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The authors have observed a case of Fusobacterium necrophorum associate with Actinomyces odontolyticus septicemia, which appears to be the first case reported. In a 19 year old man, 15 blood cultures allowed to isolate both germs. Clinical manifestations were: acute-like abdomen, encephalitis, myocarditis and pleural effusion. Treatment with amoxicillin and metronidazole was successful.
Fusobacterium necrophorum septicemia developed in five patients after an oropharyngeal infection. Four patients had sore throat or neck pain, and two had findings of jugular vein septic thrombophlebitis. Metastatic abscesses, including embolic pneumonia, empyema, septic arthritis, and osteomyelitis, also occurred. Four patients recovered and one died. Proper treatment requires recognition of the oropharyngeal source of the septicemia and its differentiation from endocarditis. Antibiotic therapy should be prolonged, and metastatic abscesses drained.
Isolates of Fusobacterium necrophorum were obtained from 124 bovine hepatic abscesses (in 119 cattle) and from the ruminal contents of 12 cattle. Three biotypes (A, AB, and B) were found. Type A isolates were most common in hepatic abscesses, and type B isolates were most common in ruminal contents. Virulence studies in mice indicated that type A isolates were more pathogenic than types AB or B isolates. The difference in virulences between types A and B isolates was not related to the toxicity of the endotoxin. Results of serotesting (passive hemagglutination) showed that a wide heterogeneity existed between isolates. Minimal inhibitory concentrations of antibiotics for 12 of the isolates (7 type A, 1 type AB, and 4 type B) were as follows: less than or equal to 0.06 IU of penicillin G/ml; less than or equal to 0.5 micrograms of cephaloridine/ml; less than or equal to 2 micrograms of chloramphenicol/ml; less than or equal to 0.06 micrograms of clindamycin/ml; less than or equal to 8 micrograms of erythromycin/ml; less than or equal to 128 micrograms of gentamicin/ml; less than or equal to 0.25 micrograms of oxytetracycline/ml; and less than or equal to 4 micrograms of tylosin/ml.
Spondylodiscitis is rarely caused by anaerobic germs and occurs mainly in patients weakened by another infection. We report a case of Fusobacterium nucleatum spondylodiscitis in a 63-year-old man. This is the third such report and the second in a non-immunodepressed patient. The clinical presentation and laboratory findings in this type of spondylodiscitis vary little from those produced by other germs. Magnetic resonance imaging is the most useful diagnostic tool for spondylodiscitis. The germ is isolated from a discal puncture aspirate in 66% of the cases but can also be obtained from blood cultures. Anaerobic germs are usually sensitive to penicillin or cephalosporins.
A 26-year-old man was admitted to our hospital with a 10-day history of sore throat, high fever, and right knee joint pain. On physical examination, the pharynx was considerably inflamed, and the right knee joint was swollen and extremely tender. Chest radiography showed multiple, bilateral nodules and masses with pleural effusions. Fusobacterium necorforum grew from samples of blood, pleural effusion, and pus taken from the knee joint. The patient was treated with intravenous clindamycin, ventilatory support, and continuous chest and knee joint drainage. His condition progressively improved and he was discharged on the 66th hospital day. A disease caused by an oropharyngeal infection with secondary suppurative thrombophlebitis of the internal jugular vein, and complicated by multiple metastatic infections is called postanginal septicemia, or Lemierre syndrome. Before the discovery of antibiotics, this disease usually was fatal. The widespread use of antibiotics for treat oropharyngeal infections may have caused a number of reported cases. Lemierre syndrome is an uncommon complication of oropharyngeal infection, and it may be fatal if diagnosis is delayed. Careful attention must be directed to patients with oropharyngeal infection who have signs and symptoms that suggest metastatic infection.
A case of multiple liver abscesses caused by Fusobacterium necrophorum infection is presented. In cases with fever of unknown origin and biochemical signs of derangement of hepatic function, it is recommended that an abdominal scan is performed. Before initiating antibiotic treatment relevant specimens should be collected. Finally we describe how the use of resin-containing blood culture medium enhance the possibility of bacterial detection in patients receiving antibiotics.
