Introduction: In Sweden, approximately 57,000 people suffer from Healthcare associated infection (HAI) annually, of which half of the infections are estimated to be avoidable. This can lead to consequences such as extended treatment time, increased costs for the patient and increased patient mortality as well as increased care costs. Today, there is a large scientific base on prevention and the cause of HAI from a healthcare perspective. However, there is a lack of evidence regarding patients' experience of having suffered a HAI. Attention to patients' experiences contributes to increased knowledge among healthcare personnel, which can enable improved care that can be adapted to the patient's needs. Aim: The aim of the literature study was to investigate patients' experiences of being affected by a healthcare-associated infection. Method: The study has a literature study as a method with a descriptive design and a qualitative approach, which was based on eleven original scientific articles. Result: The analysis of the articles resulted in two categories with four subcategories each. The category An inner perspective of HAI described patients' experiences of HAI on a personal level, where patient experiences of being affected, isolated, limitation, impact on social relationships, new routines and behaviors were identified. The category An external perspective of HAI describes patients' experience of HAI influenced by factors from the environment where experiences in information, communication, care management, finances and the accusation have been identified. Conclusion: Many and different dimensions of experiences emerged throughout the HAI course that affected the quality of life physically, emotionally and socially. Patients' experiences could be traced to various causes that could have been addressed. The result indicated a lack of care management in the form of substandard communication, information, knowledge and hygiene routine. Also a lack of care management regarding person-related care with a lack of awareness that patients had emotional experiences and how these could be responded to. Keywords: Literature study, nursing, patient perspective, healthcare-associated infection, care science / Introduktion: I Sverige drabbas cirka 57 000 personer årligen av en vårdrelaterad infektion (VRI) varav hälften av infektionerna uppskattningsvis kan undvikas. Det kan leda till konsekvenser som förlängd vårdtid, ökade kostnader för patienten och ökad patientdödlighet samt ökade vårdkostnader. Det finns idag ett stort vetenskapligt underlag om prevention och orsak till VRI ur ett vårdperspektiv. Däremot förekommer bristande underlag om patienters upplevelse av att ha drabbats av VRI. Uppmärksamhet av patienters upplevelser och erfarenheter bidrar till ökad kunskap hos vårdpersonal, vilket kan möjliggöra förbättrat omhändertagande och förbättrande åtgärder som kan anpassas till patienternas behov. Syfte: Syftet med litteraturstudien var att undersöka patienters upplevelser av att drabbas av en vårdrelaterad infektion Metod: Studien har litteraturstudie som metod med deskriptiv design och kvalitativ ansats, som baserades på elva vetenskapliga orginalartiklar. Resultat: Analysen av artiklarna resulterade i två kategorier med fyra subkategorier vardera. Kategorin Ett inre perspektiv av VRI beskrev patienters upplevelser av VRI på det personliga planet där patientupplevelser av att drabbas, isolering, begränsning, påverkan på sociala relationer samt nya rutiner och beteenden identifierats. Kategorin Ett yttre perspektiv av VRI beskriver patienters upplevelse av VRI påverkat av faktorer från omgivningen där upplevelser inom information, kommunikation, vårdhantering, ekonomi samt ansvarsfrågan identifierats.  Slutsats: Det framkom många och olika dimensioner av upplevelser genom hela VRI- förloppet, som påverkat livskvaliteten fysiskt, emotionellt och socialt. Patienters upplevelser kunde härledas till olika orsaker som hade kunnat åtgärdas. Resultatet visade på bristande vårdhantering i form av undermålig kommunikation, information, kunskap och hygienrutin. Även bristande vårdhantering gällande personcentrerad vård med avsaknad av en medvetenhet kring att patienter hade emotionella upplevelser och hur dessa kunde bemötas. Nyckelord: Litteraturstudie, omvårdnad, patientperspektiv, vårdrelaterad infektion, vårdvetenskap

En France, les infections associées aux soins concernent environ un patient hospitalisé sur vingt. Les pathogènes en cause se transmettent d’unepersonne à l’autre par contact direct entre les personnes mais aussi par les surfaces de contact sur lesquelles certains microorganismes peuventpersister jusqu’à plusieurs mois. Le cuivre étant un puissant antimicrobien naturel, des éléments en alliages de cuivre ont été conçu. Ce travailde thèse vise à étudier la capacité de ces éléments à réduire les contaminations bactériennes lors d’une utilisation prolongée en établissementsde santé. Pour cela, cinq EHPAD et une MARPA (Marne, France) ont été équipées à 50% de poignées de portes et rampes de maintien en alliagesde cuivre. Plus de 1300 prélèvements bactériologiques ont été effectués sur la surface des éléments en établissements entre 1,5 et 3,5 ans aprèsleur installation. Les bactéries récoltées ont été cultivées sur différents milieux gélosés et les unités formant colonie ont pu être dénombrées. Ledénombrement a révélé que les niveaux de contamination sont significativement plus faibles sur les poignées et rampes en cuivre que sur leséléments standards. L’identification des souches bactériennes récoltées a ensuite été effectuée par spectrométrie de masse MALDI-TOF. Cetteanalyse a montré que les genres prédominants sur les surfaces de contact sont Staphylococcus et Micrococcus, et que les genres Staphylococcus,Streptococcus et Roseomonas sont significativement moins fréquents sur les éléments en cuivre. L’espèce pathogène. S. aureus a été observémoins fréquemment sur les éléments en cuivre que sur les éléments standard. Les éléments en alliage de cuivre sont donc efficaces pour