracil, and CTB medium. Growth of B. burgdorferi was seen from the 4th week on, reaching its maximum within 8-12 weeks, depleting the culture medium after this time. Cystic forms of B. burgdorferi were observed with all tested media.

Abstract in English:

Lázaro N.S., Tibana A., Reis E.M.F., Rodrigues D.P., Quintaes B.R. & Hofer E. 2004. [Antimicrobial susceptibility pattern and plasmid profile in Salmonella Muenster isolated from swine and abattoir environment, Brazil.] Padrão de susceptibilidade a antimicrobianos e perfil plasmidial em Salmonella Muenster isoladas de suínos e do ambiente de abatedouros. Pesquisa Veterinária Brasileira 24(2):65-70. Depto Epidemiologia e Saúde Pública, Inst.Veterinária, UFRRJ, Seropédica, RJ 23890-000, Brazil. E-mail: nslazaro@aol.com Thirty-eight strains of Salmonella Muenster, isolated from swine and the abattoir environment, in the State of Rio de Janeiro, Brazil, from March 1991 to February 1992, were analyzed for the presence of plasmids. The strains were selected according to their profile regarding the antimicrobials: streptomycin, tetracycline, sulphonamide and sulfametoxazole-trimethoprim. Thirteen strains were resistant to one or several antimicrobials, 18 with intermediate degree and seven were sensitive. Plasmids varying in size from 1.2 Kb to 42 Kb were detected in 37 (97.36%) of the 38 samples, corresponding to 11 different profiles (P1- P11), varying from 1 to 6 plasmids per model. The number and plasmids diversity was greater than the resistance marks for strains. The plasmid of 2.85 Kb was the most frequent, being present in 83.78% of the 37 strains; only the one of 7.5 Kb was detected at the two slaughterhouses. There was no parallelism between resistance pattern and plasmidial profile, and a same antibiotype was found in several plasmidial profiles. The results of the present investigation, allowed us to conclude that the plasmid characterization is an useful and simple tool for the epidemiological typing of this sorovar.

• Salmonella Muenster swine antimicrobial resistance plasmids.

Abstract in Portuguese:

Lázaro N.S., Tibana A., Reis E.M.F., Rodrigues D.P., Quintaes B.R. & Hofer E. 2004. [Antimicrobial susceptibility pattern and plasmid profile in Salmonella Muenster isolated from swine and abattoir environment, Brazil.] Padrão de susceptibilidade a antimicrobianos e perfil plasmidial em Salmonella Muenster isoladas de suínos e do ambiente de abatedouros. Pesquisa Veterinária Brasileira 24(2):65-70. Depto Epidemiologia e Saúde Pública, Inst.Veterinária, UFRRJ, Seropédica, RJ 23890-000, Brazil. E-mail: nslazaro@aol.com Thirty-eight strains of Salmonella Muenster, isolated from swine and the abattoir environment, in the State of Rio de Janeiro, Brazil, from March 1991 to February 1992, were analyzed for the presence of plasmids. The strains were selected according to their profile regarding the antimicrobials: streptomycin, tetracycline, sulphonamide and sulfametoxazole-trimethoprim. Thirteen strains were resistant to one or several antimicrobials, 18 with intermediate degree and seven were sensitive. Plasmids varying in size from 1.2 Kb to 42 Kb were detected in 37 (97.36%) of the 38 samples, corresponding to 11 different profiles (P1- P11), varying from 1 to 6 plasmids per model. The number and plasmids diversity was greater than the resistance marks for strains. The plasmid of 2.85 Kb was the most frequent, being present in 83.78% of the 37 strains; only the one of 7.5 Kb was detected at the two slaughterhouses. There was no parallelism between resistance pattern and plasmidial profile, and a same antibiotype was found in several plasmidial profiles. The results of the present investigation, allowed us to conclude that the plasmid characterization is an useful and simple tool for the epidemiological typing of this sorovar.

Abstract in English:

Beltrão N., Furian T.Q., Leão J.A., Pereira R.A., Moraes L.B. & Canal C.W. 2004. [Detection of infectious laryngotracheitis virus in chickens in Brazil.] Detecção do vírus da laringotraqueíte das galinhas no Brasil. Pesquisa Veterinária Brasileira 24(2):85-88. Centro de Diagnóstico e Pesquisa em Patologia Aviária (CDPA), Faculdade de Veterinária, UFRGS, Porto Alegre, RS 91540-000, Brazil. E-mail: nilzaneb@hotmail.com A study was carried out in search for evidences of infectious laryngotracheitis virus (ILTV) infections in some Brazilian chicken flocks. Tracheal tissues and swabs were collected from 10 different flocks of layers and broilers displaying respiratory signs of disease. Samples were processes for virus isolation in embryonated eggs and the membranes examined by histopathology. In addition, specimens were examined by polymerase chain reaction (PCR). Three flocks had ILTV positive chickens by virus isolation and PCR. These results confirm the occurrence of ILTV in chickens in Brazil.

• Infectious laryngotracheitis virus PCR virus isolation avian pathology.

