Phenotypical and molecular characterization of Acinetobacter spp. isolated from a pharmaceutical facility.

Characterizing microorganisms according to different criteria is useful when investigating sources of microbiological contamination in the pharmaceutical industry. The aim of this study was to characterize 38 Acinetobacter baumannii complex strains isolated from a biopharmaceutical industry by 16S rRNA sequencing, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS), multilocus sequence typing (MLST), antimicrobial susceptibility profile, biofilm formation, and sensibility to disinfectants. Thirty-three (86.9%) strains were identified by 16S rRNA gene sequencing as A. seifertii/pitti/nosocomialis/lactucae, four (10.5%) as A. baumannii, and one (2.6%) as A. vivianii/courvalini. MALDI-TOF/MS did not identify one strain, and incorrectly identified 30/37 (81.1%) strains as A. baumannii. Strains were assigned to 12 different STs, of which nine were newly defined in this study (STs 2091-2099). Twenty-six (68.4%) strains showed resistance to amikacin and gentamicin. Thirty-three (86.8%) strains were classified as moderately or strongly adherent on polystyrene. Alcohol 70%/15 min and quaternary ammonium 0.08%/20 min were not able to eliminate the biofilm formed, but sodium hypochlorite 0.1%/15 min was efficient. In conclusion, improved methods are needed to improve the identification of Acinetobacter strains in pharmaceutical industries. This organism is of particular concern as it forms recalcitrant biofilms, leading to persistence in the manufacturing environment and increased risk of product contamination.

Establishment of certified reference material for the potency assay in yellow fever vaccine quality control, in accordance with International Standards Organization guidance.

The attenuated yellow fever vaccine (YFV) is offered free of charge to the Brazilian population through the National Immunization Program (NIP). One of the specifications for quality control analyses of the vaccine is the potency determination. This test determines the number of plaque forming units (PFU) in Vero cells. In order to validate the results, the reference material (RM) is analysed in parallel with an established reference vaccine. The aim of this study was to establish certified RM for use as an internal control in the potency assay for the production chain of YFV. The candidate RM homogeneity and stability were determined, and characterized by a collaborative study for further certification. The RM was considered sufficiently homogeneous with average 4.68 log10 IU/HD and stable at (-20 ± 10) ºC and (22.5 ± 2.5) ºC for 715 and 183 days, respectively. When reconstituted and stored in aliquots of 0.6 mL, it was stable at (-20 ± 10) ºC for eight days. But it was not stable at (5 ± 3) ºC for three days. In a collaborative study, two independents' laboratories gave an averaged value of 4.56 ± 0.030 log10 IU/HD. After determining the expanded uncertainty of homogeneity, stability, and characterization, the certified RM lot: 195VFA020Z presented a property value of 4.56 ± 0.22 log10 IU/HD. It was concluded that the new certified RM can be used in routine analysis of a YFV producer, since it has its property value established and it is stable. The possibility of using it in aliquots after reconstitution will also allow the RM to have a much longer shelf life.

Efeitos da vacina pertussis sobre monooxigenases hepáticas de camundongos / Hepatic pertussis effect of the vaccine on monooxigenases of mice

Neste trabalho investigamos os efeitos dos da vacina pertussis sobre a atividade de monooxigenases hepáticas em camundongos C57BL6, machos e fêmeas, ao longo de 3 semanas que se seguiram ao tratamento e comparamos também os efeitos da vacina pertussis com os da endotoxina da bactéria E.coli na modulação das CYPs hepáticas. Após o tratamento por via intraperitoneal com a vacina pertussis (1, 2, 5, 7, 14 e 21 dias após a administração) ou com o LPS de E.coli (1 e 7 dias após a administração), os animais foram sacrificados por deslocamento cervical sendo o fígado imediatamente retirado e a fração microssomal preparada por homogeneização e duas ultracentrifugações (100.000 g por 60 minutos) sucessivas. As atividades de reações marcadoras de isoenzimas CYPs - das subfamílias 1A (EROD), 2B (BROD) e 2E (PNP hidroxilase) – foram medidas nos microssomos hepáticos de camundongos machos e fêmeas tratados com a vacina, com o LPS de E.coli, ou com seus respectivos controles. Os resultados mostraram que a vacina pertussis causou acentuada depressão das atividades de monooxigenases hepáticas (EROD, BROD e PNP hidroxilase) 24 horas após o tratamento. Nos dias subseqüentes houve uma recuperação parcial que foi no entanto seguida de nova queda da atividade que atingiu os níveis mais baixos por volta do 7° dia após a injeção, permanecendo deprimida em relação aos controles pelo menos até o 21° dia pós-administração. As fêmeas foram mais susceptíveis aos efeitos inibitórios da vacina sobre as atividades das monooxigenases. A diferença entre o curso temporal da inibição observada nos animais tratados com a vacina pertussis e com o LPS de E.coli, sugere que a modulação da atividade de CYPs pela vacina pertussis, na sua fase inicial, seria mediada pela endotoxina nela contida (resposta rápida, 24 – 48 horas), resultando porem a depressão posterior (resposta máxima aos 7 dias) de efeitos da toxina pertussis.