Rapid selection of complement-inhibiting protein variants in group A Streptococcus epidemic waves.

Serotype M1 group A Streptococcus strains cause epidemic waves of human infections long thought to be mono- or pauciclonal. The gene encoding an extracellular group A Streptococcus protein (streptococcal inhibitor of complement) that inhibits human complement was sequenced in 1,132 M1 strains recovered from population-based surveillance of infections in Canada, Finland and the United States. Epidemic waves are composed of strains expressing a remarkably heterogeneous array of variants of streptococcal inhibitor of complement that arise very rapidly by natural selection on mucosal surfaces. Thus, our results enhance the understanding of pathogen population dynamics in epidemic waves and infectious disease reemergence.

Localized adherence by enteropathogenic Escherichia coli is an inducible phenotype associated with the expression of new outer membrane proteins.

Enteropathogenic Escherichia coli grow as discrete colonies on the mucous membranes of the small intestine. A similar pattern can be demonstrated in vitro; termed localized adherence (LA), it is characterized by the presence of circumscribed clusters of bacteria attached to the surfaces of cultured epithelial cells. The LA phenotype was studied using B171, an O111:NM enteropathogenic E. coli (EPEC) strain, and HEp-2 cell monolayers. LA could be detected 30-60 min after exposure of HEp-2 cells to B171. However, bacteria transferred from infected HEp-2 cells to fresh monolayers exhibited LA within 15 min, indicating that LA is an inducible phenotype. Induction of the LA phenotype was found to be associated with de novo protein synthesis and changes in the outer membrane proteins, including the production of a new 18.5-kD polypeptide. A partial NH2-terminal amino acid sequence of this polypeptide was obtained and showed it to be identical through residue 12 to the recently described bundle-forming pilus subunit of EPEC. Expression of the 18.5-kD polypeptide required the 57-megadalton enteropathogenic E. coli adherence plasmid previously shown to be required for the LA phenotype in vitro and full virulence in vivo. This observation, the correspondence of the 18.5-kD polypeptide to an EPEC-specific pilus protein, and the temporal correlation of its expression with the development of the LA phenotype suggest that it may contribute to the EPEC colonial mode of growth.

Evaluation of a commercial MRSA assay when multiple MRSA strains are causing epidemics.

Rapid and reliable diagnostic methods are needed to control methicillin-resistant Staphylococcus aureus (MRSA) transmission. We studied the BD GeneOhm MRSA Assay which is based on one specific amplification product at the junction of the right extremity sequence of the staphylococcal cassette chromosome mec (SCCmec) and the chromosomal sequence of orfX of S. aureus. The test was applied on 95 clinical isolates in Finland: 83% were positive. The isolates giving negative results represented several pulsed-field gel electrophoresis (PFGE) types and harboured SCCmec types IV, V, VI or were new types with different combinations of ccr genes.

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.

Eradication of methicillin-resistant Staphylococcus aureus from a health center ward and associated nursing home.

BACKGROUND: Long-term health care facilities have been recognized as reservoirs of multiresistant bacterial strains, especially methicillin-resistant Staphylococcus aureus (MRSA). Efforts to control MRSA in this setting usually have been only partially effective. We describe herein the eradication of epidemic MRSA from a Finnish health care center ward and affiliated nursing home. METHODS: The methods to control MRSA included (1) contact isolation precautions, (2) screening for asymptomatic carriage, (3) eradication of carriage, and (4) education of staff on hygienic measures. The first 6 patients with MRSA-positive findings were referred without delay to the Infectious Diseases Unit of the adjacent university hospital for eradication treatment. Later, an isolation unit of 6 rooms was founded in the health care center, where the MRSA-colonized patients were nursed as a separate cohort until they, in succession, were referred to the Infectious Diseases Unit for decolonization. RESULTS: From May 20 through August 17, 1993, the epidemic MRSA strain was isolated from 8 long-term patients on the 40-bed ward of the health care center, 4 of the 59 residents of the nursing home, and 1 member of the staff. Eradication of carriage was successful in all except 1 patient with dementia, who was nursed in contact isolation in the health care center until his death 21 months later. CONCLUSIONS: It is possible to eradicate MRSA from a long-term health care facility even after 13 cases by applying strict control measures. Our experience may be valuable in the future decision-making process for control of new and more challenging multiresistant bacteria, eg, vancomycin-resistant strains of MRSA.

