Microbiological Studies on Clostridium Perfringens Type (A)

Background:Clostridium perfringens, a rapid-growing pathogen known to secrete an arsenal of >20 virulent toxins, has been associated with intestinal diseases in both animals and humans throughout the past century. Gas gangrene is synonymous with myonecrosis and is a highly lethal infection of deep soft tissue, caused by Clostridium species, with Clostridium perfringens being the most common. Clostridium perfringens is one of the most common cause’s gas gangrene that is associated with broadspectrum antibiotic treatment. The objectives of the present article were focusing on the diagnosis of Clostridium perfringens from wound swabs from patients suffering from gas gangrene associated with antibiotics treatment and determining the best antibiotics that can inhibit the growth of Clostridium perfringens.Materials and methods: Four hundred isolates were collected from Fever Abbassia hospital in Cairo, Egypt. Clinical samples were collected from swab wounds of limbs. Samples were taken during the period of the first of Oct 2017 till the end of Oct 2018. From four hundred cases collected, Two hundred twenty-eight identified as gas gangrene (57%), One hundred twenty-eightidentified as Clostridium perfringens type(A) (56.1%) confirmed by biochemical tests (Gram stain, Catalase test, Motility test, Skim milk test, Urease test, Indole test, Nagler reaction.) and Multiplex PCR. In this study, we are used different media such as (Thioglycalate broth, Reinforced Clostridial Agar, Blood agar, TryptoseSulfiteCycloserine Agar, Robertson's cooked meat media).  The antibiotic sensitivity against CL.perfringens is assayed by two methods, disc diffusion Method and Determination of (MIC) using E-Test by following the reference standards established by the Clinical and Laboratory Standards Institute ( CLSI)  and British Society for Antimicrobial Chemotherapy  (BSAC). The antimicrobial susceptibility test was performed for all of the isolates (128) by disc diffusion method using antibiotics (Chloramphenicol (30 μg /disc), Cefoxitin (10 μg /disc), Clindamycin (2 μg /disc), Metrondiazole (5 μg /disc), Tetracyclin (30 μg /disc), Rifampin (5 μg /disc), Penicillin (10 μg /disc) and Piperacillin (30 μg /disc) and by E- test (MIC) (0.016- 256µg/1ml) method using antibiotics (Chloramphenicol, Cefoxitin, Clindamycin, Penicillin G, Tetracycline and Piperacillin) on blood agar and incubate anaerobically in Anaerobic jar for 24 hrs. at 37 ˚C. As Rifampin and Metrondiazole breakpoints (E-Test) for Clostridium perfringenhave not yet been established by either EUCAST or CLSI.Results and conclusion: In this study, isolates Clostridium perfringens from selective media TSC Agar or Blood agar and doing biochemical tests, the antibiotics susceptibility test showing that disc diffusion method and MIC method give the same result. Susceptibility tests were done on (128) isolates of CL.perfringens by E-Test to determine MIC.Our results  revealed that the percentage of sensitivity of CL.perfringens to( Chloramphenicol , Cefoxitin, Clindamycin, Rifampin, Penicillin G, Metrondiazole  and Tetracyclin) are (53.9% , 53.1% , 52.3 % , 41.4 % , 35.9 % , 21.1 %  , 14.1 % ) respectively . On the other hand, the data revealed that CL.perfringens was resistant to PRL ( 100%). Future advanced research should participate in the prevention of C. perfringens gas gangrene outbreaks