{"id":94,"date":"2018-09-01T14:05:59","date_gmt":"2018-09-01T14:05:59","guid":{"rendered":"https:\/\/bios.unibuc.ro\/?p=94"},"modified":"2023-12-08T13:08:45","modified_gmt":"2023-12-08T11:08:45","slug":"pn-iii-p3-3-1-pm-ro-cn-2018-0147","status":"publish","type":"post","link":"https:\/\/bios.unibuc.ro\/?p=94","title":{"rendered":"PN-III-P3-3.1-PM-RO-CN-2018-0147"},"content":{"rendered":"\n<p class=\"wp-block-paragraph\"><strong>Studii in silico \u0219i experimentale ale unor platforme genetice mobile de rezisten\u021b\u0103 multipl\u0103 la antibiotice<\/strong>.<\/p>\n\n\n\n<div style=\"height:10px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<hr class=\"wp-block-separator has-alpha-channel-opacity\"\/>\n\n\n\n<div class=\"wp-block-buttons is-layout-flex wp-block-buttons-is-layout-flex\">\n<div class=\"wp-block-button has-custom-width wp-block-button__width-100\"><a class=\"wp-block-button__link wp-element-button\" href=\"https:\/\/bios.unibuc.ro\/wp-content\/uploads\/2023\/12\/ro_cn_2019.pdf\" target=\"_blank\" rel=\"noreferrer noopener\">Raport stiintific Etapa I- 2018<\/a><\/div>\n\n\n\n<div class=\"wp-block-button has-custom-width wp-block-button__width-100\"><a class=\"wp-block-button__link wp-element-button\" href=\"https:\/\/bios.unibuc.ro\/wp-content\/uploads\/2023\/12\/rst_etapa_ii.pdf\" target=\"_blank\" rel=\"noreferrer noopener\">Raport de activitate<\/a><\/div>\n\n\n\n<div class=\"wp-block-button has-custom-width wp-block-button__width-100\"><a class=\"wp-block-button__link wp-element-button\" href=\"https:\/\/bios.unibuc.ro\/wp-content\/uploads\/2023\/12\/ro_cn_2019.pdf\" target=\"_blank\" rel=\"noreferrer noopener\">Raport final de activitate<\/a><\/div>\n<\/div>\n\n\n\n<hr class=\"wp-block-separator has-alpha-channel-opacity\"\/>\n\n\n\n<h2 class=\"wp-block-heading has-vivid-green-cyan-background-color has-background has-medium-font-size\"><em>(RO)&nbsp;RAPORT FINAL DE ACTIVITATE<\/em><\/h2>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Titlul proiectului&nbsp;<\/strong><em>STUDII IN SILICO \u0218I EXPERIMENTALE ALE UNOR PLATFORME GENETICE MOBILE DE REZISTEN\u021a\u0102 MULTIPL\u0102 LA ANTIBIOTICE&nbsp;<\/em>[<em>RESCUE -RESISTANCE]<\/em><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>contractului &#8211;&nbsp;<\/strong>PN-III-P3-3.1-PM-RO-CN-2018-0147 (CONTRACT NR. 5 BM)<\/li>\n\n\n\n<li><strong>Anul finaliz\u0103rii &#8211;&nbsp;<\/strong>2019<\/li>\n\n\n\n<li><strong>Durata proiectului&nbsp;<\/strong>18 luni<\/li>\n\n\n\n<li><strong>Partener rom\u00e2n<sup>&nbsp;&#8211;&nbsp;<\/sup><\/strong>Universitatea din Bucure\u0219ti, Reprezentant legal Rector prof.dr. Mircea Dumitru<\/li>\n\n\n\n<li><strong>Director de proiect &#8211;&nbsp;<\/strong>univ. dr. Mariana Carmen Chifiriuc<strong>&nbsp;<\/strong><\/li>\n\n\n\n<li><strong>Partener str\u0103in &#8211;&nbsp;<\/strong>West China Hospital, Sichuan University, Director de proiect prof.dr. Xiaohui Wang<\/li>\n\n\n\n<li><strong>Obiective generale urm\u0103rite<\/strong><\/li>\n<\/ul>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; Obiectivul general al proiectului este studiul evolu\u021biei locale a rezisten\u021bei la antibiotice (AR) la izolate clinice \u0219i din re\u021beaua de ap\u0103 uzat\u0103 de bacili Gram-negativi, multirezistente la antibiotice (MDR), prin studiul suportului genetic al AR \u0219i al transferabilit\u0103\u021bii genelor de rezisten\u021b\u0103 la antibiotice (ARG) prin intermediul elementelor genice mobile (MGE), pentru o mai bun\u0103 cunoa\u0219tere a epidemiologiei moleculare a AR \u00een Rom\u00e2nia (diferen\u021bierea clonelor AR emergente local de cele regionale \/ globale) \u0219i \u00eembun\u0103t\u0103\u021birea&nbsp; managementului local al AR.<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Descrierea \u015ftiin\u0163ific\u0103 \u015fi tehnic\u0103 a rezultatelor \u015fi gradul de realizare a obiectivelor<\/strong><\/li>\n<\/ul>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Toate obiectivele \u0219i activit\u0103\u021bile propuse au fost \u00eendeplinite<\/strong>.