•Between October and December 2016, 36 Slovak hospitals performed Clostridium difficile infection (CDI) surveillance.•The overall mean CDI incidence was 2.8 (95% confidence interval 1.9–3.9) cases ...per 10 000 patient-days.•The most prevalent ribotypes were 001 (n=46, 59.0%) and 176 (n=23, 29.5%).•Almost all isolates showed reduced susceptibility to moxifloxacin (n=73, 93.6%).•A reduced susceptibility to metronidazole was observed in 13 primary fresh isolates.
To obtain standardized epidemiological data for Clostridium difficile infection (CDI) in Slovakia.
Between October and December 2016, 36 hospitals in Slovakia used the European Centre for Disease Prevention and Control (ECDC) Clostridium difficile infection (CDI) surveillance protocol.
The overall mean CDI incidence density was 2.8 (95% confidence interval 1.9–3.9) cases per 10 000 patient-days. Of 332 CDI cases, 273 (84.9%) were healthcare-associated, 45 (15.1%) were community-associated, and 14 (4.2%) were cases of recurrent CDI. A complicated course of CDI was reported in 14.8% of cases (n=51). CDI outcome data were available for 95.5% of cases (n=317). Of the 35 patients (11.1%) who died, 34 did so within 30 days after their CDI diagnosis.
Of the 78 isolates obtained from 12 hospitals, 46 belonged to PCR ribotype 001 (59.0%; 11 hospitals) and 23 belonged to ribotype 176 (29.5%; six hospitals). A total of 73 isolates (93.6%) showed reduced susceptibility to moxifloxacin (ribotypes 001 and 176; p< 0.01). A reduced susceptibility to metronidazole was observed in 13 isolates that subsequently proved to be metronidazole-susceptible when, after thawing, they were retested using the agar dilution method. No reduced susceptibility to vancomycin was found.
These results show the emergence of C. difficile ribotypes 027 and 176 with a predominance of ribotype 001 in Slovakia in 2016. Given that an almost homogeneous reduced susceptibility to moxifloxacin was detected in C. difficile isolates, this stresses the importance of reducing fluoroquinolone prescriptions in Slovak healthcare settings.
We reviewed the molecular epidemiology of Clostridioides difficile infection (CDI) in Japan by reviewing articles in which typing analysis was performed on recovered C. difficile isolates. Most of ...the multicenter studies showed that the major prevalent PCR-ribotypes (RT018-related type, RT014, RT002, RT369, and RT017) accounted for more than 75% of clinical isolates in Japan, which has not changed significantly since the late 1990s. Within the RT018-related isolates, a shift from RT018 to RT018ʹʹ (QX239) and the persistence of high levels of antimicrobial resistance were observed. Among toxin A-negative, toxin B-positive C. difficile, RT017 was replaced by RT369, which was more resistant to fluoroquinolone. The isolation rate of binary toxin-positive isolates was low (2–6%), except in one study (10%). Isolation of RT027 and RT078 was rare in endemic settings, while there was a first report of a nosocomial outbreak due to RT027 C. difficile in 2019. Notably, the vast majority of RT027 isolates, including the epidemic strain responsible for the outbreak, were susceptible to moxifloxacin, suggesting that Japanese RT027 represents the pre-epidemic RT027 genetic background. To understand the CDI burden in Japan, a nationwide strain-based surveillance system is imperative.
•C.difficile prevalent in Japan are RT018-related, RT014, RT002, RT369, and RT017.•The prevalence of the major ribotypes has not significantly changed for decades.•Within RT018-related type, a shift from RT018 to RT018ʹʹ (QX239) has occurred.•In A−B+ isolates, RT017 was replaced by RT369 with fluoroquinolone resistance.
