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PubMed2002

Analysis of macrolide-lincosamide-streptogramin B (MLS(B)) resistance determinant in strains of Clostridium difficile.

The macrolide-lincosamide-streptogramin B (MLSB) resistance determinants have been detected among Clostridia in both C. perfringens and C. difficile strains. Previous studies have shown that MLSB-resistant C. difficile strains can be differentiated by specific hybridizing bands using an erm(B) probe. A recent study has demonstrated that C. difficile 630, a strain highly resistant to clindamycin and erythromycin (MIC > or = 256 ml/L), showing a hybridizing band at 9.7 kb, contains two copies of an erm(B) gene. It was also hypothesized that C. difficile 630 erm(B) determinant has arisen from a progenitor, represented by the C. perfringens CP592 determinant, which contains only one copy of an erm(B) gene that differs from C. difficile 630 erm(B) for seven nucleotide substitutions. To investigate the possibility that C. difficile strains with hybridizing fragments of different molecular size have an erm(B) determinant not identical to the one described in C. difficile 630, we performed a genetic analysis on the erm(B) determinant in 18 C. difficile strains, isolated from different sources. The results showed a heterogeneity in erm(B) determinant: C. difficile strains with hybridizing bands at 7.3 or 3.7 kb contained only one erm(B) copy, whereas strains with a band at 9.7 kb had two copies. The majority of the toxigenic strains examined was characterized by only one erm(B) copy with a sequence identical to the one found in C. difficile 630 and a lower resistance level for erythromycin (MICs ranging from 16 to 24 ml/L). Differently, some strains had an erm(B) gene identical to the one found in C. perfringens CP592. PCR ribotyping and clustering analysis indicate that the examined resistant strains, except one, belong to the same genetic lineage. These results seem to support the hypothesis of the evolution of the C. difficile 630 erm(B) determinant. The functional significance of one or two copies of erm(B) gene in C. difficile strains should be further investigated.

Anti-Bacterial Agents↗

PubMed1997

Can ischemic colitis be differentiated from C difficile colitis in biopsy specimens?

Pseudomembranous colitis is often caused by Clostridium difficile; however, it may also be due to ischemia. To determine if any histologic features could be used to differentiate C difficile from ischemia, 49 biopsies of pseudomembranous colitis (25 from patients with C difficile colitis and 24 from patients with ischemic colitis) were coded, randomized, and evaluated for the presence of numerous variables, including the amount and distribution of mucosal necrosis, lamina propria hyalinization, and atrophic "micro-crypts." Hyalinization of the lamina propria was seen in 19 cases of ischemia but not in C difficile colitis (p < 0.0001). Atrophic-appearing micro-crypts were seen in 18 ischemic cases and 6 C difficile cases (p < 0.0006). Lamina propria hemorrhage, full-thickness mucosal necrosis, and a diffuse microscopic distribution of pseudomembranes were significantly more common in ischemia than C difficile. Endoscopic examination identified pseudomembranes significantly more often with C difficile than ischemia, while the endoscopic appearance of masses or polyps was seen exclusively in cases of ischemia. The presence of a hyalinized lamina propria appeared to be a specific and sensitive marker for ischemia in colon biopsies with pseudomembranes. The presence of atrophic micro-crypts, lamina propria hemorrhage, full-thickness mucosal necrosis, diffuse involvement of all the surface of all biopsies by pseudomembranes, and the endoscopic impression of a localized process, polyp, or mass were also markers of ischemia, while the endoscopic identification of diffuse pseudomembranes favored the diagnosis of C difficile.

Adult↗

PubMed1996

Ascites associated with antibiotic-associated pseudomembranous colitis.

We report the case of an elderly patient who had ascites due to pseudomembranous colitis and associated hypoalbuminemia. Computed tomography showed diffuse colonic wall thickening. An indium-111 scan to localize the site of infection showed abnormal localization of 111In throughout the colon. Despite treatment, the patient died. Autopsy disclosed no other cause for the ascites, except for possible sepsis. To study the cause of ascites in patients with pseudomembranous colitis, we reviewed our institutions' experience with ascites in association with Clostridium difficile colitis, identifying 16 cases over a 1-year period (which included our case). In most of the other cases, the ascites could be attributed primarily to another mechanism, including portal hypertension, congestive heart failure, and sepsis (intra-abdominal and systemic). We also reviewed the literature regarding the association of ascites with C difficile colitis.

Aged↗

PubMed1996

Epidemiology and typing of Clostridium difficile.

Clostridium difficile is now established as the most common nosocomial enteric pathogen causing pseudomembranous colitis, antibiotic-associated colitis and antibiotic-associated diarrhoea. Antibiotic therapy is the most important risk factor in colonization and infection with C. difficile. However, other factors are involved such as age and underlying illness. The introduction of reliable typing and fingerprinting methods has demonstrated hospital acquisition and cross-infection with C. difficile and has been important in improving our understanding of the epidemiology and pathogenicity of C. difficile.

Anti-Bacterial Agents↗

PubMed1993

In-vitro and in-vivo characterisation of resistance to colonisation with Clostridium difficile.

In hamsters, resistance to colonisation by Clostridium difficile appears to be mediated by micro-organisms that are present in the gut in relatively low concentrations. Small amounts of normal caecal contents inhibited the growth of C. difficile when added to cultures in vitro or given to animals which had been treated with clindamycin. Filtrates of caecal contents, frozen and thawed contents and contents diluted to 0.1% wet weight lost their inhibitory properties. However, caecal contents retained their protective capacity after culture for 7 days in vitro. Antibiotic treatment altered resistance to colonisation by only a few species of clostridia. Faeces of animals treated with ampicillin but not clindamycin recovered colonisation resistance after incubation at 37 degrees C in vitro. Since human faeces could also restore colonisation resistance to hamsters, the hamster model may be useful for the study of resistance to colonisation by C. difficile in man.

