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.

To establish the incidence and describe the clinical epidemiology of necrobacillosis and Lemierre's syndrome in Denmark, the clinical records of all laboratory-recorded cases of septicaemia due to Fusobacterium necrophorum biovar A, B, and C were reviewed retrospectively during a 6-year period. The incidence of necrobacillosis and Lemierre's syndrome was 1.5 and 0.8 per million persons per year, respectively, showing a tendency to increase during the period. Fusobacterium necrophorum was grown after three days' incubation, but the characteristic pleomorphic fusiform morphology was often disregarded as an important help in diagnosing necrobacillosis. The 24 patients with Lemierre's syndrome were all young and previously healthy, and none died, but pre-hospital delay was associated with a significantly higher morbidity and risk of metastatic infections. The remaining 25 patients with necrobacillosis had a high mortality, 24%, which was correlated with age and predisposing diseases, especially cancers. These findings stress the importance of a quicker clinical and microbiological diagnosis in cases of Lemierre's syndrome, and of screening for cancer in the remaining cases of necrobacillosis.

Staphylococcus lugdunensis, a rare cause of severe infections such as native valve endocarditis, often causes superficial skin infections similar to Staphylococcus aureus infections. We initiated a study to optimize the identification methods in the routine laboratory, followed by a population-based epidemiologic analysis of patients infected with S. lugdunensis in Viborg County, Denmark. Recognition of a characteristic Eikenella corrodens-like odor on Columbia sheep blood agar combined with colony pleomorphism and prominent ␤-hemolysis after 2 days of incubation, confirmed by API-ID-32 Staph, led to an 11-fold increase in the detection of S. lugdunensis. By these methods we found 491 S. lugdunensis infections in 4 years, corresponding to an incidence of 53 per 100,000 per year, an increase from 5 infections per 100,000 inhabitants in the preceding years. Seventy-five percent of the cases were found in general practice; these were dominated by skin abscesses (36%), wound infections (25%), and paronychias (13%). Fifty-six percent of the infections occurred below the waist, and toes were the most frequently infected site (21%). Only 3% of the patients suffered from severe invasive infections. The median age was 52 years, and the male/female ratio was 0.69. Our study shows that S. lugdunensis is a common cause of skin and soft-tissue infections (SSTI) and is probably underrated by many laboratories. S. lugdunensis should be accepted as a significant pathogen in SSTI and should be looked for in all routine bacteriological examinations, and clinicians should be acquainted with the name and the pathology of the bacterium.Staphylococcus lugdunensis, one of the coagulase-negative staphylococci (CoNS), was first described by Freney et al. 20 years ago (8). It causes severe infections similar to Staphylococcus aureus infections, especially acute endocarditis in prosthetic and native valves (14,24,29,30,36). Other infections such as osteomyelitis (17, 23), peritonitis (28), intravascular catheter infections (5, 32), prosthetic joint infections (26), and urinary tract infections have also been reported (10). S. lugdunensis has been reported as an important cause also of skin and soft tissue infections (SSTI), primarily in the groin (1, 39) and mammae (13,22,25,37,41).S. lugdunensis is part of the normal human skin flora and commonly colonizes the perineal region (1, 37, 39); however, carriage rates of different CoNS are not comprehensively accounted for, and only a few have looked for S. lugdunensis carriage (13, 16, 39).Our study was undertaken after two serious cases of S. lugdunensis infection, a fatal case of endocarditis with protracted bacterial identification but correct antibiotic treatment and a case of late-diagnosed osteomyelitis, which prompted an optimization of our routine microbiological methods for identification of S. lugdunensis. This subsequently increased the number of S. lugdunensis isolates significantly, especially in samples from patients in general practice, and encouraged us to conduct a prospectiv...