éviterdes contaminations bactériennes de surfaces en milieux de santé. De plus, les propriétés antibactériennes des éléments en alliages de cuivresont conservées plusieurs années après leur mise en service, soulignant l’intérêt de leur utilisation en milieu de santé. / In France, healthcare-associated infections concern one on twenty patients during hospitalization. Pathogenic microorganisms spread from oneperson to another by direct contact between people, but also through touch surfaces where the can persist up to several months. Copper is anatural and powerful antimicrobial metal. Thus, copper alloyed elements and surfaces have been designed and manufactured. This thesis workaims to investigate on the ability of copper elements to reduce bacterial contaminations during an extended period of use in healthcare facilities.Five long-term care facilities were 50% outfitted with copper alloyed door handles and handrails. Over 1300 samplings were performed between1.5 and 3.5 years after copper elements installation. Sampled bacteria were cultivated on a range of agar plates and colony forming units werecounted. It revealed that contamination levels were lower on copper door handles and handrails than on controls. Identifications of sampledbacterial strains were then performed by MALDI-TOF mass spectrometry. This analysis showed that Staphylococcus and Micrococcus largelyprevailed on touch surfaces and that Staphylococcus, Streptococcus and Roseomonas are significantly less frequent on copper elements surfaces.Pathogenic species S. aureus was less frequently observed on copper elements than on controls. This study suggests that copper alloyedelements are effective to limit bacterial contaminations of surfaces in healthcare facilities. Moreover, these elements still display significativeantibacterial properties after several years of use. Thus, copper alloyed elements represent a very promising solution to control bacterialcontamination of touch surfaces in healthcare facilities.

Introduktion: Vårdrelaterade infektioner och antibiotikaresistens är ett folkhälsoproblem. Kvarliggande urinvägskateter är en riskfaktor för att drabbas av vårdrelaterad urinvägsinfektion. Det finns en tydlig koppling mellan vårdrelaterad infektion, antibiotikaanvändning och antibiotikaresistens. På sjukhusronden har sjuksköterskan en nyckelroll i att förebygga vårdrelaterade infektioner. Syfte: Undersöka om införandet av ett rondkort har någon effekt på sjuksköterskornas upplevelse av sin delaktighet i patientens antibiotika- och urinvägskateterbehandling. Metod: En interventionsstudie utfördes på två vårdavdelningar på Norrlands universitetssjukhus, Umeå, Västerbotten. Interventionen bestod av en checklista, RondkortVLL, som utvecklades för sjuksköterskor att använda vid ronden. Interventionen pågick under fyra veckor. För att mäta deras upplevelse av delaktighet i patienternas antibiotika- och KAD-behandling fick de svara på en enkät före och efter interventionen. Svaren före och efter interventionen jämfördes med Mann Whitney-U test. Resultat: En signifikant förbättring på 1) dagligt ställningstagande till KAD-behandling och 2) sjuksköterskorna upplevde det naturligt att föra en dialog med läkaren om alternativ till urinvägskateter med mindre infektionsrisk. En ökad upplevelse av delaktighet angående patienternas antibiotikabehandling kunde ses på svaren efter interventionen men denna ökning var inte signifikant. Slutsats: Eftersom urvalet var litet och endast två avdelningar undersöktes är det svårt att dra några generella slutsatser. Resultatet tyder på att RondkortVLL har ökat sjuksköterskornas upplevelse av delaktighet i patienternas KAD-behandling och kan ha ökat sjuksköterskornas upplevelse av delaktighet i patienternas antibiotikabehandling. På grund av resultaten och betydelsen av att förebygga vårdrelaterade infektioner förordar vi en ökad klinisk användning av RondkortVLL. Vi hävdar att RondkortVLL kan vara effektivt som en daglig påminnelse, stärka sjuksköterskorna och medvetandegöra dem om deras betydelse i alla aspekter i patientens vård. / Introduction: Healthcare-associated infections and antibiotic resistance are public health problems. Indwelling catheters increase risk for urinary tract infections. There are connections among healthcare-associated infections, antibiotic use and antibiotic resistance. Nurses play a key role in infection control during hospital ward rounds. Aim: To investigate if a ward round checklist affects nursesʼ experiences of participation in patients’ antibiotica and indwelling catheter treatments. Method: An intervention study was conducted in two wards at Norrland University Hospital. The intervention was use of a checklist, RondkortVLL, that we developed for use by nurses at the ward round. The duration of the intervention was 1 month. The nurses answered a questionnaire about the use of antibiotica- and indwelling catheter before and after the intervention. Pre- and post-intervention responses were compared using the Mann Whitney-U test. Results: Significant improvements were found in 1) daily consideration of indwelling catheters and 2) the nurses feeling it was natural to conduct a dialogue with the physician about lower infection risk alternatives to indwelling catheters. While nurses tended to more often report the experience of participation in antibiotic treatment considerations post-intervention, this was not significant. Conclusion: Due to a small sample size and only two wards investigated it is difficult to generalize our results. However, our data indicate that RondkortVLL improved the nurses' experience of participation in patients' indwelling catheter treatments and may improve nursesʼ experiences of participation in a patient’s antibiotic treatment. Given these findings and the importance of infection prevention and control, we are pursuing greater clinical utilization of the RondkortVLL. We assert that RondkortVLL can be useful as a daily reminder, empower the nursesʼand raise awareness of their importance in all aspects of patient care.

