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PubMed1991

Partial characterization of Fusobacterium necrophorum protease.

Partial characterization of Fusobacterium necrophorum protease was investigated. The protease was partially purified by gel filtration with Toyopearl HW 55. The final preparation was inactivated completely by heating at 60 degrees C for 30 min and inhibited by ascorbic acid, sodium thioglycollate and p-hydromercuribenzoate. Antibody response to the protease was demonstrated in mice receiving 10(4) CFU of F. necrophorum.

Antibodies, Bacterial↗

PubMed1987

Multiple Fusobacterium nucleatum liver abscesses. Association with a persistent abnormality in humoral immune function.

A previously healthy 29-year-old man developed multiple hepatic abscesses secondary to Fusobacterium nucleatum. No underlying local disease was found. The leading portal of entry for the bacterium may have been the oral cavity; 4 days before the onset of his illness he had had extensive dental work. Immunological evaluation during the illness and in late convalescence (16 weeks after onset) revealed a persistent B cell abnormality characterized by markedly depressed in vitro secretion of immunoglobulins in response to pokeweed mitogen; however, serum immunoglobulins and IgG subclasses were normal. Abnormal numbers of suppressor T cells (OKT8+) and increased suppressor function were also present. Anaerobic pyogenic liver abscesses may occur in the absence of obvious underlying disease, but this case suggests that there may be an association with in vitro abnormalities of the immune system.

Adult↗

PubMed1977

Inhibition of leukocyte migration by the agarose plate technique. Application to antigen from Candida albicans and Fusobacterium nucleatum.

Leukocyte migration inhibition by antigen from Candida albicans and Fusobacterium nucleatum was studied in man by means of the direct leukocyte migration agarose technique (LMAT). Antigens were prepared by prolonged ultrasonication and were added to the leukocyte cultures in the original as well as in the concentrated solutions. Significant inhibition of migration with a 10-fold concentrated solution of candida antigen was demonstrated in subjects showing apositive intracutaneous reaction to candida. With the fusobacterial antigen, inhibition of migration or stimulation of migration were demonstrated in subjects with slight peridontal inflammation. A 5-fold increase in the concentration of fusobacterial antigen resulted in increased stimulation of migration or inhibition of migration. Preincubation of the leukocytes with puromycin abolished both inhibition and stimulation of migration by candidal or fusobacterial antigen. The study confirms that it is important to study dose-related effects when applying the LMAT to test cell-mediated immunity. The study is in support of stimulation of migration being an immunologic reaction.

Antigens, Fungal↗

PubMed1978

Biological and biochemical characteristics of Fusobacterium necrophorum leukocidin.

The production of a leukocidal exotoxin by 18-hour dialysis cultures of Fusobacterium necrophorum was detected in vitro by a cytotoxicity assay, using bovine leukocytes isolated from peripheral blood. Biological characteristics were determined while maximizing the in vitro assay. Biochemical characteristics of heat stability and stability to degradative enzymes were investigated and indicated that the leukocidin is protein in nature. The characteristics indicate that this leukocidin is different from previous descriptions in the literature of the characteristics of a leukotoxin produced by F necrophorum.

Biological Assay↗

PubMed1988

Synergistic effects of Fusobacterium necrophorum lipopolysaccharide, cytoplasmic, and culture supernatant fractions on induction of acute hepatic necrosis in rabbits.

Synergistic effects of toxic fractions of Fusobacterium necrophorum were examined for evaluation of the role of the toxin in inducing liver abscesses in rabbits. Cytoplasmic and culture supernatant fractions of F necrophorum had preparative activity for the Shwartzman reaction, and lipopolysaccharide of F necrophorum had preparative and provocative activities for the reaction. All 3 fractions were hepatotoxic. Inoculation into the bile duct with each fraction followed by intravenous inoculation with F necrophorum or Escherichia coli lipopolysaccharide had a greater synergistic effect in inducing severe hepatic necrosis than did inoculations with double doses of the cytoplasmic or supernatant fractions of F necrophorum. This synergism may have been attributable to the Shwartzman reaction.

Acute Disease↗

PubMed1986

[Evaluation of the sensitivity to antibiotics of Bacteroides and Fusobacterium using the ATB ANA system].

