Susceptibility of Actinobacillus actinomycetemcomitans to six antibiotics decreases as biofilm matures

OBJECTIVES: Actinobacillus actinomycetemcomitans is a major causative agent of chronic and aggressive periodontitis. Freshly isolated strains of A. actinomycetemcomitans display rough-type colonies and initiate biofilm formation on glass surfaces. The purpose of this study was to determine the antibiotic susceptibility of A. actinomycetemcomitans biofilm during different phases of maturation. METHODS: Using 96-well microtitre plates, we determined the antibiotic susceptibility of rough-type strain 310a to concentrations from 0.1 to 10 mg/L each of erythromycin, ofloxacin, ampicillin, cefalexin, tetracycline and minocycline during biofilm formation. Antibiotics were added at the start of the culture (early phase) and after 24 h of cultivation (mature phase). RESULTS: Adding 10 mg/L of ampicillin, 10 mg/L of cefalexin, 0.1 or 1 mg/L of tetracycline, or 0.1 mg/L of minocycline significantly inhibited 310a biofilm formation in the early phase, but not in the mature phase. Although adding 10 mg/L of erythromycin, tetracycline or minocycline reduced biofilm development in the early phase, it enhanced 310a biofilm development in the mature phase. Ofloxacin exerted a strong inhibitory effect in both the early and mature phases of biofilm formation throughout all experiments. CONCLUSIONS: The present study demonstrated that the susceptibility of A. actinomycetemcomitans to many antibiotics decreased after biofilm maturation.     

Effect of clindamycin, erythromycin, lincomycin, and tetracycline on growth and extracellular lipase production by propionibacteria in vitro.

Two propionibacteria identified as Propionibacterium acnes and Propionibacterium granulosum were grown anaerobically in the presence of growth subinhibitory concentrations (0.25 and 0.5 minimal inhibitory concentrations) of clindamycin, erythromycin, lincomycin, and tetracycline. Viable counts and assays of extracellular lipase were performed on samples taken at 24-h intervals over a 96-h period. The results showed that lincomycin and clindamycin could inhibit the production of the enzyme by both strains with little effect on their growth rates. Tetracycline caused inhibition of lipase production by P. granulosum only. Although production of the enzyme by P. acnes was delayed in the presence of tetracycline, the final titer was the same as the control. Erythromycin had little effect on growth and enzyme production of either strain. It is possible, therefore, that certain antibiotics used in acne therapy may act not only as bactericidal agents but also as inhibitors of enzyme production under non-growth-limiting conditions.     

Evaluation of the Performance Parameters of a Prediluted, Quantitative Antibiotic Susceptibility Test Device

A multiwelled plastic strip containing prediluted antibiotics was tested. Antibiotics available at the time of testing included ampicillin, penicillin, methicillin, erythromycin, clindamycin, tetracycline, and minocycline. The effects of inoculum size, inoculum volume, temperature, and time of incubation were determined. A limited clinical evaluation using laboratory strains of selected bacteria proved this product to be a rapid, economical, and reliable method for the determination of the minimal inhibitory concentration of antibiotics.     

In vitro antimicrobial susceptibility of Actinobacillus actinomycetemcomitans

The agar dilution technique was used for determination of the antibiotic susceptibilities of 57 oral isolates and 2 nonoral isolates of Actinobacillus actinomycetemcomitans. Tetracycline, minocycline, and chloramphenicol inhibited more than 96% of the strains tested at a concentration of less than or equal to 2 micrograms/ml; 89% of the strains were inhibited by 2 micrograms of carbenicillin per ml. The other antimicrobial agents tested were less active. Approximately 10% of the A. actinomycetemcomitans strains were resistant to ampicillin, erythromycin, and penicillin G at concentrations of 32 to 64 micrograms/ml. These data suggest that tetracycline and minocycline may be valuable drugs in the treatment of A. actinomycetemcomitans infections.     

