PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Melo E.S.P., Araújo F.R., Ramos C.A.N., Soares C.O., Rosinha G.M.S., Elisei C. & Madruga C.R. 2007. [ELISA based on recombinant truncated MSP5 for detection of antibodies against Anaplasma marginale in cattle.] ELISA com MSP5 recombinante truncada para detecção de anticorpos contra Anaplasma marginale em bovinos. Pesquisa Veterinária Brasileira 27(7):301-306. Embrapa Gado de Corte, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. E-mail: flabio@cnpgc.embrapa.br The objective of this study was the production and solubilization of recombinant truncated MSP5 of Anaplasma marginale and the evaluation of its performance in an enzyme-linked immunosorbent assay (ELISA), to detect antibodies against the rickettsia in cattle. The fragment of msp5 gene, except the hydrophobic N-terminal region, was amplified by PCR, cloned in pTrcHis-TOPO plasmid and expressed in Escherichia coli. Solubilization of the recombinant protein was evaluated in different pHs and concentrations of urea. The sensibility and specificity of the assay were evaluated with 66 sera from cattle experimentally-infected and 96 sera from cattle free of A. marginale defined by polymerase chain reaction for msp5 gene. Serum samples from 1,666 cattle from Brazil - states of Rio Grande do Sul (73), Mato Grosso do Sul (91), Pernambuco (86), Bahia (314) and Minas Gerais (267), Uruguay (32) and Costa Rica (803), were tested by ELISAs with recombinant truncated MSP5 and with recombinant MSP1a, and the agreement between both ELISAs was calculated. ELISA with recombinant truncated MSP5 protein detected infected animals with sensibility of 96.97% and specificity of 100%. In cattle experimentally-infected, the ELISA detected antibodies from the 12th day post-infection (DPI) to the end of the experiment, at the 37th DPI. The agreement between the ELISAs with truncated MSP5 and MSP1a antigens was 95.67%, with a kappa index of 0.81. Disagreement results showed significative difference (p <0.001). Antibodies for A. marginale were detected in animals of the all the region analyzed. The ELISA with recombinant truncated MSP5 showed a good performance in ELISA for detention of antibodies against A. marginale, with high sensitivity and specificity, representing an important tool for the diagnosis of anaplasmose bovine in epidemiological studies.

• Anaplasma marginale ELISA MSP5 solubilization recombinant protein

Abstract in Portuguese:

ABSTRACT.- Melo E.S.P., Araújo F.R., Ramos C.A.N., Soares C.O., Rosinha G.M.S., Elisei C. & Madruga C.R. 2007. [ELISA based on recombinant truncated MSP5 for detection of antibodies against Anaplasma marginale in cattle.] ELISA com MSP5 recombinante truncada para detecção de anticorpos contra Anaplasma marginale em bovinos. Pesquisa Veterinária Brasileira 27(7):301-306. Embrapa Gado de Corte, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. E-mail: flabio@cnpgc.embrapa.br The objective of this study was the production and solubilization of recombinant truncated MSP5 of Anaplasma marginale and the evaluation of its performance in an enzyme-linked immunosorbent assay (ELISA), to detect antibodies against the rickettsia in cattle. The fragment of msp5 gene, except the hydrophobic N-terminal region, was amplified by PCR, cloned in pTrcHis-TOPO plasmid and expressed in Escherichia coli. Solubilization of the recombinant protein was evaluated in different pHs and concentrations of urea. The sensibility and specificity of the assay were evaluated with 66 sera from cattle experimentally-infected and 96 sera from cattle free of A. marginale defined by polymerase chain reaction for msp5 gene. Serum samples from 1,666 cattle from Brazil - states of Rio Grande do Sul (73), Mato Grosso do Sul (91), Pernambuco (86), Bahia (314) and Minas Gerais (267), Uruguay (32) and Costa Rica (803), were tested by ELISAs with recombinant truncated MSP5 and with recombinant MSP1a, and the agreement between both ELISAs was calculated. ELISA with recombinant truncated MSP5 protein detected infected animals with sensibility of 96.97% and specificity of 100%. In cattle experimentally-infected, the ELISA detected antibodies from the 12th day post-infection (DPI) to the end of the experiment, at the 37th DPI. The agreement between the ELISAs with truncated MSP5 and MSP1a antigens was 95.67%, with a kappa index of 0.81. Disagreement results showed significative difference (p <0.001). Antibodies for A. marginale were detected in animals of the all the region analyzed. The ELISA with recombinant truncated MSP5 showed a good performance in ELISA for detention of antibodies against A. marginale, with high sensitivity and specificity, representing an important tool for the diagnosis of anaplasmose bovine in epidemiological studies.