The epidemiology of severe Streptococcus pyogenes associated disease in Europe.

Several European countries reported outbreaks of severe disease caused by Streptococcus pyogenes in the late 1980s. This marked a departure from the previous decades, where very few such outbreaks were noted. These changes in disease occurrence formed part of a global phenomenon, the reasons for which have yet to be explained. Results of surveillance activities for invasive S. pyogenes infection within Europe over the past fifteen years identified further increases in many countries. However, variations in surveillance methods between countries preclude robust comparisons being made, illustrating the need for a unified surveillance strategy across Europe. This was finally embodied in the Strep-EURO programme, introduced in 2002.

The epidemiology of severe Streptococcus pyogenes associated disease in Europe.

Several European countries reported outbreaks of severe disease caused by Streptococcus pyogenes in the late 1980s. This marked a departure from the previous decades, where very few such outbreaks were noted. These changes in disease occurrence formed part of a global phenomenon, the reasons for which have yet to be explained. Results of surveillance activities for invasive S. pyogenes infection within Europe over the past fifteen years identified further increases in many countries. However, variations in surveillance methods between countries preclude robust comparisons being made, illustrating the need for a unified surveillance strategy across Europe. This was finally embodied in the Strep-EURO programme, introduced in 2002.

Cell surface properties of Acinetobacter baumannii.

Cell surface properties of 78 strains of Acinetobacter baumannii of different origin (lower respiratory tract, wound, blood and environment) were investigated. The bacterial adhesion to collagen, fibronectin, fibrinogen and vitronectin was detected by particle agglutination assays. Salt aggregation tests were used to determine the cell surface hydrophobicity of isolated A. baumannii strains. We found that A. baumannii strains originating from patients with wound infection and bacteraemia showed significantly lower aggregative properties compared to respiratory and environmental strains. Electron microscopic investigations revealed more fimbriated bacterial cells among the highly aggregative A. baumannii strains. This study demonstrates that the investigated A. baumannii strains can be divided into two different groups according to their cell surface properties and source of isolation, whereas the majority of strains, from the lower respiratory tract and the hospital environment expressed strong adhesive properties.

Neonatal group B streptococcal disease in Finland: a ten-year nationwide study.

BACKGROUND: Group B Streptococcus (GBS) is the most common cause of invasive infections in newborns. GBS bacteria are typed on the basis of capsular polysaccharides or surface-localized proteins. Both polysaccharides and protein antigens have been suggested as potential vaccine candidates. METHODS: A prospective nationwide laboratory-based study of invasive GBS infections in children younger than 3 months of age was conducted in 1985 through 1994. Isolates were serotyped by immunodiffusion in agar gel with HCl extracts and rabbit antisera. Clinical diagnoses and case fatalities were verified from the patient records or the national hospital discharge register. RESULTS: There were 485 cases registered during the 10-year period. The incidence of disease was 0.76/1000 live births. The case fatality rate was 8.0%. Of the 485 cases 398 (83%) were early onset and 87 (17%) late onset infections. The most common clinical diagnosis was bacteremia (77%) without an identified focus of infection. Other diagnoses included meningitis (17%), pneumonia (3%), osteomyelitis or septic arthritis (2%), pyelonephritis or cellulitis. Serotyping of 395 isolates revealed that 47% were of serotype III or III/R, 23% of Ia/c, 11% of Ib, 6% of II/R, 8% of IV, 1% of V and 7% were nontypable. CONCLUSIONS: The clinical picture of GBS disease and serotype distribution are similar to what has been reported from other countries. Serotypes III and III/R dominated (47% of all infections), especially in late onset disease. On the basis of these results a GBS vaccine including at least the Ia, Ib, II and III components would provide coverage against 88% of GBS serotypes causing neonatal disease in Finland.