The susceptibility to antibiotics of 100 Gram negative obligate anaerobes of the Bacteroides and Fusobacterium species was tested using the ATB ANA method. Results were compared to those obtained by addition of the tested antibiotics to Wilkins-Chalgren medium in the critical concentrations. Agreement between the two methods was satisfactory (80 to 100%) for the low cutoff concentrations, except for doxycycline. When the pathogens are divided into susceptible, intermediate and resistant strains, results are more difficult to interpret because of the existence of the intermediate category: however, major discrepancies (R/S) did not exceed 3.2% except for cefotaxime (15%). Our results show that use of ATB ANA microplates in an anaerobic enclosure is a valuable method in clinical practice.

Anti-Bacterial Agents↗

PubMed1986

Serological response of mice to Fusobacterium necrophorum.

Mice immunised with killed or living Fusobacterium necrophorum, by five different regimens, almost invariably failed to produce antibodies demonstrable by a passive haemagglutination test. An enzyme-linked immunosorbent assay (ELISA), however, usually demonstrated a serum antibody response. This suggested that F necrophorum was not in fact immunosuppressive in mice--a possibility that had been entertained to explain the great difficulty in protecting mice immunologically against challenge with F necrophorum.

Animals↗

PubMed1985

Identification of common antigens in ribosome-rich extracts from Fusobacterium necrophorum.

Ribosome-rich extracts (RRE) were prepared by differential and ultracentrifugation from 25 bovine and 6 ovine isolates of Fusobacterium necrophorum (FN) including both biotypes A and B. A pooled rabbit antiserum was prepared against whole-cell and sonicated whole-cell bacterins of F necrophorum isolate FN 3080, and a 2nd pooled rabbit antiserum was prepared against a RRE of FN 3080. The RRE of the 25 bovine isolates were tested against the FN 3080 whole-cell antiserum, using Ouchterlony double-immunodiffusion procedures. One to 3 precipitin lines were observed with the 25 isolates. The individual bovine isolates were found to have lines of identity with 5 to 21 of the other 24 isolates. The 25 bovine isolates and the 6 ovine isolates were then compared, using the FN 3080 RRE antiserum. One to 3 precipitin lines were observed for the 31 isolates with the RRE antiserum, and lines of identity were observed between all 31 of the isolates. These results indicated that common antigens are present in the RRE from a wide variety of F necrophorum isolates including both A and B biotypes.

Animals↗

PubMed1985

Development of enzyme-linked immunosorbent assays for the detection of Fusobacterium necrophorum antibody in animal sera.

Enzyme-linked immunosorbent assays (ELISA) for the detection of Fusobacterium necrophorum antibody in the sera of rabbits, cattle, and sheep were developed, using a ribosome-rich extract (RRE) from F necrophorum as the antigen. Test character, including optimal antigen dilution and substrate incubation periods, was established, using rabbit, bovine, and ovine antisera produced against RRE from isolates of F necrophorum. Rabbit antisera produced against 7 other species of bacteria were used to test the specificity of the F necrophorum RRE antigen. Cross-reactivity was not detected. Sera from 50 feedlot cattle were examined with the bovine ELISA. Of the 50 samples, 43 (88%) were positive for F necrophorum antibody. The ELISA developed in this study were sensitive and specific and appear to be readily adaptable to serologic investigations of F necrophorum.

Animals↗

PubMed1979

Hemolytic activity of Fusobacterium necrophorum culture supernatants due to presence of phospholipase A and lysophospholipase.

Culture supernatants of Fusobacterium necrophorum demonstrated hemolytic activity. The hemolysin(s), which was partially purified by ammonium sulfate precipitation, was temperature-dependent and heat labile. The spectrum of hemolytic activity against various erythrocytes included rabbit, human, and dog erythrocytes. Goats, sheep, and bovine erythrocytes showed only trace hemolysis. According to results of thin-layer chromatography, the hemolysin hydrolyzed rabbit erythrocyte phosphatidyl choline, phosphatidyl ethanolamine, lysophosphatidyl choline, and bovine phosphatidyl choline. Hydrolysis of egg yolk phosphatidyl choline, bovine phosphatidyl ethanolamine, cholesterol, 1,2-dipalmitin, 1,3-dipalmitin, sphingomyelin, or triolein was not detected by thin layer chromatography. A more sensitive procedure utilizing gas-liquid chromatography revealed that, of the substrates tested, the following were bein hydrolyzed: bovine and egg yolk phosphatidyl choline, lysophosphatidyl choline, alpha-palmito-beta-eleoyl-L-alpha lecithin and alpha-oleoyl-betal-palmitoyl-L-alpha lecithin. Substrates which were weakly hydrolyzed were bovine phosphatidyl ethanolamine, DL-alpha-hosphatidyl ethanolamine dipalmitoyl, 1,2-dipalmitin, 1,3-dipalmitin, and triolein.

