PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Araújo V.R., Lima-Verde I.B., Name K.P.O., Báo S.N., Campelo C.C., Silva J.R.V., Rodrigues A.P.R. & Figueiredo J.R. 2010. Bone Morphogenetic Protein-6 (BMP-6) induces atresia in goat primordial follicles cultured in vitro. Pesquisa Veterinária Brasileira 30(9):770-776. Programa de Pós-Graduação em Ciências Veterinárias, Laboratório de Manipulação de Oócitos e Folículos Pré-Antrais, Universidade Estadual do Ceará, Av. Paranjana 1700, Campus do Itaperi, Fortaleza, CE 60740-000, Brazil. E-mail: val_exclusiva@yahoo.com.br This study investigated the effects of bone morphogenetic protein 6 (BMP-6) on in vitro primordial follicle development in goats. Samples of goat ovarian cortex were cultured in vitro for 1 or 7 days in Minimum Essential Medium (control medium) supplemented with different concentrations of BMP-6. Follicle survival, activation and growth were evaluated through histology and transmission electron microscopy (TEM). After 7 days of culture, histological analysis demonstrated that BMP-6 enhanced the percentages of atretic primordial follicles when compared to fresh control (day 0). Nevertheless, BMP-6 increased follicular and oocyte diameter during both culture periods. As the culture period progressed from day 1 to day 7, a significant increase in follicle diameter was observed with 1 or 50ng/ml BMP-6. However, on the contrary to that observed with the control medium TEM revealed that follicles cultured for up to 7 days with 1 or 50ng/ml BMP-6 had evident signs of atresia. In conclusion, this study demonstrated that BMP-6 negatively affects the survival and ultrastructure of goat primordial follicles.

• Bone Morphogenetic Protein-6 folliculogenesis Proteína Morfogenética Óssea-6 foliculogênese

Abstract in Portuguese:

RESUMO.- Araújo V.R., Lima-Verde I.B., Name K.P.O., Báo S.N., Campelo C.C., Silva J.R.V., Rodrigues A.P.R. & Figueiredo J.R. 2010. Bone Morphogenetic Protein-6 (BMP-6) induces atresia in goat primordial follicles cultured in vitro. [A Proteína Morfogenética Óssea-6 (BMP-6) induz a atresia em folículos primordiais caprinos cultivados in vitro.] Pesquisa Veterinária Brasileira 30(9):770-776. Programa de Pós-Graduação em Ciências Veterinárias, Laboratório de Manipulação de Oócitos e Folículos Pré-Antrais, Universidade Estadual do Ceará, Av. Paranjana 1700, Campus do Itaperi, Fortaleza, CE 60740-000, Brazil. E-mail: val_exclusiva@yahoo.com.br O presente estudo investigou os efeitos da proteína morfogenética óssea-6 (BMP-6) no desenvolvimento in vitro de folículos primordiais caprinos. Amostras de córtex ovariano de cabras foram cultivados por 1 ou 7 dias em Meio Essencial Mínimo (meio controle) suplementado com diferentes concentrações de BMP-6. As taxas de sobrevivência, ativação e crescimento foram avaliadas por histologia clássica e microscopia eletrônica de transmissão (MET). Após 7 dias de cultivo, a análise histológica demonstrou que a BMP-6 aumentou o percentual de folículos primordiais degenerados no dia 7 quando comparados ao controle fresco (D0). Além disso, houve um aumento significativo do diâmetro folicular e oocitário em ambos os períodos de cultivo em todos os tratamentos na presença de BMP-6. Com a progressão do cultivo do dia 1 para o dia 7, nos tratamentos com 1 ou 50ng/ml de BMP-6, foi observado um aumento significativo no diâmetro folicular. Entretanto, contrário ao observado no meio controle, a MET revelou que os folículos cultivados nesses tratamentos apresentavam sinais evidentes de atresia. Em conclusão, esse estudo demonstrou que a BMP-6 afeta negativamente a sobrevivência e a ultra-estrutura de folículos primordiais caprinos.

Abstract in English:

ABSTRACT.- Alves F.R., Emerson T.F., Ricardo R.G., Juliana C.D., Antônio A.N.M.J., Ambrósio C.E., Irina K. & Miglino M.A. 2010. Establishment of a protocol for obtention of neuronal stem cells lineages from the dog olfactory epithelium. Pesquisa Veterinária Brasileira 30(4):363-372. Departamento de Ciência Animal, Campus Universitário, Bairro Cibrazen, Bom Jesus, PI 64900-000, Brazil. E-mail: flavioribeiro@ufpi.edu.br A morphological and cell culture study from nasal mucosa of dogs was performed in order to establish a protocol to obtain a cell population committed to neuronal lineage, as a proposal for the treatment of traumatic and degenerative lesions in these animals, so that in the future these results could be applied to the human species. Twelve mongrel dogs of 60-day aged pregnancy were collected from urban pound dogs in São Paulo. Tissue from cribriform ethmoidal lamina of the fetuses was collected at necropsy under sterile conditions around 1h to 2h postmortem by uterine sections and sections from the fetal regions described above. Isolated cells of this tissue were added in DMEM/F-12 medium under standard conditions of incubation (5% CO2, >37°C). Cell culture based on isolated cells from biopsies of the olfactory epithelium showed rapid growth when cultured for 24 hours, showing phase-bright sphere cells found floating around the fragments, attached on culture flasks. After 20 days, a specific type of cells, predominantly ellipsoids or fusiform cells was characterized in vitro. The indirect immunofluorescence examination showed cells expressing markers of neuronal precursors (GFAP, neurofilament, oligodendrocyte, and III â-tubulin). The cell proliferation index showed Ki67 immunostaining with a trend to label cell groups throughout the apical region, while PCNA immunostaining label predominantly cell groups lying above the basal lamina. The transmission electron microscopy from the olfactory epithelium of dogs revealed cells with electron-dense cytoplasm and preserving the same distribution as those of positive cell staining for PCNA. Metabolic activity was confirmed by presence of euchromatin in the greatest part of cells. All these aspects give subsidies to support the hypothesis about resident progenitor cells among the basal cells of the olfactory epithelium, committed to renewal of these cell populations, especially neurons.

