Ce travail a pour objet l’étude et le développement d’un capteur électrochimique pour la détection sélective quantitative des analytes biologiques à l’échelle nano. Il se divise en deux parties après une présentation de l’état de l’art sur les maladies nosocomiales et les capteurs électrochimiques. Tout d’abord, nous avons développé un protocole spécifique qui repose sur la fonctionnalisation localisée de la microélectrode de travail par dépôt électrophorétique d'oxyde de graphène réduit/polyéthylènimine (rGO/PEI) pour amplifier le signal de détection. Le microsystème réalisé en salle blanche, a été exploité avec succès pour la détection sélective de la dopamine avec une limite de détection de 50 nM.Ensuite, nous avons utilisé la même plateforme constituée d’électrodes de plus grande taille (mm) pour la réalisation d’un capteur immunologique. Il a démontré son efficacité, de manière spécifique et sélective, pour discriminer la souche sauvage E. Coli UTI89 de UTI89 Δfim (sans opéron), avec une limite de détection de 10 cfu mL−1. En outre, le concept d’utilisation d’une électrode modifiée par rGO/PEI par la modification covalente avec des anticorps pathogènes est général. Il peut être facilement adapté à toute autre espèce pathogène, rendant l’approche générique. Le capteur a donc abouti à des résultats intéressants en milieu aqueux, sérique et urinaire, ce qui est essentiel pour son utilisation potentielle pour le diagnostic clinique des maladies pathogènes. / The purpose of this work concerns the study and development of an electrochemical sensor for both quantitative and selective detection of biological analytes at the nanoscale. It is divided into two parts after a presentation of the state of the art on nosocomial diseases and electrochemical sensors. First, we have developed a specific protocol based on the localized functionalization of the working microelectrode by electrophoretic deposition. The strategy is based on the localizedfunctionalization of the working microelectrode by electrophoretic deposition of reduced graphène oxide / polyethyleneimine (rGO / PEI) to amplify the detection signal. The microsystem built in a clean room has been successfully tested for the selective detection of dopamine with a detection limit of 50 nM. In addition, the microsystem showed good performance in detecting dopamine levels.Then, we have also used the same electrode platform at a larger scale (mm) for the specific and selective detection of the immunological sensor which has demonstrated its effectiveness in distinguishing the UTI89 E. Coli wild-type strain of UTI89 Δfim (without operon), with a detection limit of 10. cfu mL-1. In addition, the concept of rGO / PEI modified electrode by covalent modification with pathogenic antibodies is general and can be easily transposed to any other pathogenic species, making the approach very versatile and generic.The sensor works in aqueous, serum and urinary media, which is essential for its potential use in clinical diagnosis of pathogenic diseases.

Die Mehrzahl der jährlich 400.000 bis 600.000 Krankenhausinfektionen in Deutschland wird von Erregern der sog. ESCAPE-Gruppe (Enterococcus faecium, Staphylococcus aureus, Clostridium difficile, Acinetobacter baumannii, Pseudomonas aeruginosa und verschiedene Enterobacteriaceae, u.a. Klebsiella pneumoniae) verursacht. Besondere Sorge bereitet dabei die Ausbreitung von K. pneumoniae-Stämmen mit enzymvermittelter Resistenz gegenüber Carbapenem-Antibiotika (K. pneumoniae-Carbapenemase, KPC) und die Zunahme von C. difficile-Infektionen (CDI) durch hypervirulente Epidemiestämme (z.B. Ribotyp 027). Die spezifischen Erfahrungen eines prolongierten Ausbruchsgeschehens durch einen KPC-bildenden K. pneumoniae-Stamm (KPC-KP) am Leipziger Universitätsklinikum machen deutlich, dass bei diesem Erregertyp ein hohes Transmissionspotential bei enormer Tenazität (Umweltresistenz) zu berücksichtigen ist, ein Versagen von Standardhygienemaßnahmen in Betracht zu ziehen ist, und Infektionsketten oftmals unklar bleiben. Die Anwendung von Antibiotika ist bei KPC-KP-Infektionen auf einzelne Substanzen (Colistin, Tigecyclin, Gentamicin) beschränkt und vor allem bei immunsupprimierten Patienten (z.B. Lebertransplantierte) mit einem relevanten Risiko des Therapieversagens behaftet. Die Therapie von CDI wird gerade bei Immunsupprimierten durch eine steigende Zahl an Rezidiven erschwert, die teilweise antibiotisch (Vancomycin, Fidaxomicin) nicht beherrschbar sind, so dass alternative Therapieverfahren wie die fäkale Bakterientherapie („Stuhltransplantation“) zur Anwendung kommen. CDI-Rezidive, aber auch eine dauerhafte intestinale Besiedelung mit multiresistenten Enterobakterien wie KPC-KP, scheinen neben wirtsspezifischen Faktoren der Immunantwort durch eine Dysregulation der physiologischen intestinalen Standortflora mit Störung der Kolonisationsresistenz bedingt zu sein. Der Versuch einer Eradikationsbehandlung von Patienten mit persistierender intestinaler Besiedelung durch KPC-KP mittels oraler Applikation der nicht resorbierbaren Antibiotika Colistin und Gentamicin ist mit einem relevanten Risiko der Entstehung von Sekundärresistenzen behaftet. Die Zulassung neuer, besser wirksamer Antibiotika ist für die nächsten Jahre nicht in Sicht, so dass der Infektionsprävention überragende Bedeutung zukommt. Die Erfahrungen der KPC-Ausbruchsbewältigung am Leipziger Universitätsklinikum zeigen, dass nahezu lückenlose Compliance bei der Händedesinfektion, rigoros praktizierte und kontrollierte Barriere- und Isolationsmaßnahmen, Optimierung des Gebrauchs von Breitspektrum-Antibiotika (sog. „Antibiotic Stewardship“) und systematisches mikrobiologisches Erregerscreening dabei unabdingbar sind. Nachhaltige Verbesserungen hinsichtlich der globalen Ausbreitung von multiresistenten Krankenhausbakterien werden sich nur durch grundlegende Umgestaltungen in Umwelt, Landwirtschaft, Tierzucht und Gesundheitswesen mit sparsamer und möglichst gezielter Anwendung von Antibiotika erzielen lassen. Um Risikopopulationen hospitalisierter Patienten vor potentiell lebensbedrohlichen Erregertransmissionen effektiv schützen zu können, sind erweiterte Surveillance und konsequent umgesetzte krankenhaushygienische Maßnahmen erforderlich.

