Detection of mixed microbial biofilms on central venous catheters removed from Intensive care Unit Patients

Cateteres venosos centrais inseridos em pacientes internados em unidade de terapia intensiva foram avaliados por métodos microbiológicos (cultura semi-quantitativa) e microscopia eletrônica de varredura a fim de detectar adesão microbiana e correlacionar com a cultura de sangue. Durante o período de estudo, foram avaliados 59 pacientes com cateter venoso central. A idade dos pacientes, sexo, sítio de inserção e tempo de permanência do cateter foram anotados. O cateter era de poliuretano não tunelizado e de único lúmen. O sangue para cultura foi coletado no momento da remoção do cateter. de 63 pontas de cateteres, 30 (47,6%) foram colonizadas e a infecção encontrada em 5 (23,8%) cateteres. A infecção foi mais prevalente em 26 pacientes (41,3%) com cateteres inseridos em veia subclávia do que nos 3 (3,2%) inseridos em veia jugular. A infecção foi observada com mais freqüência em cateteres com tempo de permanência maior do que sete dias. Os microrganismos isolados incluíram 32 estafilococos coagulase-negativa (29,7%), 61 bactérias Gram-negativas (52,9%), 9 estafilcocos coagulase-positiva (8,3%) e 3 leveduras (2,7%). Como agentes causais de infecções em unidade de terapia intensiva foram isolados E. aerogenes, P. aeruginosa, A. baumannii. Os antimicrobianos com maior atividade in vitro contra as bactérias Gram-negativas foram o imipenem e contra as Gram-positivas vancomicina, cefepime, penicilina, rifampicina e tetraciclina. As análises por microscopia eletrônica de varredura revelaram biofilmes sobre a superfície de todos os cateteres examinados.

Usefulness of catheter tip culture in the diagnosis of neonatal infections

Objective: To determine the number of colony-forming units (CFU) that best correlates with catheter-related infections (CRI) in newborns.Methods: This was a prospective study of semiquantitative cultures of catheter tips obtained from newborns in the neonatal unit at Faculdade de Medicina de Botucatu, state of São Paulo, Brazil. The microorganisms isolated from catheter and peripheral blood cultures were identified and submitted to a drug susceptibility test. The optimal cutoff point was determined by the receiver operating characteristic (ROC) curve.Results: A total of 85 catheters obtained from 63 newborns were studied. Staphylococcus epidermidis was the predominant species in the catheters (75%). Eight of 11 (72.7%) CRI episodes were associated with coagulase-negative staphylococci, six of which were of the S. epidermidis type. ROC curve analysis indicated that the optimal cutoff point for the diagnosis of CRI was 122 CFU.Conclusions: The cutoff point of 122 CFU correlated best with the diagnosis of CRI in newborns.

Caracterização genotípica microrganismos isolados de infecções da corrente sanguína relacionadas a cateteres em recém-nascidos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Análise da segurança oncológica na cirurgia de Wetheim-Meigs via videolaparoscópica comparada à via laparotômica

Pós-graduação em Ginecologia, Obstetrícia e Mastologia - FMB

Assessment of central venous catheter-associated infections using semi-quantitative or quantitative culture methods

Semiquantitative (Maki) and quantitative (Brun- Buisson) culture techniques were employed in the diagnosis of catheter-related bloodstream infections (CRBSI) in patients who have a short-term central venous catheter (inserted for 30 days). The diagnosis of CRBSI was based on the results of semiquantitative and quantitative culture of material from the removed catheters. Catheter tips (118) from 100 patients were evaluated by both methods. Semiquantitative analysis revealed 34 catheters (28.8%) colonized by ≥15 colonyforming units (cfu), while quantitative cultures (34 catheters, 28.8%) showed the growth of ≥103 cfu/mL. Bacteremia was confirmed in four patients by isolating microorganisms of identical species from both catheters and blood samples. Using the semiquantitative culture technique on short-term central venous catheter tips, we have shown that with a cut-off level of ≥15 cfu, the technique had 100.0% sensitivity, specificity of 68.4%, 25.0% positive predictive value (PPV) and 100.0% negative predictive value (NPV), efficiency of 71.4% and a prevalence of 9.5%. The quantitative method, with a cut-off limit of ≥103 cfu/mL, gave identical values: the sensitivity was 100.0%, specificity 68.4%, positive predictive value (PPV) 25.0%, negative predictive value (NPV) 100.0%, efficiency 71.4% and prevalence 9.5%. We concluded that the semiquantitative and quantitative culture methods, evaluated in parallel, for the first time in Brazil, have similar sensitivity and specificity. Keywords: central venous catheter; semi-quantitative culture; quantitative culture; catheter-related bacteremia.