• Olfactory epithelium cell culture stem cell epitélio olfatório cultivo de células células-tronco

Abstract in Portuguese:

RESUMO.- Alves F.R., Emerson T.F., Ricardo R.G., Juliana C.D., Antônio A.N.M.J., Ambrósio C.E., Irina K. & Miglino M.A. 2010. Establishment of a protocol for obtention of neuronal stem cells lineages from the dog olfactory epithelium. Pesquisa Veterinária Brasileira 30(4):363-372. Departamento de Ciência Animal, Campus Universitário, Bairro Cibrazen, Bom Jesus, PI 64900-000, Brazil. E-mail: flavioribeiro@ufpi.edu.br Foi realizado um estudo morfológico e por cultivo celular a partir de células provenientes da mucosa olfatória de cães, como forma de estabelecer um protocolo de cultivo, como uma proposta para o tratamento de lesões traumáticas e nervosas degenerativas nestes animais e futuramente, para que tais resultados possam ser aplicados a espécie humana. Foram utilizados doze cães sem raça definida, a termo, oriundos de castrações do Centro de Controle de Zoonoses de São Paulo. O tecido da lâmina cribiforme do etmóide dos fetos foi coletado sob necropsia, em condições estéreis, 1 a 2 horas post mortem, por meio de incisão uterina e acesso da região fetal supracitada. Depois as células isoladas desse tecido foram adicionadas em médio DMEM/F-12 sob condições padrão (5% CO2, >37°C). As células obtidas a partir de biópsias do epitélio olfatório de cães apresentaram rápido crescimento após 24 horas de cultivo, demonstrando morfologia esférica, sendo encontradas flutuando ao redor do fragmento aderido à garrafa de cultura. Após 20 dias, foram verificados tipos celulares específicos, predominantemente elipsóides ou fusiformes, foram observadas in vitro. Sob avaliação por imunofluorescência indireta observaram-se células com expressão positiva para marcadores de precursores neuronais (GFAP, Neurofilamentos, oligodendrócitos e â-tubulina III). O índice de proliferação celular mostrou-se positivo para Ki67 com uma tendência de marcação de grupos celulares ao longo da região apical, enquanto a imunomarcação para PCNA mostrou-se predominantemente em grupos celulares residentes sobre a lâmina basal. A microscopia eletrônica de transmissão do epitélio olfatório de cães revelou células com citoplasma eletrodenso e mesma distribuição das células marcadas positivamente para PCNA. A atividade metabólica foi confirmada pela presença de eucromatina em muitas regiões celulares. Todos estes aspectos sustentam a hipotese sobre a presença de células progenitoras residentes entre as células basais do epitélio olfatório comprometidas com a renovação desse epitélio, particularmente a população de neurônios.

Abstract in English:

ABSTRACT.- Cardinal S.G., Aniz A.C., Santos B.S., Carvalho N.M. & Lemos R.A.A. 2010. [Perinatal lesions caused in lambs by feeding Tetrapterys multiglandulosa (Malpighiaceae) to sheep at different stages of their pregnancy.] Lesões perinatais em ovinos causadas pela ingestão de Tetrapterys multiglandulosa (Malpighiaceae) em diferentes estágios de gestação. Pesquisa Veterinária Brasileira 30(1):73-78. Departamento de Medicina Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Federal de Mato Grosso do Sul, Av. Filinto Müller s/n, Cidade Universitária, Campo Grande, MS 79070-900, Brazil. E-mail: eqrural@nin.ufms.br In order to verify the effects of non-lethal doses of Tetrapterys multiglandulosa on ovine fetuses, experimental poisoning in sheep at different days of pregnancy was performed. Green leaves of shooting plants were administered to 9 pregnant ewes divided into three experimental groups. Sheep from Group 1 received four doses of 5g/kg of fresh leaves for 4 days; those from Group 2 received 10g/kg for 2 days; Group 3 sheep received a dose of 20g/kg for one day, and sheep from Group 4 did not receive the plant and served as negative controls. To check the plant toxicity, a male sheep (positive control) received 5g/kg until the onset of clinical signs. No signs of poisoning were observed in pregnant ewes; three lambs died 1-5 days after birth, and a fourth lamb died within 3 months after have been born. The positive control died after 38 days of daily consumption of the plant, presenting tachypnea, tachycardia, drowsiness, incoordination, weakness and sudden death. All four dead lambs and the positive control sheep were necropsied. The gross lesions were whitish areas in the myocardium, increased lobular pattern of the liver, dark red lungs, metabolization of pericardial fat, and ruminal distention with free gas. Histological findings were cardiac fibrosis, cardiomyocyte necrosis, pulmonary congestion and edema, and spongy degeneration in subcortical cerebral white matter. Tetrapterys multiglandulosa resulted toxic for ovine fetuses at doses that were not suffient to induce clinical signs in the pregnant ewes, demonstrating that the poisoning may be a cause of death of lambs soon after birth, even several months after the ingestion of the plant has been discontinued.

