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Raport științific - Etapa 2019

Phase II - Summary

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Confirmation of the carbapenemases production in the identified strains, determining the type of carbapenemase and establishing the phylogenetic relationships between the strains according to the isolation source

Reported period: 01.01.2019 - 31.12. 2019

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Phase II of the project aims to confirm the production of carbapenemases (CPs) in Pseudomonas aeruginosa and Acinetobacter baumannii strains isolated and identified from the aquatic environment through the wastewater and surface network, to determine the type of CP and to establish phylogenetic relationships between the strains according to the isolation source. For this purpose it was continued the collection of wastewater samples (seven wastewater treatment plants (WWTPs), as well as as from the hospital environment (untreated water containing hospital wastewater – influent/effluent of the collector channel from six hospitals from Romania, were determined the comparative levels of the microbial load in the collected samples from the different sampling points of the seven WWTPs (influent, effluent –wastewater (treated water before its discharge into the river), river water upstream and downstream from the WWTP, active sludge from the aeration tank and residual sludge) as well as from the collector channel of six hospital units, were identified the strains using automated and molecular methods, the resistance profiles, the type of CP/ESBL and the phylogenetic relationships between the strains were determined. During this phase, 46 samples were collected and analyzed, of which 14 samples of wastewater (influent, effluent), 10 samples of active and residual sludge from seven WWTPs, 14 samples of surface water upstream and downstream of the analyzed WWTPs, 8 wastewater sample from the collector  channel of six hospital units with representative territorial coverage that allowed the confirmation and identification of 76 non fermenting Gram negative rods (NFGNB) producing ESBL/CP from WWTP D; 45 strains from WWTP E, 47 from WWTP F; 59 from WWTP G; 25 from WWTP H; 24 strains from WWTP I and 33 strains from WWTP J; 108 NFGNB strains from the collector channel of the six hospital units as well as 26 strains of A. baumannii (n = 14) and P. aeruginosa (n = 12) isolated from intraspital infections. The analysis of antibiotic resistance profiles have shown a significant level of percentage of MDR and XDR both in the case of the nosocomial ones and those isolated from the wastewater and surface water network. The genotypic study of antibiotic resistance markers to carbapenems and ESBLs have demonstrated in most cases the presence of the same CPs and ESBLs circulating between the hospital environment and the analyzed aquatic environments (excluding unit S1 – were identified in P. aeruginosa blaVEB and in WWTP D - A. baumannii blaOXA-23, blaOXA-24, blaOXA-51, blaTEM, and blaPER). PFGE / ERIC-PCR techniques followed by MLST determined the association of the circulating strains between the hospital environment and the analyzed aquatic environments with epidemic clones associated with globally widespread metallo-β-lactamases (A. baumannii ST 2, ST 79 and P. aeruginosa ST 357) or previously reported in nosocomial strains in Romania (P. aeruginosa ST 233), the study of the horizontal transfer mechanisms of the CP between the hospital and the aquatic environment will be carried out in the next phase of the project.

 

Publication list

  • ISI papers

  1. Ibraheem S.A., Kadhem H.A., Hadeethi S.A., Jabir M.S., Grigore R., Popa M., Gheorghe I., Mihailescu D.F. Effects of silver nanoparticles on nosocomial Pseudomonas aeruginosa strains–an alternative approach for antimicrobial therapy. Rom Biotechnol Lett. 2019; 24(2): 286-293.

  2. Gheorghe I., Cristea V.C., Marutescu L., Popa M., Murariu C., Trusca B.S., Borcan E., Ghita M.C., Lazăr V., Chifiriuc M.C. Resistance and virulence features in carbapenem-resistant Acinetobacter baumannii community acquired and nosocomial isolates in Romania. Revista de Chimie, 2019, 10, 3502-3507.

  • Book chapter

  1. Gheorghe I., Cururțiu C., Dițu L.M. Therapeutic nanostructures: novel approaches. Book. Materials for Biomedical Engineering Bioactive Materials for Antimicrobial, Anticancer, and Gene Therapy. Elsevier. 2019. ISBN: 978-0-12-81, 442 p.

