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PubMed1991

Electron microscopic investigation of lysogeny of Clostridium difficile strains isolated from antibiotic-associated diarrhea cases and from healthy carriers.

An electron microscopic investigation was performed on 28 Clostridium difficile strains isolated from 15 antibiotic-associated diarrhea cases and from 13 healthy infants. Through the use of supernatants of the cultures induced by mitomycin C (1 or 3 micrograms/ml), 18 of the 28 C. difficile strains proved to harbor phage particles with a different morphology and size. Most of the phages revealed belonged in phage group B1 with a non-contractile tail. Seven of the 11 toxin-producing, lysogenic C. difficile strains carried a defective phage structure (120 nm-long tail with an incomplete head capsule) alone or together with other normal phages. With different C. difficile strains as indicator, plaque formation could not be detected in any of the lysates.

Anti-Bacterial Agents↗

PubMed2003

Nondigestible oligosaccharides enhance bacterial colonization resistance against Clostridium difficile in vitro.

Clostridium difficile is the principal etiologic agent of pseudomembranous colitis and is a major cause of nosocomial antibiotic-associated diarrhea. A limited degree of success in controlling C. difficile infection has been achieved by using probiotics; however, prebiotics can also be used to change bacterial community structure and metabolism in the large gut, although the effects of these carbohydrates on suppression of clostridial pathogens have not been well characterized. The aims of this study were to investigate the bifidogenicity of three nondigestible oligosaccharide (NDO) preparations in normal and antibiotic-treated fecal microbiotas in vitro and their abilities to increase barrier resistance against colonization by C. difficile by using cultural and molecular techniques. Fecal cultures from three healthy volunteers were challenged with a toxigenic strain of C. difficile, and molecular probes were used to monitor growth of the pathogen, together with growth of bifidobacterial and bacteroides populations, over a time course. Evidence of colonization resistance was assessed by determining viable bacterial counts, short-chain fatty acid formation, and cytotoxic activity. Chemostat studies were then performed to determine whether there was a direct correlation between bifidobacteria and C. difficile suppression. NDO were shown to stimulate bifidobacterial growth, and there were concomitant reductions in C. difficile populations. However, in the presence of clindamycin, activity against bifidobacteria was augmented in the presence of NDO, resulting in a further loss of colonization resistance. In the absence of clindamycin, NDO enhanced colonization resistance against C. difficile, although this could not be attributed to bifidobacterium-induced inhibitory phenomena.

Anti-Bacterial Agents↗

PubMed1991

Cytokine response by human monocytes to Clostridium difficile toxin A and toxin B.

Clostridium difficile toxins A and B isolated from strain VPI 10463 were tested for induction of cytokine release by human monocytes. Toxin B at 10(-12) M activated human monocytes as measured by release of interleukin-1 (IL-1), tumor necrosis factor (TNF), or IL-6. These effects of toxin B were heat labile (51 degrees C, 30 min). Toxin B was as effective as bacterial lipopolysaccharides in inducing IL-1 beta but less effective in inducing TNF or IL-6. Toxin B and lipopolysaccharides were synergistic in induction of IL-1 beta, TNF, and IL-6. The toxin A preparation used was 1,000-fold less active than toxin B. Apart from the difference in activity, the two toxins showed identical patterns of reaction and there was no synergism between them. A short pulse with toxin B was sufficient to trigger IL-1 release. Toxin B was also extremely toxic for monocytes. The toxicity and the induced proinflammatory monokines (IL-1 and TNF) may contribute to the pathogenic mechanisms of C. difficile infection and pseudomembranous colitis.

Bacterial Proteins↗

PubMed2000

Clostridium difficile recombinant toxin A repeating units as a carrier protein for conjugate vaccines: studies of pneumococcal type 14, Escherichia coli K1, and Shigella flexneri type 2a polysaccharides in mice.

