PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Esposto E.M., Silva W.C.P., Reis C.M.F., Reis E.M.F., Ribeiro R.V., Rodrigues D.P. & Lázaro N.S. 2007. [Bacterial enteropathogens from fishes of a nutrient recycle system and its ecosystem.] Enteropatógenos bacterianos em peixes criados em uma estação de reciclagem de nutrientes e no ecossistema relacionado. Pesquisa Veterinária Brasileira 27(4):144-148. Departamento de Bacteriologia, Instituto Oswaldo Cruz, FIOCruz, Av. Brasil 4365, Pav. Rocha Lima, Manguinhos, Rio de Janeiro, RJ 21040-360, Brazil. E- mail: nslazaro@ioc.fiocruz.br The presence of bacterial enteropathogens from fishes of a nutrient recycle system from a Experimental Station in Petropolis, RJ, was evaluated in 72 samples from april 2000 to july 2001 Simultaneously was collected the mud used as organic manure and poultry beds localized next to the tanks. The isolation procedures included preenrichment in Peptone Water followed by enrichment with alcaline Peptone Water (pH 8.4-8.6), and streaked onto GSP Agar for the isolation of Aeromonas spp. and Plesiomonas shigelloides. For enteropathogenic bacteria of the Enterobacteriaceae family, 1ml samples were transferred for enrichment in Rappaport- Vassiladis broth and Kauffmann Tetrathionate Broth followed by streak onto Hektoen Enteric Agar and Salmonella-Shigella Agar. Simultaneously at each visit samples of water from fish and macrophyte tanks were collected for monitoring faecal coliforms (MPN) using A1 medium. Among the 116 isolates, Aeromonas spp. were the most frequent (67.2%) with 9 species (A. veronii, biogroup sobria, A. hidrophila, A. sobria, A. trota, A. eucrenophila, A. veronii biog. veronii, A. media, A. cavia and A. jandaei), followed by Edwardsiella tarda (16.4%), Plesiomonas shigelloides (12.9%) and Salmonella spp. (3.4%). The NMP of fecal coliforms showed higher values in the fish tanks (>1800/100ml).

• Enteropathogens wastewater fish farming

Abstract in Portuguese:

ABSTRACT.- Esposto E.M., Silva W.C.P., Reis C.M.F., Reis E.M.F., Ribeiro R.V., Rodrigues D.P. & Lázaro N.S. 2007. [Bacterial enteropathogens from fishes of a nutrient recycle system and its ecosystem.] Enteropatógenos bacterianos em peixes criados em uma estação de reciclagem de nutrientes e no ecossistema relacionado. Pesquisa Veterinária Brasileira 27(4):144-148. Departamento de Bacteriologia, Instituto Oswaldo Cruz, FIOCruz, Av. Brasil 4365, Pav. Rocha Lima, Manguinhos, Rio de Janeiro, RJ 21040-360, Brazil. E- mail: nslazaro@ioc.fiocruz.br The presence of bacterial enteropathogens from fishes of a nutrient recycle system from a Experimental Station in Petropolis, RJ, was evaluated in 72 samples from april 2000 to july 2001 Simultaneously was collected the mud used as organic manure and poultry beds localized next to the tanks. The isolation procedures included preenrichment in Peptone Water followed by enrichment with alcaline Peptone Water (pH 8.4-8.6), and streaked onto GSP Agar for the isolation of Aeromonas spp. and Plesiomonas shigelloides. For enteropathogenic bacteria of the Enterobacteriaceae family, 1ml samples were transferred for enrichment in Rappaport- Vassiladis broth and Kauffmann Tetrathionate Broth followed by streak onto Hektoen Enteric Agar and Salmonella-Shigella Agar. Simultaneously at each visit samples of water from fish and macrophyte tanks were collected for monitoring faecal coliforms (MPN) using A1 medium. Among the 116 isolates, Aeromonas spp. were the most frequent (67.2%) with 9 species (A. veronii, biogroup sobria, A. hidrophila, A. sobria, A. trota, A. eucrenophila, A. veronii biog. veronii, A. media, A. cavia and A. jandaei), followed by Edwardsiella tarda (16.4%), Plesiomonas shigelloides (12.9%) and Salmonella spp. (3.4%). The NMP of fecal coliforms showed higher values in the fish tanks (>1800/100ml).

