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PubMed1995

Evaluation of formalin-inactivated Clostridium difficile vaccines administered by parenteral and mucosal routes of immunization in hamsters.

Clostridium difficile produces toxins that cause inflammation, necrosis, and fluid in the intestine and is the most important cause of nosocomial antibiotic-associated diarrhea and colitis. We evaluated C. difficile antigens as vaccines to protect against systemic and intestinal disease in a hamster model of clindamycin colitis. Formalin-inactivated culture filtrates from a highly toxigenic strain were administered by mucosal routes (intranasal, intragastric, and rectal) with cholera toxin as a mucosal adjuvant. A preparation of culture filtrate and killed whole cells was also tested rectally. The toxoid was also tested parenterally (subcutaneously and intraperitoneally) and by a combination of three intranasal immunizations followed by a combined intranasal-intraperitoneal boost. Serum antibodies against toxins A and B and whole-cell antigen were measured by enzyme-linked immunosorbent assay, neutralization of cytotoxic activity, and bacterial agglutination. The two rectal immunization regimens induced low antibody responses and protected only 20% of hamsters against death and 0% against diarrhea. The intragastric regimen induced high antibody responses but low protection, 40% against death and 0% against diarrhea. Hamsters immunized by the intranasal, intraperitoneal, and subcutaneous routes were 100% protected against death and partially protected (40, 40, and 20%, respectively) against diarrhea. Among the latter groups, intraperitoneally immunized animals had the highest serum anticytotoxic activity and the highest agglutinating antibody responses. Hamsters immunized intranasally and revaccinated intraperitoneally were 100% protected against both death and diarrhea. Protection against death and diarrhea correlated with antibody responses to all antigens tested. The results indicate that optimal protection against C. difficile disease can be achieved with combined parenteral and mucosal immunization.

Administration, Intranasal↗
PubMed2003

Clostridium difficile vaccine and serum immunoglobulin G antibody response to toxin A.

There is a strong association between serum antibody responses to toxin A and protection against Clostridium difficile diarrhea. A parenteral C. difficile toxoid vaccine induced very-high-level responses to anti-toxin A immunoglobulin G (IgG) in the sera of healthy volunteers. After vaccination, the concentrations of anti-toxin A IgG in the sera of all 30 recipients exceeded the concentrations that were associated with protection in previous clinical studies. Furthermore, the median concentration of serum anti-toxin A IgG in the test group was 50-fold higher than the previous threshold. These findings support the feasibility of using a vaccine to protect high-risk individuals against C. difficile-associated diarrhea and colitis.

Adult↗
PubMed2003

Gastrointestinal complications in the neutropenic patient: characterization and differentiation with abdominal CT.

PURPOSE: To characterize the computed tomographic (CT) findings of gastrointestinal complications in neutropenic patients and to identify CT features that can help differentiate these complications. MATERIALS AND METHODS: Abdominal CT scans obtained during a 6-year period were reviewed retrospectively to identify 76 neutropenic patients with radiologic bowel abnormalities. Scans were analyzed for wall thickening, pneumatosis, wall nodularity, mucosal enhancement, bowel dilatation, ascites, and mesenteric stranding. The location and extent of abnormalities were noted. Independent chart and pathology report reviews were used to determine the patients' final diagnoses: neutropenic enterocolitis (n = 53), Clostridium difficile colitis (n = 14), graft-versus-host disease (n = 7), cytomegaloviral colitis (n = 1), and ischemic bowel (n = 1). Results were assessed with the Student t test for quantitative wall thickness and the chi2 test for the number of patients with each diagnosis who demonstrated each CT finding. RESULTS: Mean bowel wall thickening was greatest in C difficile colitis (12 mm) and least in graft-versus-host disease (5 mm). Pneumatosis was limited to neutropenic enterocolitis (21% [11 of 53 patients]) and bowel ischemia. Wall nodularity was significantly more common (P <.01) in C difficile colitis (36% [five of 14 patients]). In graft-versus-host disease, the rates of mucosal enhancement and bowel dilatation were highest (P <.05) (71% [five of seven patients] and 86% [six of seven patients], respectively). In C difficile colitis, the rates of ascites and mesenteric stranding were highest (57% [eight of 14 patients] and 71% [10 of 14 patients], respectively). Although findings in neutropenic enterocolitis and graft-versus-host disease could involve any bowel segment, C difficile colitis was always limited to the colon. CONCLUSION: Several CT findings can help differentiate specific gastrointestinal complications in neutropenic patients.

