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[Infection with Serratia marcescens in newborn infants. Clinical aspects, therapy and disease course].

Until the 1950's Serratia marcescens was generally considered as non-pathogenic for humans. Since then the organism has been reported repeatedly as a cause of nosocomial infections. The major clinical concern about Serratia marcescens is its implication in epidemic infections and its resistance to usual antibiotics causing therapy to be difficult. We report the occurrence of Serratia marcescens as the cause of severe septicemia in three premature infants. Two infants showed a severe course of sepsis, the third infant suffered from additional meningitis and a brain abscess. All infants survived, but only one had no sequela.

Anti-Bacterial Agents↗

Serratia odorifera biogroup I: an emerging pathogen.

Gram-negative bacteremia is a common cause of infection in hospitalized patients. Serratia sepsis is known to cause clinically significant morbidity and mortality. The most common species involved is Serratia marcescens. Clinicians have been uncertain as to the role of Serratia odorifera biogroup 1 as a human pathogen because most isolates have not been associated with invasive disease. In previous publications, 12 cases have been described in which S odorifera biogroup 1 caused sepsis. These observations verify the organism's role as a human pathogen.

Aged↗

Pathobiology of amber disease, caused by Serratia Spp., in the New Zealand grass grub, Costelytra zealandica.

Amber disease in the New Zealand grass grub (Costelytra zealandica) is caused by some strains of Serratia entomophila or Serratia proteamaculans (Enterobacteriaceae). When treated with pathogenic isolates, larvae ceased feeding within 48 h, developed an amber coloration after 72 h, and entered a long chronic phase without feeding. An acute dose of 2-4 x 10(4) pathogenic bacteria was sufficient to produce disease in 50% of treated larvae. Time to death was directly related to temperature. At 15 degrees C, infected larvae remained in a chronic, nonfeeding state for more than 4 months prior to death. Nonpathogenic isolates, lacking the disease-causing plasmid (pADAP), had no effect on either feeding or disease. Twenty-four hours after ingestion, bacteria were found predominantly in the hindgut and growth occurred primarily within the fermentation chamber and in the head section of the larvae. Nonpathogenic strains did not multiply in treated larvae. Treatment of diseased larvae with antibiotic eliminated Serratia cells from the insects but did not result in restoration of feeding or the dark gut characteristic of the healthy larva.

Animals↗

Crystal structure of the 50 kDa metallo protease from Serratia marcescens.

The crystal structure of the 50 kDa metalloprotease from the Gram-negative bacterium Serratia marcescens has been solved and refined to a crystallographic R-factor of 0.192 at 1.80 A resolution. The structure is very similar to that of alkaline protease from Pseudomonas aeruginosa, in particular the calcium binding "parallel beta roll" motif is completely conserved. The N-terminal proteolytic domain shows the typical "metzincin" fold. The active sites of the two enzymes are slightly different, Tyr216 is a Zn ligand in the Serratia metallo protease. The loops 70-77 and 122-132, which encompass the active site cleft, differ due to insertions and deletions so that the Serratia metallo protease seems to have a more open site than the alkaline protease.

Amino Acid Sequence↗

Hemolysin as a marker for Serratia.

All Serratia marcescens strains (total of 33) of different sources were hemolytic including clinical strains previously classified as being nonhemolytic. DNA fragments of the two hemolysin genes hybridized with the chromosomal DNA of S. marcescens, S. liquefaciens, S. kiliensis, S. grimesii, S. proteamaculans, S. plymutica, S. rubridaea which were also hemolytic. The restriction pattern of the hemolysin locus differed in each strain. S. ficaria and S. marinorubra expressed a different hemolysin which was much smaller than the S. marcescens hemolysin since it diffused through dialysis membranes. The DNA of the latter strains did not hybridize with the S. marcescens hemolysin DNA probes. Some S. marcescens strains, S. kiliensis and S. liquefaciens also expressed in addition the small hemolysin. No hybridization was found with DNA of Escherichia coli, Salmonella typhimurium, Proteus mirabilis, Proteus vulgaris, Citrobacter freundii, Enterobacter cloacae, Klebsiella arerogenes, Klebsiella pneumoniae, Shigella dysenteriae, Yersinia enterocolitica, Yersinia pseudotuberculosus, Listeria sp., Aeromonas sp., Legionella sp. and a Meninococcus sp., indicating that the hemolysin DNA probes are specific for Serratia, or that the hemolysin genes occur rarely in genera other than Serratia.

Autoradiography↗

Hemolytic activity of Serratia marcescens.