Neonatal Candida parapsilosis outbreak with a high case fatality rate.

A Candida parapsilosis outbreak of 58 cases in a neonatal intensive care unit lasted for 55 months. Patients infected by or colonized with C. parapsilosis were mainly very low birth weight infants (birth weight < 1500 g). Their mean birth weight was 817 g and their mean gestational age was 28 weeks. Statistical analysis including logistic regression confirmed that prematurity was the main risk factor. The analysis also suggested that C. parapsilosis infection (or colonization) was associated with a poor prognosis. In infants with gestational age < 29 weeks the risk for death in C. parapsilosis-infected patients was 16-fold greater than in those with no C. parapsilosis infection. The case fatality rate of C. parapsilosis patients was higher than that of the controls (9 of 23 vs. 1 of 40; P < 0.0001). The outbreak was most likely a result of cross-infection because C. parapsilosis could be isolated only from the patients and from the hands of four nurses immediately after they had cared for a colonized patient. Cessation of the outbreak was temporally associated with long term parenteral fluconazole (6 mg/kg/day) prophylaxis.

Bacterial adherence to self-reinforced polyglycolic acid and self-reinforced polylactic acid 96 urological spiral stents in vitro.

The aim of this study was to evaluate the bacterial adherence to biodegradable self-reinforced polyglycolic acid (SR-PGA) and self-reinforced poly-DL-lactic acid (SR-PLA 96) spiral stents in vitro. They are used as temporary urethral stents in urology. Gold-plated metal wire, polyurethane and latex were used as controls. Materials were incubated up to 28 days in artificial urine, after which a bacterial suspension was added. After detaching by sonication the adhesive bacteria were analysed as colony forming units (CFUs) and by scanning electron microscopy (SEM) analysis. Adhesion was more significantly correlated to stent bacterial type than to the tested material in both assays. No encrustation was seen on SR-PGA or SR-PLA 96. SR-PGA and SR-PLA 96 had no effect on the bacterial growth. In conclusion, the bacterial properties are equally or more important than the material properties in the adhesion process.

Experimental Escherichia coli peritonitis in immunosuppressed mice: the role of specific and non-specific immunity.

An experimental Escherichia coli septicaemia-peritonitis model was adapted to immunosuppressed mice. The mice were made neutropenic by a sublethal dose of cyclophosphamide, which resulted in a 100-fold increase in their susceptibility to intraperitoneal injection of E. coli O18:K1. A lethal infection could be prevented by passive immunisation with anti-K1 capsular or anti-O18 LPS antibodies but not with anti-J5 bacterial antibodies. The anti-K1 and anti-O18 antisera were able to increase the LD50 of the E. coli challenge by factors of 50 and 5, respectively. The role of non-specific, lipopolysaccharide (LPS)-mediated resistance to infection was also investigated in this model, in which only long-living phagocytic cells such as macrophages are believed to be functional. Pretreatment of mice with LPS was shown to prevent growth of the bacterial challenge in the peritoneal cavity and blood and to result in a five-fold increase in the LD50 of the challenge strain. These findings suggest an important role for macrophages as effector cells in defence against E. coli infection.

Killing of Escherichia coli in the peritoneal cavity of convalescent mice; role of specific and non-specific immune mechanisms.

