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The adverse effect of dilution on the infectivity of Fusobacterium necrophorum culture.

Dilution had an adverse effect on the infectivity of 24 h cultures of a strain of Fusobacterium necrophorum, which became apparent at or near the minimum lethal dose. Thus in mice inoculated subcutaneously the mortality produced by 0.01 ml of undiluted culture was almost invariably greater than that produced by 0.1 ml of a 1 in 10 dilution. The explanation appeared to lie in the increased physical separation of bacterial cells that was the inevitable consequence of dilution.

Animals↗

Further observations on enhancement of the infectivity of Fusobacterium necrophorum by other bacteria.

It had already been shown with a single virulent strain (A42) of Fusobacterium necrophorum that suspension of the fusobacteria in sub-lethal doses of broth cultures of other bacteria reduced the minimum infective dose (greater than 10(6) organisms) for mice by subcutaneous inoculation, sometimes to less than 10 organisms. The present study extended the known range of bacteria with strong infectivity-enhancing properties to include Bacillus cereus, Klebsiella oxytoca and Staphylococcus aureus; and those with weaker effect to include Bacillus subtilis, 'Bacteroides melaninogenicus', Clostridium sporogenes, Pasteurella haemolytica, and Proteus mirabilis. The study also showed that five further virulent strains of F. necrophorum closely resembled A42 in respect of striking susceptibility to infectivity enhancement by Escherichia coli. Actinomyces (Corynebacterium) pyogenes and S. aureus. One further strain (A6) of F. necrophorum resembled A42 in respect of strong infectivity enhancement by A. pyogenes, S. aureus, B. cereus and K. oxytoca but differed from it and the other five strains in being only slightly affected by E. coli. This work was a necessary prelude to the development of a method, based on infectivity enhancement, for the detection and isolation of F. necrophorum present in small numbers in heavily contaminated material such as faeces.

Actinomycosis↗

A sensitive method for isolating Fusobacterium necrophorum from faeces.

The isolation of Fusobacterium necrophorum present in small numbers in heavily contaminated material such as faeces or soil is hampered by the lack of an efficient selective medium and by the high minimum infective dose of the organism. A sensitive method for the detection and isolation of faecal strains of F. necrophorum type A was based on the subcutaneous injection of faeces, suspended (5% w/v) in broth culture of Actinomyces (Corynebacterium) pyogenes or Staphylococcus aureus to increase fusobacterial infectivity, into mice pretreated with clostridial antitoxins. When necrobacillosis developed F. necrophorum was identified microscopically in tissue from the advancing edge of the lesion and isolated on a partly selective medium. The enhancement of fusobacterial infectivity produced by A. pyogenes and by S. aureus was high, but the latter was slightly the more efficient, enabling as few as 80 F. necrophorum organisms/g of faeces to be detected. Use of the method showed that 3 of 16 wallabies had F. necrophorum in their faeces at the time of examination. Numerous epidemiological applications are suggested.

Animals↗

Pathogenicity of Fusobacterium necrophorum strains from man and animals.

Necrobacillosis occurs in man and animals. The typical forms of the disease in animals are caused by Fusobacterium necrophorum biovar A; biovar B strains are much less pathogenic. In this study the pathogenicity for mice of eight human isolates of F. necrophorum was compared with that of animal biovar A and B strains. By subcutaneous inoculation seven of the human strains differed from biovar A but resembled biovar B in (1) producing, at the most, mild local lesions that rapidly healed, and (2) showing no enhancement of infectivity when suspended in sub-lethal doses of Staphylococcus aureus broth culture. The eighth human strain (A2433) resembled biovar A but differed from biovar B in (1) producing severe lesions, and (2) showing greatly enhanced infectivity in the presence of S. aureus. Nonetheless, strain A2433 differed from biovar A, both in the nature of the lesions produced and in its failure to cause severe general signs of illness and rapidly fatal infection. By intravenous inoculation one of two biovar B strains and all except one of the eight human strains produced purulent lesions, often severe, in the liver and elsewhere, but infection was not usually associated with general signs of illness. In contrast, intravenous injection of a biovar A strain gave rise to a rapidly fatal infection, with severe lesions in the liver or elsewhere. The results suggest that the term 'necrobacillosis' as used in human and veterinary medicine refers to diseases that differ in important respects.

Animals↗

Effect of disturbance of the gastrointestinal microflora on the faecal excretion of Fusobacterium necrophorum biovar A.

Oral pretreatment of mice with either a mixture of kanamycin and erythromycin or metronidazole to modify the gut microflora greatly enhanced the faecal excretion of Fusobacterium necrophorum biovar A given by mouth. This lends support to the suggestion that disturbance of the gastrointestinal microflora in animals such as cattle, which often carry the organism in the rumen, may lead to intestinal multiplication and faecal excretion, thereby providing a source of infection that may lead to necrobacillosis of the body surface.

