Utility of oropharyngeal real-time PCR for S. pneumoniae and H. influenzae for diagnosis of pneumonia in adults.

A lack of sensitive tests and difficulties obtaining representative samples contribute to the challenge in identifying etiology in pneumonia. Upper respiratory tract swabs can be easily collected and analyzed with real-time PCR (rtPCR). Common pathogens such as S. pneumoniae and H. influenzae can both colonize and infect the respiratory tract, complicating the interpretation of positive results. Oropharyngeal swabs were collected (n = 239) prospectively from adults admitted to hospital with pneumonia. Analysis with rtPCR targeting S. pneumoniae and H. influenzae was performed and results compared with sputum cultures, blood cultures, and urine antigen testing for S. pneumoniae. Different Ct cutoff values were applied to positive tests to discern colonization from infection. Comparing rtPCR with conventional testing for S. pneumoniae in patients with all tests available (n = 57) resulted in: sensitivity 87 %, specificity 79 %, PPV 59 % and NPV 94 %, and for H. influenzae (n = 67): sensitivity 75 %, specificity 80 %, PPV 45 % and NPV 94 %. When patients with prior antimicrobial exposure were excluded sensitivity improved: 92 % for S. pneumoniae and 80 % for H. influenzae. Receiver operating characteristic curve analysis demonstrated for S. pneumoniae: AUC = 0.65 (95 % CI 0.51-0.80) and for H. influenzae: AUC = 0.86 (95 % CI 0.72-1.00). Analysis of oropharyngeal swabs using rtPCR proved both reasonably sensitive and specific for diagnosing pneumonia caused by S. pneumoniae and H. influenzae. This method may be a useful diagnostic adjunct to other methods and of special value in patients unable to provide representative lower airway samples.

Epidemiology of penicillin-non-susceptible pneumococci in Iceland, 1995-2010.

OBJECTIVES: The first penicillin-non-susceptible pneumococci (PNSP) were identified in Iceland in 1988. A rapid increase followed, associated with expansion of a single multiresistant clone, Spain(6B)-2, peaking at 19.8% in 1993. After interventions led to reduced antimicrobial use in children, the prevalence of PNSP decreased until 1995. The aim of this study was to follow the evolution of PNSP from 1995 to 2010, the period preceding the introduction of conjugated pneumococcal vaccines into the vaccination programme. METHODS: The laboratory at the Landspitali University Hospital serves ∼ 85% of the Icelandic population. All pneumococci isolated from 1995 to 2010 (n = 13,937) were stored (-80 °C). Oxacillin-resistant isolates were serotyped and penicillin MICs were determined. Selected strains were genotyped by PFGE and multilocus sequence typing. RESULTS: In 1995, the rate of PNSP was 24.2%, declining to 13.6% in 2001, and then increasing to 38.6% in 2010. Similar changes were observed for resistance to erythromycin and tetracycline. In 1995, 60.7% of PNSP were serotype 6B, mainly the Spain(6B)-2 clone, declining to 5.7% in 2010. PNSP of serotype 19F rapidly increased after 2004 to comprise 85.8% of all serogrouped PNSP in 2010, with most isolates belonging to a single multiresistant PFGE clone identified as sequence type (ST) 271 and ST1968, representing single- and double-locus variants of the international clone Taiwan(19F)-14, respectively. PNSP were most common among young children, from the nasopharynx, middle ear and lower respiratory tract. CONCLUSIONS: The epidemiology of PNSP was dominated by two multiresistant clones. The second expanded rapidly when the first one was disappearing, causing higher antibiotic resistance rates among pneumococci than seen before in Iceland.

Invasive infections due to Streptococcus pyogenes: seasonal variation of severity and clinical characteristics, Iceland, 1975 to 2012.