Diagnóstico das infecções relacionadas a cateter em recém-nascidos e epidemiologia molecular dos Staphylococcus spp. isolados

Pós-graduação em Biologia Geral e Aplicada - IBB

Epidemiology of healthcare-associated infections among patients from a hemodialysis unit in southeastern Brazil

Patients submitted to hemodialysis are at a high risk for healthcare-associated infections (HAI). Presently there are scarce data to allow benchmarking of HAI rates in developing countries. Also, most studies focus only on bloodstream infections (BSI) or local access infections (LAI). Our study aimed to provide a wide overview of HAT epidemiology in a hemodialysis unit in southeastern Brazil. We present data from prospective surveillance carried out from March 2010 through May 2012. Rates were compared (mid-p exact test) and temporally analyzed in Shewhart control charts for Poisson distributions. The overall incidence of BSI was 1.12 per 1000 access-days. The rate was higher for patients performing dialysis through central venous catheters (CVC), either temporary (RR = 13.35, 95% CI = 6.68-26.95) or permanent (RR = 2.10,95% CI = 1.09-4.13), as compared to those with arteriovenous fistula. Control charts identified a BSI outbreak caused by Pseudomonas aeruginosa in April 2010. LAI incidence was 3.80 per 1000 access-days. Incidence rates for other HAI (per 1000 patients-day) were as follows: upper respiratory infections, 1.72; pneumonia, 1.35; urinary tract infections, 1.25; skin/soft tissues infections, 0.93. The data point out to the usefulness of applying methods commonly used in hospital-based surveillance for hemodialysis units. (C) 2013 Elsevier Editora Ltda. All rights reserved.

Different outcomes of peritoneal catheter percutaneous placement by nephrologists using a trocar versus the Seldinger technique: the experience of two Brazilian centers

A good catheter implantation technique is important to allow effective peritoneal access function and long-term technique survival. Studies regarding results obtained by nephrologists in comparison with different techniques have been limited. The aim of this study was to investigate the rate of early catheter-related complications and catheter survival in two Brazilian centers, according to two different percutaneous methods of catheter implantation performed by nephrologist team. Adult incident patients recruited from January 2006 to July 2013 having undergone first peritoneal dialysis (PD) catheter implantation were included in the analysis. Mechanical and infectious early complication rates were defined as time to the first event occurring up to 3 months. Four hundred and forty-five consecutive Tenckhoff catheters were implanted by nephrologist team percutaneously after antibiotic prophylaxis in an operating room: trocar was used in 349 (78.4 %) and Seldinger technique (ST) in 99 (21.6 %). The ST was significantly associated with a lower rate of leak (16.3 vs 3 %, p = 0.03) and outflow failure due to tip catheter migration (22.6 vs 10.1 %, p = 0.04), while early infectious complication rates were similar between the two groups (p = 0.59). Long-term catheter survival was higher in Seldinger group (log-rank, p = 0.031). By Cox multivariate analysis, adjusted for age, sex, and diabetes, the ST remained independently associated with better catheter survival [HR 0.681 (0.462-0.910), p = 0.04]. As conclusion, our experience showed better PD outcomes with the ST than trocar method of catheter implantation by nephrologist.

Comparação das técnicas semi-quantitativa e quantitativa nas infecções relacionadas a cateter em neonatos e determinação da resistência à oxacilina nos Staphylococcus spp

