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PubMed1985

Treatment of Pseudomonas and Serratia infections with ceftazidime.

The effectiveness and safety of parenterally administered ceftazidime were assessed in 20 patients with infections caused by Pseudomonas aeruginosa or Serratia marcescens. There were six infections involving the urinary tract, six wound infections, one respiratory tract infection, three septicaemias, one empyema, one mastoiditis and two infections of the epididymis. Sixteen of the isolates were resistant to at least one aminoglycoside antibiotic. Fifteen patients were clinically and bacteriologically cured, two patients improved, but had relapses and three patients showed no response. The lack of clinical response was due to the development of resistance by P. aeruginosa in two patients with osteomyelitis, one of whom also had diabetes mellitus. The third patient who had chronic lymphocytic leukaemia developed post-operative osteomyelitis and septicaemia with serratia. She did not respond to treatment. Side effects consisted of a positive direct Coomb's test without evidence of haemolysis in two patients and a mild maculopapular rash in one patient.

Adult↗

PubMed1985

Clinical efficacy of a synergistic combination of cefotaxime and amikacin against multiresistant Pseudomonas and Serratia infections.

The synergistic activity of cefotaxime and amikacin against 21 highly resistant Pseudomonas aeruginosa and Serratia marcescens isolates was evaluated in-vitro by the checkerboard tube dilution method and in-vivo in five patients with serious infections caused by these organisms. All isolates were resistant to gentamicin, tobramycin, amikacin, and cefotaxime. Synergy was observed in 90% of isolates and occurred when the MIC of amikacin was less than 256 mg/l and of cefotaxime less than 1024 mg/l. A clinical response occurred in 100% and bacteriological cure in 80% of patients. Our results demonstrate a high degree of synergism between amikacin and cefotaxime in-vitro and clinical efficacy in the treatment of infections due to multiply-resistant Pseudomonas and Serratia species.

Aged↗

PubMed1996

Analysis of the mechanism of the Serratia nuclease using site-directed mutagenesis.

Based on crystal structure analysis of the Serratia nuclease and a sequence alignment of six related nucleases, conserved amino acid residues that are located in proximity to the previously identified catalytic site residue His89 were selected for a mutagenesis study. Five out of 12 amino acid residues analyzed turned out to be of particular importance for the catalytic activity of the enzyme: Arg57, Arg87, His89, Asn119 and Glu127. Their replacement by alanine, for example, resulted in mutant proteins of very low activity, < 1% of the activity of the wild-type enzyme. Steady-state kinetic analysis of the mutant proteins demonstrates that some of these mutants are predominantly affected in their kcat, others in their Km. These results and the determination of the pH and metal ion dependence of selected mutant proteins were used for a tentative assignment for the function of these amino acid residues in the mechanism of phosphodiester bond cleavage by the Serratia nuclease.

Amino Acid Sequence↗

PubMed1997

Two genes encoding serine protease homologues in Serratia marcescens and characterization of their products in Escherichia coli.

A serine protease (SSP) of Serratia marcescens is one of the extracellular enzymes secreted from this Gram-negative bacterium. SSP is produced as a large precursor and converted to a mature protein by cleavages removing an NH2-terminal signal sequence and a COOH-terminal pro-region. This COOH-terminal pro-region is integrated into the outer membrane and has a functional role for the export of the mature protein across the outer membrane. Southern hybridization analysis with a DNA fragment encoding the COOH-terminal pro-region as the probe showed a wide distribution of nucleotide sequences encoding SSP exporter-like proteins among Serratia species. Moreover, S. marcescens IFO 3046, from which the ssp gene had been cloned, was found to contain two ssp homologues (ssp-h1 and ssp-h2). They were cloned and their nucleotide sequences were determined. The two ssp homologues were found to exist in tandem on the genome and their amino acid sequences showed 81% identity to each other. Both of them showed 55% identity in amino acid sequence to preproSSP. In addition, both showed end-to-end similarity to the 100 kDa serotype-specific antigen (Ssa1) of Pasteurella haemolytica. Escherichia coli JM105 containing ssp-h1 gene produced a 53 kDa protein corresponding to the NH2-terminal portion and a 49 kDa protein corresponding to the COOH-terminal portion, both of which were rigidly integrated in the outer membrane. Consistent with the significant similarity of the COOH-terminal portions of the homologues to that of SSP, they showed the ability to translocate the mature SSP part across the outer membrane into the medium. Furthermore, the NH2-terminal portion of the homologue was not translocated into the outer membrane without its COOH-terminal part. All of these data show that the SSP homologues are outer membrane proteins that are translocated into the outer membrane with the aid of the translocator function of their COOH-terminal part.

Amino Acid Sequence↗

PubMed1985

Follicle stimulating hormone binding inhibitor produced by the bacteria Serratia interacts with receptor for follicle stimulating hormone in calf testis membranes.