Abstract Healthcare-associated infections (HAI) are infections acquired during treatment in a healthcare facility. The most common infections are pneumonias, surgical site infections (SSIs) and urinary tract infections (UTIs). HAIs burden the healthcare system by increasing patient days, the use of antibiotics, examinations, and thus the costs of care. The occurrence of HAIs can be used to evaluate the quality of care and to make comparisons between institutions. The purpose of this dissertation was to evaluate the burden of HAIs in the primary and tertiary healthcare wards and the costs of procedure-related prosthetic joint infections (PJIs). The first part of this dissertation evaluated the prevalence of HAIs in the wards of primary healthcare in the Oulu University Hospital (OUH) district with two point prevalence studies (one-day sampling) conducted in 2006 and in 2017. In 2006, the study comprised 27 healthcare centres with 44 wards and 1,294 patients. HAIs were found in 9.3% of the patients. The most common infections were UTIs, skin and soft tissue infections and lower respiratory tract infections (LRTIs). In 2017, there were 20 healthcare centres with 34 wards and 764 patients; 9.4% of the patients had a HAI. The most common HAIs were pneumonias, SSIs and LRTIs. In the second part, we evaluated the incidence of HAIs in the OUH with a computer-based electronic infection surveillance program. The study covered 15 adult wards with a total of 353 beds. The overall incidence of HAIs during the six-year study period was 4.5% of discharged patients. The most common infections were SSIs, pneumonias and UTIs. The surveillance carried out in this way required a total of one person's workload per year. The third part evaluated hospital costs of procedure-related PJIs. The study population consisted of all total knee and hip arthroplasties performed in the OUH from 2013 to 2015: 1,768 patients with 42 PJIs. A PJI tripled the cost of a procedure compared to an arthroplasty without an infection (€25,100 vs. €7,200). Two-stage revision caused three times more costs than debridement, antibiotics and implant retention treatment (DAIR) (€53,400 vs. €18,500). HAIs are common in the wards of primary and tertiary healthcare in the OUH district. Electronic HAI monitoring is feasible but requires relatively large employer resources. Postoperative PJI triples the cost of the procedure. / Tiivistelmä Sairaalainfektio (SI) on infektio, jonka potilas saa ollessaan hoidossa laitoksessa. Yleisimpiä SI:ita ovat leikkausalueen infektio, keuhkokuume ja virtsatieinfektio (VTI). SI:t kuormittavat terveydenhoitoa lisäämällä hoitopäiviä, antibioottien käyttöä, tutkimuksia ja näin myös hoidon kustannuksia. SI:iden määrää voidaan käyttää hoidon laadun mittarina sekä sairaaloiden väliseen vertailuun. Tämän väitöskirjan tarkoituksena oli arvioida SI:iden määrää Pohjois-Pohjanmaan sairaanhoitopiirin (PPSHP) terveyskeskusten ja Oulun yliopistosairaalan (OYS) vuodeosastoilla sekä selvittää tekonivelleikkauksen jälkeisen tekonivelinfektion (TI) aiheuttamia sairaalakustannuksia. Väitöskirjan ensimmäisessä osatyössä selvitettiin SI:iden esiintyvyyttä PPSHP:n terveyskeskusten vuodeosastoilla pisteprevalenssilla eli yhden päivän otannalla vuosina 2006 ja 2017. Vuoden 2006 tutkimuksessa oli 27 terveyskeskusta, joissa oli 44 vuodeosastoa ja yhteensä 1 294 potilasta. SI todettiin 9,3 %:lla potilaista. Yleisimpiä olivat VTI:t, pehmytkudosinfektiot ja alahengitystieinfektiot. Vuonna 2017 tutkimuksessa oli 20 terveyskeskusta, 34 vuodeosastoa ja 764 potilasta. Tällöin SI todettiin 9,4 %:lla. Yleisimmät infektiot olivat keuhkokuume, leikkausalueen infektio ja alahengitystieinfektio. Toisessa osatyössä selvitettiin OYS:n SI:iden ilmaantuvuutta kuuden vuoden ajan jatkuvan infektioseurannan mahdollistavan tietokoneohjelman avulla. Tutkimuksessa seurattiin 15:ttä aikuisvuodeosastoa, joissa oli yhteensä 353 potilaspaikkaa. SI todettiin 4,5 %:ssa hoitojaksoista. Seuranta vaati yhteenlaskettuna noin yhden hoitajan työpanoksen vuodessa. Väitöskirjan kolmannessa osatyössä selvitettiin vuosina 2013–2015 OYS:ssa tehtyjen tekonivelleikkausten jälkeisten TI:iden sairaalakustannuksia. Tutkimuksessa oli 1 768 tekonivelleikkausta, joista 42 infektoitui. Infektoitumattoman tekonivelleikkauksen sairaalakustannukset olivat keskimäärin 7 200 € ja TI:iden 25 100 €. Hoitomenetelmänä kaksivaiheisen revision eli tekonivelen vaihtohoidon hinta oli kolminkertainen tekonivelen säilyttävään hoitoon verrattuna (53 400 € vs. 18 500 €). SI:t ovat yleisiä PPSHP:n alueella sekä terveyskeskusten että OYS:n vuodeosastoilla. SI:iden seurantaohjelma soveltuu infektioseurantaan, mutta se vaatii kohtalaisesti henkilökuntaresurssia. Leikkauksen jälkeinen TI kolminkertaistaa tekonivelleikkauksen sairaalakustannukset.