Study the semi-quantitative and quantitative technique in the diagnosis of catheter-related infections in newborns and to determine oxacillin resistance in Staphylococcus isolated. It was analyzed 353 catheter tips from 273 newborns in the Neonatal Unit of Hospital FMB. To confirm the diagnosis of infection, were analyzed the clinical data of newborns, the presence of at least one positive blood culture and growth of ≥ 1000 CFU/mL on quantitative culture and/or ³15 UFC on semiquantitative culture, with the same microorganism isolation (species and drug sensitivity) in blood culture and no other focus of infection except the catheter. The disk diffusion technique was used to check similarity of strains and resistance to oxacillin. Of the 353 tips analyzed, 39 were included in this study as the inclusion criteria. The semiquantitative culture was positive in 26 (66.7%) catheters and quantitative culture was positive in 24 (61.5%). Of 273 patients, 19 (6.9%) had a diagnosis of catheter-related bloodstream Infection (CR-BSI). Of the 19 episodes of CR-BSI, S. epidermidis was the predominant etiological agent (84.2%). The resistance to the antibiotic methicillin was found in 14 (73.7%) strains of Staphylococcus. The semiquantitative method was more sensitive (79%) compared with the quantitative method (63%). The use of antibiotics may have influenced the sensitivity of the quantitative method as the microorganisms present in the lumen are exposed to higher concentrations of antibiotics administered via the catheter

Ensaio clínico randomizado do uso do curativo gel de clorexidina para a prevenção da colonização do cateter venoso central em pacientes adultos críticos

Os acessos venosos são indispensáveis para assistência do paciente em situação crítica. O cateter venoso central (CVC) é um acesso que viabiliza a terapêutica dessa clientela, mas o seu uso pode levar à infecções. Estas infecções ocasionam maior permanência hospitalar, elevam os custos totais das instituições e aumentam a morbidade e a mortalidade do paciente. O uso de curativos como cobertura do sítio de saída do CVC é eficaz na prevenção das infecções relacionadas a estes cateteres, em particular, o uso de curativos impregnados com antissépticos como o curativo gel de clorexidina. Este estudo teve como objetivo comparar a efetividade do curativo gel de clorexidina com a do filme transparente de poliuretano na prevenção da colonização do cateter venoso central em pacientes adultos críticos. Trata-se de estudo experimental, do tipo ensaio clínico randomizado, com tratamentos em paralelo, prospectivo e monocêntrico, realizado de acordo com as recomendações do Consolidated Standards of Reporting Trials (CONSORT). O estudo foi realizado na Unidade de Terapia Intensiva e na Unidade Coronariana de um hospital de ensino do interior do Estado de São Paulo. Participaram do estudo 102 indivíduos hospitalizados nestes locais, divididos aleatoriamente em dois grupos: grupo intervenção, no qual o tipo de cobertura utilizada foi o curativo de gel de clorexidina e grupo controle, que utilizou como cobertura o filme transparente de poliuretano. O desfecho primário mensurado foi a colonização do cateter e os desfechos secundários foram a infecção clínica do sítio de saída, a infecção microbiológica do sítio de saída e a infecção da corrente sanguínea relacionada ao cateter. Para a coleta de dados foi elaborado um instrumento, e este validado quanto ao seu conteúdo e forma por 13 enfermeiros pertencentes aos locais do estudo. Estes profissionais foram treinados para a realização dos curativos e coleta das pontas dos cateteres centrais, swabs dos sítios de saída e hemoculturas. Análises descritivas foram usadas para todas as variáveis do estudo. O teste Exato de Fisher foi utilizado para comparar as proporções de cada desfecho nos grupos de intervenção e controle, e a regressão logística para explorar se a colonização no CVC poderia ser associada com o tempo de uso do cateter e com o Acute Physiology and Chronic Health Evaluation II (APACHE II) dos pacientes do estudo. De acordo com os resultados não houve diferença estatisticamente significante entre a colonização nos dois grupos (p valor = 1.00), para a infecção microbiológica do sítio de saída (p valor = 0.08), para a infecção clínica do sítio de saída (p valor = 0.77) e para as infecções da corrente sanguínea relacionadas ao cateter (p valor = 1,00). Conclui-se que o presente estudo pode contribuir para que as unidades de saúde tenham subsídios para realizar a escolha do tipo de curativo baseado em suas necessidades institucionais e no desenvolvimento de protocolos relacionados à medidas de inserção e manutenção do cateter, bem como medidas educativas permanentes

Randomized, controlled trial of topical exit-site application of honey (Medihoney) versus Mupirocin for the prevention of catheter-associated infections in hemodialysis patients

