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PubMed1977

Rabbit polymorphonuclear leukocyte migration in vitro in response to lipopolysaccharides from Bacteroides, Fusobacterium and Veillonella.

Puriified lipopolysaccharides (LPS) from strains of Bacteroides, Fusobacterium and Veillonella incubated with guinea pig serum, were tested for chemotatic activity against rabbit polymorphonuclear leukocytes (PMNs) in modified Boyden chambers. Comparisons were made to a Salmonella LPS (S. enteriditis S-795). Submicrogram amounts of LPS induced positive chemotaxis, and a typical dose-response relationship up to certain dose levels was observed. The difference in chemotactic activity between the Veillonella LPS and LPS-S-795 was not statistically significant. The Fusobacterium LPS showed either a non-significant or a highly significantly lower chemotatic capacity than LPS-S-795. The Bacteroides LPS were also clearly chemotactic, but considerable less when compared to the Salmonella LPS. When serum was not added, the LPS preparation showed no chemotactic activity.

Animals↗

PubMed1977

Rabbit polymorphonuclear leukocyte migration in vivo in response to lipopolysaccharides from Bacteroides, Fusobacterium and Veillonella.

Subcutaneously implanted chambers in rabbits were used for testing the migration of polymorphonuclear leukocytes in response to injected LPS isolated from strains of Bacteroides, Fusobacterium and Veillonella. A salmonella LPS was used as reference endotoxin. No differnece in chemotactic activity between the Veillonella LPS and LPS from Salmoneila was found. Fusobacterium LPS whoed insignificantly lower chemotactic capacity than the Salmonella LPS. The Bacteroides LPS were all significantly less chemotactic than the reference endotoxin. An insignificant correlation between the amount of exudate aspirated from the chambers 5 h after injection of the different LPS preparations and the number of leukocytes per microliter of exudate was found.

Animals↗

PubMed1977

Lethality for mice and chick embryos, pyrogenicity in rabbits and ability to gelate lysate from amoebocytes of Limulus polyphemus by lipopolysaccharides from Bacteroides, Fusobacterium and Veillonella.

Phenol-water extracted lipopolysaccharides (LPS) from Veillonella, Fusobacterium nucleatum, Bacteroides fragilis and Bacteroides melaninogenicus were lethal for mice and 11-days-old chick embryos, pyrogenic in rabbits, and gelated Limulus amoebocyte lysate. Mouse lethality was considerably enhanced by actinomycin-D. In all test systems the endotoxin activity of Veillonella and Fusocbacterium LPS was comparable to that of LPS from Salmonella enteritidis, which was included as a reference endotoxin. The endotoxicity of the Bacteroides LPS was very low. While nanograms of the Veillonella and Fusobacterium LPS killed the chick embryos and gelated the Limulus lysates, microgram amounts of the Bacteroides LPS were needed to give positive reaction in the same test systems. As much as 74 microgram of the most active B. fragilis LPS were required to give a typical biphasic fever response in rabbits. A significant correlation was found between all test results (r = 0.90-0.98, p less than 0.001).

Animals↗

PubMed1983

Evaluation of the Anaerobe-Tek system for the identification of Bacteroides and Fusobacterium species.

Reference strains and clinical isolates of Bacteroides and Fusobacterium species were examined by the Anaerobe-Tek System (A/T-system). Of 104 strain, only 57 (54.8%) were identified correctly to species level. 38 strains (36.5%) were incorrectly identified and for 9 strains (8.7%) there were no codes in the manufacturers' data base manual. The results indicate that, in our hands, the A/T-system in its present form, is not suitable for the identification of clinical isolates of Bacteroides and Fusobacterium species.

Bacteriological Techniques↗

PubMed1977

Hemagglutinating activity of Fusobacterium nucleatum.