Adolescent↗
PubMed1995

Clostridium difficile toxin B is more potent than toxin A in damaging human colonic epithelium in vitro.

Toxin A but not toxin B, appears to mediate intestinal damage in animal models of Clostridium difficile enteritis. The purpose of this study was to investigate the electrophysiologic and morphologic effects of purified C. difficile toxins A and B on human colonic mucosa in Ussing chambers. Luminal exposure of tissues to 16-65 nM of toxin A and 0.2-29 nM of toxin B for 5 h caused dose-dependent epithelial damage. Potential difference, short-circuit current and resistance decreased by 76, 58, and 46%, respectively, with 32 nM of toxin A and by 76, 55, and 47%, respectively, with 3 nM of toxin B, when compared with baseline (P < 0.05). 3 nM of toxin A did not cause electrophysiologic changes. Permeability to [3H]mannitol increased 16-fold after exposure to 32 nM of toxin A and to 3 nM of toxin B when compared with controls (P < 0.05). Light and scanning electron microscopy after exposure to either toxin revealed patchy damage and exfoliation of superficial epithelial cells, while crypt epithelium remained intact. Fluorescent microscopy of phalloidin-stained sections showed that both toxins caused disruption and condensation of cellular F-actin. Our results demonstrate that the human colon is approximately 10 times more sensitive to the damaging effects of toxin B than toxin A, suggesting that toxin B may be more important than toxin A in the pathogenesis of C. difficile colitis in man.

Actins↗
PubMed1996

Community-acquired toxigenic Clostridium difficile diarrhoea in the normoxaemic elderly who have received no antimicrobials: soft evidence for ischaemic colitis?

We report three examples of community-acquired toxigenic Clostridium difficile diarrhoea in elderly patients who had neither received antimicrobial therapy nor been institutionalised. These cases stimulated interest in the non-antimicrobial changes which might predispose the host to C. difficile-related disease and raised the spectre of bowel ischaemia as a possible aetiological factor.

Aged↗
PubMed2002

A survey of agents associated with neonatal diarrhea in Iowa swine including Clostridium difficile and porcine reproductive and respiratory syndrome virus.

This survey was undertaken to determine the relative frequency of agents that are currently associated with neonatal diarrhea in swine, including Clostridium difficile and porcine reproductive and respiratory syndrome virus (PRRSV). The subjects for this study were the first 100 live 1-7-day-old piglets submitted to the Iowa State University Veterinary Diagnostic Laboratory with a clinical signalment of diarrhea, beginning on January 1, 2000. The evaluation of each pig included bacterial culture of a section of ileum, 2 sections of jejunum, and a single section of colon; a fluorescent antibody test (FAT) or immunohistochemistry (IHC) for transmissible gastroenteritis virus (TGEV); ELISA's for rotavirus and C. difficile toxins; IHC for PRRSV; and microscopic examination of ileum, midjejunum, spiral colon, liver, spleen, and lung. Survey results demonstrate a decline in the relative number of diagnoses of TGEV, Escherichia coli, and Clostridium perfringens type C compared with retrospective data. The combined case frequency rate for these 3 pathogens dropped from 70% in 1988 to 21% in 2000. This survey also demonstrated the emergence of C. difficile as an important pathogen of neonatal swine. Clostridium difficle toxin was detected in the colon contents of 29% of the piglets, and at least 1 toxin-positive animal was identified in 55% of the cases. All 29 C. difficile toxin-positive piglets had mesocolonic edema, and colitis was observed in 21 of 29 toxin-positive animals. PRRSV-positive macrophages were detected in the lamina propria of intestinal villi by IHC in 10 piglets with diarrhea. In 6 of these cases, PRRSV was the only pathogen detected. Gross and microscopic lung lesions were not a reliable indicator of PRRSV infection in these neonatal pigs with diarrhea. The addition of tests for C. difficile and PRRSV to a routine neonatal diarrhea diagnostic protocol resulted in a significant increase in thediagnostic success rate on both individual animal and case bases.

Animals↗
PubMed1994

Fecal shedding of Clostridium difficile in dogs: a period prevalence survey in a veterinary medical teaching hospital.

