ABSTRACT
The aim of the study presented here was to analyze parapertussis infections occurring in the region of Warsaw, Poland, between 1995 and 2002 and to characterize the Bordetella parapertussis isolates using macrorestriction digestion of genomic DNA. Fourteen of the 17 patients with confirmed Bordetella parapertussis infection were between the ages of 5 and 9 years, and 14 had documented vaccination with the diphtheria-tetanus-pertussis whole-cell vaccine; for the remaining three patients vaccination data was not available. DNA fingerprinting performed using recommended standard methodology revealed a close relationship between the examined strains and reference strain ATCC 9305, thus confirming the conservative nature of Bordetella parapertussis.
Subject(s)
Bordetella Infections/diagnosis , Bordetella Infections/epidemiology , Bordetella parapertussis/isolation & purification , Age Distribution , Anti-Bacterial Agents/therapeutic use , Bordetella Infections/drug therapy , Child , Child, Preschool , DNA, Bacterial/analysis , Female , Humans , Incidence , Male , Poland/epidemiology , Polymorphism, Restriction Fragment Length , Retrospective Studies , Risk Factors , Sex Distribution , Urban PopulationABSTRACT
The aim of this study was to compare bacteria growth and pertussis antigens (pertussis toxin-PT, filamentous haemagglutinin-FHA and endotoxin-LPS) production by 11 Bordetella pertussis strains. A synthetic Stainer-Scholte culture medium supplemented with (2,6-0-dimethyl) beta-cyclodextrin (heptakis) or methylcellulose (for greater PT and FHA production) and solid modified Cohen-Wheeler medium (for LPS isolation) were used. Our results demonstrated that heptakis and methylcellulose were more effective for antigens production than for bacterial growth. It was interesting that these stimulated substances supported the bacterial growth from small inocula. The investigated strains differed in PT, FHA and LPS production. The best PT producer was the B. pertussis 134 strain, the worst B. pertussis 2897. The differences in FHA production are not as big as in PT production, but the B. pertussis 2897 was also the worst FHA producer. Isolated LPS consists of dry bacteria pellet ranging from 1,9% (B.p. Tohama) to 5,6% (B.p. 3803 strain). No great differences in serological activity of LPS isolated from different strains were observed. In the haemagglutination inhibition test the endotoxin isolated from B.p. 509 and B.p. Tohama strains showed the highest activity.
Subject(s)
Bacterial Toxins/biosynthesis , Bordetella pertussis/growth & development , Bordetella pertussis/metabolism , Adhesins, Bacterial/biosynthesis , Bordetella pertussis/pathogenicity , Cyclodextrins , Endotoxins/biosynthesis , Hemagglutination Tests , Hemagglutinins/biosynthesis , Lipopolysaccharides/biosynthesis , Methylcellulose , Pertussis Toxin , Species Specificity , Virulence , Virulence Factors, Bordetella/biosynthesisABSTRACT
Acellular preparations were studied which were obtained by a method of acidic extraction from culture of strain Tohama of Bordetella pertussis. Obtained preparations EM I and EM II were exhibiting property of leukocytosis stimulation and were respectively of following values: 43.5 and 19 u LPF/mg of protein. They also caused hemagglutination (150 and 19 u HA/mg of protein). Both preparations were exhibiting protective effect in intracerebral test performed on mice. After differentiation of EM I preparation by preparative centrifugation in saccharose gradient two fractions were obtained, of which lighter was exhibiting high hemagglutinating activity and leukocytosis stimulation (198 u LPF/mg of protein). Heaver fraction contained mainly endotoxin. Applied method permitted for partial purification of acellular preparation of B. pertussis; mainly decrease in its content of endotoxin responsible for high reactogenicity of pertussis vaccines.