Unlike the native protein, a nontoxic peptide (repeating unit of the native toxin designated rARU) from Clostridium difficile toxin A (CDTA) afforded an antigen that could be bound covalently to the surface polysaccharides of pneumococcus type 14, Shigella flexneri type 2a, and Escherichia coli K1. The yields of these polysaccharide-protein conjugates were significantly increased by prior treatment of rARU with succinic anhydride. Conjugates, prepared with rARU or succinylated (rARUsucc), were administered to mice by a clinically relevant dosage and immunization scheme. All conjugates elicited high levels of serum immunoglobulin G both to the polysaccharides and to CDTA. Conjugate-induced anti-CDTA had neutralizing activity in vitro and protected mice challenged with CDTA, similar to the rARU alone. Conjugates prepared with succinylated rARU, therefore, have potential for serving both as effective carrier proteins for polysaccharides and for preventing enteric disease caused by C. difficile.

Animals↗

PubMed2006

Typing and subtyping of Clostridium difficile isolates by using multiple-locus variable-number tandem-repeat analysis.

Using the genomic sequence of Clostridium difficile strain 630, we developed multiple-locus variable-number tandem-repeat analysis (MLVA) with automated fragment analysis and multicolored capillary electrophoresis as a typing method for C. difficile. All reference strains, representing 31 serogroups, 25 toxinotypes, and 7 known subtypes of PCR ribotype 001, could be discriminated from each other. Application of MLVA to 28 isolates from 7 outbreaks due to the emerging hypervirulent PCR ribotype 027-pulsed-field gel electrophoresis type NAP1 resulted in recognition of 13 clusters. Additionally, 29 toxin A-negative, toxin B-positive isolates belonging to PCR ribotype 017 from eight different countries revealed eight country-specific clusters. MLVA is a highly discriminatory genotyping method and a new tool for subtyping of newly emerging variants of C. difficile.

Automation↗

PubMed2006

Impact of a piperacillin-tazobactam shortage on antimicrobial prescribing and the rate of vancomycin-resistant enterococci and Clostridium difficile infections.

UNLABELLED: States in 2002 on antimicrobial prescribing and associated rates of vancomycin-resistant enterococci (VRE) and Clostridium difficile infections.Design. Retrospective chart review.Setting. University-affiliated medical center. Measurements and Main Results. Microbiologic reports, patient demographics, and antimicrobial utilization were evaluated for patients admitted 6 months before the shortage (March 1-August 31, 2001) and for 6 months during the shortage (March 1-August 31, 2002). Significant increases in usage of alternative mu-lactamase inhibitor combinations, cefepime, levofloxacin, vancomycin, clindamycin, and metronidazole were observed during the shortage; in contrast, a significant decrease in the use of ceftriaxone took place. No change in the rate of VRE infection was observed from before to during the piperacillin-tazobactam shortage. However, a paradoxical 47% decrease in the rate of C. difficile colitis was documented during the shortage. Subsequent multivariate analyses suggested the reduced use of ceftriaxone and increased use of levofloxacin, but not the reduced use of piperacillin-tazobactam, correlated with the decreased rate of C. difficile infections. CONCLUSION: The piperacillin-tazobactam shortage was associated with significant changes in antimicrobial prescribing, which resulted in a significant reduction in the rate of C. difficile but not VRE infections.

Anti-Bacterial Agents↗

PubMed2001

Suspected Clostridium difficile-associated diarrhea in two cats.

Two adult cats from the same household developed acute diarrhea. Clostridium difficile toxins were detected in the feces of both cats, whereas other recognized causes of diarrhea were not identified. Supportive medical treatment and metronidazole were administered and both cats responded well. A fecal sample obtained from 1 of the affected cats after treatment and a fecal sample obtained from a clinically normal cat in the household did not contain C difficile toxins. The role of C difficile in enteric disease in cats has not been extensively studied and is unclear; however, our findings suggest that toxigenic strains of C difficile may cause diarrhea in cats.

Animals↗

PubMed2001

A prospective study of the roles of clostridium difficile and enterotoxigenic Clostridium perfringens in equine diarrhoea.