A cell-bound hemolytic activity was found in several strains of Serratia marcescens. One Serratia cell per ten erythrocytes was sufficient to cause complete lysis of human erythrocytes within 2 h in the liquid assay. The hemolytic activity resided in the membrane fraction and could be inactivated by incubating cells with proteases. The hemolytic activity was greatly enhanced in actively metabolizing Serratia cells and was partially controlled by the iron supply. Hemolysis was accompanied by degradation of erythrocyte membrane proteins (band 3 and 6, glycophorin) and was independent of the blood group. The exoprotease secreted by S. marcescens in large amounts was not involved in hemolysis. Comparison with various hemolytic strains of Escherichia coli showed that hemolysis of erythrocytes was more pronounced with S. marcescens than with E. coli. In contrast to hemolysis by E. coli, lysis of erythrocytes by S. marcescens was not enhanced by Ca2+ ions.

Electrophoresis, Agar Gel↗

The metalloprotease gene of Serratia marcescens strain SM6.

Utilizing the DNA sequence of the metalloprotease from Serratia strain E-15, we isolated and sequenced the homologous gene from Serratia strain SM6. These two genes are similar at both the DNA and protein sequence level. Expression of the protease gene in Escherichia coli was achieved by use of the lac promoter. This resulted in the production and excretion of an immunologically detectable but inactive protein of slightly higher molecular weight than that from Serratia. We introduced the cloned gene into previously described protease mutants. The observed pattern of protease expression suggested that these mutations fall into three classes.

Amino Acid Sequence↗

Distribution and antibiotic susceptibility of extraintestinal clinical isolates of Klebsiella, Enterobacter and Serratia species.

A total of 451 extraintestinal, clinically relevant strains of the Klebsiella, Enterobacter and Serratia spp. isolated over a nine-month period from hospitalized patients at four different centers in Italy were investigated. Identification using the API 20E system showed that isolates belonged to 12 different species. Overall, strains of Klebsiella, Enterobacter and Serratia were in a ratio of approximately 3.4:2:1. Fifty-nine per cent of all strains were from urinary specimens, 12% from respiratory secretions, 10% from wounds and abscesses, and lower percentages from other sources. All strains were tested for their susceptibility to ten antibiotics. The rate of resistance to most drugs was generally greater in Enterobacter and Serratia than in Klebsiella. The overall incidence of strains of the intermediate category (i.e. between full sensitivity and resistance) was unexpectedly high. Both the relative frequency and the antibiotic susceptibility of strains of the various species varied from center to center; possible reasons for such differences are examined.

Drug Resistance, Microbial↗

Purification and properties of two chitinolytic enzymes of Serratia plymuthica HRO-C48.

The chitinolytic rhizobacterium Serratia plymuthica HRO-C48 was previously selected as a biocontrol agent of phytopathogenic fungi. One endochitinase (E.C. 3.2.1.14), CHIT60, and one N-acetyl-beta-1,4- D-hexosaminidase (E.C. 3.2.1.52), CHIT100, were purified and characterized. The endochitinase CHIT60, with an apparent molecular mass of 60.5 kDa, had a N-terminal amino acid sequence highly similar to that of chitinases A from Serratia liquefaciens and Serratia marcescens. The enzyme activity had its peak at 55 degrees C and pH 5.4, and increased by more than 20% in the presence of 10 mM Ca(2+), Co(2+) or Mn(2+). Activity was inhibited by 80% in the presence of 10 mM Cu(2+). CHIT100 appeared to be a monomeric enzyme with a molecular mass of 95.6 kDa and a pI of 6.8. Optimal activity was obtained at 43 degrees C and pH 6.6, and decreased by more than 90 % in the presence of 10 mM Co(2+) or Cu(2+). CHIT100 (100 microg ml(-1)) inhibited spore germination and germ tube elongation of the phytopathogenic fungus Botrytis cinerea by 28 % and 31.6 %, respectively. With CHIT60 (100 microg ml(-1)), the effect was more pronounced: 78 % inhibition of of germination and 63.9 % inhibition of germ tube elongation.

Amino Acid Sequence↗

Mass balance studies with 14C-labeled 2,4,6-trinitrotoluene (TNT) mediated by an Anaerobic desulfovibrio species and an Aerobic serratia species.