Gingival isolates of oral Fusobacterium nucleatum strains (gram-negative anaerobic fusiform bacilli) have shown the characteristic ability to hemagglutinate a variety of erythrocytes (RBC) of human and animal origin. Other members of the genus tested (F. necrophorus, F. varium, and F. mortiferum) displayed little if any ability to hemagglutinate RBC. The hemagglutination (HA) activity could be observed in the F. nucleatum strains with the whole cells and in most instances with sonicated preparations of the organisms. The HA activity was observed in cell wall preparations of the organism and appeared dependent upon a heat-labile protein component of the cell wall. In decreasing order, the RBC that would hemagglutinate with the smallest concentration of HA preparations were rabbit, monkey, human, sheep, horse, and ox. No differences in HA activity of the preparations with cells from the various human blood types were noted. Absorption of the HA preparation of one strain with human cells removed HA moiety was bound to the cells via a Ca2+ binding site interaction since ethylenediaminetetraacetic acid and ethylene glycol-bis-N,N'-tetraacetic acid inhibited binding, and HA could be reestablished by the addition of Ca2+ but not Mg2+. Rabbit antisera to the F. nucleatum strains inhibited HA activity when tested with the HA preparation in the standard test, whereas anti-Leptotrichia buccalis sera or normal rabbit sera had no effect. A tanned-cell passive HA test with rabbit anti-F. nucleatum sera displayed reactivity between the homologous strains but little reactivity with the other Fusobacterium species tested.

Animals↗

PubMed1977

Gliding motility and actinomycin D sensitivity of Fusobacterium nucleatum and other gram-negative rods.

Six strains of gram-negative anaerobic fusiform rods (Fusobacterium and Bacteroides spp.), isolated from deep subgingival locations in humans, were examined for (i) gliding motility in slide cultures, (iii) cell densities on nutrient agar surfaces, and (iii) sensitivity to actinomycin D. Known gliding (FBt) and nonmotile (NM) strains of Myxococcus xanthus served as controls for the gliding and cell density experiments. In the present study, three strains of gram-negative fusiforms from the oral cavity, identified as Fusobacterium nucleatum (1a and P-1) and Bacteroides sp. (P-14), demonstrated characteristics similar to the gliding control strain M. xanthus. The only gliding oral strain sensitive to actinomycin D was a Bacteroides sp. (P-14).

Bacteroides↗

PubMed1989

Inhibition of coaggregation between Fusobacterium nucleatum and Porphyromonas (Bacteroides) gingivalis by lactose and related sugars.

The coaggregation of Fusobacterium nucleatum PK1594 and Porphyromonas (Bacteroides) gingivalis PK1924 was inhibited equally well by lactose, N-acetyl-D-galactosamine, and D-galactose, which caused 50% inhibition of coaggregation at 2 mM sugar concentration. Other sugars such as D-galactosamine, D-fucose (6-deoxy-D-galactose), and alpha-methyl- and beta-methyl-D-galactosides also inhibited coaggregation. Sugar specificity was apparent, since neither L-fucose, L-rhamnose, N-acetyl-D-glucosamine, nor N-acetylneuraminic acid was an inhibitor. Protease treatment of the fusobacterium completely abolished coaggregation, whereas it had no effect on the coaggregating activity of the porphyromonad. Although numerous lactose-inhibitable coaggregating pairs are known to occur among gram-positive bacteria, this report and the accompanying survey (P. E. Kolenbrander, R. N. Andersen, and L. V. H. Moore, Infect. Immun. 57:3194-3203, 1989) are the first studies demonstrating the extensive nature of this type of interaction between gram-negative human oral bacteria. The significance of galactoside-inhibitable coaggregations between these two potential periodontal pathogens is discussed.

Acetylgalactosamine↗

PubMed1989

Isolation of a corncob (coaggregation) receptor polypeptide from Fusobacterium nucleatum.

Corncobs, which are distinct morphological units formed by the ordered coaggregation of a filamentous microorganism and streptococci, can be made in vitro by using oral strains of Fusobacterium nucleatum and Streptococcus sanguis. Previous studies have shown that strains of F. nucleatum contain one of at least two different types of corncob receptor. The objective of this study was to isolate the receptor from F. nucleatum ATCC 10953 as the first step in the elucidation of the molecular basis of corncob formation. The cell envelope fraction from this bacterium was treated with trypsin, delipidated with chloroform-methanol, and subjected to ion-exchange chromatography. A single polypeptide (apparent Mr, 39,500), which was eluted from the column with 0.5 M sodium chloride and extracted with dodecyltrimethylammonium bromide to remove contaminating lipopolysaccharide, inhibited corncob formation between strain ATCC 10953 and S. sanguis CC5A. Similarly derived cell fractions from either F. nucleatum FDC 364 or Fusobacterium necrophorum failed to effect coaggregation in the inhibition assay. Amino acid analysis of the polypeptide showed a moderately hydrophobic character (polarity index, 41) and 11% basic residues. Antiserum made against the purified polypeptide agglutinated F. nucleatum ATCC 10953, neutralized the ability of this bacterium to form corncobs, and agglutinated whole cells of S. sanguis CC5A that were precoated with the receptor polypeptide. The identification and isolation of this receptor should greatly enhance our ability to define some of the complex intergeneric coaggregation mechanisms that are thought to occur in the human oral cavity.

