A rapid microprocedure for isolating detergent (sodium N-lauroyl sarcosinate)-insoluble major outer membrane proteins from Haemophilus species produced results qualitatively identical to those obtained with a commonly used preparative isolation procedure. Proteins isolated by both procedures were compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis after staining with Coomassie brilliant blue R-250. The time for outer membrane protein isolation was substantially reduced with the rapid procedure, allowing a larger number of membrane preparations to be obtained rapidly for routine analysis.

Brazilian purpuric fever (BPF) is a recently recognized fulminant pediatric disease characterized by fever, with rapid progression to purpura, hypotensive shock, and death. BPF is usually preceded by purulent conjunctivitis that has resolved before the onset of fever. Both the conjunctivitis and RPF are caused by Haemophilus influenzae biogroup aegyptius (formerly called H. aegyptius). Isolates from 15 BPF cases, mainly from blood or hemorrhagic cerebrospinal fluid, case-associated isolates from 42 persons in towns where BPF cases occurred, and control strains from 32 persons in towns without BPF cases were characterized biochemically, genetically, and epidemiologically. Results indicated that a single clone was responsible for all BPF cases identified in six Brazilian towns from 1984 through 1986. All of 15 (100%) case strains were the same clone as was 1 of 32 (3%) control strains (P = <10-8). Isolates of the clone were preferentially intrarelated by DNA hybridization (99% relatedness, hydroxyapatite method at 60 and 75°C) and were separable from other H. influenzae biogroup aegyptius strains (approximately 90% relatedness at 60°C and 82% relatedness at 75°C). All isolates of the BPF clone and no other strains contained a 24-megadalton plasmid of restriction endonuclease type 3031, were of a single multilocus enzyme mobility type, were of a single sodium dodecyl sulfate-polyacrylamide gel electrophoresis type, and were in one of two ribosomal DNA restriction patterns. Ail BPF clone isolates reacted with monoclonal antibodies produced from a case strain; only 3 of 62 (5%) other strains reacted with this monoclonal antibody. Ninety percent of BPF clone strains and 27% of other strains were relatively resistant to sulfamethoxazole-trimethoprim.

Seventeen strains of Haemophilus ducreyi were isolated from genital lesions which were negative for syphilis by dark-field examination. Media used for primary isolation at various times during the study were enriched chocolate agar, chocolate agar plus vancomycin (3 microgram/ml), rabbit blood agar plus vancomycin (3 micrograms/ml), fetal bovine serum agar, and fetal bovine serum agar plus vancomycin (3 micrograms/ml). H. ducreyi was isolated on chocolate agar plus vancomycin from 10 of 14 patients found to be positive on one or more media, on rabbit blood agar plus vancomycin from 16 of 17 patients, and on fetal bovine serum agar plus vancomycin from 9 of 11 patients. Sera from six animal species were tested to determine if any would support the growth of H. ducreyi. Horse and rabbit sera supported light growth of some strains. Fetal bovine serum supported good growth of all strains included in the study. Biochemical and physiological tests were done on the 17 isolates, a reference strain of H. ducreyi, and two reference strains of Haemophilus haemoglobinophilus. The results agreed with those reported by Kilian, except that H. ducreyi produced alpha-hemolysis in stabs on rabbit blood agar and was oxidase positive, three strains were urease positive, and CO2 improved the growth of seven strains. All 17 isolates were beta-lactamase positive. The reference strains were beta-lactamase negative.

The relationship between nine Haemophilus species and Haemophilus influenzae was studied by DNA-DNA hybridization, by transformation of H. influenzae to streptomycin resistance with heterospecific DNA, by competition of heterospecific DNA for transformation by homospecific DNA and by the lethal effect of heterospecific DNA on competent H. influenzae. H. parainfluenzae, H. parasuis, and H. aegyptius DNA transformed at more than 10% efficiency when compared to homologous transformation, but only H. aegyptius demonstrated, by hybridization, a relative binding ratio of more than 80%. H. aphrophilus and H. paraphrophilus DNA demonstrated a relative binding ratio of less than 30% and transformed H. influenzae at only 10(-5) the efficiency of homologous DNA, but they competed for H. influenzae transformation as well as or better than homospecific DNA. The data indicated that in some of the species sharing the common ecological habitat of the mammalian respiratory tract, sequences necessary for competition and efficient uptake into H. influenzae are present in large numbers in their DNAs, which nevertheless have little overall homology with H. influenzae DNA.

Fifty-seven clinical isolates of previously unidentified gram-positive, fermentative, nonsporeforming rods were studied and compared to the type strains of Oerskouia turbata and 0 . xanthineolytica. Thirty-five of the isolates were identified as Oerskouia species: 9 were identified as 0. turbata, and 26 were identified as 0. xanthineolytica. The Oerskouia cultures could be differentiated from the other isolates on the basis of the development of filamentous colonies. The genus Oerskouia, described by Prauser et al. (€9, Sukapure et al. (ll), and emended by Lechevalier (61, consists of yellow-pigmented organisms with branched hyphae. These hyphae break up into motile, rodlike elements that "appear bacterial" in smears. These rods are gram positive and non-acid fast. Previously described sources of the organisms are soil, aluminum hydroxide gel antacid, and dry grass cuttings (6). During the last 20 years, the Special Bacteriology Section (SBS), Bacteriology Division, Center for Disease Control (CDC), has received for identification a number of motile, gram-positive, nonsporeforming, yellow pigment-producing organisms isolated from clinical sources. These organisms were not recognized as belonging to any established species and were arbitrarily designated as group A Corynebacterium sp. Later, this group was divided into five subgroups, primarily on the basis of carbohydrate reactions. When these isolates were observed to have characteristics similar to those reported for the genus Oerskovia, a collaborative study between the Actinomycete Laboratory of CDC's Mycology Division and the SBS was initiated to further characterize and classify them. Listeria denitrificans was included in this study because two of our colleagues, D. Hollis and G. Wiggins, had noted during earlier work on the genus Listeria that many biochemical characteristics of this species were similar to those of the group A strains.