  • International Conferences

  1. Gheorghe I.; Czobor I; Avram I.; Alshaikhli N.; Pircalabioru-Gradisteanu G.; Negut A.C.; Marutescu L.; Lazar V.;  Chifiriuc M.C. Poster 0936. Genetic background of antibiotic resistance in multidrug-resistant Acinetobacter baumannii recovered from a wastewater treatment plant near Bucharest, Romania. 29th European Congress of Clinical Microbiology & Infectious Diseases, Amsterdam, Netherlands, from 13 - 16 April 2019.

  2. Gheorghe I.; Avram I.; Cristea V.C.; Borcan E.; Murariu C.; Czobor I.; Marutescu L.; Yu Feng Y.; Chifiriuc M.C.; Zhiyong Z. Poster 0943. Draft genome sequences of four clinical isolates of multidrug-resistant Acinetobacter baumannii from Romanian hospitals. 29th European Congress of Clinical Microbiology & Infectious Diseases, Amsterdam, Netherlands, from 13 - 16 April 2019.

  3. Czobor I., Avram I., Gheorghe I., Popa L.I., Neguț A.C., Marutescu L., Popa M., Pircalabioru-Gradisteanu G., Lazar V., Chifiriuc M.C. Poster 0460. Detection of the epidemic IncP-6 plasmid carrying blaKPC-2 in a multidrug-resistant Enterobcter kobei isolate from activated sludge of a wastewater treatment plant in Valcea County, Romania. 29th European Congress of Clinical Microbiology & Infectious Diseases, Amsterdam, Netherlands, from 13 - 16 April 2019.

  4. Czobor I., Popa L.I., Gheorghe I., Avram I., Neguț A.C., Pircalabioru-Gradisteanu G., Lazar V., Chifiriuc M.C. Molecular characterisation of multidrug-resistant Klebsiella pneumoniae strains isolated from wastewater treatment plant in Romania. Poster 1226. 29th European Congress of Clinical Microbiology & Infectious Diseases, Amsterdam, Netherlands, from 13 - 16 April 2019.

  5. Gheorghe I., Sarbu I., Borcan E., Murariu C., Cristea V.C., Marutescu  L., Czobor I., Curutiu C., Feng Y., Chifiriuc M.C., Zhiyong Z. Poster 201 - Draft genome sequences of four multidrug resistant Acinetobacter baumannii isolates from two Romanian hospitals. 8th Congress of European Microbiologists, FEMS 2019, Glasgow, Scotland, Glasgow, Scotland, 07-11.07.2019.

  6. Gheorghe I., Cristea V.C., Marutescu L., Popa M., Curutiu C., Murariu C., Trusca B.S., Borcan E., Ghita M.C., Lazar V., Chifiriuc M.C. Poster 206 Resistance and virulence features in carbapenem-resistant community- acquired and nosocomial Acinetobacter baumannii isolates in Romania. 8th Congress of European Microbiologists, FEMS 2019, Glasgow, Scotland, 07-11.07.2019.

  7. Gheorghe I., Sarbu I., Cristea V.C., Borcan E., Czobor I., Curutiu C., Feng Y., Murariu C., Lazar V., Chifiriuc M.C., Zong Zhiyong Z. Poster 202 Snapshot of resistance and clonality features in nosocomial and community acquired Acinetobacter baumannii strains. 8th Congress of European Microbiologists, FEMS 2019, Glasgow, Scotland, 07-11.07.2019.