Abstract in English:

ABSTRACT.- Vieira A.C.S., Afonso J.A.B. & Mendonça C.L. 2007. [Ruminal fluid characteristics of Santa Inês sheep under pasture conditions in the State of Pernambuco.] Características do fluído ruminal de ovinos Santa Inês criados extensivamente em Pernambuco. Pesquisa Veterinária Brasileira 27(3):110-114. Clínica de Bovinos, Universidade Federal Rural de Pernambuco (UFRPE), Cx. Postal 152, Garanhuns, PE 55292-901, Brazil. E-mail: acynnara@gmail.com The objective of this study was to determine normal standards to ruminal fluid characteristics of Santa Inês sheep under pasture conditions in the State of Pernambuco. Fifty samples were collected, using an esophageal tube, during winter (rainy season) and summer (dry season). The predominant colors were an olive green in the rainy season and a nut-brown in the dry season. The smell was aromatic, but was stronger in winter. A slight viscous consistence was found in most of the samples, with a greater proportion in winter. The sedimentation and flotation time was 6.73min (±1.63) in the rainy period and 3.15min (±0.72) in the dry period. In the biochemical tests, the average values found in winter and summer were, respectively: pH 6.76±0.21 and 6,59±0.14; methylene blue reduction, 3.20min (±0.76) and 7.76min (±3.00); chloride, 28.14±4,16mEq/L and 24.97±5.65mEq/L; acidity, 21.90±4.38UC and 13.68±2.97UC. Ruminal microbiotic analyses revealed abundant and moderate density of protozoa in winter and summer, respectively. The motility was very active and there were almost 90% of live protozoa in both seasons. Protozoa numbers were 425.373±217.258/mL in winter and 155.375±83.113/mL in summer. There was a mixed population of bacteria with prevalence of Gram-negative forms in both seasons.

• Ruminal fluid sheep bacteria protozoa

Abstract in Portuguese:

ABSTRACT.- Vieira A.C.S., Afonso J.A.B. & Mendonça C.L. 2007. [Ruminal fluid characteristics of Santa Inês sheep under pasture conditions in the State of Pernambuco.] Características do fluído ruminal de ovinos Santa Inês criados extensivamente em Pernambuco. Pesquisa Veterinária Brasileira 27(3):110-114. Clínica de Bovinos, Universidade Federal Rural de Pernambuco (UFRPE), Cx. Postal 152, Garanhuns, PE 55292-901, Brazil. E-mail: acynnara@gmail.com The objective of this study was to determine normal standards to ruminal fluid characteristics of Santa Inês sheep under pasture conditions in the State of Pernambuco. Fifty samples were collected, using an esophageal tube, during winter (rainy season) and summer (dry season). The predominant colors were an olive green in the rainy season and a nut-brown in the dry season. The smell was aromatic, but was stronger in winter. A slight viscous consistence was found in most of the samples, with a greater proportion in winter. The sedimentation and flotation time was 6.73min (±1.63) in the rainy period and 3.15min (±0.72) in the dry period. In the biochemical tests, the average values found in winter and summer were, respectively: pH 6.76±0.21 and 6,59±0.14; methylene blue reduction, 3.20min (±0.76) and 7.76min (±3.00); chloride, 28.14±4,16mEq/L and 24.97±5.65mEq/L; acidity, 21.90±4.38UC and 13.68±2.97UC. Ruminal microbiotic analyses revealed abundant and moderate density of protozoa in winter and summer, respectively. The motility was very active and there were almost 90% of live protozoa in both seasons. Protozoa numbers were 425.373±217.258/mL in winter and 155.375±83.113/mL in summer. There was a mixed population of bacteria with prevalence of Gram-negative forms in both seasons.

Abstract in English:

Zafalon L.F., Nader Filho A., Oliveira J.V. & Resende F.D. 2005. [Electrical conductivity and chloride concentration of milk as auxiliary diagnostic methods in bovine subclinical mastitis.] Comportamento da condutividade elétrica e do conteúdo de cloretos do leite como métodos auxiliares de diagnóstico na mastite subclínica bovina. Pesquisa Veterinária Brasileira 25(3):159-163. Depto Medicina Veterinária Preventiva e Reprodução Animal, FACVJ, Unesp - Campus de Jaboticabal, Via de Acesso Prof. Paulo Donato Castellane s/no, Jaboticabal, SP 14884-900, Brazil. E-mail: zafalon@iz.sp.gov.br Electrical conductivity measured by a hand-held meter and chloride concentration of milk were studied as auxiliary methods for diagnosis of bovine subclinical mastitis in the identification of affected mammary quarters where Staphylococcus aureus and Corynebacterium sp were later isolated. Tests were made during 2 years in Holstein cows of a dairy farm producing type C milk, where milking was performed once a day. Sensitivities of electrical conductivity and chloride concentration tests from mammary quarters, where Corynebacterium sp was isolated (65.3% and 78.3%, respectively), were superior to the found in mammary quarters where S. aureus was identified (55.4% and 68.2%, respectively). The efficacies of the two diagnostic tests were similar. Statistical significance was demonstrated with regression analysis of both tests of healthy mammary quarters and subclinical mastitis quarters infected with Staphylococcus aureus.