[Truncated abstract] The prinicipal aim of this thesis was to explore the relationship between 3rd generation cephalosporin antibiotics and hospital-acquired Clostridium difficile-associated diarrhoea (CDAD). This antibiotic class has been implicated in the aetiology of CDAD; therefore restriction of these antibiotics via antibiotic policies represents a potential strategy for prevention and control of CDAD. Successful control of CDAD in hospitals translates to improved quality of care for patients, and a reduction of pressure on hospital resources. Therefore, the objectives of this study were to determine whether 3rd generation cephalosporins were related to CDAD, to evaluate the effect of changes to antibiotic policy on the incidence of CDAD, and to determine the impact of CDAD on patient length of stay and hospital costs. The study was conducted in Sir Charles Gairdner Hospital (SCGH), a public teaching hospital located in Perth, the capital city of the state of Western Australia. Evidence for an association between 3rd generation cephalosporins and CDAD was obtained from studies of ecologic- and individual-level data. A time series analysis of the relationship between monthly consumption of 3rd generation cephalosporins and the incidence of CDAD in SCGH was undertaken covering the period 1994 to 2000. The results demonstrated a positive relationship between the use of 3rd generation cephalosporins and CDAD. A matched case-control study that involved 193 adult inpatients diagnosed with CDAD and 386 adult inpatients without CDAD, selected from the period 1996 to 2000, was conducted. Information was collected on exposure to 3rd generation cephalosporin antibiotics during hospitalisation, as well as exposure to other antibiotics and medications, procedures, and comorbidities. Results from conditional logistic regression analyses found CDAD cases were six times more likely to be exposed to 3rd generation cephalosporins during their admission, prior to the onset of diarrhoea, than controls (adjusted odds ratio [OR] = 6.17, 95% confidence interval [CI] = 1.56-24.37). Approximately one third of CDAD in the study population could be attributed to 3rd generation cephalosporins. CDAD cases were also four times more likely to have been exposed to either amoxicillin-clavulanate or ticarcillin-clavulanate (adjusted OR=4.23, 95% CI=1.81-9.93). In October 1998, an antibiotic policy was introduced at SCGH that restricted the use of ceftriaxone, the 3rd generation cephalosporin most commonly used by the hospital. During 1999 and 2000, the incidence of CDAD halved as ceftriaxone consumption fell in response to this policy. The effect of this policy was demonstrated in the time series model; during the post-policy period the relationship between ceftriaxone and CDAD that was evident prior to the policy was cancelled out. From the individual-level data, obtained from the case-control study, a reduction in the prevalence of exposure to 3rd generation cephalosporins from 11% to 1% accounted for a 30% reduction in the incidence of CDAD. Data from the case-control study was also used to analyse the independent contribution of CDAD to length of stay and admission costs using multiple linear regression

La résistance bactérienne aux antibiotiques dans les établissements de santé complique la prise en charge thérapeutique et entraîne une surmortalité des patients infectés. L'objectif de ce travail était d'évaluer différentes approches ayant pour finalité la prévention et la maîtrise des infections nosocomiales. Ce travail s'articule autour de trois questions : (i) Existe-t-il une stratégie de promotion d'un juste usage des antibiotiques à privilégier ? (ii) Comment évaluer la performance en matière d'hygiène des mains ? (iii) Quelles mesures pouvons-nous proposer pour limiter la diffusion de P. aeruginosa ?Nous avons d'abord montré que la diffusion de recommandations sous la forme de guides régionaux ou de messages de pharmacovigilance était suivie d'une modification des prescriptions antibiotiques conformes aux recommandations.Ensuite, nous avons évalué le niveau de performance des établissements de santé en matière d'hygiène des mains en étudiant plus particulièrement l'indicateur de consommation de solution hydro-alcoolique (ICSHA). Nous avons ainsi pu montrer que le nombre minimal d'opportunité d'hygiène des mains servant à son calcul, était sous-estime. Dans une autre étude, nous avons été confrontés aux limites liées à sa construction, ces dernières étant en partie responsable de l'absence de relation observée entre cet indicateur et la prévalence des infections associées aux dispositifs invasifs.Enfin, nous avons montré que la charge en soins et la contamination des réseaux d'eau propre des établissements de santé étaient des facteurs de risque contextuels d'acquisition du bacille pyocyanique. Un autre travail a mis en évidence que les réseaux d'eau usée étaient impliqués dans la diffusion extra-hospitalière de souches résistantes. Une étude est actuellement en cours pour évaluer l'intérêt d'une approche globale associant dépistage et précautions complémentaires chez les patients porteurs de P. aeruginosa.En conclusion, si ce travail confirme l'efficacité de certaines actions de prévention tout en mettant en avant les limites d'autres approches, il ne permet pas de privilégier une stratégie particulière. Il apparaît ainsi nécessaire de mettre en place des stratégies globales et transversales allant au-delà des seuls établissements de santé / Bacterial resistance to antibiotics in