A validation of the Danish microbiology database (MiBa) and incidence rate of Actinotignum schaalii (Actinobaculum schaalii) bacteraemia in Denmark.

Actinotignum schaalii (former named Actinobaculum schaalii) can cause urinary tract infections (UTIs) and bacteraemia, mainly in the elderly. A. schaalii is difficult to identify with conventional biochemical tests, and it is often overlooked if the urine is only cultured in ambient air. The aim of this study was to validate data from the nationwide Danish microbiology database (MiBa) with data from the laboratory information system (LIS) at the local department of microbiology in Viborg-Herning, and to evaluate the incidence rate of bacteraemia caused by A. schaalii in Denmark by using data from the MiBa. All departments of microbiology in Denmark report data to the MiBa. All microbiological samples with A. schaalii in Denmark were extracted for a period of 5 years from the MiBa and from the local LISs. All data obtained from our local LIS were also found in the MiBa, except for data on real-time PCR, which were not registered, owing to missing ID codes in the MiBa. From 2010 to 2014, there was a significant increase in the incidence rate of blood cultures with A. schaalii, from 1.8 to 6.8 cases per million, which was probably due to coincident implementation of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) in routine diagnostics. We found that A. schaalii caused bacteraemia and UTIs mainly in the elderly. In conclusion, the MiBa can be a useful source of nationwide microbiological data in Denmark. Our results suggest that the incidence rate of A. schaalii as a cause of bacteraemia has been underestimated, and that culture of urine in CO2 can improve the detection of A. schaalii.

Fusobacterium necrophorum tonsillitis: an important cause of tonsillitis in adolescents and young adults.

The role of Fusobacterium necrophorum in tonsillitis in adolescents and young adults was retrospectively investigated by culture examination. We compared the prevalence of F. necrophorum in 212 subjects with confirmed clinical tonsillitis and in 176 subjects with confirmed no clinical tonsillitis. The prevalence of F. necrophorum was significantly higher (p < 0.001) in subjects with clinical tonsillitis (27%) compared to subjects with no clinical tonsillitis (6%). These results clearly demonstrate the role of F. necrophorum in tonsillitis. By diagnosing and treating F. necrophorum tonsillitis with, for example, penicillin, metronidazole, or both, we might prevent some cases of Lemierre syndrome.

A cost-effectiveness analysis of identifying Fusobacterium necrophorum in throat swabs followed by antibiotic treatment to reduce the incidence of Lemierre's syndrome and peritonsillar abscesses.

The main purpose of this paper was to estimate the cost per quality-adjusted life year (QALY) saved by identifying Fusobacterium necrophorum in throat swabs followed by proper antibiotic treatment, to reduce the incidence of Lemierre's syndrome and peritonsillar abscesses (PTA) originating from a pharyngitis. The second purpose was to estimate the population size required to indicate that antibiotic treatment has an effect. Data from publications and our laboratory were collected. Monte Carlo simulation and one-way sensitivity analysis were used to analyse cost-effectiveness. The cost-effectiveness analysis shows that examining throat swabs from 15- to 24-year-olds for F. necrophorum followed by antibiotic treatment will probably be less costly than most other life-saving medical interventions, with a median cost of US$8,795 per QALY saved. To indicate a reduced incidence of Lemierre's syndrome and PTA in Denmark, the intervention probably has to be followed for up to 5 years. Identifying F. necrophorum in throat swabs from 15- to 24-year-olds followed by proper antibiotic treatment only requires a reduction of 20-25 % in the incidence of Lemierre's syndrome and PTA to be cost-effective. This study warrants further examination of the effect of antibiotic treatment on the outcome of F. necrophorum acute and recurrent pharyngitis, as well as the effect on Lemierre's syndrome and PTA.

Rifampicin-soaked silver polyester versus expanded polytetrafluoro-ethylene grafts for in situ replacement of infected grafts in a porcine randomised controlled trial.