Animals↗

PubMed1998

Binding of Clostridium difficile toxin A to human milk secretory component.

Toxigenic Clostridium difficile is isolated from a majority of healthy human infants. The exact mechanism of asymptomatic colonisation is unclear; however, previous studies in this laboratory have shown that components of both the immunoglobulin and non-immunoglobulin fractions of human milk bind to toxin A and prevent its interaction with hamster intestinal brush border membranes (BBMs). Secretory IgA (sIgA) is the primary immunoglobulin found in human milk. As sIgA resists digestion in the infant stomach and passes at high levels into the colon, its ability to bind toxin A was the subject of this investigation. Purified sIgA in concentrations at and below those found in human milk inhibited the binding of toxin A to purified BBM receptors. Heating sIgA to 100 degrees C for 5 min did not affect its inhibitory activity. IgM, IgG and serum IgA did not appreciably inhibit the binding of toxin A to BBM receptors. SDS-PAGE separated sIgA into three major bands: secretory component, heavy chains and light chains. Autoradiography with radiolabelled toxin A revealed that toxin A bound to the secretory component (SC) of sIgA. When the three purified subunits of sIgA were coated on to microtitration wells, SC bound significantly more toxin A than the heavy or light chains of sIgA. Purified SC also inhibited toxin binding to receptors in a dose-dependent fashion similar to sIgA. The heavy and light chains of sIgA did not inhibit toxin A receptor binding. Removing carbohydrates from sIgA and SC by enzymic digestion showed that toxin A binds much less to deglycosylated SC than to glycosylated SC. These data suggest that SC in human milk binds to toxin A and may function as a receptor analogue, protecting human infants against C. difficile-associated disease.

Animals↗

PubMed1995

A defined growth medium for Clostridium difficile.

Minimal requirements of amino acids and vitamins were determined in chemically defined medium for five strains of Clostridium difficile. Cysteine, isoleucine, leucine, proline, tryptophan and valine were essential amino acids for growth of C. difficile. Arginine, glycine, histidine, methionine and threonine enhanced growth. Biotin, pantothenate and pyridoxine were essential vitamins. A defined medium containing the minimal requirements of amino acids and vitamins produced a rapid and heavy growth which was comparable to that in modified brain heart infusion, a complex medium. Adenine was able to substitute for glycine and threonine, suggesting that the two amino acids may be utilized as precursors of purine nucleotides. The defined medium developed here will assist physiological and biochemical studies on C. difficile.

Adenine↗

PubMed2001

Medical diagnoses and procedures associated with clostridium difficile colitis.

OBJECTIVES: The aim of this study was to examine the associations of Clostridium difficile colitis with other comorbid conditions and procedural interventions among hospitalized patients. METHODS: The Patient Treatment File of the Department of Veterans Affairs contains the computerized records of all inpatients treated in 172 Veterans Affairs hospitals distributed throughout the United States. The computerized medical records of 15,091 cases with C. difficile colitis and 61,931 controls without the diagnosis were extracted from the annual files between 1993 and 1998. In a multivariable logistic regression, the occurrence of C. difficile colitis served as outcome variable, whereas the occurrences of other diagnoses or procedures served as predictor variables. RESULTS: The total numbers of diagnoses in the case and control group were 136,840 and 465,972, respectively. The numbers of procedures were 75,479 and 129,612, respectively. C. difficile colitis was significantly associated with HIV infection, candidiasis, malignant neoplasm and chemotherapy, malnutrition, pneumonia, aspiration pneumonitis, intestinal obstruction, diverticulitis, renal failure, urinary tract infection, decubitus, and osteomyelitis. Interventional procedures involving the respiratory tract, bone marrow biopsy, arterial and venous catheterization, urinary catheterization, dialysis, gastrostomy tube, and physical therapy were also frequently associated with the development of C. difficile colitis. CONCLUSIONS: These associations reflect the influence of causal relationships (such as the use of antibiotics and chemotherapy), an increased risk of exposure to C. difficile among immobilized bedridden patients with chronic disease states, or a general system failure in patients with end-stage disease. Knowledge of such associations could help to alert physicians to an increased risk of C. difficile colitis among particular groups of susceptible patients.

Aged↗

PubMed1991

In vitro activities of ramoplanin and four glycopeptide antibiotics against clinical isolates of Clostridium difficile.

Seventy strains of Clostridium difficile, all isolated from symptomatic patients, were found to be uniformly susceptible to ramoplanin, a new glycolipodepsipeptide antibiotic, and to four glycopeptides (vancomycin, teicoplanin, and two semisynthetic teicoplanin derivatives). Ramoplanin is recommended for further evaluation in the treatment of C. difficile-associated disease.

Anti-Bacterial Agents↗

PubMed2005

Increased rate of DNA recovery from United Kingdom epidemic Clostridium difficile PCR ribotype 1 strains stored cryogenically.

We noted that some Clostridium difficile isolates are nonrecoverable after frozen storage and so used molecular typing analysis to characterize DNA from these strains. The recovery rate of C. difficile PCR ribotype 1 was statistically significantly greater than that of other strains. This observation has implications for C. difficile epidemiological studies.

Clostridioides difficile↗

PubMed2005