Resultado da pesquisa (3)

Termo utilizado na pesquisa cultivation

#1 - Isolation, cultivation and immunofluorescence characterization of lamellar keratinocytes from equine hoof by using explants

Abstract in English:

The importance of the hoof to the horse health is clear, and the current knowledge regarding the cellular aspects of hoof keratinocytes is poor. Studies on equine keratinocyte culture are scarce. Developing keratinocyte cultures in vitro is a condition for studies on molecular biology, cell growth and differentiation. Some methods have already been established, such as those for skin keratinocyte culture. However, few methodologies are found for lamellar keratinocytes. The objective of this study was to standardize the equine hoof keratinocyte isolation and cultivation, and then characterize the cell immunophenotype. For this, the primary culture method used was through explants obtained from three regions of the equine hoof (medial dorsal, dorsal, and lateral dorsal). After the cell isolation and cultivation, the cell culture and its explants were stained with anti-pan cytokeratin (pan-CK) (AE1/AE3), vimentin (V9), p63 (4A4), and Ki-67 (MIB-1) antibodies. Cells were grown to third passage, were positive for pan-CK, p63 and Ki-67, and few cells had vimentin positive expression. As for the explants, the epidermal laminae were not stained for vimentin or Ki-67. However, some cells presented positive pan-CK and p63 expression. This study demonstrated the viability of lamellar explants of equine hooves as a form of isolating keratinocytes in primary cultures, as well as characterized the proliferation ability of such keratinocytes in monolayers.

Abstract in Portuguese:

É notória a importância do casco na saúde dos equinos, mas o conhecimento em nível celular é pouco entendido. Estudos envolvendo o cultivo de queratinócitos equinos são escassos. Sabe-se que o desenvolvimento de cultivos de queratinócitos in vitro é uma condição para estudos sobre a biologia molecular, crescimento e diferenciação celular. Alguns métodos já estão estabelecidos, como para cultivo de queratinócitos de pele, mas poucas metodologias são encontradas para queratinócitos lamelares. O objetivo desse estudo foi padronizar o cultivo de queratinócitos provenientes de casco equino visando futuramente associar ao estudo da medicina regenerativa para assim estabelecer um modelo experimental in vitro e indicar o uso criterioso de terapias regenerativas para a laminite equina. Desta forma, o cultivo em monocamada e a caracterização de queratinócitos lamelares foram realizados. Para isso, o método de cultura primária utilizado foi através de explantes obtidos de três regiões do casco (dorso-medial, dorsal e dorso-lateral). As células foram caracterizadas para os marcadores anti pan-cytokeratin (AE1/AE3), vimentin (V9), p63 (4A4) e Ki-67 (MIB-1) nos cultivos e nos explantes. As  células foram cultivadas até terceira passagem, tendo marcação positiva para pan-CK, p63 e Ki-67 e fraca marcação para vimentina. Já as lâminas epidermais não tiveram marcação de vimentin e Ki-67, porém marcaram acentuadamente para pan-CK e p63. Este estudo demonstrou a exiquibilidade do uso de explantes lamelares do casco de equinos, como forma de isolamento de queratinócitos em cultivos primários, bem como caracterizou a habilidade de proliferação desses queratinócitos em monocamada.


#2 - Cinética do crescimento de Borrelia burgdorferi em diferentes meios de cultivo

Abstract in English:

Oliveira A., Fonseca A.H., Ishikawa M.M. & Yoshinari N.H. 2004. [Cinetic growth of Borrelia burgdorferi (Spirochaetaceae) in different culture media.] Cinética do crescimento de Borrelia burgdorferi em diferentes meios de cultivo. Pesquisa Veterinária Brasileira 24(2):61-64. Depto Epidemiologia e Saúde Pública, Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, Seropédica, RJ 23890-000, Brazil. E-mail: adivaldo@ufrrj.br The cinetic of growth of Borrelia burgdorferi was studied during a 3-month period, using the following 8 culture media: (1) rabbit serum BSK, (2) swine serum BSK, (3) swine serum BSK+5 fluorouracil, (4) PMR, (5) CTB, (6) Dubos, (7) Brucella broth and (8) BHI. All media were prepared aseptically and were maintained in culture tubes of 10 ml capacity. For each medium, the inoculum was standardized to contain initially 102 spirochetes for each 0.1 ml of culture. The growth was monitorized by counting the total number of spirochetes in 0.1ml of medium in a dark field microscope, using a 10x30 mm cover slip. For the first 12 days, counting was done each 24 hours, and afterwards once a week during 14 weeks. There occurred growth of B. burgdorferi in all tested media, with the best performance of three of them: BSK with rabbit serum, BSK swine serum + 5 fluorouracil, and CTB medium. Growth of B. burgdorferi was seen from the 4th week on, reaching its maximum within 8-12 weeks, depleting the culture medium after this time. Cystic forms of B. burgdorferi were observed with all tested media.