Animals↗

Clonality of Fusobacterium nucleatum in root canal infections.

Fusobacterium nucleatum is a gram-negative non-spore-forming, non-motile, obligate anaerobic rod that is normally isolated from the oral cavity. Several studies have reported a significant heterogeneity within the F. nucleatum species. The aim of the present study was to analyze the clonal diversity of F. nucleatum strains isolated from intracanal infections and to evaluate the presence of Enterobacterial Repetitive Intergenic Consensus (ERIC)-like sequences in the genome of F. nucleatum. Samples were collected from 13 single-root teeth from adult patients, all having carious lesions, necrotic pulps and radiographic evidence of periradicular bone loss. F. nucleatum was isolated from two different patients (subjects 5 and 7) by culture. Amplification of 19 colonies from subject 5 and 15 colonies from subject 7 using ERIC primers resulted in four clonal types, two per subject. An intense amplicon of approximately 700 bp was generated by ERIC-PCR for all F. nucleatum isolates and F. nucleatum ssp. polymorphum ATCC 10953. The amplification reaction using primer 1254 confirmed the results obtained with the ERIC primer. Our findings indicate that DNA fingerprints provided by ERIC- and Arbitrarily Primed (AP)-PCR may constitute a powerful tool for investigating F. nucleatum clonal diversity.

Adult↗

Effect of Fusobacterium nucleatum on the T and B cell responses to Porphyromonas gingivalis in a mouse model.

T cell cytokine profiles and specific serum antibody levels in five groups of BALB/c mice immunized with saline alone, viable Fusobacterium nucleatum ATCC 25586, viable Porphyromonas gingivalis ATCC 33277, F. nucleatum followed by P. gingivalis and P. gingivalis followed by F. nucleatum were determined. Splenic CD4 and CD8 cells were examined for intracytoplasmic interleukin (IL)-4, interferon (IFN)-gamma and IL-10 by dual colour flow cytometry and the levels of serum anti-F. nucleatum and anti-P. gingivalis antibodies determined by an ELISA. Both Th1 and Th2 responses were demonstrated by all groups, and while there were slightly lower percentages of cytokine positive T cells in mice injected with F. nucleatum alone compared with the other groups immunized with bacteria, F. nucleatum had no effect on the T cell production of cytokines induced by P. gingivalis in the two groups immunized with both organisms. However, the percentages of cytokine positive CD8 cells were generally significantly higher than those of the CD4 cells. Mice immunized with F. nucleatum alone had high levels of serum anti-F. nucleatum antibodies with very low levels of P. gingivalis antibodies, whereas mice injected with P. gingivalis alone produced anti-P. gingivalis antibodies predominantly. Although the levels of anti-F. nucleatum antibodies in mice injected with F. nucleatum followed by P. gingivalis were the same as in mice immunized with F. nucleatum alone, antibody levels to P. gingivalis were very low. In contrast, mice injected with P. gingivalis followed by F. nucleatum produced equal levels of both anti-P. gingivalis and anti-F. nucleatum antibodies, although at lower levels than the other three groups immunized with bacteria, respectively. Anti-Actinobacillus actinomycetemcomitans, Bacteroides forsythus and Prevotella intermedia serum antibody levels were also determined and found to be negligible. In conclusion, F. nucleatum immunization does not affect the splenic T cell cytokine response to P. gingivalis. However, F. nucleatum immunization prior to that of P. gingivalis almost completely inhibited the production of anti-P. gingivalis antibodies while P. gingivalis injection before F. nucleatum demonstrated a partial inhibitory effect by P. gingivalis on antibody production to F. nucleatum. The significance of these results with respect to human periodontal disease is difficult to determine. However, they may explain in part differing responses to P. gingivalis in different individuals who may or may not have had prior exposure to F. nucleatum. Finally, the results suggested that P. gingivalis and F. nucleatum do not induce the production of cross-reactive antibodies to other oral microorganisms.

Animals↗

Increase of heat-shock protein and induction of gamma/delta T cells in peritoneal exudate of mice after injection of live Fusobacterium nucleatum.