Abstract in English:

ABSTRACT.- Ferrigno C.R.A.,Della Nina M.I. & Fantoni D.T. 2007. [A comparative study of osteosynthesis with plates and plates associated with grafts of bone morphogenic proteins (Gen-Techâ) in distal radio-ulna fractures in dogs with less than 6 quilograms.] Estudo comparativo entre as osteossínteses com placas e osteossínteses com placas associadas a enxertos de proteína morfogenética óssea (Gen-Techâ) em fraturas distais de rádio-ulna em cães com menos de 6 quilos. Pesquisa Veterinária Brasileira 27(2):65-69. Departamento de Cirurgia do Hospital Veterinário, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Orlando Marques de Paiva 87, São Paulo, SP 05508-900. E-mail: cassioaf@usp.br It is well known that bone morphogenic proteins (BMP) cause osteogenesis, yet clinical research must be performed in order to really show their benefits. Animals weighing less than 6 kg show well known difficulties regarding radius and ulna fracture repair mainly with bone non-union, due to poor vascularization of the distal portion of the radius. Therefore this study aimed to compare the velocity of bone callus formation in the treatment of fracture repair with plates and screws alone or with plates and screws plus BMP. Thirty three dogs with radius and ulna fractures were distributed into two groups, where animals of the control group received the conservative treatment performed with screws and plates alone, whilst the other group received the conservative treatment and BMP. The time of bone callus formation was evaluated comparatively through radiographic exams 30, 60, 90, 120, 180 and 210 days after the surgical procedure. Animals treated with BMP showed a healing time of 32±15 days, which was significantly different (p <0.001) from the control group which required 127±34 days. With the results obtained, it can be concluded that the distal radio-ulna fractures of dogs weighing less than 6 kg suffered a significant reduction of the bone callus formation time, which was around 90 days.

• Bone morphogenic proteins BMP bone fractures bone plate dogs

Abstract in Portuguese:

ABSTRACT.- Ferrigno C.R.A.,Della Nina M.I. & Fantoni D.T. 2007. [A comparative study of osteosynthesis with plates and plates associated with grafts of bone morphogenic proteins (Gen-Techâ) in distal radio-ulna fractures in dogs with less than 6 quilograms.] Estudo comparativo entre as osteossínteses com placas e osteossínteses com placas associadas a enxertos de proteína morfogenética óssea (Gen-Techâ) em fraturas distais de rádio-ulna em cães com menos de 6 quilos. Pesquisa Veterinária Brasileira 27(2):65-69. Departamento de Cirurgia do Hospital Veterinário, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Orlando Marques de Paiva 87, São Paulo, SP 05508-900. E-mail: cassioaf@usp.br It is well known that bone morphogenic proteins (BMP) cause osteogenesis, yet clinical research must be performed in order to really show their benefits. Animals weighing less than 6 kg show well known difficulties regarding radius and ulna fracture repair mainly with bone non-union, due to poor vascularization of the distal portion of the radius. Therefore this study aimed to compare the velocity of bone callus formation in the treatment of fracture repair with plates and screws alone or with plates and screws plus BMP. Thirty three dogs with radius and ulna fractures were distributed into two groups, where animals of the control group received the conservative treatment performed with screws and plates alone, whilst the other group received the conservative treatment and BMP. The time of bone callus formation was evaluated comparatively through radiographic exams 30, 60, 90, 120, 180 and 210 days after the surgical procedure. Animals treated with BMP showed a healing time of 32±15 days, which was significantly different (p <0.001) from the control group which required 127±34 days. With the results obtained, it can be concluded that the distal radio-ulna fractures of dogs weighing less than 6 kg suffered a significant reduction of the bone callus formation time, which was around 90 days.

