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PubMed1988

Annual incidence, epidemiology, and comparative in vitro susceptibilities to cefoxitin, cefotetan, cefmetazole, and ceftizoxime of recent community-acquired isolates of the Bacteroides fragilis group.

The six species of the Bacteroides fragilis group are potent pathogens and commonly have different susceptibility patterns. We determined the relative annual isolation rate of anaerobic bacteria and the susceptibility of B. fragilis group species isolated during 1987 at two community hospitals. The relative frequencies of isolation of 261 strains were as follows: B. fragilis, 61%; B. thetaiotaomicron, 17%; B. distasonis, 7%; B. vulgatus, 6%; B. ovatus, 5%; and B. uniformis, 4%. A total of 234 recent clinical isolates were tested against cefmetazole, cefotetan, cefoxitin, ceftizoxime, clindamycin, imipenem, and piperacillin by a brucella agar dilution method. Imipenem was the most active agent tested with all but three isolates (two B. vulgatus and one B. distasonis) susceptible to less than 2 micrograms/ml. Of the cephalosporins tested, cefoxitin, cefotetan, and cefmetazole were relatively equal against B. fragilis, with 93 to 98% of strains susceptible to 32 micrograms/ml. Ceftizoxime was less active, with an MIC for 90% of strains tested of 128 micrograms/ml and only 75% of isolates susceptible to 32 micrograms/ml. Against B. ovatus, B. vulgatus, B. thetaiotaomicron, and B. uniformis, cefoxitin showed a two- to threefold-superior activity compared with that of cefotetan and cefmetazole. In general, ceftizoxime was much less active, except against B. distasonis, for which 78% of isolates were susceptible to 32 micrograms/ml compared with 68% for cefoxitin, 19% for cefmetazole, and 16% for cefotetan. Clindamycin and piperacillin showed activity similar to that of cefoxitin, except piperacillin was less active versus B. vulgatus and B. distasonis. We therefore suggest that clinical laboratories determine the species of B. fragilis group isolates as well as perform susceptibility studies on the isolates. Clinicians should be aware that while B. fragilis is the most frequent isolate, 38% of isolates are from other, more resistant B. fragilis group species.

Bacteroides Infections↗
PubMed1989

Diarrheal disease caused by enterotoxigenic Bacteroides fragilis in infant rabbits.

Enterotoxigenic Bacteroides fragilis caused severe, nonhemorrhagic, watery diarrhea when 10(9) CFU of a porcine or human isolate was administered orogastrically to 3-day-old rabbits. The bacterium colonized the intestinal tract with a predilection for the large intestine (10(9) CFU/g of cecal contents). Diarrhea occurred at an average of 4.6 days postinoculation, and 84% of rabbits were dead or moribund at an average of 8.8 days postinoculation. The disease was characterized by watery diarrhea and dehydration. Severe histologic lesions including inflammation, exfoliation of epithelial cells, and crypt hyperplasia were observed throughout the colon. There was no indication of bacteremia or of bacterial adherence to or invasion of intestinal epithelial cells. Rabbits inoculated with nonenterotoxigenic B. fragilis were colonized with B. fragilis but did not develop clinical disease or intestinal lesions. While the pathogenesis of this disease is undefined, clinical signs of disease and histologic changes were consistent with a mechanism of net secretion of fluid into the small intestine and decreased absorption of fluid from the large intestine. Enteric disease caused by enterotoxigenic B. fragilis in infant rabbits was similar to naturally occurring enteric disease associated with the bacterium in humans and livestock. This study established that enterotoxigenic B. fragilis is enteropathogenic in intact infant rabbits.

Animals↗
PubMed1990

Rabbit model to evaluate enterovirulence of Bacteroides fragilis.

Enterotoxigenic Bacteroides fragilis elicited fluid accumulation in the intestinal tract and exfoliation of epithelial cells in the proximal colon of the 2-week-old rabbit. The rabbit model was employed to screen isolates of B. fragilis for enterovirulence and to study the pathogenesis of disease caused by enterotoxigenic B. fragilis.

Animals↗
PubMed1976

Serogrouping of Bacteroides fragilis subsp. fragilis by the agglutination test.

The agglutination technique was used to establish a serological classification scheme for 98 strains of Bacteroides fragilis subsp. fragilis isolated from clinical specimens and normal human feces. Absorbed antisera were prepared to seven strains of B. fragilis subsp. fragilis. These seven absorbed antisera were species as well as subspecies specific and provided the basis of the serological classification scheme. This scheme was composed of 21 serogroups; seven of these serogroups contained only one group component. There was a total of 45 serological patterns. This serological scheme may be used for the serological classification of strains of B. fragilis subsp. fragilis and to study the epidemiology of this organism.

Agglutination Tests↗
PubMed1992

Analysis of rRNA restriction fragment length polymorphisms from Bacteroides spp. and Bacteroides fragilis isolates associated with diarrhea in humans and animals.