Antibiotics are frequently used in feed to control bacterial diseases in aquaculture settings. In addition to the intended purpose of controlling diseases, applications of medicated feed may ...significantly change the composition of the gut microbiota, which in turn may have impact on host-pathogen interactions. However, nothing is known concerning the effects of antibiotics on microbiota in the gut channel catfish (Ictalurus punctatus), the most important aquaculture species in the United States. In this study, we determined the impact of medicated feed containing florfenicol on microbiota in the gut of catfish at various times post-feeding. Through analysis of bacterial 16S rRNA gene amplicons, we found that the medicated feed had a dramatic impact on the composition of the gut microbiota, with significant enrichment of Plesiomonas spp., accounting for 66% of all gut bacteria. Along with other related species, Proteobacteria taxa accounted for 93% of all microbiota by day 10; in contrast, the gut microbiota of fish receiving non-medicated feed harbored a bacterial assemblage that had greater ribotype richness and a more even distribution (P < .05), with <10% Proteobacteria relative abundance. Moreover, florfenicol-medicated feed resulted in an increased relative abundance of potential opportunistic pathogens including Plesiomonas and Aeromonas species, which may have detrimental impacts on fish health. This study provided insights into the specific bacterial taxa within the channel catfish intestinal microbiome that were impacted by florfenicol-medicated feed and suggests that the abuse of this and other antibiotics by the aquaculture industry may induce the relative abundance of potentially pathogenic gut microorganisms.
•Florfenicol-medicated feed had a dramatic impact on catfish gut microbiota.•Florfenicol-medicated feed reduced the composition and diversity of gut microbiota.•Florfenicol-medicated feed resulted in an increasing of potential aquatic pathogens.•Catfish with non-medicated feed harbored more rich gut bacterial assemblages.
Clostridioides difficile biofilms are believed to protect the pathogen from antibiotics, in addition to potentially contributing to recurrent infections.
Biofilm production of 102 C. difficile ...isolates was determined using the crystal violet staining technique, and detachment assays were performed. The expression levels of cwp84 and slpA genes were evaluated by real-time PCR on selected isolates.
More than 70% of isolates (75/102) were strong biofilm producers, and the highest detachment of biofilm was achieved with the proteinase K treatment (>90%). The overall mean expression of cwp84 was higher in RT027 than in RT001 (p=0.003); among strong biofilm-producing strains, the slpA expression was lower in RT027 than in RT001 (p<0.000).
Proteins seem to have an important role in the biofilm's initial adherence and maturation. slpA and cwp84 are differentially expressed by C. difficile ribotype and biofilm production level.
Se cree que las biopelículas de Clostridioides difficile (C. difficile) protegen al patógeno de los antibióticos, además de contribuir potencialmente a las infecciones recurrentes.
Se determinó la producción de biopelículas de 102 aislados de C. difficile, mediante la técnica de tinción con violeta cristal y se realizaron ensayos de desprendimiento. Los niveles de expresión de los genes cwp84 y slpA se evaluaron mediante PCR en tiempo real en aislados seleccionados.
Más del 70% de los aislados (75/102) fueron fuertes productores de biopelículas y el mayor desprendimiento de biopelícula se logró con el tratamiento con proteinasa K (> 90%). La expresión media global de cwp84 fue mayor en RT027 que en RT001 (p = 0,003); entre las cepas productoras fuertes de biopelícula, la expresión de slpA fue menor en RT027 que en RT001 (p < 0,000).
Las proteínas parecen tener un papel importante en la adhesión y maduración inicial de las biopelículas; slpA y cwp84 se expresan de forma diferente según el ribotipo de C. difficile y el nivel de producción de biopelículas.
While Clostridium difficile epidemiology is well documented in many European countries, data are largely missing for South Eastern European region. Here we report the PCR ribotype distribution of 249 ...C. difficile isolates received for typing from six hospital settings from Croatia, Bosnia and Herzegovina, Republic of Macedonia and Serbia in time period from 2008 to 2015. Twenty-four PCR ribotypes were detected. The majority of strains from Bosnia and Herzegovina and Serbia belonged to PCR ribotype 027 (65.8%). Other three dominating PCR ribotypes were 176 (18 strains; Croatia), 001/072 (15 strains; all countries) and 014/020 (15 strains; all countries).
•Ribotype distribution for C. difficile strains from selected hospitals from four South East European countries is given.•In epidemic settings PCR ribotypes 027 and 176 were dominant.•In non-epidemic settings PCR ribotypes 001/072 and 014/020 were dominant.
Patients with cancer are particularly vulnerable to Clostridioides difficile infection (CDI). Guidelines recommend a two-step diagnostic algorithm to differentiate carriers from CDI; however, there ...are limited data for this approach while including other confounding risk factors for diarrhea such as radiation, cytotoxic chemotherapy, and adoptive cell based therapies.