Isolation and Characterization of Multiply Antibiotic-Resistant Clostridium perfringens Strains from Porcine Feces

Multiply antibiotic-resistant strains of Clostridium perfringens were isolated from porcine feces. Strains that were resistant to tetracycline, erythromycin, clindamycin, and lincomycin were isolated, but no penicillin- or chloramphenicol-resistant strains were obtained. Typical minimal inhibitory concentrations for resistant strains were 16 to 64 mug of tetracycline per ml, 64 to >128 mug of erythromycin per ml, >/=128 mug of lincomycin per ml, and 16 to 128 mug of clindamycin per ml. Resistance to erythromycin was always associated with resistance to lincomycin and clindamycin. Minimal inhibitory concentrations were determined for 258 strains from six farms that used antibiotics in their feeds and 240 strains from five farms that did not use antibiotics. The results show that 77.9 and 22.7% of the strains from the former farms were resistant to tetracycline and erythromycin-clindamycin-lincomycin, respectively. The comparable data from the latter farms were 25.0 and 0.8%, respectively. Agarose gel electrophoresis failed to reveal a plasmid band that was common to the resistant strains but absent in the susceptible strains. Attempts to transfer tetracycline, erythromycin, and clindamycin resistance from one strain, CW459, were not successful. Antibiotic-susceptible mutants were not isolated from this strain, despite the use of a variety of curing agents.     

Antibiotic susceptibility profile of bifidobacteria as affected by oxgall, acid, and hydrogen peroxide stress

The effects of acid, oxgall, and H(2)O(2) on susceptibilities to antibiotics and nisin were examined for 13 strains of bifidobacteria. Susceptibilities to ampicillin, cloxacillin, penicillin, vancomycin, kanamycin, neomycin, paramomycin, streptomycin, chloramphenicol, erythromycin, tetracycline, and nisin A were assayed by a microdilution broth method. Acid-, oxgall- and H(2)O(2)-stressed variants were produced and assayed. Exposure to a pH of 2.0 for 60 min reduced susceptibilities to cloxacillin and nisin A but increased susceptibilities to ampicillin, vancomycin, aminoglycosides, chloramphenicol, and erythromycin in a strain-dependent manner. Exposure to oxgall (0.3%) for 90 min increased susceptibilities to cell wall-directed antibiotics and aminoglycosides but increased resistances to tetracycline and nisin A. Oxidative stress increased the susceptibilities of 70% of the strains to ampicillin and chloramphenicol, of 50% of the strains to cloxacillin and tetracycline, and of 40% of the strains to erythromycin but did not affect susceptibilities to vancomycin, kanamycin, and nisin A. This study shows that exposure of bifidobacteria to stressful conditions resembling those in the gastrointestinal tract may substantially modify their susceptibilities to antibiotics and may thus affect their probiotic capacities, especially when they are used for the management of intestinal infections and antibiotic-associated diarrhea.     

Antimicrobial susceptibility of Haemophilus influenzae

Selected clinical isolates of Haemophilus influenzae were tested for their susceptibility to seven antibiotics by a microtiter broth dilution and an agar diffusion method. Eleven of 40 strains tested were resistant to ampicillin, the drug of choice, by both methods. All the strains tested were susceptible to chloramphenicol, and all but one were susceptible to tetracycline. Of the other four antibiotics tested, the ranges of minimal inhibitory concentrations (MICs) were: 0.5 to 4 mug of gentamicin per ml, 0.5 to 4 mug of erythromycin per ml, and 2 to 16 mug of clindamycin per ml. MICs of ampicillin for both the susceptible and resistant strains were markedly affected by inoculum concentration. The ampicillin MICs of the resistant strains were also affected by the time of incubation.     

Effect of ampicillin, cefmetazole and minocycline on the adherence of Branhamella catarrhalis to pharyngeal epithelial cells

Using pharyngeal epithelial cells from a healthy adult and eight strains of Branhamella catarrhalis (B. catarrhalis) isolated from eight patients with respiratory infection the effect of subminimal inhibitory concentrations of cefmetazole, ampicillin and minocycline on adherence was examined. Cefmetazole-treated bacterial attachment (44 +/- 28; mean +/- S.D.) decreased significantly (p less than 0.05) compared to the control (84 +/- 27). Statistically no significant difference in adherence was found between ampicillin-treated bacteria (63 +/- 36) and the control (95 +/- 40) or minocycline-treated bacteria (91 +/- 39) and the control (109 +/- 40). Large bacteria was observed after cefmetazole and ampicillin treatment. In addition to diplococci, tetrads were observed after cefmetazole treatment. Significant correlation between the MICs and adherence ability was not found. The results suggests that these three antibiotics were not responsible for the increase in B. catarrhalis infection by increasing adherence ability.     

