PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Barbosa R.C., Riet-Correa F., Lima E.F., Medeiros R.M.T., Guedes K.M.R, Gardner D.R., Molyneux R.J. & Melo L.E.H. 2007. Experimental swainsonine poisoning in goats ingesting Ipomoea sericophylla and Ipomoea riedelii (Convolvulaceae). Pesquisa Veterinária Brasileira 27(10):409-414. Hospital Veterinário, CSTR, Universidade Federal de Campina Grande. Campus de Patos, 58700-000 Patos, Paraíba, Brazil. E-mail: franklin.riet@pq.cnpq.br Ipomoea sericophylla and Ipomoea riedelii cause a glycoprotein storage disease in goats. This paper reports the experimental poisoning in goats by dried I. sericophylla and I. riedelii containing 0.05% and 0.01% swainsonine, respectively. Three groups with four animals each were used. Group 1 received daily doses of 2g/kg body weight (bw) of dried I. sericophylla (150mg of swainsonine/kg). Goats from this group had clinical signs 36-38 days after the start of ingestion. Group 2 received dried I. riedelii daily doses of 2g/kg of I. riedelii (30mg of swainsonine/kg) for 70 days. No clinical signs were observed, therefore the swainsonine dose was increased to 60mg/kg for another 70 days. Goats from Group 2 had clinical signs 26-65 days after increase in swainsonine dose to 60mg/kg. Group 3 was used as control. In these experiments the minimum toxic dose was 60mg/kg which represents 0.0004% of the dry matter in goats ingesting 1.5% bw of the dry matter. For goats ingesting 2%-2.5% bw of dry matter this dose would be 0.00024%-0.0003% of the dry matter. After the end of the experiment two goats were euthanized and another six were observed for recovery of clinical signs. Four goats that continued to consume swainsonine containing plant for 39-89 days after the first clinical signs had non reversible signs, while two goats that ingested the plant for only 15 and 20 days after the first clinical signs recovered completely. These and previous results indicate that irreversible lesions due to neuronal loss occur in goats that continue to ingest the plants for about 30 days after the first clinical signs. Clinical signs and histological lesions were similar to those reported previously for goats poisoned by swainsonine containing plants. No significant alterations were found in packed cell volume, red and white blood cell counts, hemoglobin and mean corpuscular hemoglobin concentrations, mean corpuscular volume, and serum levels of glucose, total protein, and albumin, and the serum activities of gamma glutamyl transferase and aspartate aminotransferase. Swainsonine concentration of 0.05% in I. sericophylla and 0.01% in I. riedelii are different from samples of these plants used in previous experiments, which contained 0.14% and 0.5% swainsonine, respectively, demonstrating a wide variation in the toxicity of different samples.

• Poisonous plants plant poisoning lysosomal storage disease swainsonine goats Ipomoea sericophylla Ipomoea riedelii

Abstract in Portuguese:

ABSTRACT.- Barbosa R.C., Riet-Correa F., Lima E.F., Medeiros R.M.T., Guedes K.M.R, Gardner D.R., Molyneux R.J. & Melo L.E.H. 2007. Experimental swainsonine poisoning in goats ingesting Ipomoea sericophylla and Ipomoea riedelii (Convolvulaceae). Pesquisa Veterinária Brasileira 27(10):409-414. Hospital Veterinário, CSTR, Universidade Federal de Campina Grande. Campus de Patos, 58700-000 Patos, Paraíba, Brazil. E-mail: franklin.riet@pq.cnpq.br Ipomoea sericophylla and Ipomoea riedelii cause a glycoprotein storage disease in goats. This paper reports the experimental poisoning in goats by dried I. sericophylla and I. riedelii containing 0.05% and 0.01% swainsonine, respectively. Three groups with four animals each were used. Group 1 received daily doses of 2g/kg body weight (bw) of dried I. sericophylla (150mg of swainsonine/kg). Goats from this group had clinical signs 36-38 days after the start of ingestion. Group 2 received dried I. riedelii daily doses of 2g/kg of I. riedelii (30mg of swainsonine/kg) for 70 days. No clinical signs were observed, therefore the swainsonine dose was increased to 60mg/kg for another 70 days. Goats from Group 2 had clinical signs 26-65 days after increase in swainsonine dose to 60mg/kg. Group 3 was used as control. In these experiments the minimum toxic dose was 60mg/kg which represents 0.0004% of the dry matter in goats ingesting 1.5% bw of the dry matter. For goats ingesting 2%-2.5% bw of dry matter this dose would be 0.00024%-0.0003% of the dry matter. After the end of the experiment two goats were euthanized and another six were observed for recovery of clinical signs. Four goats that continued to consume swainsonine containing plant for 39-89 days after the first clinical signs had non reversible signs, while two goats that ingested the plant for only 15 and 20 days after the first clinical signs recovered completely. These and previous results indicate that irreversible lesions due to neuronal loss occur in goats that continue to ingest the plants for about 30 days after the first clinical signs. Clinical signs and histological lesions were similar to those reported previously for goats poisoned by swainsonine containing plants. No significant alterations were found in packed cell volume, red and white blood cell counts, hemoglobin and mean corpuscular hemoglobin concentrations, mean corpuscular volume, and serum levels of glucose, total protein, and albumin, and the serum activities of gamma glutamyl transferase and aspartate aminotransferase. Swainsonine concentration of 0.05% in I. sericophylla and 0.01% in I. riedelii are different from samples of these plants used in previous experiments, which contained 0.14% and 0.5% swainsonine, respectively, demonstrating a wide variation in the toxicity of different samples.

