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The Yersinia HPI is present in Serratia liquefaciens isolated from meat.

AIMS: The aim of the study was to screen the Enterobacteriaceae flora of meat for the presence of bacteria harbouring the Yersinia high-pathogenicity island (HPI). METHODS AND RESULTS: Bacteria from 29 meat and 29 liver samples were isolated on violet-red bile glucose agar. A total of 197 isolates were screened for the presence of the irp2 gene, encoded within the HPI, by PCR. One isolate that was positive for irp2 gene was also positive for the fyuA, irp1, ybtP/ybtQ, ybtX/ybtS and int/asn tRNA genes by PCR. The presence of fyuA, irp1 and irp2 genes was confirmed by Southern hybridization. CONCLUSIONS: The isolate was identified as Serratia liquefaciens by sequencing of the 16S rRNA gene and by ribotyping. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of a Serratia harbouring the Yersinia HPI. Serratia is a frequently occurring Enterobacteriaceae genus in chill-stored meat.

Animals↗

[Contaminated infusions as cause of nosocomial Serratia marcescens septicaemia in children (author's transl)].

At the University Children's Clinic at Munich 25 cases of Serratia marescens septicaemia (mainly bacteriocin types 18 and 44) occurred within one year, predominantly in newborns and infants. Almost all of the children were seriously ill from an underlying illness or had been operated on. Two thirds had received antibiotics before the septicaemia occurred but they were ineffective. Of a total of nine antibiotics tested against 51 Serratia marcescens strains, nalidixic acid (99% sensitivity) and amikacin (100%) proved the most effective. Main source of the septicaemia were contaminated infusions, from which in as many as 35% of cases microorganisms, usually Serratia marcescens, had been isolated. Intensive hygienic measures at once terminated this "sepsis wave".

Age Factors↗

Application of oligonucleoside methylphosphonates in the studies on phosphodiester hydrolysis by Serratia endonuclease.

The endonuclease from Serratia marcescens is a non-specific enzyme that cleaves single and double stranded RNA and DNA. It accepts a phosphorylated pentanucleotide as a minimal substrate which is cleaved in the presence of Mg2+ at the second phosphodiester linkage. The present study is aimed at understanding the role of electrostatic and hydrogen bond interactions in phosphodiester hydrolysis. Towards this objective, six pentadeoxyadenylates with single stereoregular methylphosphonate substitution within this minimal substrate (2a-4b) were synthesized following a protocol described here. These modified oligonucleotides were used as substrates for the Serratia nuclease. The enzyme interaction studies revealed that the enzyme failed to hydrolyze any of the methylphosphonate analogues suggesting the importance of negative charge and/or hydrogen bond acceptors in binding and cleavage of its substrate. Based on these results and available site-directed mutagenesis as well as structural data, a model for nucleic acid binding by Serratia nuclease is proposed.

Binding Sites↗

In-vitro susceptibility of clinical isolates of Serratia species to antimicrobial agents.

The in-vitro activities of 12 antimicrobial agents against a total of 80 clinical isolates of Serratia marcescens and Serratia liquefaciens were determined by a broth microdilution method. Ampicillin and cefazolin were totally inactive against these organisms. The other beta-lactam antibiotics such as piperacillin, cefotaxime, ceftazidime, and the aminoglycosides such as gentamicin, tobramycin and netilmicin showed poor or moderate activity against Serratia isolates. Aztreonam and amikacin inhibited most of the strains tested. Imipenem and ciprofloxacin were very active in inhibiting all strains. Within the genus, S. liquefaciens was more resistant to aztreonam, ceftazidime and amikacin than S. marcescens.

Aminoglycosides↗

Bacteriocin typing of Serratia marcescens. A simplified system.

The authors describe a simplified system for the detection of bacteriocin production by Serratia marcescens with the use of six indicator strains, which include Escherichia coli, Klebsiella pneumoniae, Citrobacter diversus, Enterobacter aerogenes (two strains), and Serratia rubidaea grown on arabinose minimal medium plates. Of the 64 possible bacteriocin types, 11 were observed; 66% of the isolates tested were found to be one of three types. Occasionally more than one bacteriocin type was observed in an individual specimen; however, serotyping or antibiograms, or both, also indicated this was a different strain. The marcescin types were stable markers. With the use of this technic, different endemic strains of Serratia were shown to predominate in various areas of the hospital. In addition, when urinary tract isolates were compared with respiratory tract isolates, significant differences were found in the predominate types. The typing of these isolates by bacteriocin production was supported by serotype and antibiotype findings. The results suggest that this simple system may be a useful tool in a general hospital.

Bacteriocins↗

Specific and Nonspecific immunity to Serratia marcescens infection.