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; Astfel, \u00een cadrul acestui proiect, au fost analizate 101 de tulpini bacteriene Gram-negative cu fenotip MDR (respectiv 83 tulpini de&nbsp;<em>Klebsiella pneumoniae<\/em>, 12 tulpini de&nbsp;<em>Acinetobacter baumannii<\/em>, 3 tulpini de&nbsp;<em>Escherichia coli,&nbsp;<\/em>&nbsp;1 tulpin\u0103 de&nbsp;<em>Serratia<\/em>&nbsp;sp., 1 tulpin\u0103 de&nbsp;<em>Enterobacter kobei<\/em>&nbsp;\u0219i 1 tulpin\u0103 de&nbsp;<em>Providencia<\/em>&nbsp;sp.). A fost investigat\u0103 prin experimente de PCR simplex \u0219i multiplex prezen\u021ba genelor de rezisten\u021b\u0103 la carbapeneme, dar \u0219i a genelor de AR ce confer\u0103 fenotip BLSE. Experimentele PCR la tulpinile de&nbsp;<em>Enterobacteriaceae&nbsp;<\/em>au eviden\u021biat prezen\u021ba \u00een ordine descresc\u0103toare a genelor&nbsp;<em>bla<\/em><sub>SHV,&nbsp;<\/sub><em>bla<\/em><sub>NDM-1<\/sub>,&nbsp;<em>bla<\/em><sub>OXA-48<\/sub>,&nbsp;<em>bla<\/em><sub>TEM&nbsp;<\/sub>si&nbsp;<em>bla<\/em><sub>KPC<\/sub>. \u00cen ceea ce prive\u0219te tulpinile de&nbsp;<em>A. baumannii<\/em>&nbsp;experimentele PCR au evidentiat prezenta genei&nbsp;<em>bla<\/em><sub>oxa-51&nbsp;<\/sub>la toate tulpinile analizate, urmat\u0103 de&nbsp;<em>bla<\/em><sub>OXA-23,<\/sub><em>bla<\/em><sub>SHV,&nbsp;<\/sub><em>bla<\/em><sub>VEB<\/sub>,&nbsp;<em>bla<\/em><sub>TEM,&nbsp;<\/sub><em>bla<\/em><sub>NDM-1&nbsp;<\/sub>si&nbsp;<em>bla<\/em><sub>VIM<\/sub>.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; Investigarea transferabilit\u0103\u021bii AR s-a realizat prin experimente de conjugare realizate at\u00e2t \u00een mediu lichid (Liquid mating), c\u00e2t \u0219i pe mediu solid (Filter mating). Selec\u021bia transconjugan\u021bilor s-a realizat pe mediu&nbsp; MacConkey suplimentat cu azida de sodiu (150\u00b5g\/ml) \u0219i imipenem (1 \u00b5g\/ml), cat si pe mediu BHI (Brain heart infusion) suplimentat cu azid\u0103 de sodiu (150\u00b5g\/ml) \u0219i imipenem (1 \u00b5g\/ml). \u00cen cazul tulpinilor de&nbsp;<em>A. baumannii<\/em>, selec\u021bia s-a efectuat pe mediu BHI suplimentat cu rifampicin\u0103 (250mg\/l) \u0219i meropenem (1 \u00b5g\/ml) ca agen\u021bi de selec\u021bie.&nbsp; Majoritatea tulpinilor de&nbsp;<em>Enterobacteriaceae<\/em>&nbsp;\u0219i trei dintre tulpinile de&nbsp;<em>A. baumannii<\/em>&nbsp;incluse \u00een experimentele de conjugare s-au dovedit a fi tulpini conjugative, cea mai mare parte a lor conjug\u00e2nd cu precadere pe mediu solid (filtru). Rezultatele PCR ob\u021binute pentru transconjugan\u021bi au eviden\u021biat transferul unora dintre genele care confer\u0103 rezisten\u021ba la antibioticele \u03b2-lactamice, \u00een cazul uneia dintre tulpinile de&nbsp;<em>K. pnumoniae<\/em>&nbsp;fiind eviden\u021biate trei gene transferate simultan, ceea ce sugereaz\u0103 localizarea acelor gene \u00een acelea\u0219i elemente genetice mobile \u0219i posibilitatea disemin\u0103rii acestora pe orizontal\u0103.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; \u00cen a doua etap\u0103 a proiectului s-a realizat secven\u021bierea unor gene de rezisten\u021b\u0103 sau genomuri bacteriene de&nbsp;<em>K. pneumoniae<\/em>,&nbsp;<em>A. baumannii<\/em>,&nbsp;<em>E. coli<\/em>&nbsp;\u0219i&nbsp;<em>E. kobei&nbsp;<\/em>\u0219i analiza bioinformatic\u0103, at\u00e2t a datelor de secven\u021biere ob\u021binute, c\u00e2t \u0219i a conforma\u021biei \u0219i interac\u021biunii proteinelor codificatoare pentru ARGs. S-a realizat secven\u021bierea&nbsp;<em>Next Generation Sequencing &#8211;&nbsp; pair-end whole genome sequencing \u2013 MiSeq System Sequencer, Illumina<\/em>, iar analiza bioinformatic\u0103 a produ\u0219ilor de secven\u021biere (cu ajutorul&nbsp;<em>software<\/em>-urilor de asamblare \u0219i adnotare SPAdes, CARD, BLAST, ISFinder, PlasmidFinder, ResFinder, VirulenceFinder, INTEGRALL) \u00een