Clostridium perfringens – antibiotic-associated and sporadic diarrhea in children: age, anamnestic and clinical and laboratory features

The article presents the data of the research on the problem of two clinical forms of enteroproducing Clostridium perfringens (C. perfringens) – associated infection in the children who are not connected with food – antibiotic-associated and sporadic forms. Within the period of 2019–2021 a retrospective analysis of the results of complex clinical and laboratory study of children aged from 2 months to 18 years with diarrhea (acute intestinal infection, n = 406 children) was carried out within the framework of the algorithm developed by Pediatric Research and Clinical Center for Infectious Diseases (Russia, Saint Petersburg). The material for the study included stool tests, coprofiltrates and strains of the microorganisms determinating microbiocoenosis of the intestines including enterotoxigenic strains of C. perfringens. The main group included children with diarrhea (n = 38 of 406 children with diarrhea) who at admission presented C. perfringens enterotoxin in coprofiltrates found by immune-enzyme analysis and there were identified the cultures of enterotoxigenic strains of C. perfringens by tests of intestinal contents. The first group included children (n = 30 of 406 children; 7 %) with antibiotic-associated diarrhea (AAD). The second group included children (n = 8 of 406 children; 2 %) with sporadic diarrhea. An overwhelming majority of patients with ААD (n = 20; 92.3 %) of the first 8 (100 %) years of life presented intensity of diarrhea syndrome in direct dependence on colonization activity of enterotoxigenic C perfringens (colony-forming units/g feces) (r = 0.78). The disease proceeded mainly with the syndromes of gastroenteritis, enterocolitis and hemorrhagic colitis. There was noted a reliable high frequency of severe forms of both ААD, and sporadic diarrhea (р < 0.05) caused by enterotoxigenic strains of C perfringens in children of the first year of life. Deep decompensated disorders of large intestine microbiota associated with Klebsiella spp., Staphylococcus aureus and Candida albicans, were characterized by severity and prolonged character of the course (р < 0.05). The received results determine the tactics of differential diagnosis and treatment of AAD and sporadic diarrhea caused by enteroproducing strains of C. perfringens.

Cereus sinensis Polysaccharide Alleviates Antibiotic-Associated Diarrhea Based on Modulating the Gut Microbiota in C57BL/6 Mice

The present study investigated whether the purified polysaccharide from Cereus sinensis (CSP-1) had beneficial effects on mice with antibiotic-associated diarrhea (AAD). The effects of CSP-1 on gut microbiota were evaluated by 16S rRNA high-throughput sequencing. Results showed that CSP-1 increased the diversity and richness of gut microbiota. CSP-1 enriched Phasecolarctobacterium, Bifidobacterium and reduced the abundance of Parabacteroides, Sutterella, Coprobacillus to near normal levels, modifying the gut microbial community. Microbial metabolites were further analyzed by gas chromatography-mass spectrometry (GC-MS). Results indicated CSP-1 promoted the production of various short-chain fatty acids (SCFAs) and significantly improved intestinal microflora dysfunction in AAD mice. In addition, enzyme linked immunosorbent assay and hematoxylin-eosin staining were used to assess the effects of CSP-1 on cytokine levels and intestinal tissue in AAD mice. Results demonstrated that CSP-1 inhibited the secretion of interleukin-2 (IL-2), interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) and improved the intestinal barrier. Correspondingly, the daily records also showed that CSP-1 promoted recovery of diarrhea status score, water intake and body weight in mice with AAD. In short, CSP-1 helped alleviate AAD by regulating the inflammatory cytokines, altering the composition and richness of intestinal flora, promoting the production of SCFAs, improving the intestinal barrier as well as reversing the dysregulated microbiota function.

A worldwide systematic review and meta-analysis of bacteria related to antibiotic-associated diarrhea in hospitalized patients

Introduction Antibiotic-associated diarrhea (AAD) is a major hospital problem and a common adverse effect of antibiotic treatment. The aim of this study was to investigate the prevalence of the most important bacteria that cause AAD in hospitalized patients. Materials and methods PubMed, Web of Science and Scopus databases were searched using multiple relevant keywords and screening carried out based on inclusion/exclusion criteria from March 2001 to October 2021. The random-effects model was used to conduct the meta-analysis. Results Of the 7,377 identified articles, 56 met the inclusion criteria. Pooling all studies, the prevalence of Clostridioides (Clostridium) difficile, Clostridium perfringens, Klebsiella oxytoca, and Staphylococcus aureus as AAD-related bacteria among hospitalized patients were 19.6%, 14.9%, 27%, and 5.2%, respectively. The prevalence of all four bacteria was higher in Europe compared to other continents. The highest resistance of C. difficile was estimated to ciprofloxacin and the lowest resistances were reported to chloramphenicol, vancomycin, and metronidazole. There was no or little data on antibiotic resistance of other bacteria. Conclusions The results of this study emphasize the need for a surveillance program, as well as timely public and hospital health measures in order to control and treat AAD infections.

