Mahmoud A. Halablab

Legionella drozanskii sp. nov., Legionella rowbothamii sp. nov. and Legionella fallonii sp. nov. : three unusual new Legionella species

Seven strains of Legionella-like amoebal pathogens (LLAPs) were characterized on the basis of their cultural and staining characteristics, biochemical reactions, serology, cellular fatty acids (CFAs), isoprenoid quinone composition, total DNA relatedness, analysis of 16S rRNA and macrophage infectivity potentiator (mip) gene sequence analyses. All seven strains exhibited limited growth on buffered charcoal yeast extract alpha (BCYE) agar, required cysteine for growth and contained branched-chain CFAs and quinones typical of Legionella species. The bacilli were Gram-negative and catalase-positive. There were varying degrees of serological cross-reactions between these LLAP strains and other previously described Legionella species. Results from the various tests revealed that four LLAP strains represent three unusual new species of Legionella: Legionella drozanskii sp. nov., type strain LLAP-1T; Legionella rowbothamii sp. nov., type strain LLAP-6T; and Legionella fallonii sp. nov., type strain LLAP-10T. Three other LLAP strains, designated LLAP-7FL, LLAP-7NF and LLAP-9, were shown to be members of the species Legionella lytica. The deductions made from the phenetic characteristics of these bacteria were consistent with the phylogenetic relationships inferred from 16S rRNA and mip gene sequence analyses. This study is the first to speciate LLAP strains on the basis of data including quantitative DNA hybridization.

The pattern and kinetics of substrate metabolism of Campylobacter jejuni and Campylobacter coli.

Aims:  The main aim was to investigate the patterns and kinetics of substrate oxidation by Campylobacter jejuni and C. coli.

Distribution of Bacteria on Hands and the Effectiveness of Brief and Thorough Decontamination Procedures Using Non-medicated Soap

Our perception of the role of hand washing in the clinical situation is based on experimental studies in which test-bacteria are usually inoculated onto the skin surface and removed using hand washing preparations containing antiseptics. In this study, we have investigated the distribution of bacteria on the hands of volunteers and the effectiveness of long (3 minute) and brief (10 second) washes in removing both naturally-occurring and artificially-inoculated bacteria (Micrococcus sp.), using only soap and water. There was a tenfold reduction in median counts of artificially inoculated bacteria following both long and brief washes. However, less than 50% of naturally-occurring bacteria were removed and, for hands previously disinfected by immersion in 70% ethanol, the washing procedure increased bacterial counts. In both unwashed hands, and hands washed following a strict protocol, the mean variation in counts of naturally-occurring bacteria at different sites (wrists, dorsal surface, palmar surface, fingertips and interdigital spaces) was only two-fold. The efficiency of recovery of naturally-occurring organisms was estimated by repeated swabbing, to be more than 60%. The data question the value of typical hand wash procedures recommended by many authorities for use in clinical situations and of the perfunctory hand washes frequently adopted by nursing staff in busy wards. Experimental evidence is required to justify procedures and to identify the precise circumstances in which they are of value.

An in situ method for determining bacterial survival on food preparation surfaces using a redox dye

A simple method is described for the direct enumeration of viable bacteria dried on test surfaces. Inoculated surfaces were overlayed with agar and after incubation nitroblue tetrazolium solution (pale yellow) was used to stain colonies (purple) at the agar‐test surface interface. Stained colonies could be readily detected and counted even against the opaque background of ceramic tile or stainless steel or when present within opaque films of milk or serum. Recovery of bacteria by this method was approximately fivefold greater than using a conventional swabbing procedure. The method was used to demonstrate the marked effect of the composition of the suspension fluid, in which bacteria were dried, and the length of surface exposure upon bacterial survival.

A novel liposome-based electrochemical biosensor for the detection of haemolytic microorganisms

A biosensor strategy for rapid amperometric detection of organisms was investigated in which a redox mediator was entrapped within liposomes. The selective release of mediator by haemolytic bacteria, followed by signal generation, confers differentiation between haemolytic and non-haemolytic species. The potential of this approach is illustrated by results for various strains of Listeria monocytogenes, Listeria welshimeri and Escherichia coli.

Cholesterol Protects Acholeplasma laidlawii Against Oxidative Damage Caused by Hydrogen Peroxide

The aim of this study was to determine whether cholesterol, added to the cell growth medium or to cell suspension buffer, could protect Acholeplasma laidlawii cells against the toxic effects of hydrogen peroxide (H2O2). Variable concentrations of cholesterol (0.05–1.0 mg/ml) were added to the A.laidlawii suspension buffer and to the growth medium. Cells were then washed carefully and incubated with 0.001% (v/v) H2O2 at 37°C for 30 min and the viability was determined. The results indicated that cells were more viable in the presence of cholesterol than were cells grown in the absence of cholesterol. In addition, the oxygen uptake rate resulting from the oxidation of 5.5 mmol/L glucose was 2-fold and 4-fold higher for cells grown in medium supplemented with 0.05 and 0.50 mg/ml cholesterol, respectively, compared to cells grown in a medium with no added cholesterol. These findings indicate that cholesterol might play a role in protecting Mollicutes against the oxidative damage caused by H2O2.

Alternative to fluorescence assays to monitor fusion between Acholeplasma laidlawii cells and liposomes

Aims: To develop a new technique as an alternative to the fluorescence assays and electron microscopy for the purpose of monitoring the cell‐liposome fusion.