The goal of this study was to determine the fecal prevalence of Clostridium difficile in dogs who were patients at a veterinary medical teaching hospital. Stool specimens collected from 152 dogs (in- and outpatients) were analyzed for the presence of C. difficile. An additional 42 stool specimens were collected and examined from dogs recently housed at local animal shelters. Following culture on selective medium, C. difficile was identified by a latex agglutination test, and the presence of the toxin A and B genes was determined individually by polymerase chain reaction. Clostridium difficile was isolated from the feces of 28 of the veterinary hospital patients (18.4%); isolates from 14 of these patients (50.0%) were toxigenic. Diarrhea was a clinical finding in 5 (35.7%) of the dogs carrying toxigenic isolates of C. difficile, whereas diarrhea was noted in only 2 of 14 dogs (14.3%) shedding nontoxigenic isolates. Three of 14 dogs (21.4%) shedding toxigenic isolates of C. difficile were receiving antibiotics at the time of stool collection, whereas 5 of 14 dogs (37.5%) shedding nontoxigenic strains of C. difficile were receiving antibiotics. The carriage rate of C. difficile was significantly higher for animals categorized as inpatients of the veterinary hospital. The carriage rate also provided evidence for an increased risk for fecal shedding with increasing age. Clostridium difficile was not isolated from any of the 42 dogs recently housed at local animal shelters. This study confirms the presence of toxigenic C. difficile in dogs at a veterinary teaching hospital.(ABSTRACT TRUNCATED AT 250 WORDS)

Age Factors↗
PubMed2000

[Recurrent Clostridium difficile enterocolitis].

Pseudomembranous enterocolitis generally occurs after antibiotic treatment. The standard treatment is oral metronidazol or vancomycin. Nevertheless, relapses of Clostridium difficile enterocolitis are observed in 10-25% of cases. Factors associated with recurrences include endogenous reinfection by spore formation, selective IgG1 or IgA deficiency or infection with mutated strains of Clostridium difficile. Recurrent Clostridium difficile enterocolitis may be treated with repeat oral vancomycin combined with Sacchoromyces boulardii, with intravenous immunoglobulin for severe colitis.

Aged↗
PubMed2003

[Comparison of AP-PCR methods, ribotyping (PCR-ribotyping) and pulsed-field electrophoresis (PFGE) for strains of Clostridium difficile producing toxin B and not producing toxin A].

In this study were used AP-PCR, PCR-ribotyping and pulsed-field elecrophoresis (PFGE) for comparative study of toxin A-negative/toxin B-posi-tive Clostridium difficile strains with deletion in toxin A gen. We investigated nine unrelated clinical strains, isolated from different units and different time from patients suffering to antibiotic associated diarrhea (AAD). We found that toxin A-negative/toxin B-positive C. difficile strains isolated in Poland belonging to a single genotype A, are being similar to the Japanese strains.

Bacterial Toxins↗
PubMed1994

[Nosocomial epidemic outbreak of diarrhea from Clostridium difficile. Comparative study of diarrhea associated with the use of antibiotics].

BACKGROUND: Clostridium difficile is considered as one of the principal enteropathogens of nosocomial origin in developed countries in the last decade. No nosocomial outbreaks have been described in Spain. METHODS: A descriptive study of a nosocomial outbreak of diarrhea by C. difficile (DCD) which was produced during two months in a general hospital of 250 beds was carried out. The risk factors, clinical and analytical characteristics were compared with the cases of diarrhea associated to the use of antibiotics (DAA) produced during the same period. RESULTS: Of the 23 diarrheas studied, 17 (74%) corresponded to DAA and 6 (26%) to DCD, four of which were of nosocomial origin and were detected in the department of Nephrology. The incidence of DAA was 44/1000 patients who received antibiotic treatment per month and the incidence of DCD was 15.5/1000 patients/month. The patients with DAA received a mean of 1.82 antimicrobians per patient and in DCD 2.16 per patient (p: NS). The only differences found between DAA and DCD were: length of the symptoms (16.5 days in DCD, 8.5 days in DAA, p < 0.01) and the presence of blood in the feces (66.6% in DCD and 0 in DAA p < 0.01). No differences were observed in age, sex, base disease, mucous diarrhea, fever, mortality, high VSG and leucocytosis. Surveillance and control measures were effective in combating the epidemic outbreak. CONCLUSIONS: The diagnosis of diarrhea by C. difficile requires a high index of suspicion given the difficulty in differentiating the same from diarrhea associated to antibiotic use. The clinical profile and risk factors do not differ to those described in other geographic areas. The isolation and/or detection of toxins of C. difficile is recommended in patients submitted to antibiotic treatment and diarrhea of more than 72 hours of evolution.

