PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

The objective of the present study was to detect the genetic diversity of Anaplasma marginale strains in naturally infected calves from a rural property located in the northeastern region of the state of Pará, Eastern Amazon, which has a history of mortality due to anaplasmosis. Fourteen calves positive for A. marginale were selected using a semi-nested polymerase chain reaction for the target msp1α gene, with asymptomatic (n=3) and symptomatic (n=11) infections. After sequencing the samples, two genotypes were verified in the E and C regions and the structures in tandem repeats were determined. Nine different strains were found: eight related to the E genotype (α-β-β-Γ = one animal, asymptomatic; 16-F-17-F-F = two animals, symptomatic; α-β-F-F-F-F = one animal, asymptomatic; 31-62-62-61 = one animal, symptomatic; τ-10-3 = three animals, two symptomatic and one asymptomatic; α-β-β-β = one animal, symptomatic; τ-22 -13-18 = two animals, both symptomatic; β-β-β-BRA1-31 = two animals, both symptomatic), and one related to genotype C (23-24-25-31-27-27 = one animal, asymptomatic). Genotype E was predominant in 92.86% of the samples (13/14), followed by genotype C (7.14%). This study made it possible to detect the genetic diversity of A. marginale in calves from the selected dairy farm, in addition to identifying the BRA1 sequence in the animals of the present study, which was recently diagnosed in Minas Gerais, demonstrating the dispersion of A. marginale strains in herds from different Brazilian states. Genetic diversity of A. marginale was observed in both symptomatic and asymptomatic calves. There were no significant differences when clinical signs were compared to the genotype verified in the infected animals. The prevalence of pathogenicity was not observed.

• Semi-nested PCR sequencing Anaplasma marginale msp1α BRA1 calves

Abstract in Portuguese:

O objetivo do presente trabalho foi detectar a diversidade genética de cepas de Anaplasma marginale em bezerros naturalmente infectados oriundos de uma propriedade rural localizada na região nordeste do estado do Pará, Amazônia Oriental, a qual apresentava histórico de mortalidade devido à anaplasmose. Foram selecionados 14 bezerros positivos para A. marginale pela técnica de semi-nested PCR (nPCR) para o alvo no gene msp1α, com infecção assintomática (n=3) e sintomáticos (n=11). Após o sequenciamento das amostras foram verificados dois genótipos nas regiões E e C, e determinadas as estruturas em tandem repeats. Nove diferentes estirpes foram encontradas, sendo oito relacionadas ao genótipo E (α-β-β-Γ = um animal, assintomático; 16-F-17-F-F = dois animais, sintomáticos; α-β-F-F-F-F = um animal, assintomático; 31-62-62-61 = um animal, sintomático; τ-10-3 = três animais, dois sintomáticos e um assintomático; α-β-β-β = um animal, sintomático; τ-22-13-18 = dois animais, sintomáticos; β-β-β-BRA1-31 = dois animais, sintomáticos) e uma relacionada ao genótipo C (23-24-25-31-27-27 = um animal, assintomático). O genótipo E foi predominante em 92,86% das amostras (13/14), seguido pelo genótipo C (7,14%). O estudo possibilitou a detecção da diversidade genética de A. marginale em bezerros dessa propriedade leiteira, além de identificar a sequência BRA1 nos animais do presente estudo, a qual foi diagnosticada recentemente em Minas Gerais, o que demonstra a dispersão das estirpes de A. marginale nos rebanhos de diferentes estados brasileiros. A diversidade genética de A. marginale foi observada tanto em bezerros sintomáticos quanto em assintomáticos e não houve diferença significativa quando se comparou os sinais clínicos ao genótipo verificado no animal infectado, não observando a prevalência de patogenicidade de estirpes.