Abstract in English:

Abstract.- Brocchi M., Ferreira A., Lancellotti M., Stehling E.G., Campos T.A., Nakazato G., Pestana de Castro A.F. & Silveira W.D. 2006. Typing of avian pathogenic Escherichia coli strains by REP-PCR. Pesquisa Veterinária Brasileira 26(2):69-73. Departamento de Microbiologia e Imunologia, Instituto de Biologia, Universidade de Campinas, Cx. Postal 6109, Campinas, SP 13081-862, Brazil. E-mail: wds@unicamp.br In the present study the repetitive extragenic palindromic (REP) polymerase chain reaction (PCR) technique was used to establish the clonal variability of 49 avian Escherichia coli (APEC) strains isolated from different outbreak cases of septicemia (n=24), swollen head syndrome (n=14) and omphalitis (n=11). Thirty commensal strains isolated from poultry with no signs of these illnesses were used as control strains. The purified DNA of these strains produced electrophoretic profiles ranging from 0 to 15 bands with molecular sizes varying from 100 bp to 6.1 kb, allowing the grouping of the 79 strains into a dendrogram containing 49 REP-types. Although REP-PCR showed good discriminating power it was not able to group the strains either into specific pathogenic classes or to differentiate between pathogenic and non-pathogenic strains. On the contrary, we recently demonstrated that other techniques such as ERIC-PCR and isoenzyme profiles are appropriate to discriminate between commensal and APEC strains and also to group these strains into specific pathogenic classes. In conclusion, REP-PCR seems to be a technique neither efficient nor universal for APEC strains discrimination. However, the population clonal structure obtained with the use of REP-PCR must not be ignored particularly if one takes into account that the APEC pathogenic mechanisms are not completely understood yet.

• Avian Escherichia coli typing REP-PCR.

Abstract in Portuguese:

Abstract.- Brocchi M., Ferreira A., Lancellotti M., Stehling E.G., Campos T.A., Nakazato G., Pestana de Castro A.F. & Silveira W.D. 2006. Typing of avian pathogenic Escherichia coli strains by REP-PCR. Pesquisa Veterinária Brasileira 26(2):69-73. Departamento de Microbiologia e Imunologia, Instituto de Biologia, Universidade de Campinas, Cx. Postal 6109, Campinas, SP 13081-862, Brazil. E-mail: wds@unicamp.br In the present study the repetitive extragenic palindromic (REP) polymerase chain reaction (PCR) technique was used to establish the clonal variability of 49 avian Escherichia coli (APEC) strains isolated from different outbreak cases of septicemia (n=24), swollen head syndrome (n=14) and omphalitis (n=11). Thirty commensal strains isolated from poultry with no signs of these illnesses were used as control strains. The purified DNA of these strains produced electrophoretic profiles ranging from 0 to 15 bands with molecular sizes varying from 100 bp to 6.1 kb, allowing the grouping of the 79 strains into a dendrogram containing 49 REP-types. Although REP-PCR showed good discriminating power it was not able to group the strains either into specific pathogenic classes or to differentiate between pathogenic and non-pathogenic strains. On the contrary, we recently demonstrated that other techniques such as ERIC-PCR and isoenzyme profiles are appropriate to discriminate between commensal and APEC strains and also to group these strains into specific pathogenic classes. In conclusion, REP-PCR seems to be a technique neither efficient nor universal for APEC strains discrimination. However, the population clonal structure obtained with the use of REP-PCR must not be ignored particularly if one takes into account that the APEC pathogenic mechanisms are not completely understood yet.