The diploma thesis deals with problems of healthcare-associated infection in selected nursing activities. The thesis is a theoretical one; it has been processed by means of review and synthesis. In this thesis, four objectives were set, namely to create a comprehensive overview of the problems of healthcare-associated infection in selected nursing activities. Other objectives were to map and analyze clinically relevant sites of transmission and possibilities of prevention of healthcare-associated infection in selected nursing activities and to propose recommendations for the prevention of these infections. The thesis is divided into several chapters that deal with the characteristics of healthcare-associated infection, focusing on the causative agents, formation, spreading and prevention. It also deals with particular nursing activities. The first chosen activity was the care of medical devices intended for repeated use, with a particular focus on the implementation of their disinfection. Secondly, hand sanitation as a basic factor affecting the transmission of healthcare-associated infection activity was selected. This chapter deals primarily with the hygienic hand disinfection, its indications, methods and implementation of disinfection products. It also deals with strategies for improving hand hygiene and glove use in providing nursing care. As the third nursing activity, aseptic techniques, which are an essential part in providing nursing care, were chosen. The thesis describes three specific clinical nursing procedures: aseptic collection of venous blood from a peripheral vein, preparation and administration of a drug into an existing peripheral venous catheter, and uncomplicated (aseptic) wounds dressing. Processing of the thesis was based on a range of relevant sources from the Czech Republic but also from abroad. It also contains various recommendations of national and international institutions. The thesis can serve as a basis for more effective prevention of healthcare-associated infection in certain areas.

Ensuring the bloodstream is one of the most frequently indicated invasive procedures, especially in the pre-hospital emergency and intensive care. All invasive inputs are nowadays an integral part of intensive care. Despite careful daily care, vascular accesses are at risk of complications, which annually effects many patients institutionalized at intensive care units. Catheter sepsis is undoubtedly one of the most serious. Although a significant proportion of these infections are considered highly preventable, they still contribute to high morbidity and mortality among these patients. The aim of this diploma thesis is to analyse nursing care for vascular inputs. One of the partial goals is to evaluate and compare the relevance of the nursing standard of a medical facility with scientific knowledge and current recommendations of professional societies. The research method is a quantitative questionnaire survey focused on nursing staff working in the Department of Anesthesiology and Reanimation. The analysis of the vascular inputs re-dressings was performed in patients hospitalized in the same ward, where the research itself took place. The results of the research survey show a discrepancy between the given procedure in the nursing standard and common practice in the ward. In view of the...

Healthcare-associated infections (HAIs) are used as a measure for federal value-based payment programs. Using data for 2015, the Centers for Disease Control and Prevention (CDC) developed newer risk adjustment models to calculate the standardized infection ratio (SIR) for various infections occurring in hospitals. New national baselines were set to compare performance among medical facilities and states. Despite adjustments for various facility-level factors that contribute to HAI risk, there are ongoing concerns that SIR calculations do not adequately account for non-hospital risk factors that have been linked to clinical outcomes. This explanatory study evaluates state-level data using simple linear regression to determine relationships between the standardized infection ratio (SIR) for methicillin-resistant Staphylococcus aureus (MRSA) bacteremia and several socioeconomic and geographic factors. Bivariate analysis produced significant correlation between SIR and high school education, with states exhibiting lower SIR relative to the percent of adults who completed high school. Higher SIRs were found relative to the percent of state populations subjected to poverty, obesity, and diagnosis of diabetes. Percent of nonprofit hospitals, adults with bachelor’s degrees, and rural residents were not significantly correlated with state measures of MRSA bacteremia. These findings can help guide efforts to reduce HAIs, improve safety of care, and advance population health efforts. The results from this study reinforce the notion that non-hospital factors may have significant effects on the incidence of MRSA bacteremia events occurring in hospitalized patients. Current risk adjustment models that predict MRSA bacteremia events for quality reporting purposes may not adequately account for these risk factors. The present study highlights some ways that hospitals, patients, and policymakers can work together to address social risk factors as a strategy for promoting better and safer care, and healthier communities. This study investigates aspects of the bigger picture of health care quality, performance measurement, and population health. This “feel for the whole” underscores the implications on state performance in infection prevention in the context of socioeconomic and medical vulnerabilities. The study emphasizes the need for greater multidisciplinary collaboration to address community health needs and reduce social and medical disparities.