A patient with pulmonary Fusobacterium nucleatum infection presenting as a solitary nodule is reported. The infectious nature of the lung nodule was disclosed by ultrasound-guided percutaneous transthoracic fine needle aspiration cytology (FNAC) with Liu's stain. The cytologic characteristics of F nucleatum infection are described. The usefulness of real-time ultrasound in fine needle aspiration diagnosis of a peripheral lung lesion is demonstrated, and the role of Liu's stain in FNAC is emphasized.
Postanginal sepsis or Lemierre's syndrome is characterised by septic thrombophlebitis of the jugular vein, metastatic abscesses in the lungs, soft tissues, joints or elsewhere, occurring several days to two weeks after tonsillitis or pharyngitis. The primary pathogen is a Gram-negative anaerobic rod, mostly Fusobacterium necrophorum. Previously healthy, young adults are affected mainly and the syndrome was seen more frequently in the pre-antibiotic era than it is nowadays. In the three young patients described here, a girl aged 15 and two boys aged 18 and 16, F. necrophorum was isolated from blood or pus. Histories and examinations were suggestive of Lemierre's syndrome. Ultrasound and CT scanning of the neck and other localisations proved to be important diagnostic tools in assessing the diagnosis. Response to therapy was slow and depended in at least one case on adequate drainage of abscesses. If the syndrome is suspected, initial antibiotic treatment should provide adequate coverage of anaerobic bacteria. In previously healthy patients with chills and fever occurring several days after a sore throat, Lemierre's syndrome should be considered.
OBJECTIVE: To determine the efficacy of leukotoxin-based Fusobacterium necrophorum vaccines and dietary tylosin in providing protection against experimentally induced hepatic abscesses in steers. DESIGN: 30 steers assigned randomly to 6 treatment groups of 5 steers each: 1, phosphate-buffered saline solution (PBSS; control); 2, PBSS control, fed tylosin (100 mg/steer) daily; 3, inactivated whole-cell culture with oil emulsion adjuvant; 4, culture supernatant (crude toxoid) with oil emulsion adjuvant; 5, semipurified leukotoxoid with oil emulsion adjuvant; and 6, semipurified leukotoxoid with saponin adjuvant. PROCEDURE: Steers were inoculated SC with emulsified antigen or PBSS on days 0 and 21. Blood samples were collected at weekly intervals to monitor serum antileukotoxin antibody titer. On day 42, all steers were challenge exposed intraportally with F necrophorum culture. Three weeks later (day 63), steers were euthanatized and necropsied to examine liver and assess protection. RESULTS: Antileukotoxin antibody titers of all vaccinated groups markedly increased from baseline values, and mean titers of vaccinated groups were higher than those of the control and tylosin-treated groups. Steers vaccinated with culture supernatant with oil emulsion adjuvant or semipurified leukotoxoid with saponin adjuvant had the highest mean antibody titers. All 5 steers in the control group developed liver abscesses. Tylosin feeding did not protect steers challenge exposed with F necrophorum intraportally. CONCLUSIONS: Culture supernatant was more protective than whole-cell culture or semipurified leukotoxin against experimentally induced hepatic abscesses. Partial purification of leukotoxin appeared to reduce its protective immunity.
An enzyme-linked-immunosorbent assay (ELISA) with HCl heat-extracted antigen of Fusobacterium necrophorum was conducted to detect specific immunoglobulins G and M in infected cattle. The ELISA revealed an increase (> 0.40) in specific IgG in most of the animals with hepatic abscesses but not that in specific IgM. All the lesions were positive for F. necrophorum. These findings indicated that the ELISA for immunoglobulin G detection may prove to be a useful tool for predictive serodiagnosis of F. necrophorum infection in cattle.