ceea ce priveste prezen\u021ba ARGs&nbsp; a demonstrat faptul c\u0103 tulpinile luate \u00een studiu au prezentat gene de rezisten\u021b\u0103 pentru un num\u0103r ridicat de clase de antibiotice (\u00eentre 4 \u0219i 8 clase). \u00cen ceea ce prive\u0219te prezen\u021ba plasmidelor asociate cu AR pentru tulpinile de&nbsp;<em>Enterobacteriaceae<\/em>&nbsp;selectate, a fost eviden\u021biat\u0103 prezen\u021ba a opt tipuri de plasmide, respectiv ColE10, IncA\/C2, IncFIB(K),&nbsp; IncFII(Yp), IncHI1A(CIT), IncHI1B(CIT), IncP6, IncX1 la o tulpin\u0103 de&nbsp;<em>E. coli<\/em>, cinci Col440II, ColRNAI, IncFII(K), IncFII(Yp), IncR \u0219i respectiv trei tipuri, ColpVC, IncFII(Yp), IncL\/M (pOXA-48) de plasmide &nbsp;la tulpini de&nbsp;<em>K. pneumoniae<\/em>. Cele 3 tulpini de&nbsp;<em>Enterobacteriaceae<\/em>&nbsp;secven\u021biate prin NGS au apar\u021binut clonelor ST101 si respectiv ST383 (tulpinile de&nbsp;<em>K. pneumoniae<\/em>) \u0219i ST 378 (tulpina de&nbsp;<em>E. coli<\/em>). &nbsp;<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; Studiile conforma\u021bionale au indicat conforma\u021bii ale proteinelor OXA-23 \u0219i SHV-2 codificate de alelele identificate \u00een urma analizei datelor de secven\u00e2iere la tulpinile circulante, identice cu cele existente \u00een bazele de date interna\u021bionale.<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Impactul proiectului<\/strong><\/li>\n<\/ul>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; &nbsp; Colaborarea cu China (vizite \u0219tiin\u021bifice, organizarea de ateliere interna\u021bionale) a contribuit esen\u021bial la dezvoltarea resursei umane, oferind oportunitatea specializ\u0103rii tinerilor membri ai echipei din Rom\u00e2nia \u00een tehnica&nbsp;<em>Next Generation Sequencing<\/em>&nbsp;\u2013MiSeq System Sequencer, Illumina \u0219i \u00een analiza bioinformatic\u0103 a datelor de secven\u021biere; aceast\u0103 expertiz\u0103 va putea fi aplicat\u0103 imediat \u00een alte dou\u0103 proiecte de cercetare interna\u021bionale \u0219i unul na\u021bional \u00een curs de desf\u0103\u0219urare, contribuing esen\u021bial la &nbsp;ob\u021binerea unor rezultate valoroase, prin utilizarea unei metodologii de ultim\u0103 or\u0103.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; Cunoa\u0219terea epidemiologiei moleculare a AR tulpinilor de BGN circulante \u00een Rom\u00e2nia at\u00e2t \u00een sectorul clinic, c\u00e2t \u0219i \u00een mediul acvatic (diferen\u021bierea clonelor AR emergente local de cele regionale \/ globale) va contribui la \u00eembun\u0103t\u0103\u021birea&nbsp; managementului local al AR.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; Rezultatele ob\u021binute \u00een cadrul acestui proiect sunt promi\u021b\u0103toare \u0219i vor face obiectul a cel pu\u021bin dou\u0103 publica\u021bii majore (una dintre acestea axat\u0103 pe tulpinile MDR de&nbsp;<em>Klebsiella<\/em>&nbsp;sp., ia a doua pe cele de&nbsp;<em>A. baumannii<\/em>) \u0219i a trei rezumate care vor fi trimise la 30th European Congress of Clinical Microbiology &amp; Infectious Diseases (ECCMID), Paris, Fran\u021ba, 18-20 Aprilie 2020.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>INDICATORI FINALI AI PROIECTULUI<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Vizite de lucru efectuate pe durata proiectului bilateral<\/strong><\/li>\n\n\n\n<li><em>Vizita echipei chineze<\/em>&nbsp;(director proiect: Wang Xiaohui, membrii echimei: Zong Zhiyong, Feng Yu, Liu Lina) \u00een Rom\u00e2nia: 31.05-11.06.2019<\/li>\n<\/ul>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; \u00cen cadrul acestei vizite a echipei chineze la Institutul de Cercet\u0103ri al Universit\u0103\u021bii din Bucure\u0219ti s-au stabilit protocoalele de urmat \u00een cadrul proiectului \u0219i de asemenea au fost selectate tulpinile de analizat. De asemenea, s-a realizat instruirea echipei rom\u00e2ne de c\u0103tre partenerii chinezi cu privire la realizarea experimentelor de conjugare bacterian\u0103 \u0219i selectare a transconjugan\u021bilor, dar \u0219i \u00een ceea ce prive\u0219te analiza bioinformatic\u0103 a datelor, respectiv asamblarea \u0219i adnotarea primar\u0103 \u0219i detaliat\u0103 a genomurilor bacteriene \u0219i compararea acestora.