Abstract in Portuguese:

Beltrão N., Furian T.Q., Leão J.A., Pereira R.A., Moraes L.B. & Canal C.W. 2004. [Detection of infectious laryngotracheitis virus in chickens in Brazil.] Detecção do vírus da laringotraqueíte das galinhas no Brasil. Pesquisa Veterinária Brasileira 24(2):85-88. Centro de Diagnóstico e Pesquisa em Patologia Aviária (CDPA), Faculdade de Veterinária, UFRGS, Porto Alegre, RS 91540-000, Brazil. E-mail: nilzaneb@hotmail.com A study was carried out in search for evidences of infectious laryngotracheitis virus (ILTV) infections in some Brazilian chicken flocks. Tracheal tissues and swabs were collected from 10 different flocks of layers and broilers displaying respiratory signs of disease. Samples were processes for virus isolation in embryonated eggs and the membranes examined by histopathology. In addition, specimens were examined by polymerase chain reaction (PCR). Three flocks had ILTV positive chickens by virus isolation and PCR. These results confirm the occurrence of ILTV in chickens in Brazil.

Abstract in English:

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:

Rossetti C.A., Vansco B.N., Pini, N & Carfagnini J.C. 2004. Comparison of three diagnostic techniques for the detection of leptospires in the kidneys of wild house mice (Mus musculus). Pesquisa Veterinária Brasileira 24(1):6-10. Instituto de Patobiología, Centro Nacional de Investigaciones Agropecuarias (CNIA) del Instituto Nacional de Tecnología Agropecuaria (INTA), CC 25 (1712) Castelar, Buenos Aires, Argentina. E-mail: crossetti@cicv.inta.gov.ar Forty-one wild house mice (Mus musculus) were trapped in an urban area, near railways, in Santa Fe city, Argentina. Both kidneys from each mouse were removed for bacteriological and histological examination. One kidney was inoculated into Fletcher semi-solid medium and isolates were serologically typed. The other kidney was microscopically examined after hematoxylin-eosin, silver impregnation and immunohistochemical stains. Leptospires, all of them belonging to the Ballum serogroup, were isolated from 16 (39%) out of 41 samples. The presence of the agent was recorded in 18 (44%) and in 19 (46%) out of 41 silver impregnated and immunohistochemically stained samples respectively. Additionally, leptospires were detected in high number on the apical surface of epithelial cells and in the lumen of medullary tubules and they were less frequently seen on the apical surface of epithelial cells or in the lumen of the cortical tubules, which represents an unusual finding in carrier animals. Microscopic lesions consisting of focal mononuclear interstitial nephritis, glomerular shrinkage and desquamation of tubular epithelial cells were observed in 13 of 19 infected and in 10 of 22 non-infected mice; differences in presence of lesions between infected and non-infected animals were not statistically significant (P=0,14). The three techniques, culture, silver impregnation and immunohistochemistry, had a high agreement (k³0.85) and no significant differences between them were detected (P>0.05). In addition, an unusual location of leptospires in kidneys of carrier animals was reported, but a relationship between lesions and presence of leptospires could not be established.

• Leptospires diagnostic techniques Mus musculus lesions.

Abstract in Portuguese:

Rossetti C.A., Vansco B.N., Pini, N & Carfagnini J.C. 2004. Comparison of three diagnostic techniques for the detection of leptospires in the kidneys of wild house mice (Mus musculus). Pesquisa Veterinária Brasileira 24(1):6-10. Instituto de Patobiología, Centro Nacional de Investigaciones Agropecuarias (CNIA) del Instituto Nacional de Tecnología Agropecuaria (INTA), CC 25 (1712) Castelar, Buenos Aires, Argentina. E-mail: crossetti@cicv.inta.gov.ar Forty-one wild house mice (Mus musculus) were trapped in an urban area, near railways, in Santa Fe city, Argentina. Both kidneys from each mouse were removed for bacteriological and histological examination. One kidney was inoculated into Fletcher semi-solid medium and isolates were serologically typed. The other kidney was microscopically examined after hematoxylin-eosin, silver impregnation and immunohistochemical stains. Leptospires, all of them belonging to the Ballum serogroup, were isolated from 16 (39%) out of 41 samples. The presence of the agent was recorded in 18 (44%) and in 19 (46%) out of 41 silver impregnated and immunohistochemically stained samples respectively. Additionally, leptospires were detected in high number on the apical surface of epithelial cells and in the lumen of medullary tubules and they were less frequently seen on the apical surface of epithelial cells or in the lumen of the cortical tubules, which represents an unusual finding in carrier animals. Microscopic lesions consisting of focal mononuclear interstitial nephritis, glomerular shrinkage and desquamation of tubular epithelial cells were observed in 13 of 19 infected and in 10 of 22 non-infected mice; differences in presence of lesions between infected and non-infected animals were not statistically significant (P=0,14). The three techniques, culture, silver impregnation and immunohistochemistry, had a high agreement (k³0.85) and no significant differences between them were detected (P>0.05). In addition, an unusual location of leptospires in kidneys of carrier animals was reported, but a relationship between lesions and presence of leptospires could not be established.

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.