Chromatography, Gas↗

PubMed1979

Correlations between leukocidin production and virulence of two isolates of Fusobacterium necrophorum.

Leukocidin production by Fusobacterium necrophorum was suggested to be an important element in the development of intraabdominal and liver abscesses in mice. Leukocidin production by cultures of F necrophorum was demonstrated by an in vitro assay. One of two isolates of F necrophorum was demonstrated to produce leukocidin. The leukocidin-producing strain was observed to be more infective than the nonleukocidin-producing strain (as demonstrated by abscess formation following intraperitoneal injection of immune-suppressed and normal mice). The infectivity of the leukocidin-producing strain was increased by successive passage in immune-suppressed mice. A simultaneous increase in leukocidin production was also demonstrated. The nonleukocidin-producing strain could not be passed effectively and was relatively noninfective for mice.

Animals↗

PubMed1977

Susceptibility to erythromycin of anaerobes of the genera Bacteroides, Fusobacterium, Sphaerophorus, Veillonella, Clostridium, Corynebacterium, Peptococcus, Peptostreptococcus.

The minimal inhibitory concentrations (MIC) of erythromycin were determined by broth dilution tests for 313 anaerobic strains, most of which were clinical isolates. All the gram-positive anaerobes tested (84 Peptococcaceae, including 21 Peptostreptococcus anaerobius and 15 Peptococcus variabilis; 65 Corynebacterium acnes and 29 Clostridium strains, including 13 C. perfringens) were sensitive (MIC values 0.012 through 3.12 microgram erythromycin/ml); so were 111 cultures of gram-negative anaerobes (52 Bacteroides fragilis, 12 B. thetaiotaomicron, 7 B. vulgatus, 13 B. oralis, 4 B. melaninogenicus, 10 Sphaerophorus necrophorus, 2 Veillonella sp., 11 members of other species). Erythromycin at concentrations of 6.25 through 200.0 microgram/ml was active against 24 strains (1 B. fragilis, 4 Fusobacterium fusiforme, 9 Sph. freundi, 10 Sph. varius). The present results are compared to the limited number of reports existing with regard to the susceptibility of anaerobes to erythromycin.

Bacteroides↗

PubMed1978

Swine dysentery: studies of gnotobiotic pigs inoculated with Treponema hyodysenteriae, Bacteroides vulgatus, and Fusobacterium necrophorum.

Transmission experiments were carried out in gnotobiotic pigs to determine whether lesions typical of swine dysentery could be produced by oral inoculation of Treponema hyodysenteriae in combination with Bacteroides vulgatus or Fusobacterium necrophorum, or both. Each of the organisms had been isolated from swine with early lesions of the disease. Lesions were not found in 6 pigs inoculated with T hyodysenteriae alone, in 4 pigs given F necrophorum and T hyodysenteriae, or in 4 pigs given B vulgatus and F necrophorum. Lesions typical of swine dysentery developed in 8 pigs given B vulgatus, F necrophorum, and T hyodysenteriae as well as in 3 of 4 pigs given B vulgatus and T hyodysenteriae. In both of these groups, the inoculated bacteria were recovered from the colon, and T hyodysenteriae was demonstrated in the colonic crypts, epithelium, and lamina propria. The pathogenicity of the T hyodysenteriae was shown by the development of characteristic signs and lesions of swine dysentery in 12 of 14 naturally farrowed pigs inoculated with T hyodysenteriae alone.

Animals↗

PubMed1980

Induction of Epstein-Barr virus antigens in human lymphoblastoid P3HR-1 cells with culture fluid of Fusobacterium nucleatum.

