A multidisciplinary approach has been adopted to investigate and identify the source of malodour in washing machines and the potential for crosscontamination of laundry. Four washing machines were olfactively graded, and the number of colony-forming units (CFUs) bacteria was determined in four specific locations. Then, samples of terry-towel and fleece were washed, without the use of detergent, in the machines, and the occurrence of malodour over a 52-h period was assessed. Analysis of the scrapings from the four locations in the two malodorous machines identified a plethora of volatile organic compounds (VOCs) by either olfactory detection or mass spectral identification post-gas chromatographic separation. In addition, microbiological analysis from the swabs from the four locations within all four washing machines was carried out. Quantitative analysis of VOCs from 66 microbiological isolates from either the washing machines or fabrics was carried out. In total, 10 VOCs were identified: dimethyl disulfide, 3-methyl-1-butanol, 2,4-dithiapentane, dimethyl trisulfide, 2-tridecanone, indole, 2-phenylethanol, isovaleric acid, isobutyric acid and 1-undecene.

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The molecular epidemiology of 66 NDM-producing isolates from 2 Pakistani hospitals was investigated, with their genetic relatedness determined using repetitive sequence-based PCR (Rep-PCR). PCR-based replicon typing and screening for antibiotic resistance genes encoding carbapenemases, other ␤-lactamases, and 16S methylases were also performed. Rep-PCR suggested a clonal spread of Enterobacter cloacae and Escherichia coli. A number of plasmid replicon types were identified, with the incompatibility A/C group (IncA/C) being the most common (78%). 16S methylase-encoding genes were coharbored in 81% of NDMproducing Enterobacteriaceae. With the worldwide spread of the NDM-1 gene and its variants (NDM-2 to NDM-8) (1, 2), molecular epidemiological studies of global isolates using various genotyping techniques are essential for gaining a better understanding of how this spread is occurring. India, Pakistan, and Bangladesh are clearly major reservoir countries for bla NDM , with numerous factors, such as antibiotic selection pressure, contributing to this current situation (3,4). This study examines a group of 66 NDM-1-producing isolates from Pakistan for their genetic relatedness, phylotype, plasmid replicon type, and plasmid transferability.All isolates were acquired from stool samples from 37 distinct patients at two military hospitals in Rawalpindi, Pakistan (5). The samples were collected from inpatients (35%) and outpatients (65%). The isolates were tested for susceptibility to 17 antimicrobials using the Vitek 2 system. The MICs for meropenem, doripenem, fosfomycin, and amdinocillin were determined using a standard agar dilution methodology (5).The isolates were reconfirmed for the presence of the carbapenem resistance gene bla NDM-1 by PCR, as previously described (6). PCR was also performed to detect bla , bla , bla VIM , bla IMP , bla KPC , bla CTX-M-15 , bla SHV , bla TEM , bla OXA-1 group, AmpC ␤-lactamases, bla CMY-2 , and the 16S rRNA methylase genes armA, rmtB, rmtC, and rmtF (6-9). The phylogenetic groups of Escherichia coli were determined using a multiplex (PCR)-based method (10).Repetitive sequenced-based PCR (Rep-PCR)-based typing by the DiversiLab system (bioMérieux, Oakleigh, Australia) was used for assessing clonal relatedness. A cluster of closely related isolates was defined as isolates sharing Ͼ95% similarity and indistinguishable isolates of Ͼ97% (11, 12). PCR-based replicon typing analysis (PBRT) was performed to determine the plasmid incompatibility (Inc) groups for all Enterobacteriaceae isolates (13).Ten genetically diverse E. coli isolates, based on different Rep-PCR profiles and phylogroups, were selected for transformation studies and typing by multilocus sequence typing (MLST). MLST

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