<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><em>Vizita echipei rom\u00e2ne<\/em>&nbsp;(membrii echipei: A.C. Ilda Barbu, CS Marius Surleac, drd. Laura Ioana Popa, lect dr. Irina Gheorghe) \u00een China &#8211; Chengdu: 15-28.09.2019<\/li>\n<\/ul>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; \u00cen cadrul acestei vizite au fost realizate experimente de conjugare bacterian\u0103, selectare a transconjugan\u021bilor \u0219i analiz\u0103 a acestora, secven\u021bierea unor genomuri bacteriene de interes \u0219i analiza bioinformatic\u0103 a datelor.<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Stagii de specializare<\/strong><\/li>\n<\/ul>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; Echipa rom\u00e2n\u0103 (.C. Ilda Barbu, CS Marius Surleac, drd. Laura Ioana Popa, lect dr. Irina Gheorghe) a participat la un workshop organizat de institu\u021bia gazd\u0103 (Universitatea Sichuan, prof. dr. Zong Zhiyong) referitor la bioinformatica asambl\u0103rii \u0219i adnot\u0103rii de genomuri bacteriene.<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Diseminarea rezultatelor<\/strong><\/li>\n<\/ul>\n\n\n\n<p class=\"wp-block-paragraph\"><em>Articole ISI<\/em><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Mitache M.M., Banu O., Borcan E., Gheorghe I., Marutescu M., Popa M., Predescu M., Rusu E., Nemes R., Chifiriuc M.C. 2018.&nbsp;<a href=\"http:\/\/scholar.google.com\/scholar?cluster=18418457897074986975&amp;hl=en&amp;oi=scholarr\">Resistance and Virulence Patterns in Gram Negative and Gram Positives Rods Isolated from the Hospital Environment in Bucharest, Romania<\/a>&nbsp;Revista De Chimie 69 (11), 4026-4032<\/li>\n<\/ul>\n\n\n\n<p class=\"wp-block-paragraph\"><em>Articole (BDI)<\/em><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Podgoreanu P., Negrea S., Buia R., Delcaru C., Trusca S.B., Lazar V., Chifiriuc M.C. 2019.&nbsp;<a href=\"http:\/\/scholar.google.com\/scholar?cluster=16293756831297448303&amp;hl=en&amp;oi=scholarr\">Alternative strategies for fighting multidrug resistant bacterial infections<\/a>. Biointerface Research In Applied Chemistry 9(1), 3834-3841<\/li>\n\n\n\n<li>Popa L.I., Czobor Barbu I., Chifiriuc M.C. 2018. Mobile genetic elements involved in the horizontal transfer of antibiotic resistance genes. Romanian Archives of Microbiology and Immunology, 77(4), 263-276.<\/li>\n\n\n\n<li>Vr\u00e2ncianu O., Chifiriuc M.C. 2018. Exploiting the bacterial adaptive immunity to develop new antisense \u2013 based strategies to combat antibiotic resistance. Romanian Archives of Microbiology and Immunology, 77 (1), 7-15.<\/li>\n<\/ul>\n\n\n\n<p class=\"wp-block-paragraph\"><em>Particip\u0103ri la conferin\u021be internationale<\/em><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Gheorghe I.; Czobor I; Avram I.; Alshaikhli N.; Pircalabioru-Gradisteanu G.; Negut A.C.; Marutescu L.; Lazar V.; Chifiriuc M.C. P0936. Genetic background of antibiotic resistance in multidrug-resistant Acinetobacter baumannii recovered from a wastewater treatment plant near Bucharest, Romania. 29th ECCMID, Amsterdam, Netherlands, from 13 &#8211; 16 Aprilie 2019.<\/li>\n\n\n\n<li>Cristea V.C., Angelescu M.C., Gheorghe I., Popa L.I., Czobor I., Ispas I., Grigore G.A., Bucatariu I., Marutescu L., Popa M., Lazar V., Popa I.M., Chifiriuc M.C. P1480. Snapshot of virulence and resistance markers in&nbsp;<em>Escherichia coli<\/em>&nbsp;uropathogenic strains isolated from ambulatory patients. 29th ECCMID, Amsterdam, Netherlands, from 13 &#8211; 16 April 2019.