Faecal samples from adult horses and from foals with diarrhoea or with normal faeces were evaluated for the presence of Clostridium difficile, C. difficile toxins, C. perfringens enterotoxin (CPE) and C. perfringens spore counts. Clostridium difficile was isolated from 7/55 horses (12.7%) and 11/31 foals (35.5%) with colitis, but from 1/255 normal adults (0.4%) and 0/47 normal foals (P<0.001). Clostridium difficile toxins A and/or B were detected in 12/55 diarrhoeic adults (21.8%) and 5/30 diarrhoeic foals (16.7%) but in only 1/83 adults (1.2%) and 0/21 foals with normal faeces (P<0.001 and P<0.05, respectively). Clostridium perfringens enterotoxin was detected in 9/47 diarrhoeic adults (19%) and 8/28 diarrhoeic foals (28.6%), but was not detected in 47 adult horses (P<0.002) or 4 foals (P = 0.22) with normal faeces. The positive predictive value of isolation of C. perfringens with respect to the presence of CPE was only 60% in adult horses and 64% in foals. There was no association between total C. perfringens spore count and CPE in the faeces. The overall mortality rate from colitis was 22% for adult horses and 18% for foals. Clostridium difficile toxin-positive adult horses with colitis were less likely to survive than C. difficile-negative horses with colitis (P = 0.03). This study provides further evidence that C. difficile and enterotoxigenic C. perfringens are associated with equine enterocolitis.

Animals↗

PubMed1992

Clostridium difficile infection--a poor prognostic sign in uremic patients?

Uremia has been reported as a risk factor for the occurrence of infection with Clostridium difficile. During the two-year period 1984-86, 110 episodes of Clostridium difficile infection were encountered in 70 patients on a nephrology ward. Sixty-two patients had chronic renal failure and eight had acute renal failure. Sixty-seven of the patients were uremic and were treated with hemodialysis (n = 35), CAPD (n = 21), intermittent peritoneal dialysis (n = 6) or conservatively with a low protein diet (n = 5). Most of the patients were female (n = 41) and elderly (64 +/- 2 years). Malnutrition was common as indicated by low serum albumin concentrations (26 +/- 1 g/l) prior to the Clostridium difficile infection. Clostridium difficile infection was confirmed by stool culture and/or cytotoxin assay. Asymptomatic infections were found in eight patients. The highest relative risks of subsequent Clostridium difficile infection were calculated for patients treated with cephalosporins and isoxazolyl penicillins. All patients were treated with vancomycin, which often resulted in a dramatic improvement. One to six relapses of Clostridium difficile infection were observed in 22 of the patients. Sixty of the original 70 patients died during the five-year follow-up period. Thirty-four patients died during the first year of follow-up. Seven patients were transplanted, two are still on CAPD treatment and one has only moderate chronic renal failure (serum creatinine 200 mumol/l). Elderly debilitated uremic patients are especially susceptible to infection with Clostridium difficile which may be a poor prognostic sign in chronic renal failure.(ABSTRACT TRUNCATED AT 250 WORDS)

Clostridioides difficile↗

PubMed1996

The lack of value of repeated Clostridium difficile cytotoxicity assays.

OBJECTIVE: To determine the value of repeated Clostridium difficile cytotoxicity assays (CA). DESIGN: All CAs performed during 1993 were retrospectively reviewed and correlated with clinical data. Assays were grouped into episodes, which were defined as one or more successive tests performed on a single patient within 7 days or less of each other. SETTING: A 751-bed tertiary care facility. PATIENTS: All patients with Clostridium difficile CAs submitted to the microbiology laboratory. RESULTS: There were 947 episodes with two or more CAs. In 15 of these episodes, a negative CA result was followed by a positive result, and in 25 cases, a positive result was followed by a negative one. We reviewed the clinical data for these cases. Of the 947 episodes with two or more CAs, the repeated assays provided new information that was used in patient care in fewer than nine cases. Repeated testing within 7 days of an initial CA accounted for 36% of all assays performed, but provided clinically useful information in only about 1% of cases. CONCLUSIONS: Clostridium difficile CAs should not be repeated within a 7-day period.

Bacterial Toxins↗

PubMed1992

Epidemiological aspects of infections caused by Bacteroides fragilis and Clostridium difficile.