The Escherichia coli rRNA operon rrnB was used as a 32P-labeled hybridization probe in Southern blots of genomic DNAs from representative strains of the saccharolytic, gram-negative, obligate anaerobes of the genus Bacteroides. Control experiments with the B. fragilis type strain ATCC 25285 established that nearly identical rRNA fragment patterns were produced when either the E. coli rrnB gene probe or homologous rRNA isolated from B. fragilis was used as the probe. In addition, it was shown that a specific 16S or 23S rrnB gene probe also could be used to produce fragment patterns suitable for analysis. Thirty-one strains from 8 of the 10 recognized Bacteroides species were then examined. The resulting autoradiographs revealed specific fragment patterns for all but one (B. ovatus) of the species tested. Restriction fragment length polymorphisms were observed for many of the strains tested, but these differences did not hinder species classification. The five B. ovatus strains examined did not form a distinct group, and their rRNA fragment patterns displayed a marked heterogeneity. The same approach was applied to a unique set of enterotoxin-producing B. fragilis strains isolated from animals and humans with diarrhea. The results demonstrated that these strains were in fact B. fragilis and that they produce rRNA fragment patterns closely related to those of the type strain ATCC 25285. This set of strains did not appear to form a separate subgroup or genotype within the B. fragilis species, and there were no distinguishable restriction fragment length polymorphisms that could be used to specifically separate enterotoxin-producing strains from nonenterotoxigenic strains.

Animals↗
PubMed1997

Detection of enterotoxigenic Bacteroides fragilis by PCR.

Strains of enterotoxigenic Bacteroides fragilis (ETBF) are associated with diarrhea in young farm animals and, at least in particular settings, in children. Enterotoxin production by ETBF is currently detected by a tissue culture assay with HT-29 cells. We have developed a PCR assay based on the detection of the enterotoxin gene to identify ETBF in culture and in stool samples. Overall, 113 bacterial strains were examined, including 3 B. fragilis reference strains, 75 B. fragilis isolates (comprising 40 ETBF isolates), 20 Bacteroides spp. other than B. fragilis, and 15 strains belonging to other genera. Complete agreement was found between the results of the tissue culture assay and those of the PCR for our strains. PCR was also used to detect ETBF directly in fecal samples. Stools from two healthy volunteers were spiked with known numbers of ETBF and were processed by three different methods. A culture method, which required inoculation of the stools on selective plates and the collection of the whole bacterial growth ("sweeps"), was found to be the most sensitive. PCR performed with the plate sweeps yielded amplification products with a detection limit of 10(5) to 10(4) CFU/g of feces. By this method 18 samples of diarrheic stools (10 positive and 8 negative for ETBF) were examined. The results of the PCR were in accordance with the culture results in all cases. The proposed PCR assay represents a diagnostic tool for the rapid identification of ETBF in culture as well as in fecal samples.

Bacterial Toxins↗
PubMed1999

Prevalence of enterotoxigenic Bacteroides fragilis in children with diarrhea in Japan.

In age-matched controlled studies performed in Japan, enterotoxigenic Bacteroides fragilis was isolated from 14.9% of 114 children aged 1 to 14 years with antibiotic-unassociated diarrhea (AUD) and 6.5% of 108 children aged 1 to 6 years with antibiotic-associated diarrhea (AAD). The difference in comparison with control children, was significant for AUD children but not AAD children.

Adenoviruses, Human↗
PubMed2000

The alleles of the bft gene are distributed differently among enterotoxigenic Bacteroides fragilis strains from human sources and can be present in double copies.

Enterotoxigenic Bacteroides fragilis (ETBF) strains are associated with diarrheal disease in children. These strains produce a zinc metalloprotease enterotoxin, or fragilysin, that can be detected by a cytotoxicity assay with HT-29 cells. Recently, three different isoforms or variants of the enterotoxin gene, designated bft-1, bft-2, and bft-3, have been identified and sequenced. We used restriction fragment length polymorphism analysis of the PCR-amplified enterotoxin gene to detect the isoforms bft-1 and bft-2 or bft-3 borne by ETBF. By sequencing the portion of the bft gene corresponding to the mature toxin in some strains and applying allele-specific PCR for strains categorized as bft-2 or bft-3, we found in our collection two strains harboring bft-3, a variant that had been described for isolates from East Asia. Analysis of 66 ETBF strains from different sources showed that bft-1 is the most frequent allele, being present in 65% of isolates; it is largely predominant in isolates from feces of adults, while bft-2 is present in isolates from feces of children. This association is statistically significant (P, 0.0064). Sixteen strains were examined by Southern hybridization using, as probes, the bft and second metalloprotease genes, both included in a pathogenicity islet. Five strains were found to harbor double copies of both genes, suggesting that the whole islet was duplicated. Four of these strains, harboring bft-1 (three strains) or bft-2 (one strain), were found to produce a large amount of biologically active toxin, as determined by a cytotoxicity assay with HT-29 cells. The strains harboring bft-3, either in a single copy or in double copies, produced the smallest amount of toxin in our collection.

