Carol J. Shanholtzer

Ten years of prospective Clostridium difficile-associated disease surveillance and treatment at the Minneapolis VA Medical Center, 1982-1991.

OBJECTIVES To understand the epidemiology, risks, and management of Clostridium difficile-associated disease (CDAD) and to establish and evaluate reliable methods of surveillance. DESIGN Case finding was done by daily ward and laboratory rounds. The criteria for CDAD diagnosis were: at least four unformed stools per day for 2 days and a positive culture or cytotoxin for C difficile, or positive endoscopy or autopsy for pseudomembranes. SETTING The surveillance covered all patients from 1982 through 1991 in the 820-bed Minneapolis Veterans Affairs Medical Center. PARTICIPANTS The criteria were met by 908 patients. Medical service patients numbered 488; surgical patients, 420. Frequencies ranged from a high of 149 cases in 1982 to a low of 50 cases in 1989. RESULTS Stool specimens were obtained on 898 (99%) of the 908 CDAD patients. Stools were culture-positive in 864 (96%) of 898, cytotoxin-positive in 569 (63%) of 898. Endoscopy was performed on 196 (22%) of the 908 patients, and 80 (41%) of 196 patients had pseudomembranes. Ten (1%) of the 908 patients were diagnosed by endoscopy without a stool specimen, or at autopsy. No treatment was needed for 135 (15%) of the 908 CDAD patients, and 19 (2%) of the 908 died before treatment was started. Oral metronidazole was the treatment for 632 (70%) of 908 patients (1% intolerance, 2% failure, 7% relapse) and oral vancomycin was given to 122 (13%) of 908 patients (1% intolerance, 1% failure, 10% relapse). Twelve patients had pseudomembranous colitis at autopsy, and it was the primary cause of death in 5 (0.6%) of 908. CONCLUSIONS CDAD usually responds to oral metronidazole or vancomycin but is nonetheless responsible for a high morbidity and occasional mortality in patients even when the diagnosis and treatment are pursued aggressively.

Aminoglycoside resistance and aminoglycoside usage: ten years of experience in one hospital.

For 10 years the 700-bed Minneapolis Veterans Affairs Medical Center has conducted a policy of carefully controlled aminoglycoside usage and monitoring of resistance of over 25,000 aerobic and facultative gram-negative bacillary isolates to the aminoglycosides. On two occasions during the 1980s, our experience of introducing amikacin at a high level of usage was associated with a significant reduction in resistance to gentamicin and tobramycin among gram-negative bacilli. Rapid reintroduction of gentamicin usage in 1982 after the first amikacin period was associated with a significant and rapid increase in gentamicin and tobramycin resistance. However, in 1986, gentamicin was again reintroduced to this institution at an initially modest level, and the percentage of usage of gentamicin was gradually increased over a 15-month period without a significant change in resistance to gentamicin, tobramycin, or amikacin while maintaining an overall 68% gentamicin usage and 30% amikacin usage. Aminoglycoside usage (measured as patient days) rose steadily from under 2,000 patient days per quarter in 1980 and 1981 to over 3,000 days per quarter in 1985. Since 1985, usage has declined to under 2,500 patient days per quarter in 1990. This usage rise and fall occurred during a steadily declining daily patient census that was 590 in 1980 and 465 in 1989. A move to a new hospital building in June 1988 was associated with an additional significant decline in resistance to all aminoglycosides (P less than 0.05), continuing a trend that was evident for the year preceding the move. Resistance to aminoglycoside antibiotics is now at the lowest level in 10 years at this institution, with only one gram-negative organism, Pseudomonas aeruginosa, that exhibits more than 5% resistance to gentamicin and no gram-negative species that are more than 5% resistant to amikacin and tobramycin.

