Inaccuracy of the disk diffusion method compared with the agar dilution method for susceptibility testing of Campylobacter spp.

The agar dilution method has been standardized by the CLSI for the susceptibility testing of Campylobacter species, and according to these standards, the disk diffusion method should be used only in screening for macrolide and ciprofloxacin resistance. Nevertheless, the disk diffusion test is currently widely used, since it is easy to perform in clinical microbiology laboratories. In this study, the disk diffusion method was compared to the agar dilution method by analyzing the in vitro activities of seven antimicrobial agents against 174 Campylobacter strains collected in Finland between 2003 and 2008. Recommendations of the CLSI were followed using Mueller-Hinton agar plates with 5% of sheep blood. For each strain, the disk diffusion tests were performed two to four times. Of the 33 erythromycin-resistant strains (MIC, ≥16 µg/ml), 24 (73%) constantly showed a 6-mm erythromycin inhibition zone (i.e., no inhibition), while for seven strains the inhibition zone varied from 6 to 44 mm in repeated measurements. Among the 141 erythromycin-susceptible strains (MIC, <16 µg/ml), erythromycin inhibition zones varied between 6 and 61 mm. Of the 87 ciprofloxacin-resistant strains, 47 (54%) showed 6-mm inhibition zones, while 40 strains showed inhibition zones between 6 and 60 mm. Significant differences between the repetitions were observed in the disk diffusion for all antimicrobial agents and all strains except for the macrolide-resistant strains regarding the macrolides. For 17 (10%) strains, the variation in repeated measurements was substantial. These results show that the disk diffusion method may not be a reliable tool for the susceptibility testing of Campylobacter spp. Further studies are needed to assess whether the disk diffusion test could be improved or whether all susceptibilities of campylobacters should be tested using an MIC-based method.

Ribosomal mutations as the main cause of macrolide resistance in Campylobacter jejuni and Campylobacter coli.

The aim of this study was to examine macrolide resistance mutations in Campylobacter species. In 76 strains studied, point mutation A to G at position 2059 of the 23S rRNA gene was detected in 30 of the 33 erythromycin-resistant strains. An amino acid insertion in the ribosomal protein L22 was found in one resistant strain without a 23S rRNA mutation. The A2059G mutation is the main cause of macrolide resistance in Campylobacter species.

Antimicrobial susceptibilities of multidrug-resistant Campylobacter jejuni and C. coli strains: in vitro activities of 20 antimicrobial agents.

There is a paucity of information regarding antimicrobial agents that are suitable to treat severe infections caused by multidrug-resistant Campylobacter spp. Our aim was to identify agents that are potentially effective against multiresistant Campylobacter strains. The in vitro activities of 20 antimicrobial agents against 238 Campylobacter strains were analyzed by determining MICs by the agar plate dilution method or the Etest. These strains were selected from 1,808 Campylobacter isolates collected from Finnish patients between 2003 and 2005 and screened for macrolide susceptibility by using the disk diffusion test. The 238 strains consisted of 183 strains with erythromycin inhibition zone diameters of < or =23 mm and 55 strains with inhibition zone diameters of >23 mm. Of the 238 Campylobacter strains, 19 were resistant to erythromycin by MIC determinations (MIC > or = 16 microg/ml). Given that the resistant strains were identified among the collection of 1,808 isolates, the frequency of erythromycin resistance was 1.1%. All erythromycin-resistant strains were multidrug resistant, with 18 (94.7%) of them being resistant to ciprofloxacin (MIC > or = 4 microg/ml). The percentages of resistance to tetracycline and amoxicillin-clavulanic acid (co-amoxiclav) were 73.7% and 31.6%, respectively. All macrolide-resistant strains were susceptible to imipenem, meropenem, and tigecycline. Ten (52.6%) multiresistant strains were identified as being Campylobacter jejuni strains, and 9 (47.4%) were identified as being C. coli strains. These data demonstrate that the incidence of macrolide resistance was low but that the macrolide-resistant Campylobacter strains were uniformly multidrug resistant. In addition to the carbapenems, tigecycline was also highly effective against these multidrug-resistant Campylobacter strains in vitro. Its efficacy for the treatment of human campylobacteriosis should be evaluated in clinical trials.

Myopericarditis in a patient with Campylobacter enteritis: a case report and literature review.