Abstract in English:

Lima V.M.F., Biazzono L., Silva A.C., Correa A.P.F.L. & Luvizotto M.C.R. 2005. Serological diagnosis of visceral leishmaniasis by an enzyme immunoassay using protein A in naturally infected dogs. Pesquisa Veterinária Brasileira 25(4):215-218. Departamento de Clínica, Cirurgia e Reprodução Animal, Faculdade de Medicina Veterinária, Universidade Estadual Paulista, Rua Clóvis Pestana 793, Araçatuba, SP 16050-680, Brazil. E-mail: vmflima@fmva.unesp.br A rapid indirect enzyme-linked immunosorbent assay (ELISA) was developed for measuring antibodies against Leishmania chagasi using total antigen from lysed promastigotes. Fifty symptomatic mixed breed dogs from a region of high incidence of visceral leishmaniasis in Brazil were examined. The results showed that in the positive animals, diagnosed by cytological examination, the ELISA using protein A assay system (mean optical density ± SD / 2.078 ± 0.631) detected more antibodies than the anti-IgG assay (mean optical density ± SD / 1.008 ± 0.437), while in the negative animals, the results by both systems were similar. These results suggest that the ELISA assay using protein A peroxidase conjugated could be useful to detect early infected animals in endemic areas, and thus help to control the spread of the infection.

• Leishmania chagasi visceral leishmaniasis protein A dogs.

Abstract in Portuguese:

Lima V.M.F., Biazzono L., Silva A.C., Correa A.P.F.L. & Luvizotto M.C.R. 2005. Serological diagnosis of visceral leishmaniasis by an enzyme immunoassay using protein A in naturally infected dogs. Pesquisa Veterinária Brasileira 25(4):215-218. Departamento de Clínica, Cirurgia e Reprodução Animal, Faculdade de Medicina Veterinária, Universidade Estadual Paulista, Rua Clóvis Pestana 793, Araçatuba, SP 16050-680, Brazil. E-mail: vmflima@fmva.unesp.br A rapid indirect enzyme-linked immunosorbent assay (ELISA) was developed for measuring antibodies against Leishmania chagasi using total antigen from lysed promastigotes. Fifty symptomatic mixed breed dogs from a region of high incidence of visceral leishmaniasis in Brazil were examined. The results showed that in the positive animals, diagnosed by cytological examination, the ELISA using protein A assay system (mean optical density ± SD / 2.078 ± 0.631) detected more antibodies than the anti-IgG assay (mean optical density ± SD / 1.008 ± 0.437), while in the negative animals, the results by both systems were similar. These results suggest that the ELISA assay using protein A peroxidase conjugated could be useful to detect early infected animals in endemic areas, and thus help to control the spread of the infection.

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

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

Abstract in Portuguese:

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

Abstract in English:

ABSTRACT.- Schenk M.A.M., Mendonça C.L., Madruga C.R., Kohayagawa A. & Araújo F.R. 2001. [Clinical and laboratorial evaluation of Nellore cattle experimentally infected with Trypanosoma vivax] Avaliação clínico and laboratorial de bovinos nelore infectados experimentalmente com Trypanosoma vivax. Pesquisa Veterinária Brasileira 21(4):157-161. Embrapa Gado de Corte, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. In arder to evaluate the clinical-laboratorial alterations, six Neil ore calves were inoculated with 107 Trypanosoma vivax isolated from Poconé region, Mato Grosso, Brazil. The animals were evaluated daily for rectal temperature, packed cell volume (PCV), parasitemia, antibody production, calor of mucous membranes, behavior and appetite. Blood and serum samples for biochemical evaluation for aspartate aminotransferase (AST), alkaline phbsphatase (AF), gamma glutamyltransferase (GGT), cholesterol, urea, creatinine, creatine kinase (CK), calcium, phosphorus and proteinogram were collected on days 4, 8, 12, 16, 23 and 30 post inoculation (DPI). During the following 6 months rectal temperature, PCV and parasitemia were evaluated weekly. T. vivax was evidenced from 1 DPI in all calves and persisted until day 30 in five of six animals. A remarkable decrease (p<0.05) of PCV mean value (25%) was observed on 10 DPI. The animals presented no alterations in their clinical or serum biochemical state during the trial. Seroconversion took place 6 and 8 DPI, and all the animals remained seropositive during the 30 days of experiment. In all the experimental animals the occurrence of T. vivax infection was verified, characterized by the increase of corporal temperature, presente of the blood protozoa and reduction of the globular volume, without altéations in the other variables analyzed. Nellore calves, when experimentally inoculated with T. vivax, are able to establish a balance between host-parasite relationship.

• Trypanosoma vivax calves hematology serum biochemistry proteinogram bezerros hematologia bioquímica sérica proteinograma.

Abstract in Portuguese:

RESUMO.- Schenk M.A.M., Mendonça C.L., Madruga C.R., Kohayagawa A. & Araújo F.R. 2001. [Clinical and laboratorial evaluation of Nellore cattle experimentally infected with Trypanosoma vivax] Avaliação clínico and laboratorial de bovinos nelore infectados experimentalmente com Trypanosoma vivax. Pesquisa Veterinária Brasileira 21(4):157-161. Embrapa Gado de Corte, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. Avaliaram-se as alterações clínico-laboratoriais de seis bezerros Nelore, de ambos os sexos, inoculados experimentalmente com 107 organismos viáveis de Trypanosoma vivax, isolados de bovinos da região de Poconé, Estado de Mato Grosso. Os animais foram observados diariamente, durante 30 dias, quanto aos parâmetros de temperatura retal, volume globular (VG), parasitemia, produção de anticorpos, coloração de mucosas, comportamento e apetite. Deteminaram-se os níveis séricos de aspartato aminotransferase (AST), fosfatase alcalina (FA), gama glutamiltransferase (GGT), creatina kinase (CK), colesterol, uréia, creatinina, cálcio, fósforo e o perfil eletroforético das proteínas séricas aos 4, 8, 12, 16, 23 e 30 dias pós-inoculação (DPI). Durante os 6 meses seguintes, os animais foram observados semanalmente, avaliando-se a temperatura retal, o VG e a parasitemia. T. vivax foi evidenciado a partir do terceiro e quarto DPI em todos os bezerros e persistiu até o 30º DPI em cinco dos seis animais em estudo. Ocorreu um decréscimo significativo (p<0,05) do valor médio do VG (25%) aos dez DPI. Os animais não apresentaram qualquer alteração no quadro clínico, bem como na avaliação da bioquímica sérica durante o período experimental. A soroconversão ocorreu aos 6 e 8 DPI, permanecendo todos os animais soropositivos nos 30 dias experimentais. Bovinos nelores jovens, infectados experimentalmente com T. vivax, foram capazes de estabelecer um equilíbrio na relação hospedeiro-parasita.

Abstract in English:

ABSTRACT.- Lemos K.R. & Alessi A.C. 1999. [Glial fibrillary acidic protein (GFAP) immunoreactive astrocytes in the Central Nervous System of normal horses and horses with leukoencephalomalacia.] Astrócitos imunorreativos à proteína glial fibrilar ácida (GFAP) em sistema nervoso central de equinos normais e de equinos com leucoencefalomalácia. Pesquisa Veterinária Brasileira 19(3/4):104-108. Depto Patologia Veterinária, Faculdade de Ciências Agrárias e Veterinárias, Unesp, Rod. Carlos Tonanni Km 5, Jaboticabal, SP 14870-000, Brazil. The glial fibrillary acidic protein (GFAP), subunit of the intermediary filaments of the cellular cytoskeleton, exists in the cytoplasm of astrocytes. Immunohistochemistry utilizing primary antibodies anti-GFAP is generally chosen to identify astrocytes in the central nervous system (CNS), allowing also to verify their hypertrophy. Several studies show the distribution, morphology and cytoarchitecture of the astrocytes in several areas of the CNS of humans and laboratory animals. However, in domestic animals, especially in horses, little information is available. In the present study the density ánd morphology of GFAP-immunoreactive astrocytes in the white matter of the cerebral cortex of horses with leukoencephalomalacia (LEM) has been compared with such aspects in normal horses. In animals with LEM hypertrophic astrocytes in areas dose to the lesions were observed. There was enlargement of the perikarion, nucleus and the cytoplasmic extension. The astrocytes were reduced in number and the immunoreactivity was increased. In the normal animals constant distribution of immunoreactive cells characteristic of fibrous astrocytes was seen. Vascular changes in the animals with LEM, as for example degeneration of vascular endothelium, were also observed and could be correlated with the astrocytic alterations.

• Astrocytes GFAP horse leukoencephalomalacia Astrócito equino Ieucoencefalomalácia.

Abstract in Portuguese:

RESUMO.- Lemos K.R. & Alessi A.C. 1999. [Glial fibrillary acidic protein (GFAP) immunoreactive astrocytes in the Central Nervous System of normal horses and horses with leukoencephalomalacia.] Astrócitos imunorreativos à proteína glial fibrilar ácida (GFAP) em sistema nervoso central de equinos normais e de equinos com leucoencefalomalácia. Pesquisa Veterinária Brasileira 19(3/4):104-108. Depto Patologia Veterinária, Faculdade de Ciências Agrárias e Veterinárias, Unesp, Rod. Carlos Tonanni Km 5, Jaboticabal, SP 14870-000, Brazil. A proteína glial fibrilar ácida (GFAP), subunidade dos filamentos intermediários do citoesqueleto celular, está presente no citoplasma de astrócitos. Técnicas imunohistoquímicas com anticorpos primários anti-GFAP são geralmente empregadas para identificar astrócitos no sistema nervoso, permitindo verificar também sua hipertrofia. Vários estudos mostram a distribuição, á morfologia e a citoarquitetura de astrócitos em várias regiões do SNC do homem e de animais de laboratório. No entanto, em animais domésticos e, especialmente em equinos, poucas informações estão disponíveis. No presente trabalho, verificou-se a densidade e a morfologia de astrócitos imunorreativos à GFAP na substância branca da córtex cerebral de equinos com leucoencefalomalácia (LEM) comparando-se esses aspectos com o de equinos normais. Animais com LEM apresentaram hipertrofia de astrócitos em áreas próximas às lesões, representada pelo aumento do corpo celular, do núcleo e dos prolongamentos citoplasmáticos. O número de astrócitos apresentou-se reduzido e a imunorreatividade foi mais acentuada. Nos animais normais, verificou-se distribuição constante de astrócitos imunorreagentes com características de fibrosos. Alterações vasculares nos animais com LEM, como por exemplo degeneração de endotélio vascular, também foram observadas, podendo estar associadas às alterações astrocíticas.

Abstract in English:

An experimental study was conducted to evaluate the possibility of preventing Escherichia coli post-weaning diarrhea reducing the protein level and/or acidifying the diet. Sixteen SPF (Specific Pathogen Free) Landrace piglets, 36 days old (weaning day) were used in four treatments (T) as follow: TA) control ration with 20% crude protein (CP); TB) ration similar to TA with 2% citric acid; TC) ration with 16% CP plus lisine up to 0.9%; and TD) ration similar to TC plus 2% citric acid. Rations were formulated with corn, soybean meal, minerals and vitamins. The pigs received the experimental diets from Do (weaning day) to D10 (slaughter day). All piglets were inoculated with 20 ml of E. coli culture 0141 K85 with 109 CFU/ml on the day D3 and D4. The parameters analysed were, occurrence of diarrhea, faeces elimination and intestinal colonization with the E. coli sample inoculated, gastrointestinal content pH and small intestine histology. The addition of citric acid (TB) as well as CP reduction (TC) in the diet caused a benefic effect on the E. coli diarrhea. This effect was more evident with the CP reduction and addition of the acid simultaneously (TD). The diarrhea was prevented, probably, by the reduction of stomach and jejunum pH and by avoiding an exaggerated E. coli multiplication in the intestinal content.