<\/li>\n\n\n\n<li>Czobor Barbu I., Popa L.I., Gheorghe I., S\u00e2rbu I., Negu\u021b A., Gr\u0103di\u0219teanu G., Laz\u0103r L., Chifiriuc M.C. Molecular characterization of multidrug resistant&nbsp;<em>Klebsiella pneumoniae<\/em>strains isolated from wastewater treatment plant samples in Romania. 29th ECCMID, Amsterdam, Netherlands, from 13 &#8211; 16 Aprilie 2019.<\/li>\n\n\n\n<li>Czobor Barbu I., S\u00e2rbu I., Gheorghe I., Popa L.I., Negu\u021b A., M\u0103ru\u021bescu L., Popa M., Gr\u0103di\u0219teanu G., Lazar V., Chifiriuc M.C. Detection of the epidemic IncP6 plasmid carrying&nbsp; blaKPC-2&nbsp; in a multidrug resistant&nbsp;&nbsp;<em>Enterobacter kobei<\/em>&nbsp;isolate from activated sludge of a wastewater treatment plant in V\u00e2lcea County, Romania. 29th ECCMID, Amsterdam, Netherlands, from 13 &#8211; 16 Aprilie 2019.<\/li>\n<\/ul>\n\n\n\n<h2 class=\"wp-block-heading has-vivid-green-cyan-background-color has-background has-medium-font-size\"><em>(EN)&nbsp;FINAL REPORT<\/em><\/h2>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>FINAL REPORT<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Project title&nbsp;<\/strong><em>In silico and experimental studies of mobile genetic platforms for multidrug resistance<\/em><\/li>\n\n\n\n<li><strong>Contract no. &#8211;&nbsp;<\/strong>PN-III-P3-3.1-PM-RO-CN-2018-0147 (CONTRACT NR. 5 BM)<\/li>\n\n\n\n<li><strong>Year of completion &#8211;&nbsp;<\/strong>2019<\/li>\n\n\n\n<li><strong>Duration of the project \u2013&nbsp;<\/strong>18 months<\/li>\n\n\n\n<li><strong>Romanian partner &#8211;&nbsp;<\/strong>University of Bucharest, Legal representative Rector prof.dr. Mircea Dumitru<\/li>\n\n\n\n<li><strong>Project Manager &#8211;&nbsp;<\/strong>univ. Dr. Mariana Carmen Chifiriuc<\/li>\n\n\n\n<li><strong>Foreign Partner &#8211;&nbsp;<\/strong>West China Hospital, Sichuan University, Project manager Prof. Ph.D. Xiaohui Wang<\/li>\n<\/ul>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; The general objective of the project is the investigation of local evolution of antibiotic resistance (AR) in multidrug-resistant (MDR) Gram-negative clinical isolates and from the wastewater network, by studying the genetic support of AR and the transferability of antibiotic resistance genes (ARGs) through mobile genetic elements (MGE), for a better understanding of the molecular epidemiology of AR in Romania (differentiation of locally emerging AR clones from regional \/ global ones) and the improvement of local management of AR.&nbsp;<strong>Scientific and technical description of the results and the degree of achievement of the objectives<\/strong><\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<strong>All proposed objectives and activities have been fulfilled<\/strong>.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;Within this project, there were analyzed 101 Gram-negative bacterial strains exhibiting the MDR phenotype (83 strains of&nbsp;<em>Klebsiella pneumoniae<\/em>, 12&nbsp;<em>Acinetobacter baumannii<\/em>, 3&nbsp;<em>Escherichia coli<\/em>, 1&nbsp;<em>Serratia<\/em>&nbsp;sp., 1&nbsp;<em>Enterobacter kobei<\/em>&nbsp;and 1&nbsp;<em>Providencia<\/em>&nbsp;sp.). The presence of carbapenem resistance genes as well as AR genes conferring ESBL phenotypes were investigated by simplex and multiplex PCR experiments.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; PCR experiments in&nbsp;<em>Enterobacteriaceae<\/em>&nbsp;isolates have shown the presence in decreasing order of the&nbsp;<em>bla<\/em><sub>SHV,&nbsp;<\/sub><em>bla<\/em><sub>NDM-1<\/sub>,&nbsp;<em>bla<\/em><sub>OXA-48<\/sub>,&nbsp;<em>bla<\/em><sub>TEM&nbsp;<\/sub>and&nbsp;<em>bla<\/em><sub>KPC<\/sub>&nbsp;genes. Regarding&nbsp;<em>A. baumannii<\/em>&nbsp;strains PCR experiments have shown the presence of the&nbsp;<em>bla<\/em><sub>OXA-51<\/sub>&nbsp;gene in all analyzed strains, followed by&nbsp;<em>bla<\/em><sub>OXA-23,<\/sub><em>bla<\/em><sub>SHV,&nbsp;<\/sub><em>bla<\/em><sub>VEB<\/sub>,&nbsp;<em>bla<\/em><sub>TEM,&nbsp;<\/sub><em>bla<\/em><sub>NDM-1&nbsp;<\/sub>and&nbsp;<em>bla<\/em><sub>VIM<\/sub>.