P. aeruginosa é um importante agente de infecções relacionadas à assistência em saúde. Habitualmente, o estabelecimento de infecções agudas é precedido pela colonização das mucosas dos pacientes. Não se sabe, porém, se os processos infecciosos são causados pelas próprias cepas bacterianas colonizadoras ou por outras com que os pacientes entrem em contato, dotadas ou não de maior potencial de virulência ou de resistência a antimicrobianos que as tornem mais eficientes como agentes infecciosos. Assim, este estudo teve como objetivos i) investigar a existência de potenciais diferenças entre amostras de P. aeruginosa que causaram apenas colonização e aquelas responsáveis por infecção, isoladas de um mesmo paciente, quanto a seus fenótipos de virulência e de não susceptibilidade a antimicrobiamos; ii) pesquisar a existência de associação entre características dos paciente, incluindo o tipo de evolução clínica, com as demais variáveis estudadas. No estudo foram incluídos 21 pacientes que desenvolveram infecção por P. aeruginosa durante sua internação no Centro de Terapia Intensiva do Hospital Universitário Clementino Fraga Filho, entre abril de 2007 e abril de 2008. De cada paciente foram selecionadas duas amostras bacterianas: a primeira isolada durante o episódio de infecção e a amostra colonizadora obtida imediatamente antes da ocorrência da infecção. As amostras selecionadas foram estudadas quanto a i) expressão de três mecanismos de virulência (citotoxicidade, aderência a células epiteliais respiratórias humanas e capacidade de formação de biofilme); ii) presença de genes codificadores das proteínas efetoras do sistema de secreção do tipo 3 (SST3 - exoS, exoT, exoU e exoY); iii) perfil de susceptibilidade a antimicrobianos, iv) perfil de fragmentação do DNA cromossômico por eletroforese em gel de campo pulsado (PFGE). As amostras bacterianas obtidas de infecções agudas foram significativamente mais citotóxicas que aquelas obtidas de colonização. Embora sem diferença estatística, a citotoxicidade das amostras que causaram infecção em pacientes que evoluíram para óbito foi superior à citotoxicidade das amostras de pacientes que sobreviveram. O gene que codifica a toxina ExoU foi detectado em 16 amostras (38%), sendo nove de colonização e sete de infecção. Não houve diferença significativa entre as amostras de colonização e infecção quanto à aderência, produção de biofilme, expressão dos genes do SST3 e não-susceptibilidade às diferentes classes de antimicrobianos. Também não foi encontrada associação entre a não-susceptibilidade à quinolona, ou a outras classes de antimicrobianos, e a presença do gene exoU. As 42 amostras de P. aeruginosa estudadas foram incluídas em 20 genótipos. Em 10 deles foi detectado o gene exoU. Amostras de um mesmo genótipo foram uniformes quanto à expressão dos genes do SST3 e a não-susceptibilidade aos antimicrobianos, mas não quanto às outras variáveis estudadas. Em apenas sete pacientes (33,3%), as amostras de colonização e de infecção pertenciam ao mesmo genótipo. Assim, nesse estudo, o estabelecimento do processo infeccioso resultou não da perda do equilíbrio estabelecido entre os mecanismos de agressão das amostras colonizadoras e os de defesa do hospedeiro e sim da introdução de nova cepa bacteriana no organismo hospedeiro, cepa esta dotada de maior potencial citotóxico. / P. aeruginosa is an important agent of healthcare-associated infections. The establishment of acute infectious episodes is usually preceded by colonization of patient mucosa. However, it remains unknown whether the infectious processes are caused by bacterial strains previously colonizing the patient or by additional strains the patient may come into contact. These new isolates may carry greater virulence potential or antibiotic resistance that makes them more efficient as an infecting agent. Thus, the objetives of the present study were i) to investigate the existence of potential differences between P. aeruginosa isolates obtained from a colonized mucosa and isolates accounting for infectious processes, recovered from the same patient, with respect to virulence phenotypes and non-susceptibility to antimicrobial agents; ii) to investigate the existence of association between patient features, including the type of clinical outcome, with bacterial characteristics. The study included 21 patients who developed P. aeruginosa infection during their stay in the Intensive Care Unit of the University Hospital Clementino Fraga Filho, from April 2007 to April 2008. Two P. aeruginosa isolates were selected from each patient: the first isolate recovered from the infectious episode and the colonizing isolate obtained immediately before the onset of the infection. Features from the isolates investigated included: i) expression of three virulence mechanisms (cytotoxicity, adherence to human respiratory epithelial cells and biofilm formation); ii) presence of the genes encoding type III secretion system effector proteins (TTSS, exoS , exoT , exoU and exoY); iii) antimicrobial susceptibility profile; iv) profile of the bacterial chromossomic DNA fragmentation following analysis by pulsed-field gel electrophoresis (PFGE). The bacterial isolates obtained from acute infections were significantly more cytotoxic than