Fusobacterium nucleatum and Actinobacillus actinomycetemcomitans are Gram-negative rod periodontal pathogens. The peritoneal cavity of Institute of Cancer Research (ICR) mice was used as the local infection model. In vivo production of heat-shock proteins (hsp) was studied by injection of 1/10 minimum lethal dose (MLD) of each live bacteria into mice. Heat-shock proteins 70 and 60 were examined in the extract of peritoneal exudate cells (PEC) from mice injected intraperitoneally with either F. nucleatum or A. actinomycetemcomitans by using sodium dodecylsulphate-polyacrylamide gel electrophoresis and immunoblotting analysis. Although hsp are present in PEC without injection of the bacteria, both hsp increased and reached a peak on day 3 after F. nucleatum injection but not after A. actinomycetemcomitans. Kinetic study of gamma/delta cells in PEC after injection of bacteria showed that the increase of gamma/delta T cells was observed only in the PEC from mice injected with F. nucleatum but not A. actinomycetemcomitans. The gamma/delta T cells in PEC were either CD3+ and CD4+ or CD3+ and CD8+. The differential cell count of PEC suggested that gamma/delta T-cell induction is related to the expansion of the macrophage population. The phagocytic and chemiluminescence responses of macrophages against the same bacteria were compared after intensive immunization with live F. nucleatum and A. actinomycetemcomitans. Elevations of chemiluminescence response and phagocytic function by immunization were observed in the macrophages of mice immunized with F. nucleatum. These results suggest the sequential appearance of hsp, gamma/delta T cells and macrophage activation after fusobacterial infection.

Actinobacillus Infections↗

Minimum inhibitory concentrations for selected antimicrobial agents against Fusobacterium necrophorum isolated from hepatic abscesses in cattle and sheep.

Minimum inhibitory concentrations for 35 antimicrobial agents against 100 Fusobacterium necrophorum isolates from hepatic abscesses in sheep and cattle were determined. Twelve of the thirteen beta-lactam antibiotics tested inhibited growth of 100% of strains tested. Of the remaining antimicrobial agents, extensive susceptibility was found for: spiramycin, josamycin, lincomycin, tylosin, oxytetracycline, chlortetracycline, rufloxacin, metronidazole, cotrimoxazole, sulfadimethoxine, virginiamycin and fosfomycin.

Animals↗

Necrotizing stomatitis associated with Fusobacterium necrophorum in two goats.

Necrotizing stomatitis associated with Fusobacterium necrophorum in two goats is described. Arcanobacterium pyrogenes was the dominant component of the microbial flora cultured under aerobic condition. Treatment with penicillin, at a dose of 50 000 IU/kg for 6 days was used successfully in the cure of the disease.

Animals↗

Fusobacterium nucleatum: a rare cause of bacteremia in neutropenic patients with leukemia and lymphoma.

Although anaerobic bacteremias are uncommon in oncohematologic patients, nevertheless they have been considered an emergent problem in the last few years. Fusobacterium nucleatum is an anaerobic Gram-negative bacillus commonly present in the oral cavity and in the respiratory and genito-urinary tracts. Over a 10-year period 18 episodes of F. nucleatum bacteremia in patients with hematological malignances (15 leukemias and 3 lymphomas) have been observed in our Department of Hematology. Predisposing factors included oropharyngeal mucositis and severe neutropenia owing to intensive chemotherapy. In our experience no septic shock occurred and the outcome of bacteremias caused by F. nucleatum was favorable.

Adolescent↗

Adherence of Fusobacterium necrophorum subsp. necrophorum to ruminal cells derived from bovine rumenitis.

Fusobacterium necrophorum subsp. necrophorum strain VPI 2891 was shown to adhere to the surfaces of ruminal cells derived from bovine rumenitis. The strain also attached to bovine type 1 collagen. Treatment of the bacterium with antiserum to bacterial cells reduced attachment. The bacterial attachment was also markedly reduced when the ruminal cells had been pretreated with anticollagen serum. Fluorescence specific for the collagen was demonstrated on the surface of bovine tissue affected with rumenitis. These findings suggest that F. necrophorum subsp. necrophorum strain VPI 2891 adheres to the ruminal cells derived from rumenitis tissue and that the attachment may be mediated by cellular collagen.

Animals↗

Hemolytic uremic syndrome in an adolescent with Fusobacterium necrophorum bacteremia.

Microorganisms may produce substances that disrupt the interaction between platelets and vascular endothelium, which has been associated with atypical hemolytic uremic syndrome (HUS). We present the first reported case of Fusobacterium necrophorum bacteremia that presented initially with atypical HUS. Antimicrobial therapy eradicated the patient's bacteremia, and plasmapheresis restored platelet-endothelial homeostasis. Understanding the pathophysiologic mechanisms involved in atypical HUS would guide the development of more precise therapies.

Adult↗

Fusobacterium nucleatum in endodontic flare-ups.