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; The study of the AR transferability was performed by conjugation experiments carried out both in liquid medium (Liquid mating) and solid medium (Filter mating), for &gt;35 selected strains. The selection of transconjugants for&nbsp;<em>Enterobacteriaceae<\/em>&nbsp;was performed on both MacConkey medium supplemented with sodium azide (150 \u03bcg\/ml) and imipenem (1 \u03bcg\/ml), as selection agents and on BHI medium (Brain heart infusion) supplemented with sodium azide (150 \u03bcg\/ml) and imipenem (1 \u03bcg\/ml). In the case of&nbsp;<em>A. baumannii<\/em>&nbsp;strains, the selection was carried out on BHI medium supplemented with rifampicin (250 mg\/l) and meropenem (1 \u00b5g\/ml). Most of the&nbsp;<em>Enterobacteriaceae<\/em>&nbsp;strains strains included in the conjugation experiments and three of&nbsp;<em>A. baumannii<\/em>&nbsp;strains were found to be conjugative, most of them conjugating mainly on solid medium (filter). The PCR results obtained for the transconjugants have shown the transfer of some of the genes conferring \u03b2-lactam resistance, in the case of one&nbsp;<em>K. pneumoniae<\/em>, three genes being transferred simultaneously, suggesting the location of those genes in the same MGEs and the possibility of horizontal dissemination.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; The second phase of the project was dedicated to the sequencing of ARGs or bacterial genomes belonging to&nbsp;<em>K. pneumoniae<\/em>,&nbsp;<em>A. baumannii<\/em>,&nbsp;<em>E. coli<\/em>&nbsp;and&nbsp;<em>E. kobei<\/em>&nbsp;and the bioinformatics analysis, both of the sequencing data obtained, as well as of the conformation and interaction of the proteins encoded by the ARGs.&nbsp;<em>Next Generation Sequencing &#8211; pair-end whole genome sequencing &#8211; MiSeq System Sequencer, Illumina<\/em>&nbsp;followed by the bioinformatics analysis of sequencing products (using SPAdes, CARD, BLAST, ISFinder, PlasmidFinder, ResFinder, VirulenceFinder, INTEGRALL softwares) regarding the presence of ARGs have demonstrated that the analyzed strains harbored a high number of ARGs of antibiotic classes (between 4 and 8 classes). Regarding the presence of plasmids associated with AR for selected&nbsp;<em>Enterobacteriaceae<\/em>&nbsp;strains there have been revealed the presence of eight types of plasmids, namely ColE10, IncA\/C2, IncFIB (K), IncFII (Yp), IncHI1A (CIT), IncHI1B (CIT) ), IncP6, IncX1 in one&nbsp;<em>E. coli<\/em>&nbsp;strain, five Col440II, ColRNAI, IncFII (K), IncFII (Yp), IncR and three types, ColpVC, IncFII (Yp), IncL\/M (pOXA-48) plasmids respectively in&nbsp;<em>K. pneumoniae<\/em>&nbsp;strains. The three&nbsp;<em>Enterobacteriaceae<\/em>&nbsp;strains sequenced by NGS belonged to ST101 and ST383 clones (<em>K. pneumoniae<\/em>&nbsp;strains) and ST 378 (<em>E. coli<\/em>&nbsp;strain).<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; The conformational studies indicated conformations that the proteins OXA-23 and SHV-2 encoded by the ARG alleles identified by sequencing in the bacterial strains circulating in Romania were identical to those existing in the international databases.<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Project impact<\/strong><\/li>\n<\/ul>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; The collaboration with China (scientific visits, organization of international workshops) has essentially contributed to the development of human resources, offering the opportunity for the speecialization of Romanian team young members in&nbsp;<em>N<\/em><em>ext Generation Sequencing &#8211; pair-end whole genome sequencing &#8211; MiSeq System Sequencer, Illumina<\/em>&nbsp;and the bioinformatic analysis of sequencing products; this knowledge will be immediately apllied in other two international and one national ongoing research projects, granting the achievement of valuable results using state of the art methodology.