We conducted a prospective, non-interventional, single center, cohort study of cancer patients with acute diarrhea and C. difficile, identified in stools by nucleic acid amplification tests (NAAT) and culture. Fecal toxin A/B was detected by enzyme immunoassay (EIA) and isolates were ribotyped using 16s rRNA fluorescent sequencing. Patients were followed for 90 days to compare outcomes according to malignancy type, infecting ribotype, and EIA status.
We followed 227 patients with a positive NAAT. Of these, 87% were hospitalized and 83% had an active malignancy. EIA was confirmed positive in 80/227 (35%) of patients. Those with EIA+ were older (60 ± 18 years vs 54 ± 19 years., P = .01), more likely to fail therapy 24/80 (30%) vs 26/147 (18%), P = .04 and experience recurrence 20/80 (25%) vs 21/147(14%), P < .05. We found a low prevalence (22%) of ribotypes historically associated with poor outcomes (002, 018, 027, 56, F078-126, 244) but their presence were associated with treatment failure 17/50 (34%) vs 33/177 (19%), P = .02.
When compared to cancer patients with fecal NAAT+/EIA-, patients with NAAT+/EIA+ CDI are less likely to respond to therapy and more likely to experience recurrence, particularly when due to ribotypes associated with poor outcomes.
The epidemiological landscape of Clostridium difficile infection (CDI) has changed over the past 30 years.
To review studies of CDI in the community setting.
Electronic databases including PubMed, ...MEDLINE, Embase, Google Scholar, Scopus, ClinicalTrials.gov and Cochrane Databases were searched for human studies performed between 2000 and 2017 that assessed the epidemiology, risk factors, ribotypes, hospital and intensive care unit (ICU) outcomes, and management of community-acquired CDI. In addition, references were searched manually to identify other relevant studies.
In total, 39 articles met the inclusion criteria. The incidence of community-acquired CDI has almost doubled in the past decade. Approximately half of all cases of CDI are attributed to community origin. Individuals who are younger, female, in the presence of infants, frequently use proton pump inhibitors or specific classes of antibiotics, or live near farms and livestock are at higher risk for community-acquired CDI. Additionally, approximately 40% of all community-acquired cases require hospitalization, where severity has been linked to hypervirulent ribotypes 027 and 078 with poor outcomes. Emerging data on treatment paradigms have led to the revision of clinical guidelines and two potential vaccines in phase three clinical trials. However, ribotype-specific responses to current treatment strategies are lacking.
Community-acquired CDI represents a growing public health threat and burden on healthcare systems. A multi-disciplinary approach will be required to stem the tides.
Three patients with severe Clostridium difficile infection (CDI) caused by an unusual strain of C. difficile, PCR ribotype (RT) 251, were identified in New South Wales, Australia. All cases presented ...with severe diarrhoea, two had multiple recurrences and one died following a colectomy. C. difficile RT251 strains were isolated by toxigenic culture. Genetic characterisation was performed using techniques including toxin gene profiling, PCR ribotyping, whole genome sequencing (WGS), in-silico multi-locus-sequence-typing (MLST) and core-genome single nucleotide variant (SNV) analyses. Antimicrobial susceptibility was determined using an agar incorporation method. In vitro toxin production was confirmed by Vero cell cytotoxicity assay and pathogenicity was assessed in a murine model of CDI. All RT251 isolates contained toxin A (tcdA), toxin B (tcdB) and binary toxin (cdtA and cdtB) genes. Core-genome analyses revealed the RT251 strains were clonal, with 0–5 SNVs between isolates. WGS and MLST clustered RT251 in the same evolutionary clade (clade 2) as RT027. Despite comparatively lower levels of in vitro toxin production, in the murine model RT251 infection resembled RT027 infection. Mice showed marked weight loss, severe disease within 48 h post-infection and death. All isolates were susceptible to metronidazole and vancomycin. Our observations suggest C. difficile RT251 causes severe disease and emphasise the importance of ongoing surveillance for new and emerging strains of C. difficile with enhanced virulence.
•Three cases of severe Clostridium difficile infection caused by Ribotype 251.•Mouse model confirms RT251 causes a rapidly fatal disease comparable to RT027.•RT251 belongs to the same evolutionary clade as RT027.•Arrival of RT251 in Australia possibly through imported food products.
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