Effects of Bacteroides-Based Microecologics against Antibiotic-Associated Diarrhea in Mice

Antibiotic-associated diarrhea (AAD) is a self-limiting disease mediated by antibiotic therapy. In clinical practice, several types of probiotics are used in treating AAD, but minimal research has been done on Bacteroides-based microecologics. Our aim was to evaluate the therapeutic effects of Bacteroidetes uniformis FGDLZ48B1, B. intestinalis FJSWX61K18, Bifidobacterium adolescentis FHNFQ48M5, and B. bifidum FGZ30MM3 and their mixture on AAD in mice. The lincomycin hydrochloride-induced AAD models were gavaged with a single strain or a probiotic mixture for a short period to assess the changes in colonic histopathology and cytokine concentrations, intestinal epithelial permeability and integrity, short-chain fatty acids (SCFAs), and the diversity of intestinal microbiota. Our data indicated that both the sole use of Bacteroides and the combination of Bacteroides and Bifidobacterium beneficially weakened systemic inflammation, increased the recovery rate of tissue structures, increased the concentrations of SCFAs, and restored the gut microbiota. Moreover, the probiotic mixture was more effective than the single strain. Specifically, B. uniformis FGDLZ48B1 combined with the B. adolescentis FHNFQ48M5 group was more effective in alleviating the pathological features of the colon, downregulating the concentrations of interleukin (IL)-6, and upregulating the expression of occludin. In summary, our research suggests that administration of a mixture of B. uniformis FGDLZ48B1 and B. adolescentis FHNFQ48M5 is an effective approach for treating AAD.

INFLUENCE OF IRRATIONAL PRESCRIPTION OF ANTIBACTERIAL THERAPY ON THE PROGNOSIS OF TREATMENT AND SURVIVAL IN PATIENTS WITH COVID-19

IIntroduction. The COVID-19 pandemic became a major challenge for healthcare systems around the world. The development and improvement of basic treatments for coronavirus patients is important to improve public health and improve quality of life after recovery. The aim of the study: to determine the frequency and structure of prescribing antibacterial drugs in the prehospital and hospital stages, used in patients with COVID-19. Assess the relationship between irrational use of antibacterial drugs with the length of hospital stay of patients with coronavirus disease, the risk of transfer to the intensive care unit (ICU) and mortality. Materials and methods: Statistical, retrospective analysis of 400 case histories of patients with COVID-19 who were treated at the Municipal Non-Profit Enterprise «Kyiv City Clinical Hospital №17» (KNP «KMKL#17») for the period from September 2020 to November 2021 with severe coronavirus disease. Results: 400 medical charts were selected for the study, which were divided into two groups according to the purpose of antibacterial therapy. Of the group of patients who received pre-hospital antibacterial therapy (200 people), indications for its appointment had only 7 % of patients. Among the group receiving antibacterial drugs there is a prolongation of the length of stay in the hospital, the risk of transfer to ICU increases. There is also higher risk of mortality in patients of group 1 (14,5 %), compared with group 2 (8 %), whose antibacterial drugs were not prescribed at the prehospital stage. Conclusion: as a result of the study it was found that patients who were unreasonably prescribed antibacterial therapy prolongs the period of general hospitalization by 2.3 ± 0.8 days, increasing the need for transfer of patients due to deterioration to ICU by an average of 13 %, increase in the incidence of antibiotic-associated diarrhea by 7-8 %, and there is a tendency to increase mortality from COVID-19. Antibacterial drugs should be used only on the basis of indications in the case of proven bacterial co-infection (superinfection) or reasonable suspicion of it in patients with respiratory disease caused by SARS-CoV-2 and in no case should be prophylactic.

An aniline-substituted bile salt analog protects both mice and hamsters from multiple Clostridioides difficile strains

Clostridioides difficile infection (CDI) is the major identifiable cause of antibiotic-associated diarrhea. The emergence of hypervirulent C. difficile strains has led to increases in both hospital- and community-acquired CDI. Furthermore, CDI relapse from hypervirulent strains can reach up to 25%. Thus, standard treatments are rendered less effective, making new methods of prevention and treatment more critical. Previously, the bile salt analog CamSA was shown to inhibit spore germination in vitro and protect mice and hamsters from C. difficile strain 630. Here, we show that CamSA was less active at preventing spore germination of other C. difficile ribotypes, including the hypervirulent strain R20291. Strain-specific in vitro germination activity of CamSA correlated with its ability to prevent CDI in mice. Additional bile salt analogs were screened for in vitro germination inhibition activity against strain R20291, and the most active compounds were tested against other strains. An aniline-substituted bile salt analog, (CaPA), was found to be a better anti-germinant than CamSA against eight different C. difficile strains. In addition, CaPA was capable of reducing, delaying, or preventing murine CDI signs in all strains tested. CaPA-treated mice showed no obvious toxicity and showed minor effects on their gut microbiome. CaPA’s efficacy was further confirmed by its ability to prevent CDI in hamsters infected with strain 630. These data suggest that C. difficile spores respond to germination inhibitors in a strain-dependent manner. However, careful screening can identify anti-germinants with broad CDI prophylaxis activity.