Antimicrobial Susceptibility of Haemophilus parainfluenzae

Fifty random clinical isolates of Haemophilus parainfluenzae were tested for their susceptibility to 10 antibiotics by a microtiter broth dilution method. Three of the strains tested were resistant to ampicillin, whereas eight were resistant to tetracycline. All strains tested were susceptible to chloramphenicol, kanamycin, gentamicin, cephalothin, and colistin. The ranges of minimal inhibitory concentrations for the three remaining antibiotics were: 0.5 to >/=128 mug of penicillin G per ml, 0.03 to 4 mug of carbenicillin per ml, and 1 to 16 mug of erythromycin per ml. Elevated minimal inhibitory concentrations for penicillin and carbenicillin were noted for the three ampicillin-resistant strains. Tests for beta-lactamase production demonstrated the presence of this enzyme in each of the three ampicillin-resistant strains.     

Antimicrobial Susceptibility of Propionibacterium acnes and Related Microbial Species

The minimal inhibitory concentrations of 32 antimicrobial agents were established for 73 strains of Propionibacterium acnes and four related species (P. granulosum, P. avidum, Corynebacterium minutissimum, and C. parvum). Most strains showed good susceptibility to those agents usually considered active against gram-positive organisms. With the exception of C. minutissimum, the strains tested revealed more or less identical susceptibility ranges. The lowest minimal inhibitory concentrations were observed with benzylpenicillin, ampicillin, cephalothin, rifampin, erythromycin, clindamycin, and minocycline. C. minutissimum was more susceptible to gentamicin, sisomicin, tobramycin, and fusidic acid but more resistant to most other drugs than were the other species examined.     

In vitro susceptibilities of Ehrlichia risticii to eight antibiotics.

Inhibition of the proliferation of Ehrlichia risticii cultured in murine macrophage P388D1 cells by eight antibiotics was evaluated by indirect fluorescent-antibody staining with an antiserum specific to E. risticii. There was a negative correlation between the percentage of infected cells and the log10 of the concentrations of all antibiotics examined. The ranks of the antibiotics in the order of 50% inhibitory concentrations (on a microgram-per-milliliter basis) after 48 h of exposure were as follows: demeclocycline, doxycycline, and oxytetracycline less than minocycline less than rifampin less than tetracycline less than erythromycin and nalidixic acid. When the antibiotics were removed after 48 h of incubation, continuous inhibition of proliferation was evident at 72 h. At 96 h regrowth of the organisms occurred in most of the cultures. The rate of regrowth was the highest with nalidixic acid, followed by erythromycin, at all concentrations of the antibiotic tested. Regrowth was observed with less than 0.1 microgram of minocycline per ml and less than 0.01 microgram of oxytetracycline, tetracycline, and doxycycline per ml. With more than 0.01 microgram of demeclocycline per ml, however, the inhibition persisted for up to 72 h after removal of the antibiotic. These results indicate that demeclocycline was slightly more effective than doxycycline, oxytetracycline, and minocycline in eliminating E. risticii in macrophages in vitro, whereas tetracycline and rifampin were less effective. Nalidixic acid and erythromycin were ineffective.     

空肠弯曲菌耐药谱特征分析

目的 分析十几年间我国空肠弯曲菌临床分离株对10种抗生素耐药谱特征,了解我国空肠弯曲菌耐药的变迁趋势。 方法 采用世界卫生组织(WHO)全球食源性病原菌感染网络(GFN)推荐的弯曲菌琼脂稀释法,测定1995年至今分离的116株空肠弯曲菌对6类10种抗生素的最小抑菌浓度(MIC)。 结果 经对实验结果整体分析,甲硝唑的总体耐药率最高为97.4%(113/116),四环素为82.8%(96/116),环丙沙星为80.2%(93/116),萘啶酸为79.3%(92/116),左氧氟沙星和氨苄西林耐药率相同,为40.5%(47/116),氯霉素为18.1%(21/116),庆大霉素为8.6%(10/116),链霉素为4.3%(5/116),最低为红霉素0(0/116)。随着时间的推进,萘啶酸、环丙沙星、左氧氟沙星和氨苄西林的MIC有明显增高趋势;四环素、红霉素、庆大霉素、氯霉素和甲硝唑的MIC值变化不明显;链霉素的MIC值变化有下降的趋势。6.1%的菌株出现了8种抗生素多重耐药的结果,且菌株均出现在2010年后。经统计学分析,萘啶酸、环丙沙星、链霉素、庆大霉素、氯霉素和氨苄西林6种抗生素在2001年前、2001-2005年、2006-2010年和2010年后4个时间段中耐药率差异有统计学意义。 结论 空肠弯曲菌对红霉素、庆大霉素以及链霉素3种抗生素依旧保持了较高的敏感性,对萘啶酸、环丙沙星、左氧氟沙星、四环素、甲硝唑以及氨苄西林6种抗生素产生了较大程度的耐药。     

