Search

Purpose: To characterise and compare twenty-eight Finnish Clostridium difficile RT027-like isolates, selected based on the presence of 18 bp deletion in the tcdC gene and toxin gene profile (A, B, binary), with eleven RT027 isolates from different Finnish geographical areas and time periods., Methods: Twenty-eight C. difficile RT027-like isolates and 11 RT027 comparative strains were characterised by capillary-electrophoresis (CE) ribotyping, multi-locus variable tandem-repeats analysis (MLVA), multi-locus sequence typing (MLST), and sequencing of tcdC and gyrA gene fragments. Susceptibility to moxifloxacin was determined by E-test., Results: Of 28 RT027-like isolates, seven RTs (016, 034, 075, 080, 153, 176 and 328), three WEBRIBO types (411, 475, AI-78) and three new profiles (F1-F3) were identified. MLVA revealed six clonal complexes (RTs 016, 027, 176 and F3). MLST showed eleven sequence types (1, 41, 47, 67, 95, 191,192, 223, 229, 264 and new ST). Twenty-two isolates (RTs 016, 080, 176, 328, F1, F2, F3 and WRTAI-78) carried Δ117 in the tcdC gene. Isolates of RTs 016, 027 and 176 were moxifloxacin resistant and harboured Thr82Ile in the GyrA., Conclusion: Our results show a high diversity within 28 Finnish RT027-like C. difficile isolates, with twelve CE-ribotyping profiles and eleven STs. MLVA revealed the regional spread of RTs 016, 027, 176 and F3. The presence of Δ117 in the tcdC gene in eight non-027 RTs highlights the importance of careful interpretation of the results from molecular systems targeting this site in the genome of C. difficile and the need of strain typing for epidemiological purposes., (Copyright © 2017. Published by Elsevier B.V.)

This study aimed to characterize Clostridium difficile isolates cultured from stool samples of patients with C. difficile infection (CDI) and swabs from a medical environment in a gastroenterology center in Tehran, Iran. A total of 158 samples (105 stool samples from hospitalized patients and 53 swabs from medical devices and the environment) were collected from January 2011 to August 2011 and investigated for the presence of C. difficile by direct anaerobic culture on a selective media for C. difficile. C. difficile isolates were further characterized by capillary electrophoresis (CE) ribotyping and toxin gene multiplex PCR. Of 158 samples, C. difficile was cultured in 19 of 105 stool samples (18%) and in 4 of 53 swabs (7.5%). C. difficile PCR ribotype (RT) 126 was the most common RT in the study (21.7%). Further RTs were: 001, 003, 014, 017, 029, 039, 081, 103 and 150. RTs 126, 001, 150 were cultured from both the stool samples and swabs of medical devices and the hospital environment which suggest a possible route of transmission., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

In 2014, 18 hospitals in the Czech Republic participated in a survey of the incidence of Clostridium difficile infections (CDI) in the country. The mean CDI incidence was 6.1 (standard deviation (SD):7.2) cases per 10,000 patient bed-days and 37.8 cases (SD: 41.4) per 10,000 admissions. The mean CDI testing frequency was 39.5 tests (SD: 25.4) per 10,000 patient bed-days and 255.8 tests (SD: 164.0) per 10,000 admissions. A total of 774 C. difficile isolates were investigated, of which 225 (29%) belonged to PCR ribotype 176, and 184 isolates (24%) belonged to PCR ribotype 001. Multilocus variable-number tandem repeat analysis (MLVA) revealed 27 clonal complexes formed by 84% (190/225) of PCR ribotype 176 isolates, and 14 clonal complexes formed by 77% (141/184) of PCR ribotype 001 isolates. Clonal clusters of PCR ribotypes 176 and 001 were observed in 11 and 7 hospitals, respectively. Our data demonstrate the spread of two C. difficile PCR ribotypes within 18 hospitals in the Czech Republic, stressing the importance of standardising CDI testing protocols and implementing mandatory CDI surveillance in the country., (This article is copyright of The Authors, 2016.)

Background: Clostridium difficile infection (CDI) is a major cause of antibiotic-associated diarrhoea. Given an increasing CDI incidence and global spread of epidemic ribotypes, a 1-year study was performed to analyse the molecular characteristics of C. difficile isolates and associated clinical outcomes from patients diagnosed with CDI in the Internal Medicine department at University Hospital Motol, Prague from February 2013 to February 2014., Results: A total of 85 unformed stool samples were analysed and CDI was laboratory confirmed in 30 patients (6.8 CDI cases per 10,000 patient bed days and 50.6 CDI cases per 10,000 admissions). The CDI recurrence rate within 3 months of treatment discontinuation was 13.3% (4/30). Mortality within 3 months after first CDI episode was 26.7% (8/30), with CDI the cause of death in two cases. 51.9% of C. difficile isolates belonged to PCR-ribotype 176. MLVA of ribotype 176 isolates revealed two clonal complexes formed by 10/14 isolates. ATLAS scores and Horn's index were higher in patients with ribotype 176 infections than with non-ribotype 176 infections., Conclusion: This study highlights the clinical relevance of C. difficile PCR-ribotype 176 and its capacity to spread within a healthcare facility.