Bacteroides fragilis and Clostridium difficile are two of the most common anaerobes associated with human disease. Studies on the epidemiology of Bacteroides fragilis are limited and are based predominantly on serogrouping, which suggests intraspecies differences. Further studies using newer techniques for typing are required to elucidate the epidemiological characteristics of this important pathogen. By contrast, numerous phenotypic, immunological and molecular methods have been developed for typing and fingerprinting of Clostridium difficile and applied in epidemiological studies to show conclusively that Clostridium difficile is nosocomially acquired and that there is transmission and cross-infection between hospital patients.

Animals↗

PubMed2003

Prolonged ileus as a sole manifestation of pseudomembranous enterocolitis.

BACKGROUND: Pseudomembranous colitis usually manifests as fever and diarrhea in hospitalized patients treated with systemic antibiotics. We present a case that represents a unique variant. CASE PRESENTATION: The 44-year-old man suffered of several weeks of abdominal pain, low-grade fever, nausea, vomiting, and lack of bowel movements. Upper gastrointestinal barium swallow and passage series revealed evidence of severe intestinal hypomotility. A thorough evaluation for the cause of the patient's ileus and abdominal pain was unrevealing, and symptomatic treatment was ineffective. Following the administration of opiates and dietary fiber supplementation the patient's abdominal pain and distention rapidly worsened, requiring an urgent subtotal colectomy. The macroscopic and microscopic appearance of the excised colon as well as results of the colonic cytotoxin essay and fecal enzyme-linked immunosorbent assay essay confirmed the diagnosis of severe Clostridium difficile induced pseudomembranous colitis as the cause of the patient's illness. CONCLUSION: To our knowledge, this is the first reported case of Clostridium-difficile induced disease consisting of prolonged ileus in the absence of diarrhea in a patient not previously taking antibiotics.

Adult↗

PubMedDate not supplied

Clostridium difficile: an update.

Clostridium difficile causes a spectrum of diarrheal illness with the potential for major medical consequences. Although most cases respond quickly to treatment, C. difficile colitis may be severe and life threatening. Recurrent disease represents a particularly challenging problem.

Anion Exchange Resins↗

PubMed2006

Epidemiologic surveillance of Clostridium difficile diarrhea in a freestanding pediatric hospital and a pediatric hospital at a university medical center.

To describe the epidemiology of Clostridium difficile in children, we cultured stool specimens from patients at the Children's Hospital Central California, Madera, CA (CHCC, n = 676) and at the University of California Davis Medical Center Pediatric Hospital, Sacramento, CA (UCDMC-PH, n = 301) for C. difficile, and toxins A and B genes and strain identity of the isolates were determined by polymerase chain reaction assays. A higher percentage of patients from UCDMC-PH were culture positive (148/301, 49%) and colonized with toxigenic strains (45/301, 15%) compared with CHCC (colonized = 178/676, 26%; toxigenic = 96/676, 14%, P < or = .001). Multiple logistic regression analysis showed decreased colonization with inpatient status (odds ratio [OR] = 0.64; 95% confidence interval [CI] = 0.46, 0.89; P = .007) and use of H-2 antagonists (OR = 0.55; 95% CI = 0.36, 0.84; P = .006), whereas underlying conditions (colonization: OR = 1.42; 95% CI = 1.02, 1.96; P = .04; toxin positive: OR = 1.60; 95% CI = 1.04, 2.44; P = .03) and exposure to > or =2 antiinfectives (colonization: OR = 1.56; 95% CI = 1.10, 2.20; P = .01; toxin positive: OR = 1.71; 95% CI = 1.10, 2.66; P = .02) increased colonization. Most isolates appear to be community acquired, although molecular analysis suggests some nosocomial transmission at UCDMC-PH. These data suggest that the epidemiology of colonization with C. difficile in children is different than previously reported.

Carrier State↗

PubMed1997

Acceptor hydroxyl group mapping for calf thymus alpha-(1-->3)-galactosyltransferase and enzymatic synthesis of alpha-D-Galp-(1-->3)-beta-D-Galp-(1-->4)-beta D-GlcpNAc analogs.

