C-reactive protein and albumin kinetics before community-acquired bloodstream infections - a Danish population-based cohort study.

Early changes in biomarker levels probably occur before bloodstream infection (BSI) is diagnosed. However, this issue has not been fully addressed. We aimed at evaluating the kinetics of C-reactive protein (CRP) and plasma albumin (PA) in the 30 days before community-acquired (CA) BSI diagnosis. From a population-based BSI database we identified 658 patients with at least one measurement of CRP or PA from day -30 (D-30) through day -1 (D-1) before the day of CA-BSI (D0) and a measurement of the same biomarker at D0 or D1. Amongst these, 502 had both CRP and PA measurements which fitted these criteria. CRP and PA concentrations began to change inversely some days before CA-BSI diagnosis, CRP increasing by day -3.1 and PA decreasing by day -1.3. From D-30 to D-4, CRP kinetics (expressed as slopes - rate of concentration change per day) was -1.5 mg/l/day. From D-3 to D1, the CRP slope increased to 36.3 mg/l/day. For albumin, the slope between D-30 to D-2 was 0.1 g/l/day and changed to -1.8 g/l/day between D-1 and D1. We showed that biomarker levels begin to change some days before the CA-BSI diagnosis, CRP 3.1 days and PA 1.3 days before.

A novel in vitro model for haematogenous spreading of S. aureus device biofilms demonstrating clumping dispersal as an advantageous dissemination mechanism.

Staphylococcus aureus is able to disseminate from vascular device biofilms to the blood and organs, resulting in life-threatening infections such as endocarditis. The mechanisms behind spreading are largely unknown, especially how the bacterium escapes immune effectors and antibiotics in the process. Using an in vitro catheter infection model, we studied S. aureus biofilm growth, late-stage dispersal, and reattachment to downstream endothelial cell layers. The ability of the released biofilm material to resist host response and disseminate in vivo was furthermore studied in whole blood and phagocyte survival assays and in a short-term murine infection model. We found that S. aureus biofilms formed in flow of human plasma release biofilm thromboemboli with embedded bacteria and bacteria-secreted polysaccharides. The emboli disseminate as antibiotic and immune resistant vehicles that hold the ability to adhere to and initiate colonisation of endothelial cell layers under flow. In vivo experiments showed that the released biofilm material reached the heart similarly as ordinary broth-grown bacteria but also that clumps to some extend were trapped in the lungs. The clumping dispersal of S. aureus from in vivo-like vascular biofilms and their specific properties demonstrated here help explain the pathophysiology associated with S. aureus bloodstream infections.

Hypoalbuminaemia as a marker of trans-capillary leakage in community-acquired bacteraemia patients.

Community-acquired bacteraemia patients (n = 2472), Denmark, 2000-2008. Albumin, C-reactive protein (CRP) and haemoglobin (Hb) measured 2000-2010. We assessed daily mean levels of albumin, CRP and Hb from 30 days before to 30 days after bacteraemia and correlations between albumin vs. CRP and albumin vs. Hb. In linear regression models, we evaluated the contribution of CRP, Hb, chronic and acute variables to the albumin level variations. The mean albumin level (33.6 g/l) was steady before day 1, declined to 29.3 g/l on day 1 with little increase afterward. The mean CRP increased from day -5, peaked on day 1 and declined thereafter. The mean Hb level was fairly constant during days -30/30. Albumin was inversely (R range, - 0.18/-0.47, P < 10-4) correlated with the CRP level and positively (R = 0.17-0.46, P < 10-4) correlated with the HB level. In most models, CRP was the first variable that contributed to the albumin variations, 34-70% of the full model. The sudden decrease of albumin levels, without sudden fluctuations of CRP or Hb, indicated that hypoalbuminaemia was a marker of trans-capillary leakage.

Bacteremia with Streptococcus pneumoniae: sepsis and other risk factors for 30-day mortality--a hospital-based cohort study.