Adult↗
PubMed2002

In vitro susceptibilities of the Bacteroides fragilis group species: change in isolation rates significantly affects overall susceptibility data.

A comparison of antimicrobial susceptibility data of species of the Bacteroides fragilis group for 1989-1990 and 1998-1999 studies showed statistically significant increases or decreases in in vitro activity. Overall significant increases in resistance were noted for ampicillin-sulbactam and clindamycin, while significant decreases in resistance were noted for ertapenem and cefoxitin. Susceptibilities to piperacillin-tazobactam, imipenem, meropenem, and trovafloxacin remained virtually the same for the two studies. Importantly, a change in the rates of isolation of the various species showed the B. fragilis species comprised 58% of the isolates in 1989 to 1990 and 45% of the isolates in 1998 to 1999. This change in rates of isolation of B. fragilis versus non-B. fragilis species had an overall effect on susceptibility data.

Anti-Bacterial Agents↗
PubMed1978

New medium for selection and presumptive identification of the Bacteroides fragilis group.

A medium, Bacteroides fragilis bile-esculin (BBE) agar, was designed for the selection and, presumptive identification of the B. fragilis group. BBE agar contains bile, esculin, ferric ammonium citrate, hemin, and gentamicin in a Trypticase soy agar base. Growth in the presence of 20% bile and esculin hydrolysis, detected by blackening of the medium, provide presumptive evidence for the identification of the B. fragilis group. In addition to stimulating the growth of many strains of the B. fragilis group, hemin provides the option of testing isolates for catalase production. Gentamicin and bile prevent the growth of most organisms other than the esculin-positive bacteroides that can tolerate bile. Of 160 clinical isolates of the B. fragilis group tested on BBE agar, 159 grew well on the medium and 157 blackened it. Other anaerobes, Enterobacteriaceae, and enterococci either failed to grow on BBE agar or did not produce the characteristic morphology and blackening associated with isolates of the B. fragilis group. In a clinical laboratory trial, 687 specimens from patients were inoculated onto BBE agar plates. The B. fragilis group was recovered from 81 (11.8%) of these specimens in 24 to 48 h. Use of BBE agar in the clinical laboratory enables earlier recovery and identification of this important pathogen.

Anaerobiosis↗
PubMed1978

Factors affecting production of catalase by Bacteroides.

Several variables affected the production of catalase by members of the "Bacteroides fragilis group" of anaerobic bacteria. Both media yielded higher catalase levels than the respective agar media. Addition of hemin to media after autoclave sterilization, rather than before, significantly increased production of catalase. Both of these variables could be related to the available hemin concentration present in the medium being tested. Significantly higher amounts of hemin were required for catalase production than were required for growth. For catalase production by B. fragilis ATCC 25285, 1 microgram of hemin per ml was required. Of the various media tested, the use of chopped meat broth resulted in the highest levels of catalase production (up to 50 to 60 U of catalase per mg of protein). Of the various species and DNA homology groups tested, strains of B. fragilis and Bacteroides distasonis were catalase positive. Strains of Bacteroides thetaiotaomicron, Bacteroides ovatus, and Bacteroides eggerthi possessed variable catalase activity. Bacteroides vulgatus, Bacteroides uniformis, and DNA homology groups "3452A" and "subsp. a" were catalase negative. A catalase well test, in which equal volumes of 3% H2O2 and chopped meat culture are mixed, is described and recommended for routine catalase tests.

Anaerobiosis↗
PubMed1979

Comparative serology of two clinical isolates of Bacteroides fragilis and Bacteroides thetaiotaomicron.

Antiserum prepared in rabbits against Bacteroides fragilis showed numerous bands when reacted with B. fragilis antigen in Ouchterlony plates. This antiserum also reacted with Bacteroides thetaiotaomicron and showed one band of apparent identity with B. fragilis. The band of identity common for both organisms was lost if the antigen was heated at 80 degrees C for 30 min. Antisera prepared against B. thetaiotaomicron did not react with B. fragilis.

Antigens, Bacterial↗
PubMed1975

Limb infections in infancy presenting with pseudoparalysis.

Three cases of pseudoparalysis in infancy are reported in which the appearance of localized infection in the affected limb was delayed for a variable period after the onset of pseudoparalysis. Blood cultures on admission in all cases were positive, and bacteraemia due to the three organisms isolated is discussed.

Ampicillin↗
PubMed1982

Pyogenic arthritis due to bacteroides complicating rheumatoid arthritis.

Two cases of anaerobic, pyogenic arthritis complicating rheumatoid disease due to bacteroides are described. The relative well-being of the patients and indolence of the affected joints are emphasised, and the effective role of metronidazole in the treatment of septic arthritis due to bacteroides is discussed.

Aged↗