Epstein-Barr virus-associated early antigen and viral capsid antigen were efficiently induced in human lymphoblastoid P3HR-1 cells with culture fluid of Fusobacterium nucleatum, a member of the indigenous microbial flora of the human host. This finding may suggest a new approach to assess the possible role of the "cofactor(s)" in the etiology of Epstein-Barr virus-related diseases.

Antigens, Viral↗

PubMed1983

Antitumor activity in the culture supernatant fluid of Fusobacterium nucleatum strain KO-31.

Antitumor activity of Fusobacterium nucleatum strain KO-31, which had been isolated from the oral cavity, was investigated in Ehrlich carcinoma-bearing mice. The strain KO-31 was cultured in TF medium for 48 hr. One-half ml of the culture supernatant fluid was injected intraperitoneally during a 7-day period to mice with Ehrlich ascites carcinoma. The survival time of the treated mice prolonged remarkably. A significant reduction in size of the solid Ehrlich tumor was also achieved with the supernatant. Antitumor activity was not observed in the cell extract.

Animals↗

PubMed1984

Antitumor activity of sediment fractions from Fusobacterium nucleatum culture supernatant.

The present study was carried out to examine antitumor activity of extracts from Fusobacterium nucleatum culture supernatant. The extracts were prepared as sediment fractions by ethanol precipitation, isoelectric point precipitation and proteolytic enzyme-treatment. An ethanol fraction obtained by addition of 60% ethanol to the culture supernatant showed effective antitumor activity to ascites and solid type tumor of Ehrlich carcinoma and also to solid type tumor of Sarcoma 180 in ICR mice. A sediment fraction extracted by adjusting an ethanol fraction to pH 4.0 was also found to give distinctive antitumor activity to Ehrlich ascites carcinoma. This fraction was further treated with pronase to eliminate free nucleotides, and then ultrafiltrated. The pronase-treated fraction gave better antitumor activity to both ascites and solid type tumor of Ehrlich carcinoma or Sarcoma 180. The ratios (%) indicating antitumor activity ranged from 131 to 188% even at 4-8 weeks after the tumor cell inoculation.

Animals↗

PubMed1984

Purification and characterization of C-S lyase from Fusobacterium varium. A C-S cleavage enzyme of cysteine conjugates and some S-containing amino acids.

An enzyme responsible for the carbon-sulfur bond cleavage of various S-aryl, S-aralkyl, and S-alkyl cysteines has been purified about 270-fold from Fusobacterium varium. Incubation of a cysteine conjugate of p-bromobenzene with the enzyme yielded equimolar amounts of p-bromobenzenethiol, pyruvic acid, and ammonia, indicating that the carbon-sulfur bond cleavage proceeds via an alpha, beta-elimination reaction. The enzyme activity was inhibited either by hydroxylamine or KCN and stabilized by pyridoxal phosphate, which probably acted as cofactor. The broad substrate spectrum of this enzyme suggested an important role of the intestinal microflora in the in vivo formation of methylthio-containing metabolites of various xenobiotics.

Chromatography, DEAE-Cellulose↗

PubMed1982

Induction of immunologic memory by a lipopolysaccharide-protein complex isolated from Fusobacterium necrophorum: cellular response.

The ability of a lipopolysaccharide protein (LPS-P) complex extracted from Fusobacterium necrophorum to establish immunologic memory in BALB/c mice splenocytes was demonstrated. The LPS-P molecule differed from the phenol water-extracted LPS because it contained approximately 12% protein. Initial experiments showed that primary and secondary spleen plaque-forming cell (PFC) responses to IV or IM injections of LPS-P were highly dose-dependent. Suitable primary doses stimulated significant (P less than 0.05) amounts of direct and direct + indirect PFC by postinoculation day (PID) 14 and primed the mice for an enhanced secondary response to small booster injections. When mice were inoculated with a suitable primary IM dose of aluminum hydroxide-precipitated LPS-P, significant amounts of direct and direct + indirect PFC were detectable through PID 120. Moreover, significant enhancement of these values was attained with an IV booster injection at PID 105. Primary IV inoculation with LPS-P produced similar results, although the primary response was not as persistent.

Animals↗