Aged↗
PubMed1996

Comparison of four laboratory tests for diagnosis of Clostridium difficile-associated diarrhea.

Four different laboratory tests for diagnosis of Clostridium difficile-associated diarrhea were compared to determine the optimal one for management of patients with hospital-acquired diarrhea. Stool samples from 231 patients with diarrhea were tested by the following methods: culture for Clostridium difficile with subsequent determination of exotoxin production, with a toxigenic Clostridium difficile positive (TCP) result considered truly positive; enzyme immunoassay (EIA); latex agglutination test; and an immunobinding blot assay. The rates of positive results were as follows: EIA 5.5%, TCP 7.3%, latex agglutination 16.7%, and immunobinding blot assay 26.1%. Compared to the TCP results, the sensitivity and specificity were, respectively, 61 and 98% for EIA, 47 and 85% for latex agglutination, and 60 and 76% for the immunobinding blot assay. Samples from patients with > or = 6 stools/day were TCP and EIA positive in 27 and 17% of cases, respectively, whereas in patients with < 6 stools/day, these percentages decreased to 2 and 3%, respectively (p < 0.001). In hospitalized patients with > or = 6 stools/day, EIA appears to be the optimal test for diagnosis of Clostridium difficile-associated diarrhea, with a 73% positive predictive value and a 97% negative predictive value. However, in patients with < 6 stools/day, the prevalence of Clostridium difficile is low, and laboratory detection of this organism remains problematic.

Anti-Bacterial Agents↗
PubMed1998

Metronidazole may inhibit intestinal colonization with Clostridium difficile.

PURPOSE: Antibiotics suppress normal gut flora, allowing overgrowth of acquired or native Clostridium difficile, with release of toxins that cause mucosal inflammation. Oral metronidazole is used to treat antibiotic-associated colitis (pseudomembranous colitis). This study was designed to determine whether oral metronidazole, as part of preoperative bowel preparation, prevents or decreases incidence of antibiotic-associated colitis after elective colonic and rectal procedures. METHODS: Eighty-two patients (40 men) were prospectively, randomly assigned to receive one of two oral antibiotic regimens before colorectal surgery. All patients underwent mechanical bowel preparation with polyethylene glycol-electrolyte lavage solution before administration of oral antibiotics. Group 1 (n = 42) patients received three doses (1 g/dose) of neomycin and erythromycin. Group 2 (n = 40) patients received three doses (1 g/dose) of neomycin and metronidazole. Both groups received one preoperative and three postoperative doses of intravenous cefotetan (2 g/dose). Both groups had stool samples tested for C. difficile toxin in the preoperative and postoperative periods by enzyme-linked immunoabsorbent assay or by tissue culture cytotoxicity. Patients with preoperative stool studies positive for C. difficile were excluded from the study. RESULTS: Treatment groups were not different for age, gender, or surgical procedure. Mean age +/- 1 standard deviation was 67.6 +/- 13.6 (range, 34-94) years in Group 1 and 62.1 +/- 13.5 (range, 35-84) years in Group 2 (P = 0.069). Mean length of hospital stay +/- 1 standard deviation was 9.76 +/- 4.9 (range, 4-28) days for Group 1 and 8.05 +/- 2.6 (range, 3-14) days for Group 2 (P = 0.053). Five patients in Group 1 (neomycin and erythromycin) and one patient in Group 2 (neomycin and metronidazole) had positive stool studies for C. difficile. Relative risk of colonization with C. difficile in Group 1 was 4.76 times that in Group 2 (95 percent confidence interval, 0.581, 39). This difference was not statistically significant (P = 0.202). There were no significant differences in C. difficile colonization rates with respect to age, length of stay, or gender. CONCLUSIONS: This study suggests that there may be a clinical association between use of metronidazole preoperatively and inhibition of intestinal colonization by C. difficile in this patient population undergoing colonic and rectal surgery.

Adult↗
PubMed2005

Analysis of Clostridium difficile-associated diarrhea among patients hospitalized in tertiary care academic hospital.

