A comparison of aflatoxin B1-induced cytotoxicity, mutagenicity and prophage induction in Salmonella typhimurium mutagen tester strains TA1535, TA1538, TA98 and TA100.

Treatment of Ames mutagen tester strains with aflatoxin B1 (AFB1) and S9 mix results not only in the production of a potent mutagen, but induces a pathway that leads to the induction of prophages present in all Ames tester strains. Characterization of the prophage induction and mutagenic response following AFB1 treatment showed that plasmid pKM101 dramatically enhances mutagenesis, but suppressed prophage induction. Spontaneous release of phage by TA98 and TA100 was also lower than in TA1535 and TA1538. In addition to mutagenesis and prophage induction, survival of all 4 tester strains was quantitated after AFB1 treatment. The data show that the frameshift tester strains (TA1538 and TA98) are more sensitive to the bactericidal action of AFB1 than the base-pair tester strains (TA1535 and TA100), survival being significantly affected above 100 ng. One of several hypotheses examined was the difference in the number and types of prophages present in base-pair tester strains that are not detectable in the frame-shift tester strains. These data suggest that prophage induction can detect DNA damage that is non-mutagenic; and that it is important to characterize the lysogenic nature of the Ames strains since it may influence the observed histidine revertant rate and the survival of the tester strain.

Distribution and properties of the mannose-resistant hemagglutinin produced by Salmonella species.

Presence or absence of mannose-resistant hemagglutination (MRHA) or sheep erythrocytes by Salmonella species was found to be consistent within most serotypes tested and did not correlate with O-antigen. Broad-host-range serotypes including S. typhimurium and S. enteritidis produced MRHA while host-specific serotypes including S. typhi, S. dublin, and S. gallinarum were MRHA-negative. MRHA produced by S. typhimurium was soluble, heat-stable, and not inhibited by any of eleven carbohydrates tested. Further investigation of MRHA may provide insight into mechanisms of Salmonella pathogenesis.

Tn5030: a conjugative transposon conferring clindamycin resistance in Bacteroides species.

Several examples of conjugal transfer of antibiotic resistance determinants in the absence of plasmid DNA have been described in gram-positive species. Previous work had suggested that transfer of clindamycin resistance among the gram-negative anaerobic Bacteroides species might occur by a similar mechanism. We report here the characterization of Tn5030, a 45-kb +/- 5-kb chromosomal, mobile genetic element devoid of the detectable terminal repeats typically associated with gram-negative transposons. Tn5030 transfers clindamycin resistance (ermFU) and sequences homologous to tetF in the absence of detectable plasmid DNA. This element represents a novel mechanism of genetic exchange among gram-negative organisms.

Use of high-pressure liquid chromatography to determine plasma levels of metronidazole and metabolites after intravenous administration.

A rapid and sensitive high-pressure liquid chromatography assay for metronidazole and its two principle metabolites, 1-(2-hydroxyethyl-2-hydroxymethyl)-5-nitro-imidazole [hydroxy metabolite] and 1-acetic acid-2-methyl-5-metronidazole [acid metabolite], was developed. The retention times observed were 5.7, 3.3, and 4.5 min, respectively. A reverse-phase muC(18) Bondapak column using a solvent system of methanol, acetonitrile, and 0.005 M pH 4 potassium dihydrogen phosphate (4:3:93, vol/vol) was used to achieve separation of the three compounds. Patients receiving metronidazole therapy were given a loading dose of 13.6 mg of drug per kg intravenously over 1 h, followed by a maintenance dose of 1.43 mg/kg per h. The range of metronidazole concentrations observed was 6.8 to 47.5 mug/ml. These levels are well above the minimal inhibitory concentrations of most clinically significant anaerobic bacteria including Bacteroides fragilis. Little of the acid metabolite was observed in the plasma. The concentration of hydroxy metabolite ranged from 1.6 to 16 mug/ml. The latter may represent an additional source of antimicrobial activity since the hydroxy metabolite has approximately 30% the biological activity of metronidazole.

Genetic approaches to the study of oral microflora: a review.

As the study of oral microorganisms intensified almost 2 decades ago, the application of genetic techniques resulted in important contributions to the understanding of this clinically and ecologically important group of bacteria. The isolation and characterization of mutants of cariogenic streptococci helped to focus attention on traits that were important in colonization and virulence. Such classic genetic approaches gave way to molecular genetic techniques, including recombinant DNA methodology in the late 1970s. Gene cloning systems and methods to move DNA into cells have been developed for oral streptococci. Many streptococcal genes thought to be important in colonization and virulence have since been cloned and their nucleotide sequence determined. Mutant strains have been constructed using defective copies of cloned genes in order to create specific genetic lesions on the bacterial chromosome. By testing such mutants in animal models, a picture of the cellular and molecular basis of dental caries is beginning to emerge. These modern genetic methodologies also are being employed to develop novel and efficacious cell-free or whole cell vaccines against this infection. Genetic approaches and analyses are now being used to dissect microorganisms important in periodontal disease as well. Such systems should be able to exploit advances made in genetically manipulating related anaerobes, such as the intestinal Bacteroides. Gene cloning techniques in oral anaerobes, Actinomyces and Actinobacillus, are already beginning to pay dividends in helping understand gene structure and expression. Additional effort is needed to develop facile systems for genetic manipulation of these important groups of microorganisms.