A survey of all serogrouping schemata for Pseudomonas aeruginosa was conducted in order to identify all of the heat-stable major somatic antigens. In addition to the 12 antigens described in the schema of Habs (Z. Hyg. 144:218-228, 1957), five antigens from other schemata were found to be distinct. A grouping schema comprising 17 groups based on these antigens is proposed as the international serogrouping schema for P. aeruginosa. This schema is proposed as the backbone for future serotyping schemata that may include minor heat-stable antigens and heat-labile antigens. Several modifications of the schema are discussed. Variations of the schema can be adopted by individual laboratories without much confusion if the basic designations of the 17 antigens are retained. Pseudomonas aeruginosa was isolated byGessard (10) in 1882 as the cause of bluish green coloration of surgical dressings. Within 5 years of this discovery, the pathogenicity of this organism in experimental animals was established by Charrin (6). In 1896 Wasserman (37) differentiated antibodies directed against exotoxins and bactericidal antibodies directed against cells of P . aeruginosa, and in 1902 Achard et al.(1) described agglutination of P . aeruginosa by the serum of a man who was infected with the isolated agent. In 1903 Eisenberg (8) attempted to use such antisera to differentiate Bacillus pyocyaneus from Bacillus fluorescens. Similar work was reported by Jacobsthal (14) and Trommsdorf (32), who used a few strains of Pseudomonas species.In 1926 Aoki (3) divided 50 strains of P . aeruginosa into 22 serotypes. This work was followed by the work of Kanzaki (15), Sandiford (30), Mayr-Harting (21), Christie (7), and van den Ende (33). The P . aeruginosa strains used by these workers were not preserved, and later investigators formed their own serotyping schemata as needed. The use of different serotyping schemata and the failure to preserve reference strains for future studies made it impossible to compare epidemiological data.In 1957, Habs (11) described a serogrouping schema for P. aeruginosa that was based on 12heat-stable somatic antigens. This schema has been used by most European workers in serological studies of P . aeruginosa from human sources. In 1960, Sandvik (31) described a serological study of P . aeruginosa strains from bovine infections and reported eight serogroups among these strains. Veron (35) found that only one of the serogroups of Sanvik (31) represented a new antigen that was not present in the schema of Habs (11) and proposed that group I1 of Sanvik (31) be added to the schema of Habs (11) as group 13. In 1961, Verder and Evans (34) described an antigenic schema for P . aeruginosa that was based on both heat-stable and heat-labile antigens and divided the species into 10 serogroups on the basis of the heat-stable antigens. This schema has been used by many American investigators for clinical serotyping. Relationships between the serogroups in the schemata of Habs (11)

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The production of vascular permeability factor (PF) by certain strains of Pseudomonas aeruginosa has becen demonstrated in rabbits injected intradermally with culture filtrates followed by intravenous injection with Pontamine Sky Blue 6BX. The dose-response curve was found to be rectilinear when lesion diameters, within the range of 10 to 20 mm, were plotted against log dose. Thus, PF in test filtrates can be measured with reasonable accuracy by the concomitant testing of a reference PF. In contrast to the titers of PF obtained with Vibrio cholerae cultures, those with strains of P. aeruginosa were rather low. Thus far, PF has been demonstrated only in shallow still cultures of P. aeruginosa and not in shake cultures. A variety of commercial media were tested for the production of PF, but none was satisfactory. A synthetic medium that gave more reproducible and higher yields of PF was developed. Cultivation at 30 C generally gave higher yields of PF than at 37 C. PF was destroyed by heating at 60 C for 30 min or by digesting with trypsin or Pronase. Strains producing larger amounts of PF appeared to have greater virulence when inoculated onto the surface of burns in mice than those yielding little or no PF.

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