The clinical usefulness of hemodialysis catheters is limited by increased infectious morbidity and mortality. Topical antiseptic agents, such as mupirocin, are effective at reducing this risk but have been reported to select for antibiotic-resistant strains. The aim of the present study was to determine the efficacy and the safety of exit-site application of a standardized antibacterial honey versus mupirocin in preventing catheter-associated infections. A randomized, controlled trial was performed comparing the effect of thrice-weekly exit-site application of Medihoney versus mupirocin on infection rates in patients who were receiving hemodialysis via tunneled, cuffed central venous catheters. A total of 101 patients were enrolled. The incidences of catheter-associated bacteremias in honey-treated (n = 51) and mupirocin-treated (n = 50) patients were comparable (0.97 versus 0.85 episodes per 1000 catheter-days, respectively; NS). On Cox proportional hazards model analysis, the use of honey was not significantly associated with bacteremia-free survival (unadjusted hazard ratio, 0.94; 95% confidence interval, 0.27 to 3.24; P = 0.92). No exit-site infections occurred. During the study period, 2% of staphylococcal isolates within the hospital were mupirocin resistant. Thrice-weekly application of standardized antibacterial honey to hemodialysis catheter exit sites was safe, cheap, and effective and resulted in a comparable rate of catheter-associated infection to that obtained with mupirocin (although the study was not adequately powered to assess therapeutic equivalence). The effectiveness of honey against antibiotic-resistant microorganisms and its low likelihood of selecting for further resistant strains suggest that this agent may represent a satisfactory alternative means of chemoprophylaxis in patients with central venous catheters.

Induction of inflammatory cytokines and nitric oxide in J774.2 cells and murine macrophages by lipoteichoic acid and related cell wall antigens from Staphylococcus epidermidis

Staphylococcus epidermidis causes infections associated with medical devices including central venous catheters, orthopaedic prosthetic joints and artificial heart valves. This coagulase-negative Staphylococcus produces a conventional cellular lipoteichoic acid (LTA) and also releases a short-glycerophosphate-chain-length form of LTA (previously termed lipid S) into the medium during growth. The relative pro-inflammatory activities of cellular and short-chain-length exocellular LTA were investigated in comparison with peptidoglycan and wall teichoic acid from S. epidermidis and LPS from Escherichia coli O111. The ability of these components to stimulate the production of proinflammatory cytokines [interleukin (IL)-1β, IL-6 and tumour necrosis factor (TNF)-α] and nitric oxide was investigated in a murine macrophage-like cell line (J774.2), and in peritoneal and splenic macrophages. On a weight-for-weight basis the short-chain-length exocellular LTA was the most active of the S. epidermidis products, stimulating significant amounts of each of the inflammatory cytokines and nitric oxide, although it was approximately 100-fold less active than LPS from E. coli. By comparison the full-chain-length cellular LTA and peptidoglycan were less active and the wall teichoic acid had no activity. As an exocellular product potentially released from S. epidermidis biofilms, the short-chain-length exocellular LTA may act as the prime mediator of the host inflammatory response to device-related infection by this organism and act as the Gram-positive equivalent of LPS in Gram-negative sepsis. The understanding of the role of short-chain-length exocellular LTA in Gram-positive sepsis may lead to improved treatment strategies. © 2005 SGM.

