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Method: Microbiological reports from all sputum samples sent from patients at the AWACFC 2013–2016 inclusive were retrospectively reviewed for the reporting of S.m. Sex, age and FEV1% predicted of patients culturing S.m on consecutive years were recorded and an annual FEV1% decline calculated for the years post S.m isolation. Patients other microbiological growths were recorded. Results: In 2013 18 of 300 (6%) patients cultured S.m at least once in the year. This rose to 23 from 294 (7.8%) in 2014, 26 from 292 (8.9%)in 2015 and 31 of 292 (10.6%) in 2016.18 (10 male) patients isolated S.m at least once on three or more consecutive years. Their mean age was 30.2 years and mean FEV1% 56.4% (range 25–103% predicted) (AWACFC overall mean FEV1 69.2%). Six of these 18 (33%) had previous isolates of non tuberculous mycobacterium (NTM) (overall AWACFC NTM prevalence 7%), 11 (61%) had previous isolates of aspergillus and 11 (61%) had no Pseudomonas aeruginosa (PsA) growths (total AWACFC PsA prevalence 51%). The mean annual FEV1% decline of this group of 18 patients with recurrent growths of S.m is 2.5% pa. Conclusion: It would appear that the prevalence of S.m in our CF cohort is increasing and may reflect changes in antimicrobial prescribing. The patients isolating S. m recurrently have a wide range of FEV1% severity, but they have an increased prevalence of aspergillus (61% VS our centre prevalence 17%) and NTM growths but a reduced prevalence of PsA compared to our centre average. Their FEV1% decline is slightly above our median decline. Further studies are needed to fully describe the longterm effects on S.m colonisation. 138 Use of rpoB sequence clustering for species-level identification of the Elizabethkingia genus from UK CF patients, and an investigation of associated antimicrobial susceptibility D. Kenna1, A. Fuller1, K. Martin1, R. Pike1, C. Perry1, J. Shah1, D. Lilley1, R. Hill1, N. Woodford1, J. Logan1, J. Turton1. 1Public Health England, Antimicrobial Resistance and Healthcare Associated Infections (AMRHAI) Reference Unit, London, United Kingdom Objectives: The Elizabethkingia genus comprises E. meningoseptica, E. miricola and E. anophelis. Since 2013, submissions of Elizabethkingia spp. to the UK CF reference laboratory have increased. MALDI-ToF analysis has been a popular method for identifying these organisms in hospitals, but the current standard Bruker database does not include E. anophelis, which results in misidentification of some isolates. We developed an rpoB sequencing method for species identification within this genus and compared the results with those from MALDI-ToF analysis and urease testing. Methods: Forty-four isolates from 38 patients identified as belonging to the Elizabethkingia genus by MALDI-ToF were analysed by rpoB sequencing. An E. miricola-specific ureG PCR was compared with urea hydrolysis on agar slopes. MICs of a panel of antibiotics were determined for twelve isolates. Results: rpoB sequencing separated E. meningoseptica, E. miricola and E. anophelis type/reference strains into three discrete clusters, and revealed 43/44 (99%) CF isolates were E. miricola; the exception clustering with only 98.9% similarity to E. miricola. MALDI-ToF analysis could not definitively distinguish between E. miricola/E. meningoseptica. All 44 isolates were ureG PCR-positive, confirming rpoB clustering results, yet only 7/22 isolates tested hydrolysed urea, suggesting interpretation was problematic. The 12 isolates tested were resistant to most antibiotics, with full susceptibility to minocycline and co-trimoxazole only. Conclusion: rpoB sequencing highlighted both the prevalence of E. mircola among UK CF patients, and inaccuracies in commonly used identification methods in these multi-resistant organisms. 139 Prevalence of Aspergillus fumigatus and clinical interaction with Pseudomonas aeruginosa in the Dutch CF population L. Slabbers1, P. Merkus2. 1Radboud Univerversity, Pediatrics, Nijmegen, Netherlands, 2Radboud Univerversity, Nijmegen, Netherlands Objectives: Literature suggests that under in vitro conditions, PA produces more elastase in the presence of AF. An enhanced decline of FEV1 was observed in a young cohort of co-infected CF patients. The aim of this study was to investigate whether a negative interaction exists between co-