BACKGROUND: To compare the efficacy of in situ replacement with rifampicin-soaked silver-coated polyester (RSSCP) to that of expanded polytetrafluoroethylene (ePTFE) graft replacements in a porcine model for early aortic prosthetic vascular graft infection (PVGI). MATERIAL AND METHODS: Sixty pigs received polyester or silver-coated grafts with an 8 mm diameter implanted end-to-end in the infrarenal aorta, and the grafts were inoculated with approximately 10(6)Staphylococcus aureus. All developed S. aureus PVGI. Two weeks later, the 52 surviving pigs were randomised to undergo in situ graft replacement with ePTFE or RSSCP grafts followed by oral administration of 300 mg rifampicin and 750 mg ciprofloxacin twice a day, postoperatively. After three weeks, all pigs were sacrificed. In situ perigraft swabs and graft material were analysed for S. aureus quantitatively. RESULTS: Only one out of 25 RSSCP grafts were infected with S. aureus, whereas 15 of 27 ePTFE grafts were infected after 3 weeks (OR = 0.022, 95% CI: 0.002, 0.219, P = 0.001). CONCLUSIONS: In situ replacement with RSSCP grafts and oral rifampicin plus ciprofloxacin is more efficiency in eradicating S. aureus PVGI than ePTFE grafts treated with same oral antibiotics in a porcine aortic PVGI model.

Prevention of primary vascular graft infection with silver-coated polyester graft in a porcine model.

OBJECTIVES: To evaluate the efficacy of a silver-coated vascular polyester graft in the prevention of graft infection after inoculation with Staphylococcus aureus in a porcine model. MATERIAL AND METHODS: Eighty-four pigs were randomly selected 1:1 to receive a silver-coated or non-silver-coated 8-mm-wide polyester graft implanted end-to-end in the infrarenal aorta. At the end of implantation, 10(6) colony forming units (CFUs) S. aureus in 0.3ml suspension were inoculated directly on the graft surface. Blood samples assayed for white blood corpuscles (WBCs) and C-reactive protein (CRP) were taken before implantation and on the postoperative days 2, 5, 7, 11 and 14. Two weeks after implantation, the perigraft swabs were analysed for S. aureus or contaminants. CFUs of S. aureus were quantified and logarithmised. Student's t-tests, repeated measurement analysis of variance (ANOVA) and chi-square test were employed to compare the two grafts. RESULTS: All pigs developed graft infection. There were no statistically significant differences between the silver-coated and non-silver-coated grafts in the quantity of S. aureus, macroscopic signs of infection and postoperative changes in the temperature, WBC and CRP. CONCLUSIONS: Silver-coated polyester grafts failed to prevent graft infections after inoculation with 10(6)S. aureus in 0.3ml suspension in a porcine model.

Lemierre's syndrome and other disseminated Fusobacterium necrophorum infections in Denmark: a prospective epidemiological and clinical survey.

In a 3-year prospective study, all cases of disseminated Fusobacterium necrophorum infections found in Denmark from 1998 to 2001 were analysed, with the aim of describing the epidemiology and clinical features of the variants of Lemierre's syndrome and disseminated non-head-and-neck-associated F. necrophorum infections. Fifty-eight cases of Lemierre's syndrome were reported in previously healthy persons, with an incidence of 14.4 cases per million per year in youngsters aged 15-24 years old. There was no increase during the study period. Lemierre's syndrome originating from an oropharyngeal infection was seen in 37 youngsters. An otogenic variant of Lemierre's syndrome was seen in 5 children with dissemination to nearby regions, and other variants of Lemierre's syndrome, e.g. from the sinuses and teeth, were seen in 16 adults. Patients often had metastatic infections already on admission and needed prolonged hospitalisation. The overall mortality of Lemierre's syndrome was 9%. Forty-two elderly patients had disseminated F. necrophorum infections originating from foci in lower parts of the body. They frequently had predisposing diseases, e.g. abdominal or urogenital cancers, which contributed to the high mortality of 26%. This study shows that the incidence of Lemierre's syndrome is higher than that previously found and has a characteristic age distribution. Early suspicion of the diagnosis, several weeks of antibiotic therapy, often combined with surgical drainage, is mandatory to lower the mortality. In disseminated non-head-and-neck-associated F. necrophorum infections, underlying cancers must be considered.

Localised Fusobacterium necrophorum infections: a prospective laboratory-based Danish study.