• Poisonous plants Tetrapterys multiglandulosa Malpighiaceae planta poisoning

Abstract in Portuguese:

RESUMO.- Cardinal S.G., Aniz A.C., Santos B.S., Carvalho N.M. & Lemos R.A.A. 2010. [Perinatal lesions caused in lambs by feeding Tetrapterys multiglandulosa (Malpighiaceae) to sheep at different stages of their pregnancy.] Lesões perinatais em ovinos causadas pela ingestão de Tetrapterys multiglandulosa (Malpighiaceae) em diferentes estágios de gestação. Pesquisa Veterinária Brasileira 30(1):73-78. Departamento de Medicina Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Federal de Mato Grosso do Sul, Av. Filinto Müller s/n, Cidade Universitária, Campo Grande, MS 79070-900, Brazil. E-mail: eqrural@nin.ufms.br Foi realizada a intoxicação experimental em ovelhas em diferentes dias de gestação, 12 ovelhas foram dividas em quatro grupos. As ovelhas do Grupo 1 consumiram quatro doses de 5g/kg de folhas verdes durante quatro dias; ovinos do Grupo 2 consumiram duas doses de 10g/kg durante dois dias, o Grupo 3 consumiu uma dose de 20g/kg em um único dia e os ovinos do Grupo 4 não consumiram a planta (controle-negativo). Para testar a toxicidade da planta um ovino macho consumiu 5g/kg até manifestação dos sintomas. Não foram observados sinais clínicos da intoxicação nas ovelhas, mas três cordeiros tiveram morte perinatal, e um cordeiro morreu com três meses de idade; o ovino controle-positivo morreu após 38 dias de consumo diário da planta e os sinais clínicos foram taquipnéia, taquicardia, sonolência, incoordenação e fraqueza. Todos os quatro cordeiros e o ovino controle-positivo foram necropsiados e os achados de necropsia foram áreas esbranquiçadas no miocárdio, evidenciação do padrão lobular do fígado, pulmão vermelho enegrecido e rúmen acentuadamente distendido com presença de gases livres. Os achados histopatológicos foram fibrose cardíaca, necrose dos cardiomiócitos, congestão e edema pulmonar, congestão hepática centrolobular e degeneração esponjosa na região subcortical do encéfalo. Tetrapterys multiglandulosa demonstrou ser tóxica para os fetos ovinos em dosagens que não foram suficientes para induzir sinais clínicos nas ovelhas prenhes, e que a intoxicação pode ocorrer com morte de cordeiros logo após o parto, mesmo após meses de ter cessada a ingestão da planta.

Abstract in English:

ABSTRACT.- Srivastav A.K., Singh S., Mishra D. & Srivastav S.K. 2009. Ultimobranchial gland of freshwater catfish, Heteropneustes fossilis, in response to calcitonin administration. Pesquisa Veterinária Brasileira 29(12):963-968. Department of Zoology, DDU Gorakhpur University, Gorakhpur 273 009, India. E-mail: ajaiksrivastav@hotmail.com The absence o!!f a hypocalcemic effect of calcitonin (CT) in fishes has been suggested due to exceedingly high plasma levels of CT; the fish may be saturated with respect of circulating CT and therefore unable to respond to exogenously administered CT. Earlier it has been suggested that a hypocalcemic action of injected CT may be obscured by changes in the release of endogenous CT and other calcium regulating hormones. In this study we have used artificial freshwater, calcium-deficient freshwater and calcium-rich freshwater and injected the fish with CT. The aim behind selecting these media were (i) in calcium-deficient medium there would be reduced circulating levels of CT, (ii) in calcium-rich medium there would be diminished secretion of prolactin (this hormone is hypercalcemic in fish), and (iii) by keeping the fish in calcium-rich medium we can test the antihypercalcemic action of CT. Moreover, the present study would reveal the changes in the ultimobranchial gland (UBG) after keeping the fish in all the above three media and/or injecting the fish with CT. Freshwater catfish, Heteropneustes fossilis, were administered intraperitoneally daily with vehicle or 0.5 U/100g body wt of salmon calcitonin (CT) and kept in artificial freshwater, calcium-rich freshwater and calcium-deficient freshwater for 10 days. Blood samples were collected on 1, 3, 5, and 10 days following the treatment and analyzed for serum calcium levels. The ultimobranchial gland (UBG) was also fixed for histological studies on these intervals. In artificial freshwater there was no change in the serum calcium levels of calcitonin-injected fish. The ultimobranchial gland of calcitonin-injected fish exhibited a progressive decrease in the nuclear volume from day 5 onwards. On day 10 vacuolization in the gland was also noticed. In vehicle-injected fish (control) kept in calcium-rich freshwater hypercalcemia has been noticed which persists till the end of the experiment. In calcitonin-treated fish maintained in calcium-rich freshwater there is no change in serum calcium level as compared to vehicle-injected fish. In vehicle-injected fish the UBG depicts decreased staining response and increased nuclear volume at day 5. On day 10 the nuclear volume is further increased and few degenerating cells have been noticed. Calcitonin fails to induce any histological change in the UBG as compared to control. In vehicle-injected fish kept in calcium-deficient freshwater the serum calcium levels decrease from day 1 to day 3. The levels exhibit hypercalcemia on day 10. CT treatment to the fish kept in calcium-deficient freshwater evokes a decrease in the calcium levels on day 1 and day 3. A significant hypercalcemia has been noticed on day 5 and day 10. In vehicle-injected fish kept in calcium-deficient freshwater the UBG reveals a decreased staining response on day 10. In CT-injected fish maintained in calcium-deficient freshwater the UBG depicts an increased nuclear volume and few exhausted cells on day 10. It can be concluded that CT can provoke hypocalcemia only when the fish is kept in medium which reduces the circulating levels of this hormone. The UBG of the fish kept in different calcemic media responded in a manner to indicate that it produces hypocalcemic factor - CT.