Pseudomonas infection of the biliary system resulting from use of a contaminated endoscope

Pseudomonas aeruginosa was present in bile cultures from 10 patients who had undergone previous endoscopic retrograde cholangiopancreatography in 1984. After environmental cultures and review of instrument disinfection, we traced the infections to a single endoscope contaminated with P. aeruginosa, serotype 10. Although the instrument had been cleaned repeatedly with an automatic endoscope cleaning machine, P. aeruginosa survived on residual moisture left in the channels of the endoscope. Contamination ended only after we began to manually suction alcohol through the endoscope before air drying. In 5 of 10 patients, P. aeruginosa caused clinical infections including gangrenous cholecystitis, abscesses, and death. We could identify no factor that distinguished symptomatic from asymptomatic patients. In asymptomatic patients, P. aeruginosa was recovered from gallbladder bile up to 2 mo after endoscopic retrograde cholangiopancreatography. As this P. aeruginosa epidemic was discovered retrospectively because we monitor bile cultures, we advocate this practice as part of endoscopic retrograde cholangiopancreatography procedures.

Tests for bactericidal effects of antimicrobial agents: technical performance and clinical relevance.

Bactericidal testing has been used for several decades as a guide for antimicrobial therapy of serious infections. Such testing is most frequently performed when bactericidal antimicrobial agent therapy is considered necessary (such as when treating infectious endocarditis or infection in an immunocompromised host). It has also been used to ensure that the infecting organism is killed by (not tolerant to) usually bactericidal compounds. However, few data are available to support the role of such tests in direct patient care. Several important variables affect the reproducibility of the test results; however, proposed reference methods are now available for performing the MBC test. With minor modifications, these can provide a standardized approach for laboratories that need to perform them. Currently, little evidence is available to support the routine use of such testing for the care of individual patients. However, testing of new (investigational) antimicrobial agents can be beneficial in determining their potential to provide bactericidal antimicrobial activity during clinical use. New methods to assess bactericidal activity are being developed, but as yet none have been rigorously tested in patient care settings; further, for most of these methods, little information is available as to which technical parameters affect their results. In clinical laboratories, all bactericidal tests must be performed with rigorously standardized techniques and adequate controls, bearing in mind the limitations of the currently available test procedures.

Results of a prospective, 18-month clinical evaluation of culture, cytotoxin testing, and culturette brand (CDT) latex testing in the diagnosis of Clostridium difficile-associated diarrhea

An 18-mo evaluation of culture, cytotoxin, and latex testing for Clostridium difficile was performed between July 1, 1985, and December 31, 1986, on 1,536 specimens from 1,406 patients during evaluation of diarrhea. All cases with at least one test positive were investigated for clinical status. There were 144 Clostridium difficile-associated diarrhea (CAD) patients; 139 (97%) were positive by culture, 96 (67%) by cytotoxin, and 98 (68%) by latex testing. In the 1,262 non-CAD patients with diarrheal stool, 89 (7.1%) were positive by culture, 18 (1.4%) by cytotoxin, and 68 (5.4%) by the latex test. No CAD patient was positive by cytotoxin testing only, and two were positive by latex testing only. The culture and cytotoxin positivity were similar to our previous reports of 90-97% and 70-73%, respectively. Latex sensitivity (68%) was comparable to that of cytotoxin testing in this large group of patients (p greater than 0.5). Overall, in the 1,262 patients without clinical evidence of Clostridium difficile disease, positive tests by latex testing (5.4%) were intermediate between those of culture (7.1%, p less than 0.1) and cytotoxin (1.4%, p less than 0.001).

MBCs for Staphylococcus aureus as determined by macrodilution and microdilution techniques.

