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PubMed2000

The macrolide-lincosamide-streptogramin B resistance determinant from Clostridium difficile 630 contains two erm(B) genes.

The ErmB macrolide-lincosamide-streptogramin B (MLS) resistance determinant from Clostridium difficile 630 contains two copies of an erm(B) gene, separated by a 1.34-kb direct repeat also found in an Erm(B) determinant from Clostridium perfringens. In addition, both erm(B) genes are flanked by variants of the direct repeat sequence. This genetic arrangement is novel for an ErmB MLS resistance determinant.

Anti-Bacterial Agents↗
PubMed2005

Antibiotic-associated diarrhea and pseudomembranous colitis: are they less common with poorly absorbed antimicrobials?

Diarrhea is a well-known complication of antibiotic therapy. Rates of antibiotic-associated diarrhea (AAD) vary from 5 to 25%. Some antibiotics are more likely to cause diarrhea than others, specifically, those that are broad spectrum and those that target anaerobic flora. This paper reviews the effects of antibiotics on the fecal flora as well as host factors which contribute to AAD. Clinical features and treatment of AAD are also described. Prevention of AAD rests on wise antibiotic policies, the use of probiotics and prevention of acquisition in the hospital setting. Data from clinical trials suggest that poorly absorbed antimicrobials might have a decreased risk of causing AAD and Clostridium difficile-associated disease, as concluded from studies of antibiotics used for preoperative bowel decontamination and poorly absorbed antibiotics used for traveler's diarrhea. Controlled trials would prove this but are not yet available. Probiotics may be a good adjunct to poorly absorbed antibiotics to minimize the risk of diarrhea associated with antibiotics.

Anti-Bacterial Agents↗
PubMed2005

Activity of selected oxidizing microbicides against the spores of Clostridium difficile: relevance to environmental control.

BACKGROUND: Clostridium difficile is an increasingly common nosocomial pathogen, and its spores are resistant to common environmental surface disinfectants. Many high-level disinfectants (eg, aldehydes) are unsuitable for environmental decontamination because they need several hours of contact to be sporicidal. This study tested the potential of selected oxidative microbicides to inactivate C. difficile spores on hard surfaces in relatively short contact times at room temperature. METHODS: The spores of a clinical isolate of C. difficile were tested using disks (1 cm diameter) of brushed stainless steel in a quantitative carrier test. The spores of C. sporogenes and Bacillus subtilis, common surrogates for evaluating sporicides, were included for comparison. The clostridia were grown separately in Columbia broth (CB), and B. subtilis was grown in a 1:10 dilution of CB. Each disk received 10 microL test spores with an added soil load, and the inoculum was dried. One disk each was placed in a glass vial and overlaid with 50 microL test formulation; controls received an equivalent volume of normal saline with 0.1% Tween 80. At the end of the contact time the microbicide was neutralized, the inoculum recovered from the disks by vortexing, the eluates were membrane filtered, and the filters placed on plates of recovery medium. The colony-forming units (CFU) on the plates were recorded after 5 days of incubation. The performance criterion was > or = 6 log(10) (> or = 99.9999%) reduction in the viability titer of the spores. The microbicides tested were domestic bleach with free-chlorine (FC) levels of 1000, 3000, and 5000 mg/L; an accelerated hydrogen peroxide (AHP)-based product with 70,000 mg/L H2O2 (Virox STF); chlorine dioxide (600 mg/L FC); and acidified domestic bleach (5000 mg/L FC). RESULTS: Acidified bleach and the highest concentration of regular bleach tested could inactivate all the spores in < or = 10 minutes; Virox STF could do the same in < or = 13 minutes. Regular bleach with 3000 mg/L FC required up to 20 minutes to reduce the viability of the all the spores tested to undetectable levels; chlorine dioxide and the lowest concentration of regular bleach tested needed approximately 30 minutes for the same level of activity. CONCLUSIONS: Acidified bleach, Virox STF, and regular bleach (3000-5000 mg/L FC) could inactivate C. difficile spores on hard environmental surfaces in approximately 10 to 15 minutes under ambient conditions. All of these products are strong oxidizers and should be handled with care for protection of staff, but acidified and regular bleach with high levels of FC also release chlorine gas, which can be hazardous if inhaled by staff or patients.