health care facilities complicates the therapeutic burden and increased mortality of infected patients. The objective of this work was to evaluate different approaches which aim was to prevent and control hospital-acquired infections. This work focuses on three issues: (i) Is there a strategy already in place to promote the appropriate use of antibiotics? (ii) How can we evaluate performance in terms of hand hygiene? (iii) What measures can we implement to limit the spread of P. aeruginosa?We first showed that the distributions of regional guidelines or drug monitoring alerts were followed by a change in the uptake of antibiotic prescriptions in line with such recommandations.Then we evaluated the performance of health care facilities for hand hygiene, focusing especially on the index of consumption of alcohol-base hand-rub solution. We showed that the number of alcohol-based hand-rub is far higher than that defined by the French Ministry of Health. In another study, we were faced with limitations in its construction, the latter being partly responsible for the lack of a relationship between this indicator and the prevalence of invasive devices associated with infections.Finally, we have shown that the burden of care and the contamination of clean water networks of health facilities were contextual risk factors for acquisition of Pseudomonas aeruginosa. Another study showed that wastewater networks were involved in extra-hospital spread of resistant strains. A study is currently underway to assess the value of a global approach combining screening and additional precautions in patients with P. aeruginosa.In conclusion, this study confirms the effectiveness of some preventive measures while underlining the limitations of other approaches. However it does not promote a particular strategy. Because in terms of BMR, it is necessary to define global and cross-sectorial strategies which go beyond the health care facilities

This thesis deals with the handling of waste and linen. It is divided into a theoretical and practical part.The theoretical part describes the history of Česky Krumlov hospital and is followed by an in-depth look at the history and development of the handling of waste and linen in a medical facility etc. In the practical part of the research qualitative-quantitative methods were used in the form of observation, interview, questionnaire and secondary data analysis. In Česky Krumlov Hospital I observed the ongoing activities and gathered data. The research sample for the interview consisted of 10 respondents. The questionnaires consisted of 21 questions. A secondary analysis of operating rules, guidelines and waste management legislation was used. The main objectives of the research were to compare the system of waste and linen handling in Česky Krumlov hospital with applicable legislation and rules of operation. In so doing, to determine whether there are shortcomings in the separation of waste and the handling of laundry and in which department(s). In addition, to compare the production of waste in amount and time with regards to the use of disposable devices and instruments. Finally, to monitor the way employees separate waste in the hospital and compare the approach to it between smokers and non-smokers.For the purpose of this research five research questions and four hypotheses were put forward: V1: Does the procedure for handling waste meet legislative requirements and the current operating system? V2: Does the system of separation and disposal of waste differ in individual departments and if so why? V3: Can a more effective system for waste management be found? V4: Does the procedure for handling linen meet legislative requirements and the current operating system? V5: Can a more effective way of handling laundry be found? H1: The attitude to the separation of waste is different in smokers and non-smokers. H2: The sorting method depends on education. H3: The sorting method depends on the job position. H4: The sorting method depends on the amount of work experience of employees. Interviews were transcribed using MS Word 2007, and then processed using XMind 6. The hypotheses were statistically analyzed.In addition, information from my observations, interviews and the operating rules and directives for hospital staff helped me to answer the research questions. On the basis of the statistical findings all the hypotheses, with exception to H1, were confirmed. Hypothesis H1 was refuted. During the work all the objectives were met with exception to the fifth objective where problems relating to the quality of services provided by the laundry in České Budějovice Hospital, a.s. were identified. A comparison of the waste management system in Česky Krumlov Hospital a.s. with its operating rules, guidelines and legislation shows that overall the procedures correspond. There were just a few exceptions in the form of unlocked containers for infectious waste and a lack of signatures on labels of some of the staff responsible. During my observation no errors occurred in the separation of waste. The hospital has established a uniform system, whereby the procedures only vary according to the type of waste. The established system of waste separation in Česky Krumlov Hospital a.s. is very good, fully compliant with legislative requirements and is efficient. When comparing the production of general waste over time, the amount is decreasing. However, the amount of hazardous waste being produced is increasing. It was found that the attitude to waste separation does not differ in smokers and non-smokers and that university-educated workers separate less waste at work than others due to the type of job within the organization. Nurses recycle more and more responsibly. The same can be observed for employees with more work experience. I found minor discrepancies in the handling of laundry and the current operating rules.

