RESUMO
We developed and optimized a simple, efficient and inexpensive method for in vitro culture of peripheral blood lymphocytes from the Brazilian tortoise Chelonoidis carbonaria (Testudinidae), testing various parameters, including culture medium, mitogen concentration, mitotic index, culture volume, incubation time, and mitotic arrest. Peripheral blood samples were obtained from the costal vein of four couples. The conditions that gave a good mitotic index were lymphocytes cultured at 37°C in minimum essential medium (7.5 mL), with phytohemagglutinin as a mitogen (0.375 mL), plus streptomycin/penicillin (0.1 mL), and an incubation period of 72 h. Mitotic arrest was induced by 2-h exposure to colchicine (0.1 mL), 70 h after establishing the culture. After mitotic arrest, the cells were hypotonized with 0.075 M KCl for 2 h and fixed with methanol/acetic acid (3:1). The non-banded mitotic chromosomes were visualized by Giemsa staining. The diploid chromosome number of C. carbonaria was found to be 52 in females and males, and sex chromosomes were not observed. We were able to culture peripheral blood lymphocytes of a Brazilian tortoise in vitro, for the preparation of mitotic chromosomes.
Assuntos
Técnicas de Cultura de Células , Cromossomos/genética , Meios de Cultura/química , Linfócitos/citologia , Tartarugas/genética , Animais , Corantes Azur/análise , Brasil , Bandeamento Cromossômico , Cromossomos/química , Colchicina/farmacologia , Feminino , Fixadores , Humanos , Linfócitos/efeitos dos fármacos , Masculino , Mitógenos/farmacologia , Índice Mitótico , Fito-Hemaglutininas/farmacologiaRESUMO
Azo, anthroquinone and triphenylmethane dyes are the major classes of synthetic colourants, which are difficult to degrade and have received considerable attention. Congo red, a diazo dye, is considered as a xenobiotic compound, and is recalcitrant to biodegradative processes. Nevertheless, during the last few years it has been demonstrated that several fungi, under certain environmental conditions, are able to transfer azo dyes to non toxic products using laccases. The aim of this work was to study the factors influencing mycoremediation of Congo red. Several basidiomycetes and deuteromycetes species were tested for the decolourisation of Congo red (0.05 g/l) in a semi synthetic broth at static and shaking conditions. Poor decolourisation was observed when the dye acted as the sole source of nitrogen, whereas semi synthetic broth supplemented with fertilizer resulted in better decolourisation. Decolourisation of Congo red was checked in the presence of salts of heavy metals such as mercuric chloride, lead acetate and zinc sulphate. Decolourisation parameters such as temperature, pH, and rpm were optimized and the decolourisation obtained at optimized conditions varied between 29.25- 97.28 percent at static condition and 82.1- 100 percent at shaking condition. Sodium dodecyl sulphate polyacrylamide gel electrophoretic analysis revealed bands with molecular weights ranging between 66.5 to 71 kDa, a characteristic of the fungal laccases. High efficiency decolourisation of Congo red makes these fungal forms a promising choice in biological treatment of waste water containing Congo red.
Assuntos
Basidiomycota , Corantes Azur/análise , Lacase/análise , Vermelho Congo/análise , Xenobióticos/análise , Biodegradação Ambiental , Microbiologia Ambiental , Métodos , MétodosRESUMO
The aim of this study was to detect the presence of Helicobacter infection in Thoroughbred in Venezuela. One hundred and thirty six Thoroughbreds with signs for Equine gastric ulcer syndrome (EGUS) were studied in the National Race Track La Rinconada Caracas Venezuela. Necropsies were performed in all horses and samples of gastric tissue were collected. The tissue samples were fixed in formalin and processed by conventional H&E techniques. Additionally, the special staining procedure of Warthin-Starry, Giemsa and Blue Toluidine were also carried out. Ureasa test: rapid ureasa test (commercial kit) was performing of gastric tissue. The damage of the gastric mucosae in the animals was categorized as 2/136 Grade 0 Epithelium is intact throughout; no hyperemia, no hyperkeratosis. Grade 1 Mucosa is intact but there are areas of hyperemia and/or hyperkeratosis 26/136. Grade 2 small, single or multi-focal erosions or ulcers 38/136. Grade 3 Large, single or multi-focal ulcers, or extensive erosions and sloughing 36/136.Grade 4 Extensive ulcers, with areas with deep submucosal damage 34/136. Using the Warthin-Starry special stain, Giemsa and Blue Toluidine with spiral-shaped bacteria were evidenced found in 52, Grade 1: 13/52, Grade 2: 10, Grade 3: 16, Grade 4: 13. There seemed to be no association between Warthin-Starry, Giemsa and Blue Toluidine stain with the grade and location of lesions. Urease activity was then demonstrated in fifty-two equine. To conclude, we detected the presence of Helicobacter Like Organisms in the gastric mucosa from thoroughbred horses with EGUS (AU)
Assuntos
Animais , Helicobacter , Helicobacter/isolamento & purificação , Cloreto de Tolônio , Corantes Azur/análise , Corantes Azur , Cavalos , Coleta de Tecidos e Órgãos/métodos , Coleta de Tecidos e Órgãos/veterináriaRESUMO
The aim of this study was to detect the presence of Helicobacter infection in Thoroughbred in Venezuela. One hundred and thirty six Thoroughbreds with signs for Equine gastric ulcer syndrome (EGUS) were studied in the National Race Track La Rinconada Caracas Venezuela. Necropsies were performed in all horses and samples of gastric tissue were collected. The tissue samples were fixed in formalin and processed by conventional H&E techniques. Additionally, the special staining procedure of Warthin-Starry, Giemsa and Blue Toluidine were also carried out. Ureasa test: rapid ureasa test (commercial kit) was performing of gastric tissue. The damage of the gastric mucosae in the animals was categorized as 2/136 Grade 0 Epithelium is intact throughout; no hyperemia, no hyperkeratosis. Grade 1 Mucosa is intact but there are areas of hyperemia and/or hyperkeratosis 26/136. Grade 2 small, single or multi-focal erosions or ulcers 38/136. Grade 3 Large, single or multi-focal ulcers, or extensive erosions and sloughing 36/136.Grade 4 Extensive ulcers, with areas with deep submucosal damage 34/136. Using the Warthin-Starry special stain, Giemsa and Blue Toluidine with spiral-shaped bacteria were evidenced found in 52, Grade 1: 13/52, Grade 2: 10, Grade 3: 16, Grade 4: 13. There seemed to be no association between Warthin-Starry, Giemsa and Blue Toluidine stain with the grade and location of lesions. Urease activity was then demonstrated in fifty-two equine. To conclude, we detected the presence of Helicobacter Like Organisms in the gastric mucosa from thoroughbred horses with EGUS