Bacteria of the genus Serratia, including a strain of Serratia liquifaciens isolated from contaminated porcine follicular fluid, produced an inhibitor of 125I-human follicle-stimulating hormone (hFSH) binding to calf testis membranes in vitro. In order to evaluate its potential usefulness and significance, we undertook studies to identify the site of action of this inhibitor. Large quantities (grams) of inhibitor-containing material (SL-1) were obtained by enrichment culture techniques and its chemical composition was determined. Follicle-stimulating hormone-binding inhibitory activity (FSH-BI) in SL-1 was associated with a protein-containing substance of approximately 30,000 Mr and also with larger Mr (greater than 300,000) forms. Preincubation studies demonstrated that binding inhibition by SL-1 was due to effects on the membranes rather than effects on the radioligand (125I-hFSH). Kinetics studies indicated that FSH inhibition by SL-1 was a relatively slow process that reached steady-state conditions between 23 and 25 h at 20 degrees C, in contrast to FSH, which reached steady-state conditions by 12 h at 20 degrees C. Estimates of FSH-BI activity, e.g., mass required to produce a 50% inhibition of 125I-hFSH binding, varied drastically when these kinetics differences were not taken into account. Our observations emphasize the need to establish steady-state conditions for each ligand before assessing mechanisms of action using Michaelis-Menton assumptions (e.g., competitive binding assays, Scatchard analyses).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1999

The phylogenetic position of Serratia, Buttiauxella and some other genera of the family Enterobacteriaceae.

The phylogenetic relationships of the type strains of 38 species from 15 genera of the family Enterobacteriaceae were investigated by comparative 16S rDNA analysis. Several sequences of strains from the genera Citrobacter, Erwinia, Pantoea, Proteus, Rahnella and Serratia, analysed in this study, have been analysed previously. However, as the sequences of this study differ slightly from the published ones, they were included in the analysis. Of the 23 enterobacterial genera included in an overview dendrogram of relatedness, members of the genera Xenorhabdus, Photorhabdus, Proteus and Plesiomonas were used as a root. The other genera formed two groups which could be separated, although not exclusively, by signature nucleotides at positions 590-649 and 600-638. Group A contains species of Brenneria, Buttiauxella, Citrobacter, Escherichia, Erwinia, Klebsiella, Pantoea, Pectobacterium and Salmonella. All seven type strains of Buttiauxella share 16S rDNA similarities greater than 99%. Group B embraces two phylogenetically separate Serratia clusters, a lineage containing Yersinia species, Rahnella aquatica, Ewingella americana, and also the highly related pair Hafnia alvei and Obesumbacterium proteus.

DNA, Ribosomal↗

PubMed1980

Phase variation in the genus Serratia.

During an investigation of the serotypes of Serratia marcescens present in a cancer centre, it was found that 8.7% of 241 strains had more than one H antigen. There were 19 diphasic and two triphasic strains. Antigen H1 was a component of the phases in the approximately half of the strains with multiple phases. This is the first report of such phases in the genus Serratia.

Antigens, Bacterial↗

PubMed2006

A generalized transducing phage (phiIF3) for the genomically sequenced Serratia marcescens strain Db11: a tool for functional genomics of an opportunistic human pathogen.

A bacteriophage (phiIF3) capable of mediating generalized transduction in Serratia marcescens strain Db11 has been isolated and characterized. The genome of this Serratia strain has recently been sequenced and is likely to become the reference strain for S. marcescens researchers. phiIF3 is most likely a virulent phage, which can transduce markers at frequencies of 10(-6) transductants per p.f.u. It has a lipopolysaccharide receptor and was determined to have a latent period of 50 min and a burst size of approximately 100 phages. The phage DNA was resistant to digestion with restriction enzymes. Electron microscopy showed phiIF3 to be a member of the family Myoviridae. This is the first report of a generalized transducing phage able to infect Db11 and this phage will be a valuable tool for functional genomic analysis of the pathogen host.

Bacteriophages↗

PubMed1982

An epidemiological study of Serratia marcescens infections by bacteriocin typing.

Bacteriocin sensitivity typing according to the method of Traub (Appl. Microbiol. 1971. 21: 837-840) was carried out on 226 clinical isolates of Serratia marcescens obtained from inpatients at Nagasaki University Hospital during the period from January 1976 to December 1978. The isolates were divided into 16 different bacteriocin types, mainly 26, 4, and 9. The distribution of the types suggests that Serratia marcescens infections may be caused by cross infection. Reproducibility of bacteriocin typing and the relationship between serotypes (O-antigen) and bacteriocin types are discussed in regard to the application of this method to the study of nosocomial infections.