• Calcitonin Heteropneustes fossilis

Abstract in Portuguese:

ABSTRACT.- Srivastav A.K., Singh S., Mishra D. & Srivastav S.K. 2009. Ultimobranchial gland of freshwater catfish, Heteropneustes fossilis, in response to calcitonin administration. Pesquisa Veterinária Brasileira 29(12):963-968. Department of Zoology, DDU Gorakhpur University, Gorakhpur 273 009, India. E-mail: ajaiksrivastav@hotmail.com The absence o!!f a hypocalcemic effect of calcitonin (CT) in fishes has been suggested due to exceedingly high plasma levels of CT; the fish may be saturated with respect of circulating CT and therefore unable to respond to exogenously administered CT. Earlier it has been suggested that a hypocalcemic action of injected CT may be obscured by changes in the release of endogenous CT and other calcium regulating hormones. In this study we have used artificial freshwater, calcium-deficient freshwater and calcium-rich freshwater and injected the fish with CT. The aim behind selecting these media were (i) in calcium-deficient medium there would be reduced circulating levels of CT, (ii) in calcium-rich medium there would be diminished secretion of prolactin (this hormone is hypercalcemic in fish), and (iii) by keeping the fish in calcium-rich medium we can test the antihypercalcemic action of CT. Moreover, the present study would reveal the changes in the ultimobranchial gland (UBG) after keeping the fish in all the above three media and/or injecting the fish with CT. Freshwater catfish, Heteropneustes fossilis, were administered intraperitoneally daily with vehicle or 0.5 U/100g body wt of salmon calcitonin (CT) and kept in artificial freshwater, calcium-rich freshwater and calcium-deficient freshwater for 10 days. Blood samples were collected on 1, 3, 5, and 10 days following the treatment and analyzed for serum calcium levels. The ultimobranchial gland (UBG) was also fixed for histological studies on these intervals. In artificial freshwater there was no change in the serum calcium levels of calcitonin-injected fish. The ultimobranchial gland of calcitonin-injected fish exhibited a progressive decrease in the nuclear volume from day 5 onwards. On day 10 vacuolization in the gland was also noticed. In vehicle-injected fish (control) kept in calcium-rich freshwater hypercalcemia has been noticed which persists till the end of the experiment. In calcitonin-treated fish maintained in calcium-rich freshwater there is no change in serum calcium level as compared to vehicle-injected fish. In vehicle-injected fish the UBG depicts decreased staining response and increased nuclear volume at day 5. On day 10 the nuclear volume is further increased and few degenerating cells have been noticed. Calcitonin fails to induce any histological change in the UBG as compared to control. In vehicle-injected fish kept in calcium-deficient freshwater the serum calcium levels decrease from day 1 to day 3. The levels exhibit hypercalcemia on day 10. CT treatment to the fish kept in calcium-deficient freshwater evokes a decrease in the calcium levels on day 1 and day 3. A significant hypercalcemia has been noticed on day 5 and day 10. In vehicle-injected fish kept in calcium-deficient freshwater the UBG reveals a decreased staining response on day 10. In CT-injected fish maintained in calcium-deficient freshwater the UBG depicts an increased nuclear volume and few exhausted cells on day 10. It can be concluded that CT can provoke hypocalcemia only when the fish is kept in medium which reduces the circulating levels of this hormone. The UBG of the fish kept in different calcemic media responded in a manner to indicate that it produces hypocalcemic factor - CT.

Abstract in English:

RESUMO.- Monteiro C.M.R., Perri S.H.V., Roberto Gameiro de Carvalho R.G. & Koivisto M.B. 2009. [Histology and morphometry in uterus horns of nulliparous bitches, multiparous ones and bitches treated with contraceptives.] Histologia e morfometria em cornos uterinos de cadelas nulíparas, multíparas e tratadas com contraceptivos. Pesquisa Veterinária Brasileira 29(10):847-851. Departamento de Clínica, Cirurgia e Reprodução Animal, Curso de Medicina Veterinária, Faculdade de Odontologia, Universidade Estadual Paulista, Rua Clóvis Pestana 173, Araçatuba, SP 16065-090, Brazil. E-mail: monteiro@fmva.unesp.br Dados histológicos e morfométricos foram obtidos de útero de cadelas nulíparas (n=6), multíparas (n=6) e de cadelas tratadas com contraceptivo (n=6). Para esse fim foram usadas seis amostras de cornos uterinos, em corte médio, para cada grupo. As mensurações das espessuras da parede uterina, endométrio total, miométrio total, miométrio interno, miométrio externo, estrato vascular e diâmetro das glândulas endometriais normais não foram estatisticamente significantes entre as cadelas multíparas e tratadas, com exceção para a altura do epitélio de glândulas normais. As mensurações das espessuras da parede uterina, endométrio total, miométrio total, miométrio interno, diâmetro das glândulas normais e altura do epitélio glandular foram significantes, comparando cadelas nulíparas com as multíparas e/ou tratadas, com exceção para as espessuras do miométrio externo e estrato vascular. Diante dos resultados obtidos, pode-se concluir que os dados numéricos confirmam que (1) o uso de anticoncepcionais e sucessivas gestações afetam a estrutura uterina em seu total; (2) o miométrio externo e estrato vascular foram as regiões que menos sofreram alterações comparando-se os 3 grupos; (3) as variações morfológicas ocorreram com a mesma intensidade no endométrio e miométrio totais para as cadelas tratadas e multíparas, e (4) a presença de glândulas endometriais dilatadas foi o diferencial encontrado nas cadelas tratadas, pois as mesmas estavam ausentes nas multíparas e nulíparas.

• Morphometry uterine horns endometrium myometrium bitches

Abstract in Portuguese:

ABSTRACT.- Monteiro C.M.R., Perri S.H.V., Roberto Gameiro de Carvalho R.G. & Koivisto M.B. 2009. [Histology and morphometry in uterus horns of nulliparous bitches, multiparous ones and bitches treated with contraceptives.] Histologia e morfometria em cornos uterinos de cadelas nulíparas, multíparas e tratadas com contraceptivos. Pesquisa Veterinária Brasileira 29(10):847-851. Departamento de Clínica, Cirurgia e Reprodução Animal, Curso de Medicina Veterinária, Faculdade de Odontologia, Universidade Estadual Paulista, Rua Clóvis Pestana 173, Araçatuba, SP 16065-090, Brazil. E-mail: monteiro@fmva.unesp.br Histological and morphometric data were obtained from uterus of nulliparous bitches (n=6), multiparous ones (n=6), and bitches treated with contraceptive (n=6). Six samples of uterine horns, cut in average for each group. Measurements of the thickness of the uterine wall, total endometrium, total myometrium, internal myometrium, external myometrium, vascular layer and diameter of normal endometrial glands were not statistically significant between the treated and multiparous bitches, except for the height of the epithelium of normal glands. Measurements of thickness of the uterine wall, total endometrium, total myometrium, myometrium internal diameter and height of the glands of normal glandular epithelium were significant, comparing nulliparous with multiparous bitches and/or treated, except for thickness of the myometrium and vascular outer layer. It can be concluded that (1) the use of contraceptives and successive pregnancies affected uterine structures in its full, (2) the vascular layer and external myometrium were the regions with less variation, (3) morphological changes occurred with the same intensity in the total endometrium and myometrium of treated and multiparous bitches; and (4) the presence of dilated endometrial glands was the difference found in the treated bitches, because they were absent in nulliparous and multiparous ones.

Abstract in English:

ABSTRACT.- Buim M.R., Mettifogo E., Timenetsky J., Kleven S. & Ferreira A.J.P. 2009. Epidemiological survey on Mycoplasma gallisepticum and M. synoviae by multiplex PCR in commercial poultry. Pesquisa Veterinária Brasileira 29(7):552-556. Departamento de Patologia, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo, SP 05508-900, Brazil. E-mail: ajpferr@usp.br Mycoplasmas are important avian pathogens, which cause respiratory and joint diseases that result in large economic losses in Brazilian and world-wide poultry industry. This investigation regarding the main species of mycoplasmas, Mycoplasma gallisepticum (MG) and M. synoviae (MS), responsible for the above mentioned conditions, was carried out through PCR Multiplex analysis. One thousand and forty-six (1,046) samples of tracheal swabs and piped embryos were collected from 33 farms with laying hens, breeders, broilers or hatchery, located in the Brazilian states of São Paulo, Paraná and Pernambuco, where respiratory problems or drops in egg production had occurred. The MG and MS prevalence on the farms was 72.7%. These results indicated (1) high dissemination of mycoplasmas in the evaluated farms, with predominance of MS, either as single infectious agent or associated with other mycoplasmas in 20 farms (60.6%), and (2) an increase of MS and decrease of MG infection in Brazilian commercial poultry.

• Mycoplasma gallisepticum M. synoviae PCR epidemiology chicken

Abstract in Portuguese:

ABSTRACT.- Buim M.R., Mettifogo E., Timenetsky J., Kleven S. & Ferreira A.J.P. 2009. Epidemiological survey on Mycoplasma gallisepticum and M. synoviae by multiplex PCR in commercial poultry. Pesquisa Veterinária Brasileira 29(7):552-556. Departamento de Patologia, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo, SP 05508-900, Brazil. E-mail: ajpferr@usp.br Mycoplasmas are important avian pathogens, which cause respiratory and joint diseases that result in large economic losses in Brazilian and world-wide poultry industry. This investigation regarding the main species of mycoplasmas, Mycoplasma gallisepticum (MG) and M. synoviae (MS), responsible for the above mentioned conditions, was carried out through PCR Multiplex analysis. One thousand and forty-six (1,046) samples of tracheal swabs and piped embryos were collected from 33 farms with laying hens, breeders, broilers or hatchery, located in the Brazilian states of São Paulo, Paraná and Pernambuco, where respiratory problems or drops in egg production had occurred. The MG and MS prevalence on the farms was 72.7%. These results indicated (1) high dissemination of mycoplasmas in the evaluated farms, with predominance of MS, either as single infectious agent or associated with other mycoplasmas in 20 farms (60.6%), and (2) an increase of MS and decrease of MG infection in Brazilian commercial poultry.