During a 3-year prospective laboratory-based study in Denmark from 1998 to 2001, all patients who were diagnosed with localised Fusobacterium necrophorum infections were registered with the purpose of describing the variety of localised infections caused by F. necrophorum, especially in the head. We found 267 patients, most of them previously healthy, with localised F. necrophorum infections in the head and neck. In children, F. necrophorum caused otitis media and solitary abscess formation in cervical lymphadenitis. In adolescents, F. necrophorum was found in 21% of peritonsillar abscesses. We also found F. necrophorum in young adults with tonsillitis and in middle-aged adults with sinusitis. F. necrophorum was found in substantial amounts and as the only bacterial pathogen in the majority of patients. All 267 patients recovered without sequelae. We found another 21 localised non-head-and-neck-associated F. necrophorum infections, mainly subcutaneous wound infections in adults. This study shows that F. necrophorum causes a variety of localised infections, especially in the head and neck region, with a characteristic age distribution. We recommend that anaerobic culture is performed on swabs from children with recurrent otitis media and adolescents with non-streptococcal group A tonsillitis.

Minimum requirements for a rapid and reliable routine identification and antibiogram of Fusobacterium necrophorum.

Three hundred fifty-seven isolates of Fusobacterium necrophorum from human infections in Denmark were consecutively collected over a 3 year period for the purpose of establishing the minimum requirements for rapid and reliable routine identification of Fusobacterium necrophorum using phenotypic characters. The first 40 isolates were fully characterized by the most common phenotypic tests mentioned in the literature, while the last 317 where identified solely by the established minimum requirements for rapid and reliable routine identification of Fusobacterium necrophorum. All but one isolate were identical in all phenotypic tests. The outlying strain differed in morphology and the ability to agglutinate erythrocytes. On the basis of our findings it should be possible within 3-4 days to identify and differentiate F. necrophorum from other species including other Fusobacterium spp. by the unique but subspecies specific colony morphology, susceptibility to kanamycin and metronidazole, the smell of butyric acid, chartreuse colour fluorescence, and beta-haemolysis on horse blood agar. Three-hundred fifty-six isolates were identified as F. necrophorum subsp. funduliforme while one strain was F. necrophorum subsp. necrophorum. The species and subspecies level was confirmed for the first 40 isolates by real-time PCR. MIC in mg/l was determined for the 40 isolates. MIC(90) was 0.047 for penicillin, 0.047 for clindamycin, 0.25 for metronidazole, 0.38 for cefuroxime, >32 for imipenem, 0.012 for meropenem, and 2 for erythromycin. All 357 isolates were susceptible to penicillin and metronidazole indicating that these antibiotics are still the drugs of choice in antibiotic therapy of F. necrophorum infections, but therapy with clindamycin may be an alternative. Erythromycin should be avoided.

Laparoscopic diagnosis and treatment of aortic vascular prosthetic graft infections in a porcine model.

OBJECTIVES: To study the feasibility and efficacy of experimental laparoscopy in the diagnosis of aortic graft infection in pigs. MATERIAL AND METHODS: Eight pigs had an aortic tube graft implanted and inoculated with either 5 x 10(4) or 10(6) CFU of Staphylococcus aureus ATCC 29213. Laparoscopy was performed after a median of 20 days with debridement and sampling for bacterial culture. Thereafter, the grafts were locally soaked in rifampicin and postoperatively, the pigs received rifampicin and ciprofloxacin orally for two weeks and were then sacrificed. RESULTS: All pigs developed graft infection. One pig died from severe clostridial septicaemia before laparoscopy could be performed. The remaining pigs had all samples for bacterial culture taken by laparoscopy from the inflamed tissue. The temperature dropped significantly after laparoscopy, and no macroscopic signs of infection presented at autopsy. However, only culture from one pig was without S. aureus at autopsy. CONCLUSIONS: Laparoscopy is a potential diagnostic tool for aortic graft infection and also affords the opportunity to carry out bacteriological sampling and local antibiotic treatment. The efficacy of laparoscopic treatment needs further evaluation.