• Bovine subclinical mastitis electric conductivity chloride level Staphylococcus aureus Corynebacterium sp.

Abstract in Portuguese:

Zafalon L.F., Nader Filho A., Oliveira J.V. & Resende F.D. 2005. [Electrical conductivity and chloride concentration of milk as auxiliary diagnostic methods in bovine subclinical mastitis.] Comportamento da condutividade elétrica e do conteúdo de cloretos do leite como métodos auxiliares de diagnóstico na mastite subclínica bovina. Pesquisa Veterinária Brasileira 25(3):159-163. Depto Medicina Veterinária Preventiva e Reprodução Animal, FACVJ, Unesp - Campus de Jaboticabal, Via de Acesso Prof. Paulo Donato Castellane s/no, Jaboticabal, SP 14884-900, Brazil. E-mail: zafalon@iz.sp.gov.br Electrical conductivity measured by a hand-held meter and chloride concentration of milk were studied as auxiliary methods for diagnosis of bovine subclinical mastitis in the identification of affected mammary quarters where Staphylococcus aureus and Corynebacterium sp were later isolated. Tests were made during 2 years in Holstein cows of a dairy farm producing type C milk, where milking was performed once a day. Sensitivities of electrical conductivity and chloride concentration tests from mammary quarters, where Corynebacterium sp was isolated (65.3% and 78.3%, respectively), were superior to the found in mammary quarters where S. aureus was identified (55.4% and 68.2%, respectively). The efficacies of the two diagnostic tests were similar. Statistical significance was demonstrated with regression analysis of both tests of healthy mammary quarters and subclinical mastitis quarters infected with Staphylococcus aureus.

Abstract in English:

Simionatto S., Vaz E.K., Michelon A., Seixas F.K., Dellagostin O.A. 2005. [Development and evaluation of new strategies for immunization against swine colibacillosis.] Desenvolvimento e avaliação de novas estratégias de imunização contra colibacilose suína. Pes-quisa Veterinária Brasileira 25(2):84-90. Laboratório de Biologia Molecular, Centro de Bio-tecnologia, UFPel, Campus Capão do Leão, Cx. Postal 354, Pelotas, RS 96010-900, Brazil. E-mail: ssimionatto@bol.com.br Swine colibacillosis caused by enterotoxigenic Escherichia coli remains one of the main sanitary problems in pig farms. The recombinant DNA technology offers the possibility of developing new immunization strategies. This paper describes the development of a subunit vaccine through the expression and purification of the E. coli K88 FaeC fimbrial protein. The gene that codes for this antigen was amplified by PCR and cloned into an E. coli expression vector fused to a 6X histidine tag. The recombinant protein was purified by affinity chromatography and used for mice immunization. In parallel, the same gene was cloned into an eucariotic expression vector with the addition of the Kozak sequence for improving translation of this gene in muscle cells. The resulting plasmid named pUP310 was purified in large scale and used to immunize mice. The immune response afforded by both forms of immunization was monitored by ELISA. There was an immune response in mice inoculated with pUP310 and purified FaeC. It was possible to detect anti-FaeC antibodies 42 days after the first inoculation. The antibody titer increased with time, being still detectable 7 months after the first inoculation. It is concluded that recombinant FaeC and pUP310 are potential tools for immunization of swine against E. coli K88.

• Swine colibacillosis FaeC Escherichia coli DNA vaccine recombinant vaccine.