Abstract in English:

ABSTRACT.- Rangel P. & Marin J.M. 2009. Analysis of Escherichia coli isolated from bovine mastitic milk. Pesquisa Veterinária Brasileira 29(5):363-368. Departamento de Morfologia, Estomatologia e Fisiologia, Faculdade de Odontologia de Ribeirão Preto, Universidade de São Paulo, Avenida do Café s/n, Campus USP, Ribeirão Preto, SP 14040-904, Brazil. E-mail: jmmarin@forp.usp.br Mastitis has been recognized for some time as the most costly disease in dairy herds. From February to November 2004, 670 samples of bovine mastitic milk from which 231 Escherichia coli strains were isolated, were collected from two Brazilian states. The strains were screened for the presence of Shiga toxin-producing (stx 1 and stx 2) and intimin (eae) genes. Twenty (8.6%) strains were detected by PCR to harbor the Shiga toxin genes (8 the stx 1 gene, 12 the stx 2 gene and none both of them). Two (0.8%) of the Escherichia coli strains studied were eae positive non Shiga toxin-producing. The strains were also examined for resistance to 12 antimicrobial agents. The predominantly observed resistance was to tetracycline (92.2%), streptomycin (90.4%), nalidixic acid (88.3%), amikacin (86.5%) and cephalothin (84.8%). Multidrug resistance was found among 152 isolates (65.8%).

• Escherichia coli mastitis antimicrobial agents multidrug resistance eae gene mastite agentes antimicrobianos resistência a antimicrobianos gene eae.

Abstract in Portuguese:

ABSTRACT.- Rangel P. & Marin J.M. 2009. Analysis of Escherichia coli isolated from bovine mastitic milk. Pesquisa Veterinária Brasileira 29(5):363-368. Departamento de Morfologia, Estomatologia e Fisiologia, Faculdade de Odontologia de Ribeirão Preto, Universidade de São Paulo, Avenida do Café s/n, Campus USP, Ribeirão Preto, SP 14040-904, Brazil. E-mail: jmmarin@forp.usp.br Mastitis has been recognized for some time as the most costly disease in dairy herds. From February to November 2004, 670 samples of bovine mastitic milk from which 231 Escherichia coli strains were isolated, were collected from two Brazilian states. The strains were screened for the presence of Shiga toxin-producing (stx 1 and stx 2) and intimin (eae) genes. Twenty (8.6%) strains were detected by PCR to harbor the Shiga toxin genes (8 the stx 1 gene, 12 the stx 2 gene and none both of them). Two (0.8%) of the Escherichia coli strains studied were eae positive non Shiga toxin-producing. The strains were also examined for resistance to 12 antimicrobial agents. The predominantly observed resistance was to tetracycline (92.2%), streptomycin (90.4%), nalidixic acid (88.3%), amikacin (86.5%) and cephalothin (84.8%). Multidrug resistance was found among 152 isolates (65.8%).

Abstract in English:

BSTRACT.- Reis A.B., Sant’Ana D.M.G., Azevedo J.F., Merlini L.S. & Araújo E.J.A. 2009. [The influence of the aquatic environment in tanks sequetially interconnected with PVC pipes on the gill epithelium and lamellas of tilapia (Oreochromis niloticus).] Alterações do epitélio branquial e das lamelas de tilápias (Oreochromis niloticus) causadas por mudanças do ambiente aquático em tanques de cultivo intensivo. Pesquisa Veterinária Brasileira 29(4):303-311. Laboratório de Neurogastroenterologia Experimental, Universidade Paranaense, Praça Mascarenhas de Moraes 4286, Umuarama, PR 87502-210, Brazil. E-mail: eduardoaraujo@unipar.br The behavior of the gill epithelium of tilapias cultured in tanks at different altitudes and interconnected with PVC pipes was analyzed. Gill filaments of four specimens from four tanks (T1, T2, T3 e T4) sequentially interconnected were submitted to histological routine to obtain 5-mm-thick cuts that were stained with HE or submitted to histochemistry reactions PAS + diastase solution or Alcian Blue pH 2.5 or Alcian Blue pH 1.0. Considering the intermediary, apical and basal regions of the filaments, the lamellar area was measured and the amount of mucous cells was counted. It was verified that oxygen, pH, and temperature decreased progressively as the water flew from one tank to another. Thus, an increase was realized of the amount of mucous cells and the lamellar area in T2, as well as a progressive decrease of these measures on the tanks which received water from T2. Moreover, detachment of the gill epithelium, cellular hyperplasia in the interlamellar space, and telangectasias were observed in the fishes from T2, T3 e T4. It was concluded that the aquatic environment in tanks sequentially interconnected with PVC pipes suffers alterations from one tank to another, as that physico-chemical fluctuations reflect on the behavior of the gill epithelium through variations of the lamellar area and the amount of mucous cells.