Detection of Fusobacterium necrophorum subsp. funduliforme in tonsillitis in young adults by real-time PCR.

Throat swabs from 61 patients, aged 18-32 years, with non-streptococcal tonsillitis (NST) and 92 healthy controls were examined for the presence of Fusobacterium necrophorum DNA using a novel TaqMan-based real-time quantitative PCR assay for F. necrophorum subspecies. The assay was based on the gyrB subunit gene, and detected F. necrophorum DNA in 48% of patients with NST and in 21% of controls (p <0.001). F. necrophorum subsp. funduliforme was the only subspecies found in both patients and controls. The load of F. necrophorum DNA on swabs from patients with NST was significantly higher than that on swabs from controls (p <0.001). Furthermore, patients with recurrent NST had a significantly higher load of F. necrophorum DNA compared to patients with acute NST (p 0.04). In addition, 26 patients with tonsillitis and group C streptococci (GCS) had a significantly higher load of F. necrophorum DNA compared to the NST group (p <0.001). It was concluded that F. necrophorum subsp. funduliforme is present in small numbers as part of the normal human throat flora, and that F. necrophorum in large quantities may cause tonsillitis, especially recurrent tonsillitis. In addition, the study suggests that the concomitant presence of GCS may aggravate F. necrophorum tonsillitis.

A five-year survey of pneumococcal peritonitis in two Danish counties--incidence, diagnosis and clinical entities.

Eighteen patients with pneumococcal peritonitis were hospitalized in a five-year survey period from a Danish catchment population of 730 000 (0.5/100 000/year). They were naturally divided into three groups: (1) six children with appendicitis; (2) eight women of childbearing age with pelvic inflammatory disease (PID); and (3) four elderly patients with ulcerating gastric diseases. Complications were observed in six patients. All infections were community-acquired, but the pneumococcal etiology was not suspected before the organism was isolated in the laboratory. A classification of peritonitis caused by Streptococcus pneumoniae is discussed.

Diagnosis of Chlamydia trachomatis infections in a sexually transmitted disease clinic: evaluation of a urine sample tested by enzyme immunoassay and polymerase chain reaction in comparison with a cervical and/or a urethral swab tested by culture and polymerase chain reaction.

OBJECTIVE: To evaluate the value of a urine sample for diagnosing Chlamydia trachomatis infection in an STD clinic in a prospective study of samples collected from 410 consecutive STD patients (167 female and 243 male). METHODS: Urine samples were tested by enzyme immunoassay (EIA) and polymerase chain reaction (PCR) in comparison with cervical and/or urethral swabs tested by PCR and cell culture. A questionnaire was completed for a total of 320 patients concerning symptoms, and determining whether they were controls, contacts or were being tested subsequent to legal abortion. RESULTS: The overall prevalence of C. trachomatis infection was 11.5%. At least 40% of patients were asymptomatic. Of the C. trachomatis-positive patients, 85% were diagnosed by testing urine, compared to 91% by testing swabs. For urine tests, the sensitivities of PCR were 66.7% and 71.9% for female and male patients, respectively, and the sensitivities of EIA were 40.0% and 62.5%, or 46.7% and 71.9%, for female and male patients, respectively, by including a 30% gray zone below the cut-off value. For swabs, the sensitivities of PCR were 93.3% and 87.5% for female and male patients, respectively, and equal to the sensitivities of culture. In total, 3.3% of controls and 35% of contacts were found to be C. trachomatis positive. CONCLUSION: The use of urine samples for the diagnosis of C. trachomatis infections was effective, but urine samples should be additional to conventional swab(s) instead of replacing. Partner notification and a confirmation of cure is recommended.

A 7-month outbreak of relapsing postpartum group A streptococcal infections linked to a nurse with atopic dermatitis.