We conducted a hospital-based cohort study among adult patients with first-time Streptococcus pneumoniae bacteremia (SPB) from 2000 through 2008. Patients were identified in a population-based bacteremia database and followed up for mortality through the Danish Civil Registration System (CRS). The aim of the study was to determine the focal diagnosis of SPB, the severity of sepsis at presentation, demographics and comorbidity characteristics of the patients, and to determine the 30-day mortality rate and factors related to mortality. We identified 481 patients, of which 238 were males. The mean age of the patients was 65 years. The focal diagnosis of the SPB was pneumonia in 381 (79 %) patients, followed in frequency by meningitis in 33 (7 %) patients. Of the 481 patients, 390 (81 %) had community-acquired SPB. Of these, 23 (6 %) did not have sepsis, 132 (34 %) had sepsis, 224 (57 %) had severe sepsis, and 11 (3 %) were in septic shock. Overall, the 30-day mortality was 16 %. Mortality increased with the severity of sepsis. There was no association between the focal diagnosis of SPB or the number of diagnoses and mortality. Nosocomial infection, male sex, increasing age, and increasing comorbidity were all associated with an increased 30-day mortality rate.

Core genome multi-locus sequence typing as an essential tool in a high-cost livestock-associated meticillin-resistant Staphylococcus aureus CC398 hospital outbreak.

BACKGROUND: Livestock-associated meticillin-resistant Staphylococcus aureus (LA-MRSA) clonal complex (CC) 398 may be transmitted and cause morbidity and mortality in hospitals. The economic cost of stopping hospital transmission of LA-MRSA CC398 is poorly described. Early detection of transmission may limit the extent of the intervention. AIM: To evaluate core genome multi-locus sequence typing (cgMLST) for detecting transmission chains and to estimate the costs for interventions to prevent further spread after discovery of hospital transmission of LA-MRSA CC398. METHODS: Five patients were involved in two episodes of transmission of LA-MRSA CC398 in a hospital. Standard interventions including MRSA screening of patients and healthcare workers were initiated. Whole genome sequences of the five isolates and 17 epidemiologically unrelated MRSA CC398 isolates from other hospitalized patients were analysed by single nucleotide polymorphism (SNP) comparisons and cgMLST. The economic costs of constraining transmission were calculated from relevant sources. FINDINGS: The five isolates suspected to be involved in hospital transmission clustered with ≤2 SNPs in the draft genome sequences with some distance to other isolates. cgMLST allocated the five isolates to the same type, which was different from all but two of the sporadic isolates. Furthermore, cgMLST separated the five transmission isolates from all other isolates. The economic costs of the outbreak interventions exceeded €11,000 per patient. CONCLUSION: LA-MRSA CC398 is transmittable in hospitals, and intervention against transmission may reach considerable costs. cgMLST is useful in surveillance of hospital transmission of LA-MRSA.

Compliance with and short-term adverse events from clarithromycin versus placebo in patients with stable coronary heart disease: the CLARICOR trial.

BACKGROUND: The randomized placebo-controlled double-blind CLARICOR trial investigated the influence of clarithromycin versus placebo on cardiovascular events and mortality in patients with chronic coronary artery disease (ClinicalTrials.gov NCT 00121550). The trial randomized 2172 patients to 500 mg of clarithromycin daily versus 2200 patients to matching placebo for 14 days. This paper presents protocol-specified analysis of the patient-reported information regarding their compliance and non-serious adverse events during the 14 days of treatment as well as serious adverse events (mortality and hospitalizations) during the first 30 days after randomization. METHODS: Randomized clinical trial focusing on patient-reported information regarding their compliance and adverse events. RESULTS: Of the randomized patients, 99% reported information regarding their compliance and adverse events. A 100% tablet intake was reported by 90% of the clarithromycin group and by 93.7% of the placebo group. Of the clarithromycin patients, 39.5% reported at least one non-serious adverse event versus 25.1% of the placebo patients (P < 0.001). Gastrointestinal adverse reactions were reported 950 times by 697 patients (32.3%) in the clarithromycin group and 485 times by 390 patients (17.9%) in the placebo group (P < 0.001). No significant differences were seen in other non-serious or serious adverse events during the first month after inclusion. Short-term non-serious adverse events did not explain the previously reported long-term significantly increased mortality associated with clarithromycin. CONCLUSIONS: Gastrointestinal adverse reactions are common during clarithromycin administration, but at least half are also seen with a placebo.

