agar and Enterocolitis--Pseudomembranous

ChromID Clostridium difficile agar (IDCd; bioMérieux SA, France) is a recently developed chromogenic medium for rapid and specific isolation of C. difficile. We compared the performance of IDCd with that of Clostridium difficile Selective Agar (CDSA).. A total of 530 fresh stool specimens were collected from patients with clinical signs compatible with C. difficile infection, and cultures for C. difficile were performed on IDCd and CDSA. C. difficile colonies were identified by spore staining, odor, use of an ANI identification test kit (bioMérieux SA), and multiplex PCR for tcdA, tcdB, and tpi.. The concordance rate between IDCd and CDSA was 90.6% (480/530). The positivity rates on IDCd on days 1 and 2 (55.6% and 85.0%, respectively) were significantly higher than those on CDSA (19.4% and 75.6%, respectively) (P<0.001 for day 1 and P=0.02 for day 2), but the detection rates on IDCd and CDSA on day 3 were not different (89.4% vs. 82.8%, P=0.0914). On day 3, the recovery rates for non-C. difficile isolates on IDCd and CDSA were 30.2% (160/530) and 22.1% (117/530), respectively (P=0.0075). Clostridium spp. other than C. difficile were the most prevalent non-C. difficile isolates on both media.. The culture positivity rates on IDCd and CDSA were not different on day 3 but IDCd may allow for rapid and sensitive detection of C. difficile within 2 days of cultivation.

Topics: Agar; Bacterial Proteins; Bacterial Toxins; Clostridioides difficile; DNA, Bacterial; Enterocolitis, Pseudomembranous; Enterotoxins; Feces; Humans; Multiplex Polymerase Chain Reaction; Reagent Kits, Diagnostic; Triose-Phosphate Isomerase

The rapidly changing epidemiology of Clostridium difficile infection highlights the need for improved and continuing surveillance involving stool culturing to enable molecular tracking. Culture of C. difficile can be difficult and time consuming. In this report ChromID C. difficile agar (CDIF) was compared to cycloserine-cefoxitin-fructose-egg-yolk agar which contained 0.1% sodium taurocholate (TCCFA) as a germinant.. All ribotypes of C. difficile tested (n=90) grew well on CDIF within 24 h and most gave characteristic small irregular black colonies with a raised umbonate profile. Counts from standard suspensions of C. difficile at 24 h (p<0.005) and 48 h (p=0.01) were significantly higher on CDIF than on TCCFA. Similar results were achieved after alcohol shock. When temperature shock was used to differentiate vegetative cells and spores, the total number of culturable and vegetative cells on CDIF was significantly higher than on TCCFA (culturable cells, p=0.003 at 24 h and p=0.002 at 48 h; vegetative cells, p=0.0003 at 24 h and p=0.0002 at 48 h).. These data suggest that CDIF is a better medium for the recovery of vegetative C. difficile than TCCFA and equal to TCCFA for spore recovery.

Topics: Agar; Cefoxitin; Clostridioides difficile; Cycloserine; Enterocolitis, Pseudomembranous; Ethanol; Fructose; Humans; Microbiological Techniques; Ribotyping; Temperature; Time Factors

The effects of the inoculum, pH, cation concentrations, and different lots of commercial media on the in vitro susceptibility of Clostridium difficile to fidaxomicin were examined. Of the factors evaluated, only pH alterations influenced the activity of fidaxomicin against C. difficile, noticeably reducing its activity at higher pH (> or =7.9).

Topics: Agar; Aminoglycosides; Anti-Bacterial Agents; Cations; Clostridioides difficile; Colony Count, Microbial; Culture Media; Enterocolitis, Pseudomembranous; Fidaxomicin; Glycosides; Humans; Hydrogen-Ion Concentration; In Vitro Techniques; Microbial Sensitivity Tests

Clostridium difficile plays an essential role in causing pseudomembranous colitis. We looked for the presence of these bacteria in the stools of 169 hospitalized patients and 38 nurses from wards with cases of diarrhea (207 subjects). The study was divided into three parts. In the first part, we compared three methods for isolating Clostridium difficile from stool samples: pre-selection with heat-shock, direct plating on Cycloserine-Cefotaxime-Fructose Agar (CCFA) and culturing in a selective broth medium. Final identification of Clostridium difficile was achieved by gas-chromatography and ApiZym. From the 207 consecutively obtained stool specimens, Clostridium difficile was isolated in 108 (52%) when pre-treated by heat-shock compared to only 26 (13%) when plated on modified CCFA and 23 (11%) when cultured in selective broth medium. Pre-selection significantly increases the isolation rate for Clostridium difficile and should be used in further epidemiological research. In the second part of our study, a retrospective review of subjects' records showed that the heat-shock method detected Clostridium difficile in all age groups at a higher rate than the other methods. In the third part of our study, we typed the 157 isolates of Clostridium difficile strains by protein patterns using SDS-PAGE, and 16 distinct groups were identified. In 19 cases different Clostridium difficile strains were found in the same subject by SDS-PAGE. Finally, the isolated strains were compared with strains from Brussels and Freiburg. Matching patterns were noted in only three cases.

Topics: Adolescent; Adult; Agar; Age Factors; Anti-Bacterial Agents; Cefotaxime; Child; Child, Preschool; Chromatography, Gas; Clostridioides difficile; Culture Media; Cycloserine; Diagnosis, Differential; Electrophoresis, Polyacrylamide Gel; Enterocolitis, Pseudomembranous; Feces; Female; Fructose; Hot Temperature; Humans; Infant; Infant, Newborn; Male; Molecular Weight; Retrospective Studies; Risk Factors; Sensitivity and Specificity; Serotyping

Vibrio alginolyticus was isolated from the "rice water" diarrheal stool of a female patient with acute entero-colitis, and from the trout roe which she ate. Subsequently, it was clearly demonstrated that, besides of Vibrio cholerae and Vibrio parahaemolyticus, Vibrio alginolyticus was also enteropathogenic for humans. Additionally, we described the difference in the colony formation on some commercial thiosulfate citrate bile salts sucrose (TCBS) agars when Vibrio alginolyticus and Vibrio cholerae were tested.

Topics: Agar; Culture Media; Enterocolitis, Pseudomembranous; Female; Humans; Middle Aged; Vibrio