A 7-month outbreak of 15 cases of postpartum sepsis with group A haemolytic Streptococci (GAS) was stopped when a carrier was identified. Comparing delivery dates with duty rotas revealed that the carrier had been present during delivery in 13 of the 15 cases. The epidemic GAS type, T3-13-B3264, was found in a carbuncle in her groin and in atopic dermatitis lesions behind her ears and on her eyelids. Thus, it was not the microbiological screening of staff that helped detect the carrier. The outbreak went unnoticed for 6 months, as no 2 cases were diagnosed by the same physician and 5 cases were diagnosed by different general practitioners. The main risk factors for infection were presence of the carrier relative risk (relative risk RR 47.8, 95% confidence interval (CI) 10.9-209.5) and suturing of episiotomy (RR 11.0; 95% CI 2.6-47.9). We recommend that a thorough epidemiological investigation should be carried out in every single case of GAS postpartum infection. Despite initial intravenous treatment with penicillin, 8 patients experienced > 15 recurring postpartum GAS infections, such as endometritis, wound infection, tonsillitis, erysipelas and Brodie's abscess. Eradication of GAS should be confirmed after completion of treatment.

Isolation of Kingella kingae from synovial fluids using four commercial blood culture bottles.

According to the literature, Kingella kingae may be an underdiagnosed cause of joint and bone infections in children. The use of the Bactec blood culture system for culture of joint fluids has dramatically improved the isolation of this fastidious bacterium. The aim of this study was to test the recovery rate and detection time of four commercial blood culture systems: three different BacT/Alert (Organon Teknika, USA) bottles and one Bactec (Becton Dickinson Microbiology Systems, USA) bottle, all inoculated with Kingella kingae strains mixed with pooled synovial fluids. For each strain the same inoculum and volume of synovial fluid was distributed into each of the four bottles. All 24 strains tested grew in the BacT/Alert Aerobic (100%) and the BacT/Alert Pedi-BacT (100%) bottles. Twenty-one strains grew in the BacT/Alert FAN aerobic (88%) bottle, and 15 strains grew in the Bactec Plus Aerobic F (63%) bottle, in both systems within 12 days (P<0.01). The Kingella kingae strains were first detected in the BacT/Alert Pedi-BacT bottles (P<0.001). The results were reproducible. The BacT/Alert blood culture bottles were superior to previously described blood culture systems in isolating Kingella kingae from synovial fluid, even with small inoculums and small volumes of synovial fluid.

Human necrobacillosis, with emphasis on Lemierre's syndrome.

Lemierre's syndrome is the classical presentation of human necrobacillosis. It is characterized by a primary infection in the head in a young, previously healthy person who subsequently develops persistent high fever and disseminated metastatic abscesses, frequently including a septic thrombophlebitis of the internal jugular vein. The main pathogen is Fusobacterium necrophorum, an obligate anaerobic, pleomorphic, gram-negative rod. Clinical microbiologists have a key role in alerting clinicians and advising proper antibiotic treatment when the characteristic microscopic morphology of the pleomorphic F. necrophorum is seen in Gram stains from positive anaerobic cultures of blood and pus. Early diagnosis and prolonged appropriate antibiotic treatment with good anaerobic coverage are crucial to reduce morbidity and mortality. F. necrophorum also causes human necrobacillosis with foci caudal to the head, mainly in elderly patients with high mortality related to age and predisposing diseases, such as cancers of the primary focus.

Survival and function of phagocytes in blood culture media.

The survival and function of human phagocytes in sterile aerobic and anaerobic blood culture media were investigated using neutrophil morphology, white blood cell count in a haemoanalyser, flow cytometry, oxidative burst response, and bactericidal effect in Colorbact and Septi-Chek blood culture media and Bact/Alert. When comparing agitation to stationary incubation no difference in phagocytic activity was found. The methods showed the same trends demonstrating that the phagocytes' viability and activity were prolonged by oxygen and shortened by anaerobic conditions and sodium polyethanol sulfonate (SPS). Best preserved activity and viability were found in the aerobic media containing less than 0.5 g/l SPS, in which significant phagocyte oxidative burst and bactericidal activity were found up to 4 days after inoculation. Considering that the majority of bacteremias are due to aerobic or facultatively anaerobic bacteria, the present data suggest that most bacteria may be recovered by the use of one aerobic bottle with SPS concentration below 0.5 g/l to protect meningococci and other SPS-sensitive bacteria and one above 0.5 g/l to stop phagocytic activity, plus one anaerobic bottle with SPS below 0.5 g/l.