Abstract in English:

Veschi J.L.A., Dutra I.S., Miyakawa M.E.F., Perri S.H.V. & Uzal F.A. 2006. Immunophrophylactic strategies against enterotoxemia caused by Clostridium perfringens type D in goats. Pesquisa Veterinária Brasileira 26(1):51-54. Departamento de Produção e Saúde Animal, Universidade Estadual Paulista, Rua Clóvis Pestana 793, Araçatuba, SP 16050-680, Brazil. E-mail: isdutra@fmva.unesp.br The serological response to an experimental vaccine against Clostridium perfringens type D enterotoxemia was evaluated in four groups of goats. Group 1 received colostrum from unvaccinated cows and no vaccine. Groups 2, 3 and 4 received colostrum from vaccinated cows. In addition, Groups 3 and 4 received a vaccine dose at 80 days of age, and Group 4 received a second vaccine dose at 120 days of age. Serum antibody levels were determined by ELISA in cows before and after calving, and in goats at 3, 80, 120 and 160 days of age. No significant difference in serum antibody levels was observed between vaccinated and unvaccinated cows, or between the four groups of goats evaluated at 3 days of life. Groups 3 and 4 presented mean antibody titers of 0.6 and 1.1 IU/ml, respectively, 40 days after first vaccination. The vaccine response of Group 4 was 1.8 IU/ml 40 days after the booster dose and was higher than that observed for Group 3 (0.2 IU/ml). Thus, in the proposed regimen the use of heterologous colostrum did not induce passive immunization in goat kids. However, first vaccination and a booster dose after 40 days triggered satisfactory antibody levels.

• Clostridium perfringens type D goat enterotoxemia immunoprophylactic strategies colostrum vaccine.

Abstract in Portuguese:

Veschi J.L.A., Dutra I.S., Miyakawa M.E.F., Perri S.H.V. & Uzal F.A. 2006. Immunophrophylactic strategies against enterotoxemia caused by Clostridium perfringens type D in goats. Pesquisa Veterinária Brasileira 26(1):51-54. Departamento de Produção e Saúde Animal, Universidade Estadual Paulista, Rua Clóvis Pestana 793, Araçatuba, SP 16050-680, Brazil. E-mail: isdutra@fmva.unesp.br The serological response to an experimental vaccine against Clostridium perfringens type D enterotoxemia was evaluated in four groups of goats. Group 1 received colostrum from unvaccinated cows and no vaccine. Groups 2, 3 and 4 received colostrum from vaccinated cows. In addition, Groups 3 and 4 received a vaccine dose at 80 days of age, and Group 4 received a second vaccine dose at 120 days of age. Serum antibody levels were determined by ELISA in cows before and after calving, and in goats at 3, 80, 120 and 160 days of age. No significant difference in serum antibody levels was observed between vaccinated and unvaccinated cows, or between the four groups of goats evaluated at 3 days of life. Groups 3 and 4 presented mean antibody titers of 0.6 and 1.1 IU/ml, respectively, 40 days after first vaccination. The vaccine response of Group 4 was 1.8 IU/ml 40 days after the booster dose and was higher than that observed for Group 3 (0.2 IU/ml). Thus, in the proposed regimen the use of heterologous colostrum did not induce passive immunization in goat kids. However, first vaccination and a booster dose after 40 days triggered satisfactory antibody levels.