Chlorine Compounds↗
PubMed2000

Antibiotics and Clostridium difficile diarrhea in the ambulatory care setting.

OBJECTIVE: The goal of this study was to determine the prevalence of Clostridium difficile diarrhea (CDD) and the risk for CDD associated with different oral antibiotics commonly used in the ambulatory care setting. METHODS: The prevalence of CDD was determined for enrollees in 4 UnitedHealth Group-affiliated health plans between January 1, 1992, and December 31, 1994. Cases were identified based on the presence of an inpatient or outpatient claim with a primary diagnosis of diarrhea, a pharmacy claim for a prescription drug used to treat CDD, or a physician or facility claim for the C. difficile toxin test, and were confirmed using full-text medical records. Within a retrospective cohort design, periods of risk for CDD were defined on the basis of duration of antibiotic therapy. To control for potential selection bias created by heterogeneous rates of C. difficile testing and to limit confounding due to multiple antibiotic exposures, we used a nested case-control design, restricting eligibility to subjects who underwent screening for C. difficile and who had been exposed to only 1 antibiotic risk period with a single antibiotic. RESULTS: The global prevalence of CDD in 358,389 ambulatory care enrollees was 12 per 100,000 person-years. In the nested case-control study, after controlling for other risk factors, 2 antibiotics demonstrated an increased association with CDD: cephalexin (odds ratio [OR] = 7.5, 95% CI = 1.8 to 34.7) and cefixime (OR = 6.4, 95% CI = 1.2 to 39.0). CONCLUSIONS: Although CDD is thought to occur primarily in hospitalized patients, it was found to be present in an ambulatory care population, but at a low frequency. In this population, it appeared to be associated with 2 cephalosporins but not with other types of antibiotics usually linked with nosocomial CDD. Because the frequency of C. difficile testing was shown to be more common with high-risk antibiotics, CDD may be underdiagnosed in the ambulatory care setting.

Adolescent↗
PubMed1997

Preliminary findings of the international typing study on Clostridium difficile. International Clostridium Difficile Study Group.

Preliminary results of the International Typing Study on Clostridium difficile indicate that there is excellent correlation between the phenotypic methods reliant on cell surface antigens for typing strains and that a larger number of phenotypes and genotypes of C. difficile exists than was previously appreciated by each group of investigators acting independently. Evidence has also emerged that some of the types described by each method are common to all typing methods, indicating that strains of the same type are found in hospitals in the United Kingdom, Belgium, the United States, and Australia.

Bacterial Typing Techniques↗
PubMed1993

Role of the laboratory in investigations of Clostridium difficile diarrhea.

The laboratory diagnosis of acute diarrhea due to Clostridium difficile can be based solely on the demonstration of either cytotoxin or enterotoxin in the stool. This and other noncultural methods, however, fail to provide isolates from which useful data can be obtained. Outbreaks can be recognized only by comparison of isolated strains. A dual approach of toxin detection and isolation is therefore recommended.

Bacterial Proteins↗
PubMed2003

In vitro activities of ramoplanin, teicoplanin, vancomycin, linezolid, bacitracin, and four other antimicrobials against intestinal anaerobic bacteria.

By using an agar dilution method, the in vitro activities of ramoplanin, teicoplanin, vancomycin, linezolid, and five other agents were determined against 300 gram-positive and 54 gram-negative strains of intestinal anaerobes. Ramoplanin was active at or=256 microg/ml. Ramoplanin displays excellent activity against C. difficile and other gram-positive enteric anaerobes, including vancomycin-resistant strains; however, it has poor activity against most gram-negative anaerobes and thus potentially has a lesser effect on the ecological balance of normal fecal flora.

Acetamides↗
PubMed1996

Macrofragment localization of the toxin A and toxin B genes of Clostridium difficile.