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infection with PA and AF on lung function in the Dutch CF population compared with infection with either, after adjustment for confounders. Methods: A National Dutch Filamentous fungal biobank with sputum cultures of patients treated at the 7 Dutch CF centers was established to study the prevalence of fungi in a 3 year period. Data from the National CF registry were used to assess the relationship between FEV1% of predicted of patients with positive sputum samples of PA with and without positive samples for AF, with adjustment for the following confounders: Age, BMI, severity of class mutations, CFRD, ABPA, azithromycin and inhaled antibiotic use. Results: Chronic PA infection was observed in 475 (34.5%) of the Dutch CF patients, of which 259 (54.5%) had a positive AF culture. AF was found in 605 (44.0%) of the entire Dutch CF patients of whom 259 (42.8%) were chronically infected with PA. A multivariate analysis showed that there is no difference in FEV1% of predicted in CF patients infected with only PA, AF or co-infected patients. Conclusion: We conclude from this large scale National study that, afterappropriate correcting for relevant confounders, CF patients with AF infections without PA have on average a higher FEV1% of predicted compared to patients with PA with and without co-infections of AF. These findings have relevant clinical consequences because they suggest that there is no reason to start antifungal treatment as a routine measure when AF is cultured in CF patients, even in the presence of a co-infection with PA. 140 Co-colonisation of the cystic fibrosis airways with A. fumigatus and P. aeruginosa is associated with poorer health: an Irish registry analysis E. Reece1, J. Renwick1, R. Segurado2, S. M. Clean3, P. Greally4. 1Department of Clinical Microbiology, Trinity College Dublin, Trinity Centre for Health Science, Tallaght hospital, Dublin, Ireland; 2CSTAR, School of Public Health, Physiotherapy and Population Science, Woodview House, University College Dublin, Dublin, Ireland; 3School of Biomolecular and Biomedical Sciences, University College Dublin, Dublin, Ireland; 4Department of Respiratory Medicine, The National Children’s Hospital, Tallaght Hospital, Dublin, Ireland Objectives: A. fumigatus and P. aeruginosa are the most common fungal and bacterial pathogens isolated from the cystic fibrosis (CF) airway, respectively. Our aim was to determine the effect of different colonisation patterns of these two pathogens on the clinical status of patients with CF. Methods: A retrospective analysis of data from the Cystic Fibrosis Registry of Ireland was performed to determine the effect of intermittent and persistent colonisation with A. fumigatus or P. aeruginosa or co-colonisation with both microorganisms on clinical outcome measures (FEV1, number of hospitalisations, respiratory exacerbations and antimicrobials prescribed, and complications of CF, including CFRD and ABPA) in patients with CF. Results: The prevalence of A. fumigatus and P. aeruginosa colonisation was 11% (5% persistent, 6% intermittent) and 31% (19% persistent, 12% intermittent) in the Irish CF population, respectively. Co-colonisation with both pathogens was associated with a 13.8% reduction in FEV1 ( p = 0.011), higher numbers of exacerbations ( p = 0.042), hospitalisations ( p = 0.023) and antimicrobials ( p = 0.014) compared to non-colonised patients and these clinical outcomes were comparable to those of patients persistently colonised with P. aeruginosa. Intermittent and persistent A. fumigatus colonisation was not associated with poorer clinical outcomes. Conclusion: CF patients co-colonised with A. fumigatus and P. aeruginosa had similarly poor clinical outcomes to those persistently colonised with P. aeruginosa, emphasising the clinical significance of co-colonisation with these microorganisms. 141 Aspergillus fumigatus degrades the respiratory mucins MUC5B and MUC5AC A. Cowley1, A. Horsley2, D. Denning3, D. Thornton1. 1University of Manchester, Wellcome Trust Centre for Cell-Matrix Research, Manchester, United Kingdom; 2Manchester Adult CF Centre, Manchester, United Kingdom; 3 University of Manchester, Institute of Inflammation and Repair, Manchester, United Kingdom Objectives: Aspergillus fumigatus (Af ) is the most common fungal pathogen in CF airways. As the first point of contact for Af spores following inhalation,