Animals↗

PubMed1991

Sugar metabolism by fusobacteria: regulation of transport, phosphorylation, and polymer formation by Fusobacterium mortiferum ATCC 25557.

Strains of eight Fusobacterium species differed in the ability to use sugars as energy sources for growth. For Fusobacterium russii ATCC 25533, F. gonidiaformans ATCC 25563, and F. nucleatum ATCC 10953 (except for fructose), growth was marginal to poor on all of the sugars tested. Other species displayed reasonable growth on glucose, fructose, mannose, and galactose, and two strains of F. mortiferum (ATCC 25557 and ATCC 9817) grew well on six of the sugars tested, including sucrose and maltose. Glucose transport by resting cells of most of the species was dependent upon (or markedly stimulated by) the presence of a fermentable amino acid. By contrast, F. mortiferum cells rapidly accumulated glucose and other sugars in the absence of amino acids. Although these cells were constitutive for glucose uptake, accumulation of other sugars was specifically induced by growth of F. mortiferum on the appropriate sugar. Spectrophotometric analyses and in situ staining of anionic polyacrylamide gels showed that glucose and fructose (mannose) are phosphorylated by separate ATP-dependent kinases. Fructokinase was stable in air at 4 degrees C, but under these conditions, greater than 70% of the glucokinase activity was lost. After overnight dialysis of the extract, no glucokinase activity was detectable; however, 65% of the initial enzyme activity was retained by inclusion of 1 mM dithiothreitol in the dialysis buffer. Thin-section electron microscopy showed that cells of F. mortiferum produced various amounts of intracellular glycogen during growth on the following sugars (in decreasing order of formation): galactose greater than sucrose greater than glucose greater than mannose greater than fructose. Mechanisms for sugar transport regulation, phosphorylation, and polymer synthesis by F. mortiferum cells are proposed.

Biological Transport↗

PubMed1975

Indirect fluorescent antibody procedure for the rapid detection and identification of Bacteroides and Fusobacterium in clinical specimens.

An indirect fluorescent antibody (IFA) technique was evaluated as a procedure for rapid detection and identification of members of the Bacteroidaceae. Antisera were prepared against 31 members of this family, including species of Bacteroides and Fusobacterium commonly isolated from human infections. The antisera had demonstrated species and/or subspecies specificity. Thirty clinical specimens were studied. Of 13 specimens yielding Bacteroidaceae, for which antisera were available, 23 were presumptively diagnosed by IFA to contain subspecies of B. fragilis and/or Fusobacterium species. Of 17 specimens yielding negative culture results, two were positive by IFA on direct smear. Frequently the in vivo morphology of cells detected in direct smears by this procedure closely mimicked that of cellular debris, tissue cells, and leukocytes. Polyvalent antisera pools facilitated use of the IFA procedure as a practical tool for rapid diagnosis of infections involving the Bacteroidaceae.

Animals↗

PubMed2004

Quantitative microbiological study of subgingival plaque by real-time PCR shows correlation between levels of Tannerella forsythensis and Fusobacterium spp.

A TaqMan-based real-time PCR assay was established to quantify the periodontopathic bacteria Tannerella forsythensis and Fusobacterium spp. With this assay, the prevalence and proportion of these bacteria in clinical specimens were evaluated. Our preliminary results suggest a positive colocalization of T. forsythensis and Fusobacterium spp. in periodontal pockets.

Adult↗

PubMed2024

Development of SacB-based counterselection for efficient allelic exchange in Fusobacterium nucleatum.