We report a case of spinal epidural abscess. In our knowledge this is the first ever reported case due to Fusobacterium nucleatum. An 83 year old female patient with a history of lumbar trauma presented severe lumbalgia. Magnetic resonance showed the unusual anterior location, complicated by psoas abscess. Laminectomy and lumbar drainage were performed. She was treated with ceftriaxone plus metronidazole during 10 days. The evolution was favourable without any residual neurological morbidity.
An unusual case of necrotic infection of the tongue and unilateral necrotic maxillar sinusitis, in a cow is reported. The prominent clinical findings were: a marked salivation, unilateral mucopurulent nasal discharge, diffuse necrotic lesions on the ventral side of the tongue and fetid breath. Clinico-pathological and pathological findings and abnormalities in the haemogram were probably caused by the toxins produced by the causative organism--Fusobacterium necrophorum like microorganism.
In the two cases of anaerobic meningitis and bacteremia caused by Fusobacterium species described, upper respiratory tract infection preceded the onset of meningitis. Isolation of the causative organism and appropriate antibiotic therapy resulted in cures in both patients. Although extremely rare, these organisms should be considered as possible causative agents in meningitis when routine cultures are negative, and appropriate anaerobic culture techniques should be employed, especially when sinus, otitic, or mastoid symptoms precede or accompany the onset of meningitis. Susceptibility testing of these organisms is necessary to ensure proper therapy because of the resistance of some anaerobes to antibiotics commonly employed in meningitis. In this regard, metronidazole, which provides excellent cerebrospinal fluid levels, may prove to be an effective agent in anaerobic infections of the central nervous system.
DNA from six strains of Fusobacterium nucleatum was examined for possible methylation of adenine or cytosine in the restriction site GATC. F. nucleatum DNA was digested with the restriction enzymes Sau3AI, DpnI and MboI, subjected to electrophoresis in agarose gels and Southern blotting. The blots were hybridized with a polymerase chain reaction (PCR)-amplified DNA probe. Both radioactive and non-radioactive labeling methods were used. Some strains of F. nucleatum were found to be dam methylated or dcm methylated in the sequence GATC; the adenine residue is methylated in strain ATCC 10953, while the cytosine residue is methylated in the strains F1 and F3. Strains F6, ATCC 25586 and Fev1 are probably not methylated in this nucleotide sequence.
The major outer membrane protein, FomA, of the Gram-negative human oral pathogen Fusobacterium nucleatum functions as a porin and is assumed to act as a receptor protein in coaggregation with other oral pathogenic bacteria such as Streptococcus sanguis and Porphyromonas gingivalis. We describe here the cloning of fomA from F. nucleatum in E. coli. Using pGEM3Zf(+), three recombinant plasmids were carrying parts of the fomA gene, but none of these contained regions upstream of the coding sequence. From these plasmids a clone was constructed which contained the whole fomA gene. The ATCC 10953 fomA gene was cloned under the phosphate limitation-inducible phoE promoter, using a vector derived from pACYC184. The protein was found to be incorporated into the outer membrane of the host in an apparently normal manner, as judged by heat-modifiability, trypsin-accessibility, and accessibility to antibodies to the protein in a whole cell enzyme-linked immunosorbent assay. The cloned FomA was found to exhibit pore-forming activity.
The gram-negative anaerobic bacterium, Fusobacterium nucleatum, is a predominant member of the human oral flora. As a major component of subgingival plaque, this bacterium has a significant impact on the ecology of the oral cavity due to its ability to adhere to many different microbial species. The objective of this study was to identify and characterize plasmids and transposons that may have the potential to be developed into tools for cloning, genetic transformation, and mutagenesis of oral isolates of F. nucleatum. Analysis of a collection of laboratory strains resulted in the identification of a homologous family of small cryptic plasmids. Plasmids within this family ranged in size from 6.0 to 6.6 kb. Eighteen percent of all strains examined (n = 74) contained DNA sequences related to the plasmids. Homologous plasmid sequences were found in strains belonging to 2 of the 3 subspecies of the bacterium. The 2 smallest plasmid species were cloned in Escherichia coli to facilitate endonuclease restriction mapping. Among the strains examined for plasmids, 5 exhibited resistance to at least 10 micrograms/ml of tetracycline. These strains, all members of the subsp. polymorphum, contained a tetracycline resistance determinant (TetM) as part of a Tn916-like integrated transposon sequence. The Tn916-like element and 1 of the plasmid species co-resided in a single strain of the bacterium. Hybridization patterns of the Tn916-like sequences were identical in all 5 tetracycline-resistant strains. However, these strains appeared to be clonally distinct based on genomic fingerprinting.