Rezumatul etapei a II-a

Confirmarea producerii de carbapenemaze la tulpinile identificate, determinarea tipului de carbapenemază și stabilirea relațiilor filogenetice dintre tulpini în funcție de sursa de izolare

Perioada raportată: 01.01.2019 - 31.12. 2019

 

Etapa a II-a  proiectului îsi propune să confirme producerea de carbapenemaze (CP) la tulpini de Pseudomonas aeruginosa și Acinetobacter baumannii izolate și identificate din  rețeaua de apă uzată și de suprafață, să determine tipul de carbapenemază și să stabilească relațiile filogenetice dintre tulpini în funcție de sursa de izolare.  În acest scop a fost continuată activitatea de colectare de probe de apă uzată, s-au determinat nivelurile comparative ale încărcăturii microbiene în probele colectate din diferite puncte de prelevare ale celor șapte stații de epurare – SEAU (influent, efluent – apă uzată, amonte,  aval – apă de suprafață,  nămol activ din tancul de aerare și nămolul rezidual), precum și din canalul colector a șase unități spitalicești, au fost identificate tulpinile prin metode automate și moleculare, s-a determinat profilul de rezistență/sensibilitate la antibiotice, tipul de CP/β-lactamază de spectru extins (BLSE) și relațiile filogenetice dintre tulpini.  În cadrul acestei etape s-a realizat recoltarea și analiza a unui număr  de 46 de probe, dintre care 14 probe de apă uzată (influent, efluent), 10 probe de nămol activ si rezidual din șapte SEAU, 14 probe de apă de suprafață din amonte și aval de SEAU analizate, 8 probe de apă uzată din canalul colector a șase unități spitalicești cu acoperire teritorială reprezentativă. Au fost izolate și identificate 76 de tulpini de BGNNF producătoare de BLSE/CP din cadrul SEAU D; 45 tulpini din SEAU E, 47 de tulpini din SEAU F; 59 de tulpini din SEAU G; 25 de tulpini din SEAU H; 24 de tulpini din SEAU I și 33 de tulpini din SEAU J; 108 tulpini de BGNNF  producătoare de CP din canalele colectoare al celor șase unități spitalicești precum și 26 de tulpini de A. baumannii (n=14) și P. aeruginosa (n=12) izolate din infecții intraspitalicești. Analiza profilurilor de rezistență la antibiotice a evidențiat un procent semnificativ de tulpini multirezistente și cu rezistență extinsă atât în cazul celor nosocomiale, cât și al celor izolate din rețeaua de apă uzată și de suprafață. Studiul genotipic al markerilor de rezistență la carbapeneme și BLSE a demonstrat în majoritatea cazurilor prezența acelorași CP si BLSE circulante între mediul spitalicesc și în reteaua de apă uzată și de suprafață (exc. unitatea S1 – P. aeruginosa blaVEB iar în SEAU D – A. baumannii blaOXA-23, blaOXA-24, blaOXA-51, blaTEM, și blaPER). Tehnicile PFGE/ERIC-PCR urmate de tipizarea secvențelor multi-locus (MLST) au determinat apartenența tulpinilor circulante între mediul spitalicesc și rețeaua de apă uzată și de suprafață la clone epidemice asociate cu metalo-β-lactamaze larg răspândite la nivel global (A. baumannii ST 2, ST 79 și P. aeruginosa ST 357) sau anterior raportate la tulpini nosocomiale din România (P. aeruginosa ST 233), studiul mecanismelor de transfer pe orizontală a CP între mediul spitalicesc și cel avatic va fi realizat în etapa a III-a de derulare a proiectului.

Lista de publicații:

  • Articole ISI

  1. Ibraheem S.A., Kadhem H.A., Hadeethi S.A., Jabir M.S., Grigore R., Popa M., Gheorghe I., Mihailescu D.F. Effects of silver nanoparticles on nosocomial Pseudomonas aeruginosa strains–an alternative approach for antimicrobial therapy. Rom Biotechnol Lett. 2019; 24(2): 286-293.

  2. Gheorghe I., Cristea V.C., Marutescu L., Popa M., Murariu C., Trusca B.S., Borcan E., Ghita M.C., Lazăr V., Chifiriuc M.C. Resistance and virulence features in carbapenem-resistant Acinetobacter baumannii community acquired and nosocomial isolates in Romania. Revista de Chimie, 2019, 10, 3502-3507.

