Emergent multidrug resistant Escherichia coli increase clinical challenges. This thesis describes the resistance patterns, molecular epidemiology and mechanisms, for 315 E. coli from patients in the Vancouver Coastal Health Region for 2006/2007. Automated susceptibility testing was confirmed via E-test® for AmpC and/or ESBL production. PFGE, RFLP and PCR were used to assess genotypic relationships, and plasmid character. AmpC production was facilitated mainly by promoter mutations (54.5%). The principal ESBL detected was CTX-M-15 (49.5%). An unidentified ESBL-producer, with a pI near 8.3, was detected. A plasmid displayed variant resistance phenotypes dependent on selective growth media. A positive correlation between ST131 with CTX-M-15 and CIPR indicated the dissemination of companion phenotypes. Ciprofloxacin resistance resulted mainly (98.0%) from a double gyrA mutation. Overall fluoroquinolone resistance was not assessable due to exclusive selection parameters in this evaluation. Fluoroquinolone resistance factors require further examination to understand what causes resistant phenotypes exclusive of chromosomal mutations. ii

Emergent multidrug resistant Escherichia coli increase clinical challenges. This thesis describes the resistance patterns, molecular epidemiology and mechanisms, for 315 E. coli from patients in the Vancouver Coastal Health Region for 2006/2007. Automated susceptibility testing was confirmed via E-test® for AmpC and/or ESBL production. PFGE, RFLP and PCR were used to assess genotypic relationships, and plasmid character. AmpC production was facilitated mainly by promoter mutations (54.5%). The principal ESBL detected was CTX-M-15 (49.5%). An unidentified ESBL-producer, with a pI near 8.3, was detected. A plasmid displayed variant resistance phenotypes dependent on selective growth media. A positive correlation between ST131 with CTX-M-15 and CIPR indicated the dissemination of companion phenotypes. Ciprofloxacin resistance resulted mainly (98.0%) from a double gyrA mutation. Overall fluoroquinolone resistance was not assessable due to exclusive selection parameters in this evaluation. Fluoroquinolone resistance factors require further examination to understand what causes resistant phenotypes exclusive of chromosomal mutations. ii

BRAGA, J.M. Prevalência e sensibilidade antimicrobiana de escherichia coli e klebsiella pneumoniae isoladas de infecções nosocomiais no hospital regional norte em sobral/ce e detecção genética de blatem, blashv blactx-m e blages em espécimes produtores de betalactamase de espectro estendido (esbl). 2016. 101f. Dissertação (Mestrado em Ciências da Saúde) – Campus de Sobral, Universidade Federal do Ceará, Sobral, 2016. / Submitted by Mestrado Ciências da Saúde (ppgcsufcsobral@gmail.com) on 2017-05-25T14:15:01Z No. of bitstreams: 1 2016_dis_jmbraga.pdf: 4368780 bytes, checksum: f1881d9ef6d76e8cb665b910b9dcd076 (MD5) / Approved for entry into archive by Djeanne Costa (djeannecosta@gmail.com) on 2017-05-26T18:41:29Z (GMT) No. of bitstreams: 1 2016_dis_jmbraga.pdf: 4368780 bytes, checksum: f1881d9ef6d76e8cb665b910b9dcd076 (MD5) / Made available in DSpace on 2017-05-26T18:41:29Z (GMT). No. of bitstreams: 1 2016_dis_jmbraga.pdf: 4368780 bytes, checksum: f1881d9ef6d76e8cb665b910b9dcd076 (MD5) Previous issue date: 2016-12-01 / Escherichia coli and Klebsiella pneumoniae are Gram-negative bacilli account for a significant portion of infections in hospitals. Its importance has increased due to increased production of beta-lactamases of extended spectrum (ESBL). These enzymes are produced by some Gram negative bacilli and mediate resistance to beta-lactams, such as cephalosporins and aztreonam. In these pathogens, the majority of the ESBL identified are TEM, SHV and CTX-M types. This study aimed to analyze the prevalence and antimicrobial susceptibility of E. coli and K. pneumoniae isolated from nosocomial infections in North Regional Hospital in Sobral/CE, Brazil, from March 2015 to March 2016, and to detect blaTEM, blaSHV, blaCTX-M, blaKPC, blaVIM