Mice surviving a sublethal E. coli O18:K1 infection possess a greatly increased resistance to a subsequent lethal E. coli O18:K1 peritonitis. A similar increase in resistance can also be achieved by LPS pretreatment. The early course of the infection in the convalescent mice at day 1 was identical to that in LPS-pretreated mice. At this time, the convalescent mice were also able to restrict the growth of the heterologous E. coli O78(ColV) strain, suggesting that non-specific phagocyte activation was responsible for the increased destruction of the bacteria. At day 4, the kinetics of infection in convalescent mice were identical to those in mice passively immunised with specific anti-K1 capsule antiserum. A rapid decline in the numbers of viable homologous, but not heterologous, bacteria took place in the peritoneal cavity suggesting effective antibody-mediated opsonisation followed by phagocytosis and killing of the bacteria.

Protective capacity of antibodies to outer-membrane components of Escherichia coli in a systemic mouse peritonitis model.

Antibody-mediated protection was studied in an experimental murine model of peritonitis-septicaemia with Escherichia coli O18:K1. Protection from lethal intraperitoneal challenge was achieved by passive immunisation with horse anti-K1 capsular antiserum (H46) or rabbit antiserum to the homologous O18 antigen. The maximum increase in LD50 achieved with anti-K1 and anti-O18 antibodies was 10- and 5-fold, respectively. The protective capacity of the anti-O serum was found to be in the IgG fraction. Rabbits were also immunised with various semi-purified or purified outer-membrane-protein preparations (porins and OmpA protein) from rough E. coli or Salmonella strains or with whole E. coli J5 bacteria. Although this immunisation resulted in high antibody titres to homologous and, to a lesser extent, also to heterologous antigens, none of the antisera protected against challenge with the capsulate E. coli O18:K1 bacteria.

Lipopolysaccharide-induced non-specific resistance to systemic Escherichia coli infection in mice.

A high degree of non-specific resistance to a lethal systemic Escherichia coli infection was induced in mice by pretreatment with a small dose (less than 5 micrograms/mouse) of the homologous lipopolysaccharide (LPS) or with heterologous rough-type LPS from E. coli K-12. The route of LPS administration, intraperitoneally or subcutaneously, did not influence the development of resistance, suggesting that a systemic cell activation was responsible for the effect. The enhanced elimination of bacteria was similar to that in mice recovering from a sublethal E. coli infection. In the LPS-treated mice, elimination of the challenge bacteria from the peritoneal cavity and the blood started 3-4 h after challenge whereas, in controls, the bacterial numbers continued to increase until the mice died. The detoxified LPS derivative, monophosphoryl lipid A (MPL), also increased the survival of mice infected with E. coli O18:K1. However, the dose of MPL required for optimal infection resistance was 100-fold greater than that of native, E. coli K-12 LPS, corresponding to the 100-fold reduced toxicity of MPL for mice and rabbits in lethality and pyrogenicity assays.

Biological characterization of Campylobacter fetus lipopolysaccharides.

Lipopolysaccharides (LPS) of three strains of Campylobacter fetus (subspp. fetus and venerealis, and serotypes A and B), a bacterium of veterinary importance but also a cause of various infections in humans, were assessed for their ability to induce mitogenicity, gelation of Limulus amebocyte lysate, lethal toxicity in mice, and pyrogenicity in rabbits. All C. fetus LPS exhibited activities lower than those of Salmonella typhimurium LPS. LPS of C. fetus subsp. fetus serotype A had the lowest activity in all assays. Since the majority of C. fetus subsp. fetus isolates from humans are serotype A, the lower biological activities of this LPS may aid the pathogenesis of such strains. The lower activities of C. fetus LPS compared with those of S. typhimurium LPS may reflect the presence of longer fatty acid chains in the lipid A of C. fetus LPS, whereas interstrain differences in C. fetus LPS bioactivities may be related to some property influenced by composition of the saccharide moiety.

Outbreak caused by two multi-resistant Acinetobacter baumannii clones in a burns unit: emergence of resistance to imipenem.