• Semi-nested PCR sequenciamento Anaplasma marginale msp1α BRA1 bezerros

Abstract in English:

The present study aimed to analyze the casuistry of xenarthrans treated at the veterinary hospital of the Federal University of Pará to quantify the most important cases for clinical care and identify the main species, sex, age group, origin, and destination of the animals admitted. The data were collected from January 2013 to August 2020. The variables analyzed were the number of animals by species, sex, age group, weight (kilograms), form of admission (rescue or guardianship), person responsible for the admission of the animal, origin, circumstances of admission, length of stay (days), and evolution or destination of the animal (release, captivity, death, euthanasia, or escape). Fisher’s exact test was used to determine whether disease incidence was related to sex and whether animal mortality was influenced by sex and disease. The chi-square test was used to determine whether mortality differed by age group. Ninety-two (92) xenarthrans were included: 66 Bradypus variegatus, 10 Choloepus didactylus, 15 Tamandua tetradactyla, and one Dasypus novemcinctus. The animals were mainly from the municipality of Castanhal (70%) and were acquired through rescue (88%, B. variegatus; 80%, C. didactylus; 100%, T. tetradactyla), a significant proportion of which was per natural persons (88%, B. variegatus; 80%, C. didactylus; 100%, T. tetradactyla). The most important circumstances that led to the admission of the animals were as follows: animals for examination (42%, n=39), young animals at risk (9%, n=8), hit by a vehicle (8%, n=7), respiratory disease (7%, n=6), fall from a tree (5%, n=5), and electrotrauma (5%, n=5). Release was the most common destination of animals after hospitalization (59% B. variegatus, 50% C. didactylus, and 66% T. tetradactyla). The sex of the animals did not affect the incidence of disease or mortality in any species. In contrast, mortality of B. variegatus was significantly higher in animals admitted with disease (p=0.000). Mortality in cubs of this species was higher than that in juveniles and adults (p=0.003).

• Retrospective study amazon region wild fauna xenarthrans sloths anteater armadillo

Abstract in Portuguese:

O objetivo deste trabalho foi analisar a casuística dos Xenarthras atendidos no Hospital Veterinário da Universidade Federal do Pará (UFPA), a fim de quantificar as principais ocorrências para atendimento clínico, identificar as principais espécies, o sexo, a faixa etária, a procedência e destinação dos animais recebidos. Os dados foram obtidos entre janeiro de 2013 a agosto de 2020. As variáveis analisadas foram número de animais por espécie, sexo, faixa etária, peso (quilogramas), forma de recebimento (resgate, tutela), responsável pelo encaminhamento do animal ao hospital, procedência, circunstância de admissão, tempo de internação (dias) e evolução ou destinação do animal recebido (soltura, cativeiro, morte, eutanásia, fuga). O teste de Fisher foi utilizado para avaliar se a ocorrência de doenças estava relacionada ao sexo, bem como se a mortalidade animal foi influenciada pelo sexo e por enfermidades. O teste Qui-quadrado foi utilizado para avaliar se a mortalidade diferiu em relação à faixa etária. Um total de 92 animais Xenarthras foi admitido, composto por 66 Bradypus variegatus, 10 Choloepus didactylus, 15 Tamandua tetradactyla e um Dasypus novemcinctus. Os animais foram procedentes principalmente do município de Castanhal (70%), sendo o resgate a principal forma de aquisição dos animais (88%, B. Variegatus; 80%, C. Didactylus; 100%, T. tetradactyla), realizado em maior parte por pessoa física (88%, B. Variegatus; 80%, C. Didactylus; 100%, T. tetradactyla). O sexo dos animais não influenciou a ocorrência de doenças ou mortalidade em nenhuma espécie. Por outro lado, a mortalidade de B. variegatus foi significativamente maior naqueles animais admitidos com alguma enfermidade (p=0,000). A mortalidade em filhotes desta espécie foi maior quando comparada com as categorias jovem e adulta (p=0,003).

• Estudo retrospectivo região amazônica fauna silvestre Xenarthras bichos-preguiça tamanduá tatu

Abstract in English:

ABSTRACT.- Cepeda M.B., Cepeda P.B., Baêta B.A., Gaudêncio F.N., Magalhães-Matos P.C., Cordeiro M.D., Brito M.F. & Fonseca A.H. 2016. [Biochemical, anatomical and histopathological changes in the liver of Gallus gallus Linnaeus, 1758 experimentally infected by Borrelia anserine Sakharoff, 1891.] Alterações bioquímicas, anatômicas e histopatológicas em fígado de Gallus gallus Linnaeus, 1758, experimentalmente infectados por Borrelia anserina Sakharoff, 1891. Pesquisa Veterinária Brasileira 36(8):687-693. Departamento de Parasitologia Animal, Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, BR-465 Km 7, Seropédica, RJ 23890-000, Brazil. E-mail: adivaldofonseca@yahoo.com Spirochetosis avian is a septicemic disease of acute course and cosmopolitan can affect various avian species, caused by Borrelia anserina and transmitted by Argas miniatus. The experiment aimed to evaluate the biochemical, anatomical and histopathological changes in the liver of Gallus gallus caused by experimental infection with B. anserina. A total of 40 fowls of the species G. gallus were divided into four randomized groups of ten fowls each: G1 - inoculated with serum infected with B. anserina; G2 - inoculated with 0.9% saline; G3 - exposed to nymphs of 3rd instar of A. miniatus infected with B. anserina; G4 - exposed to ticks nymphs of 3rd instar of A. miniatus free of B. anserina. The fowls of Groups 1 and 3 expressed at 3 and 6 days post-inoculation (DAI) respectively , symptoms characteristic of the disease as lack of appetite , weight loss , drowsiness, greenish diarrhea, pale mucous membranes , ruffled feathers and hyperthermia. ALT of group 1 levels were significantly higher only at the 12º and 24º day after inoculation (DAI) compared with its control group (group 2), and in group 3 these levels remained high until the 20º DAI as compared with its control group (group 4). AST enzyme fluctuated little in the experimental groups, although elevations at 12ºDAI has been found in group 1 and 3. The liver of fowls in groups 1 and 3, presented at necropsy moderate hepatomegaly, congestion, irregular surface and red color to cyanotic. If found even small whitish spots on the surface. The histopathology revealed congestion, mononuclear inflammatory infiltrates, fibrinoid necrotic foci, dilatation of sinusoids, and vacuolation of hepatocytes. The Warthin-Starry staining revealed in the liver of fowls in groups 1 and 3 the presence of spirochetes compatible with B. anserina, often within blood vessels.

• Borrelia anserina spirochetosis tick borne diseases espiroquetose

Abstract in Portuguese:

RESUMO.- Cepeda M.B., Cepeda P.B., Baêta B.A., Gaudêncio F.N., Magalhães-Matos P.C., Cordeiro M.D., Brito M.F. & Fonseca A.H. 2016. [Biochemical, anatomical and histopathological changes in the liver of Gallus gallus Linnaeus, 1758 experimentally infected by Borrelia anserine Sakharoff, 1891.] Alterações bioquímicas, anatômicas e histopatológicas em fígado de Gallus gallus Linnaeus, 1758, experimentalmente infectados por Borrelia anserina Sakharoff, 1891. Pesquisa Veterinária Brasileira 36(8):687-693. Departamento de Parasitologia Animal, Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, BR-465 Km 7, Seropédica, RJ 23890-000, Brazil. E-mail: adivaldofonseca@yahoo.com A espiroquetose aviária é uma enfermidade septicêmica de curso agudo, cosmopolita, que acomete diversas espécies aviárias, causada por Borrelia anserina e transmitida pelo carrapato Argas miniatus. O experimento teve como objetivos avaliar as alterações bioquímicas e anátomo-histopatológicas no fígado de Gallus gallus, causadas pela infecção experimental por B. anserina. Quarenta aves da espécie G. gallus foram divididas em quatro grupos inteiramente casualizados com 10 animais cada: G1 - inoculado com soro infectado com B. anserina; G2 - inoculado com soro fisiológico a 0,9%; G3 - exposto a ninfas de terceiro ínstar de A. miniatus infectados por B. anserina; G4 - exposto a ninfas de terceiro ínstar de A. miniatus livres de B. anserina. As aves dos Grupos 1 e 3 manifestaram no 3º e 6º dias pós-inoculação (DPI) respectivamente, sintomatologia característica da doença como inapetência, perda de peso, sonolência, diarreia esverdeada, mucosas hipocoradas, penas arrepiadas e hipertermia. Os níveis de ALT do Grupo 1 mostraram-se significativamente mais elevados apenas no 12ºDPI e 24ºDPI em relação ao seu grupo controle (Grupo 2) e no Grupo 3 esses níveis se mantiveram elevados até o 20º DPI em comparação ao seu grupo controle (Grupo 4). Os níveis da enzima AST pouco oscilaram nos grupos experimentais, embora tenham sido encontradas elevações no 12ºDPI nos Grupos 1 e 3. Os fígados das aves dos Grupos 1 e 3 apresentaram à necropsia, moderada hepatomegalia, congestão, superfície irregular e coloração vermelha a cianótica; constataram-se ainda pequenos pontos esbranquiçados na superfície. A histopatologia do fígado revelou congestão, infiltrados inflamatórios mononucleares, focos de necrose fibrinoide, dilatação dos sinusoides e vacuolização de hepatócitos. A coloração de Warthin-Starry revelou, nos fígados das aves dos Grupos 1 e 3, a presença de espiroquetas compatíveis com B. anserina, frequentemente no interior de vasos sanguíneos.