and blaGES genes of isolates which exhibited ESBL phenotype. A total of 245 isolates (132 E. coli and 113 K. pneumoniae) were analyzed. Of these, 145 (59.1%) had ESBL phenotype and 44 were characterized genetically by presence of blaTEM, blaSHV, blaCTX-M, blaKPC, blaVIM and blaGES genes. The rates of E.coli and Klebsiella pneumoniae ESBL producers were 49.2% and 70.8%, respectively. This research revealed that ESBL-producing E. coli strains were more sensitive to meropenem (100%), amikacin (96.9%), colistin (90.8%) and tigecycline (90.8%), and more resistant to ampicillin (100%), ceftriaxone (100%) and cefepime (96.9%). On the other hand, K. pneumoniae ESBL producers were more sensitive to colistin (100%), amikacin (96.2%) and meropenem (93.7%), and more resistant to ceftriaxone (100%), ceftazidime (100%), cefepime (98.7%) and ampicillin (98.7%). The blaCTX-M gene was detected in 14 (31.8%) isolates, blaTEM and blaSHV genes were detected both in 3 specimens (6.8%), the blaGES gene was detected in only one isolate (2.2%), while blaKPC and blaVIM genes were not detected. Our findings show high levels of resistance to beta-lactam antibiotics, as well as increasing linear trend for ESBL production by the studied species. The gene with the highest detection rates among the isolates was blaCTX-M gene, which is certainly the most important ESBL gene today. We detected one strain of Klebsiella pneumoniae harboring blaGES and blaCTX-M genes, being of our knowledge the first report in Brazil, which demonstrates the great potential for worldwide spread of resistance genes between Gram-negative bacillus. / Escherichia coli e Klebsiella pneumoniae são bacilos Gram-negativos responsáveis por uma parcela significante de infecções em ambiente hospitalar. Sua importância aumentou devido ao surgimento de espécimes produtoras de betalactamases de espectro estendido (ESBL). Essas enzimas são produzidas por alguns bacilos Gram-negativos e conferem resistência aos betalactâmicos, como cefalosporinas e aztreonam. Nesses patógenos, a maioria das ESBL identificadas é do tipo TEM, SHV e CTX-M. Este estudo teve como objetivo analisar a prevalência e a sensibilidade antimicrobiana de E.coli e K. pneumoniae isoladas de infecções nosocomiais do Hospital Regional Norte, Sobral/CE, Brasil no período de março de 2015 a março de 2016, assim como realizar a detecção dos genes blaTEM, blaSHV, blaCTX-M, blaKPC, blaVIM e blaGES nos isolados que exibiram fenótipo ESBL. No total, 245 isolados (132 E. coli e 113 K. pneumoniae) foram analisados. Destes, 145 (59,1%) apresentaram fenótipo ESBL e 44 foram caracterizados geneticamente quanto à detecção dos genes blaTEM, blaSHV, blaCTX-M, blaKPC, blaVIM e blaGES. As taxas de produção de ESBL por E.coli e K. pneumoniae foram 49,2% e 70,8%, respectivamente. Essa pesquisa revelou que E.coli produtoras de ESBL foram mais sensíveis ao meropenem (100%), amicacina (96,9%), colistina (90,8%) e tigeciclina (90,8%), e mais resistentes à ampicilina (100%), ceftriaxona (100%) e cefepima (96,9%). Já K. pneumoniae produtoras de ESBL foram mais sensíveis à colistina (100%), amicacina (96,2%) e meropenem (93,7%), e mais resistentes à ceftriaxona (100%), ceftazidima (100%), cefepima (98,7%) e ampicilina (98,7%). O gene blaCTX-M foi detectado em 12 (27,2%) isolados, os genes blaTEM e blaSHV foram detectados em 3 espécimes cada um (6,8%), o gene blaGES foi detectado em apenas um isolado (2,2%), enquanto os genes blaKPC e blaVIM não foram detectados. Nossos achados revelam altos índices de resistência aos betalactâmicos, assim como tendência linear crescente para produção de ESBL pelas espécies estudadas. O gene com maiores taxas de detecção entre os isolados foi o gene blaCTX-M que, certamente, é o gene ESBL mais importante clinicamente na atualidade. Nós detectamos um isolado de Klebsiella pneumoniae contendo o gene blaGES e blaCTX-M, sendo do nosso conhecimento o primeiro relato no Brasil, o que demonstra o grande potencial de disseminação mundial de genes de resistência entre bacilos Gram-negativos.