Abstract in English:

Penatti M.P.A., Silva A.S., Valadares G.F. & Leite D.S. 2005. Occurrence of F42 colonization factor in Escherichia coli strains isolated from piglets with diarrhea. Pesquisa Veterinária Brasileira 25(1):31-33. Depto Microbiologia e Imunologia, Instituto de Biologia, Unicamp, Campinas, SP 13081-970, Brazil. E-mail: domingos@unicamp.br The objective of this study was to determine the presence of the colonization factor F42 in 168 strains of Escherichia coli isolated from diarrheic stools of newborn piglets. The presence of F42 in 12 (7.1%) strains was detected with the agglutination test. Through the Polymerase Chain Reaction (PCR) of F42 positive strains, gene encoding enterotoxins (ST-I, ST-II, LT-I and LT-II) were detected. The finding of ST-I/ST-II genes in 50% of the strains, ST-I (16%) and ST-II (25%) indicates a strong association of FC F42 with heat-stable enterotoxins (91%). In contrast, the thermolabile enterotoxin (LT-I and LT-II) genes were not detected. Serogroups of F42 positive strains were determined, serogroup O8 being the most prevalent (41,7%). Other serogroups, as there are O9, O11, O18, O32, O35, O98 and O101, were also identified. Thus, FC F42 was confirmed as an additional factor of virulence in the pathogenesis of porcine colibacillosis.

• Escherichia coli F42 piglets fimbriae diarrhea PCR.

Abstract in Portuguese:

Penatti M.P.A., Silva A.S., Valadares G.F. & Leite D.S. 2005. Occurrence of F42 colonization factor in Escherichia coli strains isolated from piglets with diarrhea. Pesquisa Veterinária Brasileira 25(1):31-33. Depto Microbiologia e Imunologia, Instituto de Biologia, Unicamp, Campinas, SP 13081-970, Brazil. E-mail: domingos@unicamp.br The objective of this study was to determine the presence of the colonization factor F42 in 168 strains of Escherichia coli isolated from diarrheic stools of newborn piglets. The presence of F42 in 12 (7.1%) strains was detected with the agglutination test. Through the Polymerase Chain Reaction (PCR) of F42 positive strains, gene encoding enterotoxins (ST-I, ST-II, LT-I and LT-II) were detected. The finding of ST-I/ST-II genes in 50% of the strains, ST-I (16%) and ST-II (25%) indicates a strong association of FC F42 with heat-stable enterotoxins (91%). In contrast, the thermolabile enterotoxin (LT-I and LT-II) genes were not detected. Serogroups of F42 positive strains were determined, serogroup O8 being the most prevalent (41,7%). Other serogroups, as there are O9, O11, O18, O32, O35, O98 and O101, were also identified. Thus, FC F42 was confirmed as an additional factor of virulence in the pathogenesis of porcine colibacillosis.

Abstract in English:

Seitz A.L., Colodel E.M., Barros S.S. & Driemeier D. 2005. [Experimental poisoning by Sida carpinifolia (Malvaceae) in sheep.] Intoxicação experimental por Sida carpinifolia (Malvaceae) em ovinos. Pesquisa Veterinária Brasileira 25(1):15-20. Departamento de Patologia Clínica Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS 91540-000, Brazil. E-mail: alseitz@terra.com.br. Seven sheep received dry crushed Sida carpinifolia L.f. One of them died at 18 and other at 53 days of the experiment. Four others were euthanatized and necropsied at 30, 45, 75 and 100 days. For one sheep the supply of S. carpinifolia was interrupted on the 80th day of the experiment, and 70 days later the animal was euthanized and necropsied. The minimal amount of the dry plant consumed was 11 g/kg and the maximum was 30 g/kg. The progression of clinical findings was similar in six animals with slight diarrhea at 20 days of experiment. Neurological signs were observed at 25 days and included ataxia with dysmetria, muscle tremors of the head, atypical postural reactions, frequent falls, sluggish of movements, difficulty in grazing and swallowing. These signs were enhanced when the animals were forced to walk. Four of the animals presented progressive emaciation. The sheep whose supply of the plant was interrupted recovered gradually, and 11 days after the animal returned to normal. During necropsy, only enlarged mesenteric lymph nodes were observed. The histological alterations were more significant in the central nervous system, with multiple and severe cytoplasmic distention and vacuolation which affects specially Purkinje cells of the cerebellum, neurons of cerebral cortex, thalamus, midbrain and the ventral horn of spinal cord. Axonal spheroids in the brain, more frequently in the granular layer of cerebellum were also observed. The cytoplasmic vacuolation was also found in pancreatic acinar cells, renal tubules, thyroid follicular epithelium, hepatocytes and macrophages of lymphoid organs. The ultrastructural lesions observed were cytoplasmic vacuolation, some surrounded by membranes in Purkinje cells of cerebellum and thyroid follicular cells. The sheep, which had S. carpinifolia withdrawn from its diet for 70 days, had no significant histological alterations.