Production of p-cresol by Decarboxylation of p-HPA by All Five Lineages of Clostridioides difficile Provides a Growth Advantage

Clostridioides difficile is the leading cause of antibiotic-associated diarrhea and is capable of causing severe symptoms, such as pseudomembranous colitis and toxic megacolon. An unusual feature of C. difficile is the distinctive production of high levels of the antimicrobial compound para-cresol. p-Cresol production provides C. difficile with a competitive colonization advantage over gut commensal species, in particular, Gram-negative species. p-Cresol is produced by the conversion of para-hydroxyphenylacetic acid (p-HPA) via the actions of HpdBCA decarboxylase coded by the hpdBCA operon. Host cells and certain bacterial species produce p-HPA; however, the effects of p-HPA on the viability of C. difficile and other gut microbiota are unknown. Here we show that representative strains from all five C. difficile clades are able to produce p-cresol by two distinct mechanisms: (i) via fermentation of p-tyrosine and (ii) via uptake and turnover of exogenous p-HPA. We observed strain-specific differences in p-cresol production, resulting from differential efficiency of p-tyrosine fermentation; representatives of clade 3 (CD305) and clade 5 (M120) produced the highest levels of p-cresol via tyrosine metabolism, whereas the toxin A-/B+ isolate from clade 4 (M68) produced the lowest level of p-cresol. All five lineages share at least 97.3% homology across the hpdBCA operon, responsible for decarboxylation of p-HPA to p-cresol, suggesting that the limiting step in p-cresol production may result from tyrosine to p-HPA conversion. We identified that elevated intracellular p-HPA, modulated indirectly via CodY, controls p-cresol production via inducing the expression of HpdBCA decarboxylase ubiquitously in C. difficile populations. Efficient turnover of p-HPA is advantageous to C. difficile as p-HPA has a deleterious effect on the growth of C. difficile and other representative Gram-negative gut bacteria, transduced potentially by the disruption of membrane permeability and release of intracellular phosphate. This study provides insights into the importance of HpdBCA decarboxylase in C. difficile pathogenesis, both in terms of p-cresol production and detoxification of p-HPA, highlighting its importance to cell survival and as a highly specific therapeutic target for the inhibition of p-cresol production across C. difficile species.

Analysis of information asymmetry in the instructions for medical use on the example of Enterol

There may be variations in the texts of instructions for medical use of medicines (hereinafter – the instructions) containing the same active pharmaceutical ingredient, and registered in the pharmaceutical markets of different countries. First of all this applies to medicines manufactured in one country and distributed to other countries. Therefore, there are questions related to incorrect use of medicines due to the existing asymmetry of information. This problem is especially relevant in connection with tourist activity and labour migration. The aim of the study was to identify discrepancies in the instructions of Enterol, registered in different countries. The materials of the study were the registers of medicines in France, Ukraine, Belarus, Bulgaria, Poland, the Czech Republic and Russia, as well as the texts of instructions of Enterol (Biocodex, France), registered in the above countries. Methods of generalization, systematization, as well as contextual and comparative analysis were used. It was found that the original (French) instruction has two indications for the use of Enterol (treatment of acute diarrhea in children under 12 years old; prevention of antibiotic-associated diarrhea caused by Clostridium difficile or recurrence of diarrhea caused by Clostridium difficile), while Belarusian and Russian instructions have one indication. Polish instruction contains 5 indications, while Bulgarian and Czech – 8. The largest number of indications for the use of Enterol (11) is given in Ukrainian instruction. A contradictory information is in the sections «Therapeutic Indications», «Posology and method of administration», «Contraindications», «Undesirable effects», «Special warnings and precautions for use», including «Pregnancy and lactation». In the text of one instruction there is no information available in the text of another, and none of the instructions is identical to the instructions of Enterol, registered in France (country of manufacture). Actually, these information discrepancies, as well as the patient's perception of information, can lead to improper use of medicines and, as a consequence, drug-related problems. Thus, to minimize the asymmetry of information, the applicant during the registration of the drug must submit a notarized translation of the manufacturer's instructions.