Orientador: Vera Lúcia Mores Rall / Resumo: A mastite bovina é uma das doenças que mais causam prejuízos às propriedades leiteiras e, apesar dos programas de controle, Escherichia coli destaca-se como importante patógeno ambiental causador desta enfermidade na sua forma clínica. Isolados obtidos de infecções nas mamas são classificadas como E. coli patogênica mamária (MPEC). O objetivo deste estudo foi caracterizar geneticamente isolados de E. coli a partir de leite de vacas com mastite clínica utilizando técnicas moleculares, bem como investigar os padrões de aderência e resistência aos antimicrobianos dos isolados, para isso a investigação dos grupos filogenéticos, genes de virulência, presença de clones (por Pulsed field gel electrophoresis, PFGE), tipos de adesão em células HeLa, resistência aos antimicrobianos e genes relacionados com essa resistência foram realizados em 110 E. coli isoladas de leite de vacas com mastite clínica de diferentes fazendas dos estados de São Paulo, Minas Gerais e Paraná. De acordo com a presença dos genes específicos arpA, chuA, yjaA e TspE4.C2, os isolados foram classificados principalmente nos grupos comensais A (50,9 %) e B1 (38,2 %), seguidos dos grupos D (2,7 %), C (1,8 %) e B2/E/F/Escherichia clade I (0,9 %). Três isolados (2,7 %) não foram categorizados em nenhum desses grupos, classificados como de grupo filogenético desconhecido. Nenhum dos 110 isolados apresentou genes que caracterizam E. coli diarreiogênicas (DEC), entretanto os genes astA e shf, geralmente encontrados em E.... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Bovine mastitis is one of the diseases that most causes damage to dairy properties and, despite control programs, Escherichia coli stands out as an important environmental pathogen that causes this disease in its clinical form. Isolates obtained from breast infections are classified as mammary pathogenic E. coli (MPEC). The objective of this study was to genetically characterize E. coli isolates from milk of cows with clinical mastitis using molecular techniques, as well as to investigate the adhesion patterns and antimicrobial resistance of isolates, for this purpose the investigation of phylogenetic groups, virulence genes, presence of clones (by Pulsed field gel electrophoresis, PFGE), HeLa cell adhesion types, antimicrobial resistance, and genes related to this resistance were performed on 110 E. coli isolated from milk of cows with clinical mastitis from different state farms from São Paulo, Minas Gerais and Paraná. According to the presence of the specific genes arpA, chuA, yjaA and TspE4.C2, isolates were classified mainly in commensal groups A (50.9 %) and B1 (38.2 %), followed by groups D (2.7 %), C (1.8 %) and B2/E/F/Escherichia clade I (0.9 %). Three isolates (2.7 %) were not categorized in any of these groups, classified as unknown phylogenetic group. None of the 110 isolates presented genes that characterize diarrheagenic E. coli (DEC), however, the astA and shf genes, commonly found in enteroaggregative E. coli, were identified in 7.3 % (8/110) and 10.3 % (3/29)... (Complete abstract click electronic access below) / Doutor

Bakgrund: Tarmbakterier med förmåga att bilda enzymet Extended Spectrum Beta Lactamase (ESBL) har varit känt inom sjukvården sedan 1980-talet. Intensivvårdssjuksköterskors erfarenhet av ESBL är lite utforskat samtidigt som ESBL utgör ett av de största hoten mot människors hälsa. Syfte: Syftet med studien var att undersöka intensivvårdssjuksköterskors erfarenheter vid vård av patienter med ESBL. Metod: Den studiedesign som användes var en intervjustudie med kvalitativ innehållsanalys. Semistrukturerade intervjufrågor ställdes till åtta intensivvårdssjuksköterskor från två universitetssjukhus i mellan Sverige. Resultat: Intensivvårdssjuksköterskornas erfarenhet av att vårda patienter med ESBL på en intensivvårdsavdelning beskrivs i resultatet genom fyra huvudkategorier: erfarenhet av ESBL, erfarenhet av rutiner kring smittsamma sjukdomar, erfarenheter vid svårigheter att följa hygienrutiner och erfarenhet att förmedla kunskap. Slutsats: Studien visar att de intervjuade intensivvårdssjuksköterskorna har erfarenhet av att vårda en patient med ESBL på intensivvårdsavdelningar och är skickliga i sitt yrke enligt Benners teori Förslag på vidare forskning: Denna studie skulle kunna ligga till grund för enkätstudie om intensivvårdssjuksköterskans kunskap, kompetens, attityder och upplevda hinder för att bedriva vård enligt riktlinjer och föreskrifter. / Background: Intestinal bacteria capable of producing the enzyme Extended Spectrum Beta Lactamase (ESBL) has been known in health care since the 1980s. There is limited research about the intensive care nurses experience of ESBL while ESBL represents one of the greatest threats to human health. Purpose: The aim of the study was to examine intensive care nurses' experiences of care for patients with ESBL. Method: The study design used was an interview study with qualitative content analysis. Semi-structured interview questions were asked to eight intensive care nurses from two university hospitals in middle part of Sweden. Results: Four main categories were identified: experience of ESBL, experience of routines for infectious diseases, experience the difficulty to follow hygiene and experience to impart knowledge. Conclusion: Intensive care nurse have experience of caring for a patient with ESBL in intensive care and are proficient according to Benners theory. Suggestions for further research: This study could form the basis for the survey study of intensive care nurses' knowledge, skills, attitudes and perceived obstacles to providing care according to the guidelines and regulations.