• Pigs post-weaning diarrhea diet protein reduction diet acidification

Abstract in Portuguese:

Estudou-se experimentalmente a possibilidade de se prevenir a diarréia pós-desmama por Escherichia coli através da redução da proteína e/ou acidificação da dieta. Utilizaram-se 16 leitões Landrace SPF (Specific Pathogen Free) com 35 dias de idade (dia da desmama), distribuídos em quatro tratamentos (T): TA - ração controle com 20% de PB; T B - ração do TA com 2% de ácido cítrico; TC - ração com 16% PB mais lisina até nível de 0,9%; TD - ração do TC mais·2% do ácido cítrico. As rações foram formuladas com milho, farelo de soja, minerais e vitaminas. Os leitões receberam as dietas experimentais do D0 (dia da desmama) até D10 (dia do sacrifício). Todos os leitões foram inoculados com 20 ml de um cultivo de E. coli 0141 K85 contendo 109 UFC/ml, nos dias D3 e D4. Os parâmetros analisados foram: ocorrência de diarréia, eliminação nas fezes e colonização intestinal da amostra de E. coli inoculada, pH do conteúdo gastrointestinal e histologia do intestino delgado. Tanto a adição do ácido cítrico (TB) como a redução da PB (TC) na dieta tiveram efeito benéfico na ocorrência da diarréia por E. coli. Este efeito mostrou-se mais evidente com a redução da PB e adição do ácido simultaneamente (T D). A diarréia foi prevenida, possivelmente pela redução do pH do estômago e jejuno e por evitar a multiplicação exagerada de E. coli no conteúdo intestinal.

• Leitões diarréia pós-desmama redução de proteína acidificação

Abstract in English:

Thirty-two samples of buffalo milk from a group of 90 females from Pitangueiras County, State of São Paulo, were analyzed for pH, titratable acidity and total protein content. The results obtained showed positive· correlation between total protein content and titratable acidity r = 0. 76 (P < 0.01) but no relation to the real acidity or pH r = -0.34 (P > 0.05). On the other hand, of the 18 (56.25%) samples that had values of titratable acidity of over 18°D, 17 (53.13%) had normal values of pH from 6.45 to 6.80, permitting misinterpretation, if the titratable acidity of buffalo milk is evaluated using the parameters fixed for cow's milk. It was also seen that the data obtained contradict the alternative limits proposed for the modification of the acidity control system for this type of milk. For this reason, it is believed that the acidity determination must be based on the real acidity or pH, because parameters for titratable acidity based on the physico-chemical composition of buffalo milk have not been established.

• buffalo milk. titratable accidity pH total protein content regulatory patterns

Abstract in Portuguese:

Foram submetidas a determinação de acidez titulável, pH e teor de proteínas totais, 32 amostras de leite procedentes de 90 búfalas de uma propriedade situada no município de Pitangueiras, SP. Os resultados evidenciaram uma correlação positiva (r = 0,76; P < 0,01) entre o teor de proteínas totais e a acidez titulável sem influência evidente (r = 0,34; P>0,05) na acidez real ou pH. Por outro lado, das 18 (56,25%) amostras que apresentaram valores de acidez titulável superior a 18°D, 17 (53,13%) apresentaram valores de pH entre 6,45 e 6,80, evidenciando, portanto, a ocorrência de equívocos na interpretação, caso a acidez titulável do leite de búfala seja analisada com base nos padrões fixados para o leite de vaca. Verificou-se, ainda, que os dados obtidos contrariaram os limites alternativos propostos para a modificação do sistema de controle de acidez deste tipo de leite. Assim sendo, acredita-se que a análise de acidez deva ser efetuada com base na acidez real ou pH, enquanto não forem estabelecidos padrões de acidez titulável coerentes com a composição físico-química do leite de búfala.