MBC testing of clindamycin, methicillin, cephalothin, gentamicin, and vancomycin with 67 clinical isolates of Staphylococcus aureus was examined by both standard macrodilution tubes and commercial microdilution trays. Standard macrodilution failed to give reproducible (99.9% killing) MBC results, even when a strictly defined protocol was followed. Continuous shaking during incubation resulted in regrowth of more colonies than did stationary incubation. Vortexing of incubated tubes before subculture resulted in regrowth of more colonies than did careful transfer of the contents to sterile tubes before vortexing and subculture. No significant difference in MBCs was demonstrated by the use of log-phase versus stationary-phase inocula. Use of the multiprong inoculator for subculture from commercial microdilution trays was unsatisfactory because, although antibiotics evaluated were inactivated by subculture to a pH 5.5 agar plate coated with a beta-lactamase solution, the volume of broth transferred by the prongs was small and inconsistent, ranging from 0 to 3 microliter. Subcultures of commercial microdilution panels with a 1-microliter loop, 10-microliter pipette, and 100-microliter pipette were also evaluated. Results of MBC testing were most reproducible when the entire 100-microliter volume was aspirated from commercial microdilution wells after stirring and the contents of each well were spread over a separate sheep blood agar plate. Images

In vitro activity of efrotomycin, ciprofloxacin, and six other antimicrobials against Clostridium difficile

The susceptibility of 69 clinical isolates of Clostridium difficile from the Minneapolis Veterans Administration Medical Center and 29 C. difficile strains from other hospitals to efrotomycin, ciprofloxacin, and six other antimicrobials was tested in vitro by agar dilution. Ciprofloxin (MIC50 and MIC90 = 8 mcg/ml) was only moderately active whereas efrotomycin (MIC50 = 0.125, MIC90 = 0.25 mcg/ml) was highly active against C. difficile.

Activity of twenty-one antimicrobial agents including l-ofloxacin against quinolone-sensitive and -resistant, and methicillin-sensitive and -resistant Staphylococcus aureus.

There is a need to identify alternative agents to vancomycin for the treatment of infections with methicillin-resistant Staphylococcus aureus (MRSA). One candidate is the l isomer of ofloxacin (DR-3355). We tested 520 frozen MRSA isolates, 248 fresh MRSA isolates, and 375 fresh methicillin-susceptible S. aureus (MSSA) isolates from Minnesota, and 600 clinical isolates of S. aureus (150 MRSA and 450 MSSA) from Illinois. Over 90% of the MRSA strains were resistant to 32 micrograms/ml of oxacillin. Of the 520 frozen MRSA, 24% were susceptible to < or = 2 micrograms/ml ofloxacin, and an additional 74% were susceptible to ofloxacin between 8 and 16 micrograms/ml. More than 98% of all strains were susceptible to < or = 16 micrograms/ml ofloxacin or l-ofloxacin. All the quinolones had a bimodal distribution of in vitro activity, but for only ofloxacin and l-ofloxacin was activity confined to a very narrow range.

Selenomonas bacteraemia — Case report and review of the literature

Selenomonas species are crescent shaped Gram-negative bacilli with a characteristic tuft of flagella located on the concave surface. They are normally found in human gingiva or the rumen of herbivores. The first case of Selenomonas bacteraemia to be reported in a patient immunocompromised by malignant disease is described and the two previously reported cases of Selenomonas bacteraemia as reviewed. The importance of careful anaerobic culturing to recover the organism and special diagnostic techniques to classify the bacteria as Selenomonas species are emphasised. These organisms may cause serious human disease including bacteraemia.

Bactericidal action of gentamicin against enterococci that are sensitive, or exhibit low- or high-level resistance to gentamicin

Treatment of serious enterococcal infection involves the use of penicillin-aminoglycoside combination therapy if the aminoglycoside minimum inhibitory concentration (MIC) is < or = 2000 micrograms/ml, and the organism is susceptible to penicillin or ampicillin. We evaluated killing of 15 enterococci that differ in their susceptibility to gentamicin using time-kill studies at different gentamicin concentrations. Sensitive strains had a uniform population killed by gentamicin concentrations equal to or above the MIC. Low-level resistant strains (MIC > or = 8 but < or = 2000 micrograms/ml of gentamicin) had a diverse population with large numbers of cells killed at one-half the MIC, while the highly resistant strains (MIC > 2000 micrograms/ml) showed no killing by any concentration of gentamicin.