• Toxic plants plant poisoning Sida carpinifolia Malvaceae lysosomal storage disease swainsonine electronic microscopy.

Abstract in Portuguese:

Seitz A.L., Colodel E.M., Barros S.S. & Driemeier D. 2005. [Experimental poisoning by Sida carpinifolia (Malvaceae) in sheep.] Intoxicação experimental por Sida carpinifolia (Malvaceae) em ovinos. Pesquisa Veterinária Brasileira 25(1):15-20. Departamento de Patologia Clínica Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS 91540-000, Brazil. E-mail: alseitz@terra.com.br. Seven sheep received dry crushed Sida carpinifolia L.f. One of them died at 18 and other at 53 days of the experiment. Four others were euthanatized and necropsied at 30, 45, 75 and 100 days. For one sheep the supply of S. carpinifolia was interrupted on the 80th day of the experiment, and 70 days later the animal was euthanized and necropsied. The minimal amount of the dry plant consumed was 11 g/kg and the maximum was 30 g/kg. The progression of clinical findings was similar in six animals with slight diarrhea at 20 days of experiment. Neurological signs were observed at 25 days and included ataxia with dysmetria, muscle tremors of the head, atypical postural reactions, frequent falls, sluggish of movements, difficulty in grazing and swallowing. These signs were enhanced when the animals were forced to walk. Four of the animals presented progressive emaciation. The sheep whose supply of the plant was interrupted recovered gradually, and 11 days after the animal returned to normal. During necropsy, only enlarged mesenteric lymph nodes were observed. The histological alterations were more significant in the central nervous system, with multiple and severe cytoplasmic distention and vacuolation which affects specially Purkinje cells of the cerebellum, neurons of cerebral cortex, thalamus, midbrain and the ventral horn of spinal cord. Axonal spheroids in the brain, more frequently in the granular layer of cerebellum were also observed. The cytoplasmic vacuolation was also found in pancreatic acinar cells, renal tubules, thyroid follicular epithelium, hepatocytes and macrophages of lymphoid organs. The ultrastructural lesions observed were cytoplasmic vacuolation, some surrounded by membranes in Purkinje cells of cerebellum and thyroid follicular cells. The sheep, which had S. carpinifolia withdrawn from its diet for 70 days, had no significant histological alterations.

Abstract in English:

Minho A.P., Navarro I.T., Freire R.L., Vidotto O., Gennari S.M., Marana E.M. & Garcia J.L. 2004. Evaluation of the indirect fluorescent antibody test and modified agglutination test for detection of antibodies against Toxoplasma gondii in experimentally infected pigs. Pesquisa Veterinária Brasileira 24(4):199-202. Depto Medicina Veterinária Preventiva, Universidade Estadual de Londrina, Cx. Postal 6001, Londrina, PR 86050-970, Brazil. E-mail: italmar@uel.br The study determined the sensitivity and specificity of the indirect fluorescent antibody test (IFAT) and modified agglutination test (MAT) for anti-Toxoplasma gondii antibody detection by analyzing sera from 46 experimentally infected pigs. Values for sensitivity were 95.7% (confidence interval 95%: 84.0-99.2%) and for specificity 97.8% (confidence interval 95%: 87.0-99.9%) in both tests. There was an optimum agreement of results between IFAT and MAT evidenced by a Kappa test of 0.86. These results validate these tests for the detection of T. gondii infection in pigs. IFAT and MAT despite methodologies with different characteristics and readings have similar accuracy in pig serum samples.

• Toxoplasma gondii swine antibodies serology indirect fluorescent antibody test (IFAT) modified agglutination test (MAT).

