As part of the European Clostridium difficile infections (CDI) surveillance Network (ECDIS-Net), which aims to build capacity for CDI surveillance in Europe, we constructed a new network of hospital-based laboratories in Poland. We performed a survey in 13 randomly selected hospital-laboratories in different sites of the country to determine their annual CDI incidence rates from 2011 to 2013. Information on C. difficile laboratory diagnostic testing and indications for testing was also collected. Moreover, for 2012 and 2013 respectively, participating hospital-laboratories sent all consecutive isolates from CDI patients between February and March to the Anaerobe Laboratory in Warsaw for further molecular characterisation, including the detection of toxin-encoding genes and polymerase chain reaction (PCR)-ribotyping. Within the network, the mean annual hospital CDI incidence rates were 6.1, 8.6 and 9.6 CDI per 10,000 patient-days in 2011, 2012, and 2013 respectively. Six of the 13 laboratories tested specimens only on the request of a physician, five tested samples of antibiotic-associated diarrhoea or samples from patients who developed diarrhoea more than two days after admission (nosocomial diarrhoea), while two tested all submitted diarrhoeal faecal samples. Most laboratories (9/13) used tests to detect glutamate dehydrogenase and toxin A/B either separately or in combination. In the two periods of molecular surveillance, a total of 166 strains were characterised. Of these, 159 were toxigenic and the majority belonged to two PCR-ribotypes: 027 (n=99; 62%) and the closely related ribotype 176 (n=22; 14%). The annual frequency of PCR-ribotype 027 was not significantly different during the surveillance periods (62.9% in 2012; 61.8% in 2013). Our results indicate that CDIs caused by PCR-ribotype 027 predominate in Polish hospitals participating in the surveillance, with the closely related 176 ribotype being the second most common agent of infection.

Clostridium difficile is the cause of the nosocomial C. difficile infection (CDI). The conventional antibiotics used in CDI therapy are often unsuccessful, and recurrent infections may occur. Biofilm formation by C. difficile is associated with chronic or recurrent infections; biofilms may contribute to virulence and impaired antimicrobial efficacy. Manuka honey, derived from the Manuka tree (Leptospermum scoparium), is known to exhibit antimicrobial properties that are associated with its significant content of methylglyoxal, a natural antibiotic. The aim of the present study was to determine the antimicrobial effect of Manuka honey on clinical C. difficile strains belonging to four prominent polymerase chain reaction (PCR) ribotypes (RTs) (RT017, RT023, RT027 and RT046) and on their biofilm formation in vitro. Minimal inhibitory and bactericidal concentrations (MICs and MBCs, respectively) were determined using the broth dilution method. The biomass of the biofilm and the clearance of C. difficile biofilms by Manuka honey were determined using the crystal violet staining method. The MIC and MBC of Manuka honey for C. difficile strains were equal at 6.25% (v/v). PCR RT027 strains produced more biofilm in vitro than the other examined strains. Manuka honey effectively inhibited biofilm formation by C. difficile strains of different PCR RTs.

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