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the mucus barrier provides an important site for host-pathogen interactions. The details of these interactions, and the effects of Af on mucus barrier properties, are poorly understood. We have therefore investigated the effects of Af on the major structural components of airway mucus, the polymeric gel-forming mucins MUC5AC and MUC5B. Methods: To study the effects of Af on mucin size and glycosylation, purified mucins were treated with Af culture filtrates and analysed using agarose gel electrophoresis, rate-zonal centrifugation, and lectin assays. Protease assays were used to determine whether different classes of proteases were involved in degrading mucins. Size exclusion chromatography was used to fractionate the Af filtrate, and mass spectrometry of active fractions applied to identify mucin-degrading enzymes. To identify potential mucin cleavage sites, N- and C-terminal domains of MUC5B were expressed in mammalian cells, and the effects of Af filtrates on these recombinant proteins were studied. Results: Af was able to degrade airway mucins, targeting both protein and carbohydrate regions of the molecules. Lectin reactivity fell for both mucins over 24hrs, and protein immune-reactivity was undetectable after 2–4hrs of treatment. Mucin degradation was protease-dependent and two specific proteases were isolated: alkaline serine protease 1 and neutral protease 2. Both the N- and C-terminal domains of MUC5B were targeted by Af proteases, with the N-terminus appearing more susceptible to proteolytic cleavage. Conclusions: A. fumigatus is able to degrade both protein and carbohydrate components of airway mucins. This may have important pathophysiological consequences, including use of mucins as a nutrient source or as a way of altering mucus barrier properties to promote airway colonisation. 142 Characteristics of fungal pathogens detected in patients from a single CF center U. Krivec1, A. Kotnik Pirs1, M. Praprotnik1, M. Aldeco1, D. Lepej1, A. Zver1, R. Tomazin2, T. Matos2. 1University Childrens’ Hospital Ljubljana, University Medical Centre Ljubljana, Unit for pulmonary diseases, Ljubljana, Slovenia; 2 Institute of Microbiology and Immunology, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia Objectives: Fungal pathogens are often detected in respiratory samples from patients with cystic fibrosis (CF). Even though often considered harmless, persistent colonization/infection has been associated with worse respiratory outcomes. Aim: To assess the prevalence and characteristics of fungal pathogens detected in respiratory samples from a single pediatric CF center. Methods: All mycological respiratory samples’ detections from CF patients in a 5 year period were reviewed. Analysis focused on sample characteristics and prevalence of isolated pathogens. Results: The study included samples from 70 patients, median age 14 yrs, rage 3–25 yrs. A total of 1202 respiratory samples were collected during 2012–2016. There were 628 (52.2%) sputa, 444 (36.9%) deep pharyngeal aspirates, 68 (5.7%) bronchoalveolar lavage fluids and 61 (5.2%) other samples. 1013 (84.3%) samples were positive for fungi. There were 1411 fungal isolates in total belonging to 19 different genera. The most prevalent species were: Candida albicans (58.3%) and Aspergillus fumigatus (23.3%). Among other fungal species associated with CF there were: Clavispora lusitaniae (2.4%), Exophiala dermatitidis (1.3%), Rasamsonia argillacea (0.4%). Scedosporium apiospermum/Pseudallescheria boydii species complex was not detected in this patient population. Conclusions: The biodiversity of isolated fungi in CF patients is high with representatives from 19 different genera. Like in other European countries, C. albicans and A. fumigatus, together representing more than 80% of all isolated fungi. CF associated emerging fungal pathogens like E. dermatitidis and R. argillacea were detected, whereas S. apiospermum/P. boydii species complex was not present in this group of CF patients. 143 Fungal epidemiology and diversity over the past four years in adult cystic fibrosis patients attending the All Wales Adult CF Centre (AWACFC) L. Speight1, R. Dhillon2, L. White2, D. Thomas3, R.I. Ketchell1, D. Lau1, J. Duckers1. 1University Hospital Llandough, All Wales Adult CF Centre,