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; Knowledge of the molecular epidemiology of AR in circulating Gram negative bacilli in Romania both in the clinical sector and in the aquatic environment (differentiation of locally emerging AR clones from regional \/ global ones) may have a major impact on improving the local management of AR.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; The obtained results are promising and will feature at least two major publications (one on&nbsp;<em>Klebsiella&nbsp;<\/em>ap. and one on&nbsp;<em>A. baumannii&nbsp;<\/em>strains) and three abstract that will be 3 abstracts will be submitted to the 30th European Congress of Clinical Microbiology and Infectious Diseases, Paris, France, 18-20 April 2020.<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>FINAL INDICATORS OF THE PROJECT<\/strong><\/li>\n\n\n\n<li><strong>Working visits during the bilateral project<\/strong><\/li>\n<\/ul>\n\n\n\n<p class=\"wp-block-paragraph\"><em>&nbsp; The visit of the Chinese team<\/em>&nbsp;(project manager: Wang Xiaohui, team members: Zong Zhiyong, Feng Yu, Liu Lina) in Romania: 31.05-11.06.2019<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; During the visit of the Chinese team at the Research Institute of the University of Bucharest (ICUB) there established the protocols to be followed within the project and also were selected the strains to be analyzed. Also, was carried out the training of the Romanian team by the Chinese partners regarding the conjugation experiments and selection of the transconjugants, but also on the bioinformatics analysis of the obtained data, respectively the assembly and the annotation and the detailed annotation of the bacterial genomes and their comparison.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><em>&nbsp; The visit of the Romanian team<\/em>&nbsp;(Research Assistant Ilda Barbu, Scientific Researcher Marius Surleac, PhD Laura Ioana Popa, PhD lecturer Irina Gheorghe) in China &#8211; Chengdu: 15-28.09.2019<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; During this visit, there were performed the bacterial conjugation experiments, selection of transconjugants and their analysis, sequencing of the selected bacterial genomes and bioinformatic data analysis.<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Specialization internships<\/strong><\/li>\n<\/ul>\n\n\n\n<p class=\"wp-block-paragraph\">&nbsp; The Romanian team (Research Assistant Ilda Barbu, Scientific Researcher Marius Surleac, PhD Laura Ioana Popa, PhD Irina Gheorghe) participated in a workshop organized by the host institution (Sichuan University, prof. Dr. Zong Zhiyong) regarding the bioinformatics of the bacterial genomes assembly and annotation.<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Dissemination of the results<\/strong><\/li>\n<\/ul>\n\n\n\n<p class=\"wp-block-paragraph\"><em>ISI papers<\/em><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Mitache M.M., Banu O., Borcan E., Gheorghe I., Marutescu M., Popa M., Predescu M., Rusu E., Nemes R., Chifiriuc M.C. 2018.&nbsp;<a href=\"http:\/\/scholar.google.com\/scholar?cluster=18418457897074986975&amp;hl=en&amp;oi=scholarr\">Resistance and Virulence Patterns in Gram Negative and Gram Positives Rods Isolated from the Hospital Environment in Bucharest, Romania<\/a>. Revista de Chimie 69 (11), 4026-4032<\/li>\n<\/ul>\n\n\n\n<p class=\"wp-block-paragraph\"><em>BDI papers<\/em><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Podgoreanu P., Negrea S., Buia R., Delcaru C., Trusca S.B., Lazar V., Chifiriuc M.C. 2019.&nbsp;<a href=\"http:\/\/scholar.google.com\/scholar?cluster=16293756831297448303&amp;hl=en&amp;oi=scholarr\">Alternative strategies for fighting multidrug resistant bacterial infections<\/a>. Biointerface Research In Applied Chemistry 9(1), 3834-3841<\/li>\n\n\n\n<li>Popa L.I., Czobor Barbu I., Chifiriuc M.C. 2018. Mobile genetic elements involved in the horizontal transfer of antibiotic resistance genes. Romanian Archives of Microbiology and Immunology, 77(4), 263-276.