By specific active or passive immunization, mice were protected against lethal infection with Serratia marcescens. Animals that were immunized against organisms of the challenge strain O serotype survived, whereas animals that were immunized against other serratia O serotypes did not survive. Protective sera (from convalescent partients or immunized rabbits) contained a specific complement-independent opsonin. These sera also contained passive hemagglutinating and bacterial agglutinating antibodies in high titer. The protective and complement-independent opsonizing antibodies were associated with IgG fractions of the serum, while the agglutinating antibodies were primarily associated with the IgM. Mice also survived infection with Serratia after immunization against the Re595 Salmonella minnesota mutant. However, complement-independent opsonizing antibody was not demonstrated in these cross-protective sera.

Animals↗

The use of a DNA probe and PCR to examine the distribution of the aac(6')-Ic gene in Serratia marcescens and other gram-negative bacteria.

A nucleotide probe for the chromosomal aminoglycoside 6' acetyltransferase gene (aac(6')-Ic) of Serratia marcescens was used in non-radioactive dot-blot hybridization experiments with 186 strains belonging to ten different species of Serratia. The gene was only detected in strains of S. marcescens (all strains tested), and positive hybridization was seen irrespective of whether or not strains were kanamycin-resistant. An additional 180 strains belonging to 28 Gram-negative bacterial species other than in the genus Serratia did not hybridize with the probe. A rapid PCR test for the aac(6')-Ic gene was developed and used to confirm that the aac(6')-Ic gene is only found in S. marcescens. Southern hybridization analysis of S. marcescens chromosomal DNA demonstrated that the gene was usually located on a PvuII fragment with a putative isoleucine tRNA-2 gene, but polymorphisms with respect to the size of this fragment were observed.

Acetyltransferases↗

Multidrug-resistant Serratia marcescens bacteriuria related to urologic instrumentation.

An outbreak of urinary tract infections caused by multidrug-resistant Serratia marcescens lasted for 12 months and was found to be related to urologic instrumentation. Thirty-four patients had primary infections; four had cross-infections. Only six patients had indwelling bladder catheters. The median interval between instrumentation and initial isolation of Serratia was six days. Seventy-three percent of patients were symptomatic, two were bacteremic. No common instruments, personnel, or wards were identified, and environmental cultures failed to reveal the epidemic strain of Serratia. The outbreak ended when the instrument disinfectant was changed. Serotyping was identical in nine of ten isolates. Intraspecies conjugation demonstrated resistance transfer of gentamicin, tobramycin, carbenicillin, chloramphenicol, and co-trimoxazole. The enzyme 6'-N-acetyl transferase was responsible for gentamicin-inactivation in patient isolates and a transconjugate. Although no significant spread of this multidrug-resistance plasmid to other Enterobacteriaceae occurred in the hospital, two instances of apparent in vivo transfer to other bladder organisms occurred.

Adult↗

Products of defective lysogeny in Serratia marcescens SMG 38 and their activity against Escherichia coli and other Enterobacteria.

From a series of Serratia marcescens clinical isolates analysed with respect to bacteriocin production, one strain (SMG 38) was exceptional in that it produced two distinct phage-tail-like bacteriocins differing in morphology, sedimentation, heat sensitivity, and host range. The more active component (bc25) was effective against Serratia, while the other component (McG) inhibited growth of Escherichia coli, Salmonella typhimurium and Shigella sonnei, but not Serratia. Plaque formation on tested strains was negative except in the single case of the lysate of a subclone of SMG 38 which caused the production of a virulent phage, phi epsilon, in E. coli K12 RH 5108. This seems to be a rare event. Like the bacteriocin McG, phage phi epsilon used the same receptor protein, coded at about 30 min (locus fig) on the E. coli chromosome, as does the temperate and serologically unrelated phage phi gamma. Both McG and UV-irradiated phi epsilon killed sensitive bacteria. The survival rate depended on the input multiplicity and also on the indicator strain, and was increased by the presence of prophage phi 80 in the cell. When survivors were allowed to resume their growth under normal conditions, they showed cell elongation whatever their RecA phenotype. No difference was observed between the two agents with respect to these observations, except that McG, unlike irradiated phi epsilon, was inactive against Klebsiella pneumoniae UNF 5023, which possessed the Fig receptor.

Bacteriocins↗

Cloning and DNA sequence analysis of a bacteriocin gene of Serratia marcescens.

Serratia marcescens N28b synthesized and secreted a bacteriocin, with a molecular mass of 45 kDa, which was capable of inhibiting the growth of Escherichia coli. The expression of this bacteriocin was negligible unless induced with mitomycin C. The genes encoding the bacteriocin were cloned in plasmid pBR328. E. coli harbouring recombinant plasmid pBA189 or pBA289 expressed the Serratia marcescens N28b bacteriocin. The nucleotide sequence of the bss gene (Serratia marcescens N28b bacteriocin structural gene) was determined. The predicted amino acid sequence of the carboxy-terminal part of the bacteriocin 28b had a high degree of similarity to the pore-forming domains of colicins A, E1, B, N, Ia and Ib.