Investigations were carried out to evaluate the level of incorporation of radiolabeled 2,4,6-trinitrotoluene (TNT) and metabolites into the bacterial biomass of two different bacterial species after cometabolically mediated TNT transformation. Biotransformation experiments with 14C-TNT indicated that TNT was not mineralized; however, carbon derived from TNT became associated with the cells. It was found that more than 42% of the initially applied radiolabel was associated with the cell biomass after cometabolic 14C-TNT transformation with the strictly anerobic Desulfovibrio species strain SHV, whereas with the strictly aerobic Serratia plymuthica species strain B7, 32% of cell-associated 14C activity was measured. The remainder of the radiolabel was present in the supernatants of the liquid cultures in the form of different TNT metabolites. Under anoxic conditions with the Desulfovibrio species, TNT was ultimately transformed to 2,4,6-triaminotoluene (TAT) and both diaminonitrotoluene isomers, whereas under oxic conditions with the Serratia species, TNT was converted to hydroxylaminodinitrotoluenes and aminodinitrotoluenes, with 4-amino-2,6-dinitrotoluene (4ADNT) being the major end product. In both culture supernatants, small amounts of very polar, radiolabeled, but unidentified metabolites were detected. At the end of the experiments approximately 92% and 96% of the originally applied radioactivity was recovered in the studies with the Serratia and Desulfovibrio species, respectively.

Biodegradation, Environmental↗

Serratia keratitis transmitted by contaminated eyedroppers.

Serratia marcescens keratitis developed in three patients after keratoplasty. Two patients were using prednisolone sodium phosphate eyedrops and the third was using 0.5% timolol maleate eyedrops. All three cases resolved after treatment with topically and subconjunctivally administered antibiotics. Although S. marcescens was isolated from the outer grooves of the bottletops and from the inner surfaces of the eyedropper caps, it was not cultured from the solutions in the bottles. Moisture collecting in the dead space between the cap and bottle was apparently a culture medium for Serratia. When eyedrops were expressed into the patient's eyes, the eyes were inoculated with Serratia from the contaminated liquid flowing down the eyedropper shaft.

Aged↗

Availability of 6-hydroxynicotinic acid for rapid identification of Pseudomonas aeruginosa and Serratia marcescens.

Gas chromatography--mass spectrometry has been used to identify specific metabolites produced by Gram-negative bacteria such as Pseudomonas aeruginosa, Serratia marcescens, Klebsiella pneumoniae and Escherichia coli in a defined medium. 6-Hydroxynicotinic acid was detected in spent culture media of Pseudomonas aeruginosa and Serratia marcescens, but could not be detected in those of Klebsiella pneumoniae and Escherichia coli. The production of 6-hydroxynicotinic acid was recognized by the addition of nicotinic acid in urine with Pseudomonas aeruginosa or Serratia marcescens, but not without the addition of nicotinic acid. Among 10(5) Pseudomonas aeruginosa per 1 ml of urine (criteria for the diagnosis of urinary tract infection), 0.15 microgram of 6-hydroxynicotinic acid was detected in urine at 4 h incubation with nicotinic acid at the optimum pH of 6.9, 38 degrees C. The production of 6-hydroxynicotinic acid was proportional to the number of the bacteria and displayed a time dependency. These results suggest that the availability of 6-hydroxynicotinic acid might make for more rapid identification of bacteria than current methods.

Culture Media↗

Serratia bacteremia.

During a 6-yr period, 146 patients at our institution had Serratia bacteremia (3.8% of the total number of episodes of bacteremia), with an incidence of 1.24/1000 admitted patients. We chose a random group of 50 cases for clinical analysis in the present study. The disease was community-acquired in 8% of the cases and nosocomially-acquired in the remaining 92%. The bacteremia was unimicrobial in 84% and part of a polymicrobial bacteremia in 16% of the episodes. The most frequently isolated species of the Serratia genus was S. marcescens. Portals of entry, in decreasing order of frequency, were: urinary, unknown, respiratory, and surgical wound infections. Clinically, the most frequent finding was fever (100%). Shock occurred in 28% of the patients, and none of our cases showed evidence of disseminated intravascular coagulation. We found 62% of Serratia isolates resistant to gentamicin. Overall mortality was 38% and factors associated with a poor prognosis were: severity of the underlying disease, critical clinical situation at onset of bacteremia, presence in the intensive care unit (I.C.U.), occurrence of shock or polymicrobial bacteremia, portal of entry in the respiratory tract, and inadequate treatment.

Adolescent↗

Genotypic and phenotypic characterization of a biofilm-forming Serratia plymuthica isolate from a raw vegetable processing line.