The first case of Clostridium difficile RT027 infection in the Czech Republic (CZ) was identified. The patient had been hospitalised in Germany prior to moving to CZ. Multiple-Locus Variable number tandem repeat Analysis revealed a genetic relatedness between the patient's isolate and RT027 isolate collected in the German hospital., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Clostridium difficile has been recovered from the faeces of several animal species as well as horses. Between April 2015 and October 2016, 213 samples of faeces from non-hospitalized (n = 138) and hospitalized horses (n = 75) were investigated and eighteen C. difficile isolates were cultured using an enrichment method. Sixteen C. difficile positive samples were identified from hospitalised horses (p < 0.01). Molecular typing revealed seven ribotypes and sequence types (RT033/ST11 n = 8, 44.4%; RT081/ST9 n = 4, 22.2%; RT009/ST3 n = 2, 11.1%; RT003/ST12 n = 1, 5.6%; RT010/ST15 n = 1, 5.6%; RT012/ST54 n = 1, 5.6%; RT039/ST26 n = 1, 5.6%). Seven identified STs clustered to two clades (1 and 5). All C. difficile isolates were susceptible to amoxicillin, metronidazole, moxifloxacin, and vancomycin. One isolate (RT039) exhibited a high level of resistance to erythromycin and clindamycin (256 mg/L) and carried the ermB , adenine methylase gene. Five isolates were resistant to clindamycin at lower minimal inhibitory concentrations (MICs = 8-16 mg/L) and were susceptible to erythromycin and also ermB negative. All isolates were resistant to enrofloxacin (MICs ranged between 4 and 32 mg/L). Eight isolates were resistant to tetracycline (MICs 12-32 mg/L). Of them, four isolates carried the tetM gene and four isolates the tetW gene. In addition, the tetracycline resistance determinants identified were: tetA (P) (n = 4); tetB (P); and tetL (n = 1 each). The presence of tetW or tetM , together with other tet- class mechanisms, lead to an increase in the MICs to tetracycline. C. difficile isolates derived from Czech horses are identical to the ribotypes identified in humans and carry acquired antimicrobial resistance genes whose dissemination from veterinary healthcare sector to humans should be monitored by the „One health" approach. • A prevalence of C. difficile 8.4% (18/213) was identified in Czech horses. • C. difficile isolates belonged to seven ribotypes and sequence types; ribotype 033 predominates. • All C. difficile horse isolates were resistant to enrofloxacin. • 50% of isolates were resistant to tetracycline and carried the tetM or tetW genes. • 33.3% of isolates were resistant to clindamycin and one also to erythromycin. [ABSTRACT FROM AUTHOR]

Purpose The objective of this survey was to determine the incidence of Clostridium difficile infections (CDI) at the Department of Infectious Diseases, Bulovka Hospital, and to evaluate clinical and epidemiological data on CDI patients together with a detailed molecular characterisation of C . difficile isolates. The patient outcomes were correlated to causative C . difficile PCR-ribotype. Methods The twelve-month study (2013) comprised patients two years of age and older with CDI. CDI severity was estimated using ESCMID criteria and ATLAS scoring. C . difficile isolates were further characterized using ribotyping, Multiple-Locus Variable Tandem-Repeats analysis (MLVA) and investigation of antibiotic-resistance determinants ( gyrA , gyrB , rpoB , ermB ). Results A total of 619 diarrhoeal stools were investigated. Seventy-two stool samples were GDH and toxin A/B positive, and 39 samples were GDH positive only and subsequently toxigenic C . difficile was cultured. In total, 111 C . difficile isolates were characterized, of which 64 (57.7%) belonged to PCR-ribotype 176. MLVA analysis of PCR-ribotype 176 isolates revealed 11 clonal complexes. Seventy-two isolates (64.9%) showed amino acid substitution Thr82Ile in the GyrA , and sixty-two isolates (55.9%) showed amino acid substitutions Arg505Lys together with His502Asn, or Asp492Glu together with Arg505Lys in the RpoB . Twelve isolates (10.8%) were ermB positive. Severe CDI according to the ESCMID criteria was recorded in forty-two patients (37.8%), and sixteen patients (14.4%) had ATLAS score ≥ 6. Twenty-nine patients (26.1%) had recurrent CDI and twenty-four patients (21.6%) died during the study period. Conclusions A higher rate of severe CDI, recurrences and mortality in association with PCR-ribotype 176 infections were observed. The high incidence of PCR-ribotype 176 in the study, and the presence of clonal relatedness between PCR-ribotype 176 isolates, indicate its higher capacity to spread in a hospital setting, which in turn highlights the need to implement strict epidemic measures when PCR-ribotype 176 occurs. [ABSTRACT FROM AUTHOR]