Abstract in Portuguese:

Oliveira A., Fonseca A.H., Ishikawa M.M. & Yoshinari N.H. 2004. [Cinetic growth of Borrelia burgdorferi (Spirochaetaceae) in different culture media.] Cinética do crescimento de Borrelia burgdorferi em diferentes meios de cultivo. Pesquisa Veterinária Brasileira 24(2):61-64. Depto Epidemiologia e Saúde Pública, Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, Seropédica, RJ 23890-000, Brazil. E-mail: adivaldo@ufrrj.br The cinetic of growth of Borrelia burgdorferi was studied during a 3-month period, using the following 8 culture media: (1) rabbit serum BSK, (2) swine serum BSK, (3) swine serum BSK+5 fluorouracil, (4) PMR, (5) CTB, (6) Dubos, (7) Brucella broth and (8) BHI. All media were prepared aseptically and were maintained in culture tubes of 10 ml capacity. For each medium, the inoculum was standardized to contain initially 102 spirochetes for each 0.1 ml of culture. The growth was monitorized by counting the total number of spirochetes in 0.1ml of medium in a dark field microscope, using a 10x30 mm cover slip. For the first 12 days, counting was done each 24 hours, and afterwards once a week during 14 weeks. There occurred growth of B. burgdorferi in all tested media, with the best performance of three of them: BSK with rabbit serum, BSK swine serum + 5 fluorouracil, and CTB medium. Growth of B. burgdorferi was seen from the 4th week on, reaching its maximum within 8-12 weeks, depleting the culture medium after this time. Cystic forms of B. burgdorferi were observed with all tested media.


#3 - Outbreaks of periodontitis in calves ("Cara inchada") related to soil cultivation

Abstract in English:

"Cara inchada" of cattle (CI) is a periodontitis which occurs in young animals during tooth eruption and is related to tree clearance atid sowing pasture grasses in certain areas of Central Brazil once covered by forest or savanna vegetation. The findings of outbreaks of CI on two farms in the State of São Paulo are reported. In these outbreaks the calves showed severe emaciation, diarrhea, various periodontal lesions, halitosis and loss of premolar teeth of the upper jaw. On one farm the disease occurred after feeding sugar cane harvested from a newly cultivated area, in which the prevalence of periodontal lesions was 61.5% and mortality 22.1%. On the other farm, the outbreak occurred after cattle grazed on recently improved pastures. In the dry season the animals were also fed supplements of forage and silage grown on the sarne farm. The prevalence of CI in the latter was 51.9% and mortality 3.6%. It is concluded that there is a relationship between new outbreaks of periodontitis and soil and pasture improvement, i.e. ploughing, harrowing, application of lime and fertilizers.

Abstract in Portuguese:

"Cara inchada" dos bovinos (CI) é uma periodontite que os acomete na fase de dentição e está associada à formação de pastagens em determinadas áreas antes cobertas de mata ou com vegetação de cerrado no Brasil Central. Foram registrados dois surtos da doença em propriedades situadas no Estado de São Paulo, após a reforma de áreas anteriormente positivas para CI, e utilização das forrageiras cultivadas na alimentação animal. Nas duas propriedades surtos da doença haviam sido observados há mais de 10 anos atrás, após a formação inicial das pastagens em área de cerrado; no entanto, nos últimos anos não havia mais registros da ocorrência da periodontite. No ressurgimento da doença os bezerros apresentaram emagrecimento acentuado, diarréia, lesões peridentárias de diferentes intensidades, odor fétido bucal e perda dos dentes premolares decíduos maxilares. Numa das propriedades a doença ocorreu após a utilização de cana-de-açúcar, cultivada no ano anterior, na alimentação dos bezerros. A prevalência de animais com lesões peridentárias foi de 61,5%, com coeficiente de mortalidade de 22,1%. Na outra propriedade o surto ocorreu após a reforma dos pastos e das áreas destinadas à capineira e silagem de milho. A prevalência da doença foi de 51,9%, com mortalidade de 3,6%. Numa análise retrospectiva pode-se associar o ressurgimento da periodontite em bezerros nas duas propriedades com a reforma das áreas através de aragem, gradeação, calagem e adubação do solo.


Colégio Brasileiro de Patologia Animal SciELO Brasil CAPES CNPQ UNB UFRRJ CFMV