Amino Acid Sequence↗

Spore-forming Serratia marcescens subsp. sakuensis subsp. nov., isolated from a domestic wastewater treatment tank.

A strain (KREDT) that formed endospores and produced the pigment prodigiosin was isolated from activated sludge. The presence of spores in cells of strain KREDT was evident upon electron microscopy examination, heat treatment and the detection of dipicolinic acid in the cells. Biochemical characteristics, and 16S rDNA sequence and DNA-DNA homology data identified strain KREDT as Serratia marcescens. The major respiratory quinone of strain KREDT was found to be ubiquinone Q-8. The formation of endospores by Gram-negative bacteria has not been observed previously, and has never been reported in any species of Serratia. Here, it is shown that strain KREDT (JCM 11315T = CIP 107489T) represents a novel subspecies of S. marcescens, for which the name Serratia marcescens subsp. sakuensis is proposed.

DNA, Bacterial↗

Quorum-sensing-directed protein expression in Serratia proteamaculans B5a.

N-Acyl-L-homoserine-lactone-producing Serratia species are frequently encountered in spoiling foods of vegetable and protein origin. The role of quorum sensing in the food spoiling properties of these bacteria is currently being investigated. A set of luxR luxI homologous genes encoding a putative quorum sensor was identified in the N-(3-oxo-hexanoyl)-L-homoserine lactone (3-oxo-C6-HSL)-producing Serratia proteamaculans strain B5a. The 3-oxo-C6-HSL synthase SprI showed 79 % similarity with EsaI from Pantoea stewartii and the putative regulatory protein SprR was 86 % similar to the SpnR of Serratia marcescens. Proteome analysis suggested that the presence of at least 39 intracellular proteins was affected by the 3-oxo-C6-HSL-based quorum sensing system. The lipB-encoded secretion system was identified as one target gene of the quorum sensing system. LipB was required for the production of extracellular lipolytic and proteolytic activities, thus rendering the production of food-deterioration-relevant exoenzymes indirectly under the control of quorum sensing. Strain B5a caused quorum-sensing-controlled spoilage of milk. Furthermore, chitinolytic activity was controlled by quorum sensing. This control appeared to be direct and not mediated via LipB. The data presented here demonstrate that quorum-sensing-controlled exoenzymic activities affect food quality.

4-Butyrolactone↗

Biosynthesis of the red antibiotic, prodigiosin, in Serratia: identification of a novel 2-methyl-3-n-amyl-pyrrole (MAP) assembly pathway, definition of the terminal condensing enzyme, and implications for undecylprodigiosin biosynthesis in Streptomyces.

The biosynthetic pathway of the red-pigmented antibiotic, prodigiosin, produced by Serratia sp. is known to involve separate pathways for the production of the monopyrrole, 2-methyl-3-n-amyl-pyrrole (MAP) and the bipyrrole, 4-methoxy-2,2'-bipyrrole-5-carbaldehyde (MBC) which are then coupled in the final condensation step. We have previously reported the cloning, sequencing and heterologous expression of the pig cluster responsible for prodigiosin biosynthesis in two Serratia sp. In this article we report the creation of in-frame deletions or insertions in every biosynthetic gene in the cluster from Serratia sp. ATCC 39006. The biosynthetic intermediates accumulating in each mutant have been analysed by LC-MS, cross-feeding and genetic complementation studies. Based on these results we assign specific roles in the biosynthesis of MBC to the following Pig proteins: PigI, PigG, PigA, PigJ, PigH, PigM, PigF and PigN. We report a novel pathway for the biosynthesis of MAP, involving PigD, PigE and PigB. We also report a new chemical synthesis of MAP and one of its precursors, 3-acetyloctanal. Finally, we identify the condensing enzyme as PigC. We reassess the existing literature and discuss the significance of the results for the biosynthesis of undecylprodigiosin by the Red cluster in Streptomyces coelicolor A3(2).

Anti-Bacterial Agents↗

Isolation and functional analysis of luxS in Serratia plymuthica RVH1.