• leite de búfala acidez titulável pH teor de proteínas totais padrões regulamentares

Abstract in English:

Fifty-five dairy cows from two herds located in the State of Rio de Janeiro, were evaluated monthly over a six month period for persistent lymphocytosis (PL) and for the presence of plasma antibody to the major glycoprotein (gp51) of bovine leukosis virus (BLV), as evidence of infection. Absolute lymphocyte counts were superimposed on Bendixen's hematological key for bovine leukosis. Animals were considered to have PL when, during the six month period, three absolute lymphocyte counts were above 7000 per mm3 of blood. In herd A, of 15 absolute lymphocyte counts from three antibody-negative cows, 13 (86.7%) were normal and two (13.3%) suspect. Counts from 17 antibody-positive cows from the sarne herd showed 29 (30.5%) normal, 36 (37.9%) suspect and 30 (31.6%) leukotic. In herd B, of 82 absolute lymphocyte counts from 14 antibody-negative cows, 45 (54.9%) were normal, 23 (28.1%) suspect and 14 (17.0%) leukotic. Counts from 21 antibody-positive cows showed that 53 (45.3%) were normal, 40 (34.2%) were suspect and 24 (20.5%) were leukotic. It is concluded that PL is an unreliable criterion to identify cattle infected with BLV in the subtropical environment, and must not be used in programs aimed at controlling or eradicating BLV from infected herds. However, the hematological keys will continue to be useful in the diagnosis of the clinical disease, that is, the bovine lymphosarcoma. The use of the immunodiffusion test to detect antibody to gp51 in the plasma or serum of infected cattle is a simple and sensitive tool in the control of BLV infection.

• Enzootic bovine leukosis virus persistent lymphocytosis antibodies glycoprotein

Abstract in Portuguese:

Cinqüenta e cinco vacas leiteiras de dois rebanhos localizados no Estado do Rio de Janeiro foram examinadas mensalmente, durante um período de seis meses. para determinar a linfocitose persistente (LP) e a presença, no plasma, de anticorpos contra a glicoproteina maior (gp51) do vírus da leucose bovina (VLB), como evidência da infecção. As contagens linfocitárias absolutas foram sobrepostas à chave hematológica de Bendixen para leucose bovina. Os animais foram considerados portadores de LP quando, durante o período de seis meses, três contagens linfocitárias absolutas estavam acima de 7000 por mm3 de sangue. No rebanho A, de 15 contagens linfocitárias absolutas de três vacas anticorpo-negativas, 13 (86,7%) eram normais e duas (13,3%) atingiram a faixa de suspeitas. De 95 contagens de 17 vacas anticorpo-positivas do mesmo rebanho, ,29 mostraram-se (30,5%) normais, 36 (37,9%) suspeitas e 30 (31,6%) leucóticas. No rebanho B, de 82 contagens linfocitárias absolutas de 14 vacas anticorpo-negativas; 45 (54,9%) eram normais, 23 (28,1 %) suspeitas e 14 (17,0%) leucóticas. As 117 contagens de 21 vacas anticorpo-positivas mostraram que 53 (45,3%) eram normais, 40 (34,2%) eram suspeitas e 24 (20,5%) eram leucóticas. Concluiu-se que a LP é um critério pouco confiável na detecção de bovinos infectados com o VLB num ambiente subtropical e não deve ser utilizada em programas que objetivam a controlar ou erradicar o VLB de rebanhos infectados. Porém, as chaves hematológicas continuarão sendo úteis no diagnóstico da doença clínica, o linfossarcoma bovino. Ficou também evidente que a prova de imunodifusão para detectar anticorpos contra gp51 no plasma ou soro de animais infectados é um método simples e sensível para o controle da leucose bovina.

• Leucose enzoótica bovina vírus linfocitose persistente anticorpos glicoproteina