  • Capitole carte

  1. Gheorghe I., Cururțiu C., Dițu L.M. Therapeutic nanostructures: novel approaches. Book. Materials for Biomedical Engineering Bioactive Materials for Antimicrobial, Anticancer, and Gene Therapy. Elsevier. 2019. ISBN: 978-0-12-81, 442 p.

  • Participări conferințe internaționale

  1. Gheorghe I.; Czobor I; Avram I.; Alshaikhli N.; Pircalabioru-Gradisteanu G.; Negut A.C.; Marutescu L.; Lazar V.;  Chifiriuc M.C. Poster 0936. Genetic background of antibiotic resistance in multidrug-resistant Acinetobacter baumannii recovered from a wastewater treatment plant near Bucharest, Romania. 29th European Congress of Clinical Microbiology and Infectious Diseases, Amsterdam, Olanda, 13 - 16 Aprilie 2019. Autor principal conferinţă internaţională cu cotaţie ISI.

  2. Gheorghe I.; Avram I.; Cristea V.C.; Borcan E.; Murariu C.; Czobor I.; Marutescu L.; Yu Feng Y.; Chifiriuc M.C.; Zhiyong Z. Poster 0943. Draft genome sequences of four clinical isolates of multidrug-resistant Acinetobacter baumannii from Romanian hospitals. 29th European Congress of Clinical Microbiology & Infectious Diseases, Amsterdam, Olanda, 13 - 16 Aprilie 2019. Autor principal conferinţă internaţională cu cotaţie ISI.

  3. Czobor I., Avram I., Gheorghe I., Popa L.I., Neguț A.C., Marutescu L., Popa M., Pircalabioru-Gradisteanu G., Lazar V., Chifiriuc M.C. Poster 0460. Detection of the epidemic IncP-6 plasmid carrying blaKPC-2 in a multidrug-resistant Enterobcter kobei isolate from activated sludge of a wastewater treatment plant in Valcea County, Romania. 29th European Congress of Clinical Microbiology & Infectious Diseases, Amsterdam, Netherlands, from 13 - 16 April 2019. Co-autor conferinţă internaţională cu cotaţie ISI.

  4. Czobor I., Popa L.I., Gheorghe I., Avram I., Neguț A.C., Pircalabioru-Gradisteanu G., Lazar V., Chifiriuc M.C. Molecular characterisation of multidrug-resistant Klebsiella pneumoniae strains isolated from wastewater treatment plant in Romania. Poster 1226. 29th European Congress of Clinical Microbiology & Infectious Diseases, Amsterdam, Netherlands, from 13 - 16 April 2019. Co-autor conferinţă internaţională cu cotaţie ISI.

  5. Gheorghe I., Sarbu I., Borcan E., Murariu C., Cristea V.C., Marutescu  L., Czobor I., Curutiu C., Feng Y., Chifiriuc M.C., Zhiyong Z. Poster 201 - Draft genome sequences of four multidrug resistant Acinetobacter baumannii isolates from two Romanian hospitals. 8th Congress of European Microbiologists, FEMS 2019, Glasgow, Scoția, 07-11.07.2019. Autor principal.

  6. Gheorghe I., Cristea V.C., Marutescu L., Popa M., Curutiu C., Murariu C., Trusca B.S., Borcan E., Ghita M.C., Lazar V., Chifiriuc M.C. Poster 206 Resistance and virulence features in carbapenem-resistant community- acquired and nosocomial Acinetobacter baumannii isolates in Romania. 8th Congress of European Microbiologists, FEMS 2019, Glasgow, Scoția, 07-11.07.2019. Autor principal

  7. Gheorghe I., Sarbu I., Cristea V.C., Borcan E., Czobor I., Curutiu C., Feng Y., Murariu C., Lazar V., Chifiriuc M.C., Zong Zhiyong Z. Poster 202 Snapshot of resistance and clonality features in nosocomial and community acquired Acinetobacter baumannii strains. 8th Congress of European Microbiologists, FEMS 2019, Glasgow, Scoția, 07-11.07.2019. Autor principal.

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