Abstract in Portuguese:

Simionatto S., Vaz E.K., Michelon A., Seixas F.K., Dellagostin O.A. 2005. [Development and evaluation of new strategies for immunization against swine colibacillosis.] Desenvolvimento e avaliação de novas estratégias de imunização contra colibacilose suína. Pes-quisa Veterinária Brasileira 25(2):84-90. Laboratório de Biologia Molecular, Centro de Bio-tecnologia, UFPel, Campus Capão do Leão, Cx. Postal 354, Pelotas, RS 96010-900, Brazil. E-mail: ssimionatto@bol.com.br Swine colibacillosis caused by enterotoxigenic Escherichia coli remains one of the main sanitary problems in pig farms. The recombinant DNA technology offers the possibility of developing new immunization strategies. This paper describes the development of a subunit vaccine through the expression and purification of the E. coli K88 FaeC fimbrial protein. The gene that codes for this antigen was amplified by PCR and cloned into an E. coli expression vector fused to a 6X histidine tag. The recombinant protein was purified by affinity chromatography and used for mice immunization. In parallel, the same gene was cloned into an eucariotic expression vector with the addition of the Kozak sequence for improving translation of this gene in muscle cells. The resulting plasmid named pUP310 was purified in large scale and used to immunize mice. The immune response afforded by both forms of immunization was monitored by ELISA. There was an immune response in mice inoculated with pUP310 and purified FaeC. It was possible to detect anti-FaeC antibodies 42 days after the first inoculation. The antibody titer increased with time, being still detectable 7 months after the first inoculation. It is concluded that recombinant FaeC and pUP310 are potential tools for immunization of swine against E. coli K88.

Abstract in English:

Hofer E. & Reis C.M.F. 2005. [Species and serovars of Listeria isolated from sick and clinically healthy animals in Brazil.] Espécies e sorovares de Listeria isolados de animais doentes e portadores no Brasil. Pesquisa Veterinária Brasileira 25(2):79-83. Laboratório de Zoonoses Bacterianas, Depto Bacteriologia, Instituto Oswaldo Cruz/FIOCRUZ, Rio de Janeiro, RJ 21045-900, Brazil. E-mail: ehofer@ioc.fiocruz.br Two hundred fourty-six strains of the genus Listeria were isolated from sick and clinically healthy animals, collected in three different regions of Brazil during 1971-2000. About 88.2% (217 cultures) yielded Listeria species from faecal specimens of healthy cattle and 29 strains (11.7%) were isolated from sick animals: 15 (6.0%) from central nervous system (CNS) and 14(5.6%) were from otherwise sterile sites. Phenotyping techniques were used to characterize the Listeria isolates. The commonest were L. innocua 6a and non-typable (140/56.9%), L. monocytogenes 4a (37/15.0%) and 4b (22/8.9%), originated mainly from stools of healthy cattle. From sick animals the predominant species and serovars were L. monocytogenes 4b (14/5.6%), and the higher incidence was observed in ruminants (12/4.8%) and 8/3.2% of the serovar 1a from other animal species (rodents and canines) mainly isolated from CNS samples.

• Listeria species serovars sick and healthy animals.

Abstract in Portuguese:

Hofer E. & Reis C.M.F. 2005. [Species and serovars of Listeria isolated from sick and clinically healthy animals in Brazil.] Espécies e sorovares de Listeria isolados de animais doentes e portadores no Brasil. Pesquisa Veterinária Brasileira 25(2):79-83. Laboratório de Zoonoses Bacterianas, Depto Bacteriologia, Instituto Oswaldo Cruz/FIOCRUZ, Rio de Janeiro, RJ 21045-900, Brazil. E-mail: ehofer@ioc.fiocruz.br Two hundred fourty-six strains of the genus Listeria were isolated from sick and clinically healthy animals, collected in three different regions of Brazil during 1971-2000. About 88.2% (217 cultures) yielded Listeria species from faecal specimens of healthy cattle and 29 strains (11.7%) were isolated from sick animals: 15 (6.0%) from central nervous system (CNS) and 14(5.6%) were from otherwise sterile sites. Phenotyping techniques were used to characterize the Listeria isolates. The commonest were L. innocua 6a and non-typable (140/56.9%), L. monocytogenes 4a (37/15.0%) and 4b (22/8.9%), originated mainly from stools of healthy cattle. From sick animals the predominant species and serovars were L. monocytogenes 4b (14/5.6%), and the higher incidence was observed in ruminants (12/4.8%) and 8/3.2% of the serovar 1a from other animal species (rodents and canines) mainly isolated from CNS samples.

