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Results: The percentage share of NFGNB other than PA was 28%. The most common isolated NFGNB were: Stenotrophomonas 10%, Achromobacter 5%, non-aeruginosa Pseudomonas 5%, Sphingomonas 3%, Acinetobacter 2% and others 3%. NFGNB showed variable sensitivity to carbapenems (42–89%), fluoroquinolones (13–100%), aminoglycosides (22–100%), cephalosporins (23–78%), colistin (84–100%), co-trimoxazole (75–97%). Conclusion: We observed significant increase of NFGNB other than PA in colonization of the airways of CF pts. Prolonged incubation and using a selective media increases the frequency of NFGNB isolation. Mass spectrometry is a reliable alternative to conventional biochemical tests used in identification of rarely isolated NFGNB. For some NFGNB species it is required to determine the MIC values to interpret their susceptibility.

throat in younger children. The results were evaluated for two years period. Results: 42.6% of patients had positive nasal exudates, the most frequent microbe being Staphylococcus aureus (22.2%, unfortunately 14.8% had MRSA). It was followed by Streptococcus pneumoniae, found in 18.5% of patients, and Pseudomonas in 11% of patients. In patients with exacerbation, we evaluated the correlation of nasal microbiota with sputum or swab samples, revealing a small correlation index (only in 14.8% OR = 0.55, 95% CI: 0.2013 to 1.5213) between the nasal microbes and exacerbation’s ethiology. Conclusion: An important percentage of nasal microbes was found in our patients, but with a low correlation with the etiologic agent of pulmonary exacerbations. Nevertheless, a higher importance should be given for the effect of nasal microbiota in the future.

61 Composition of the upper aero-digestive and lower respiratory tracts bacterial microbiota in CF patients

63 Respiratory infections in children with cystic fibrosis (CF). Relationship with clinical and functional features

R. Rivas Caldas1 , J. Mounier2 , G. Michel3 , G. Rault4 , S. Boisrame1 , G. Barbier1 . 1 LUBEM B-V, Universit´e de Bretagne Occidentale (UBO), Brest, France; 2 LUBEM, ESIAB, Plouzane, France; 3 UMR-7139 CNRS Roscoff, Roscoff, France; 4 CRCM de Roscoff, Centre de Perharidy, Roscoff, France

C. Burattini1 , C. Tripodi1 , C. Spaggiari1 , F. Longo1 , I. Cortina1 , G. Pisi1 . 1 Cystic Fibrosis Unit Parma University Hospital, Parma, Italy

Objectives: There is lack of knowledge about the upper aero-digestive tract (UADT) and lower respiratory tract (LRT) microbiota in CF patients. Changes in the microbiota composition in CF patients could influence the pathophysiology of the respiratory tract disease. Moreover, It has been reported that the UADT is a secondary reservoir for respiratory pathogens. The aim is to analyze the relationship and possible dynamic variations between the UADT and LRT microbiota using the NGS. Methods: Samples were obtained at two different times from six CF children and two adults. Three groups were defined according to P. aeruginosa lungs colonization status: not-colonized young, notcolonized “old” and colonized patients. The V3 and V5 hypervariable region of 16SrRNA gene were amplified. Data set was processed and analyzed by different pipelines. Results: Bacterial diversity was high. Similar bacteria genera were found in the UADT and the LRT samples. The core microbiota was different between patients groups. The reliability of the results is in progress by comparing results obtained by the different pipelines. Ecotypes depending on P. aeruginosa pulmonary colonization and patients’ age will be presented. Conclusion: UADT and LRT microbiota have similar genera diversity. Further studies are necessary to determine if P. aeruginosa colonization would be the reason of this microbiota modification, or the contrary, the microbiota modification would be the cause of P. aeruginosa colonization. Nevertheless, in our knowledge, this is the first study that analyzes and compares CF UADT and LRT samples by using NGS. 62 Evaluation of nasal microbiota in cystic fibrosis children I.M. Ciuca1,2 , L.L. Pop1,2 , F. Horhat3 , G. Iovanescu4 , C. Oancea5 , Z. Popa2 . 1 University of Medicine and Pharmacy Victor Babes, Pediatric Department, Timisoara, Romania; 2 National Cystic Fibrosis Centre, Timisoara, Romania; 3 University of Medicine and Pharmacy ‘Victor Babes’, Microbiology Department, Timisoara, Romania; 4 University of Medicine and Pharmacy Victor Babes, Otorhinolaryngology Department, Timisoara, Romania; 5 University of Medicine and Pharmacy ‘Victor Babes’, Pulmonology Department, Timisoara, Romania Objectives: Considering the risk of pulmonary infections in children with CF in the context of carriage status for varied microbes, we considered important for our patients to evaluate the nasal microbiota status among our patients. Methods: A cross-sectional study included fifty-four CF patients monitored in our center were evaluated every six months; at every visit, the clinical status, biological assessment and other investigations were performed according to guidelines. Also, every 3 months, the bacteriological investigations for nasal secretion were collected, besides sputum culture (required mostly in children older than 7 years) or swab

