The Genus Serratia

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236 F. Grimont and P.A.D. Grimont CHAPTER 3.3.11 Table 10. Correlation between biogroups and serotypes in Serratia marcescens. Biogroup a Serotypes included A1 O5:H2; O5:H3; O5:H13; O5:H23; O10:H6; and O10:H13 A2/6 O5:H23; O6,14:H2; O6,14:H3; O6,14:H8; O6,14:H9; O6,14:H10; O6,14:H13; O8:H3; and O13:H5 A3 O3:H5; O3:H11; O4:H5; O4:H18; O5:H6; O5:H15; O6,14:H5; O6,14:H6; O6,14:H16; O6,14:H20; O9:H9; O9:H11; O9:H15; O9:H17; O12:H5; O12:H9; O12:H10; O12:H11; O12:H15; O12:H16; O12:H17; O12:H18; O12:H20; O12:H26; O13:H11; O13:H17; O15:H3; O15:H5; O15:H8; O15:H9; O17:H4; O18:H21; O18:H26; O22:H11; and O23:H19 A4 O1:H1; O1:H4; O2:H1; O2:H8; O3:H1; O4:H1; O4:H4; O5:H1; O5:H6; O5:H8; O5:H24; O9:H1; O13:H1; O13:H11; and O13:H13 A5/8 O2:H4; O3:H12; O3,21:H12; O4:H12; O5:H4; O6,14:H4; O6,14:H12; O8:H4; O8:H12; O15:H12; and O21:H12 TCT O1:H7; O2:H7; O4:H7; O5:H7; O5:H19; O7:H7; O7:H23; O10:H8; O10:H9; O11:H4; O13:H7; O13:H12; O16:H19; O18:H9; O18:H16; O18:H19; O19:H14; O19:H19; O20:H12; and O24:H6 a Biogroup A1 is composed of biotypes A1a and A1b; A2/6 is composed of A2a, A2b, and A6; A3 of A3a, A3b, A3c, and A3d; A4 of A4a and A4b; A5/8 of A5, A8a, A8b, and A8c; and TCT of TCT and TT. used to achieve final identification of H antigens. The following absorbed sera need to be prepared: anti-H:8 absorbed by H:10; anti-H:10 absorbed by H:8; and anti-H:3 absorbed by H:10. Other cross-reactions are overcome by dilution of sera. The immobilization test works because H antigens are monophasic in S. marcescens. A total of 175 complete (O+H) serotypes have been identified at this time. No plasmid or phage has yet been found to alter the serotype of a strain. Some striking correlations between antigenic composition and biotype have been shown (Grimont et al., 1979a). Pigmented biotypes and nonpigmented biotypes are antigenically segregated. Serotyping subdivides the biotypes, but when two biotypes correspond to the same serotype, these biotypes usually differ by only one biochemical reaction. Biotypes can be united into “biogroups” in order to clarify biotype-serotype correlation (Table 10). These biogroups may be valid infraspecific taxons. Serotyping of Other Species An antigenic scheme for S. ficaria is presently available (Grimont and Deval, 1982). Four somatic (O) and one flagellar (H) antigens were identified that defined four serovars. All American strains studied (isolated from the fig wasp or from a human patient) belonged to serotype O1:H1; strains from the Mediterranean region (Sicily, Tunisia, France) were not so antigenically uniform and all four serotypes were found in figs from Sicily. An antigenic scheme for S. rubidaea has also been devised (Grimont et al., manuscript in preparation). Nine somatic and five flagellar antigens were identified. A total of 53 strains were distributed among 14 complete (O+H) serotypes. Bacteriocin Typing of Serratia marcescens Two different schemes have been developed for typing Serratia marcescens strains by susceptibility to bacteriocins (“marcescins”). The system of Traub (1980) was developed since 1971 (Traub et al., 1971) and uses marcescins (incomplete phage structures) produced by 10 selected strains. In 1980, 73 bacteriocin types had been described and about 93% of strains were typable (Traub, 1980). In this system, bacteriocin type 18 is often associated with serotype 06/14:H12 in multipledrug-resistant nosocomial strains. Farmer (1972b) independently developed another system of typing based on susceptibility to marcescins. After mitomycin induction, 12 bacteriocin-producing strains were selected, and the 12 marcescins could subdivide 93 strains into 79 types. Typing of S. marcescens by production of bacteriocin was also developed by Farmer (1972a). Twenty-three strains susceptible to bacteriocins were selected to serve as indicator strains. Each unknown S. marcescens strain is treated with mitomycin, and the marcescin produced (if any) is identified according to the pattern of susceptibility demonstrated by indicator strains. This method could be used to type 91% of the strains studied. Phage Typing of Serratia Farmer (1975) has developed a phage typing system common to Serratia marscens, S. liquefaciens, and S. rubidaea (marinorubra). With 74 phages, 95% of the S. marcescens strains and 50% of the S. liquefaciens and S. rubidaea strains could be typed. Phage typing was found to be convenient for subdividing prevalent serotypes (Negut et al., 1975). F. Grimont (1977) independently developed another phage typing system