Abstract in English:

ABSTRACT.- Silva J., Moraes C.M., Silva C.L., Sales G.A., Keid L.B., Matos P.C.M., Lara A.P.S.S. & Moraes C.C.G. 2016. Brucella abortus detected in cheese from the Amazon region: differentiation of a vaccine strain (B19) from the field strain in the states of Pará, Amapá and Rondônia, Brazil. Pesquisa Veterinária Brasileira 36(8):705-710. Laboratório de Zoonoses e Saúde Pública, Programa de Pós-Graduação em Saúde Animal na Amazônia, Universidade Federal do Pará, BR-316 Km 62, Saudade, Castanhal, PA 68743-050, Brazil. E-mail: ccmoraes@ufpa.br Brucellosis is an infectious-contagious disease responsible for significant economic losses to the meat and milk supply chain, because it causes reproductive disorders in animals and is a chronic anthropozoonosis. This study was designed to detect the DNA of Brucella spp. in cheese and to differentiate between a vaccine strain (B19) and the field strain. Sixty-six samples of different cheeses which are produced and marketed in three states of the Brazilian Amazon region (Amapá [5 samples], Pará [55 samples] and Rondônia [6 samples]) were evaluated. Thirty-nine of these samples were from cheeses made from cow’s milk, and 27 were from cheeses made from buffalo milk. Four of the 66 samples were from cheeses produced in milk processing plants regulated by the Federal Inspection Service (Serviço de Inspeção Federal); nine of the samples were from cheeses produced in processing plants regulated by the State Inspection Service (Serviço de Inspeção Estadual); five of the samples were from artisanal cheeses; and the remaining 48 samples were from informally produced cheese. DNA was obtained from the samples following a DNA extraction protocol, and PCR was conducted using primers B4 and B5 to detect Brucella spp. Primers eri1 and eri2 were used to differentiate the field strain from the B19 vaccine strain. The results showed that 21.21% (14/66) of the samples were positive for Brucella spp., of which 21.43% (3/14) were positive for the B. abortus field strain, and 7.14% (1/14) were identified as harboring vaccine strain B19. These results demonstrate that it is possible to identify Brucella spp. in cheese from the Amazon region using the PCR technique and to differentiate the B. abortus field strain from the B19 vaccine strain.

• Brucella abortus cheese vaccine (B19) queijos vacina B19

Abstract in Portuguese:

