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Updated: Apr 1, 2026
Enrichment and Detection of Clostridium perfringens Toxinotypes in Retail Food Samples
Published on: October 18, 2019
Antibiotic Sensitivity of Clostridium perfringens Isolated From Faeces in Tabriz, Iran
Mohammad Taghi Akhi1, Saeid Bidar Asl2, Tahereh Pirzadeh2
1Research Center of Infectious and Tropical Diseases, Tabriz University of Medical Sciences, Tabriz, IR Iran ; Department of Bacteriology and Virology, School of Medicine, Tabriz University of Medical Sciences, Tabriz IR Iran.
Background:
Clostridium perfringens, a Gram-positive, anaerobic bacterium that produces at least 16 virulence factors including 12 toxins (α-ν), enterotoxin, hemolysin and neuraminidase, can create variable pathogenic condition, ranging from a food poisoning to life-threatening myonecrosis. Among C. perfringens strains, resistance to the drug choices such as penicillin as well as to alternatives of penicillin like metronidazole and clindamycin has also been observed.
Objectives:
The aim of this study was to determine the resistance of isolated toxigenic and non-toxigenic C. perfringens strains against common antimicrobial agents.
Materials And Methods:
In this descriptive study, a total of 136 stool specimens were collected. At first, cooked meat medium enrichment method was performed on samples at 45°C. Thereafter, a loopful of the enriched culture was transferred to blood agar and incubated anaerobically at 37°C for 24-72 hours. Colonies with double zone of hemolysis were identified by different biochemical tests such as phospholipase C (lecithinase) test, indole and urease production. The Minimum Inhibitory Concentration (MIC) for common antibiotics was determined by Etests (Epsilometer) and duplex Polymerase Chain Reaction (PCR) reaction was performed with specific primers for amplification of cpe (426 bp) and plc (283 bp) Genes.
Results:
Of 136 stool samples including diarrhea [48] and non-diarrhea [88] ones, 83 (61.02%) C. perfringens were cultured. Of these 83, 79 C. perfringens isolates showed the alpha-toxin (phospholipase C) production gene by PCR. Respectively, 3 (9.09%) and 2 (4.34%) cpe genes were present in diarrhea and non-diarrhea samples. Of 79 isolates of C. perfringens, 34 (43.03%) cases showed no resistance, 18 (22.78%) had one resistance and 27 (34.17%) isolates had multiple resistance to imipenem, metronidazole, ceftriaxone, clindamycin, chloramphenicol, and penicillin.
Conclusions:
Periodic evaluation of antimicrobial susceptibility for C. perfringens should be performed. Harboring of enterotoxigenic C. perfringens in individuals not necessarily results in diarrhea.
Insights
Clostridium perfringens exhibits concerning antimicrobial resistance, with over half of isolates showing resistance to common antibiotics. Periodic monitoring of C. perfringens susceptibility is crucial for effective treatment strategies.
Area of Science:
- Microbiology
- Infectious Diseases
- Antimicrobial Resistance
Background:
- Clostridium perfringens is a versatile Gram-positive anaerobic bacterium producing numerous toxins, causing conditions from food poisoning to myonecrosis.
- Emerging resistance to penicillin, metronidazole, and clindamycin in C. perfringens necessitates further investigation.
Purpose of the Study:
- To determine the antimicrobial resistance patterns of toxigenic and non-toxigenic Clostridium perfringens strains.
- To assess the prevalence of specific virulence genes, including enterotoxin (cpe) and phospholipase C (plc), in C. perfringens isolates.
Main Methods:
- Cooked meat medium enrichment followed by anaerobic incubation on blood agar for C. perfringens isolation from 136 stool samples.
- Biochemical tests (lecithinase, indole, urease) and duplex Polymerase Chain Reaction (PCR) for identifying C. perfringens and detecting cpe and plc genes.
- Etest (Epsilometer) method to determine the Minimum Inhibitory Concentration (MIC) against various common antibiotics.
Main Results:
- A total of 83 C. perfringens strains were isolated, with 79 possessing the alpha-toxin (plc) gene.
- The enterotoxin gene (cpe) was detected in 3.03% of diarrhea and 4.34% of non-diarrhea samples.
- Of the 79 C. perfringens isolates, 43.03% showed no resistance, 22.78% had one resistance, and 34.17% exhibited multiple drug resistances.
Conclusions:
- Significant antimicrobial resistance, including multiple resistances, was observed in C. perfringens isolates.
- The presence of enterotoxigenic C. perfringens does not always correlate with diarrheal symptoms.
- Regular surveillance of antimicrobial susceptibility in C. perfringens is recommended to guide clinical treatment.