Bacterial Infections↗

PubMed1990

Characterization of the type 1 fimbrial subunit gene (fimA) of Serratia marcescens.

The nucleotide sequence of a DNA fragment that contains the fimA gene, encoding the major fimbrial subunit, of Serratia marcescens IA506 and associated flanking sequences has been elucidated. In addition, the origin of transcription has been identified and is located 120 base pairs upstream of the fimA initiation codon. The predicted amino acid sequence of the FimA polypeptide exhibits some degree of sequence homology with the fimbrial subunits encoded by the fimA determinants of Klebsiella pneumoniae, Salmonella typhimurium, and Escherichia coli and also to Smf2, the major structural component of mannose-resistant (MR) fimbriae of S. marcescens. The Serratia adhesin that facilitates haemagglutination mediated by type 1 fimbriae is less susceptible to inhibition by D-mannose than has been observed to be the case in other type 1 fimbrial adhesins. The molecule conferring this adherence specificity has been shown to be distinct from the fimA gene product and, therefore, is analogous to the fimbrial systems reported in other species.

Amino Acid Sequence↗

PubMed1992

Expression of extracellular phospholipase from Serratia liquefaciens is growth-phase-dependent, catabolite-repressed and regulated by anaerobiosis.

Many members of the genus Serratia synthesize and excrete a number of extracellular hydrolytic enzymes. One of these is the phospholipase A1 from Serratia liquefaciens, the expression of which is growth-phase-dependent. Through the use of gene fusions and primer extension analysis we show that the expression of phospholipase is subject to positive transcriptional regulation of a dual promoter system; one promoter positioned approximately 600bp upstream from the phlA gene is responsible for the induction of phospholipase expression under anaerobic conditions, and the other promoter positioned 50bp upstream from the phlA gene is subject to catabolite repression and induced during the transition from exponential to late log-phase of bacterial growth. On the basis of sequence homology and behaviour in the relevant Escherichia coli mutants, we suggest that distant promoter to be Fnr-controlled and the proximal phlA promoter to be a member of the FIbB-controlled flagellar-chemotaxis regulon.

Amino Acid Sequence↗

PubMed1995

Induction of phospholipase- and flagellar synthesis in Serratia liquefaciens is controlled by expression of the flagellar master operon flhD.

When a liquid culture of Serratia spp. reaches the last part of the logarithmic phase of growth it induces the synthesis of several extracellular hydrolytic enzymes. In this communication we show that synthesis and secretion of the extracellular phospholipase is coupled to expression of flagella. Expression of flagella is demonstrated to follow a growth-phase-dependent pattern. Cloning, complementation studies and DNA-sequencing analysis has identified a genetic region in Serratia liquefaciens which exhibits extensive homology to the Escherichia coli flhD flagellar master operon. Interruption of the chromosomal flhD operon in S. liquefaciens results in non-flagellated and phospholipase-negative cells, but the synthesis of other exoenzymes is not affected. By placing the flhD operon under the control of a foreign inducible promoter we have shown that increased transcription through the flhD operon leads to induction of flagellar synthesis and phospholipase expression.

Amino Acid Sequence↗

PubMed1996

Involvement of N-acyl-L-hormoserine lactone autoinducers in controlling the multicellular behaviour of Serratia liquefaciens.

Several bacterial species possess the ability to differentiate into highly motile swarmer cells capable of rapid surface colonization. In Serratia liquefaciens, we demonstrate that initiation of swarmer-cell differentiation involves diffusible signal molecules that are released into the growth medium. Using high-performance liquid chromatography (HPLC), high resolution mass spectrometry and nuclear magnetic resonance (NMR) spectroscopy, we identified N-butanoyl-L-homoserine lactone (BHL) and N-hex anoyl-L-homoserine lactone (HHL) in cell-free Serratia culture supernatants. BHL and HHL are present in a ratio of approximately 10:1 and their structures were unequivocally confirmed by chemical synthesis. The swrl (swarmer initiation) gene, the predicted translation product of which exhibits substantial homology to the LuxI family of putative N-acyl homoserine lactone (AHL) synthases is responsible for directing synthesis of both BHL and HHL. In an swrl mutant, swarming motility is abolished but can be restored by the addition of an exogenous AHL. These results add swarming motility to the rapidly expanding list of phenotypes known to be controlled through quorum sensing.

4-Butyrolactone↗

PubMed2005

Biosynthesis of tripyrrole and beta-lactam secondary metabolites in Serratia: integration of quorum sensing with multiple new regulatory components in the control of prodigiosin and carbapenem antibiotic production.