Epidemiology and clinical characteristics of invasive Group A streptococcal infections (IGASI) are highly variable. Long-term studies are needed to understand the interplay between epidemiology and virulence. In a population-based study of IGASI in Iceland from 1975 to 2012, 288 cases were identified by positive cultures from normally sterile body sites. Charts were reviewed retrospectively and emm-types of viable Streptococcus pyogenes isolates (n=226) determined. Comparing the first and last decade of the study period, IGASI incidence increased from 1.09 to 3.96 cases per 100,000 inhabitants per year. The most common were emm types 1 (25%), 28 (11%) and 89 (11%); emm1 strains were most likely to cause severe infections. Infections in adults were significantly more likely to be severe during the seasonal peak from January to April (risk ratio: 2.36, 95% confidence interval: 1.34­4.15). Significant seasonal variability in severity was noted among patients with diagnosis of sepsis, respiratory infection and cellulitis, with 38% of severe infections in January to April compared with 16% in other months (p<0.01). A seasonal increase in severity of IGASI suggested that generalised seasonal increase in host susceptibility, rather than introduction of more virulent strains may play a role in the pathogenesis of these potentially fatal infections.

International links between Streptococcus pneumoniae vaccine serotype 4 sequence type (ST) 801 in Northern European shipyard outbreaks of invasive pneumococcal disease.

BACKGROUND: Pneumococcal disease outbreaks of vaccine preventable serotype 4 sequence type (ST)801 in shipyards have been reported in several countries. We aimed to use genomics to establish any international links between them. METHODS: Sequence data from ST801-related outbreak isolates from Norway (n = 17), Finland (n = 11) and Northern Ireland (n = 2) were combined with invasive pneumococcal disease surveillance from the respective countries, and ST801-related genomes from an international collection (n = 41 of > 40,000), totalling 106 genomes. Raw data were mapped and recombination excluded before phylogenetic dating. RESULTS: Outbreak isolates were relatively diverse, with up to 100 SNPs (single nucleotide polymorphisms) and a common ancestor estimated around the year 2000. However, 19 Norwegian and Finnish isolates were nearly indistinguishable (0-2 SNPs) with the common ancestor dated around 2017. CONCLUSION: The total diversity of ST801 within the outbreaks could not be explained by recent transmission alone, suggesting that harsh environmental and associated living conditions reported in the shipyards may facilitate invasion of colonising pneumococci. However, near identical strains in the Norwegian and Finnish outbreaks does suggest that transmission between international shipyards also contributed to those outbreaks. This indicates the need for improved preventative measures in this working population including pneumococcal vaccination.

The impact of domestic travel on estimating regional rates of human campylobacteriosis.

Residential locations of cases are often used as proxy measures for the likely place of exposure and this assumption may result in biases affecting both surveillance and epidemiological studies. This study aimed to describe the importance of domestic travel in cases of human campylobacteriosis reported during routine surveillance in Iceland from 2001 to 2005. Various measures of disease frequency were calculated based upon the cases' region of residence, adjusting location of domestic travel cases to their travel region, as well as separate estimations for travellers and non-travellers. Of the 376 cases included in the analysis, 37% had travelled domestically during their incubation period. Five of the eight regions were identified as high-risk when considering domestic travel whereas there were no high-risk regions when considering only region of residence. The change in regional representation of disease occurrence indicates the importance of collecting domestic travel information in ongoing surveillance activities.

Risk factors and outcome in ICU-acquired infections.

BACKGROUND: Nosocomial infections are common in intensive care units (ICU). The objectives of this study were to determine risk factors of ICU-acquired infections, and potential mortality attributable to such infections. METHODS: An observational study was performed in a 10-bed multidisciplinary ICU. For a period of 27 months, all patients admitted for >or=48 h were included. Infections were diagnosed according to Centers for Disease Control and Prevention definitions. Airway colonization was explored by molecular typing. Risk factors for infection were determined by multivariable logistic regression. Survival was analyzed with time-varying proportional hazards regression. RESULTS: Of 278 patients, 81 (29%) were infected: urinary tract infections in 39 patients (14%), primary bloodstream infections in 25 (9%), surgical site infections in 22 (8%) and pneumonia in 21 (8%). Of the total of 147 episodes, Gram-negative bacilli were isolated in 90, Gram-positive cocci in 49 and Candida sp. in 25. Risk factors for pneumonia were mechanical ventilation [odds ratio (OR=7.9, CI 1.8-35), lack of enteral nutriment (OR=8.0, CI 1.4-45) and length of time at risk (OR=1.8, CI 1.2-2.8), while gastric acid inhibitors did not affect the risk (OR=0.99, CI 0.32-3.0). Transmission of bacteria from the stomach to the airway was not confirmed. The risk of death was increased as patients were infected with pneumonia [hazard ratio (HR)=3.6; CI: 1.6-8.1], or primary bloodstream infection (HR=2.5; CI: 1.2-5.4), independent of age and disease severity. CONCLUSIONS: Mortality was increased by ICU-acquired pneumonia and primary bloodstream infections. Our findings did not support the gastro-pulmonary hypothesis of ICU-acquired pneumonia. The proposition that blood transfusions increase the risk of ICU-acquired nosocomial infections was not supported.

