Insight into the Mechanism of Lysogeny Control of phiCDKH01 Bacteriophage Infecting Clinical Isolate of Clostridioides difficile.

Clostridioides difficile is a causative agent of antibiotic-associated diarrhea as well as pseudomembranous colitis. So far, all known bacteriophages infecting these bacteria are temperate, which means that instead of prompt lysis of host cells, they can integrate into the host genome or replicate episomally. While C. difficile phages are capable of spontaneous induction and entering the lytic pathway, very little is known about the regulation of their maintenance in the state of lysogeny. In this study, we investigated the properties of a putative major repressor of the recently characterized C. difficile phiCDKH01 bacteriophage. A candidate protein belongs to the XRE family and controls the transcription of genes encoding putative phage antirepressors, known to be involved in the regulation of lytic development. Hence, the putative major phage repressor is likely to be responsible for maintenance of the lysogeny.

Antibiotic Resistance Profile of RT 027/176 Versus Other Clostridioides difficile Isolates in Silesia, Southern Poland.

Clostridioides difficile is an important health care-associated pathogen. The aim of this study was to analyze the antibiotic susceptibility of C. difficile isolates from feces of patients from 13 hospitals in Silesia, Poland. The incidence of CDI per 100.000 people in Silesia in 2018−2019 was higher than the average in Poland (39.3−38.7 vs. 30.2−29.5, respectively). The incidence doubled from 26.4 in 2020 to 55.1 in 2021. Two hundred and thirty stool samples tested positive for GDH (glutamate dehydrogenase) and toxins were cultured anaerobically for C. difficile. The isolates were characterized, typed, and tested for susceptibility to 11 antibiotics by E-test (EUCAST, 2021). The genes of toxins A/B and binary were detected by mPCR. Of 215 isolates, 166 (77.2%) were classified as RT 027 and 6 (2.8%) as related RT 176. Resistance to ciprofloxacin (96.7%), moxifloxacin (79.1%), imipenem (78.1%), penicillin (67%), and rifampicin (40.5%) was found. The ermB gene was detected in 79 (36.7%) strains. Multidrug resistance (MDR) was confirmed in 50 (23.3%) strains of RT 027 (94%). We concluded that a high prevalence of MDR among hypervirulent RT 027/176 C. difficile was found in the Silesian region of Poland, emphasizing the need to enhance regional infection control on CDI and antibiotic stewardships.

Retrospective Study of the Etiology, Laboratory Findings, and Management of Patients with Urinary Tract Infections and Urosepsis from a Urology Center in Silesia, Southern Poland Between 2017 and 2020.

BACKGROUND Recent studies have shown that up to 25% of sepsis cases originate in the urinary tract. Urosepsis can be associated with cystitis, lower urinary tract infections (UTIs), and upper UTIs and is most commonly caused by gram-negative bacteria. This retrospective study from a urology center in southern Poland, was conducted between 2017 and 2020 and aimed to investigate the causes, microbiology laboratory findings, and management in 138 patients with UTIs and urosepsis. MATERIAL AND METHODS Records of patients with UTIs with urosepsis admitted to the Urology Department of the hospital in Silesia, Poland, between 2017 and 2020 were analyzed retrospectively, and clinical and laboratory data were evaluated. RESULTS The 138 included patients were admitted to the hospital between 2017 and 2020. The median age of patients was 67 (20-94) years, and 59.9% (82/137) were men. The most common reasons for admission to the Urology Department were hydronephrosis due to dysfunction of urinary drainage in 36.5% (50/137) of patients and hydronephrosis due to urolithiasis in 22.6% (31/137) of patients. The main etiological agents responsible for the development of urosepsis were strains of Enterobacteriaceae in 85% of patients, of which 41.4% (48/116) produced extended-spectrum beta-lactamases (ESBL), accounting for 35.0% (48/137) of patients with urosepsis. In 83.3% (80/96) of patients, the pathogen cultured from the urine was identical to that cultured from the blood. CONCLUSIONS The identification of an increasing prevalence of urosepsis associated with ESBL-producing gram-negative rods from this single-center study highlights the importance of infection monitoring, rapid diagnosis, and multidisciplinary patient management.

Detection of Clostridioides difficile in hospital environment by using C diff Banana Broth™.

116 environmental samples from a 504 bed clinical hospital obtained in 2017/19 were inoculated into C diff Banana Broth™. Six C. difficile and 12 C. pefringens strains were isolated. Antibiotic-resistant Clostridium spp. dominated in hospital environment. To determine Clostridium spp. in hospital environment suitable medium like C diff Banana Broth™ should be used.