Os materiais de uso único (MUU) são utilizados há décadas na assistência à saúde. Inicialmente produzidos com a finalidade de prontamente disponibilizar os materiais para uso na assistência a saúde e ao mesmo tempo diminuir a sobrecarga com o trabalho inerente ao reprocessamento dos materiais. Com o passar do tempo, principalmente por questões financeiras, estes passaram a ser reutilizados. A partir de então diversos questionamentos têm sido suscitados a respeito do risco de transmissão de infecção pelo reuso destes artigos em decorrência da dificuldade para de limpeza, desinfecção e esterilização seguras destes. Os materiais utilizados para as cirurgias vídeo-laparoscópicas encontram-se entre os MUU de preço considerável e complexidade importante em relação às dificuldades para limpeza, devido à sua conformação com espaços internos inacessíveis e impossibilidade de desmonte. Isto posto, as pinças grasper, dissector, tesoura, agulha de Veress, sistema de sonda de eletrocirurgia (ou aspirador e irrigador) e sua respectiva haste, instrumentais básicos nos procedimentos de vídeo-cirurgia-laparoscópica, foram o objeto desta investigação com o propósito de avaliar a eficácia da esterilidade destes acessórios de uso único após contaminação artificial desafio e comparar os resultados dos testes de esterilidade dos MUU com os dos materiais equivalentes reprocessáveis – denominados neste estudo como grupo controle. Tratou-se de uma pesquisa experimental, laboratorial e comparativa. O inóculo utilizado para a contaminação artificial dos instrumentais de uso único constituiu-se de suspensão de esporos bacterianos do Bacillus atrophaeus var. niger e de sangue de carneiro desfibrinado esterilizado. Para o grupo controle o microrganismo eleito foi o Geobacillus stearothermophilus acrescido também de sangue de carneiro desfibrinado. Antes de proceder aos experimentos validou-se a metodologia analítica em duas fases: fase 1 - realizada a contagem de U.F.C. em três unidades de cada um dos materiais dos grupos experimental e controle após a contaminação artificial antecedendo a limpeza; fase 2 - realizada a contagem de U.F.C. em três unidades de cada um dos materiais dos grupos experimental e controle após a contaminação artificial e procedimento de limpeza. Desta forma, assegurada a recuperação dos contaminantes em todas as amostras da primeira fase e a diminuição do bioburden inicial após a limpeza dos instrumentais na 2ª fase, deu-se prosseguimento aos experimentos. Os materiais laparoscópicos foram intencionalmente contaminados com o inóculo desafio e submetidos após ao protocolo teste para validação que consistiu da seguinte seqüência: limpeza automatizada em lavadora ultra-sônica com retrofluxo com auxílio do detergente enzimático, limpeza manual complementar com serpilhos e irrigação dos lumens dos instrumentais com água sob pressão. Por fim, foi realizado o enxágüe com água destilada esterilizada, secagem com ar comprimido medicinal, acondicionamento em embalagem de papel grau cirúrgico e filme e submetidos a esterilização em Óxido de Etileno 12/88 (materiais de uso único) e em autoclave (materiais de aço inoxidável). Os materiais esterilizados foram inoculados diretamente nos meios de cultura de caseína soja e incubados por 20 dias quando foi feita a leitura final dos resultados das culturas. Os resultados encontrados foram 100% negativos para a recuperação dos microrganismos contaminantes nos 2 grupos - experimental e controle. Estes achados permitiram concluir que o reprocessamento dos materiais de uso único foi validado em relação à esterilidade permitindo chegar a um protocolo de reprocessamento / Single use devices (SUD) have been used in healthcare for decades. They were initially produced for making them readily available for use in healthcare and at the same time lower the inherent workload for device reprocessing. After time, mainly for financial reasons, they became reused. Since then, several questioning has been raised regarding to the risk of infection transmission by reusing these devices due to their difficulty of safe cleaning, disinfection and sterilization. Devices used for videosurgeries are among the SUD of considerable price and important complexity due to cleaning difficulties, because of their adjustments in inaccessible inner spaces and impossibility of taking them out into pieces. Thus, grasper calipers, dissecting forceps, scissors, Veress insuflation needle, electrosurgery probe system (or suction and irrigation tubes) and its respective dissector cannula, basic devices in videosurgery laparoscopic procedures were all the subject of investigation aimed to evaluate the effectiveness of sterility of these single use products after challenged artificial contamination and comparing the results of sterility tests of SUD to their equivalent reprocessed devices – named in this study as control group. It’s a comparative, laboratory experimental research. The inoculum used for artificial contamination of single use devices was the suspension of Bacillus atrophaeus var. niger bacteria spores and sterilized defibrinated sheep blood. For the control group, the elected strain was Geobacillus stearothermophilus adding also the defibrinated sheep blood. Before carrying out the experiments, the analytical methodology was validated in two phases: phase 1 – counting the colony formation units in three units of each one of the devices