Emergence and spread of vancomycin resistance among enterococci in Europe.

Nowadays, six types of acquired vancomycin resistance in enterococci are known; however, only VanA and to a lesser extent VanB are widely prevalent. Various genes encode acquired vancomycin resistance and these are typically associated with mobile genetic elements which allow resistance to spread clonally and laterally. The major reservoir of acquired vancomycin resistance is Enterococcus faecium; vancomycin-resistant Enterococcus faecalis are still rare. Population analysis of E. faecium has revealed a distinct subpopulation of hospital-acquired strain types, which can be differentiated by molecular typing methods (MLVA, MLST) from human commensal and animal strains. Hospital-acquired E. faecium have additional genomic content (accessory genome) including several factors known or supposed to be virulence-associated. Acquired ampicillin resistance is a major phenotypic marker of hospital-acquired E. faecium in Europe and experience has shown that it often precedes increasing rates of VRE with a delay of several years. Several factors are known to promote VRE colonisation and transmission; however, despite having populations with similar predispositions and preconditions, rates of VRE vary all over Europe.

Control of electronic transport in graphene by electromagnetic dressing.

We demonstrated theoretically that the renormalization of the electron energy spectrum near the Dirac point of graphene by a strong high-frequency electromagnetic field (dressing field) drastically depends on polarization of the field. Namely, linear polarization results in an anisotropic gapless energy spectrum, whereas circular polarization leads to an isotropic gapped one. As a consequence, the stationary (dc) electronic transport in graphene strongly depends on parameters of the dressing field: A circularly polarized field monotonically decreases the isotropic conductivity of graphene, whereas a linearly polarized one results in both giant anisotropy of conductivity (which can reach thousands of percents) and the oscillating behavior of the conductivity as a function of the field intensity. Since the predicted phenomena can be observed in a graphene layer irradiated by a monochromatic electromagnetic wave, the elaborated theory opens a substantially new way to control electronic properties of graphene with light.

Changing epidemiology of group B streptococcal infections among adults in Iceland: 1975-2014.

We studied the bacterial characteristics and incidence of invasive infections caused by group B streptococci (GBS) in adults in Iceland in 1975-2014. A total of 145 isolates were characterized by serotyping, antimicrobial susceptibility, multilocus sequence typing and surface protein gene profiling. Disease incidence increased during the studied period (p <0.001), reaching 2.17 cases/100 000 person-years in 2013-14. Overall, serotype Ia was the most frequently found (23%), but serotypes Ib, II, III and V showed similar prevalence (14%-17%). Although there were notable changes in the proportion of most serotypes during the study period, only the decline of serotype III was statistically supported (p = 0.003) and was reflected in a decrease of clonal complexes CC17 and CC19 that included most serotype III isolates (p <0.04). On the other hand, the increase in frequency of CC1 was caused by two lineages expressing distinct serotypes: ST1/V/alp3 and ST196/IV/eps. Underlying the relative stability of serotype Ia were major changes in the lineages expressing this serotype, with an increase in the relative importance of CC23, including both ST23/Ia/eps and ST24/Ia/bca lineages, and a decrease in CC7. Nine cases of invasive GBS disease were caused by ST7, of possible zoonotic origin. All isolates were susceptible to penicillin. Rates of erythromycin and clindamycin resistance were 8.3% and 9.7%, respectively. An over-representation of resistance solely to clindamycin was associated with the unusual lsaC gene and serotype III ST19/rib lineage (p <0.001).

Nationwide study of recurrent invasive pneumococcal infections in a population with a low prevalence of human immunodeficiency virus infection.

