Kenneth E. Aldridge

Association of Atopobium vaginae, a recently described metronidazole resistant anaerobe, with bacterial vaginosis

BackgroundBacterial vaginosis (BV) is a polymicrobial syndrome characterized by a change in vaginal flora away from predominantly Lactobacillus species. The cause of BV is unknown, but the condition has been implicated in diverse medical outcomes. The bacterium Atopobium vaginae has been recognized only recently. It is not readily identified by commercial diagnostic kits. Its clinical significance is unknown but it has recently been isolated from a tuboovarian abcess.MethodsNucleotide sequencing of PCR amplified 16S rRNA gene segments, that were separated into bands within lanes on polyacrylamide gels by denaturing gradient gel electrophoresis (DGGE), was used to examine bacterial vaginal flora in 46 patients clinically described as having normal (Lactobacillus spp. predominant; Nugent score ≤ 3) and abnormal flora (Nugent score ≥ 4). These women ranged in age from 14 to 48 and 82% were African American.ResultsThe DGGE banding patterns of normal and BV-positive patients were recognizably distinct. Those of normal patients contained 1 to 4 bands that were focused in the centre region of the gel lane, while those of BV positive patients contained bands that were not all focused in the center region of the gel lane. More detailed analysis of patterns revealed that bands identified as Atopobium vaginae were present in a majority (12/22) of BV positive patients, while corresponding bands were rare (2/24) in normal patients. (P < 0.001) Two A. vaginae isolates were cultivated from two patients whose DGGE analyses indicated the presence of this organism. Two A. vaginae 16S rRNA gene sequences were identified among the clinical isolates. The same two sequences were obtained from DGGE bands of the corresponding vaginal flora. The sequences differed by one nucleotide over the short (~300 bp) segment used for DGGE analysis and migrated to slightly different points in denaturing gradient gels. Both isolates were strict anaerobes and highly metronidazole resistant.ConclusionThe results suggest that A. vaginae may be an important component of the complex bacterial ecology that constitutes abnormal vaginal flora. This organism could play a role in treatment failure if further studies confirm it is consistently metronidozole resistant.

Multicenter Survey of the Changing In Vitro Antimicrobial Susceptibilities of Clinical Isolates of Bacteroides fragilis Group, Prevotella, Fusobacterium, Porphyromonas, and Peptostreptococcus Species

ABSTRACT In vitro surveys of antimicrobial resistance among clinically important anaerobes are an important source of information that can be used for clinical decisions in the choice of empiric antimicrobial therapy. This study surveyed the susceptibilities of 556 clinical anaerobic isolates from four large medical centers using a broth microdilution method. Piperacillin-tazobactam was the only antimicrobial agent to which all the isolates were susceptible. Similarly, imipenem, meropenem, and metronidazole were highly active (resistance, <0.5%), whereas the lowest susceptibility rates were noted for penicillin G, ciprofloxacin, and clindamycin. For most antibiotics, blood isolates were less susceptible than isolates from intra-abdominal, obstetric-gynecologic, and other sources. All isolates of the Bacteroides fragilis group were susceptible to piperacillin-tazobactam and metronidazole, while resistance to imipenem and meropenem was low (<2%). For these same isolates, resistance rates (intermediate and resistant MICs) to ampicillin-sulbactam, cefoxitin, trovafloxacin, and clindamycin were 11, 8, 7, and 29%, respectively. Among the individual species of the B. fragilis group, the highest resistance rates were noted among the following organism-drug combinations: for clindamycin,Bacteroides distasonis and Bacteroides ovatus; for cefoxitin, Bacteroides thetaiotaomicron, B. distasonis, and Bacteroides uniformis; for ampicillin-sulbactam,B. distasonis, B. ovatus, and B. uniformis; and for trovafloxacin, Bacteroides vulgatus. For the carbapenens, imipenem resistance was noted among B. fragilis and meropenem resistance was seen among B. fragilis, B. vulgatus, and B. uniformis. With few exceptions all antimicrobial agents were highly active against isolates of Prevotella, Fusobacterium, Porphyromonas, andPeptostreptococcus. These data further establish and confirm that clinically important anaerobes can vary widely in their antimicrobial susceptibilities. Fortunately most antimicrobial agents were active against the test isolates. However, concern is warranted for what appears to be a significant increases in resistance to ampicillin-sulbactam and clindamycin.