O objetivo deste estudo foi caracterizar bactérias multirresistentes (MDR) isoladas de cavalos saudáveis (fezes) e doentes no Brasil e na França. De março de 2012 a dezembro de 2014, amostras clínicas coletadas de cavalos saudáveis e doentes no Brasil foram selecionadas para pesquisa da presença de bactérias MDR. A investigação sobre as amostras franceses foi restria a isolados de Escherichia coli (EC) recuperados a partir de amostras clínicas coletadas entre 2014 e 2015. Nos cavalos brasileiros, a análise de amostras de fezes de animais saudáveis revelou a presença de clones de EC não relacionados pertencentes aos filogrupos A, D ou B2 que carreavam genes como: blaCTX-M-1, blaCMY-2, qnr- e genes add-tipo (amino-transferases); enquanto que nos cavalos doentes foram encontradas EC, Proteus mirabilis, Klebsiella pneumoniae, Pseudomonas aeruginosa e Serratia marcescens carreando genes blaCTX-M-15, blaCTX-M-1, rmtD 16S rRNA metilase, qnr-tipo, aac(6´)-Ib-cr e aad-tipo. Nos cavalos doentes franceses de EC MDR foram positivas para CTX-M-1, seguido de M-2- e M-9. Estes resultados destacam a importância de cavalos como um novo reservatório de bactérias MDR. / The aim of this study was to characterize multidrug-resistant (MDR) bacteria isolated from healthy and infected horses in Brazil and France. From March 2012 to December 2014, clinical samples collected from healthy and infected horses, in Brazil, were screened for the presence of MDR bacteria. Investigation on French isolates was restricted to E. coli strains recovered from clinical samples collected between 2014 and 2015. In Brazilian horses, the analysis of fecal samples from healthy animals revealed the presence of clonally unrelated A, D or B2 phylogroups of E. coli strains carrying blaCTX-M-1, blaCMY-2, qnr- and aminoglycoside adenyl transferase (aad)-type genes, whereas in infected horses, E. coli, Proteus mirabilis, Klebsiella pneumoniae, Pseudomonas aeruginosa and Serratia marcescens isolates carrying blaCTX-M-15, blaCTX-M-1, rmtD 16S rRNA methylase, qnr-type, aac(6´)-Ib-cr and aad-type genes. In French infected horses, most MDR E. coli isolates were positive for CTX-M-1-, followed by CTX-M-2- and CTX-M-9-type extended-spectrum beta-lactamases. These results highlight the importance of horses as a new reservoir of MDR bacteria.

