Acellular pertussis vaccines in infants: evaluation of single component and two-component products.

Two acellular pertussis vaccines, one containing only LPF toxoid (25 micrograms) and the other containing LPF toxoid (25 micrograms) and FHA (25 micrograms) and each combined with diphtheria and tetanus toxoids, were evaluated in two groups of 25 infants. A third group of 25 infants served as controls and received a DTP whole-cell pertussis vaccine. Infants given either acellular pertussis vaccine had significantly fewer local and systemic reactions than infants given whole-cell vaccine. Among the three vaccine groups, infants given the LPF vaccine (single component) had the highest concentration of antibody to LPF after three immunizations. Infants receiving the LPF/FHA vaccine (two-component) had the highest concentration of antibody to FHA after three immunizations. Infants vaccinated with the two-component vaccine had a significantly lower serological response to LPF than infants given the single component vaccine, as measured by either enzyme-linked immunosorbent assay or CHO cell assay. Further studies are necessary to determine why differences in immunogenicity of the two investigational vaccines occurred.

Enzyme-linked immunosorbent assay with major outer membrane proteins of Brucella melitensis to measure immune response to Brucella species.

We developed an enzyme-linked immunosorbent assay (ELISA) system to measure human immunoglobulin G (IgG) and IgM response to the major outer membrane proteins of Brucella melitensis. The ELISA was more sensitive in detecting antibody than a standard microagglutination (MA) test with B. abortus antigen. Of 101 sera from persons with suspected brucellosis, 79 (78.2%) gave ELISA IgM titers greater than or equal to the B. abortus MA titer without 2-mercaptoethanol (2ME), which measures both IgM and IgG. Of the 101 sera, 97% gave ELISA IgG titers greater than or equal to the MA with 2ME titer. A total of 58 sera, drawn from 11 human patients from 1 to 29 weeks after onset of brucellosis, gave higher geometric mean titers for the ELISA IgG test than for the MA with 2ME test. These 58 sera also gave ELISA IgM geometric mean titers that were greater than or within one doubling dilution of the geometric mean titers of MA without 2ME. In addition to detecting antibody response to B. abortus, B. melitensis, and B. suis, the ELISA was sensitive to antibody response to human and canine infections with B. canis. The B. canis antibody response is not detected by the MA test with B. abortus antigen. The ELISA, with a standard preparation of major outer membrane proteins of B. melitensis as antigen, appears to be useful in measuring antibody response in humans to infections by all species of Brucella known to infect humans.

New blood culture medium.

A growth medium with a specific oxidation-reduction potential containing peptone, dextrose, sodium succinate, sodium lactate, gelatin, sodium bicarbonate and blue tetrazolium, an indicator dye, in a tris(hydroxymethyl)aminomethane buffer was used to detect the presence of microorganisms in blood. The procedure involved the introduction of blood (and bacteria) into the growth medium with the dye in its colorless state. As the bacteria grew, they converted the dye to a visible blue color (formazan) with their reductases. The growth medium served as its own contamination control, since microbial growth and be detected by a color change before it was used for blood culture. The experiments described herein demonstrate that the composition of this medium (with the dye) provides a unique system that is able to make a reliable and rapid detection of both gram-positive and gram-negative microorganisms and yeasts (Candida albicans) commonly associated with bacteremia.

Effect of age, sex, and intestinal flora on the induction of colon tumors in rats.

Germfree and conventional Sprague-Dawley rats were assessed for their susceptibility to intrarectally injected N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) or N-methyl-N-nitrosourea (MNU). Adenocarcinoma of the colon was induced in germfree and conventional rats by both MNNG and MNU. The colons of germfree rats were more susceptible to the direct-acting carcinogens, as manifested by earlier morbidity and development of colon tumors (50% tumors within 30-35 wk), than were those of conventional rats (50% colon tumors within 48-50 wk). Germfree and conventional male rats were more susceptible to the carconogens than were their female germfree and conventional counterparts. Young (30 days old at the start of the experiment) germfree rats developed colon tumors more quickly (15-20 wk) than did older (60 days) germfree rats after intrarectal injections of MNNG. No colon tumors were observed in germfree and conventional rats after 75 weekly intrarectal injections with a buffer. Transplantation of an adenocarcinoma induced with MNU in a female rat to germfree and conventional rats showed that it was easily transplantable, required no immunosuppression, and had essentially the same morphologic characteristics as did the primary tumor.

Aflatoxin can be degraded by the mycelium of aspergillus parasiticus.

Aflatoxins B1, B2, G1, and G2 were degraded by the 8- and 16-day old but not by the 4-day old mycelium of a toxigenic strain of Aspergillus parasiticus. The 16-day old mycelium degraded the toxin more rapidly than did the 8-day old mycelium. Degradation of toxin by the mycelium was similar at pH 2.5 and 6.0.

Production of aflatoxin and its partition between the medium and the mycelium of Aspergillus parasiticus during incubation under various conditions.

Spores of an aflatoxigenic strain of Aspergillus parasiticus were inoculated into a glucose-salts medium which was incubated with and without shaking at 28 degrees C for 15 days. Without shaking, maximal production of total aflatoxin and aflatoxins B1, G1, and G2 occurred at 5 days, whereas the maximal amount of B2 appeared after 7 days. Initially approximately 5% of the total toxins appeared in the mycelium but this increased to more than 60% after 5 days. Shaking of cultures during incubation served to reduce production of total aflatoxin and of each of the individual toxins. The maximal amount of total aflatoxin and of toxins B1 and G1 appeared in shaken cultures after 5 days, whereas 8 and 11 days were needed to obtain maximal amounts of B2 and G2, respectively. The mycelium of shaken cultures initially retained approximately 50% of the total aflatoxin and this increased to about 80% as the incubation progressed. Very little aflatoxin was synthesized at 35 and 45 degrees C and production of total aflatoxin and of each individual toxin was less at 15 degrees C than at 25 or 28 degrees C. When the medium contained 0.5 to 50% glucose, maximal amounts of total aflatoxin and of aflatoxins B1, G1 and G2 appeared in the presence of 30% glucose; only 20% glucose was needed to obtain the greatest amount of B2. The mycelium retained approximately 50% of total aflatoxin when the medium contained 5 to 20%. Neither aflatoxin G1 nor G2 were detected when the medium contained 0.05% ammonium sulfate and only B1, B2, and G1 appeared in the medium with 0.1% of the salt. Maximal production of each individual aflatoxin and of total aflatoxin occured with 1% of ammonium sulfate in the medium. The proportion of total aflatoxin retained by the mycelium decreased from 83 to 37% as the amount of ammonium sulfate in the medium was increased from 0,05 to 10%.