Acanthamoeba castellanii Neff supports the intracellular growth of Legionella pneumophila. When acanthamoebae were exposed to L. pneumophila for 1 h and then washed free of unassociated bacteria and placed in liquid culture, levels of viable amoeba-associated legionellae and legionellae free in the culture medium increased by three to four orders of magnitude in 48 to 72 h. However, most of the legionellae remained amoeba-associated and could be cultured only after disruption of the amoebae. Furthermore, legionella viability declined rapidly in amoeba culture medium alone or when bacteria and amoebae were separated by a microporous membrane. Therefore, direct amoeba-legionella contact is required for this growth. Infected acanthamoebae treated with cold acetone to permeabilize them to fluorescent-labeled anti-L. pneumophila antibody appeared to contain far more legionellae than amoebae fixed with glutaraldehyde so as to prevent antibody penetration. Electron micrographs of infected A. castellanii showed numerous bacteria, including some dividing forms, within vacuoles in the cytoplasm. These results together show that A. castellanii is able to provide an intracellular niche for the growth of L. pneumophila.

The rate of IE found in SAB is high when TOE is performed first line. There are no clear risk factors to improve yield or the type of echocardiography to be performed. Echocardiography should be performed in all cases and TOE should be considered where it is expected to influence management, as long as local resources allow.

BackgroundContamination of the inanimate environment around patients constitutes an important reservoir of MRSA. Here we describe the effect of introducing a universal disinfection wipe in all wards on the rates of MRSA acquisitions and bacteraemias across a large UK teaching hospital.MethodsA segmented Poisson regression model was used to detect any significant changes in the monthly numbers per 100,000 bed days of MRSA acquisitions and bacteraemias from April 2013 - December 2017 across QEHB.ResultsFrom April 2013 to April 2016, cleaning of ward areas and multi-use patient equipment by nursing staff consisted of a two-wipe system. Firstly, a detergent wipe was used, which was followed by a disinfection step using an alcohol wipe. In May 2016, QEHB discontinued the use of a two-wipe system for cleaning and changed to a one wipe system utilising a combined cleaning and disinfection wipe containing a quaternary ammonium compound. The segmented Poisson regression model demonstrated that the rate of MRSA acquisition/100,000 patient bed days was affected by the introduction of the new wiping regime (20.7 to 9.4 per 100,000 patient bed days; p <0.005).DiscussionUsing a Poisson model we demonstrated that the average hospital acquisition rate of MRSA/100,000 patient bed days reduced by 6.3% per month after the introduction of the new universal wipe.ConclusionWe suggest that using a simple one wipe system for nurse cleaning is an effective strategy to reduce the spread and incidence of healthcare associated MRSA.

BackgroundDiagnosis of C. difficile infection (CDI) is controversial because of the many laboratory methods available and their lack of ability to distinguish between carriage, mild or severe disease. Here we describe whether a low C. difficile toxin B nucleic acid amplification test (NAAT) cycle threshold (CT) can predict toxin EIA, CDI severity and mortality.MethodsA three-stage algorithm was employed for CDI testing, comprising a screening test for glutamate dehydrogenase (GDH), followed by a NAAT, then a toxin enzyme immunoassay (EIA). All diarrhoeal samples positive for GDH and NAAT between 2012 and 2016 were analysed. The performance of the NAAT CT value as a classifier of toxin EIA outcome was analysed using a ROC curve; patient mortality was compared to CTs and toxin EIA via linear regression models.ResultsA CT value ≤26 was associated with ≥72% toxin EIA positivity; applying a logistic regression model we demonstrated an association between low CT values and toxin EIA positivity. A CT value of ≤26 was significantly associated (p = 0.0262) with increased one month mortality, severe cases of CDI or failure of first line treatment. The ROC curve probabilities demonstrated a CT cut off value of 26.6.DiscussionsHere we demonstrate that a CT ≤26 indicates more severe CDI and is associated with higher mortality. Samples with a low CT value are often toxin EIA positive, questioning the need for this additional EIA test.ConclusionsA CT ≤26 could be used to assess the potential for severity of CDI and guide patient treatment.