Objectives: The aim of the study was to assess the respiratory infections in a paediatric CF population and their correlations with clinical and functional data. Methods: We retrospectively studied 37 patients (22 males), age range 6 months to 16 years, recruited between 1998 and 2014 in our CF Clinic at Parma University Hospital. We recorded the following parameters: sex, age, respiratory infections, BMI, FEV1 , FVC, FEV1 /FVC, SaO2 , number of pulmonary exacerbations and IV antibiotic courses. For the statistical analysis, unpaired Student T test, contingency table and c2 analysis, and linear regression were used. A p value <0.05 was considered as significant. Results: The most common pathogens were S. aureus (52%), P. aeruginosa (19%), E. coli (10%), and C. albicans (18%) and A. fumigatus (11%), respectively. In comparison with infected patients, the germ-free patients were significantly younger (3.2±2.0 yr vs 6.2±3.8 yr, p < 0.001) and had lower number of pulmonary exacerbations (0.06±0.2 vs 0.2±0.4, p < 001) and IV antibiotic courses (0.1±0.4 vs 0.3±0.5, p < 0.001). Children infected by P. aeruginosa and A. fumigatus showed worse lung function than non-infected ones. Conclusion: Our study shows that many pathogens may be involved in the respiratory infections in a paediatric CF population. Respiratory infections were associated to age and to the pulmonary exacerbations rate, and, when due to P. aeruginosa and A. fumigatus, with a worse lung function. Prevention of infections and early and specific antimicrobial treatments are the cornerstone in CF care of pulmonary disease. 64 Chronic pulmonary infection with Pseudomonas aeruginosa in relation to F508del mutation in patients with cystic fibrosis S. Sciuca1 , L. Balanetchi1 , C. Grigore1 , O. Dimitrova1 , A. Cotoman2 , A. Ceban2 . 1 State Medical and Pharmaceutical University, Chisinau, Moldova, Republic of ; 2 Mother and Child Institute, Chisinau, Moldova, Republic of Objectives: Evaluation of the frequency of chronic pulmonary infection with P. aeruginosa in cystic fibrosis (CF) patients with genotype F508del. Methods: The study included 69 CF patients who were examined at P. aeruginosa pulmonary infection. The diagnosis of CF was confirmed by a sweat test (Macroduct, USA), mutation of CFTR. In 45 patients (65.2%) has been identified F508del mutation, of which 27 cases (39.1%) – homozygous (F508del/F508del) and 18 cases (26.1%) – heterozygous (F508del/non-F508del), while 24 patients (34.8%) were the non-F508del group. Results: P. aeruginosa pulmonary infection was confirmed in 76.2% of CF patients with F508del mutation, including 40.5% with genotype F508del/F508del. In CF patients CFTR non-F508del pulmonary infection with P. aeruginosa was detected in only 23.8%. In the group of CF patients without P. aeruginosa infection predominates CFTR genotype non-F508del (51.9%) and patients with genotype F508del/F508del constituted 37.0%, heterozygous patients F508del – 11.1%. The analysis