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:

Assis R.A., Lobato F.C.F., Serakides R., Santos R.L., Dias G.R.C., Nascimento R.A.P., Abreu V.L.V, Parreiras P.M. & Uzal F.A. 2005. Immunohistochemical detection of Clostridia species in paraffin-embedded tissues of experimentally inoculated guinea pigs. Pesquisa Veterinária Brasileira 25(1):4-8. Departamento de Medicina Veterinária Preventiva, Escola de Veterinária, Universidade Federal de Minas Gerais, Av. Presidente Antônio Carlos 6627, Cx. Postal 567, Belo Horizonte, MG 30123-970, Brazil. E-mail: assisra@rwnet.com.br Blackleg is caused by Clostridium chauvoei, whereas malignant oedema is caused by C. chauvoei, C. septicum, C. sordellii, C. perfringens type A, and/or C. novyi type A. Anti-C. chauvoei, anti-C. septicum, anti-C. sordellii and anti-C. novyi type A polyclonal antibodies were produced in rabbits and purified in a column of DEAE-cellulose. Aliquots of the antisera were conjugated with fluorescein isothiocyanate and the remaining was used for the streptavidin biotin peroxidase technique (SBPT). SBPT was standardized to detect C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs. SBPT was compared to a fluorescent antibody technique (FAT). Sections and smears of muscle from inoculation area (MIA), heart, liver, spleen and kidney, were obtained for both SBPT and FAT. Cross-reactions between the different Clostridial species were not observed. C. chauvoei and C. septicum were detected in all specimens from the animals inoculated with these microorganisms, while only sections of muscle obtained from all the animals inoculated with C. sordellii and C. novyi type A were positive. The same results observed by the SBPT, were obtained on tissue smears of these microorganisms stained by the FAT. The results indicate that SBPT is suitable for detection of C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs.

• Blackleg malignant oedema Clostridia guinea pigs fluorescent antibody technique streptavidin biotin peroxidase technique.

Abstract in Portuguese:

Assis R.A., Lobato F.C.F., Serakides R., Santos R.L., Dias G.R.C., Nascimento R.A.P., Abreu V.L.V, Parreiras P.M. & Uzal F.A. 2005. Immunohistochemical detection of Clostridia species in paraffin-embedded tissues of experimentally inoculated guinea pigs. Pesquisa Veterinária Brasileira 25(1):4-8. Departamento de Medicina Veterinária Preventiva, Escola de Veterinária, Universidade Federal de Minas Gerais, Av. Presidente Antônio Carlos 6627, Cx. Postal 567, Belo Horizonte, MG 30123-970, Brazil. E-mail: assisra@rwnet.com.br Blackleg is caused by Clostridium chauvoei, whereas malignant oedema is caused by C. chauvoei, C. septicum, C. sordellii, C. perfringens type A, and/or C. novyi type A. Anti-C. chauvoei, anti-C. septicum, anti-C. sordellii and anti-C. novyi type A polyclonal antibodies were produced in rabbits and purified in a column of DEAE-cellulose. Aliquots of the antisera were conjugated with fluorescein isothiocyanate and the remaining was used for the streptavidin biotin peroxidase technique (SBPT). SBPT was standardized to detect C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs. SBPT was compared to a fluorescent antibody technique (FAT). Sections and smears of muscle from inoculation area (MIA), heart, liver, spleen and kidney, were obtained for both SBPT and FAT. Cross-reactions between the different Clostridial species were not observed. C. chauvoei and C. septicum were detected in all specimens from the animals inoculated with these microorganisms, while only sections of muscle obtained from all the animals inoculated with C. sordellii and C. novyi type A were positive. The same results observed by the SBPT, were obtained on tissue smears of these microorganisms stained by the FAT. The results indicate that SBPT is suitable for detection of C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs.

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:

Portiansky E.L., Quiroga M.A., Machuca M.A. & Perfumo C.J. 2004. Mycoplasma suis in naturally infected pigs: an ultrastructural and morphometric study. Pesquisa Veterinária Brasileira 24(1):1-5. Instituto de Patología, Faculdad de Ciencias Veterinarias, Universidad Nacional de La Plata, Calle 60 y 118, C.C. 296, (1900) La Plata, Buenos Aires, Argentina. E-mail: elporti@fcv.unlp.edu.ar Swine eperythrozoonosis is a haemotrophic disease caused by Eperythrozoon suis, actually called Mycoplasma suis, an extracellular bacterial organism that apparently adheres to pig erythrocyte membrane, inducing its deformation and damage. Since little is known about the ultrastructural and morphometrical aspects of this microorganism, the present work aimed to deal with these issues. The ultrastructural study revealed the presence of structures corresponding to tubules disseminated throughout the soma of M. suis. A variable separation between the microorganism membrane and that of the erythrocyte was also observed. The structural and positional attitude of M. suis could allow speculation about its mechanism of action.