RESUMO.- Silva J., Moraes C.M., Silva C.L., Sales G.A., Keid L.B., Matos P.C.M., Lara A.P.S.S. & Moraes C.C.G. 2016. Brucella abortus detected in cheese from the Amazon region: differentiation of a vaccine strain (B19) from the field strain in the states of Pará, Amapá and Rondônia, Brazil. [Brucella abortus em queijos na região amazônica: diferenciação em cepa vacinal (B19) ou de infecção a campo nos estados do Pará, Amapá e Rondônia.] Pesquisa Veterinária Brasileira 36(8):705-710. Laboratório de Zoonoses e Saúde Pública, Programa de Pós-Graduação em Saúde Animal na Amazônia, Universidade Federal do Pará, BR-316 Km 62, Saudade, Castanhal, PA 68743-050, Brazil. E-mail: ccmoraes@ufpa.br A brucelose é uma enfermidade infecto-contagiosa que causa grandes perdas econômicas à cadeia produtiva da carne e do leite, como consequência dos distúrbios reprodutivos nos animais, além de ser uma antropozoonose crônica. O objetivo deste estudo foi detectar DNA de Brucella spp. e fazer a distinção da cepa vacinal (B19) da cepa de infecção de campo. Foram adquiridas 66 amostras de diferentes queijos produzidos e comercializados em três estados pertencentes à Amazônia brasileira: Amapá (05), Pará (55) e Rondônia (06), somando 39 amostras de queijo de vaca e 27 de búfala. Deste total quatro eram produzidas em estabelecimentos com fiscalização de Serviço de Inspeção Federal, nove em estabelecimentos com Serviço de Inspeção Estadual, cinco eram de produção artesanal e as demais 48 amostras eram provenientes de produção informal. O DNA das amostras teste foi obtido por um protocolo de extração e a reação em cadeia pela polimerase foi realizada utilizando os oligoiniciadores B4 e B5 para detectar Brucella spp. e, os oligoiniciadores eri1 e eri2 para diferenciar cepa de infecção a campo da cepa vacinal B19. Os resultados mostraram que 21,21% (14/66) das amostras foram positivas para Brucella spp., destas 21,43% (3/14) foram positivas para B. abortus cepa de campo e 7,14% (1/14) foi identificada como cepa vacinal B19. Concluiu-se que foi possível identificar pela técnica da PCR Brucella spp. em queijos na região amazônica, além de diferenciar as cepas em amostra de B. abortus de infecção a campo ou cepa vacinal B19.

Abstract in English:

ABSTRACT.- Leal D.C., Madruga C.R., Matos P.F., Souza B.M.P.S. & Franke C.R. 2011. Evaluation of PCR and multiplex PCR in relation to nested PCR for diagnosing Theileria equi. Pesquisa Veterinária Brasileira 31(7):575-578. Departamento de Produção Animal, Escola de Medicina Veterinária, Universidade Federal da Bahia, Av. Adhemar de Barros 500, Ondina, Salvador, BA 40110170, Brazil. E-mail: franke@ufba.br Conventional PCR (PCRTeq) for diagnosing Theileria equi and multiplex PCR (M/PCRTeq-Bc) for diagnosing T. equi and Babesia caballi were comparatively evaluated with nested PCR (N/PCR-Teq) for diagnosing equine piroplasmosis. In DNA sensitivity determinations, in multiple dilutions of equine blood that had tested positive for T. equi, PCR-Teq and N/PCR-Teq detected hemoparasite DNA in the larger dilutions (1:128), but did not differ significantly from the M/PCRTeq-Bc (1:64). In analyses on equine serum tested by ELISA, there was high agreement between this serological test and PCR-Teq (k = 0.780) and moderate agreement with N/PCR-Teq (k = 0.562) and M/PCRTeq-Bc (k = 0.488). PCR-Teq found a higher frequency of T. equi both in extensively and intensively reared horses, but this was not significant in relation to N/PCR-Teq (P>0.05), and both PCRs indicated that there was an endemic situation regarding T. equi in the population of horses of this sample. PCR-Teq was only significantly different from M/PCR-Teq-Bc (P<0.05). PCR-Teq presented high sensitivity and specificity, comparable to N/PCR-Teq, but with the advantage of higher speed in obtaining results and lower costs and risks of laboratory contamination. This accredits PCR-Teq for epidemiological studies and for determinations on affected horses.

• Theileria equi

Abstract in Portuguese:

RESUMO.- Leal D.C., Madruga C.R., Matos P.F., Souza B.M.P.S. & Franke C.R. 2011. Evaluation of PCR and multiplex PCR in relation to nested PCR for diagnosing Theileria equi. Pesquisa Veterinária Brasileira 31(7):575-578. Departamento de Produção Animal, Escola de Medicina Veterinária, Universidade Federal da Bahia, Av. Adhemar de Barros 500, Ondina, Salvador, BA 40110170, Brazil. E-mail: franke@ufba.br Uma PCR convencional (PCRTeq) para diagnóstico de Theileria equi e PCR multiplex (M/PCRTeq-Bc) para diagnóstico T. equi e Babesia caballi foram avaliadas comparativamente a nested PCR (N/PCR-Teq) no diagnóstico de piroplasmose equina. Na determinação da sensibilidade com DNA em diluições múltiplas de sangue de equino positivo para T. equi, as PCR-Teq e N/PCR-Teq detectaram DNA do hemoparasito nas diluições maiores (1:128), mas não diferiu significativamente da M/PCRTeq-Bc (1:64). Na análise com soros de equinos testados por ELISA houve uma concordância elevada entre esse teste sorológico e a PCR-Teq (k =0,780) e moderada com N/PCR-Teq (k = 0.562) e M/PCRTeq-Bc (k = 0.488). A PCR-Teq determinou freqüência maior de T. equi tanto nos equinos de criação extensiva como na intensiva, entretanto essa não foi significativa com relação N/PCR-Teq (P>0.05), e ambas PCRs indicaram que há uma situação endêmica para T. equi na população de equinos que constaram da amostragem. A PCR-Teq diferiu significativamente apenas para a M/PCRTeq-Bc (P<0.05). A PCR-Teq apresentou alta sensibilidade e especificidade comparável a N/PCR-Teq, mas com a vantagem de maior rapidez na obtenção dos resultados e menor custo e risco de contaminação no laboratório. Isso credencia a PCR-Teq para estudos epidemiológicos e para determinação de equinos portadores.

Abstract in English:

ABSTRACT. Riet-Correa F., Haraguchi M., Dantas A.F., Burakovas R.G., Yokosuka A., Mimaki Y., Medeiros R.M.T. & Matos P.F. 2009. Sheep poisoning by Panicum dichotomiflorum in northeastern Brazil. Pesquisa Veterinária Brasileira 29(1):94-98. Hospital Veterinário, Centro de Saúde e Tecnologia Rural, Universidades Federal de Campina Grande, Patos, PB 58700-000, Brazil. E-mail: franklin.riet@pq.cnpq.br Different species of Panicum, including P. dichotomiflorum, have been reported as a cause of photosensitization in sheep, horses, cattle and goats. An outbreak of hepatogenous photosensitization occurred in 3 flocks of hair sheep in the Brazilian semiarid region. Eighty one out of 365 sheep were affected and 39 died. The main affected animals were nursing lambs and sheep younger than one year old. Donkeys, goats and cattle grazing in the same pasture were not affected. Clinical signs were edema of the head, followed by dermatitis, mainly in the face, ears, and croup, ocular discharge, corneal opacity with blindness, and redness of the coronary band and hoof. At necropsy of one affected lamb the liver was yellowish. Upon histologic examination scattered necrotic hepatocytes were observed in the liver and focal areas of necrosis of myocytes appeared in the heart. Samples of P. dicotomiflorum were analyzed by TLC and those containing saponins were isolated by HPLC using RP-C18 column and eluted with a mixture of MeOH and H2O. The isolated compounds were submitted to 1H and 13C NMR spectroscopy. Reactions were positive to furostanol saponins with the same Rf of the standard protodioscin (0.21) and methylprotodioscin (0.32). The spectroscopic results indicated a mixture of (25R)- and (25S)-protodioscin isomers in a proportion of 3:1, and methylprotodioscin.

• Lithogenic steroidal saponins Panicum dichotomiflorum photosensitization protodioscin

Abstract in Portuguese:

ABSTRACT. Riet-Correa F., Haraguchi M., Dantas A.F., Burakovas R.G., Yokosuka A., Mimaki Y., Medeiros R.M.T. & Matos P.F. 2009. Sheep poisoning by Panicum dichotomiflorum in northeastern Brazil. Pesquisa Veterinária Brasileira 29(1):94-98. Hospital Veterinário, Centro de Saúde e Tecnologia Rural, Universidades Federal de Campina Grande, Patos, PB 58700-000, Brazil. E-mail: franklin.riet@pq.cnpq.br Different species of Panicum, including P. dichotomiflorum, have been reported as a cause of photosensitization in sheep, horses, cattle and goats. An outbreak of hepatogenous photosensitization occurred in 3 flocks of hair sheep in the Brazilian semiarid region. Eighty one out of 365 sheep were affected and 39 died. The main affected animals were nursing lambs and sheep younger than one year old. Donkeys, goats and cattle grazing in the same pasture were not affected. Clinical signs were edema of the head, followed by dermatitis, mainly in the face, ears, and croup, ocular discharge, corneal opacity with blindness, and redness of the coronary band and hoof. At necropsy of one affected lamb the liver was yellowish. Upon histologic examination scattered necrotic hepatocytes were observed in the liver and focal areas of necrosis of myocytes appeared in the heart. Samples of P. dicotomiflorum were analyzed by TLC and those containing saponins were isolated by HPLC using RP-C18 column and eluted with a mixture of MeOH and H2O. The isolated compounds were submitted to 1H and 13C NMR spectroscopy. Reactions were positive to furostanol saponins with the same Rf of the standard protodioscin (0.21) and methylprotodioscin (0.32). The spectroscopic results indicated a mixture of (25R)- and (25S)-protodioscin isomers in a proportion of 3:1, and methylprotodioscin.