Antibiotic Susceptibility of Streptococcus bovis and Other Group D Streptococci Causing Endocarditis

Seventy-four strains of Streptococcus bovis and 35 strains of enterococci (Streptococcus faecalis and its varieties, Streptococcus faecium and Streptococcus durans), most of which were isolated from patients with endocarditis, were tested for their susceptibility to penicillin, ampicillin, erythromycin, cephalothin, vancomycin, methicillin, tetracycline, chloramphenicol, kanamycin, streptomycin, and gentamicin. Minimal inhibitory concentrations (MIC) and minimal bactericidal concentrations (MBC) were determined by a microtiter broth dilution technique. All of these organisms are group D streptococci, but the S. bovis strains are not enterococci. On the basis of both MIC and MBC, the S. bovis strains were much more susceptibile in general to antibiotics then were the enterococcal strains. For the S. bovis strains, the lowest MICs were obtained with penicillin, ampicillin, and erythromycin, and the lowest MBCs with penicillin and ampicillin. Although these antibiotics were also the most active against the enterococci, the MICs and MBCs were much higher than obtained with the S. bovis strains. Gentamicin was the most active aminoglycoside. On the basis of in vitro susceptibility results, the S. bovis strains resemble the viridans streptococci rather than enterococci.     

Antibiotic susceptibilities of human isolates of Pasteurella multocida.

Seventeen human strains of Pasteurella multocida, biochemically similar to, if not identical with, isolates of animal origins, were tested for susceptibility to 16 antimicrobial agents utilizing a microtiter broth dilution technique. Ten of these isolates were also tested against 11 antibiotics by disk diffusion. The most active drugs with respect to the median minimal inhibitory concentration (micrograms per milliliter) were tetracycline (0.09), penicillin G (0.78), ampicillin (0.78), carbenicillin (1.56), cephalothin (1.56), and chloramphenicol (1.56). With the exception of tetracycline, the median minimal inhibitory concentration and minimal bactericidal concentration values were equal or differed by no more than a factor of two. The semisynthetic penicillins clindamycin, erythromycin, and aminoglycosides had relatively low activities, suggesting that these agents would be poor choices for the treatment of P. multocida infections.     

Effects of subminimal inhibitory concentrations of Erythromycin, Clindamycin, and Pristinamycin on the penicillinase production of Staphlyococcus aureus

The influence of subminimal inhibitory concentrations of erythromycin, clindamycin, and pristinamycin on the penicillinase production of Staphylococcus aureus was tested in 12 strains. Of the 36 experiments performed, 16 (44%) showed a lack of influence, 10 (28%) displayed an increase, and 10 revealed a decrease in penicillinase activity. The maximal effect produced was generally induced by concentrations ranging from 1/4 to 1/32 the minimal inhibitory concentration, irrespective of the susceptibility of the strain to the drug. In spite of the fact that the drugs are closely related, they sometimes produced opposite effects on the same strain.     

Susceptibility of Related Vibrios and Vibrio Fetus to Twelve Antibiotics

One hundred fourteen strains of related vibrio and seven strains of Vibrio fetus, all isolated from humans, were tested for susceptibility in vitro to 12 antibiotics. Mueller-Hinton agar containing 14 dilutions of the antibiotics was inoculated with undiluted overnight cultures. Gentamicin and erythromycin were the most active drugs against related vibrios and Vibrio fetus; lower activity was noted with chloramphenicol, streptomycin, and tetracycline; neomycin and kanamycin were even less active. The susceptibility to ampicillin was variable. More than 90% of the strains were resistant to cephalothin.     

In vitro susceptibility of Ureaplasma urealyticum clinical isolates to new macrolides.