• Eperythrozoonosis Mycoplasma suis ultrastructure morphometry pathogenesis.

Abstract in Portuguese:

Portiansky E.L., Quiroga M.A., Machuca M.A. & Perfumo C.J. 2004. Mycoplasma suis in naturally infected pigs: an ultrastructural and morphometric study. Pesquisa Veterinária Brasileira 24(1):1-5. Instituto de Patología, Faculdad de Ciencias Veterinarias, Universidad Nacional de La Plata, Calle 60 y 118, C.C. 296, (1900) La Plata, Buenos Aires, Argentina. E-mail: elporti@fcv.unlp.edu.ar Swine eperythrozoonosis is a haemotrophic disease caused by Eperythrozoon suis, actually called Mycoplasma suis, an extracellular bacterial organism that apparently adheres to pig erythrocyte membrane, inducing its deformation and damage. Since little is known about the ultrastructural and morphometrical aspects of this microorganism, the present work aimed to deal with these issues. The ultrastructural study revealed the presence of structures corresponding to tubules disseminated throughout the soma of M. suis. A variable separation between the microorganism membrane and that of the erythrocyte was also observed. The structural and positional attitude of M. suis could allow speculation about its mechanism of action.

Abstract in English:

ABSTRACT.- Barbosa]. D., Oliveira C. M. C., Tokarnia C. H. & Riet-Correa E 2003. [Comparison of the sensibility of cattle and buffaloes to poisoning by Palicourea marcgravii (Rubiaceae).] Comparação da sensibilidade de bovinos e búfalos à intoxicação por Palicourea marcgravii (Rubiaceae). Pesquisa Veterinária Brasileira 23(4): 167-172. Escola de Medicina Veterinária, Campus Castanhal, Universidade Federal do Pará, Rua Maximino Porpino 1000, Castanhal, PA 68743- 080, Brazil. E-mail: diomedes@ufpa.br In order to verify if buffaloes are more resistant than cattle to poisoning by Palicourea marcgravii, the plant was given per os simultaneously to buffaloes and cattle. Seven buffaloes and three head of cattle were used. Doses of 0.5, 1.0 and 2.0 g/kg did not cause signs of poisoning in buffaloes; but doses of 3.0, 4.0 and 6.0 g/kg caused death. In cattle 0.25 g/kg did not cause symptoms; but 0.5 and 2.0 g/kg were lethal. The influence of exercise on the onset of symptoms, the time which elapsed from the beginning of the administration of the plant to the appearance of symptoms, and the symptoms themselves, were similar in the two animal species. The clinical course was longer in buffaloes; whilst in cattle the period between the onset of severe symptoms and death lasted 9 to 17 minutes; in buffaloes it varied from 10 minutes to 1 hour 28 minutes. It can be concluded, that buffaloes are about six times more resistant than cattle to the toxic action of P. maracgravii. The lower incidence of deaths in buffaloes by poisonous plants in the Amazon region is, at least in part, due the lower sensibility of buffaloes to poisoning by this plant. Another factor responsible for the smaller number of buffaloes which die due to plant poisoning in the Amazon region, is possibly that buffaloes prefer the Jowlands, e.g. temporarily flooded areas, which are the habitat of Arrabidaea bilabiata, the second most important poisonous plant of the Amazon region. This plant is less poisonous than P. marcgravii, and its habitat are the higher not flooded areas. So it would be safer, in order to diminish losses, to breed buffaloes instead of cattle in areas where P. marcgravii occurs. The reason for the greater resistance of buffaloes should be further investigated as a possible prophylactic measure to avoid poisoning by P. marcgravii in cattle. On the other hand, Australian investigators modified genetically the ruminai bacteria Butyrivibrio fibrisolvens by introducing a gene, isolated from a Moraxella sp, which codifies a dehalogenase, capable of hydrolyzing fluoroacetate, the poisonous principle of P. marcgravii. Transferring the genetically modified B. fibrisolvens into the rumen of animal, which consume plants that contain fluoroacetate, would be a viable method to control that poisoning through a ruminal detoxifying process for the toxin. Through preliminary contacts, the director of the consorcium in Australia, responsible for the modification of the bacteria, declared that he is interested to sell the technology to our country, but it would be necessary to know if it is permitted to introduce into Brazil the genetically modified bacteria. If allowed, it would be necessary to perform investigations on the viability and the methodology for using this bacteria to control P. marcgravii poisoning in our country.