Fusobacterium nucleatum, prevalent in the oral cavity, is significantly linked to overall human health. Our molecular comprehension of its role in oral biofilm formation and its interactions with the host under various pathological circumstances has seen considerable advancements in recent years, primarily due to the development of various genetic tools for DNA manipulation in this bacterium. Of these, counterselection-based unmarked in-frame mutation methods have proved notably effective. Under suitable growth conditions, cells carrying a counterselectable gene die, enabling efficient selection of rare, defined allelic exchange mutants. The sacB gene from Bacillus subtilis, encoding levansucrase, is a widely used counterselective marker partly due to the easy availability of sucrose. Yet, its potential application in F. nucleatum genetic study remains untested. We demonstrated that F. nucleatum cells expressing sacB in either a shuttle or suicide plasmid exhibit a lethal sensitivity to supplemental sucrose. Utilizing sucrose counterselection, we created an in-frame deletion of the F. nucleatum tonB gene, a critical gene for energy-dependent transport processes in Gram-negative bacteria, and a precise knock-in of the luciferase gene immediately following the stop codon of the hslO gene, the last gene of a five-gene operon possibly related to the natural competence of F. nucleatum. Post-counterselection with 5% sucrose, chromosomal plasmid loss occurred in all colonies, leading to gene alternations in half of the screened isolates. This sacB-based counterselection technique provides a reliable method for isolating unmarked gene mutations in wild-type F. nucleatum, enriching the toolkit for fusobacterial research.IMPORTANCEInvestigations into Fusobacterium nucleatum's role in related diseases significantly benefit from the strategies of creating unmarked gene mutations, which hinge on using a counterselective marker. Previously, the galk-based allelic exchange method, although effective, faced an inherent limitation-the need for a modified host. This study aims to surmount this limitation by substituting galK with sacB for gene modification in F. nucleatum. Our application of the sacB-based methodology successfully yielded a tonB in-frame deletion mutant and a luciferase gene knock-in at the precise chromosomal location in the wild-type background. The new method augments the existing toolkit for F. nucleatum research and has far-reaching implications due to the easy accessibility to the counterselection compound sucrose. We anticipate its broader adoption in further exploration, thereby reinforcing its critical role in propelling our understanding of F. nucleatum.

Fusobacterium nucleatum↗

PubMed1992

Immunoglobulins in milk from cows immunized with oral strains of Actinomyces, Prevotella, Porphyromonas, and Fusobacterium.

Immunization of pregnant cows with bacteria leads to the presence of high concentrations of specific antibodies in colostrum and milk. A total of 14 cows was immunized with single strains of heat-killed oral bacteria or pools of strains of Actinomyces, Porphyromonas, Prevotella, and Fusobacterium. Two cows were treated with adjuvant alone. The mean percentages of IgG1, IgG2, IgM, and IgA in all of the milks were 83.8, 3.8, 9.3, and 3.1, respectively. ELISA and whole cell agglutination assays demonstrated high titers in the milks from the cows immunized with either individual strains or the bacterial pools. The highest titers determined by ELISA belonged to the IgG1 isotype and in several milks were 64-fold greater than titers in milk from cows treated with adjuvant alone. The concentrations of all antibodies and the titers determined by ELISA and whole cell agglutination assays markedly decreased from the first to the sixth milkings. The functional specificity of the antibodies was demonstrated by agglutination tests against a wide range of bacteria including members of Actinomyces, Fusobacterium, Porphyromonas, Prevotella, Streptococcus, Eubacterium, Propionibacterium, Peptostreptococcus, Bacteroides, Actinobacillus, Haemophilus, Capnocytophaga, and Wolinella. Minimal cross-reactions with bacteria in other genera were observed with all of the milks. High-titer milk preparations have been obtained from immunized cows, and the capacity of the bovine antibodies to agglutinate target bacteria indicates their potential usefulness in oral passive immunization studies.

Actinomyces↗

PubMed1986

[Post-angina septicemia caused by Fusobacterium necrophorum in a 7-year-old child].

