Biofilm formation in tracheal tubes, its bacterial content, and its interaction with ventilator gas flow were investigated. At least 50 mg (dry weight) of biofilm was found in 30 of 40 tracheal tubes used in intensive care patients for 2 h to 10 days. Electron microscopy showed bacteria in this layer, and quantitative studies showed that bacterial counts could reach up to 106/cm of tube length. Bacteria were cultured from the patient side of 18' of 78 heat and moisture exchanger-microbiological filter units removed from ventilator circuits. Particles were shown to detach from tracheal tube luminal biofilm and were projected up to 45 cm from the tracheal tube tip. Following contamination of the tracheal tube biofilm with a patient's own gastrointestinal flora, entrainment of bacteria in the inspiratory gas flow provides a mechanism for initial and repeated lung colonization.

Procalcitonin measurement potentially assists in the discrimination between severe lower respiratory tract infections of bacterial and 2009 H1N1 origin, although less effectively than CRP. Low values, particularly when combined with low CRP levels, suggested bacterial infection, alone or in combination with influenza, was unlikely.

AmpC b-lactamases (Bla AmpC ) are an emerging group of antimicrobial resistance determinants. The lack of an agreed Bla AmpC detection method hinders investigation of their epidemiology and understanding of their clinical significance. This study compared the sensitivity and specificity of phenotypic methods of Bla AmpC detection in a collection of 246 Enterobacteriaceae with a diverse range of b-lactam resistance profiles. The Bla AmpC screening methods evaluated were based on cephamycin, ceftazidime and cefepime susceptibility. These were compared with Bla AmpC screening using conventional ESBL detection methods. The confirmatory methods evaluated were biologically based assays, inhibitor-based assays, an AmpC Etest and a rapid chromogenic assay. A multiplex nucleic acid amplification test and the three-dimensional enzyme extraction assay were used as reference methods. Bla AmpC activity was present in 74 isolates. The majority of the enzymes were plasmid-encoded and belonged to the CMY, DHA and EBC families. The screening methods had sensitivities between 47 and 99 % and specificities of 45-95 %. The performance of confirmatory tests varied widely, ranging in sensitivity from 19 % to 97 % and in specificity from 88 % to 100 %. Only the Tris-EDTA and MAST ID D68C disc tests had a sensitivity and a specificity above 90 %. Further investigation is needed to establish the most suitable enzyme substrates, inhibitor types, inhibitor concentrations and interpretative cut-offs in order to refine the inhibitor-based methods. A simple disc-based protocol using cefoxitin non-susceptibility as a screening tool, followed by the Tris-EDTA method for confirmation, detects Bla AmpC activity with 95 % sensitivity and 98 % specificity.

Respiratory tract colonization with Scedosporium apiospermum in patients with chronic suppurative lung disease is a significant concern for lung transplantation candidates, since Scedosporium infections occurring posttransplantation are usually untreatable. Up to 10% of patients with cystic fibrosis attending our respiratory medicine unit have had Scedosporium organisms isolated from sputum samples. We therefore developed a molecular typing method to examine these isolates. Typing by PCR amplification of ribosomal intergenic spacer sequences demonstrated 20 different types from 52 isolates collected from the respiratory medicine unit and elsewhere in Australia. A single common type was isolated from 11 respiratory medicine unit inpatients. Two other types were isolated from more than one source: one from two respiratory medicine unit inpatients and one from two epidemiologically linked nonhuman sources. Multiple isolates were obtained from nine patients. This method demonstrated persistent carriage of isolates of the same type in one patient for 7 months. Two patients showed carriage of isolates with multiple typing patterns within a 3-month period. The high rate of isolation and the predominance of isolates with a single typing pattern from respiratory medicine unit patients may suggest transmission to patients from a source in the unit. There was no epidemiological evidence of direct patient-to-patient spread, and Scedosporium organisms were not isolated from dust, soil, or air samples from the unit. The source and route of transmission have yet to be determined.