CHAPTER 3.3.11 The Genus Serratia 237 for nosocomial strains isolated in Bordeaux. Most of these S. marcescens strains gave phage susceptibility patterns that corresponded to phage types described by F. Grimont (1977); however, strains isolated in other cities of France were less often typable, and strains received from other countries were rarely typable in this system. Typing by Enzyme Electrophoresis By using agar gel electrophoresis, Grimont et al. 1977a and Grimont and Grimont (1978c) detected seven different proteinases (called P 5, P 6, P 7, P 9a, P 9b, P 11, and P 12) produced by strains of S. marcescens. Each S. marcescens strain can produce one to four proteinases; 651 strains of S. marcescens, isolated over a period of six years in a French hospital, gave 33 different proteinase patterns (called zymotypes). However, six zymotypes alone accounted for 76% of the isolates. Proteinase electrophoresis was thus used as an epidemiological marker (Grimont and Grimont, 1978c). Goullet (1978, 1981) showed that the Serratia species were characterized by distinct electrophoretic patterns of their esterases. However, these esterase profiles were not used as epidemiological marker. Enzyme polymorphism among 99 S. marcescens isolates was determined by electrophoresis of nine enzymes (Gargallo-Viola, 1989). More than one electromorph was found for seven of these enzymes (alanine dehydrogenase, catalase, NADP-dependent malic enzyme, 6-phosphogluconate dehydrogenase, glucose-6-phosphate dehydrogenase, NADP-dependent glu- tamate dehydrogenase, and indophenol oxidase), and 33 distinctive electrophoretic types. One group was represented exclusively by isolates belonging to nonpigmented biotypes recovered almost entirely (97%) from clinical samples. The other group comprised all isolates belonging to a pigmented biotype. Electrophoresis of glucose-6phosphate dehydrogenase alone can distinguish S. marcescens strains belonging to a pigmented biotype from those belonging to a nonpigmented biotype (Gargallo et al., 1987). A very good correlation was found between electrophoretic periplasmic protein patterns, enzyme electrophoresis, and biotyping by carbon source utilization (Gargallo-Viola and Lopez, 1990). Restriction Patterns Electrophoretic patterns of fragments produced after cleavage of total DNA by restriction endonucleases have been used to compare S. marcescens strains in epidemiological studies (McGeer et al., 1990). A new, generally applicable typing method has recently been proposed by Grimont and Grimont (1986) in which the DNA restriction fragments carrying rRNA genes are visualized after hybridization with a labelled E. coli 16+23S rRNA probe (rRNA gene restriction patterns). Five to seven DNA restriction fragments (after digestion by Bam HI) or 11 to 13 fragments (after digestion by EcoRI) were observed with all Serratia strains tested (Grimont and Grimont, manuscript in preparation). The different S. marcescens biogroups display different rRNA gene restriction pattern. This method should be useful in resolving occasionally uncertain (or conflicting) results given by serotyping and biotyping. Literature Cited Ackerman, L. J., Kishimoto, R. A., Emerson, J. S. 1971. Nonpigmented Serratia marcescens arthritis in a Teju (Tupinambis tequixin). American Journal of Veterinary Research 32:823–826. Alford, L. R., Holmes, N. E., Scott, W. T., Vickery, J. R. 1950. Studies on the preservation of shell eggs. Australian Journal of Applied Science 1:208–214. Altemeier, W. A., Culbertson, W. R., Fullen, W. D., McDonough, J. J. 1969. Serratia marcescens septicemia. A new threat in surgery. Archives of Surgery 99:232–238. Barnum, D. A., Thackeray, E. L., Fish, N. A. 1958. An outbreak of mastitis caused by Serratia marcescens. Canadian Journal of Comparative and Medical Veterinary Science 22:392–395. Bascomb, S., Lapage, S. P., Willcox, W. R., Curtis, M. A. 1971. Numerical classification of the tribe Klebsielleae. Journal of General Microbiology 66:279–295. Bergan, T., Grimont, P. A. D., Grimont, F. 1983. Fatty acids of Serratia determined by gas chromatography. Current Microbiology 8:7–11. Bergey, D. H., Harrison, F. C., Breed, R. S., Hammer, B. W., Huntoon, F. M. 1923. Bergey’s manual of determinative bacteriology. Baltimore, Williams & Wilkins. Berkowitz, D. M., Lee, W. S. 1973. A selective medium for isolation and identification of Serratia marcescens. Abstracts of the Annual Meeting of the American Society for Microbiology 1973:105. Bizio, B. 1823. Lettera di Bartolomeo Bizio al chiarissimo canonico Angelo Bellani sopra il fenomeno della polenta porporina. Biblioteca Italiana o sia Giornale di Letteratura Scienze a Arti 30:275–295. Boam, G. W., Sanger, V. L., Cowan, D. F., Vaughan, D. P. 1970. Subcutaneous abscesses in iguanid lizards. Journal of the American Veterinary Medical Association 157:617–619. Bollet, C., Gulian, C., Mallet, M. N., Estrangin, E., de Micco, P. 1989. Isolation of Serratia rubidaea (Stapp) from fresh and spoiled coconuts. Tropical Agriculture 66:342–345. Bouvet, O. M. M., Grimont, P. A. D., Richard, C., Aldová, E., Hausner, O., Gabrhelová, M. 1985. Budvicia aquatica gen. nov., sp. nov.: a hydrogen sulfide-producing member of the Enterobacteriaceae International Journal of Systematic Bacteriology. 35:60–64.
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