Animais de companhia têm sido apontados como reservatórios de bactérias gram-negativas resistentes a antibióticos utilizados em medicina humana e veterinária. O objetivo deste estudo foi investigar a prevalência da resistência mediada por plasmídeos em bactérias Gram-negativas isoladas de animais de companhia no Brasil e na França, elucidando o papel potencial desses animais como portadores assintomáticos. Amostras de DNA extraídas de quatro coleções de bactérias Gram-negativas produtoras de ESBL foram analisadas por tipagem e sub-tipagem baseados em PCR, análise do polimorfismo de comprimento de fragmentos de restrição (RFLP), dimensionamento baseado em PFGE de nuclease S1 e hibridação Southern blot. Adicionalmente, isolados de Escherichia coli, Klebsiella pneumoniae e Enterobacter cloacae foram caracterizados por PFGE (Pulsed-field Gel Electrophoresis) e Multilocus Sequence Typing, agrupamento filogenético e tipagem de O25b. A presença de plasmídeos IncH12 (~600-kb) e IncFIB (~210-290-kp) carregando genes blaCTX-M-15 e blaCTX-M-9, foi confirmada entre cepas de E. coli isoladas de animais brasileiros, enquanto uma predominância de plasmídeos IncI1 (~200 kb) pertencentes ao complexo clonal (CC) CC12 contendo o gene blaCMY-2 foi observado entre linhagens de E. coli, em filogrupos de baixa virulência A e B1. A presença de plasmídeos do tipo IncHI2 (~ 600kb) carregando o gene blaCTX-M-15 foi confirmada em cepas de E. cloacae ST927 isoladas de fezes e saliva de cães assintomáticos no Brasil. Entre os animais franceses com infecções, os isolados de E. coli pertencentes ao filogrupo A, B1 e B2 apresentaram tamanho de plasmídeo IncF de ~210-290 kb, carregando principalmente genes blaCTX-M-15, além da presença de plasmídeo IncI1 carregando em sua maioria genes blaCTX-M-1, blaCTX-M-9 e blaCMY-2. Em animais franceses saudáveis, além das associações blaCTX-M-15/IncI1, blaCTX-M-1/IncFIB, blaCTX-M-14/IncF e da presença de blaCMY-2 e blaTEM-52b (não tipáveis), foi identificada uma cepa de E. coli carregando um plasmídeo IncL (~60kb) contendo o gene blaOXA-48, sendo esta a primeira descrição desse gene em animais na França. Além disso, os genes blaCTX-M-15, blaCTX-M-2 e blaCTX-M-9 foram localizados no cromossomo em cepas brasileiras e francesas, como observado por Southern blot e Sequenciamento de Nova Geração (NGS). Em resumo, em ambos os países, a prevalência de cepas positivas para a resistência tipo ESBL é grande. Os animais de companhia podem ter um papel importante na disseminação dos genes AmpC e ESBL mediados por plasmídeos. / Companion animals can be reservoirs of Gram-negative bacteria resistant to antibiotics used in human and veterinary medicine. The aim of this study was to investigate the genetic context of plasmid-mediated resistance in Gram-negative bacteria isolated from companion animals in Brazil and France, elucidating the potential role of these animals as asymptomatic carriers. DNA samples, extracted from a collection of ESBL-producing Gram-negative bacteria, were analyzed by PCR-based typing and sub-typing schemes, restriction fragment length polymorphism analysis, S1 nuclease PFGE-based sizing and Southern blot hybridization. Additionally, Escherichia coli, Klebsiella pneumoniae, and Enterobacter cloacae isolates were characterized by PFGE (Pulsed-field Gel Electrophoresis), Multilocus Sequence Typing, phylogenetic grouping and O25b typing. Presence of IncH12 (~600-kb) and IncFIB (~210-290-kp) plasmids carrying blaCTX-M-15 and blaCTX-M-9, respectively, was confirmed among E. coli strains isolated from Brazilian pets, whereas predominance of IncI1 plasmids (~200 kb) belonging to the clonal complex (CC) CC12 and carrying blaCMY-2 gene was observed among E. coli strains of low-virulence phylogrups A and B1. The presence of IncHI2-type (~ 600-kb) plasmids carrying blaCTX-M-15 gene was confirmed in E. cloacae strains ST927 isolated from feces and saliva from asymptomatic dogs in Brazil. Among French diseased companion animals, E. coli isolates belonging to phylogroup A, B1 and B2 were found carrying IncF-type plasmid with size of ~210-290-kb, which harbored mainly blaCTX-M-15 genes. In addition, presence of IncI1 plasmids carrying blaCTX-M-1, blaCTX-M-9 and blaCMY-2 genes was identified. In healthy French animals, besides associations blaCTX-M-15/IncI1, blaCTX-M-1/IncFIB, blaCTX-M-14/IncF, and the presence of blaCMY-2 and blaTEM-52b (non-typable), it was observed an E. coli strain carrying an IncL plasmid (~ 60kbp) containing the blaOXA-48 gene, representing the first description of this gene in a French dog. In addition, blaCTX-M-15, blaCTX-M-2 and blaCTX-M-9 genes were located on the chromosome in Brazilian and French strains, as observed by Southern Blot and New Generation Sequencing (NGS) analysis. In summary, in both countries, the prevalence of positive strains for ESBL-type resistance in cats and dogs is high. Companion animals may play an important role in the dissemination of the plasmid mediated AmpC and ESBL genes.