Anaerobically prepared cell-free extracts from Fusobacterium nucleatum contain 2-hydroxyglutaryl-CoA dehydratase with a specific activity of 20 nkat mg-1. The enzyme was purified 24-fold to a specific activity of 480 nkat mg-1 by anion exchange chromatography, gel filtration and chromatography on Blue-Sepharose. The activity of the purified enzyme was strictly dependent on the reductant Ti(III)citrate and stimulated 25-fold by 0.15 mM ATP and 5 mM MgCl2. ATP is hydrolysed to ADP during incubation with 2-hydroxyglutaryl-CoA dehydratase in the presence or absence of the substrate. The enzyme is extremely sensitive towards oxygen and is inhibited by 10 microM chloramphenicol, 10 microM 2,4-dinitrophenol or 0.15 mM hydroxylamine. The pure enzyme consists of three subunits alpha (49 kDa), beta (39 kDa) and gamma (24 kDa) in approximately equal amounts. In this respect the enzyme differs from the related 2-hydroxyglutaryl-CoA dehydratase from Acidaminococcus fermentans and lactyl-CoA dehydratase from Clostridium propionicum both of which are composed of only two subunits with sizes comparable to those of alpha and beta but require an additional protein for activity. The relative molecular mass of the native enzyme of about 100 kDa suggests a trimeric alpha beta gamma-structure. The homogeneous enzyme contains riboflavin (0.5 mol/112 kDa), iron and sulfur (3.5 mol/112 kDa each). Polyclonal antibodies directed against the 2-hydroxyglutaryl-CoA dehydratase from A. fermentans did not crossreact with cell free extracts or purified dehydratase from F. nucleatum. A comparison of the N-terminal amino acid sequences of the dehydratase subunits from A. fermentans and F. nucleatum, however, showed some similarities in the beta-subunits.
Membrane preparations of Fusobacterium nucleatum grown on glutamate contain glutaconyl-CoA decarboxylase at a high specific activity (13.8 nkat/mg protein). The enzyme was solubilized with 2% Triton X-100 in 0.5 M NaCl and purified 63-fold to a specific activity of 870 nkat/mg by affinity chromatography on monomeric avidin-Sepharose. The activity of the decarboxylase was strictly dependent on Na+ (Km = 3 mM) and was stimulated up to 3-fold by phospholipids. The glutaconyl-CoA decarboxylases from the gram-positive bacteria Acidaminococcus fermentans and Clostridium symbiosum have a lower apparent Km for Na+ (1 mM) and were not stimulated by phospholipids. In addition only the fusobacterial decarboxylase required sodium ion for stability and was inactivated by potassium ion. By incorporation of this purified enzyme into phospholipids an electrogenic sodium ion pump was reconstituted. The enzyme consists of four subunits, alpha (m = 65 kDa), beta (33 kDa), gamma (19 kDa), and delta (16 kDa) with the functions of a carboxy transferase (alpha), a carboxy lyase (beta and probably delta) and a biotin carrier (gamma). The subunits are very similar to those of the glutaconyl-CoA decarboxylases from the gram-positive bacteria. With an antiserum directed against the decarboxylase from A. fermentans the alpha- and the biotin containing subunits of the three decarboxylases and that from Peptostreptococcus asaccharolyticus could be detected on Western blots.