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Distribution of Clostridium Difficile Ribotypes in Macedonian Patients and their Antimicrobial Susceptibility
Kiril Mihajlov1, Aneta Andreska1, Nadica Ristovska1
1Institute of Microbiology and Parasitology, Medical Faculty, Ss Cyril and Methodius University of Skopje, Skopje, Republic of Macedonia.
Background:
Clostridium difficile is a major nosocomial pathogen. In Europe, this bacterium is mostly characterised by PCR ribotyping. Most of the Clostridium difficile infections (CDI) are treated with vancomycin or metronidazole, although prolonged antibiotic use is considered as one of the main risk factors for CDI.
Aim:
This study aimed to detect the presence of various C. difficile ribotypes in hospitalised patients and to investigate their toxigenicity and antibiotic susceptibility.
Material And Methods:
All stool samples obtained from each patient were inoculated on Columbia blood agar and cycloserine cefoxitine fructose agar (CCFA) for isolation of C. difficile. Glutamate dehydrogenase and toxins A and B were investigated by immunochromatographic tests. Final confirmation of the isolates was performed by Vitek 2 and MALDI-TOF. A total of 21 isolates were collected for further investigation. PCR ribotyping was performed as described by Janezic and Rupnik. PCR ribotype profiles were analysed using software (Bionumerics, Applied Maths). Antibiotic susceptibility was determined by E-tests for metronidazole, vancomycin, tetracycline, clindamycin, erythromycin, imipenem, ciprofloxacin and moxifloxacin.
Results:
About 48% of C. difficile isolates belonged to ribotype 001/072. So, this ribotype was the most common ribotype in this study. The remaining 52% of C. difficile isolates consisted of 10 different ribotypes: 017, SLO 160, SLO 187, SLO 120, 255/258, 014/020, 046, 002, 070 and 027. Furthermore, 20 (95.2 %) out of 21 isolates of C. difficile were toxigenic. Toxins A and B were detected simultaneously in 90.5 % of C. difficile isolates. Two isolates from the ribotype 017 were toxin B positive only. Treatments with any of the following antimicrobials: clindamycin, erythromycin, ciprofloxacin and moxifloxacin (as well as many other antibiotics), could be a risk factor for CDI due to the high resistance of the strains in this study. About 90% of the strains from the most common ribotype 001/072 have MICs for clindamycin and erythromycin > 256 µg/ml.
Conclusion:
All strains isolated are highly resistant to ciprofloxacin. All strains were susceptible to vancomycin (median MIC was 0.63 µg/ml) and metronidazole (median MIC was 0.084 µg/ml), so these two antimicrobials remain optimal treatment option for CDI.
Insights
Clostridium difficile infections (CDI) are often caused by resistant strains, but vancomycin and metronidazole remain effective treatments. This study identified common ribotypes and their antibiotic resistance patterns in hospital patients.
Area of Science:
- Medical Microbiology
- Infectious Diseases
- Antimicrobial Resistance
Background:
- Clostridium difficile is a significant cause of hospital-acquired infections.
- PCR ribotyping is a key method for characterizing C. difficile strains in Europe.
- Antibiotic treatment, particularly prolonged use, is a major risk factor for C. difficile infections (CDI).
Purpose of the Study:
- To identify prevalent C. difficile ribotypes in hospitalized patients.
- To assess the toxigenicity of identified C. difficile strains.
- To determine the antibiotic susceptibility profiles of C. difficile isolates.
Main Methods:
- Stool samples were cultured on selective media (Columbia blood agar, CCFA).
- Glutamate dehydrogenase, toxin A, and toxin B were detected using immunochromatographic tests.
- Isolates were confirmed using Vitek 2 and MALDI-TOF, with PCR ribotyping for strain characterization and E-tests for antibiotic susceptibility.
Main Results:
- Ribotype 001/072 was the most common (48%), followed by 10 other ribotypes.
- A high proportion (95.2%) of isolates were toxigenic, with simultaneous detection of toxins A and B in 90.5%.
- Significant resistance was observed against clindamycin and erythromycin (> 256 µg/ml) in ribotype 001/072 strains; all strains were resistant to ciprofloxacin.
Conclusions:
- Vancomycin and metronidazole remain the optimal treatment choices for CDI due to universal susceptibility.
- High resistance to certain antibiotics like ciprofloxacin, clindamycin, and erythromycin highlights the need for careful treatment selection.
- Understanding the prevalence of specific C. difficile ribotypes and their resistance patterns is crucial for infection control.
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