Clostridioides difficile Ribotype 027 (RT027) Outbreak Investigation Due to the Emergence of Rifampicin Resistance Using Multilocus Variable-Number Tandem Repeat Analysis (MLVA).

OBJECTIVE: The aim of this study was Clostridioides difficile outbreak investigation due to the emergence of rifampicin resistant ribotype 027 (RT 027) fecal isolates from patients of Polish tertiary care hospital between X. 2017 and II. 2018 using multilocus variable tandem repeat analysis (MLVA). MATERIALS AND METHODS: Twenty-nine C. difficile fecal isolates from patients of tertiary care hospital in Southern Poland were ribotyped and analyzed by MLVA. Multiplex PCR (mPCR) for genes encoding GDH (gluD), toxins A (tcdA)/ B (tcdB), 16S rDNA and binary toxin genes (ctdA and ctdB) was performed. The antibiotic susceptibility profile was determined by E-test. RESULTS: The A, B and binary toxins encoding genes were detected in all 29 C. difficile strains which were sensitive to metronidazole, vancomycin and were resistant to erythromycin, clindamycin, and moxifloxacin; resistance to imipenem demonstrated 97%, to rifampicin - 45% isolates. C. difficile strains could be grouped by MLVA into 5 distinct clusters, and the largest cluster II contains 16 strains. The comparison of rifampicin GM MIC of cluster II (n=16 strains) with all others (n=13) showed that strains from clusters I, III, IV and V possessed significantly (p <0.005) higher GM MIC and were more resistant to rifampicin. CONCLUSION: MLVA analysis proved transmission and recognized outbreak due to multidrug-resistant RT 027 C. difficile among patients of tertiary care hospital in Southern Poland. The reason for this is probably the widespread occurrence of spores in the hospital environment, which includes, among others, neglect of hygienic procedures and epidemic supervision. High resistance to imipenem (97%) and to rifampicin (45%) among C. difficile RT 027 Silesian isolates is threatening and requires further studies to elucidate this phenomenon.

Complete genome sequence of the newly discovered temperate Clostridioides difficile bacteriophage phiCDKH01 of the family Siphoviridae.

A temperate siphovirus, phiCDKH01, was obtained from a clinical isolate of Clostridioides difficile. The phage genome is a 45,089-bp linear double-stranded DNA molecule with an average G+C content of 28.7%. It shows low similarity to known phage genomes, except for phiCD24-1. Genomic and phylogenetic analysis revealed that phiCDKH01 is a newly discovered phage. Sixty-six putative ORFs were predicted in the genome, 37 of which code for proteins with predicted functions. The phiCDKH01 prophage was localized in the host genome. The results of this study increase our knowledge about the genetic diversity of tailed phages.

[The role of hospital environment and the hands of medical staff in the transmission of the Clostridioides (Clostridium) difficile infection]. / Rola srodowiska szpitalnego i rak personelu medycznego w szerzeniu sie zakazen Clostridioides (Clostridium) difficile.

This paper attempts to determine the role of the hospital environment in the spread of Clostridioides/Clostridium (C.) difficile infections based on a review of studies published in the medical literature and in the light of the authors' own experiences. Clostridioides/Clostridium difficile has recently attracted more and more attention, not only as an etiological factor of pseudomembranous intestinal inflammation and antibiotic associated diarrhea, but also as an etiologic factor of healthcare-associated infections (HAI) because of the possibility to survive in the hospital environment for a long time. This is caused by the production of spores, whose eradication options are limited. Clostridioides/Clostridium difficile spores are present not only on bedding of infected patients and their other belongings, but also on medical equipment and the hands of medical personnel, constituting a potential source of infection for other patients and some of the staff. The introduction of appropriate procedures for hand hygiene as well as for cleaning and disinfection of hospital surfaces makes it possible to reduce the number of spores and/or eradicate them. These procedures must be strictly followed to reduce the occurrence of spores in the hospital environment and to prevent further spread of C. difficile infections (CDI). Monitoring the presence of the C. difficile spores in a hospital environment using appropriate media (C diff Banana BrothTM) provides additional opportunities for culturing of C. difficile strains and determining ribotypes, especially hyperepidemic ones, which is extremely important from an epidemiological point of view. Med Pr. 2019;70(6):739-45.