Bakgrund: ESBL är ett enzym som kan produceras av bakterier i tarmens normalflora och gör bakterien motståndskraftig –resistent- mot många antibiotika. För den enskilde individen får det konsekvenser vid en infektion orsakad av ESBL-bildande tarmbakterier, och då behandling krävs. De som infekteras med dessa bakterier riskerar att svara dåligt på behandling med våra vanligaste antibiotika, och det kan krävas inläggning på sjukhus även vid banala infektioner. Syfte: Att fördjupa kunskapen om vad det innebär för den enskilda individen att drabbas av ESBL-bildande tarmbakterier. Metod. En modifierad variant av Grounded Theory användes som analysmetod av sju öppna intervjuer. Resultat: I analysen växte kärnkategorin Att bli utkastad i det skrämmande och okända utan karta och kompass fram. Samtliga informanter upplevde att de fått bristande information, eller ingen information alls om sin diagnos. Informationsvägen från läkaren var antingen via telefon eller genom ett brev via posten. Konsekvensen blev att det uppstod många tankar och funderingar efteråt, och det innefattade även frågan om hur man blivit smittad; genom sjukvården eller om de själva orsakat att bli smittade. I mötet med sjukvården upplevdes att okunskapen hos personalen i vissa fall orsakade stigmatisering. Här förekom såväl extrema hygienåtgärder i form av “skyddsmundering“ som alltför bristande hygienrutiner. Därutöver framkom också upplevda attitydproblem från personalens sida, nonchalans, bristande förståelse, ingen villighet eller tid att svara på frågor. Allt detta ledde till att informanterna i sin egen vardag fick ta saken i egna händer. Samtliga försökte skaffa sig information på annat sätt, exempelvis via internet. I oron för att smitta andra konstruerades egna åtgärder av kvinnorna i studien, exempelvis att instruera andra om handtvätt, att själv desinfektera föremål vid vårdbesök och i bostaden, att inte åka med tunnelbana eller buss, att inte umgås med andra och att själv uppleva informationsplikt om sin smitta. Männen i studien vidtog, trots bristande eller ingen information alls, inte några speciella åtgärder i sitt vardagsliv, de fortsatte att leva som tidigare. Ingen ville oroa sina anhöriga/närstående. Det förekom att barnen ej informerats alls om diagnosen. Konklusion: För att kunna hantera sin livssituation är det av stor betydelse att den som drabbas av en ESBL-bildande bakterie får en god information av patientansvarig läkare. / Background: Extended spectrum beta lactamase (ESBL), an enzyme produced by bacteria in normal intestinal flora, renders such bacteria resistant to many antibiotics. Some patients infected with ESBL respond poorly to antibiotic treatment, and even trivial infections may require hospitalization.Purpose: To increase understanding of the emotional impact of ESBL-producing intestinal bacteria.Method: This study used a modified version of grounded theory during seven open interviews to analyze coping mechanisms for ESBL infection.Results: Our analysis identified a core category (i.e., being thrown into scary and unknown territory without a map and compass). All respondents felt they received no or insufficient information about the diagnosis, and reported that any information they did receive arrived only by phone or letter. Consequently, respondents questioned whether they had been infected through medical care or through their own actions. They believed that lack of knowledge and attitude problems among healthcare providers (perceived as carelessness, lack of understanding, and unwillingness or lack of time to answer questions) stigmatizes patients. Such deficits led respondents to take matters into their own hands as they tried to obtain information by other means (e.g., the Internet). Respondents described extreme hygiene measures as a "protective suit" against inadequate hygiene. Female respondents constructed individual coping mechanisms (e.g., instructing others about hand washing technique, disinfecting objects during healthcare visits and at home, avoiding metro or bus travel, avoiding social interactions, and informing others of the infection). Conversely, male respondents took no special measures and lived as they did before infection. No one wanted to worry relatives/significant others, and no one told their children about the diagnosis.Conclusion: It is to important that attending doctors provide good information to individuals infected by ESBL-producing bacteria. Moreover, such individuals must develop good life management and coping skills. / <p>ISBN 978-91-86739-12-6</p>