We report the physical mapping of the toxin A and B genes to the bacterial chromosome of Clostridium difficile ATCC 43594 by pulsed-field gel electrophoresis. Single and double digestions with restriction endonucleases NruI and SacII allowed localization of the toxin genes to a specific 577-kb fragment and estimation of genome size to be approximately 3.8 megabases. This effort represents the initial step in the construction of a physical map of the whole genome.

Chromosome Mapping↗
PubMed1998

[Clostridium difficile-associated diarrhea treated with homologous feces].

The incidence of Clostridium difficile-associated diarrhoea has increased during the last few years. Treatment with vankomycin or metronidazol is usually effective, but relapses are not uncommon. Some good results have been reported with faecal enemas, but it is a controversal form of treatment. 18 patients with C. difficile-associated diarrhoea were given homologous faeces from one healthy donor. In 17 patients faeces was instillated via a coloscope and in one patient via a gastrostoma. C. difficile toxin was detected in all patients. Three patients with severe colitis did not respond to the treatment. The remaining patients were clinically cured, and no relapses were observed. Treatment of C. difficile-associated diarrhoea with faeces appears to be an alternative in moderate cases. In our limited number of patients we observed a poor correlation between the clinical picture, the endoscopic findings and the histological findings in colon biopsies. The ethical aspects of treatment with faeces will continue to be subject to discussion.

Adult↗
PubMed2000

Clostridium difficile toxins influence hepatocyte protein synthesis through the interleukin 1 receptor.

HYPOTHESIS: Clostridium difficile toxins require interleukin 1 (IL-1) production or a functioning IL-1 receptor to elicit acute-phase protein production by murine hepatocytes. DESIGN: Experimental study. SETTING: Research laboratory at the DVA Medical Center, St Louis, Mo. CELLS STUDIED: Hepatocytes prepared from normal mice, from knockout mice deficient in IL-1 production due to loss of IL-1 converting enzyme, or from knockout mice deficient in the IL-1 p80 receptor. INTERVENTIONS: Cells were treated with lipopolysaccharide, a crude C difficile toxin extract, or purified C difficile toxins A or B for 24 hours in vitro, then radiolabeled with (35)S methionine. Newly synthesized acute-phase proteins were identified by electrophoresis and autoradiography. MAIN OUTCOME MEASURES: Synthesis of a 23-kd acute-phase protein in response to the various stimuli. RESULTS: Lipopolysaccharide, C difficile culture extract, and purified toxins A and B stimulated the synthesis of the 23-kd acute-phase protein by hepatocytes from normal mice and by hepatocytes from knockout mice deficient in the IL-1 converting enzyme. However, hepatocytes from knockout mice deficient in the IL-1 p80 receptor failed to produce this acute-phase protein when treated with the C difficile toxins, although they responded fully to lipopolysaccharide. CONCLUSIONS: Stimulation of acute-phase protein synthesis by C difficile toxins does not require IL-1 production, but does require a functioning IL-1 p80 receptor. This suggests that some of the actions of these toxins are mediated by this receptor.

Acute-Phase Proteins↗
PubMedDate not supplied

Antibiotic associated colitis.

It is a prospective study based on 100 consecutive cases of diarrhea following antibiotic therapy admitted to the pediatric services of J.N. Medical College, A.M.U., Aligarh between January to December 1987. They had C. penicillin (50), chloramphenicol (34), ampicillin (34), gentamicin (34), cephalosporin (4) and cotrimoxazole (4) for 3 days to 3 weeks prior to the onset of diarrhea. Apart from routine and special investigations, naked eye and microscopic examination of stool, its culture for pathogens including Cl. difficile were carried out in all cases. Presence of Cl. difficile cytotoxin was demonstrated by observing the cytopathic. Effect on veru cell culture, 18 grew Cl. difficile (14 cyto toxin positive). Frequency of fever, vomiting, abdominal distension, dehydration and duration of diarrhea was not different (p > 0.05) in the two groups. Purge rate and presence of mucus and blood in Cl. difficile positive patients was significantly higher (p < 0.05). Eight Cl. difficile positive (7 cytotoxin+ve) were subjected to endoscopy. Three of them showed P.M. colitis and 2 non specific colitis. Chloromycetin, gentamicin and penicillin were the main culprits responsible for AAC. None of the patients given ampicillin alone suffered from AAC. The mortality was 5%.