Myocarditis sometimes occurs as a complication of bacterial infection, including enteric infections caused by Salmonella, Shigella and Yersinia. Only a few cases of Campylobacter-associated myocarditis are known. We describe a 47-y-old patient with myopericarditis in association with Campylobacter spp. enteritis, and review similar cases previously described in the English literature.

In vitro activities of 11 fluoroquinolones against 226 Campylobacter jejuni strains isolated from Finnish patients, with special reference to ciprofloxacin resistance.

OBJECTIVES: Although Campylobacter jejuni is naturally susceptible to fluoroquinolones, the resistance to these antimicrobials has increased rapidly during the recent years. The aim of this study was to compare the activities of various older and newer fluoroquinolones towards C. jejuni, with special attention on ciprofloxacin-resistant strains. METHODS: We analysed the in vitro activities of 11 fluoroquinolones against 226 C. jejuni strains collected from Finnish patients between 1995 and 2000. RESULTS: Of all 226 C. jejuni strains, 134 (59.3%) were resistant to ciprofloxacin (MIC > or = 4 mg/L), 1 (0.4%) was intermediately resistant (MIC = 2 mg/L) and 91 (40.3%) were ciprofloxacin-susceptible (MIC < or = 1 mg/L). The MIC50 and MIC90 values of ciprofloxacin for the 91 ciprofloxacin-susceptible strains were 0.25 and 0.5 mg/L, respectively. The corresponding MIC50 and MIC90 values of levofloxacin were 0.25 and 0.5 mg/L, and those of moxifloxacin were 0.125 and 0.25 mg/L, these being lower than those of norfloxacin and ofloxacin. The two newer fluoroquinolones, sitafloxacin and clinafloxacin, exhibited the lowest MIC50 and MIC90 values: 0.016 and 0.064 mg/L of sitafloxacin and 0.032 and 0.125 mg/L of clinafloxacin, respectively. Sitafloxacin and clinafloxacin exhibited the lowest MIC50 and MIC90 values also for the 134 ciprofloxacin-resistant C. jejuni strains: 0.25 and 1 mg/L of sitafloxacin and 1 and 4 mg/L of clinafloxacin, respectively. CONCLUSIONS: Of the newer fluoroquinolones presently under development, sitafloxacin is in vitro highly effective towards C. jejuni, with low MIC values also for the ciprofloxacin-resistant strains. Sitafloxacin might be a candidate for clinical trials on campylobacteriosis.

Multidrug resistance in Campylobacter jejuni strains collected from Finnish patients during 1995-2000.

OBJECTIVES: The resistance of Campylobacter jejuni to fluoroquinolones is increasing globally. This study was performed to delineate those antimicrobial agents that are effective in vitro against ciprofloxacin-resistant C. jejuni isolates and potentially suitable for the treatment of severe disease when fluoroquinolone resistance or multidrug resistance is known or suspected. METHODS: During 1995-2000 we collected 376 C. jejuni strains, of which 354 were of foreign origin from multiple countries and 22 were of domestic origin. The MICs of 12 antimicrobial agents against the isolates were determined. RESULTS: Of the 376 strains, 174 (46%) were resistant to ciprofloxacin. Among other antimicrobials, resistance was most common to tetracycline (46%) and ampicillin (17%). Of the ciprofloxacin-resistant strains, 68% and 25%, respectively, were resistant to tetracycline and ampicillin, and 3% were resistant to erythromycin, gentamicin or clindamycin. One (0.6%) ciprofloxacin-resistant isolate was resistant to co-amoxiclav and none was resistant to imipenem. Resistance to three or more antimicrobial groups was detected in 22% of the isolates. Multidrug resistance was significantly associated with ciprofloxacin resistance (33% versus 12%; P<0.01). Eight (2%) strains were resistant to macrolides, of which 75% were also resistant to ciprofloxacin, but none was resistant to co-amoxiclav or imipenem. CONCLUSIONS: Macrolides still appear to be the first-choice alternative for suspected C. jejuni enteritis, if antimicrobial treatment is needed. The in vitro susceptibilities suggest that clinical trials to treat enteritis caused by multidrug-resistant C. jejuni with co-amoxiclav, and life-threatening infections with a carbapenem, may be valuable.