from the experimental and control groups after the artificial contamination and cleaning procedures. This way, assured the contaminant recovery in all samples of the 1st phase and the reduction of initial bioburden after device cleaning in the 2nd phase, it was carried on the experiments. The laparoscopic devices were intentionally contaminated with challenged inoculum and submitted after the trial test to validation that was consisted of the following sequence: cleaning in automated narrow-lumen cleaner with enzymatic detergent, complementary manual cleaning with brush and lumen washing of devices with water under pressure. At last, it was made rinsing with sterile distillated water, drying with medical compressed air, packing in surgical paper and film wrapping and submitted to sterilization in ethylene oxide 12/88 (single use devices) and in autoclave (stainless steel devices). The sterilized devices were contaminated directly in the casein soy media and incubated for 20 days when it was made the final reading of media results. The outcomes found were 100% negative for recovering of contamination strains in the 2 groups – the experimental and the control. These findings permitted us to conclude that single use device reprocessing was validated regarding to sterility allowing us to get a reprocessing guideline

O ambiente hospitalar é um local que requer cuidados especiais. No caso de reformas e até mesmo construções um cuidado maior é necessário, já que demolições, lixamentos e outros tipos de intervenções podem gerar aerossóis. Nos aerossóis estão presentes fungos que ao serem inalados por pacientes imunocomprometidos ou transplantados apresentam grande probabilidade de provocar infecções graves, que em muitos casos são letais. Face ao exposto, o presente trabalho tem como objetivo caracterizar a ocorrência de fungos em argamassas de revestimento no ambiente hospitalar. A pesquisa foi realizada em paredes de alvenaria do Hospital de Clínicas, da Universidade Federal do Paraná, de onde foram extraídas amostras da região da superfície, da argamassa e do bloco cerâmico. As amostras foram semeadas em placas de Petri e RODAC® contendo Ágar Sabouraud Dextrose e incubadas a 25oC durante sete dias. Após esse período o crescimento de fungos nas placas foi analisado. A variação da temperatura e umidade foi monitorada por meio de sistema remoto e manual. A argamassa dos ambientes avaliados teve seus valores de resistência à tração e teor de umidade determinados. Das 150 amostras coletadas 39% apresentaram crescimento positivo com a seguinte distribuição de fungos: Aspergillus (presente em 28% das amostras), Absidia (21%), Cladosporium (18%), Rhizopus (10%), Rhodotorulla (8%), Fusarium (6%), Penicillium (3%), A. flavus, demais fungos e outros fungos filamentosos (2%) cada. Nos substratos do interior das paredes pesquisadas foram identificadas três espécies diferentes de Aspergillus: A. flavus, A. fumigatus e A.niger. Todas as espécies de Aspergillus encontradas podem causar aspergilose invasiva e, por isso, oferecem sérios riscos à saúde de pacientes imunocomprometidos. Em uma das paredes avaliadas o teor de umidade encontrado foi igual a 12% e a resistência à tração foi nula. O estudo evidenciou a presença de fungos oportunistas, tais como Aspergillus e Fusarium, tanto nas amostras de argamassa quanto nas de bloco cerâmico retiradas do interior das paredes dos ambientes pesquisados. / The hospital is a place that requires special care. In terms of reforms and even buildings, this site requires even greater care since, demolition, sanding and other types of interventions can generate aerosols. In aerosols, commonly called dust, fungi are present is high the probability of the fungi be aspirated by immunocompromised patients in a hospital. This microorganisms can germinate and cause severe infections and lethal in many cases. The present work has as its object to identify and characterize the occurrence of fungi in mortar coating in the hospital environment. The survey was conducted in masonry walls of the Clinics Hospital, Federal University of Paraná, in which were extracted samples of the surface, of the mortar and of the ceramic block. Samples were plated on Petri plates containing Sabouraud Dextrose Agar, incubated at 25 ° C for seven days, and after this period was analyzed fungal growth. The environments had their temperature and humidity monitored by a remote and manual system during one year. The mortar coating these environments was also investigated with respect to tensile strength and moisture content. Of the samples collected 39% showed growth of colonies with the following distribution of microorganisms: Aspergillus (28% of the samples), Absidia (21%), Cladosporium (18%), Rhizopus (10%), Rhodotorulla (8%), Fusarium (6%), Penicillium (2%), A. flavus, other genera and other filamentous fungi (2%). Within the substrate researched were identified three different species of Aspergillus: A. A.flavus, A.fumigatus and A.niger. All species of Aspergillus found cause invasive aspergillosis and therefore offer a serious risk to immunocompromised patients. In one of the walls evaluated the moisture content was equal 12% and the tensile strength was zero. The study revealed the presence of opportunistic fungi such as Aspergillus, Fusarium, among others, in the mortar and in the ceramic block inside the wall of the environments studied.