The epitope of the acceptor substrate for alpha-(1-->3)-galactosyltransferase from calf thymus has been mapped by using a series of mono-deoxygenated and mono-O-alkylated Type II (beta-D-Ga1p-(1-->4)-beta-D-G1cpNAc) disaccharides. The 4-OH group of the beta-D-galactopyranosyl residue is a key polar group essential for glycosyl transfer, tolerating neither deoxygenation nor O-alkylation. Substitution at positions 6 and 6' by a variety of polar alkyl substituents was readily tolerated, allowing the preparative enzymatic synthesis of a series of trisaccharide derivatives carrying polar substituents on each of these hydroxyl groups. These new analogs are potential inhibitors of Clostridium difficile toxin A and of a human anti-alpha-Gal antibody.

Animals↗

PubMed1998

Silver sulfadiazine induced Clostridium difficile toxic megacolon in a burn patient: case report.

A 53 yr old diabetic male presented with a 34% total body surface area (TBSA) deep partial- and full-thickness burns. On post burn days 4 and 9, all of his burns were excised and grafted. Although he had only been treated with topical antibiotics, he developed Clostridium difficile colitis after his second surgery that progressed to Toxic Megacolon and perforation. The incidence and treatment of Toxic Megacolon secondary to C. difficile is reviewed. Early diagnosis and treatment with colonoscopic decompression may obviate the need for surgery.

Anti-Infective Agents, Local↗

PubMed1994

[Incidence and importance of Clostridium difficile in patients treated with antibacterial therapy or perioperative preventive cefoxitin administration].

Clostridium difficile and/or toxin B were isolated from the faeces of 3 (1.6%) of 184 patients within the first two days of hospitalisation in the University of Tübingen, Gynaecological Clinic. With following stool samples from a total of 117 patients the isolation rate of C. difficile was examined in relation to the treatment with antibiotics. From the first group of 61 patients, who had had gynaecological operations and were not treated with any antibiotic, the isolation rate of C. difficile rose statistically but not significantly from 0% at the time of admission up to 3.3% during hospitalisation. Amongst the second group of 17 patients, who had had gynaecological operations and were treated with various antibiotics for at least three days, the isolation rate was also found to rise insignificantly from 0 to 11.8% (p < 0.1). The third group of 33 patients, who had had gynaecological operations and perioperative prophylaxis with cefoxitin (one to three doses 2 g each), the isolation rate rose significantly from 0 to 12.1% (p < 0.05). The result of this study shows that the bacterial flora of the gut was probably altered even after a very brief application of cefoxitin. In two environmental examinations on the wards of the patients of this study, C. difficile was not isolated in any case.

Adult↗

PubMed1996

Protein-losing enteropathy is associated with Clostridium difficile diarrhea but not with asymptomatic colonization: a prospective, case-control study.

A prospective, case-control study was performed in which enteric protein loss and nutritional status were measured in patients with symptomatic and asymptomatic infections due to Clostridium difficile. Enteric protein loss, measured by elevated levels of fecal alpha1-antitrypsin, was detected in 14 of 20 cases and controls with diarrhea (9 of 10 cases with C. difficile-associated diarrhea and 5 of 10 age-matched control with diarrhea not associated with C. difficile) compared with none of 20 asymptomatic cases and controls (10 colonized cases and 10 noncolonized controls without diarrhea who were matched by age and clinical diagnosis) (P < .0001). Cases and controls with diarrhea had higher prognostic nutritional index values (P = 0.005) and lower levels of serum albumin, transferrin, and cholesterol than did the asymptomatic cases and controls. Decreased nutritional status, measured by increased prognostic nutritional index values, was associated with the presence of diarrhea but not with the presence of C. difficile. Protein-losing enteropathy was associated with C. difficile only in the presence of diarrhea, and we did not detect an increased risk of protein-losing enteropathy or malnutrition as a consequence of asymptomatic colonization with C difficile.

Aged↗