Abstract in Portuguese:

Minho A.P., Navarro I.T., Freire R.L., Vidotto O., Gennari S.M., Marana E.M. & Garcia J.L. 2004. Evaluation of the indirect fluorescent antibody test and modified agglutination test for detection of antibodies against Toxoplasma gondii in experimentally infected pigs. Pesquisa Veterinária Brasileira 24(4):199-202. Depto Medicina Veterinária Preventiva, Universidade Estadual de Londrina, Cx. Postal 6001, Londrina, PR 86050-970, Brazil. E-mail: italmar@uel.br The study determined the sensitivity and specificity of the indirect fluorescent antibody test (IFAT) and modified agglutination test (MAT) for anti-Toxoplasma gondii antibody detection by analyzing sera from 46 experimentally infected pigs. Values for sensitivity were 95.7% (confidence interval 95%: 84.0-99.2%) and for specificity 97.8% (confidence interval 95%: 87.0-99.9%) in both tests. There was an optimum agreement of results between IFAT and MAT evidenced by a Kappa test of 0.86. These results validate these tests for the detection of T. gondii infection in pigs. IFAT and MAT despite methodologies with different characteristics and readings have similar accuracy in pig serum samples.

Abstract in English:

Canal C.W., Ferreira D.J., Macagnan M., Fallavena L.C.B., Moraes H.L.S. & Wald V.B. 2004. Prevalence of antibodies against chicken anaemia virus (CAV) in broiler breeders in Southern Brazil. Pesquisa Veterinária Brasileira 24(2):89-92. Centro de Diagnóstico e Pesquisa em Patologia Aviária (CDPA), Faculdade de Veterinária da Universidade Federal do Rio Grande do Sul, Porto Alegre, RS 91540-000, Brazil. E-mail: claudio.canal@ufrgs.br Chicks infected during the first two weeks of life with chicken anaemia virus (CAV) manifest clinical disease that can be avoided if the breeder hens transfer enough antibodies to their progeny. The objective of the present work was to establish the prevalence and titer of anti-CAV antibodies in some Brazilian broiler hen breeder flocks and verify in which phase of life the birds were infected. A total of 1,709 serum samples from 12 broiler hen flocks vaccinated against CAV and 64 unvaccinated flocks were analyzed for CAV antibodies with an enzyme-linked immunosorbent assay (ELISA). All non-vaccinated breeder flocks were found to be infected with CAV, with 89% of the hens tested presenting antibodies, 52% of these with titers considered high enough to protect their progeny against CAV infection. Likewise, all vaccinated hens had antibody titer to CAV capable of conferring protection to their progeny. Thus, vaccination of hens seems capable of conferring protection to chicks against clinically apparent CAV-associated disease.

• Antibodies avian pathology chicken anemia virus CAV epidemiology prevalence.

Abstract in Portuguese:

Canal C.W., Ferreira D.J., Macagnan M., Fallavena L.C.B., Moraes H.L.S. & Wald V.B. 2004. Prevalence of antibodies against chicken anaemia virus (CAV) in broiler breeders in Southern Brazil. Pesquisa Veterinária Brasileira 24(2):89-92. Centro de Diagnóstico e Pesquisa em Patologia Aviária (CDPA), Faculdade de Veterinária da Universidade Federal do Rio Grande do Sul, Porto Alegre, RS 91540-000, Brazil. E-mail: claudio.canal@ufrgs.br Chicks infected during the first two weeks of life with chicken anaemia virus (CAV) manifest clinical disease that can be avoided if the breeder hens transfer enough antibodies to their progeny. The objective of the present work was to establish the prevalence and titer of anti-CAV antibodies in some Brazilian broiler hen breeder flocks and verify in which phase of life the birds were infected. A total of 1,709 serum samples from 12 broiler hen flocks vaccinated against CAV and 64 unvaccinated flocks were analyzed for CAV antibodies with an enzyme-linked immunosorbent assay (ELISA). All non-vaccinated breeder flocks were found to be infected with CAV, with 89% of the hens tested presenting antibodies, 52% of these with titers considered high enough to protect their progeny against CAV infection. Likewise, all vaccinated hens had antibody titer to CAV capable of conferring protection to their progeny. Thus, vaccination of hens seems capable of conferring protection to chicks against clinically apparent CAV-associated disease.