colonizing strains. Moreover, bacteria accounting for infectious episodes in patients who died were more cytotoxic than those recovered from patients who survived, although the differences were not statistically significant. The ExoU toxin encoding gene was detected in 16 (38%) P. aeruginosa isolates: nine colonizing and seven infecting strains. There was no significant difference between colonizing and infecting samples in their adherence, biofilm production, expression of TTSS genes and non- susceptibility to different classes of antimicrobials. There was also no association between non-susceptibility to quinolone, or to any other class of antimicrobial agents, and the presence of the exoU gene. Twenty PFGE genotypes were identified. Isolates from 10 genotypes harboured the exoU gene. Isolates included in the same PFGE genotype exhibited a similar profile of TTSS genes and non-susceptibility to antimicrobials, but not always a similar profile of expression the other variables investigated. In only seven patients (33.3%), the colonizing and infecting isolates belonged to a same genotype. Thus, in this study, the establishment of the infectious process did not result from the loss of the equilibrium established between the aggression mechanisms of colonizing bacteria and host defense but rather from the introduction, in the host organism, of a new bacterial strain, endowed with a greater cytotoxic potential.

P. aeruginosa é um importante agente de infecções relacionadas à assistência em saúde. Habitualmente, o estabelecimento de infecções agudas é precedido pela colonização das mucosas dos pacientes. Não se sabe, porém, se os processos infecciosos são causados pelas próprias cepas bacterianas colonizadoras ou por outras com que os pacientes entrem em contato, dotadas ou não de maior potencial de virulência ou de resistência a antimicrobianos que as tornem mais eficientes como agentes infecciosos. Assim, este estudo teve como objetivos i) investigar a existência de potenciais diferenças entre amostras de P. aeruginosa que causaram apenas colonização e aquelas responsáveis por infecção, isoladas de um mesmo paciente, quanto a seus fenótipos de virulência e de não susceptibilidade a antimicrobiamos; ii) pesquisar a existência de associação entre características dos paciente, incluindo o tipo de evolução clínica, com as demais variáveis estudadas. No estudo foram incluídos 21 pacientes que desenvolveram infecção por P. aeruginosa durante sua internação no Centro de Terapia Intensiva do Hospital Universitário Clementino Fraga Filho, entre abril de 2007 e abril de 2008. De cada paciente foram selecionadas duas amostras bacterianas: a primeira isolada durante o episódio de infecção e a amostra colonizadora obtida imediatamente antes da ocorrência da infecção. As amostras selecionadas foram estudadas quanto a i) expressão de três mecanismos de virulência (citotoxicidade, aderência a células epiteliais respiratórias humanas e capacidade de formação de biofilme); ii) presença de genes codificadores das proteínas efetoras do sistema de secreção do tipo 3 (SST3 - exoS, exoT, exoU e exoY); iii) perfil de susceptibilidade a antimicrobianos, iv) perfil de fragmentação do DNA cromossômico por eletroforese em gel de campo pulsado (PFGE). As amostras bacterianas obtidas de infecções agudas foram significativamente mais citotóxicas que aquelas obtidas de colonização. Embora sem diferença estatística, a citotoxicidade das amostras que causaram infecção em pacientes que evoluíram para óbito foi superior à citotoxicidade das amostras de pacientes que sobreviveram. O gene que codifica a toxina ExoU foi detectado em 16 amostras (38%), sendo nove de colonização e sete de infecção. Não houve diferença significativa entre as amostras de colonização e infecção quanto à aderência, produção de biofilme, expressão dos genes do SST3 e não-susceptibilidade às diferentes classes de antimicrobianos. Também não foi encontrada associação entre a não-susceptibilidade à quinolona, ou a outras classes de antimicrobianos, e a presença do gene exoU. As 42 amostras de P. aeruginosa estudadas foram incluídas em 20 genótipos. Em 10 deles foi detectado o gene exoU. Amostras de um mesmo genótipo foram uniformes quanto à expressão dos genes do SST3 e a não-susceptibilidade aos antimicrobianos, mas não quanto às outras variáveis estudadas. Em apenas sete pacientes (33,3%), as amostras de colonização e de infecção pertenciam ao mesmo genótipo. Assim, nesse estudo, o estabelecimento do processo infeccioso resultou não da perda do equilíbrio estabelecido entre os mecanismos de agressão das amostras colonizadoras e os de defesa do hospedeiro e sim da introdução de nova cepa bacteriana no organismo hospedeiro, cepa esta dotada de maior potencial citotóxico. / P. aeruginosa is an important agent of healthcare-associated infections. The establishment of acute infectious episodes is usually preceded by colonization of patient mucosa. However, it remains unknown whether the infectious processes are caused by bacterial strains previously colonizing the patient or by additional strains the patient may come into contact. These new isolates may carry greater virulence potential or antibiotic resistance that makes them more efficient as an infecting agent. Thus, the