Autoinducer-2 (AI-2) is a signalling molecule proposed to act as a 'universal' signal for interspecies communication, regulating niche-specific genes with diverse functions in various bacteria. We have previously reported the production of AI-2 in strains from different Serratia species and recently, the luxS gene of two Serratia strains was isolated and shown to affect distinct phenotypes. In the current work, we have identified the luxS gene of Serratia plymuthica RVH1, and studied its effect on the N-acyl-L-homoserine lactone-regulated 2,3-butanediol fermentation, production of extracellular protease, chitinase, nuclease and antibacterial compound, and on biofilm formation. None of these phenotypes was affected by luxS knockout. However, overexpression of the luxS gene in RVH1, which was accompanied by an increased production of AI-2, resulted in a slower growth. This growth retardation was also observed when RVH1 was grown in spent culture medium from the AI-2-overproducing strain, but not in spent culture medium from the luxS knockout strain. These results suggest that luxS primarily fulfils a metabolic role and may not regulate a coordinated behaviour controlled by cell-to-cell signalling in S. plymuthica RVH1.

4-Butyrolactone↗

[Community acquired sepsis by Serratia rubidaea].

A 48-year-old male who had a past history of alcoholic pancreatitis and diabetes mellitus was admitted to our hospital due to chills and vomiting, on August 13, 1998. His body temperature was 38.0 degrees C, and he had the disturbance of consciousness, tachypnea, tachycardia and hepatomegaly with tenderness. Laboratory findings showed highly inflammatory reactions, DIC and hepatorenal dysfunction. Abdominal CT and US revealed multiple liver abscess with portal vein thrombus. Serratia rubidaea was detected in the blood culture. SBT/CPZ and TOB were administered and he recovered. This is a rare case of Serratia rubidaea sepsis. It is also necessary to pay attention to Serratia infections as well as S. marcescens.

Community-Acquired Infections↗

R factor-mediated antibiotic resistance in Serratia marcescens.

Nineteen of 39 multiresistant strains of Serratia marcescens isolated from clinical sources transferred antibiotic resistance to Escherichia coli or Klebsiella pneumoniae recipients. Marcesins and/or phage prevented effective resistance transfer to E. coli and attempts to select marcescin-resistant mutants of the E. coli recipient strain were unsuccessful. Transfer of resistance was demonstrated for all drugs tested except nalidixic acid. Approximately 90% of donors resistant to tobramycin, ampicillin, or carbenicillin transferred resistance to these drugs. High levels of transferred resistance (minimal inhibitory concentration, >2,500 mug/ml) were demonstrated particularly for ampicillin, carbenicillin, and kanamycin. Transmissibility of Serratia R factors was greatest between isogeneic strains of E. coli K-12. Comparative rates of spontaneous loss of R factor-mediated resistance indicated that Serratia R factors are less stable in E. coli and K. pneumoniae transcipients than in the indigenous hosts.

Bacteriocins↗

Sulfamethoxazole-trimethoprim-polymyxin therapy of serious multiply drug-resistant Serratia infections.

Nonpigmented multiply drug-resistant Serratia marcescens caused an extensive outbreak of infection at the Nashville Veterans Administration Hospital. Isolates were of one serotype resistant to all currently available antimicrobial agents for therapy of systemic infections except for occasional susceptibility to chloramphenicol and kanamycin. Frequently strains were susceptible to nalidixic acid, and all were susceptible to amikacin (BB-K8). Drug-resistant strains caused 130 infections, 12 bacteremias, and 7 infection-associated deaths. Combinations of antimicrobial agents were evaluated for synergism against Serratia strains from infected patients. "Checkerboard" isobolograms indicated in vitro static synergism between sulfamethoxazole, trimethoprim, and polymyxin (STP). Killing curves using clinically achievable concentrations of STP verified the bactericidal effect of STP against these strains. In a daily dosage of 1,600 mg of sulfamethoxazole and 320 mg of trimethoprim orally in combination with 100 to 300 mg of colistimethate parenterally, serum cidal levels at 1:8 or greater were achieved in five of six patients. Clinical improvement or microbiological cure was effected in four of six patients. STP may be potentially useful for selected Serratia infections for which single agents are unavailable or ineffective.

Clinical Trials as Topic↗

Differentiation of Klebsiella-Enterobacter (Aerobacter)-Serratia by biochemical tests and antibiotic susceptibility.

Studies were undertaken for the differentiation of members of the Klebsiella-Enterobacter (Aerobacter)-Serratia division by biochemical tests and antibiotic susceptibility. A total of 67 cultures were tested. Strain identification was readily accomplished with the use of motility tests and arabinose fermentation. In addition, a practical schema, based on sensitivity pattern, proved valuable in the classification of the different strains. Most (if not all) Klebsiella strains were susceptible to cephalothin, and all were sensitive to colistin. Enterobacter strains were resistant to cephalothin but susceptible to colistin. In contrast to those other members of the group, all Serratia strains were resistant to both cephalothin and colistin. The combination of a limited number of biochemical reactions and single-disc sensitivity tests appears to be a logical approach for the tentative identification of Klebsiella-Enterobacter (Aerobacter)-Serratia strains.

Bacteriological Techniques↗