• Poisonous plants plant poisoning Palicourea marcgravii Rubiaceae cattle buffaloes prophylaxis Butyrivibrio fibrisolvens genetically modified bactéria Intoxicação por planta Rubiaceae bovinos búfalos profilaxia bactérias geneticamente modificadas.

Abstract in Portuguese:

RESUMO.- Barbosa]. D., Oliveira C. M. C., Tokarnia C. H. & Riet-Correa E 2003. [Comparison of the sensibility of cattle and buffaloes to poisoning by Palicourea marcgravii (Rubiaceae).] Comparação da sensibilidade de bovinos e búfalos à intoxicação por Palicourea marcgravii (Rubiaceae). Pesquisa Veterinária Brasileira 23(4): 167-172. Escola de Medicina Veterinária, Campus Castanhal, Universidade Federal do Pará, Rua Maximino Porpino 1000, Castanhal, PA 68743- 080, Brazil. E-mail: diomedes@ufpa.br O presente estudo foi realizado para verificar se búfalos são mais resistentes do que bovinos à ação tóxica de Palicourea marcgravii, mediante a administração da planta por via oral, simultaneamente, a bovinos e a búfalos. Foram usados sete búfalos e três bovinos. Verificou-se que em búfalos doses de 0,5 g/kg, 1,0 g/kg e 2,0 g/kg não causaram sintomas de intoxicação. As doses de 3,0 g/kg, 4,0 g/kg e 6,0 g/kg causaram a morte dos búfalos. Em bovinos, a dose de 0,25g/kg não causou sinais clínicos de intoxicação, enquanto que doses de 0,5 g/kg e 2,0 g/kg causaram a morte. A influência do exercício sobre o aparecimento dos sintomas, o prazo decorrido desde o começo da administração da planta até o início de sintomas, e os próprios sintomas, foram semelhantes nas duas espécies animais. O curso clínico foi mais longo nos búfalos. Enquanto nos bovinos o período entre o aparecimento dos sintomas graves e a morte foi de 9 a 17 minutos, nos búfalos variou de 10 minutos a 1 hora e 28 minutos. Pode se concluir que os bubalinos são aproximadamente seis vezes mais resistentes do que os bovinos à ação tóxica de P. marcgravii. O menor índice de mortes pela ação de plantas tóxicas na Amazônia em búfalos é, pelo menos em parte, devido à maior resistência do búfalo à intoxicação por essa planta. Outro fator responsável pelo menor número de mortes em búfalos pela intoxicação por plantas, na Amazônia, poderia ser que os búfalos preferem a várzea, que é o habitat de Arrabidaea bilabiata, a segunda planta tóxica mais importante da Amazônia, menos tóxica do que P. marcgravii, e com habitat na terra firme. Em áreas onde ocorre P. marcgravii seria mais prudente, para diminuir os prejuízos, criar búfalos em lugar de bovinos. A causa dessa maior resistência do búfalo merece ser investigada para a eventual elaboração de métodos profiláticos da intoxicação por P. marcgravii em bovinos. Por outro lado, pesquisadores australianos modificaram geneticamente a bactéria ruminal Butyrivibrio fibrisolvens, mediante a introdução de um gene, isolado de Moraxella sp, que codifica uma dehalogenase, capaz de hidro-lizar fluoroacetato. A transferência de B.fibrisolvens geneticamente modificado para o rúmen de animais ingerindo plantas que contêm fluoroacetato, como é o caso de P. marcgravii, seria um método viável para o controle da intoxicação mediante a detoxificação ruminal do princípio ativo. Em contatos preliminares o diretor responsável do consórcio na Austrália responsável pela modificação da bactéria, declarou o interesse em vender a tecnologia ao nosso país, porém seria necessário saber se é possível importar essa bactéria geneticamente modificada no Brasil. Caso positivo, seria indispensável realizar pesquisas sobre a viabilidade e a metodologia para o uso dessa bactéria em nosso meio.