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demonstrated that the F508del mutation is responsible for chronic lung infections with P. aeruginosa in CF patients. Conclusion: Patients with cystic fibrosis genotype CFTR F508del performed more frequently P. aeruginosa chronic pulmonary infection than non-F508del CF patients. 65 Relationship between age of cystic fibrosis diagnosis and microbiological tests results in Lithuanian children CF centre V. Radziuniene1 , S. Dumcius1 , O. Kinciniene2 , J. Svarauskaite2 . 1 Vilnius City Clinical Hospital, Vilnius, Lithuania; 2 Vilnius University, Faculty of Medicine, Vilnius, Lithuania Objectives: Children with cystic fibrosis predisposed to grow a variety of bacterial pathogens. This study aimed to determine microbiological diversity and it correlation with the age of cystic fibrosis diagnosis in CF patients. The study was performed in children CF centre in Vilnius City Clinical Hospital. Methods: We analysed case histories of 30 patients under 18 years old during 1 year period in January–December 2015. All patients were divided into 3 groups according to their age of diagnosis (under 1 year; 1–5 years; over 5 years). Patient’s microbiological diversity in clinical materials (sputum, cough swabs, BAL) was analysed. Results: 30 patients had at least one positive sample over the study period. Out of 97 bacterial cultures grown there were 81 positive samples with 31 different species. Pseudomonas aeruginosa was found in 50.62% of samples. S. aureus was found at least once in 51.85% of samples, only one patient had MRSA. Stenotrophomonas maltophilia was isolated in 9.88%, Achromobacter – 3.3%, Candida – 14.81% of samples. Younger age of diagnosis is associated with higher probability of Pseudomonas aeruginosa growth but not S. aureus or other pathogens. Conclusion: Younger age of diagnosis lead to higher probability of Pseudomonas aeruginosa growth. The most common isolated pathogens were S. aureus and Pseudomonas aeruginosa. 66 Microbiological monitoring of lower respiratory tract infection in patients with cystic fibrosis M.Y. Chernukha1 , I.A. Shaginyan1 , L.R. Avetisyan1 , E.A. Siyanova1 , G.V. Alekseeva1 , O.S. Medvedeva1 , N. Kashirskaya2 , E.I. Kondratieva2 , N. Kapranov2 . 1 N.F. Gamaleya Federal Research Centre of Epidemiology and Microbiology, Moscow, Russian Federation; 2 Research Centre for Medical Genetics, Moscow, Russian Federation Background: Infection in lungs is the main cause of lethal outcomes in 90–95% of cystic fibrosis (CF) patients. The predominant pathogens are Staphylococcus aureus, Pseudomonas aeruginosa, Burkholderia cenocepacia and Achromobacter xylosoxidans. Objectives: Monitoring of S. aureus, P. aeruginosa, B. cenocepacia and A. xylosoxidans lung infections in CF patients in Russia. Methods: Diagnostic algorithm included phenotypic and molecular techniques – antimicrobial susceptibility testing, RAPD-PCR, MLST. Results: The effectiveness of antibiotic therapy of S. aureus infection was 74%, P. aeruginosa – 58.8%, B. cenocepacia – 0%, A. xylosoxidans – 0%. 5 year monitoring showed that the genotype of S. aureus changed in 4% of cases. Due to antibiotic therapy the initial strain was eliminated, then recolonization with a new strain occurred. During P. aeruginosa infection the genotype changed in 19.1% of patients. During B. cenocepacia and A. xylosoxidans infection the genotype did not change. The strains isolated earlier were genetically identical to the strains isolated after 5 years. Analysis by MALDI-TOF-MS of early and late strains of identical genotype showed protein variability. Epidemic clones of S. aureus, P. aeruginosa, B. cenocepacia and A. xylosoxidans were isolated. The contamination by B. cenocepacia ST709 (identified in 67.6% CF patients) and ST208 (identified in 12.4%) is supposed to occur during hospitalizations. Conclusion: Antimicrobial therapy is effective in treatment of S. aureus infection, less effective in P. aeruginosa infection and is almost ineffective in B. cenocepacia and A. xylosoxidans infections. These infections can be associated with health care.