Recurrent invasive infections caused by Streptococcus pneumoniae are rare, and often considered to be indicative of serious underlying illness. However, the prevalence of this problem, and the relevance of specific predisposing conditions, can be hard to assess, since many of the studies are based on specific risk groups. A population-based study of recurrent invasive pneumococcal disease in Iceland during the 30-year period 1975-2004 was performed. Clinical information, including mortality and vaccine use, was analysed retrospectively. Invasive pneumococcal isolates were serotyped and susceptibility testing was performed. During this period, 36 (4.4%) of 819 patients who survived an initial infection experienced recurrence, with a median time between episodes of 9.7 months. Pneumonia with bacteraemia was the most common clinical diagnosis (48% of cases), followed by bacteraemia without a clear focus (21%) and meningitis (13%). Most (94%) of the patients had identifiable predisposing conditions, most commonly, multiple myeloma in adults, and antibody deficiencies in children. Compared with children, adults were more likely to present with pneumonia (65% vs. 18%; p 0.0001). No significant change in the 30-day mortality rate was observed during the three decades of the study. Only 26% of eligible patients received pneumococcal vaccination. Patients with recurrent invasive pneumococcal disease should be investigated thoroughly for underlying diseases. Greater use of pneumococcal vaccines should be encouraged among high-risk patients. More effective preventive and therapeutic measures are needed to improve outcomes.

Helicobacter pylori antibodies and gastric cancer in Iceland - The decline in IgG antibody level is a risk factor.

H. pylori infection is considered a causal agent of duodenal ulcer and a significant risk factor for gastric cancer. Retrospective cohort studies have demonstrated a significant association between presence of antibody to H. pylori and gastric cancer when using samples obtained years before the diagnosis but not at the time of diagnosis. The present study investigates, in a population-based cohort, whether a decline occurs in H. pylori antibody levels before the diagnosis of stomach cancer. Repeat samples (2 to 5) were available from 23 persons with gastric cancer taken up to 20 years before the diagnosis and 128 control subjects matched for gender, age, time and number of repeat samples. The odds ratio of developing stomach cancer was 1.16 (95% CI 1.05-1.28) for those showing decline in antibody levels of 1 relative antibody activity unit per year versus those with constant or rising levels. We conclude that this decline in antibody levels in cases, and not in controls, supports an active role of H. pylori in the pathogenesis of gastric cancer by causing atrophic gastritis, and provides a better risk assessment for gastric cancer compared to single measurements.

Immune responses of infants vaccinated with serotype 6B pneumococcal polysaccharide conjugated with tetanus toxoid.

BACKGROUND: Streptococcus pneumoniae is a major cause of meningitis, bacteremia, pneumonia and otitis media. Pneumococcal polysaccharides are not immunogenic in infants, but improved immunogenicity of polysaccharide-protein conjugates has been demonstrated. Antibiotic-resistant pneumococci have increased the need for an effective vaccine. OBJECTIVE: To study the safety and immunogenicity of a pneumococcal type 6B polysaccharidetetanus toxoid conjugate (Pn6B-TT) in infants and to assess the function of antibodies. METHODS: Healthy infants were injected, Group A at 3, 4 and 6 months (n = 21) and Group B at 7 and 9 months (n = 19). Booster injection was given at 18 months. Antibodies were measured by enzyme-linked immunosorbent assay and radioimmunoassay, and functional activity was measured by opsonization of radiolabeled pneumococci. Nasopharyngeal cultures were obtained. RESULTS: No significant adverse reactions were observed. Pn6B-IgG (enzyme-linked immunosorbent assay) increased to a geometric mean of 0.62 microgram/ml (P = 0.367, compared with prevaccination titers) in Group A at 7 months and 1.22 micrograms/ml (P < 0.001) in Group B at 10 months. Total Pn6B antibodies (radioimmunoassay) were 44 ng of antibody N/ml (P < 0.053) in Group A and 211 ng of antibody N/ml (P < 0.001) in Group B. A smaller increase in IgM and IgA anti-Pn6B was observed. Reinjection at 18 months elicited booster responses in total and IgG anti-Pn6B; 62% of those in Group A and 79% of those in Group B had > 300 ng of antibody N/ml. Opsonic activity, after initial and booster vaccinations, correlated with Pn6B-antibody titers. Three infants with nasopharyngeal cultures repeatedly positive for serogroup 6 had poor serum IgG responses. CONCLUSION: Our results demonstrate that Pn6B-TT is safe, elicits functional antibodies and memory responses in infants.