No evidence for presence of bacteria in modic type I changes.

BACKGROUND: Recent studies suggest an association between sciatica and Propionibacterium acnes. "Modic type I changes" in the vertebrae are closely associated with sciatica and lower back pain, and recent studies have questioned the ability of conventional magnetic resonance imaging (MRI) to differentiate between degenerative Modic type I changes and vertebral abnormalities caused by infection. PURPOSE: To test whether bacteria could be cultured from biopsies of Modic type I changes. MATERIAL AND METHODS: Twenty-four consecutive patients with Modic type I changes in lumbar vertebrae had a biopsy taken from the affected vertebra by a strict aseptic procedure. The biopsy was split into two specimens, which were inoculated into thioglycolate agar tubes in the surgical theatre and transported to the microbiology laboratory. In the laboratory, one specimen was streaked onto plates and analyzed for anaerobic and aerobic culture. The other tube was left unopened and incubated directly. Plates and tubes were incubated for 2 weeks and observed for visible growth. RESULTS: None of the biopsies yielded growth of anaerobic bacteria. In one patient, both biopsies yielded growth of Staphylococcus epidermidis, and in another patient coagulase-negative staphylococci were isolated from one biopsy. Both patients received oral antibiotics without convincing effect on symptoms. CONCLUSION: Our results showed no evidence of bacteria in vertebrae with Modic type I changes. The isolation of staphylococci from two patients probably represented contamination.

Shiga toxin-producing Escherichia coli: incidence and clinical features in a setting with complete screening of patients with suspected infective diarrhoea.

OBJECTIVES: Shiga toxin-producing Escherichia coli (STEC) causes diarrhoeal disease, bloody diarrhoea, and haemolytic uraemic syndrome. The aim of this study was to describe the incidence of STEC and the clinical features of STEC patients from a well-defined Danish population in which all fecal samples of patients with suspected infective gastroenteritis were analysed for STEC. METHODS: In this population-based cohort study, all stool samples referred to two clinical microbiology laboratories were screened for STEC by culture and/or PCR. Epidemiological (n=170) and clinical (n=209) characteristics were analysed using data from local and national registries. RESULTS: Overall, 75,132 samples from 30,073 patients were screened resulting in 217 unique STEC-isolates. The epidemiological analysis showed an incidence of 10.1 cases per 100,000 person-years, which was more than twofold higher than the incidence in the rest of Denmark (3.4 cases per 100,000 person-years, p <0.001). Three groups were associated with a higher incidence: age <5 years (n=28, p <0.001), age ≥65 years (n=38, p 0.045), and foreign ethnicity (n=27, p 0.003). In the clinical analysis, patients with STEC harbouring only the Shiga toxin 1 gene (stx1-only isolates) showed a lower frequency of acute (n=11, p <0.05) and bloody diarrhoea (n=5, p <0.05) and a higher frequency of gastrointestinal symptoms for ≥3 months (n=8, p <0.05) than the other STEC patients. CONCLUSIONS: We report a more than twofold higher incidence in the project area compared with the rest of Denmark, indicating that patients remain undiagnosed when selective STEC screening is used. We found an association between patients with stx1-only isolates and long-term gastrointestinal symptoms.

Detection and identification of Anaplasma phagocytophilum, Borrelia burgdorferi, and Rickettsia helvetica in Danish Ixodes ricinus ticks.

Borreliosis is an endemic infection in Denmark. Recent serosurveys have indicated that human anaplasmosis may be equally common. The aim of this study was to look for Anaplasma phagocytophilum and related pathogens in Ixodes ricinus ticks and estimate their prevalence, compared to Borrelia, using PCR. Ticks were collected from three locations in Denmark: Jutland, Funen, and Bornholm. Ticks from Jutland and Funen were analysed individually, ticks from Bornholm were analysed in pools of 20. A. phagocytophilum was found in ticks from all areas. A. phagocytophilum was found in 23.6% of ticks from Jutland and Funen, while 11% were positive for Borrelia burgdorferi. The Borrelia genotype B. afzelii was most prevalent, followed by B. valaisiana, B. burgdorferi s.s. and B. garinii.A. phagocytophilum was found in 14.5% of nymphs and 40.5% of adult ticks, while Borrelia was found in 13% of nymphs and 8% of adult ticks. The difference in prevalence between Anaplasma and Borrelia in adult ticks supports the idea that their maintenance cycles in nature may be different. Ticks were also infected with Rickettsia helvetica. Our study indicates that A. phagocytophilum prevalence in ticks in Denmark is as high as Borrelia prevalence and that human anaplasmosis may be unrecognized.