• Nile tilapia intensive culture system gill; morphometry mucous cells secundary lamellar cells

Abstract in Portuguese:

BSTRACT.- Reis A.B., Sant’Ana D.M.G., Azevedo J.F., Merlini L.S. & Araújo E.J.A. 2009. [The influence of the aquatic environment in tanks sequetially interconnected with PVC pipes on the gill epithelium and lamellas of tilapia (Oreochromis niloticus).] Alterações do epitélio branquial e das lamelas de tilápias (Oreochromis niloticus) causadas por mudanças do ambiente aquático em tanques de cultivo intensivo. Pesquisa Veterinária Brasileira 29(4):303-311. Laboratório de Neurogastroenterologia Experimental, Universidade Paranaense, Praça Mascarenhas de Moraes 4286, Umuarama, PR 87502-210, Brazil. E-mail: eduardoaraujo@unipar.br The behavior of the gill epithelium of tilapias cultured in tanks at different altitudes and interconnected with PVC pipes was analyzed. Gill filaments of four specimens from four tanks (T1, T2, T3 e T4) sequentially interconnected were submitted to histological routine to obtain 5-mm-thick cuts that were stained with HE or submitted to histochemistry reactions PAS + diastase solution or Alcian Blue pH 2.5 or Alcian Blue pH 1.0. Considering the intermediary, apical and basal regions of the filaments, the lamellar area was measured and the amount of mucous cells was counted. It was verified that oxygen, pH, and temperature decreased progressively as the water flew from one tank to another. Thus, an increase was realized of the amount of mucous cells and the lamellar area in T2, as well as a progressive decrease of these measures on the tanks which received water from T2. Moreover, detachment of the gill epithelium, cellular hyperplasia in the interlamellar space, and telangectasias were observed in the fishes from T2, T3 e T4. It was concluded that the aquatic environment in tanks sequentially interconnected with PVC pipes suffers alterations from one tank to another, as that physico-chemical fluctuations reflect on the behavior of the gill epithelium through variations of the lamellar area and the amount of mucous cells.

Abstract in English:

ABSTRACT.- Ribeiro M.G., Geraldo J.S., Langoni H., Lara G.H.B., Siqueira A.K., Salerno T. & Fernandes M.C. 2008. [Pathogenic microorganisms, somatic cell count and drug residues evaluation in organic bovine milk.] Microrganismos patogênicos, celularidade e resíduos de antimicrobianos no leite produzido no sistema orgânico. Pesquisa Veterinária Brasileira 29(1):52-58. Disciplina de Enfermidades Infecciosas dos Animais, Departamento de Higiene Veterinária e Saúde Pública, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista, Botucatu, SP 18618-000, Brazil. E-mail: mgribeiro@fmvz.unesp.br In last years increase the importance of milk quality and conditions of bovine milking. Simultaneously, increase the interest about organic milk and derivates. The aim of present study was investigate the milk pathogens, sensitivity and multiple drug resistance of isolates, somatic cell count and residues of drugs in milk, from cattle with and without mastitis, come from four little organic dairy farms in State of São Paulo, Brazil. Were used 148 cattle on the middle period of lactation. From these, two showed clinical mastitis, 72 subclinical mastitis and 74 without signs of mammary inflammation (control). Staphylococcus aureus (25.7%), Streptococcus spp. (21.4%), Corynebacterium bovis (12.9%), Streptococcus agalactiae (4.3%) and Staphylococcus spp (4,3%) were the more-frequent microorganisms isolated from animals with mastitis. Aspergillus spp. was isolated from one animal. Ceftiofur (95.2%), oxacillin (84.2%), gentamicin (76.3%) and cefoperazone (70.3%) were the more effective drugs. High resistance of isolates were found to penicillin (53.5%), ampicillin (41.6%) and neomycin (38.6%). Multiple drug resistance to three or more drug was observed in 40 (39.6%) isolates. Media of somatic cell count encountered in animals with mastitis and controls were 175,742.67cs/mL and 58,227.6 cs/mL, respectively. Antimicrobials residues in milk were detected in four (2.7%) animals. The present findings showed the low somatic cell count of animals, indicative of good quality of milk. However, pointed the need of control measures for contagious pathogens of bovine mastitis and more attention for prohibition of antimicrobial use in organic dairy farms.

• Organic milk mastitis bovine somatic cell count antimicrobials multiresistance

Abstract in Portuguese:

ABSTRACT.- Ribeiro M.G., Geraldo J.S., Langoni H., Lara G.H.B., Siqueira A.K., Salerno T. & Fernandes M.C. 2008. [Pathogenic microorganisms, somatic cell count and drug residues evaluation in organic bovine milk.] Microrganismos patogênicos, celularidade e resíduos de antimicrobianos no leite produzido no sistema orgânico. Pesquisa Veterinária Brasileira 29(1):52-58. Disciplina de Enfermidades Infecciosas dos Animais, Departamento de Higiene Veterinária e Saúde Pública, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista, Botucatu, SP 18618-000, Brazil. E-mail: mgribeiro@fmvz.unesp.br In last years increase the importance of milk quality and conditions of bovine milking. Simultaneously, increase the interest about organic milk and derivates. The aim of present study was investigate the milk pathogens, sensitivity and multiple drug resistance of isolates, somatic cell count and residues of drugs in milk, from cattle with and without mastitis, come from four little organic dairy farms in State of São Paulo, Brazil. Were used 148 cattle on the middle period of lactation. From these, two showed clinical mastitis, 72 subclinical mastitis and 74 without signs of mammary inflammation (control). Staphylococcus aureus (25.7%), Streptococcus spp. (21.4%), Corynebacterium bovis (12.9%), Streptococcus agalactiae (4.3%) and Staphylococcus spp (4,3%) were the more-frequent microorganisms isolated from animals with mastitis. Aspergillus spp. was isolated from one animal. Ceftiofur (95.2%), oxacillin (84.2%), gentamicin (76.3%) and cefoperazone (70.3%) were the more effective drugs. High resistance of isolates were found to penicillin (53.5%), ampicillin (41.6%) and neomycin (38.6%). Multiple drug resistance to three or more drug was observed in 40 (39.6%) isolates. Media of somatic cell count encountered in animals with mastitis and controls were 175,742.67cs/mL and 58,227.6 cs/mL, respectively. Antimicrobials residues in milk were detected in four (2.7%) animals. The present findings showed the low somatic cell count of animals, indicative of good quality of milk. However, pointed the need of control measures for contagious pathogens of bovine mastitis and more attention for prohibition of antimicrobial use in organic dairy farms.

Abstract in English:

ABSTRACT.- Corrêa A.M.R., Zlotowski P., Barcellos D.E.S.N., Cruz C.E.F. & Driemeier D. 2008. Gastric ulcers in pigs affected with postweaning multisystemic wasting syndrome. Pesquisa Veterinária Brasileira 28(12):601-605. Departamento de Patologia Clínica Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@ufrgs.br Samples of gastric lymph nodes and the stomachs from 24 pigs selected from herds affected by postweaning multisystemic wasting syndrome (PMWS) and sudden death associated with gastric ulcers were studied. Pigs were selected on the basis of unthriftiness, decreased feed intake, and wasting. The stomachs were opened, inverted, and classified into 0-3 score according the severity of the gross lesions present in pars oesophagica (non-glandulargastric mucosa). Selected samples were processed for paraffin embedding and stained with hematoxylin and eosin. Immunohistochemistry using anti-PCV2 (porcine circovírus type 2) antibody, anti-Helicobacter pylori antibody and a wide-spectrum anti-cytokeratin antibody was performed. Gross changes in pars oesophagea were classified according to the severity of lesions as score 3, 2, and 1 in 8, 6, 5 stomachs respectivelly. Microscopically, hyperplastic lymphoid follicles, lymphohistiocytic inflammatory infiltrates and focci of necrosis in the gastric mucosa were common findings. Large amounts of PCV2 antigen were observed in the cytoplasm and nuclei from intralesional cells and debris from the gastric glandular mucosal zone; however, in the fundus, anti-PCV2 immunostaining was restricted to the surface mucosal cells and foveolar compartment. All gastric lymph nodes were positive for PCV2 antigen. Anti-H. pylori immunostaining was seen in eleven cases, mainly in the antrum, on the mucosal surface and foveolar compartment. The association of the anti-PCV2 immunostaining with the glandular mucus-producing cells suggests a role for PCV2 as an additional factor for the swine ulcer development.

• Diseases of swine gastric ulcer PMWS PCV2 Helicobacter pylori

Abstract in Portuguese:

ABSTRACT.- Corrêa A.M.R., Zlotowski P., Barcellos D.E.S.N., Cruz C.E.F. & Driemeier D. 2008. Gastric ulcers in pigs affected with postweaning multisystemic wasting syndrome. Pesquisa Veterinária Brasileira 28(12):601-605. Departamento de Patologia Clínica Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@ufrgs.br Samples of gastric lymph nodes and the stomachs from 24 pigs selected from herds affected by postweaning multisystemic wasting syndrome (PMWS) and sudden death associated with gastric ulcers were studied. Pigs were selected on the basis of unthriftiness, decreased feed intake, and wasting. The stomachs were opened, inverted, and classified into 0-3 score according the severity of the gross lesions present in pars oesophagica (non-glandulargastric mucosa). Selected samples were processed for paraffin embedding and stained with hematoxylin and eosin. Immunohistochemistry using anti-PCV2 (porcine circovírus type 2) antibody, anti-Helicobacter pylori antibody and a wide-spectrum anti-cytokeratin antibody was performed. Gross changes in pars oesophagea were classified according to the severity of lesions as score 3, 2, and 1 in 8, 6, 5 stomachs respectivelly. Microscopically, hyperplastic lymphoid follicles, lymphohistiocytic inflammatory infiltrates and focci of necrosis in the gastric mucosa were common findings. Large amounts of PCV2 antigen were observed in the cytoplasm and nuclei from intralesional cells and debris from the gastric glandular mucosal zone; however, in the fundus, anti-PCV2 immunostaining was restricted to the surface mucosal cells and foveolar compartment. All gastric lymph nodes were positive for PCV2 antigen. Anti-H. pylori immunostaining was seen in eleven cases, mainly in the antrum, on the mucosal surface and foveolar compartment. The association of the anti-PCV2 immunostaining with the glandular mucus-producing cells suggests a role for PCV2 as an additional factor for the swine ulcer development.