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Description of four novel species of Xenorhabdus, family Enterobacteriaceae: Xenorhabdus budapestensis sp. nov., Xenorhabdus ehlersii sp. nov., Xenorhabdus innexi sp. nov., and Xenorhabdus szentirmaii sp. nov.

The taxonomic affiliation was determined for four Xenorhabdus strains isolated from four Steinernema hosts from different countries. As compared to the five validly described Xenorhabdus species, i.e., X. nematophila, X. japonica, X. beddingii, X. bovienii and X. poinarii, these isolates represented novel species on the basis of 16S rRNA gene sequences and riboprint patterns, as well as by physiological and metabolic properties. They were named Xenorhabdus budapestensis sp. nov., type strain DSM 16342T, isolated from Steinernema bicornutam; Xenorhabdus ehlersii sp. nov., type strain DSM 16337T, isolated from Steinernema serratum; Xenorhabdus innexi sp. nov., type strain DSM 16336T isolated from Steinernema scapterisci; and Xenorhabdus szentirmaii sp. nov., type strain DSM 16338T, isolated from Steinernema rarum.

Animals↗

Phylogenetic analysis of enteric species of the family Enterobacteriaceae using the oriC-locus.

Phylogenetic analysis of 38 enteric species belonging to the Enterobacteraceae family was carried out using the non-coding locus oriC, the chromosomal replication origin. The oriC loci were amplified with conserved oligonucleotides and the PCR fragments were sequenced directly. The results establish a phylogenetic tree for the classification of different species of the genera Escherichia, Shigella, Salmonella, Enterobacter, Citrobacter, Klebsiella, Raoultella, Kluyvera, Cedecea and Buttiauxella. Functional important protein-binding sites located in oriC are well conserved throughout the enteric group. More over, due to a high overall divergence value phylogenetic trees were robust and well supported by bootstrap analysis. In comparison with 16S rDNA analysis, the oriC sequences indicated a greater evolutionary divergence for bacteria. We propose that the oriC locus might be a suitable phylogenetic marker for the identification and classification of bacteria, in particular for closely related species.

Bacterial Proteins↗

Enzymatic characterization of TEM-63, a TEM-type extended spectrum beta-lactamase expressed in three different genera of Enterobacteriaceae from South Africa.

The extended-spectrum beta-lactamase, TEM-63, was identified in three separate genera of South African isolates: Proteus mirabilis, Klebsiella pneumoniae, and Escherichia coli. This paper describes identification of the gene in these isolates and compares relative rates of hyrolysis between TEM-63 and other known ceftazidimases.

Amino Acid Substitution↗

Epidemiology of aerobactin production in Enterobacteriaceae.

Clinical isolates (496) of 10 different enterobacterial genera were studied for aerobactin excretion, colicin production and antibiotic resistance. In the case of Escherichia coli, the incidence of aerobactin-positive strains in 108 blood isolates (45%) was not significantly different from that corresponding to faecal sources (41%). Although colicin V production was much more frequently associated with aerobactin production than other colicins, colicin V was only produced by 26% of aerobactin excreters. Some strains were aerobactin-negative and colicin V producers. Aerobactin production seemed to be significantly associated with plasmidic antibiotic resistance. The production of the siderophore is described for the first time in Proteus, Serratia and Morganella strains.

Blood↗

Electrochemical measurement of trace concentrations of biological hydrogen produced by Enterobacteriaceae.

Accurate measurements of the hydrogen gas produced by Escherichia coli and Hafnia alvei pure cultures during glucose metabolism were performed under different growth conditions: stagnant, with magnetic stirring or with vibrational shaking. These measurements were carried out using an electrochemical hydrogen sensor based on a platinum-coated solid polymer electrolyte membrane (Pt-SPE). The results obtained were dependent on the hydrodynamic conditions of the growth, with greater hydrogen production being associated with the stagnant conditions. These measurements will eventually enable us to elucidate whether the pathway used for glucose metabolism is either strictly or mainly anaerobic and to modify experimental conditions so as to influence the reaction.

Culture Media↗

Antimicrobial susceptibility of inducible AmpC beta-lactamase-producing Enterobacteriaceae from the Meropenem Yearly Susceptibility Test Information Collection (MYSTIC) Programme, Europe 1997-2000.