Anti-Bacterial Agents↗
PubMed1992

Specific binding of nucleotides and NAD+ to Clostridium difficile toxin A.

Binding of nucleotides, a tetrapolyphosphate, and NAD+ to purified toxin A of Clostridium difficile was determined by monitoring changes in intrinsic fluorescence following excitation at 280 nm, and recording emissions at 340 nm. Binding was specific for concentrations over the range 5 to 100 microM for ATP, GTP, and their respective non-hydrolysable analogues AMP-PNP and Gpp(NH)p, tetrapolyphosphate and NAD+.

Adenosine Triphosphate↗
PubMed2005

Regulation of toxin and bacteriocin synthesis in Clostridium species by a new subgroup of RNA polymerase sigma-factors.

Many Clostridium species are pathogenic for humans and animals, and most of the resulting diseases, such as tetanus, botulism, gas gangrene and pseudomembranous colitis, are due to the production of potent extracellular toxins. The biochemical mechanisms of action of Clostridium toxins have been extensively studied in the past ten years. However, detailed information about the regulation of toxin gene expression has only recently emerged. TcdR, BotR, TetR and UviA are now known to be related alternative RNA polymerase sigma factors that drive transcription of toxin A and toxin B genes in C. difficile, the neurotoxin genes in C. botulinum and C. tetani, and a bacteriocin gene in C. perfringens. Although the Clostridium sigma factors have some similarity to members of the ECF sigma factor group, they differ sufficiently in structure and function so that they have been assigned to a new group within the sigma(70)-family.

Amino Acid Sequence↗
PubMed1999

Antibacterial activity of Aristolochia paucinervis Pomel.

Several fractions of the methanolic extract of the rhizome or the leaves of Aristolochia paucinervis Pomel were screened for antibacterial activity using the agar dilution method against fourteen reference bacterial strains. Only three fractions (defatted chloroformic rhizome fraction: APRC, rhizome ethyl acetate fraction: APRE and leaf chloroform fraction: APLC) showed an activity against at least one of the microorganisms tested. The minimum inhibitory concentration (MIC) determination showed that APRC was the most active against Clostridium perfringens, Clostridium difficile, Enterococcus faecalis, Micrococcus luteus and Bacillus subtilis. The high bacteriostatic activity of APRC was confirmed by its MIC determination against clinical strains of C. perfringens (n = 32), C. difficile (n = 31), and E. faecalis (n = 22). Results of this study suggest the potential interest of this highly active fraction and support the use of A. paucinervis Pomel in Moroccan traditional medicine to treat skin and soft-tissue infections, especially gas gangrene and intestinal diseases.

Anti-Bacterial Agents↗
PubMed2003

Risk factors for Clostridium difficile colonisation and toxin production.

OBJECTIVES: to examine risk factors for patients becoming Clostridium difficile culture and toxin positive. DESIGN: prospective cohort study. SETTING: two medicine for the elderly wards. PARTICIPANTS: patients admitted to the wards over a 17-month period. MEASUREMENT: presence of Clostridium difficile on culture of stool specimens and toxins A and/or B. Patient's age, sex, source of admission, antibiotic, laxative, antacid and steroid use, presence/absence of colonic disease, neoplasia, leukaemia and nasogastric or percutaneous endoscopic gastrostomy tube feeding. RESULTS: 390 of 865 patients admitted provided a total of 1003 faecal specimens. Age (OR 1.04, 95% CI 1.001-1.07 per year), admission from another hospital (OR 2.13, 95% CI 1.29-3.50), non-cephalosporin antibiotics (OR 2.08, 95% CI 1.25-3.46) and cephalosporin use (OR 8.45, 95% CI 2.99-23.9) increased risk of becoming Clostridium difficile culture positive. Becoming toxin-positive was associated with antibiotic use only (OR 3.02, 95% CI 1.15-7.92), specifically amoxycillin (OR 8.72, 95% CI 1.66-45.9) and cephalosporins other than ceftriaxone (OR 7.28, 95% CI 1.34-39.6). CONCLUSION: different risk factors are important for the two stages leading to Clostridium difficile diarrhoea. Age, source of admission and third generation cephalosporins increase risk of becoming culture positive, whilst only antibiotic use is associated with the step of becoming toxin-positive. Understanding these differential risks may aid infection control strategies.