<\/li>\n\n\n\n<li>Vr\u00e2ncianu O., Chifiriuc M.C. 2018. Exploiting the bacterial adaptive immunity to develop new antisense \u2013 based strategies to combat antibiotic resistance. Romanian Archives of Microbiology and Immunology, 77 (1), 7-15.<\/li>\n<\/ul>\n\n\n\n<p class=\"wp-block-paragraph\"><em>International Conferences<\/em><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Gheorghe I.; Czobor I; Avram I.; Alshaikhli N.; Pircalabioru-Gradisteanu G.; Negut A.C.; Marutescu L.; Lazar V.; Chifiriuc M.C. P0936. 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 &amp; Infectious Diseases, Amsterdam, Netherlands, from 13 &#8211; 16 April 2019.<\/li>\n\n\n\n<li>Cristea V.C., Angelescu M.C., Gheorghe I., Popa L.I., Czobor I., Ispas I., Grigore G.A., Bucatariu I., Marutescu L., Popa M., Lazar V., Popa I.M., Chifiriuc M.C. P1480. Snapshot of virulence and resistance markers in&nbsp;<em>Escherichia coli<\/em>&nbsp;uropathogenic strains isolated from ambulatory patients. 29th European Congress of Clinical Microbiology &amp; Infectious Diseases, Amsterdam, Netherlands, from 13 &#8211; 16 April 2019.<\/li>\n\n\n\n<li>Czobor Barbu I., Popa L.I., Gheorghe I., S\u00e2rbu I., Negu\u021b A., Gr\u0103di\u0219teanu G., Laz\u0103r L., Chifiriuc M.C. Molecular characterization of multidrug resistant&nbsp;<em>Klebsiella pneumoniae<\/em>strains isolated from wastewater treatment plant samples in Romania. 29th European Congress of Clinical Microbiology &amp; Infectious Diseases, Amsterdam, Netherlands, from 13 &#8211; 16 April 2019.<\/li>\n\n\n\n<li>Czobor Barbu I., S\u00e2rbu I., Gheorghe I., Popa L.I., Negu\u021b A., M\u0103ru\u021bescu L., Popa M., Gr\u0103di\u0219teanu G., Lazar V., Chifiriuc M.C. Detection of the epidemic IncP6 plasmid carrying&nbsp; blaKPC-2&nbsp; in a multidrug resistant&nbsp;&nbsp;<em>Enterobacter kobei<\/em>&nbsp;isolate from activated sludge of a wastewater treatment plant in V\u00e2lcea County, Romania. 29th European Congress of Clinical Microbiology &amp; Infectious Diseases, Amsterdam, Netherlands, from 13 &#8211; 16 April 2019.<\/li>\n<\/ul>\n","protected":false},"excerpt":{"rendered":"<p>Studii in silico \u0219i experimentale ale unor platforme genetice mobile de rezisten\u021b\u0103 multipl\u0103 la antibiotice. (RO)&nbsp;RAPORT FINAL DE ACTIVITATE Titlul proiectului&nbsp;STUDII IN SILICO \u0218I EXPERIMENTALE [&hellip;]<\/p>\n","protected":false},"author":1,"featured_media":90,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[4],"tags":[],"class_list":["post-94","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-projects"],"_links":{"self":[{"href":"https:\/\/bios.unibuc.ro\/index.php?rest_route=\/wp\/v2\/posts\/94","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/bios.unibuc.ro\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/bios.unibuc.ro\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/bios.unibuc.ro\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/bios.unibuc.ro\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=94"}],"version-history":[{"count":9,"href":"https:\/\/bios.unibuc.ro\/index.php?rest_route=\/wp\/v2\/posts\/94\/revisions"}],"predecessor-version":[{"id":284,"href":"https:\/\/bios.unibuc.ro\/index.php?rest_route=\/wp\/v2\/posts\/94\/revisions\/284"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/bios.unibuc.ro\/index.php?rest_route=\/wp\/v2\/media\/90"}],"wp:attachment":[{"href":"https:\/\/bios.unibuc.ro\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=94"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/bios.unibuc.ro\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=94"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/bios.unibuc.ro\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=94"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}