Abstract in English:

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.- Vianni M.C.E. & Lázaro N.S. 2003. [Profile of antimicrobial susceptibility in strains of Gram positive cocci, negative catalase, isolated from buffalo subclinical mastitis.] Perfil de susceptibilidade a antimicrobianos em amostras de cocos Gram-positivos, catalase negativos, isoladas de mastite subclínica bubalina. Pesquisa Veterinária Brasileira 23(2):47-51. Depto Epidemiologia e Saúde Pública, Instituto de Veterinária, UFRRJ, Seropédica, RJ 23890-000, Brazil. The susceptibility of antimicrobials was studied in Gram positive and catalase negative cocci (21 samples of Lactococcus garvieae and 6 Enterococcus gallinarum), isolated from the milk of cows with subclinical mastitis, belonging to six buffalo herds in the State of Rio de Janeiro. The test used was diffusion of disks in agar Müller Hinton, according to recommendations of the National Committee for Clinicai Laboratory Standards - NCCLS. There were tested disks with ampicillin (10mg), cefalotin (30mg), cefotaxime (30mg), cefoxitin (30mg), doranfenicol (30mg), eritromycin (15mg), gentamycin (10mg), nitrofurantoin (300mg), norfloxacin (10mg), penicillin (1 O IU), tetracydin (30mg) and vancomycin (30mg). The results showed that with Lactococcus garvieae, the most efficient antimicrobial was nitrofurantoin, revealing 85.71% sensibility, followed by cefotaxime (61.90%), vancomycin (52.38%), norfloxacin (47.62%) and cefalotin (47.62%). The highest resistance was developed against penicillin and ampicillin, with 95.24% resistance for the two antimicrobials. The susceptibility profile developed by the strains of Enterococcus gallinarum showed low sensibility against the tested antimicrobials; the highest resistance observed was against eritromycin and gentamycin, with 33.34% sensibility for both. The antimicrobial evaluation showed 100% resistance against vancomycin and tetracyclin, followed by cloranfenicol, penicillin, ampicillin, cefoxitin, cefotaxim, norfloxacin and nitrofurantoin; all of them showed a resistance of 83.33% with the samples tested.

• Buffalo subclinical mastitis antimicrobialsusceptibility Gram positive cocci Mastite subclínica bubalina susceptibilidade a antimicrobianos cocos Gram-positivos.

Abstract in Portuguese:

RESUMO.- Vianni M.C.E. & Lázaro N.S. 2003. [Profile of antimicrobial susceptibility in strains of Gram positive cocci, negative catalase, isolated from buffalo subclinical mastitis.] Perfil de susceptibilidade a antimicrobianos em amostras de cocos Gram-positivos, catalase negativos, isoladas de mastite subclínica bubalina. Pesquisa Veterinária Brasileira 23(2):47-51. Depto Epidemiologia e Saúde Pública, Instituto de Veterinária, UFRRJ, Seropédica, RJ 23890-000, Brazil. Estudou-se o perfil de susceptibilidade a antimicrobianos em cocos Gram-positivos catalase negativos (21 amostras de Lactococcus garvieae e 6 de Enterococcus gallinarum), isoladas do leite de fêmeas com mastite subdínica e pertencentes a uma população composta por seis rebanhos bubalinos localizados no Estado do Rio de Janeiro. O teste utilizado foi o da difusão de discos em agar Müller Hinton, segundo recomendações do National Committee for Clinicai Laboratory Standards - NCCLS, tendo sido testados discos com ampicilina (10mg), cefalotina (30mg), cefotaxima (30mg), cefoxitina (30mg), cloranfenicol (30mg), eritromicina (15mg), gentamicina (10mg), nitrofurantoína (300mg), norfloxacina (10mg), penicilina (10 UI), tetracidina (30mg) e vancomicina (30mg). Os resultados evidenciaram que em se tratando de Lactococcus garvieae, o antimicrobiano mais eficiente foi o nitrofurantoína com 85,71% de sensibilidade, seguido da cefotaxima (61,90%), vancomicina (52,38%), norfloxacina (47,62%) e cefalotina (47,62%). A maior resistência foi desenvolvida frente a penicilina e ampicilina, com 95,24% de resistência para os dois antimicrobianos testados. O perfil de susceptibilidade desenvolvido pelas amostras de Enterococcus gallinarum, mostrou baixa sensibilidade frente aos antimicrobianos testados, onde os maiores índices foram observados frente eritromicina e gentamicina, com 33,34% de sensibilidade para ambos; quanto à resistência desenvolvida, foi possível observar 100% de resistência com relação a vancomicina e tetraciclina, seguindo-se cloranfenicol, penicilina, ampicilina, cefoxitina, cefalotina, cefotaxima, norfloxacina e nitrofurantoína, todas evidenciando uma resistência de 83,33% das amostras testadas.

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.