A preliminary epidemiological survey indicated an association between Ascaris infections in Danish patients and contact with pigs or pig manure. In the present study, we compared Ascaris worms collected from humans and Ascaris worms collected from pigs by amplified fragment length polymorphism (AFLP) analysis, a technique for whole-genome fingerprinting, and by PCR-linked restricted fragment length polymorphism (PCR-RFLP) analysis of the internal transcribed spacer region of nuclear rDNA. The AFLP data were analyzed by distance-and model-based clustering methods. These results assigned Ascaris worms from Danish patients to a cluster different from that for worms from humans in other geographic areas. In contrast, worms from humans and pigs in Denmark were assigned to the same cluster. These results were supported by the PCR-RFLP results. Thus, all of the examined Danish patients had acquired Ascaris infections from domestic pigs; ascariasis may therefore be considered a zoonotic disease in Denmark.

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.

Nine of 10 strains of Actinobaculum schaalii caused urinary tract infections in predisposed individuals. Identification included 16S rRNA gene sequence analysis and use of the API Coryne and Rapid ID32A test systems. A. schaalii is easily overlooked due to its slow growth in ambient air and its resemblance to the normal bacterial flora on skin and mucosa.The genus Actinobaculum, first described in 1997, includes the Actinobaculum suis and Actinobaculum schaalii species. A. suis is an important cause of urinary tract infections (UTIs) and abortions in sows and was formerly assigned to a variety of genera, including Corynebacterium, Eubacterium, and Actinomyces (9, 17). A. schaalii is a new species recovered from human blood and urine and is suspected to cause UTIs (9, 12). Two newly described species, Actinobaculum massiliae and Actinobaculum urinale, were recovered from the urine of elderly women with chronic cystitis (6, 7). A. massiliae is also described as a new cause of superficial skin infections (16). Problems with identifying Actinobaculum spp. with traditional phenotypic tests have obscured their pathological role for many years. We describe 10 cases of A. schaalii infections.Nine strains of A. schaalii were available for growth, biochemical, and susceptibility tests. The strains were cultured on nine different culture media at 35°C in ambient air, in air with 5% CO 2 , and anaerobically. The media were 5% Columbia sheep blood agar (Becton Dickinson [BD], Heidelberg, Germany), 5% and 10% horse blood agar (Statens Serum Institut [SSI], Copenhagen, Denmark), chocolate agar (SSI), Brucella blood agar with hemin and vitamin K1 (BD), anaerobic plates (SSI), nutrient agar plates (SSI), semisolid agar containing pepsin blood and thioglycolate (SSI), and serum broth (SSI). The CAMP reaction was performed on CAMP plates containing sheep erythrocytes (SSI) with a streak of a beta-hemolytic strain of Staphylococcus aureus. The strains were characterized by using the API Coryne and Rapid ID32A systems in accordance with the manufacturer's instructions (API bioMérieux, Marcy l'Etoile, France). Carbohydrate fermentation reactions were read after 24 and 48 h of incubation.A Quantitect SYBR green kit (QIAGEN) was used with real-time PCR mixtures (50-l total volume) containing 1ϫ PCR buffer and a 200 M concentration of each primer. The primers used for amplification of the 16S rRNA gene, BSF-8 (5Ј-AGAGTTTGATCCTGGCTCAG-3Ј) and BSR-534 (5Ј-ATTACCGCGGCTGCTGGC-3Ј), produced a 526-bp fragment of the 16S rRNA gene. Samples of 1 and 5 l were tested by PCR. PCRs were performed by using an Opticon DNA engine (MJ Research). The amplification profile included incubation at 95°C for 15 min followed by 40 cycles at 94°C for 30 s, 55°C for 30 s, and 72°C for 30 s. PCR samples were spin column purified using Microcon YM-100 filter units (Millipore) for DNA sequencing. DNA strands of the amplicons were sequenced on an ABI PRISM 3100 Avant genetic analyzer (Applied Biosystems) using BSF-8 and BSR-534 as sequencing primers and a BigDye v....

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.

This organism is identified more often by PCR than by cultivation.