Assessment of rapid methods of pneumococcal antigen detection in routine sputum bacteriology.

Sputum specimens from 480 patients were examined for the presence of pneumococci by Gram film and culture and for pneumococcal antigen by counterimmunoelectrophoresis, coagglutination, and latex agglutination. Ninety six positive specimens were detected. Gram film and culture provided the most reliable techniques in well taken specimens collected early in the illness before antibiotic treatment had started. More than 70% of the specimens examined were submitted after starting antibiotics, however, and in these specimens, methods of antigen detection proved of greater value than either Gram film or culture. Counterimmunoelectrophoresis, coagglutination, and latex agglutination were similar in sensitivity and specificity, but coagglutination and latex agglutination were much easier to perform and to read.

Effect of antimicrobial use and other risk factors on antimicrobial resistance in pneumococci.

Penicillin-resistant and multi-resistant pneumococci have spread globally and reached high prevalence in many countries. Antimicrobial use is considered a major driving force for resistance, although the influence in the community has not been as clearly demonstrated. Other risk factors may be important, and only with a clear understanding of the risk factors can effective control measures be introduced. The main habitat of the pneumococcus is the nasopharynx of children. Carriage increases from birth and is maximal at pre-school age. Antimicrobial use in children is likely to have a significant influence on the susceptibility of pneumococci. Most studies looking for risk factors for resistance in pneumococci have identified antimicrobial use as a risk factor, especially the following aspects: ongoing, recent, repeated, frequent, and prophylactic antibiotic use. The effect of individual classes of antimicrobials has not been studied in detail but use of beta-lactam antibiotics and trimethoprim-sulpha has been associated with increased risk. Other risk factors are young age, nosocomial acquisition, prior hospitalization, and HIV infection. Day-care centers can facilitate the spread of resistant pneumococci and an Icelandic study showed that carriage of resistant pneumococci was associated with young age, domicile in an area with high antimicrobial consumption, recent antimicrobial use, frequent antimicrobial use, and use of trimethoprim-sulpha. The rapid increase of penicillin-resistant pneumococci in Iceland was met with propaganda against overuse of antimicrobials, which lead to reduction of antimicrobial use and subsequently a reduced incidence of penicillin-resistant pneumococci. This reduction may be related to reduced antimicrobial use. Reducing antimicrobial use should be considered important for programs aimed at reducing antimicrobial resistance.

Epidemiology of penicillin resistant pneumococci in Iceland.

The prevalence of penicillin-resistant pneumococci (PRP) has been increasing, with the highest levels reported from countries with relatively unrestricted antimicrobial use. It has been low in northern Europe except Iceland, which is disconcerting as antimicrobial use in Iceland has been relatively restricted. This suggests that other factors may facilitate their spread. By studying their epidemiology and possible risk factors for carriage, we have attempted to explain their rapid spread in Iceland. The incidence of infections caused by PRP (as percentage of infections considered due to pneumococci) has increased from 0% in 1988 to 2.3% in 1989, 2.7% in 1990, 8.4% in 1991, 16.3% in 1992, and 19.8% in 1993. The infections have mainly affected 0- to 3-year-old children (71.4%), and the PRP belonged almost exclusively to serogroups 6, 19, and 23 (98.8%). Most were serotype 6B multiresistant (75%; resistant to penicillin (MIC = 1.0), cephalothin, erythromycin, clindamycin, tetracycline, chloramphenicol, fusidic acid, sulfonamides, and trimethoprim), and believed to belong to a single clone originating from Spain. The PRP have been prevalent in healthy children, 0-10% nasopharyngeal carriage, especially in day-care centers, with the highest prevalence in areas that had the highest antimicrobial consumption. Recent antimicrobial consumption, especially of trimethoprim-sulfa, appeared to increase PRP carriage. The rapid spread of PRP in Iceland may have been facilitated by high antimicrobial consumption in day-care centers (especially of trimethoprim-sulfa) which are attended by the majority of Icelandic children.

Carriage of respiratory tract pathogens and molecular epidemiology of Streptococcus pneumoniae colonization in healthy children attending day care centers in Lisbon, Portugal.