67 Pilot study on bacterial contamination of inhalation devices in cystic fibrosis 2 E. Coirier Duet1 , B. Delaisi2 , C. Doit2 , M. Gerardin ´ , V. Houdoin2 , S. Gonsseaume2 , D. Grenet3 , S. de Miranda3 , E. Farfour3 , A. Munck3 . 1 CF Center for Children-Versailles, 78, Le Chesnay, France; 2 CF Center for Children, Robert Debr´e Paris 3, Paris, France; 3 CF Center for Adults, Suresnes, France

Aim: To evaluate the bacterial contamination of inhalation devices in current conditions of use, in cystic fibrosis. Methods: This study was conducted in 2 CF centers. Patients receiving an inhaled treatment were contacted by phone the day before the consultation to bring the dispositif(ves) in a clean linen, without modifications of routine maintenance. During consultation, devices were controlled by swabbing with wet swab, by the CF staff. Swabs were inoculated on a non-selective medium (chocolate agar). Simultaneously a sputum sample was collected (or a pharangeal sample) for microbiology analysis. Results: 46 patients (27 children). Chronic colonisation with Pseudomonas aeruginosa was present in 5% of children and 80% of adults; Methicillin-resistant Staphyloccus aureus concerned only 10% of adults. Devices were: 12 inhalation chambers, 19 reusable nebulizers, 34 powder dispositives or metered-dose nebulizer (18 for adults). There was a wide variety of modalities for deterging and drying the devices on routine. Scrubbings were sterile for 75% of the devices (70% for children; 93% for adults). When non sterile, the contamination was from the oropharyngeal or cutaneous flora. Discussion: We had very little positive cultures. Our bacterial results were identical when we compared wet swabs and culture medium obtened by rinsing devices and the exploitation of the devices was quite shortly after the last use. Conclusions: This pilot study found a very low rate of contamination of devices. In case of contamination, there was no relation with the bacteria from the sputum and no airways pathogens for CF patients. We aim to extend our analysis to other CF centers. 68 Frequency and persistence of CF airway pathogens from diagnosis to age 16 years K. De Boeck1 , F. Vermeulen1 , M. Boon1 , T. Havermans1 , M. Proesmans1 . 1 University of Leuven, Pediatric Pulmonology, Leuven, Belgium Little is known about the relative frequency and persistence of S. maltophilia (Sm), A. xylosoxidans (Ax) and B. cepacia (Bc) in the CF airway compared to S. aureus (Sa) and P. aeruginosa (Pa). We thus explored this during childhood. Data were extracted from the hospital electronic records. Only patient years (PY) with ≥4 airway cultures in different months were explored. Sa, Pa, Sm, Ax and Bc were evaluated in each PY as present/absent and considered chronic if present in ≥50% of cultures. Age at first occurrence of pathogen was noted; clearance was defined as not isolated in ≥3 consecutive years. In 43 subjects born 1994 to 1999 and followed until age 16, 81% of all PY could be analysed. Chronic infection was detected in 291/555 (52%) PY: with Sa 250 (45%), with Pa 46 (8%), with Ax 18 (3%), with Sm 10 (2%); 27 (5%) PY had chronic infection with 2 pathogens (Sa+Pa in 17/27). When ‘present in the year’ was considered, Sa was grown in 67% of PY, Pa in 25%, Ax in 4%, Sm in 5% and Bc in 1%; in 23% of PY none of these pathogens were grown. First isolation of Sm, Ax or Bc occurred in 26/43 subjects: 3 events in 2–6 yr olds, 13 in 7–11 yr, 24 in 12–16 yr olds. Sm was most frequent in 7–11 yr (8/15), Ax (10/16) and Bc (4/4) in 12–16 yr old. Clearing of pathogen ± treatment decreased from 3/3 in the youngest, over 8/13 to 9/24 in the 12–16 yr old. %FEV1 at age 16 yr was lower in pts who ever had chronic Pa/Ax/Sm infection PY (76±5 vs 89±3; p = 0.02) and correlated (r = −0.43; p = 0.004) with the number of chronic PY. In conclusion, during childhood Sa is the most frequently isolated pathogen. The risk of persistent infection with Sm, Ax and Bc increases towards adolescence.