Penarth, United Kingdom; 2Public Health Wales, Department of Microbiology, University Hospital of Wales, Cardiff, United Kingdom; 3Public Health Wales, Communicable Disease Surveillance Centre, Cardiff, United Kingdom Background: Over the past few years there has been increasing interest in the fungal burden in CF respiratory samples. There was a local clinical perception that the prevalence of fungi recovered from the CF cohort was increasing. Aim: To give a retrospective overview of the diversity of fungi cultured from the sputum samples of adult CF patients attending the AWACFC from 2013 to 2016 and determine whether the perceived increase in burden was accurate. Method: Microbiological reports from all sputum samples sent from patients at the AWACFC from 2013 to 2016 inclusive were retrospectively reviewed for the reporting of fungal growths and the nature of the growth recorded. Results: The recovery of filamentous fungi/black yeasts in 2013 (20.7%) was significantly less than 2015 (31.1%; difference 10.4%, 95% Cl: 3.4–17.4, P: 0.0037) and 2016 (28.8%; difference 8.1, 95% Cl: 1.2–15.0, P: 0.0281). The most commonly recovered fungi belonged to the genus Aspergillus and was cultured in 15–18% of patients across the 4 yrs. The increase in positive fungal cultures was attributed to a slight increase in the recovery of Aspergillus sp., but more so to significant increases in the culture of Exophiala sp. (P: 0.041) and Penicillium sp. (P: 0.0054). Conclusion: Over this 4yr period the epidemiology of fungal species detected in our CF patients appears relatively constant with the highest prevalence being Aspergillus sp. and Candida albicans with similar prevalence rates to other European cohorts. However, when differentiating potential pathogens from commensal organisms (Candida sp.) there was a significant increase in the number of filamentous fungi/black yeast covered. A limitation of this study is this data reflects only whether a patient has isolated a fungus at any time point during the year. Our ongoing projects are focussing on whether the frequency of fungi isolated from each individual is increasing and whether we can track the clinical effect of fungal colonisation. 144 Casting aspersions on Aspergillus – a comparison of clinical disease markers and therapy in patients with and without ABPA E. Spencer-Clegg1, R. Sapina-Vivo1, M.J. Walshaw1. 1Liverpool Heart & Chest Hospital, Adult CF Unit, Liverpool, United Kingdom Objectives: APBA is a complex disease which is difficult to diagnose and manage. We report on a review of clinical disease markers and management of ABPA. Methods: We compared age, FEV1 and IV ABX usage in 2016 for pts with a diagnosis of ABPA vs those without and those with a significantly raised Aspergillus specific IgG (>90 mg/mL) regardless of their diagnosis. In addition, we compared the Aspergillus specific IgG, FEV1 and IV ABX usage for ABPA patients on long term antifungals versus long term antifungals and steroids. Results: Of 303 pts reviewed, 65 (21%) had a diagnosis of ABPA at their last annual screen. A diagnosis of ABPA was associated with a higher Aspergillus specific IgG (92 vs 49mg/ml p = <0.05) and a lower FEV1 (72% vs 79% p = 0.05). There was no difference between age (29 vs 31 yrs p = NS) or number of IV ABX 27 vs 20days, p = NS). 52 pts had an Aspergillus specific IgG >90 mg/mL regardless of ABPA diagnosis and again this was associated with a lower FEV1 but no difference in IV ABX days or age (68% vs 79% p = 0.007, 25days vs 21days p = NS, 29 vs 31 yrs, p = NS). 27/52 pts had a diagnosis of ABPA and of these most were taking antifungals and/or oral steroids. Of the 65 pts with ABPA, 28 were taking long term antifungals (with pulsed steroid therapy as needed), vs 26 who were taking long term antifungals and steroids. Long term antifungals alone was associated with a lower Aspergillus specific IgG, fewer IV ABX and a higher FEV1 (75 vs 104mg/ml p = 0.02, 20 vs 39days p = 0.02, 81% vs 63% p = 0.001. There was no difference in pts’ age (26 vs 30 yrs p = NS). Conclusion: This study demonstrates the complexity of diagnosing and managing APBA since although FEV1 was lower for pts with ABPA and