objetives of the present study were i) to investigate the existence of potential differences between P. aeruginosa isolates obtained from a colonized mucosa and isolates accounting for infectious processes, recovered from the same patient, with respect to virulence phenotypes and non-susceptibility to antimicrobial agents; ii) to investigate the existence of association between patient features, including the type of clinical outcome, with bacterial characteristics. The study included 21 patients who developed P. aeruginosa infection during their stay in the Intensive Care Unit of the University Hospital Clementino Fraga Filho, from April 2007 to April 2008. Two P. aeruginosa isolates were selected from each patient: the first isolate recovered from the infectious episode and the colonizing isolate obtained immediately before the onset of the infection. Features from the isolates investigated included: i) expression of three virulence mechanisms (cytotoxicity, adherence to human respiratory epithelial cells and biofilm formation); ii) presence of the genes encoding type III secretion system effector proteins (TTSS, exoS , exoT , exoU and exoY); iii) antimicrobial susceptibility profile; iv) profile of the bacterial chromossomic DNA fragmentation following analysis by pulsed-field gel electrophoresis (PFGE). The bacterial isolates obtained from acute infections were significantly more cytotoxic than colonizing strains. Moreover, bacteria accounting for infectious episodes in patients who died were more cytotoxic than those recovered from patients who survived, although the differences were not statistically significant. The ExoU toxin encoding gene was detected in 16 (38%) P. aeruginosa isolates: nine colonizing and seven infecting strains. There was no significant difference between colonizing and infecting samples in their adherence, biofilm production, expression of TTSS genes and non- susceptibility to different classes of antimicrobials. There was also no association between non-susceptibility to quinolone, or to any other class of antimicrobial agents, and the presence of the exoU gene. Twenty PFGE genotypes were identified. Isolates from 10 genotypes harboured the exoU gene. Isolates included in the same PFGE genotype exhibited a similar profile of TTSS genes and non-susceptibility to antimicrobials, but not always a similar profile of expression the other variables investigated. In only seven patients (33.3%), the colonizing and infecting isolates belonged to a same genotype. Thus, in this study, the establishment of the infectious process did not result from the loss of the equilibrium established between the aggression mechanisms of colonizing bacteria and host defense but rather from the introduction, in the host organism, of a new bacterial strain, endowed with a greater cytotoxic potential.

Submitted by JÚLIO HEBER SILVA (julioheber@yahoo.com.br) on 2017-05-12T17:43:06Z No. of bitstreams: 2 Tese - Lillian Kelly de Oliveira Lopes - 2016.pdf: 5335353 bytes, checksum: f43aa281ba6ba16a4f2f6d881c9c2bac (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-05-15T11:47:47Z (GMT) No. of bitstreams: 2 Tese - Lillian Kelly de Oliveira Lopes - 2016.pdf: 5335353 bytes, checksum: f43aa281ba6ba16a4f2f6d881c9c2bac (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-05-15T11:47:47Z (GMT). No. of bitstreams: 2 Tese - Lillian Kelly de Oliveira Lopes - 2016.pdf: 5335353 bytes, checksum: f43aa281ba6ba16a4f2f6d881c9c2bac (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-12-20 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Outro / INTRODUCTION: Depth gauge and flexible drill bite cutter are orthopedic surgical instruments of complex conformation and they are into loanered instrumentation box to different healthcare facilites. The challenge is to ensure that appropriatly cleaning was done to prevent the formation of biofilm in these instruments´models. There are no experimental or clinical studies analyzing the impact of different cleaning protocols on formation of biofilm on surfaces those two surgical instruments. OBJECTIVES: To validate and to evaluate laboratory method to formation of biofilms in stainless steel. To evaluate microbial load and to determine formation of biofilms after laboratory contamination and processing for several cycles in surgical instruments of complex conformation used in surgeries of orthopedic implants. METHOD: Experimental study was developed from November/2014 to March/2016, at the Laboratory of Macquarie University, Sydney (AU). Method´s validation: new forceps halsted hemostatic mosquito were contaminated within contaminant solution containing S. aureus (ATCC 25923) and sterilized in saturated steam. Forceps were divided into three groups according to cleaning: 1) rinsing, 2) manual cleaning and 3) manual cleaning followed by automated cleaning. After 6th, 13th and 20th three forceps from each protocol were analised for microbial load and protein amount (Bicinchoninic Acid Assay) and visual analysis by scanning electronic microscopy. The same method was used on new flexible drill bite cutters and depth gauges, and analised after 10th and 20th reprocessing. RESULTS: Method´s validation: forceps submitted to protocols 1 (positive control of cleaning) and 