Development of cytomegalovirus (CMV) disease may be predicted in HIV-infected patients by CMV polymerase chain reaction and the antigenemia test.

OBJECTIVE: Cytomegalovirus (CMV) is a frequent opportunistic viral pathogen in patients with AIDS leading to retinitis and other serious manifestations. CMV disease may be successfully treated. Prophylactic antiviral therapy has been shown to reduce the risk of CMV disease if initiated early. We evaluated PCR and the antigenemia tests as methods for early detection of CMV disease. METHODS: Two-hundred HIV-seropositive subjects with CD4 T-cell counts below 100 x 10(6)/l were monitored with CMV polymerase chain reaction (PCR), the antigenemia test, blood cultures and CMV immunoglobulin (Ig) G and IgM titres every second month for 1 year. RESULTS: Thirty-eight patients (19%) developed CMV disease. The PCR test detected CMV DNA a median of 46 days before onset of disease. This was earlier than the median of 34 for the antigenemia test and a median of 1 day for CMV blood cultures. Univariate analysis showed that the CMV PCR, the antigenemia test and blood cultures all had significant predictive values for subsequent development of CMV disease with odds ratios (OR) of 30, 22 and 20. CMV serology had no predictive value. Multivariate analysis showed that the PCR method was superior to the other tests; OR: CMV PCR 10.0, antigenemia test 4.4 and CMV cultures 4.3. No clinical parameters had any significant predictive value in the stepwise multivariate model. CONCLUSIONS: The CMV PCR and the CMV antigenemia tests are both sensitive methods that may predict development of CMV disease up to several months prior to clinical disease. These methods make it possible to select patients at high risk for CMV disease and suitable for prophylactic therapy against CMV.

Comparison of methenamine silver nitrate and Giemsa stain for detection of Pneumocystis carinii in bronchoalveolar lavage specimens from HIV infected patients.

Seventy-seven bronchoalveolar lavages from human immunosuppressive virus infected patients with pulmonary symptoms were examined routinely for the presence of Pneumocystis carinii, using Giemsa stain to detect trophozoites and methenamine silver nitrate to detect cysts. Thirty-seven samples were found positive with both methods, but a further 10 were diagnosed with Giemsa indicating that the trophozoite stain is more sensitive. As Giemsa stain is simple, quick, cheap and familiar to most microbiological laboratories it should be used for screening of samples to be examined for Pneumocystis carinii. In cases where trophozoites are not detected by Giemsa a supplementary cyst stain may be performed in order to rule out the presumably few cases where cysts are present as the only evidence of Pneumocystis carinii infection.

Ribotyping of selected isolates of Enterobacter cloacae and clinical data related to biotype, phage type, O-serotype, and ribotype.

In order to evaluate if ribotyping of selected isolates of Enterobacter cloacae could further elucidate the epidemiology, we performed ribotyping of 109 isolates indistinguishable by bio-, phage-, and O-serotyping, with inconclusive typing results, or from patients with more than one isolate. Ribotyping provided additional information, and some cases of cross-infection or common source of infection were revealed. Under the supposition that isolates sharing the same ribotype were of the same origin, problems arose with respect to bio-, O-sero, as well as phage typing; in particular a remarkable number of isolates showed differences in phage type. In order to identify possible virulence characteristics of certain types, clinical data were related to bio-, phage-, O-sero-, and ribotype. Biotype 66 was significantly more frequent among blood culture isolates (P = 0.001), but this might have reflected the presence of a certain strain in the environment of the intensive care unit, where patients were more likely to develop bacteraemia; serotype 04 was significantly more frequent among isolates from the urinary tract (P = 0.02), and serotype 013 was more frequent among women (P = 0.05). One ribotype was found only among community-acquired isolates, which might suggest that it is a frequent but less virulent strain.