Among 11345 clinical isolates from 31 European centres in the MYSTIC (Meropenem Yearly Susceptibility Test Information Collection) Programme (1997-2000), the potential AmpC-producing pathogens were Enterobacter spp. (904 cases), Citrobacter spp. (144) and Serratia marcescens (288). Resistance to ceftazidime or cefotaxime (11-34%) and piperacillin/tazobactam (12%) was observed, indicating stably derepressed expression of AmpC cephalosporinases. Meropenem (MIC(90), 0.25 mg/l; >99% susceptible) and imipenem (MIC(90), 2 mg/l; 98% susceptible) were active, even against these stably derepressed AmpC-producers: Enterobacter spp. (305 strains), Citrobacter spp. (29) and S. marcescens (35). The overall rank order of spectrum (% susceptible) versus the stably derepressed subset of isolates was: meropenem=imipenem (98%)>cefepime (89%)>gentamicin (73%)>ciprofloxacin (62%)>tobramycin (60%) >piperacillin/tazobactam (21%). We suggest increased attention in Europe to: (1) the recognition of these resistant phenotypes, (2) infection control practices, and (3) limiting the overuse of certain selecting extended-spectrum beta-lactam agents (e.g. ceftazidime).

Anti-Bacterial Agents↗

Analysis of plasmids mediating ANT (2") and AAC(3)-II based gentamicin resistance in clinical isolates of Enterobacteriaceae.

The genetics of ANT(2") and AAC(3)-II mediated gentamicin resistance in Gram-negative bacteria obtained from Hacettepe University Hospital in Turkey, was studied. The plasmid DNA analyzed by agarose gel electrophoresis revealed a heterogeneous plasmid population. Conjugal transfer appeared to be an efficient mechanism for the dissemination of resistance to gentamicin/tobramycin/kanamycin/streptomycin/chloramphenicol/cephalothin , which was the common resistance pattern in the population. Plasmids of 46, 60, 70 and 80 Mdal in size were found to be conjugative. Among these a 70 Mdal plasmid was isolated more frequently; it was harboured by the strains exhibiting either of the enzyme profiles, ANT(2") or AAC(3)-II. Like the substrate profiles of the donors, their enzymatic activities were also maintained in the respective transconjugants. Based on the donor and transconjugant resistance markers, agarose gel electrophoresis and transfer properties, ANT(2") and AAC(3)-II dependent gentamicin resistance was found to be plasmid-linked. However, the evidence of a possible chromosomal location for the gentamicin resistance suggested that the resistance might well be of a transposable nature.

Anti-Bacterial Agents↗

Laboratory transmission of enterobacteriaceae by the oriental cockroach, Blatta orientalis.

Methods of maintaining and feeding and of infecting cockroaches with pathogenic organisms were investigated.Cockroaches fed on known concentrations of Escherichia coli O119, Esch. coli O1, Alkalescens Dispar O group 2 and Shigella dysenteriae 1 (Shiga's Bacillus) were maintained in Petri dishes. The effect of various diets on the survival of these organisms within the hind-gut and faeces was observed. With a ;normal' diet of gruel Esch. coli O119 was isolated for up to 20 days, Esch. coli O1 for 17 days and ADO 2 for 15 days. Sh. dysenteriae 1 was isolated only sporadically to the third day.

Animals↗

Occurrence of multiple antibiotic resistance and R plasmids in Enterobacteriaceae isolated from children in the Sudan.

The prevalence of resistance to six commonly-used antimicrobial agents in faecal coliforms from children in Khartoum, Sudan was studied. A relatively high prevalence of resistance was found, ranging from 96% of children with isolates resistant to ampicillin to 70% of children with isolates resistant to chloramphenicol. Seventy-seven percent of children had isolates with high-level resistance to trimethoprim (MIC greater than 1000 micrograms/ml). Twenty-nine different resistance patterns were found. Thirty-nine percent of the children had isolates resistant to all six antibiotics studied, and 80% of children had isolates resistant to at least four. Transfer of resistance to each of the antimicrobials, in varying combinations, was demonstrated, but did not occur for all resistance patterns. Plasmid analysis showed plasmids ranging from 160 MDa to 2.8 MDa and isolates contained from one to five plasmids of different sizes. There were no consistent relationships between resistance pattern and plasmid profile, but multiple resistance transfer was mediated commonly by plasmids with a molecular weight of 62 MDa. The high prevalence of potentially transferable antibiotic resistance in gut commensals of children in the Sudan may be of importance in the management of enteric and other infections requiring antimicrobial treatment.

Ampicillin Resistance↗