Summary Serratia sp. ATCC 39006 (39006) uses a complex hierarchical regulatory network allowing multiple inputs to be assessed before genes involved in secondary metabolite biosynthesis are expressed. This taxonomically ill-defined Serratia sp. produces a carbapenem antibiotic (Car; a beta-lactam) and a red pigmented antibiotic, prodigiosin (Pig; a tripyrrole), which are controlled by the smaIR quorum sensing (QS) locus. SmaR is a repressor of Pig and Car when levels of N-acyl- l-homoserine lactones, produced by SmaI, are low. In this study, we demonstrate direct DNA binding of purified SmaR to the promoter of the Car biosynthetic genes and abolition of this binding by the QS ligand. We have also identified multiple new secondary metabolite regulators. QS controls production of secondary metabolites, at least in part, by modulating transcription of three genes encoding regulatory proteins, including a putative response regulator of the GacAS two-component signalling system family, a novel putative adenylate cyclase and Rap (regulator of antibiotic and pigment). Mutations in another gene encoding a novel predicted global regulator, pigP, are highly pleiotropic; PigP has a significant "master" regulatory role in 39006 where it controls the transcription of six other regulators. The PigP protein and its homologues define a new family of regulators and are predicted to bind DNA via a helix-turn-helix domain. There are regulatory overlaps between the QS and PigP regulons that enable the information from different physiological cues to be funnelled into the control of secondary metabolite production.

Amino Acid Sequence↗

PubMed1995

Serratia marcescens lung abscess in a child with autoimmune neutropenia.

Though Serratia marcescens is widely known to be the cause of serious infections in immunocompromised hosts, a lung abscess caused by S. marcescens is very rare. A 5 year old boy who had previously been diagnosed with autoimmune neutropenia was admitted because of fever and cough. In spite of treatment with some antibiotics, he developed a lung abscess. Aspiration of the pleural fluid revealed that S. marcescens was the pathogen of the disease. In the present case, there were feasible risk factors for the development of Serratia lung abscess namely neutropenia, chronic gingivitis at the time, and treatment with cyclosporin A. There are no reported cases of autoimmune neutropenia which developed into S. marcescens lung abscess in the literature as far as we can determine.

Autoimmune Diseases↗

PubMed1993

Stimulation and inhibition of polymorphonuclear leukocytes phagocytosis by lipoamino acids isolated from Serratia marcescens.

The role of the lipoamino acids (serratamolide and ornithine lipid), membrane lipid components of Serratia marcescens, was examined in phagocytosis and phagosome-lysosome fusion of human peripheral polymorphonuclear leukocytes. A mutant strain of Serratia marcescens (NS 38-09) lacking serratamolide was actively phagocytosed by human PMN, while the wild-type strain (NS 38) producing serratamolide was more resistant to phagocytosis by human PMN. Phagocytosis of killed Staphylococcus aureus coated with lipoamino acid (serratamolide), showed that they were more resistant to phagocytosis by PMN, while the cells coated with ornithine lipid or serratamic acid were phagocytosed more actively. Staphylococci coated with phosphatidylethanolamine or phosphatidylglycerol had no significant effect on phagocytosis by PMN. Phagosome-lysosome fusion by PMN labelled with acridine orange was examined by fluorescence microscopy. The fusion indices of lipoamino acid-coated staphylococci were the same as that of controls. Further, ornithine lipid-coated staphylococci stimulated the release of superoxide anion from PMN slightly, but serratamolide did not. These results suggested that serratamolide may contribute to the virulence of S. marcescens in vitro.

Depsipeptides↗

PubMed1996

Culture conditions affect cytotoxin production by Serratia marcescens.

Cytotoxins have been implicated in the pathogenesis of bacterial infections. In this study, the influence of different culture conditions was evaluated on cytotoxin production of Serratia marcescens. Parameters such as culture media, incubation temperature, starting pH of culture medium, aeration, anaerobiosis, carbon sources, iron concentration in he culture media, and release of cell-bond toxin by polymyxin B were investigated. The data suggest that this cytotoxin is predominantly extracellular and is not induced by iron limitation. Aerobic culture with shaking resulted in higher cytotoxicity than static aerobic or anaerobic culture. Bacteria grown in glucose, sucrose or galactose were more cytotoxic than those grown in inositol or maltose. The culture conditions that were identified as optimal for cytotoxin production by Serratia marcescens were incubation temperature ranging from 30 to 37 degrees C, in medium adjusted pH 8.5, with shaking. This work will contribute to further studies on the identification of this cytotoxic activity.

Bacterial Toxins↗

PubMed1998

Serratia marcescens and its extracellular nuclease.

Serratia marcescens produces an endonuclease with extraordinarily high specific activity that is released into the surrounding medium. This enzyme has been the focus of studies on gene regulation, protein secretion, endonuclease action, and protein structure; it has also been found to have many applications in biotechnology. Here we briefly review these different facets of research regarding the Serratia nuclease and summarize the current state of knowledge about this enzyme.

Bacterial Proteins↗