O ambiente hospitalar é um local que requer cuidados especiais. No caso de reformas e até mesmo construções um cuidado maior é necessário, já que demolições, lixamentos e outros tipos de intervenções podem gerar aerossóis. Nos aerossóis estão presentes fungos que ao serem inalados por pacientes imunocomprometidos ou transplantados apresentam grande probabilidade de provocar infecções graves, que em muitos casos são letais. Face ao exposto, o presente trabalho tem como objetivo caracterizar a ocorrência de fungos em argamassas de revestimento no ambiente hospitalar. A pesquisa foi realizada em paredes de alvenaria do Hospital de Clínicas, da Universidade Federal do Paraná, de onde foram extraídas amostras da região da superfície, da argamassa e do bloco cerâmico. As amostras foram semeadas em placas de Petri e RODAC® contendo Ágar Sabouraud Dextrose e incubadas a 25oC durante sete dias. Após esse período o crescimento de fungos nas placas foi analisado. A variação da temperatura e umidade foi monitorada por meio de sistema remoto e manual. A argamassa dos ambientes avaliados teve seus valores de resistência à tração e teor de umidade determinados. Das 150 amostras coletadas 39% apresentaram crescimento positivo com a seguinte distribuição de fungos: Aspergillus (presente em 28% das amostras), Absidia (21%), Cladosporium (18%), Rhizopus (10%), Rhodotorulla (8%), Fusarium (6%), Penicillium (3%), A. flavus, demais fungos e outros fungos filamentosos (2%) cada. Nos substratos do interior das paredes pesquisadas foram identificadas três espécies diferentes de Aspergillus: A. flavus, A. fumigatus e A.niger. Todas as espécies de Aspergillus encontradas podem causar aspergilose invasiva e, por isso, oferecem sérios riscos à saúde de pacientes imunocomprometidos. Em uma das paredes avaliadas o teor de umidade encontrado foi igual a 12% e a resistência à tração foi nula. O estudo evidenciou a presença de fungos oportunistas, tais como Aspergillus e Fusarium, tanto nas amostras de argamassa quanto nas de bloco cerâmico retiradas do interior das paredes dos ambientes pesquisados. / The hospital is a place that requires special care. In terms of reforms and even buildings, this site requires even greater care since, demolition, sanding and other types of interventions can generate aerosols. In aerosols, commonly called dust, fungi are present is high the probability of the fungi be aspirated by immunocompromised patients in a hospital. This microorganisms can germinate and cause severe infections and lethal in many cases. The present work has as its object to identify and characterize the occurrence of fungi in mortar coating in the hospital environment. The survey was conducted in masonry walls of the Clinics Hospital, Federal University of Paraná, in which were extracted samples of the surface, of the mortar and of the ceramic block. Samples were plated on Petri plates containing Sabouraud Dextrose Agar, incubated at 25 ° C for seven days, and after this period was analyzed fungal growth. The environments had their temperature and humidity monitored by a remote and manual system during one year. The mortar coating these environments was also investigated with respect to tensile strength and moisture content. Of the samples collected 39% showed growth of colonies with the following distribution of microorganisms: Aspergillus (28% of the samples), Absidia (21%), Cladosporium (18%), Rhizopus (10%), Rhodotorulla (8%), Fusarium (6%), Penicillium (2%), A. flavus, other genera and other filamentous fungi (2%). Within the substrate researched were identified three different species of Aspergillus: A. A.flavus, A.fumigatus and A.niger. All species of Aspergillus found cause invasive aspergillosis and therefore offer a serious risk to immunocompromised patients. In one of the walls evaluated the moisture content was equal 12% and the tensile strength was zero. The study revealed the presence of opportunistic fungi such as Aspergillus, Fusarium, among others, in the mortar and in the ceramic block inside the wall of the environments studied.

