We developed and studied a molecular typing approach for Campylobacter spp. with restriction fragment length polymorphism (RFLP) analysis of the flagellin geneflaA in C. jejuni. Using polymerase chain reaction, we amplified theJfaA gene from strains comprising different HL:O serotypes by using a primer set directed at the conserved 5' and 3'flaA gene sequence to generate a 1.7-kb amplicon. The amplicon was further digested with the restriction enzyme DdeI, and the fragments generated were analyzed by agarose gel electrophoresis. In 43 non-outbreak strains of six common HL serotypes (HL 1, 2, 4, 5, 9, and 36) in the United States, 18 RFLP patterns were observed. In U.S. outbreak strains previously studied by 10 other typing methods, faA typing correlated with the HL serotype within each outbreak, and six additionalflaA types were identified. Our results suggest that RFLP analysis of theflaA gene from Campylobacter spp. has sufficient discrimination to be useful as a practical typing method for clinical and epidemiologic investigations.

Pittsburgh pneumonia agent (PPA) was recently cultivated from infected egg yold on charcoal yeast extract agar. PPA has now been isolated both from infected egg yolk and human lung tissue on charcoal yeast extract agar and on a new medium, buffered charcoal yeast extract agar. PPA resembles Legionella pneumophila and other Legionella-like organisms in requirements for growth and composition of fatty acids. It differs in genetic relatedness, antigenic composition, and colonial morphology and has distinctive characteristics that allow it to be identified. The name Legionella pittsburgensis species nova is proposed for this organism.

Campylobacter jejuni is the most common enteric pathogen isolated from university and college students in the United States. During the fall and winter quarters of the 1983-1984 academic year, the authors conducted a case-control study at the University of Georgia, Athens, Georgia, to identify risk factors for C. jejuni enteritis. Students with diarrhea whose cultures yielded C. jejuni were compared with controls matched by age, sex, and residence. A total of 45 case-control pairs were interviewed about exposures during the week before the case's onset of illness. The infections occurred sporadically and were caused by a wide variety of C. jejuni serotypes. Three risk factors were identified: eating fully cooked chicken, eating chicken reported to be raw or undercooked, and contact with a cat or kitten. No case reported drinking raw milk. No significant association was found between illness and the places where chicken meals were prepared or the specific manner in which chicken was cooked. Chicken may be the principal vehicle of transmission for sporadic Campylobacter enteritis among college students.

Pontiac fever, a unique epidemiologic form of legionellosis, is characterized by a short (one- to two-day) incubation period and a self-limited grippe-like illness without pneumonia. In 1968, the first documented outbreak of this syndrome affected persons who had entered a health department building in Pontiac, Michigan. Epidemiologic analyses clearly implicated as airborne agent and suggested that evaporative condenser water aerosols being disseminated by a defective air conditioning system played a key role in the outbreak. Guinea pigs that were exposed in the building and to laboratory aerosols of evaporative condenser water developed bronchopneumonia. Legionella pneumophilia (serogroup 1) was isolated from the exposed guinea pigs' lungs. Paired acute and convalescent serum specimens from 37 patients were tested by the indirect fluorescent antibody technique using L. pneumophila serogroup 1 antigen, and 31 (84%) had rises in titer from less than 32 to greater than or equal to 64.

Seventy-eight aerotolerant Campylobacter isolates were characterized phenotypically and by DNA hybridization (hydroxyapatite method at 50 and 65 degrees C). Two DNA relatedness groups were found. (i) Sixty-four strains belonged to aerotolerant Campylobacter DNA hybridization group 2. These organisms were isolated from humans, primarily with diarrheal illness, and animals on several continents. Strains were aerotolerant at 30 and 36 degrees C and catalase negative or weakly catalase positive, grew in media containing glycine and on MacConkey agar, were susceptible to nalidixic acid, and were resistant to cephalothin. The name Campylobacter butzleri sp. nov. is proposed for this group. (ii) DNA hybridization group 1 consisted of the type strain of Campylobacter cryaerophila and 13 additional strains isolated from 10 animals outside the United States and from three humans within the United States. This group was genetically diverse; five strains were closely related to the type strain of C. cryaerophila (DNA hybridization group 1A), and eight strains were more closely related to one another (DNA hybridization group 1B). Strains in DNA hybridization group 1B were phenotypically diverse, with two of eight strains resembling C. cryaerophila. The seven strains from DNA hybridization groups 1A and 1B which resembled C. cryaerophila and the C. cryaerophila type strain were aerotolerant only at 30 degrees C and catalase positive, did not grow in glycine or on MacConkey agar, were generally susceptible to nalidixic acid, and were resistant to cephalothin. The remaining six strains of DNA hybridization group 1B phenotypically resembled C. butzleri; however, they were generally catalase positive and susceptible to nalidixic acid and cephalothin. DNA hybridization group 1B is not designated as a separate species at this time since it cannot, with certainty, be separated genetically from C. cryaerophila or phenotypically from C. butzleri.

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We compared four phenotypic and six genotypic methods for distinguishing Campylobacter jejuni strains from animals and humans involved in four epidemics. Based on a comparison with epidemiologic data, the methods that correctly identified all strains in three milkborne outbreaks and one waterborne outbreak were heat-stable and heat-labile serotyping; multilocus enzyme electrophoresis (MEE); DNA restriction endonuclease analysis with BgIII, XhoI, PvuII, or PstI; and Southern blot and hybridization of PvuII-and PstI-digested DNA with Escherichia coli 16S and 23S rRNA (ribotyping). Biotyping, phage typing, plasmid analysis, and probing of BglII and XhoI DNA digests with C. jejuni 16S rRNA genes failed to correctly separate one or more strains. MEE, restriction endonuclease analysis, and ribotyping were the most sensitive methods and identified nine types among the 22 strains. These methods were also capable of further distinguishing strains within the same serotype. Data from MEE were also analyzed to calculate genetic relatedness among strains. Serotyping was the most discriminating phenotypic method, with eight and seven types distinguished by the heat-stable and heat-labile methods, respectively. MEE and ribotyping had several advantages over the other methods because they measure relatively stable and significant chromosomal differences and are applicable to other species and genera. These methods, however, are complex and not easily quantified; they are currently limited to specialized laboratories. When antisera are available, serotyping appears to be an effective and more practical approach to the identification of epidemic-related strains.

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