Avaliação clínica da infecção experimental de bezerros com Salmonella Typhimurium / Clinical evaluation of experimental Salmonella Typhimurium-infection in calves

The clinical conditions of healthy calves infected with 10(9) CFU of Salmonella Typhimurium were evaluated and the viability of the experimental model in disease induction was verified. Twelve male Holstein calves, 10 to 15-days old, were examined. They were allocated into two groups, control and experimentally infected. Animals were submitted to clinical examination after inoculation and at every 24 hours, during six days after infection. Samples of rectal swabs were collected for Salmonella Typhimurium isolation. Results showed that all animals had different degrees of diarrhea, with mucus and bleeding, 24 to 72 hours after experimental infection with Salmonella Typhimurium, accompanied by fever, but no signs of sepsis. The isolation of Salmonella Typhimurium from rectal swabs occurred 24 hours after the infection and there were no deaths during the experiment, which means that this sorovar has an epidemiological importance related to the intermitent or continuous elimination of the bacterium. Results also showed that the oral administration of 10(9) CFU of Salmonella Typhimurium induces clinical signs of salmonellosis in 10 to 15-day-old calves.

Large unilamellar vesicles as trehalose-stabilised vehicles for vaccines: storage time and in vivo studies.

Liposomes, as a pharmaceutical formulation must display a long shelf life. The recombinant heat-shock protein from Mycobacterium leprae (18-kDa hsp) or its N-acylated derivative, when entrapped within or externally associated with large unilamellar vesicles, acts as a T-epitope source. Freeze-fracture electron microscopy shows unequivocally that trehalose avoids aggregation and fusion of these vesicles. Formulations containing trehalose retained up to 98% of the entrapped protein. The highest antibody level is obtained with formulations containing trehalose. The adjuvant effect depends on the liposomal membrane integrity.

Large unilamellar vesicles as trehalose-stabilised vehicles forvaccines storage time and in vivo studies

Liposomes, as a pharmaceutical formulation must display a long shelf life. The recombinant heat-shock protein fromMycobacterium leprae (18-kDa hsp) or its N-acylated derivative, when entrapped within or externally associated with largeunilamellar vesicles, acts as a T-epitope source. Freeze-fracture electron microscopy shows unequivocally that trehaloseavoids aggregation and fusion of these vesicles. Formulations containing trehalose retained up to 98% of the entrappedprotein. The highest antibody level is obtained with formulations containing trehalose. The adjuvant effect depends on theliposomal membrane integrity.

Conformational stability and antibody response to the 18kDa heat-shock protein formulated into different vehicles.

Protein stability is one of the most important obstacles for successful formulation in the development of new-generation vaccines. Here, the 18kDa heat-shock protein (18kDa-hsp) was chemically modified though conjugation with bovine serum albumin or by esterification with N-hydroxysuccinimide ester of palmitic acid. The biologically active conformation of the protein was preserved after chemical modification. The immune responses to the recombinant 18kDa-hsp from Mycobacterium leprae were studied in different presentations: free, copolymerized with bovine serum albumin in aggregates (18kDa-hsp-BSA), and either surface linked to liposomes or entrapped into liposomes. Measuring the antibody production of immunized genetically selected mice has compared the adjuvant effects of liposomes and proteic copolymer. Among the two liposome preparations, the strongest response was obtained with the surface-exposed antigen-liposomes. The copolymer 18kDa-hsp-BSA conferred a high titer of antibody in injected mice, and persisted 70 d after immunization. This approach should prove very useful for designing more effective vaccines by using 18kDa-hsp as carrier protein.

Microquantification of proteins with low chromophore content

The advantages of the intrinsic fluorescence of the tryptophan and the absorbance of the methionine residues of the 18 kDa-hsp - a recombinant protein from Mycobacterium leprae - was exploited here to develop a sensitive and low costs method for protein assaying. They presented linearity between 3 and 1000 ìg of protein. The correlations between intrinsic fluorescence or absorbance at 230 nm and protein contents were both superiors to 0.99. These methods can be extended to others proteins with low aromatic residue contents

Estenosis Hipertrófica Congénita del Píloro / Congenital Hipertrophics Stenosis of Pylorus

Se analizan los aspectos clínicos y terapéuticos de 44 casos de estenosis hipertrófica congénita del píloro.El estudio demostró predominio del sexo masculino y su máxima presentación entre la segunda y tercera semana de edad postnatal. Los síntomas y signos más frecuentes fueron: vómitos en proyectil, palpación de la oliva pilórica y ondas peristálticas visibles. Todos los pacientes fueron tratados quirúrgicamente, siendo las complicaciones más frecuentes perforación duodenal y pilórica. No hubo mortalidad.