The present study sought to establish relationships that exist between the management of solid waste with infectious cases of Infections Related to Health Care (IRHC) in the Adult Intensive Care Unit (ICU-A) of the Emergency Hospital of Sergipe (EHSE), this study aimed to assess the relationship between generators disease vectors found in the referenced industry and infectious waste generated in this health unit. Therefore, it is necessary to: determine how is the handling of infectious solid waste in the ICU-A of a EHSE sector ; identify how the management of infectious waste enables the development of vectors for IRHC , and finally check the preparation of professionals to manage these wastes . Should point out that this is an exploratory research and the field of knowledge was exploited by the inductive method. The literature and empirical basis, about the information necessary for the interpretation and analysis of data is delineated by access to the records of the Committee on Hospital Infection Control (CHIC), which contains the indices of nosocomial infection from 2008 to 2011 and the rules techniques and resolutions codified in chips summaries. Contacts with professionals working in ICU-A and CHIC, support staff and coordination respectively, were also established standardized interviews were conducted, which follow a prescribed script, see Appendix D. The instruments for data collection were used unstructured observation, the field notebook and camera for the purpose of use of the uses of expressions, fragments and images about the current functioning of infectious solid waste management, which are developed based on content analysis. As for the results it was found that among the types of waste generated in the ICU-A EHSE, the sharps are the ones that can be associated with the spread of infectious diseases, these arising from accidents at the time of its disposal, and its occurrence in mostly with housekeeping staff who deal with this type of waste that is commonly and improperly arranged by the clinic staff. However, it became clear that a correct and efficient management of such wastes may prevent scratches and damage in transforming health promotion, and the development of practices for the principles of reverse logistics. / O presente estudo procurou estabelecer que relações existem entre o gerenciamento de resíduos sólidos infectantes com os casos de Infecções Relacionadas à Assistência à Saúde (IRAS) na Unidade de Terapia Intensiva Adulta (UTI-A) do Hospital de Urgência de Sergipe (HUSE), este estudo teve como propósito avaliar a relação entre vetores geradores de doenças encontrados no setor referenciado e os resíduos infectantes gerados nesta unidade de saúde. Para tanto, faz-se necessário: averiguar como é feito o manejo de resíduos sólidos infectantes no setor da UTI-A do HUSE; identificar de que forma o manejo de resíduos infectantes propicia o desenvolvimento de vetores para IRAS; e por fim verificar o preparo dos profissionais para o manejo desses resíduos. Convém pontuar que se trata de uma pesquisa de caráter exploratório e o campo de conhecimento foi explorado pelo método indutivo. A pesquisa bibliográfica e de base empírica, acerca das informações necessárias para interpretação e análise dos dados se delineou mediante o acesso aos registros da Comissão de Controle de Infecção Hospitalar (CCIH), na qual constam os índices de infecção hospitalar de 2008 a 2011 e a normas técnicas e resoluções sistematizadas em fichas resumos. Também foram estabelecidos contatos com os profissionais que trabalham na UTI-A e na CCIH, pessoal de apoio e coordenação respectivamente, foram realizadas entrevistas padronizadas, as quais seguem um roteiro previamente estabelecido, ver apêndice D. Como instrumentos para a coleta de dados foram utilizados a observação não estruturada, o caderno de campo e a máquina fotográfica para efeito de utilização dos usos de expressões, fragmentos e imagens sobre o atual funcionamento da gestão de resíduos sólidos infectantes, sendo estas desenvolvidas com base na análise de conteúdo. Quanto aos resultados foi constatado que dentre os tipos de resíduos gerados na UTI-A do HUSE, os perfurocortantes são os únicos que podem estar associados à propagação de doenças infecciosas, estas, decorrentes de acidentes no momento de sua disposição, sendo sua ocorrência em maior parte com funcionários da limpeza que lidam com este tipo de resíduo e que é comumente disposto inadequadamente pela equipe clínica. No entanto, evidenciou-se que um gerenciamento correto e eficiente desses resíduos pode prevenir riscos e transformar dano em promoção da saúde, como o desenvolvimento de práticas voltadas para os princípios da logística reversa.

Orientador: Maria Luiza Moretti / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-13T02:52:37Z (GMT). No. of bitstreams: 1 Kuboyama_RogerioHakio_D.pdf: 1899697 bytes, checksum: 863826cd141f1e5f89eb90d272c0c330 (MD5) Previous issue date: 2009 / Resumo: Os objetivos do presente estudo, conduzido retrospectivamente, utilizando cepas isoladas de espécimens clínicos obtidos de pacientes internados em dois hospitais brasileiros entre fevereiro de 2001 e junho de 2004 foram descrever: a presença de cepas de Klebsiella pneumoniae produtoras de beta-lactamases de espectro estendido (ESBLs), o melhor método e o substrato preferido para os testes de triagem e de confirmação da produção de ESBLs, a relação epidemiológica das cepas obtidas e analisar os fatores de risco para infecção por cepa produtora de ESBLs. A fim de investigar a relação genética das cepas, foram utilizadas as análises do DNA plasmidial