Since early 1992 an increased number of tobramycin- and imipenem-resistant Acinetobacter spp. were observed causing colonization, wound infections, and bacteraemias in a burns and plastic surgery unit. This raised the question of whether this outbreak was caused by a single or by multiple Acinetobacter spp. clones. To study this, 97 Acinetobacter spp. isolates from clinical samples from different hospital units as well as isolates from the environment and the hands of the staff were characterized by antibiogram, plasmid profile and ribotyping. Two dominant multi-resistant A. baumannii clones were identified; one of them was sensitive to polymyxin B only. There was a close correlation between the results obtained by plasmid profiling and ribotyping. No common environmental source or significant hand carriage, or spread of these strains outside the unit were detected. The burns patients were the most likely reservoir, and strain transmission occurred in spite of strict control measures.

Changing epidemiology of methicillin-resistant Staphylococcus aureus in Finland.

Data on methicillin-resistant Staphylococcus aureus (MRSA) cases notified to the National Infectious Disease Register (NIDR) and antibiotic resistance profiles of MRSA isolates sent to the national reference laboratory between 1997 and 2002 were analysed. In addition, the diagnostic methods used for MRSA identification in Finnish microbiology laboratories, the number of MRSA screening specimens studied, and the MRSA situation in long-term care facilities in 2001 were reviewed. MRSA cases notified to the NIDR rose from 120 in 1997 to 597 in 2002 (from 2.3 to 11.5 cases per 100,000 population). The increase was greatest in elderly people and outside Helsinki metropolitan area, in the districts where the proportion of non-multiresistant strains was most prominent. The National Committee for Clinical Laboratory Standard's guidelines for the oxacillin disk diffusion test were followed, except for the incubation temperature and time, which may have hindered detection of some MRSA strains. There was a wide geographic variation in the rates of MRSA, but this was not related to screening activity. MRSA isolates from long-term facilities accounted for more than half of the notifications to the NIDR in 2001.

Eradication of the long-term carriage of methicillin-resistant Staphylococcus aureus in patients wearing dentures: a follow-up of 10 patients.

To cure the long-term carriage of methicillin-resistant Staphylococcus aureus (MRSA), eradication treatment was given to 10 patients wearing complete dentures. In addition to multiple body sites, MRSA was cultured from the dentures of six patients. The contaminated dentures were rebased and sterilized with heat in order to prevent recolonization. The patients did not use their dentures during the decolonization therapy. Subsequently, MRSA was eradicated from three of these patients but three others remained MRSA-positive despite at least two courses of combined systemic and topical eradication treatment. These particular patients had persistent stomatitis and their dentures were a poor fit, in poor condition and repeatedly grew MRSA. Eradication treatment was successful in the remaining four patients whose dentures were MRSA-negative. These results confirm that dentures may function as foreign bodies and sustain persistent nasopharyngeal. MRSA colonization. Therefore, we suggest that whenever eradication of MRSA is deemed necessary in cases of nasal, oral or pharyngeal carriage, heat treatment of the dentures should be included. Further comparative studies with larger patient populations are needed to evaluate the contribution of dentures to the long-term carriage of MRSA, as well as to assess the value of denture sterilization during the eradication course.

Control of methicillin-resistant Staphylococcus aureus outbreak involving several hospitals.

This population-based, retrospective, cohort study describes a large methicillin-resistant Staphylococcus aureus (MRSA) epidemic caused by one strain (E1) in the greater Helsinki region. The epidemic comprised 210 cases at several hospitals, but was finally controlled. The study period ranged from June 1991 to December 2000. The epidemic peaked in 1993-1995 with 143 cases (68% of total cases). From August 1993, all MRSA-positive cases at the eight municipal hospitals were isolated and barrier nursed. Contacts were cohorted and screened for MRSA colonization. Decolonization treatment was administered to some chronic carriers. MRSA cases and contacts were identified in the joint patient register of the municipal hospitals from August 1993. The annual incidence of MRSA E1 in Helsinki City area per 100,000 inhabitants rose from 0.2 in 1991 to 13.6 in 1994. It decreased from 1995, reaching 0.7 per 100,000 in 2000. A jointly agreed policy on MRSA and timely co-operation between all units were essential for control of this epidemic.