In an effort to establish the rate of carriage of antibiotic resistant respiratory pathogens in children attending urban day care centers (DCC) in Portugal, seven DCC in Lisbon were selected for determining the rate of nasopharyngeal colonization of children between the ages of 6 months to 6 years by Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis. Of the 586 children studied between January and March 1996, 47% carried S. pneumoniae, 72% H. influenzae, and 54% M. catarrhalis. Twenty-four percent of the pneumococci had reduced susceptibility to penicillin, and most of these belonged to serogroups 19, 23, 14, and 6. An additional 19% were fully susceptible to penicillin but showed decreased susceptibility to other antimicrobials. These isolates expressed serogroups 6, 11, 14, 18, 19, and 34. The majority (96%) of M. catarrhalis and 20% of H. influenzae were penicillin resistant due to the production of beta-lactamases. Recent antimicrobial use was associated with carriage of penicillin non-susceptible pneumococci and beta-lactamase producing H. influenzae (p < 0.05). Individual DCC differed substantially from one another in their rates of carriage of antibiotic resistant H. influenzae and S. pneumoniae. Characterization of antibiotic resistant S. pneumoniae isolates by molecular fingerprinting techniques showed that each DCC had a unique microbiological profile, suggesting little, if any, exchange of the resistant microbial flora among them. An exception to this was the presence of isolates belonging to two internationally spread epidemic clones: the multiresistant Spanish/USA clone expressing serotype 23F, and the penicillin and sulfamethoxazole-trimethoprim resistant French/Spanish clone (serotype 14) which were detected in four and three DCC, respectively.

Adherence of staphylococci to intravascular catheters.

Adherence of seven strains of Staphylococcus epidermidis and three strains of S. aureus to three types of intravascular catheters was assessed by ATP bioluminescence, by culture after ultrasonication and by scanning electronmicroscopy. The catheter materials studied were silicone elastomer, thermoplastic polyurethane and polyurethane coated with Hydromer, a coating which absorbs water and provides a hydrophilic sheath around the catheter. The adherence assays were performed in phosphate-buffered saline on a rotary shaker at 37 degrees C, with the catheters precoated with serum and uncoated, and the results were correlated with bacterial hydrophobicity. There was wide strain-to-strain variation in bacterial adherence; S. aureus and slime-producing S. epidermidis strains adhered better than did non-slime-producing strains. Overall, there was less bacterial adherence to Hydromer-coated catheters than to polyurethane and silicone catheters but it was unrelated to bacterial hydrophobicity. Serum coating of catheters resulted in marked reduction of bacterial adherence.

The role of extracellular slime in opsonophagocytosis of Staphylococcus epidermidis.

Infections caused by coagulase-negative staphylococci (CNS) are a major problem in immunocompromised patients. It has been claimed that extracellular slime production by CNS predicts pathogenicity and inhibits host defences. Luminol-enhanced neutrophil chemiluminescence (CL) and bacterial killing assays were used to assess the effect of slime production on opsonophagocytosis and killing by polymorphonuclear leucocytes in vitro. There was wide variation in CL induction amongst the 43 strains of Staphylococcus epidermidis examined. The presence of slime had no influence either on the requirement or on the efficiency of opsonisation. Slime-producing and non-slime-producing strains showed a stepwise increase in induced CL up to a serum concentration of 10%, and were dependent on complement for efficient phagocytosis. The bacterial killing assays confirmed the CL results. Our data suggest that extracellular staphylococcal slime has no specific anti-opsonic property in vitro. Opsonophagocytosis may still be hampered in vivo by the physical presence of slime.

Cultural and electronmicroscopic studies of the effect of penicillin on tolerant oral streptococci.

Penicillin-tolerant and -sensitive strains of oral streptococci were treated with penicillin to determine the production of a post-antibiotic effect (PAE). No PAE was seen with any of the S. sanguis strains tested but most strains of the other oral streptococcal species produced a PAE. Cultures on nitrocellulose filters treated with penicillin were examined by scanning electronmicroscopy and showed that tolerant and sensitive strains lost the ability to adhere to the filter after application of antibiotic. When the filters were treated with beta-lactamase, before processing for microscopy, the tolerant strains but not the sensitive ones recovered and grew in a confluent lawn similar to the control cultures that had not received antibiotic. Transmission electronmicroscopic examination of similarly treated cultures produced comparable results. Bizarre morphological changes were a feature of the tolerant strains of S. sanguis.