Abstract in English:

Abstract.- Figueiredo-Fernandes A., Ferreira-Cardoso J.V., Garcia-Santos S., Monteiro S.M., Carrola J., Matos P. & Fontaínhas-Fernandes A. 2007. Histopathological changes in liver and gill epithelium of Nile tilapia, Oreochromis niloticus, exposed to waterborne copper. Pesquisa Veterinária Brasileira 27(3):103-109. University of Trás-os-Montes and Alto Douro, Center of Studies on Technological of Environmental and Life Sciences (CETAV), Apartado 1013, Vila Real 5001-801, Portugal. E-mail: fontain@utad.pt Nile tilapia, Oreochromis niloticus, of both sexes were reared in freshwater and exposed to 0.5, 1.0 and 2.5mg L-1 of waterborne copper for a period of 21 days. Liver and gill samples were collected after 21 days of exposure to copper and lesions were analyzed by light microscopy. The main histopathological changes observed in gills exposed to the highest concentration were edema, lifting of lamellar epithelia and an intense vasodilatation of the lamellar vascular axis. Although less frequent, lamellar fusion caused by the filamentar epithelium proliferation and some lamellar aneurisms were also found. The liver of control group exhibited a quite normal architecture, while the fish exposed to copper showed vacuolation and necrosis. These hepatic alterations were more evident in fish exposed to 1.0 and 2.5mg L-1 copper concentrations. The number of hepatocytes nucleus per mm2 of hepatic tissue decreased with the increase of copper concentration. In contrast, the hepatic somatic index was high in fish exposed at 2.5mg L-1 of copper. In short, this work advance new knowledge as influence of copper in the gill and liver histology of O. niloticus and demonstrated that their effects could be observed at different concentrations.

• Copper gill liver histopathological changes Oreochromis niloticus Teleostei

Abstract in Portuguese:

Abstract.- Figueiredo-Fernandes A., Ferreira-Cardoso J.V., Garcia-Santos S., Monteiro S.M., Carrola J., Matos P. & Fontaínhas-Fernandes A. 2007. Histopathological changes in liver and gill epithelium of Nile tilapia, Oreochromis niloticus, exposed to waterborne copper. Pesquisa Veterinária Brasileira 27(3):103-109. University of Trás-os-Montes and Alto Douro, Center of Studies on Technological of Environmental and Life Sciences (CETAV), Apartado 1013, Vila Real 5001-801, Portugal. E-mail: fontain@utad.pt Nile tilapia, Oreochromis niloticus, of both sexes were reared in freshwater and exposed to 0.5, 1.0 and 2.5mg L-1 of waterborne copper for a period of 21 days. Liver and gill samples were collected after 21 days of exposure to copper and lesions were analyzed by light microscopy. The main histopathological changes observed in gills exposed to the highest concentration were edema, lifting of lamellar epithelia and an intense vasodilatation of the lamellar vascular axis. Although less frequent, lamellar fusion caused by the filamentar epithelium proliferation and some lamellar aneurisms were also found. The liver of control group exhibited a quite normal architecture, while the fish exposed to copper showed vacuolation and necrosis. These hepatic alterations were more evident in fish exposed to 1.0 and 2.5mg L-1 copper concentrations. The number of hepatocytes nucleus per mm2 of hepatic tissue decreased with the increase of copper concentration. In contrast, the hepatic somatic index was high in fish exposed at 2.5mg L-1 of copper. In short, this work advance new knowledge as influence of copper in the gill and liver histology of O. niloticus and demonstrated that their effects could be observed at different concentrations.