Hemolytic activity of Serratia marcescens

Braun, Volkmar; Günther, Horst; Neuss, Burkhard; Tautz, C

doi:10.1007/bf00428852

Cited by 48 publications

(48 citation statements)

References 13 publications

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“…6 Recently Braun et a/. 7 noticed that Serratia marcescens strains rapidly lyse human erythrocytes in solution. On blood agar Serratia colanies show a narrow zone of a faint hemolysis.…”

Section: Lntroductionmentioning

confidence: 99%

The cell-bound hemolysin of Serratia marcescens contributes to uropathogenicity

Marre

Hacker

Braun

1989

Microbial Pathogenesis

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“…6 Recently Braun et a/. 7 noticed that Serratia marcescens strains rapidly lyse human erythrocytes in solution. On blood agar Serratia colanies show a narrow zone of a faint hemolysis.…”

Section: Lntroductionmentioning

confidence: 99%

The cell-bound hemolysin of Serratia marcescens contributes to uropathogenicity

Marre

Hacker

Braun

1989

Microbial Pathogenesis

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“…The strains we used were all strongly hemolytic in a liquid assay but caused only very narrow lysis zones on blood agar. Hemolysis was unrelated to the formation of the exoprotease (4). The hemolytic activity resided in the membrane fraction (4).…”

mentioning

confidence: 99%

Identification of the Serratia marcescens hemolysin determinant by cloning into Escherichia coli

Braun¹,

Neuss²,

Ruan³

et al. 1987

J Bacteriol

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A cosmid bank of Serratia marcescens was established from which DNA fragments were cloned into the plasmid pBR322, which conferred the chromosomally encoded hemolytic activity to Escherichia coli K-12. By transposon mutagenesis with TnlO00 and Tn5 ISSOL::phoA (TnphoA), the coding region was assigned to a DNA fragment, designated hly, comprising approximately 7 kilobases. Two proteins with molecular weights of 61,000 (61K protein) and 160,000 (160K protein) were expressed by the pBR322 derivatives and by a plasmid which contained the hly genes under the control of a phage T7 promoter and the T7 RNA polymerase. When strongly overexpressed the 160K protein was released by E. coli cells into the extraceliular medium concomitant with hemolytic activity. The genes encoding the 61K and the 160K proteins were transcribed in the same direction. Mutants We characterized an exoprotease of Serratia marcescens and studied its secretion (24). The strains we used were all strongly hemolytic in a liquid assay but caused only very narrow lysis zones on blood agar. Hemolysis was unrelated to the formation of the exoprotease (4). The hemolytic activity resided in the membrane fraction (4). Hemolytic bacteria have been shown to induce the release of the leukotrienes LTC4 and LTB4 from polymorphnuclear leukocytes, the release of histamine from rat mast cells, and chemiluminescence of neutrophils (2, 3). It was concluded that via these inflammatory mediators hemolysin may increase vascular permeability, edema formation, and granulocyte accumulation and thus contribute to the pathogenicity of Serratia spp. (18; W. Konig, Y. Faltin, J. Scheffer, B. Konig, H. Schoffler, and V. Braun, unpublished data).Hemolysis required actively metabolizing cells and could be inhibited by various energy poisons (4). Hemolytic activity was not found in the culture supernatant of various Serratia strains grown under different conditions, nor could it be released from cells (4). Therefore, we used a genetic approach to characterize the hemolytic determinant of S. marcescens. MATERIALS AND METHODSBacterial strains and culture conditions. The strains used are listed in Table 1. The rough mutant SN8 was obtained as a partially bacitracin-resistant derivative by using a method developed by H. Rotering (personal communication, this institute). Bacitracin affects 0-antigen synthesis by inhibiting recycling of the 0-antigen polyisoprenoid carrier (25).Colonies of S. marcescens W1436 that grew on 1/8 TY agar plates near a filter paper disk which contained 10 mg of bacitracin were examined by gel electrophoresis to determine whether they expressed smooth or rough lipopolysac-* Corresponding author.charide (27). Of 20 colonies tested, 4 lacked the ladder of bands characteristic of 0-antigen heterogeneity.Cells were routinely grown in TY medium (0.8% tryptone [Difco Laboratories], 0.5% yeast extract, 0.5% sodium chloride, pH 7). The antibiotic ampicillin (25 or 50 jig/ml) or kanamycin (50 jig/ml) was added to maintain the plasmids.Cosmid cloning. Chromosomal DNA o...

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“…This suggests that either the culture media contain sufficient calcium for haemolysis to take place or the observed haemolysis was calcium-independent , Calcium-independent haemolysins have been described for bacteria such as Proteus spp. and Serratia marcescens (Welch, 1987;Koronakis et al, 1987;Braun et al, 1985). It is possible that the haemolysin described herein may belong to the calciumindependent family of haemolysins and therefore be quite distinct from a-haemolysin.…”

Section: Discussionmentioning

confidence: 85%

Haemolysin production by strains of Verocytotoxin-producing Escherichia coli

Chart¹,

Jenkins²,

Smith³

et al. 1998

Microbiology

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Twenty-one strains of Verocytotoxin-producing Escherichia coli (VTEC) that hybridized with DNA probe CVD419 were examined for the ability to produce haemolysin. With solid media, all strains produced most haemolysin when grown in blood agar tubes and least when grown on blood agar plates incubated in air. Haemolysin production was increased considerably by incubating blood agar plates in an atmosphere comprising 8% carbon dioxide, 40% hydrogen and 52% nitrogen at 37 "C for 16 h, followed by 6 h at 21 "C in air. Haemolysin production was also increased when strains were grown on G agar containing the iron chelator ethylenediamine-di(olhydroxyphenylacetic acid) prior to subculture on blood agar. lntracellular haemolysin was detected in five out of the 21 strains of Em coli grown on L-agar in the atmosphere described above, but haemolysin was not detected in Gbroth culture supernatants. The haemolysins lysed guinea pig, mouse and ferret erythrocytes, but not human, rabbit, rat, turkey or chicken erythrocytes. Also, the addition of calcium ions to culture media was not required for haemolytic activity. It was concluded that haemolysins produced by VTEC appear to be quite distinct from E. coli a-haemolysin and resemble a form of phaemolysin.

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Hemolytic activity of Serratia marcescens

Cited by 48 publications

References 13 publications

The cell-bound hemolysin of Serratia marcescens contributes to uropathogenicity

The cell-bound hemolysin of Serratia marcescens contributes to uropathogenicity

Identification of the Serratia marcescens hemolysin determinant by cloning into Escherichia coli

Haemolysin production by strains of Verocytotoxin-producing Escherichia coli

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