Recently, we isolated from a raw vegetable processing line a Serratia strain with strong biofilm-forming capacity and which produced N-acyl-L-homoserine lactones (AHLs). Within the Enterobacteriaceae, strains of the genus Serratia are a frequent cause of human nosocomial infections; in addition, biofilm formation is often associated with persistent infections. In the current report, we describe the detailed characterization of the isolate using a variety of genotypic and phenotypic criteria. Although the strain was identified as Serratia plymuthica on the basis of its small subunit ribosomal RNA (16S rRNA) gene sequence, it differed from the S. plymuthica type strain in production of pigment and antibacterial compounds, and in AHL production profile. Nevertheless, the identification as S. plymuthica could be confirmed by gyrB phylogeny and DNA:DNA hybridization.

4-Butyrolactone↗

Genetic engineering, production and characterisation of monomeric variants of the dimeric Serratia marcescens endonuclease.

The Serratia nuclease is a non-specific endonuclease which cleaves single- and double-stranded RNA and DNA. It is a member of a large family of related endonucleases, most of which are dimers of identical subunits, with the notable exception of the Anabaena nuclease which is a monomer. In order to find out whether the dimer state of the Serratia nuclease is essential for its function we have produced variants of this nuclease which based on the crystal structure (Miller, M.D. and Krause, K.L. (1996), Protein Science 5, 24-33) were expected to be unable to dimerise. We demonstrate here that these variants, H184A, H184N, H184T and H184R, are monomers and have the same secondary structure, stability towards chemical denaturation and activity as the wild-type enzyme. This allows to conclude that the dimeric state is not essential for the catalytic function of the Serratia nuclease. In contrast, the S179C variant which is also a monomer shows little activity, presumably because this amino acid substitution changes the structure of the enzyme.

Bacterial Proteins↗

Meningoencephalitis due to serratia marcescens infection in neonates.

Six autopsy cases of Serratia meningoencephalitis were reported. The symptoms of this infection become apparent at an early stage in neonates; the meningoencephalitis in these six cases was severe, with or without inflammatory lesions in other organs. Serratia marcescens was originally described as a nonpathogenic organism; however, it is believed that after entering the blood stream of neonates, it becomes pathogenic, especially to the central nervous system. Meningoencephalitis due to Serratia marcescens is very severe and takes a rapid and progressive course against which no antibiotics have been found effective. Prevention of this infection is therefore considered of crucial importance.

Cerebrospinal Fluid↗

Strains of the genus Serratia as beneficial rhizobacteria of oilseed rape with antifungal properties.

Isolates of Serratia have been isolated from the rhizosphere of oilseed rape. The percentage of Serratia in this microenvironment was determined as 12.4% of the total antifungal bacteria. Serratia liquefaciens, S. plymuthica and S. rubidaea were found. All of the isolates showed an antifungal activity against different phytopathogenic fungi in vitro but the efficiency of strains was different. The antifungal mechanisms of 18 selected strains were investigated. Direct antifungal effect may be based on antibiosis (production of prodigiosin and pyrrolnitrin) and production of lytic enzymes (chitinases and beta-1,3-glucanases). Potent siderophores were secreted by the strains to improve the availability of iron. No strain was able to produce cyanide. Most of the strains secrete the plant growth hormone indole-acetic-acid which can directly promote the growth of roots. The mechanisms were specific for each isolate.

Acetic Acid↗

Biosynthesis of carbapenem antibiotic and prodigiosin pigment in Serratia is under quorum sensing control.

Serratia sp. ATCC 39006 produces the carbapenem antibiotic, carbapen-2-em-3-carboxylic acid and the red pigment, prodigiosin. We have previously reported the characterization of a gene, carR, controlling production of carbapenem in this strain. We now describe further characterization of the carR locus to locate the genes encoding carbapenem biosynthetic and resistance functions. A novel family of diverse proteins showing sequence similarity to the C-terminal domain of CarF (required for carbapenem resistance) is described. We also report the isolation of the locus involved in the biosynthesis of the red pigment, prodigiosin. A cosmid containing approximately 35 kb of the Serratia chromosome encodes synthesis of the pigment in the heterologous host, Erwinia carotovora, demonstrating, for the first time, that the complete prodigiosin biosynthetic gene cluster had been cloned and functionally expressed. We report the isolation of a third locus in Serratia, containing convergently transcribed genes, smaI and smaR, encoding LuxI and LuxR homologues respectively. SmaI directs the synthesis of N-acyl homoserine lactones involved in the quorum sensing process. We demonstrate that biosynthesis of the two secondary metabolites, carbapenem antibiotic and prodigiosin pigment, is under pheromone-mediated transcriptional regulation in this bacterium. Finally, we describe a new prodigiosin-based bioassay for detection of some N-acyl homoserine lactones.

Amino Acid Sequence↗