Der zeitnahe und möglichst sichere Nachweis bakterieller Krankheitserreger und deren Empfindlichkeit gegenüber verfügbaren antibakteriell wirksamen Chemotherapeutika (Antibiotika) stellt einen Hauptaufgabenbereich der medizinischen mikrobiologischen Routinediagnostik dar. Hierzu wurden im Laufe der Jahre unterschiedliche Methoden entwickelt, womit von der genauen Beschreibung der Kolonie- und mikroskopischen Morphologie, Anfärbbarkeit und Formation über die Charakterisierung der biochemischen Leistungsfähigkeit bis hin zur genauen Sequenzierung des gesamten Genoms ein enormer Fortschritt zu verzeichnen war. Seit Mitte der 1990er Jahre etablierte sich die Massenspektrometrie als phänotypisches Nachweisverfahren und gewann zunehmend an Bedeutung. Ebenso konnten Erfolge beim Nachweis Antibiotika resistenter Bakterien verzeichnet werden. Um das Potential dieser noch jungen Nachweismethode weiter zu erforschen, wurden in dieser Arbeit Spezies der Familie Enterobacteriaceae und der Nonfermenter in eine eigene massenspektrometrische Datenbank aufgenommen, um diese als Grundlage zur Validierung des Identifizierungspotentials der Methode mittels Blindstudie zu nutzen. Im selben Arbeitsschritt wurde der Versuch unternommen, Antibiotika resistente Stämme im Zuge der Speziesidentifizierung zu detektieren, um so Aussagen über eine mögliche Einschränkung der therapeutischen Möglichkeiten und gegebenenfalls notwendigen Hygienemaßnahmen treffen zu können.

O objetivo deste estudo foi caracterizar bactérias multirresistentes (MDR) isoladas de cavalos saudáveis (fezes) e doentes no Brasil e na França. De março de 2012 a dezembro de 2014, amostras clínicas coletadas de cavalos saudáveis e doentes no Brasil foram selecionadas para pesquisa da presença de bactérias MDR. A investigação sobre as amostras franceses foi restria a isolados de Escherichia coli (EC) recuperados a partir de amostras clínicas coletadas entre 2014 e 2015. Nos cavalos brasileiros, a análise de amostras de fezes de animais saudáveis revelou a presença de clones de EC não relacionados pertencentes aos filogrupos A, D ou B2 que carreavam genes como: blaCTX-M-1, blaCMY-2, qnr- e genes add-tipo (amino-transferases); enquanto que nos cavalos doentes foram encontradas EC, Proteus mirabilis, Klebsiella pneumoniae, Pseudomonas aeruginosa e Serratia marcescens carreando genes blaCTX-M-15, blaCTX-M-1, rmtD 16S rRNA metilase, qnr-tipo, aac(6´)-Ib-cr e aad-tipo. Nos cavalos doentes franceses de EC MDR foram positivas para CTX-M-1, seguido de M-2- e M-9. Estes resultados destacam a importância de cavalos como um novo reservatório de bactérias MDR. / The aim of this study was to characterize multidrug-resistant (MDR) bacteria isolated from healthy and infected horses in Brazil and France. From March 2012 to December 2014, clinical samples collected from healthy and infected horses, in Brazil, were screened for the presence of MDR bacteria. Investigation on French isolates was restricted to E. coli strains recovered from clinical samples collected between 2014 and 2015. In Brazilian horses, the analysis of fecal samples from healthy animals revealed the presence of clonally unrelated A, D or B2 phylogroups of E. coli strains carrying blaCTX-M-1, blaCMY-2, qnr- and aminoglycoside adenyl transferase (aad)-type genes, whereas in infected horses, E. coli, Proteus mirabilis, Klebsiella pneumoniae, Pseudomonas aeruginosa and Serratia marcescens isolates carrying blaCTX-M-15, blaCTX-M-1, rmtD 16S rRNA methylase, qnr-type, aac(6´)-Ib-cr and aad-type genes. In French infected horses, most MDR E. coli isolates were positive for CTX-M-1-, followed by CTX-M-2- and CTX-M-9-type extended-spectrum beta-lactamases. These results highlight the importance of horses as a new reservoir of MDR bacteria.