A fusobacterium necrophorum septicemia due to a neglected peritonsillar abscess is reported in a 7 year-old boy with no significant past medical history. Osteo-articulary, hepatic and pleuro-pulmonary septic localizations, with an otherwise favourable outcome left severe orthopedic sequelae in the right hip. This resembles the post-peritonsillar abscess septicemia described by Lemierre in 1936 which was due to an anaerobic bacillus (fusobacterium). The reappearance of this pathology should lead to systematic anaerobic blood and abscess studies. Penicillin G and Metronidazole are still efficient in controlling this organism.

Child↗

PubMed1986

Anaerobic septicemia secondary to Fusobacterium mortiferum.

A case of anaerobic sepsis associated with Fusobacterium mortiferum is reported. Blood cultures from a 60-year-old man with type II diabetes mellitus, hypertension, severe atherosclerotic cardiovascular disease, and renal insufficiency revealed on a gramstained smear highly pleomorphic gram-negative bacilli with bizarre forms and round bodies. Growth of the organism on nonselective anaerobic media and analysis of its pattern produced results characteristic of Fusobacterium mortiferum.

Fusobacterium Infections↗

PubMed1983

Comparative biological features of a rat liver abscess model induced with three Fusobacterium necrophorum strains.

Several biological features were compared in a rat liver abscess model, using intraportal inoculations with 3 bovine strains of Fusobacterium necrophorum which varied in virulence. Serum alanine aminotransferase activities were increased significantly (P less than 0.05) in rats inoculated with F necrophorum 2101 by postinoculation hours 6, 12, and 24. Thereafter, alanine aminotransferase values returned to base line for the remainder of the experiment. Also, rats inoculated with F necrophorum 2101 had a significantly greater (P less than 0.05) weight loss than did the control rats during the first 5 postinoculation days and developed leukocytosis characterized by a neutrophilia with a left shift. The duration of the bacteremia was related directly to the virulence of the F necrophorum strain. Fusobacterium necrophorum 2101, a biotype A which was the most virulent, induced the most persistent bacteremia; F necrophorum 2035, a biotype B which was the least virulent, produced the shortest bacteremia; and F necrophorum 2030, a biotype AB which was of intermediate virulence, led to bacteremia of intermediate duration. Plasma endotoxin was demonstrated intermittently during the first 24 hours, but did not correlate with the bacteremia.

Alanine Transaminase↗

PubMed1982

[Comparative study of different substrates as the nutrient base of a culture medium for Bacteroides and Fusobacterium].

The results of the comparative study of different substrates prepared from foodstuffs and inedible raw materials and intended for use as the nutrient base of a medium for cultivating the organisms of the genera Bacteroides and Fusobacterium have demonstrated that commonly used nutrient bases prepared from meat, fish, casein and inedible animal blood (blood substitutes) are suitable for this purpose. The dried nutrient base prepared from the mixture of outdated blood substitutes (hydrolysin and casein hydrolysate) (see text for Russian letters) has been found to ensure the optimum growth of Bacteroides and Fusobacterium.

Animals↗

PubMed1998

[Investigations into the occurrence and the antibiotic susceptibility of gram negative anaerobes of the genera Bacteroides, Prevotella, Porphyromonas and Fusobacterium in specimens obtained from diseased animals].

From different samples of 247 diseased animals (cattle, sheep, goat, horse, pig, dog, cat, rodent, zoo-animals), 410 strains of gram-negative anaerobes were cultured. 297 isolates (72.4%) could be differentiated to the species level by using cultural-biochemical methods, gaschromatography and cell-wall-lipidanalysis. They belonged to 29 different species. For an additional 113 strains (27.6%) only the genus could be determined. Bacteria belonging to the genus Fusobacterium occurred with the highest isolation rates (36% of all strains) in the samples examined, followed by Bacteroides spp. (26.1%), Prevotella spp. (19.9%) and Prophyromonas spp. (17.8%). Fusobacterium necrophorum was the single species isolated most frequently. Antibiotic susceptibility tests by E-test were performed on 100 strains belonging to the the above mentioned genera. Of these strains 18% were resistant to penicillin and 20% to tetracycline. The resistant strains belonged mainly to the Bacteroides fragilis-group. Resistant rates to most other antimicrobial agents tested were Amoxicillin in combination with clavulanic acid: 1%, Chloramphenicol: 3%, Clindamycin:8%. All 100 selected strains proved to be susceptible to Metronidazol.

Animals↗