OBJECTIVE: The extent to which Fusobacterium nucleatum is recovered from root canals of teeth that present with an interappointment flare-up following endodontic instrumentation was investigated. STUDY DESIGN: Included in the study were 28 patients that sought emergency treatment after initiation of root canal therapy. Only non-painful teeth that had been treated because of a necrotic pulp and periapical inflammatory lesion were studied. Root canal samples for bacterial analysis were taken, transported to a bacteriological laboratory, and processed for a semiquantitative assessment of bacterial isolates. Bacterial findings were correlated with self-assessed pain intensity as recorded by means of a Visual Analogue Scale. Clinical presentation of swelling and presence of exudate in the treated root canals were also linked. RESULTS: Bacteria were recovered from all teeth examined. Gram-negative anaerobic coccoid rods (Prevotella species and Porphyromonas species) were frequent isolates. All teeth in patients who were reported to be in severe pain (Visual Analogue Scale > or = 6) displayed F nucleatum. Nine out of 10 of these teeth also had swelling and exudate in the root canals. Samples from the remaining patients that had teeth with less pain score showed a variable bacterial recovery. None of these teeth displayed F nucleatum. CONCLUSION: F nucleatum appears to be associated with the development of the most severe forms of interappointment endodontic flare-ups.

Acute Disease↗

Fusobacterium necrophorum: a rare cause of meningitis in children.

We report a case of non-fatal meningitis secondary to suppurative otitis media in a previously healthy young boy. Fusobacterium necrophorum was isolated from blood; spinal fluid was cultured aerobically only but yielded no growth. After treatment with intravenous antibiotics and surgery the patient recovered, with a mild sequela of hypoglossus paresis.

Adolescent↗

Primary psoas abscess due to Fusobacterium nucleatum.

A case of primary pyogenic psoas abscess due to Fusobacterium nucleatum is described. Clinicians must maintain a high index of clinical suspicion for the diagnosis of psoas abscess. Although Staphylococcus aureus accounts for most cases of primary psoas abscess, this report emphasizes the importance of bacteriological confirmation of the microorganism involved.

Female↗

Prevalence of Fusobacterium necrophorum and other upper respiratory tract pathogens isolated from throat swabs.

Fusobacterium necrophorum, an anaerobic, Gram-negative rod, has been identified recently as a significant cause of persistent sore throat syndrome (PSTS). This disease is characterised by chronic, recurrent or persistent sore throat, which is believed to respond poorly to penicillin in vivo. The aim of this study is to examine the prevalence of F. necrophorum in all throat swabs received in our diagnostic microbiology department and to compare the results with those for other recognised respiratory pathogens. All throat swabs received in the laboratory over a four-week period were cultured for beta-haemolytic streptococcus groups A, C and G, Corynebacterium diphtheriae, Arcanobacterium haemolyticum and F. necrophorum. Latex agglutination techniques, phenotypic reactions and antibiograms are used to identify these organisms. The age of the patient and the clinical details as stated on the request form were noted. Among a total of 248 samples, 27 were positive for beta-haemolytic streptococcus group A, two were positive for beta-haemolytic streptococcus group C, five were positive for beta-haemolytic streptococcus group G and 24 were positive for F. necrophorum. The most common isolate in the under 20 age group was beta-haemolytic streptococcus group A. In the over 20 age group, F. necrophorum was the pathogen most frequently isolated. A clinical diagnosis of 'sore throat' was most likely to be positive for beta-haemolytic streptococcus group A, a clinical diagnosis of PSTS was most likely to be positive for F. necrophorum and a clinical diagnosis of 'tonsillitis' was equally likely to be caused by beta-haemolytic streptococcus group A or F. necrophorum. beta-haemolytic streptococcus group A was present in 11% of the samples and F. necrophorum was present in 10% of the samples. In total, these two pathogens accounted for 18.5% of throat infections in the sampled group. The results show that F. necrophorum is as significant a cause of throat infection as is beta-haemolytic streptococcus group A. Examination of this provisional data suggests that targeting culture towards these two pathogens may be possible in certain cohorts of patients if more precise clinical data are received from medical staff. However, based on the clinical symptoms routinely provided by clinicians requesting microscopy, culture and sensitivity on throat swabs, F. necrophorum culture is required on all throat swabs received in the laboratory.

Actinomycetaceae↗

Lemierre's disease: postanginal bacteremia and pulmonary involvement caused by Fusobacterium necrophorum.

The combination of acute pharyngotonsillitis, neck pain, fever, and pulmonary septic emboli caused by Fusobacterium necrophorum in a healthy young person is extremely rare. The entity was described by Lemierre in 1936 as a typical syndrome easy to recognize and diagnose exclusively on clinical grounds. A case of Lemierre's disease is reported, and 10 other cases found in the medical literature are reviewed.

Adolescent↗