Infections related to the ingestion of seafood Part I: viral and bacterial infections

Foodborne diseases cause an estimated 76 million illnesses in the USA each year. Seafood is implicated in 10-19% of these illnesses. A causative agent can be traced in about 44% of seafood-related outbreaks, viruses accounting for around half of these illnesses. Although viruses are the most common cause of seafood-related infections, most hospitalisations and deaths are due to bacterial agents. A wide variety of viruses, bacteria, and parasites have been implicated in seafood-related outbreaks, which are reported worldwide. The factor most commonly associated with infection is consumption of raw or undercooked seafood. People with underlying disorders, particularly liver disease, are more susceptible to infection. The first part of this two-part review summarises the general incidence of seafood-related infections and discusses the common viral and bacterial causes of these infections. For each agent, the microbiology, epidemiology, mode of transmission, and treatment are discussed. In the May issue of the journal we will discuss parasites associated with seafood consumption, the safety of seafood, and the measures put in place in the USA to increase its safety.

Piperacillin/tazobactam (YTR 830) combination: Comparative antimicrobial activity against 5889 recent aerobic clinical isolates and 60 Bacteroides fragilis group strains

Piperacillin combined with tazobactam (formerly YTR 830) was tested at a ratio of 8:1 against 5889 aerobic isolates and 50 strains from the Bacteroides fragilis group. Imipenem was the most active agent tested against Enterobacteriaceae (99.3% at less than or equal to 4 micrograms/ml), ceftazidime was most effective against nonenteric Gram-negative bacilli (80.7% at less than or equal to 8 micrograms/ml), and piperacillin/tazobactam possessed a superior spectrum against Gram-positive cocci (92.2% at less than or equal to 16/2 micrograms/ml). Against all aerobic strains, piperacillin/tazobactam had a spectrum (90.3% at less than or equal to 16/2 micrograms/ml) comparable to imipenem (93.6% at less than or equal to 4 micrograms/ml) and was distinctly greater than that of ticarcillin/clavulanic acid (73.3% at less than or equal to 16/2 micrograms/ml) and ceftazidime (75.5% at less than or equal to 8 micrograms/ml). Against the B. fragilis group isolates, all piperacillin/tazobactam MICs were less than or equal to 64/8 micrograms/ml. This activity was superior to piperacillin alone (MIC:50, 8-64 micrograms/ml) and cefoxitin (MIC50, 4-64 micrograms/ml). Piperacillin/tazobactam appears to be a promising parenteral antimicrobial combination, with a spectrum effective against a wide variety of clinical pathogens.

Bacteremia due to Bacteroides fragilis group: distribution of species, beta-lactamase production, and antimicrobial susceptibility patterns.

ABSTRACT A retrospective analysis of susceptibility data on 542 blood isolates of the Bacteroides fragilis group tested from 1987 to 1999 by the same NCCLS-recommended broth microdilution method throughout is presented. Metronidazole, β-lactam-β-lactamase inhibitor combinations, carbapenems, and trovafloxacin were the most active agents (susceptibility of ≥93%). Among the cephalosporin-cephamycins, the order of activity was cefoxitin > ceftizoxime > cefotetan = cefotaxime = cefmetazole > ceftriaxone. All isolates were resistant to penicillin G, and 22% were resistant to clindamycin. The susceptibility rates to piperacillin-tazobactam, imipenem, and meropenem were affected least among isolates resistant to cefoxitin or clindamycin. Except for piperacillin-tazobactam, imipenem, and meropenem, the B. fragilis species was more susceptible than were the non-B. fragilis species. These data underscore the importance of susceptibility testing of the B. fragilis group and can serve as a guide in the choice of empirical antimicrobial therapy.

Comparison of the activities of penicillin G and new beta-lactam antibiotics against clinical isolates of Bacteroides species.

MICs were determined for 218 clinical isolates of Bacteroides by a broth microdilution method. Imipenem was the most active antibiotic tested. Azlocillin, mezlocillin, and cefoxitin had comparable activities, with resistance among members of the B. fragilis group and B. capillosus. Ceftizoxime was the most active cephalosporin tested. Members of the B. fragilis group showed high levels of resistance to cefotetan and ceftazidime. Resistance to penicillin G varied from 0 to 14%.