Nine antimicrobial agents, the new macrolides, rokitamycin and midecamycin acetate, and seven other antibiotics, tetracycline, minocycline, doxycycline, josamycin, erythromycin, spiramycin, and norfloxacin, were studied for their antimicrobial activity against 100 strains of Ureaplasma urealyticum, using a microtiter broth dilution technique. The new macrolides, rokitamycin and midecamycin acetate, had the highest activity, with the MIC against 90% of isolates tested (MIC90) being less than or equal to 0.05 microgram/ml. MICs90 of erythromycin, josamycin, doxycycline, minocycline and tetracycline ranged from 0.1 to 0.78 micrograms/ml. Norfloxacin was least active, with a MIC90 of 12.5 micrograms/ml. Five of 100 strains tested were resistant (MIC greater than or equal to 12.5 micrograms/ml) to tetracycline, and two were resistant to minocycline and doxycycline; all of these were susceptible to rokitamycin and midecamycin acetate.     

Bacteriostatic and Bactericidal Activities of Various Antibiotics Against Bacteroides fragilis

The minimal inhibitory concentrations and minimal bactericidal concentrations of seven antibiotics were determined by broth dilution tests for 50 clinical isolates of Bacteroides fragilis. Chloramphenicol, erythromycin, lincomycin, clindamycin, and rifampin inhibited 80 to 100% of the strains at concentrations attainable in the serum. Penicillin and tetracycline were less effective, inhibiting 4 and 40% of the strains, respectively, at concentrations attainable in the serum. The strains exhibited a bimodal distribution with tetracycline but a very narrow pattern of susceptibility with chloramphenicol. Bactericidal concentrations were 4- to 128- fold higher than minimal inhibitory concentrations for all antibiotics; only clindamycin showed bactericidal activity at levels attainable in serum. Clindamycin was significantly more effective than lincomycin as determined by tests of inhibitory and bactericidal activity. The distinct patterns of susceptibility of B. fragilis may be used for the preliminary selection of antimicrobial therapy.     

Susceptibility of Streptococcus mutans to Antimicrobial Agents

Fifty strains of Streptococcus mutans, including defined strains and clinical isolates, were tested for susceptibility to 20 different antimicrobial agents. Minimal inhibitory concentrations were determined by a liquid microtiter procedure. Antibiotics that were most effective in concentrations below 0.1 μg/ml included penicillin, ampicillin, erythromycin, cephalothin, and methicillin. Antibiotics effective in concentrations between 0.1 μg and 10 μg/ml included rifampin, lincomycin, thiostrepton, spiromycin, vancomycin, streptolydigan, novobiocin, tetracycline, chloramphenicol, spectinomycin, and gentamicin. Antibiotics effective at higher concentrations ranging from 10 μg/ml to 400 μg/ml included the aminoglycosides kanamycin, neomycin, streptomycin, and kasugamycin. Although most antibiotics exhibited inhibitory effects in a narrow range of concentrations, antibiotics such as tetracycline, thiostrepton, and spiromycin had a 1,000-fold range from the lowest to highest concentrations required for growth inhibition.     

Susceptibility of Pneumococci and Haemophilus influenzae to Antibacterial Agents

Strains of Diplococcus pneumoniae and Haemophilus influenzae were tested for susceptibility to numerous antibiotics by a twofold agar dilution method using an inocula replicator. Undiluted, fully grown broth cultures were used as inocula for both species, and cultures of pneumococci diluted 1:1,000 were also tested. The antibiotics included most of those in common use in the United States as well as some chemical modifications recently approved and others that are under investigation. The most striking aspect of the results was the marked susceptibility of the pneumococci to all the antibiotics tested except the polymyxins and most of the aminoglycoside antibiotics, although some new aminoglycosides were active in quite low concentrations. Some of the strains of pneumococci were of decreased susceptibility to penicillin G (minimal inhibitory concentrations, 0.2 to 0.4 mug/ml), but none were tetracycline resistant, although such strains had been reported previously from this laboratory. The strains of H. influenzae, which were all serologically nontypable, exhibited different patterns of susceptibility to the groups of antibiotics and to the individual chemically related ones. None of these strains (isolated early in 1972) were ampicillin resistant. The most active agents against H. influenzae were: carbenicillin and ampicillin, analogues related to each of them, rifampin, chloramphenicol, and the polymyxins. However, the tetracycline analogues other than tetracycline, some aminoglycosides, notably tobramycin, kanamycin, gentamicin, and verdamicin, erythromycin, and some new lincomycin analogues were also active in low concentrations. Trimethoprim alone was highly active, and in combination with sulfamethoxazole it was even more active and synergistic against strains of both D. pneumoniae and H. influenzae.