Aged↗
PubMed1993

Elevated levels of serum immunoglobulins in asymptomatic carriers of Clostridium difficile.

Serum levels of IgA, IgM, and polyvalent immunoglobulins reactive with Clostridium difficile were determined by ELISA for asymptomatic carriers (n = 5), symptomatic individuals (n = 21), and a pool of 30 "normal" individuals. Mean IgA concentrations expressed as optical density (OD +/- SD) were significantly higher (P < .001) for asymptomatic carriers (1.252 +/- 0.516) than for symptomatic patients (0.374 +/- 0.145). Mean serum IgM levels also were significantly higher (P < .001) for carriers (1.456 +/- 0.582) than for symptomatic patients (0.727 +/- 0.331), as were mean values for polyvalent immunoglobulins (2.25 +/- 0.718 for carriers vs. 1.457 +/- 0.574 for symptomatic patients; P < .05). Although the patient populations were small, the levels of immunoglobulins reactive with C. difficile antigens differed significantly. This difference might reflect the ability of the host to mount an immune response and might be a factor influencing whether a patient develops disease due to this organism. The ability to detect differences in immunoglobulin levels might also help differentiate infection from colonization.

Antibodies, Bacterial↗
PubMed2005

Subtyping of Clostridium difficile PCR ribotype 001 by REP-PCR and PFGE.

The REP-PCR (repetitive sequence-based PCR using repetitive extragenic palindromic primers) typing method and a modified PFGE method were applied to isolates of Clostridium difficile PCR ribotype 001 with the aim of comparing their performance as methods of subtyping this organism. Of 200 isolates from 60 hospitals tested by REP-PCR, eight subtypes were identified and labelled as REP-PCR subtypes 001-008. The predominant subtype, REP-PCR subtype 003, accounted for 47% of the total. Fifty-two of the 200 isolates were analysed by a modified PFGE method and seven subtypes were identified, labelled as PF-A-PF-G. There was excellent correlation between REP-PCR subtypes and PFGE subtypes with both methods displaying broadly similar discriminatory powers. However, REP-PCR subtyping proved to be a much easier, cheaper and more rapid method suitable for application for routine subtyping of C. difficile ribotype 001. Application of REP-PCR subtyping to UK isolates of C. difficile PCR ribotype 001 from 60 different centres revealed a wide distribution of REP-PCR subtype 003 throughout England and Wales, with a regional clustering of REP-PCR subtype 001 around Northwest England and North Wales. Analysis of isolates from a single hospital over a 4-year period revealed a change in predominant subtype over time.

Clostridioides difficile↗
PubMed1991

Prospective study of Clostridium difficile intestinal colonization and disease following single-dose antibiotic prophylaxis in surgery.

A total of 108 volunteers undergoing an elective surgical procedure were randomly given a single 2-g intravenous prophylactic dose of either a cephalosporin or mezlocillin. Stool samples were cultured for Clostridium difficile the day before the operation and later on postoperative days 4, 7, and 14. C. difficile was detected in 23.0% of patients who received a cephalosporin (cefoxitin, 8.3%; cefazolin, 14.3%; cefotetan, 20.0%; ceftriaxone, 25.0%; cefoperazone, 43.7%), in 3.3% of patients given mezlocillin, and in none of 15 control volunteers given no antimicrobial agent. No patient experienced diarrhea.

Adolescent↗