Serodiagnosis of staphylococcal sepsis

The coagulase-negative staphylococci are the most frequent cause of sepsis associated with indwelling intravascular catheters. Current microbiological investigations to support the diagnosis of catheter-related sepsis (CRS) include the culture of blood and catheter tips, however positive results may reflect specimen contamination, or colonisation of the catheter rather than true sepsis. Previous serological approaches to assist in the diagnosis of CRS based on cellular staphylococcal antigens have been of limited value. In this current study, the serodiagnostic potential of an exocellular antigen produced by 7 strains of coagulase-negative staphylococci cultured in brain heart infusion broth was investigated. Antigenic material isolated by gel permeation from liquid culture was characterised by immunological techniques and chemical analysis. Characterisation of the exocellular antigen revealed a novel glycerophosphoglycolipid, termed lipid S. which shared antigenic determinants with lipoteichoic acid, but differed by comprising a glycerophosphate chain length of only 6 units. In addition, lipid S was immunologically distinct from diphosphatidyl glycerol, a constituent cell membrane phospho lipid. An indirect enzyme linked immunosorbent assay (ELISA) based on lipid S was subsequently developed and used to determine serum antibody levels (IgM and IgG) in 67 patients with CRS due to staphylococci, and 67 patients with a central venous catheter (CVC) in situ who exhibited no evidence of sepsis. The sensitivity and specificity of the lipid S IgG ELISA was 75% and 90% respectively whilst the IgM assay had sensitivity and specificity of 52% and 85%. The addition of GullSORereagent to the EL1SA procedure to remove competing serum IgG and rheumatoid factor did not significantly improve the performance of the IgM assay. The serological response in serial serum samples of 13 patients with CRS due to staphylococci was investigated. Elevated levels of antibody were detected at an early stage of infection, prior to the isolation of microorganisms by standard culture methods, and before the clinical presentation of sepsis in 3 patients. The lipid S ELISA was later optimised and a rapid 4-hour assay developed for the serodiagnosis of CRS. Serum IgG levels were determined in 40 patients with CRS due to staphylococci and 40 patients with a CVC in situ who exhibited no evidence of sepsis. The sensitivity and specificity of the rapid IgG assay was 70% and 100% respectively. Elevated serum antibody levels in patients with endocarditis, prosthetic joint infection and pyogenic spondylodiscitis due to Gram-positive cocci were also detected with the lipid S ELISA suggesting that the assay may facilitate the diagnosis of these infections. Unexpected increased levels of anti-lipid S IgG in 31% of control patients with sciatica suggested a possible microbial aetiology of this condition. Further investigation of some of these patients by culture of microdiscectomy tissue removed at operation, revealed the presence of low-virulent microorganisms in 37% of patients of which Propionibacterium aeries accounted for 85% of the positive culture isolates. The results suggested a previously unrecognised association between P. acnes and sciatica, which may have implications for the future management of the condition.

A prospective clinical trial to evaluate the microbial barrier of a needleless connector

Needleless connectors are being increasingly used for direct access to intravascular catheters. However, the potential for microbial contamination of these devices and subsequent infection risk is still widely debated. In this study the microbial contamination rate associated with three-way stopcock luers with standard caps attached was compared to those with Y-type extension set luers with Clearlink® needleless connectors attached. Fifty patients undergoing cardiothoracic surgery who required a central venous catheter (CVC) as part of their peri- and postoperative management were studied for microbial contamination of CVC luers following 72 hrs in situ. Each patient's CVC was randomly designated to have either the three-way stopcocks with caps (control patients) or Clearlink® Y-type extension sets (test patients). Prior to, and following each manipulation of the three-way stopcock luers or Clearlink® devices, a 70% (v/v) isopropyl alcohol swab was used for disinfection of the connections. The microbial contamination of 393 luers, 200 with standard caps and 193 with Clearlink® attached, was determined. The internal surfaces of 20 of 200 (10%) three-way stopcock luers with standard caps were contaminated with micro-organisms whereas only one of 193 (0.5%) luers with Clearlink® attached was contaminated (P < 0.0001). These results demonstrate that the use of the Clearlink® device with a dedicated disinfection regimen reduces the internal microbial contamination rate of CVC luers compared with standard caps. The use of such needle-free devices may therefore reduce the intraluminal risk of catheter-related bloodstream infection and thereby supplement current preventive guidelines. © 2006 The Hospital Infection Society.

Using the collaborative evidence based practice model : a systematic review and uptake of chlorhexidine-impregnated sponge dressings on central venous access devices in a tertiary cancer centre

Background: Greater research utilisation in cancer nursing practice is needed, in order to provide well-informed and effective nursing care to people affected by cancer. This paper aims to report on the implementation of evidence-based practice in a tertiary cancer centre. Methods: Using a case report design, this paper reports on the use of the Collaborative Model for Evidence Based Practice (CMEBP) in an Australian tertiary cancer centre. The clinical case is the uptake of routine application of chlorhexidine-impregnated sponge dressings for preventing centrally inserted catheter-related bloodstream infections. In this case report, a number of processes that resulted in a service-wide practice change are described. Results: This model was considered a feasible method for successful research utilisation. In this case report, chlorhexidine-impregnated sponge dressings were proposed and implemented in the tertiary cancer centre with an aim of reducing the incidence of centrally inserted catheter-related bloodstream infections and potentially improving patient health outcomes. Conclusion: The CMEBP is feasible and effective for implementing clinical evidence into cancer nursing practice. Cancer nurses and health administrators need to ensure a supportive infrastructure and environment for clinical inquiry and research utilisation exists, in order to enable successful implementation of evidence-based practice in their cancer centres.