Resistance characteristics of blood culture isolates of Enterobacter cloacae with special reference to beta-lactamases and relation to preceding antimicrobial therapy.

Resistance characteristics of 53 blood culture isolates of E. cloacae were examined and correlated with antimicrobial treatment preceding bacteraemia. Resistance patterns of 22 antimicrobial agents, presence of resistant mutants, and inducibility of beta-lactamase were investigated; furthermore, population analysis and investigation of beta-lactamase production of selected isolates were performed. Thirty-two isolates (60%) were resistant to cephalothin and/or cefoxitin and/or ampicillin, and 14 isolates (26%) had further resistance characteristics, 7 of the 14 being resistant to non-beta-lactam antibiotics. All ampicillin-susceptible and 76% of cefotaxime-susceptible isolates had resistant mutants in the zone of inhibition when high inoculum was used. All isolates investigated had inducible chromosomal beta-lactamases, and, in addition, two isolates had an enzyme corresponding with TEM-1. Correlation of resistance patterns and antimicrobial treatment preceding bacteraemia showed that treatment with a third-generation cephalosporin was associated with beta-lactam multiresistance. In conclusion, susceptibility testing of beta-lactam antibiotics of Enterobacter must be interpreted with caution and monotherapy with an extended-spectrum cephalosporin should be avoided unless presence of resistant mutants and inducibility of beta-lactamase can be excluded.

Biotyping, phage typing, and O-serotyping of clinical isolates of Enterobacter cloacae.

The purpose of this study was to make an independent evaluation of the methods of bio-, phage-, and O-serotyping which had been used only in the laboratory of origin, and to assess the extent of possible cross-infection of Enterobacter cloacae in a Danish university hospital. The material consisted of 237 clinical isolates of E. cloacae from the clinical microbiology laboratory at Hvidovre Hospital. The typability of bio-, phage-, and serotyping was 100%, 83%, and 85%, respectively. Reproducibility of serotyping was 90% and of phage typing 96% if two major differences were allowed to differentiate between patterns. O-serotyping had the highest discriminatory power and combination of all typing methods further increased discrimination. Outbreaks of E. cloacae were not evident in clinical departments, but cross-infections from one department to another could not be completely ruled out. We concluded that the combination of bio-, phage- and O-serotyping is sufficiently discriminating and will be satisfactory in the majority of clinical situations.

Large-scale testing of women in Copenhagen has not reduced the prevalence of Chlamydia trachomatis infections.

OBJECTIVE: To examine the impact of a stable, large-scale enzyme immunoassay (EIA) Chlamydia trachomatis testing situation in Copenhagen, and to estimate the impact of introducing a genomic-based assay with higher sensitivity and specificity. METHODS: Over a five-year study period, 25 305-28 505 women screened for chlamydial infection each year, corresponding to 19.3% of the female population in Copenhagen, Denmark, were analyzed. RESULTS: The C. trachomatis age-specific examination percentage and age-specific positive percentage were unchanged during the study period. For EIA, the age-specific positive predictive value of a test decreased from 94% at age 17 to only 50% at age 34 years. Irrespective of the choice of diagnostic test, only about 30% of chlamydial infections would be diagnosed, given current strategies. CONCLUSION: Although genomic detection assays will increase the positive and negative predictive values of the Chlamydia test result, new screening strategies for both men and women in younger age groups will be necessary if chlamydial infections are to be curtailed.

Klebsiella typing: pulsed-field gel electrophoresis (PFGE) in comparison with O:K-serotyping.