e do DNA cromossômico por eletroforese em campo pulsátil (PFGE). Um total de 89 cepas de K. pneumoniae foram coletadas de diversos sítios anatômicos. Os espécimens clínicos mais comuns dos quais foram isoladas cepas produtoras foram urina (12,4%) e sangue (10,1%). Utilizando os critérios estabelecidos pelo CLSI (testes de triagem), 35 (39,3%) cepas de K. pneumoniae foram consideradas possivelmente produtoras de ESBLs, enquanto que o teste de aproximação de discos (DDAT) revelou distorções características nas zonas de inibição produzidas pela molécula do clavulanato em 96, 62,5, 50, 18,8 e 12,5% das cepas ao redor dos discos contendo aztreonam, cefotaxima, ceftazidima, ceftriaxona e cefpodoxima, respectivamente. Das 89 cepas, 32 (36%) foram consideradas produtoras de ESBLs baseadas no teste confirmatório pelo método Oxoid de discos-combinados. O disco contendo cefotaxima foi capaz de confirmar 100 % dos produtores de ESBLs enquanto que o disco contendo ceftazidima deixou de confirmar 2 cepas produtoras. Dez e 32 diferentes perfis plasmidiais foram observados dentre as cepas de K. pneumoniae produtoras e não produtoras, respectivamente. A PFGE demonstrou melhor poder discriminatório fornecendo 15 e 55 perfis de DNA cromossômico dentre as cepas ESBLs-positivas e ESBLs-negativas, respectivamente. Da análise univariada, as variáveis significativamente associadas com infecção por cepas de K. pneumoniae produtoras de ESBLs foram: uso de cefalosporinas de quarta geração, de lincosamida, de carbapenêmicos, de glicopeptídeos, cirurgia recente, traqueostomia, idade, dias em uso de lincosamida, dias em uso de glicopeptídeos, número total de antibióticos e duração da terapia antimicrobiana. Ao realizar a análise multivariada, utilizando um modelo de regressão logística que incluía as variáveis estatisticamente significantes da análise univariada (P < 0,05), número total de antibióticos permaneceu como única variável independente para infecção por cepa de K. pneumoniae produtora de ESBLs (OR, 1,60; IC95%, 1,194-2,145.; P = 0,0017). Os dados obtidos revelam: uma taxa relativamente alta da produção de ESBLs em cepas de K. pneumoniae obtidas de pacientes das instituições estudadas; a insuficiência da análise plasmidial na elucidação da relação genética dentre as cepas produtoras de ESBLs; aztreonam como melhor substrato indicador da produção presuntiva de ESBLs e cefotaxima como o melhor substrato no teste confirmatório pelo método Oxoid de discoscombinados / Abstract: The objectives of this study conducted restrospectively using strains isolated from clinical specimens obtained from patients hospitalized in two Brazilian hospitals between February 2001 to June 2004 were to describe the presence of extended-spectrum b-lactamase (ESBL)-producing Klebsiella pneumoniae strains, the best method and the preferred substrate for screening and confirming ESBL production and the epidemiological relatedness of ESBL-producing strains and analyse the risk factors for infection due to ESBL-producing K. pneumoniae. To investigate the genetic relatedness of the strains, plasmid analysis and chromosomal DNA analysis by pulsed-field gel electrophoresis were used. A total of 89 K. pneumoniae were collected from diverse body sites. The most commom specimens yielding ESBL-producing strains were urine (12.4%) and blood (10.1%). Using CLSI criteria (ESBL screening breakpoints), 35 K. pneumoniae (39.3%) had presumptive ESBL phenotype, while using the double-disk approximation test (DDAT) characteristic clavulanate-induced distortions of inhibition zones were found in 96, 62.5, 50, 18.8 and 12.5% of the strains around the disks containing aztreonam, cefotaxime, ceftazidime, ceftriaxone and cefpodoxime, respectively. Of 89 isolates, 32 (36%) produced ESBL based on the confirmatory Oxoid combination disk method. The disk containing cefotaxime was able to confirm 100% of the ESBL producers while the disk containing ceftazidime was not able to confirm 2 ESBL-positive strains. Ten and 32 different plasmid profiles were observed among the ESBL-producing K. pneumoniae and non ESBL producers, respectively. Pulsed-field gel electrophoresis showed the best discriminatory power giving 15 and 55 different chromosomal DNA profiles among the ESBL-positive and ESBL-negative K. pneumoniae, respectively. From univariate analysis, variables significantly associated with infection by an ESBL-producing strain of K. pneumoniae included the following: use of 4st-generation cephalosporins, lincosamide, carbapenems, glycopeptides, recent surgery, tracheostomy, age, days in using of lincosamide, days in using of glycopeptides, total number of antibiotics and duration of the antimicrobial therapy. The only variable that remained independent risk factor for acquiring infection due to ESBL-producing K. pneumoniae after multivariable analysis using a logistic regression model, which included the variables associated with acquiring infection by ESBL-producing K. pneumoniae by univariate analysis (P < 0.05), was total number of antibiotics (OR, 1.60; 95%CI, 1.194-2.145; P = 0.0017). In summary, these data indicate that ESBL-producing K. pneumoniae occur at a relatively high incidence at our institutions and the plasmid analysis is not sufficient to identify relationships between ESBL-producing strains of K. pneumoniae. The ESBL screening breakpoints and DDAT with aztreonam appear to be good indicators in presumptive detection of ESBL-producing strains and the cefotaxime used in the Oxoid combination disk method constituted the best substrate in the confirmatory test / Doutorado / Ciencias Basicas / Doutor em Clínica Médica