Antimicrobial activity of cefpirome an update compared to five third-generation cephalosporins against nearly 6000 recent clinical isolates from five medical centers☆

Cefpirome, cefotaxime, ceftazidime, cefoperazone, ceftizoxime, and ceftriaxone were tested against approximately 6000 fresh clinical isolates from five medical centers. For 3031 strains of Enterobacteriaceae tested, cefpirome consistently had the lowest MIC50s and lowest percentage of resistant strains. Cefpirome was also the most active agent against the 2138 Gram-positive cocci tested; Staphylococcus haemolyticus was uniformly resistant to all agents tested. Against 791 nonenteric Gram-negative bacilli, the activity of cefpirome was most comparable to that of cefoperazone and slightly less active than ceftazidime. Among the current third-generation cephalosporins, cefotaxime and cefoperazone emerged as having better overall balanced activity. Ceftazidime displayed poorest coverage against Enterobacteriaceae and Gram-positive organisms. Ceftizoxime also provided compromised coverage of staphylococci and nonenteric Gram-negative bacilli. Cefpirome remains as active as originally described in 1984 and possesses a slightly wider spectrum of activity against contemporary aerobic pathogens compared to currently marketed third-generation cephalosporins.

A five-year multicenter study of the susceptibility of the Bacteroides fragilis group isolates to cephalosporins, cephamins, penicillins, clindamycin, and metronidazole in the United States

Over 2800 clinical strains of the Bacteroides fragilis group were collected during a 5-year period from ten geographically separate sites and tested for their susceptibility to various antimicrobial agents using a broth microdilution method. Among the cephalosporins, ceftizoxime was the most active (13% resistance) and importantly exhibited relatively equal activity against both B. fragilis species and non-B. fragilis species. Cefotaxime exhibited similar activity with an overall resistance rate of 18%. Both ceftriaxone and cefoperazone were appreciably less active cephalosporins especially against non-B. fragilis species. With regard to cephamycins, cefoxitin (MIC90, 32 micrograms/ml) was more active than cefotetan (MIC90, > or = 256 micrograms/ml) and cefmetazole (MIC90, 64 micrograms/ml). Non-B. fragilis species were highly resistant to cefotetan and cefmetazole. Imipenem was highly active against all strains with the exception of four strains of B. fragilis. Ampicillin-sulbactam, amoxicillin-clavulanate, piperacillin-tazobactam, and cefoperazone-sulbactam were all highly active with resistance rates < 2%. No resistance was detected to metronidazole, whereas 14% of isolates were resistant to clindamycin. When compared with other studies, these findings underscore the wide variability in susceptibility patterns reported nationwide and the need to continue monitoring these patterns to aid in choosing the most active compounds for therapy.

Comparison of the in vitro activity of ciprofloxacin and 24 other antimicrobial agents against clinical strains of Chromobacterium violaceum

Eleven clinical strains of Chromobacterium violaceum were tested for their susceptibility to 25 antimicrobial agents. Ciprofloxacin was the most active of the compounds tested although norfloxacin and pefloxacin were highly active. No resistance was detected to mezlocillin, piperacillin, apalcillin, imipenem, and aztreonam while a single strain was resistant to ticarcillin. Among the cephalosporin/cephamycin group only cefotetan showed good in vitro activity. Gentamicin was more active than amikacin and tobramycin. Good in vitro activity was also noted for chloramphenicol, doxycycline, and trimethoprim-sulfamethoxazole while C. violaceum strains were highly resistant to rifampin and vancomycin. The bactericidal activity of selected agents was shown to be concentration dependent using time-kill kinetic studies. Addition of clavulanic acid did not increase the activity of ticarcillin and in one case was shown to induce beta-lactamase. High correlation was noted between the broth microdilution and disk diffusion susceptibility tests in predicting the susceptibility patterns of C. violaceum.

Comparison of the activities of coumermycin, ciprofloxacin, teicoplanin, and other non-beta-lactam antibiotics against clinical isolates of methicillin-resistant Staphylococcus aureus from various geographical locations.

One hundred clinical isolates of methicillin-resistant (Metr) Staphylococcus aureus from five different geographical origins were tested for their susceptibility to 12 non-beta-lactam antibiotics by the broth microdilution technique recommended by the National Committee for Clinical Laboratory Standards. Coumermycin and rifampin showed the highest activity with of MIC90s of 0.032 micrograms/ml for each compound. No resistance was found to coumermycin, whereas a single Metr S. aureus strain was found to be resistant to rifampin. Ciprofloxacin, vancomycin, and teicoplanin were the next most active compounds with MIC90s of 0.25, 0.5, and 0.5 micrograms/ml, respectively. Norfloxacin was less active, with an MIC90 of 1.0 micrograms/ml. Amikacin was the most active aminoglycoside tested, whereas high levels of resistance were found to tobramycin and gentamicin. Doxycycline and chloramphenicol showed variable results. Geographical variations in results were seen with doxycycline, chloramphenicol, and aminoglycosides. Kill-curve studies showed that coumermycin, ciprofloxacin, teicoplanin, vancomycin, and rifampin were highly bactericidal; more than 90% of the inoculum was killed within 8 h.