2 showed coccus in biofilms and forceps sumitted to protocol 3 showed organics residues after 20 reprocessing. No microbial load or residual protein was found. Complex instruments: flexible drill bite cutters submitted to rinsing presented high amount of protein with increase of 1699μg of protein from 10th to 20th reprocessing (P = 0.03). Depth gauge submitted protocol 1 presented high amount of protein, however there was no statistically significant difference from 10th to 20th processing (P = 0.60). All instruments submitted to manual and/or automated cleaning did not present residual protein. It was possible to identify residue and biofilm into lumen of surgical instruments after 20 reprocessing. CONCLUSION: Experimental techniques to formation and evaluation of biofilm in surgical instruments manufactures in stainless steel were validated and 20 inappropriated cleaning processing were enough to buildup biofilm. Biofilm was formed within lumens of flexible drill bite cutter and depth gauge, after 20 processing despite the instruments were submitted to “gold standard” cleaning, also accumulation of protein on flexible drill bite cutters. Depth gauge lumen allowed accumulation of waste using manual cleaning followed by automation cleaning and manual cleaning allowed buildup residue on its external surface. Manual cleaning allowed accumulation residue on deep gauge. It was not possible to recover viable S. aureus in biofilm on instrumental surface, but it indicates that design of instruments evaluated is not safe to processing. It is presumed have risk ratio for aseptic loss of prostheses and infection related to healthcare, since they are instruments of difficult cleaning control and circulate in numerous healthcare facilites using different cleaning protocols. / INTRODUÇÃO: O medidor de profundidade e a fresa flexível são instrumentos cirúrgicos ortopédicos de conformação complexa e integram as caixas cirúrgicas fornecidas por distribuidores aos diferentes serviços de saúde. O desafio é garantir limpeza adequada, impedindo a formação de biofilmes nesses modelos de instrumental. Não existem estudos experimentais ou clínicos analisando o impacto de diferentes protocolos de limpeza na formação de biofilme nas superfícies desses modelos de instrumental. OBJETIVOS: Validar método laboratorial de formação de biofilme e análise em instrumento de aço inoxidável, avaliar a carga microbiana e determinar a formação de biofilme após contaminação laboratorial e processamento por múltiplos ciclos em instrumental cirúrgico de conformação complexa utilizado em cirurgias ortopédicas com implantes. MÉTODO: Estudo experimental realizado de novembro/2014 a março/2016, no laboratório da Universidade Macquarie, Sydney (AU). Validação do método: pinças halsted mosquito novas foram contaminadas em solução contendo S. aureus (ATCC 25923) e esterilizadas em vapor saturado sob pressão. As pinças foram separadas em três grupos segundo a limpeza: 1) enxágue, 2) limpeza manual e 3) limpeza manual seguida de automatizada. Após os 6°, 13° e 20° processamentos, três pinças de cada protocolo foram submetidas à análise de carga microbiana, proteína (Bicinchoninic Acid Assay) e análise visual por microscopia eletrônica de varredura. A mesma metodologia foi seguida no experimento com a fresa flexível e medidor de profundidade novos, sendo as análises realizadas após os 10° e 20° processamentos. RESULTADOS: Validação do método: Pinças submetidas aos protocolos 1 (controle positivo da limpeza) e 2 apresentaram biofilmes, e a pinça submetida ao protocolo 3 apresentou resíduo após 20 processamentos e não foram identificados micro-organismos nem proteína. Experimento com instrumental de conformação complexa: fresa flexível submetida ao protocolo 1 apresentou elevada quantidade de proteína com aumento de 1699µg de proteína do 10° para o 20° processamento (P=0,03). Medidor de profundidade submetido ao mesmo protocolo também apresentou elevada quantidade de proteína, não sendo significante a diferença entre o 10° e o 20° processamento (P=0,60). Todo instrumental submetido à limpeza manual e/ou automatizada não apresentou resíduo de proteína. Foi possível identificar resíduo nos três grupos de limpeza após 20 processamentos. CONCLUSÃO: Técnicas, para formação e avaliação de biofilme em instrumental cirúrgico fabricado em aço inoxidável, foram validadas, e 20 processamentos inadequados formam biofilmes. Houve formação de biofilmes nos lúmens da fresa flexível e no medidor de profundidade após 20 processamentos, mesmo utilizando o padrão ouro da limpeza, bem como acúmulo de proteína na fresa flexível ao longo dos processamentos. O lúmen do medidor de profundidade desmontável permitiu acúmulo de resíduos, mesmo usando limpeza manual seguida de automatizada. Limpeza manual permitiu o acúmulo de resíduos no medidor de profundidade. Não foi possível recuperar S. aureus viáveis no biofilme aderido à superfície de instrumental, porém sinaliza que o design do instrumental avaliado não é seguro para o processamento. Presume-se relação de risco para perda asséptica de próteses e infecção relacionada à assistência à saúde, pois são instrumental de difícil controle da limpeza e circulam em inúmeros serviços de saúdes com diferentes protocolos de limpeza.