The frequency of Clostridium difficile strains in stool samples of patients with diarrhea hospitalized in the hematology/oncology, surgery, orthopedics, transplantology ward, and emergency room of Davis Medical Center was analyzed. A total of 786 stool samples collected from patients with diarrhea and 180 samples taken from the hospital environment were cultured for C. difficile by routine methods. There were 119 strains of C. difficile isolated: 97 (12.3%) strains from patients' stools (no enteropathogen other than C. difficile was detected in these stool samples) and 22 (12.2%) strains from the hospital environment. It was confirmed that hospital environment plays an important role in transmission of C. difficile by AP-PCR and PCR ribotyping. Among 97 C. difficile strains isolated from patient' stools 25 were nontoxigenic (A-/B-), 67 were toxigenic (A+/B+), and 5 strains were toxin B-positive/toxin A-negative. Analysis of concomitant symptoms among hospitalized patients with diarrhea demonstrated significantly longer duration of diarrhea caused by nontoxigenic strains than in cases of diarrhea caused by toxigenic strains. On the other hand, among patients infected by toxigenic strains, significantly higher leukocytosis and longer duration of fever were observed. The resistance of isolated C. difficile strains to erythromycin and clindamycin indicated the possibility of transmission in the hospital strains with macrolide-lincosamide-streptogramin B resistance type.

Bacterial Proteins↗
PubMed1995

[Severe forms of pseudomembranous colitis caused by Clostridium difficile].

Clostridium difficile causes a broad spectrum of enteric diseases in humans, ranging from mild antibiotic-associated diarrhoea to more severe pseudomembranous colitis. The authors report four cases of life-threatening pseudomembranous colitis with haemodynamic changes. Infection due to Clostridium difficile should be kept in mind whenever a patient undergoing antibiotic therapy develops a symptomatology of an acute abdomen.

Abdomen, Acute↗
PubMed2001

Clostridium difficile-associated diarrhea in a VA medical center: clustering of cases, association with antibiotic usage, and impact on HIV-infected patients.

A case-control study of patients with stools assayed for Clostridium difficile toxin over a 24-month period at a Veterans Affairs hospital found that the majority of cases (70.6%) occurred in temporal clusters. Clustering was particularly evident on a designated human immunodeficiency virus (HIV) unit. Thirty-four (75.5%) of 45 HIV-infected patients with C difficile-associated diarrhea (CDAD) died during their hospitalization. Third-generation cephalosporins were the antibiotics most strongly associated with CDAD.

Adult↗
PubMed1997

Vancomycin-resistant enterococci in stool specimens submitted for Clostridium difficile cytotoxin assay.

The prevalence of, and clinical risk factors associated with, vancomycin-resistant enterococcal colonization were investigated in patients suspected of having Clostridium difficile infection. Stools submitted for C difficile cytotoxin testing were screened for vancomycin-resistant enterococci (VRE). Isolates were speciated and characterized further by antibiotic susceptibility testing, DNA fingerprinting, and DNA:DNA hybridization for detection of specific vancomycin resistance genes. Of the 79 evaluable patients identified during a 3-month period, 16.5% were VRE-positive. The VRE isolates were genetically heterogeneous, although all carried the vanA gene. DNA fingerprinting data suggest that patient-to-patient transmission occurred, implicating colonized patients as potential reservoirs for VRE transmission. A positive C difficile cytotoxin assay and diabetes mellitus were the only identifiable risk factors associated with VRE colonization. Patients at risk for C difficile infection therefore may serve as reservoirs for VRE.

Anti-Bacterial Agents↗
PubMed1998

The role of antimicrobial agents in the aetiology of Clostridium difficile-associated disease.

High Clostridium difficile disease rates were originally associated with clindamycin use, but this association has declined in recent years following the decline in the clinical use of clindamycin, and disease is now particularly associated with the use of broad-spectrum antibiotics, especially the cephalosporins. There are now sufficient reports in the literature to merit the discontinuation of the widespread use of cephalosporins, especially in the elderly, by the substitution, wherever possible, with antibiotics not associated with the promotion of C. difficile-associated disease. Such agents include benzylpenicillin, gentamicin, trimethoprim, ciprofloxacin and the ureido-penicillins. Widespread education about the proper use of antibiotics in clinical practice remains essential.

Anti-Bacterial Agents↗