OBJECTIVE: To compare pulsed-field gel electrophoresis (PFGE) typing and O:K-serotyping of Klebsiella in two different epidemiological settings. METHODS: One hundred and four bacteremia isolates without known epidemiological relation and 47 isolates from an outbreak in a neonatal intensive care unit (NICU) were K-typed by countercurrent immunoelectrophoresis (CCIE), O-typed by an inhibition enzyme-linked immunosorbent assay method, and typed by pulsed-field gel electrophoresis (PFGE) using the restriction enzyme XbaI. RESULTS: Typing data for the 104 bacteremia isolates were compared with regard to typability, number of types, maximum number of isolates per type, and the Discriminative Index (DI). O-typing combined with K-typing (DI 0.98) as O:K-serotyping (DI 0.99) gave a very discriminative typing system, whereas O-typing alone was not very discriminative (DI 0.76). PFGE (DI 1.00) was a more discriminative typing method than O:K-serotyping, as it could subdivide 13/22 O:K-serotypes into smaller groups. Isolates with the same PFGE-type had the same O:K-serotype, indicating that isolates with different O- and/or K-types could be expected to be of different PFGE-types. Typing of the 47 isolates from the outbreak in the NICU showed that 38 isolates belonged to a single clone, and that during an epidemic limited in time and space, differences in the electrophoretic patterns of up to five bands between a parental pattern type and a subtype may be found in the PFGE profiles. CONCLUSIONS: Both O:K-serotyping and PFGE typing are highly discriminative typing methods. PFGE is the most discriminative method and is excellent for typing outbreaks with few isolates. If large numbers of isolates are to be typed, a more convenient strategy might be first to K- or O:K-serotype isolates followed by PFGE typing of possible identical isolates. Since K- or O:K-serotyping is a definitive typing method, while PFGE typing is a comparative one, PFGE cannot, for the time being, replace O:K-serotyping for surveillance purposes.

Resistance problems in two university hospitals in Denmark.

In 1997, 80 and 89% of Staphylococcus aureus isolated from university hospitals in Odense and Hvidovre respectively were resistant to penicillin and 32.0 and 41% of Escherichia coli were resistant to ampicillin. There were low incidences of methicillin resistance in S. aureus (<1%), penicillin resistance in Streptococcus pneumoniae (3%),and gentamicin in E. coli (2%). These figures might reflect the low use of antibiotics in Denmark.

Bacterial contamination of the blood compartment originating from the dialysate in haemodialysers.

Micro-organisms originating from the dialysate compartment invaded the blood compartment of ' Rhodial RP6 ' haemodialysers in 16 out of 639 dialyses investigated (2.5 per cent). Analysis of the data suggested that the probable access of bacteria to the blood compartment was by way of minor defects in the dialysis membrane. The patients experienced no obvious symptoms or signs of sepsis which could be ascribed to the presence of micro-organisms in the blood compartment.

Epidemiological characterization of Klebsiella isolates from patients in a renal department.

One-hundred-and-eighteen endemic patient isolates of Klebsiella pneumoniae, collected consecutively from clinical specimens of renal patients over a 3 1/2-year period, were studied using biotypes and antimicrobial sensitivity patterns as epidemiological markers. Minor temporal clusters were demonstrable among the more commonly occurring biotypes and resistance types, and comprised isolates acquired inside and outside the renal department. Among isolates acquired in the department two clusters could be demonstrated, in peritoneal dialysate and respiratory tract specimens. Both extended over the whole observation period and occurred independently of the temporal clusters described above. They may have represented common-source outbreaks, but further elucidation was not possible with the data obtained.

A clinical and bacteriological evaluation of a re-use system for disposable haemodialysers.

Re-use of disposable haemodialysers, cleaned and disinfected with sodium hypochlorite and benzoic acid, was evaluated bacteriologically and clinically during the course of 514 dialysis treatments, performed with 206 RP6 dialysers used up to three times. Microbial contamination of the dialysers, which may be ascribed to the re-use system, was not observed. At the start of the dialyses, contamination of the dialysate compartment was lower in re-used dialysers, but during the dialyses it increased more rapidly than in dialysers used for the first time. Contamination of the blood compartment was the same with new and re-used dialysers. Clinical side effects, which may be ascribed to the re-use of dialysers, were not observed. The tested re-use system is hygienically acceptable and can be recommended for routine use, provided that the contamination of the dialysate is kept at a level below 10(4) cfu/ml and restricted to micro-organisms of low pathogenicity. A continuing bacteriological test sampling system is advisable.