• Journal of Microbiology and Biotechnology
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• v.32 no.1
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• pp.46-55
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• 2022

Clostridioides difficile infection (CDI) is a significant cause of hospital-acquired and antibiotic-mediated intestinal diseases and is a growing global public health concern. Overuse of antibiotics and their effect on normal intestinal flora has increased the incidence and severity of infections. Thus, the development of new, effective, and safe treatment options is a high priority. Here, we report a new probiotic strain, Bacillus amyloliquefaciens (BA PMC-80), and its in vitro/in vivo anti-C. difficile effect as a prospective novel candidate for replacing conventional antibiotics. BA PMC-80 showed a significant anti-C. difficile effect in coculture assay, and its cell-free supernatant (CFS) also exhibited a considerable anti-C. difficile effect with an 89.06 ㎍/ml 50% minimal inhibitory concentration (MIC) in broth microdilution assay. The CFS was stable and equally functional under different pHs, heat, and proteinase treatments. It also exhibited a high sensitivity against current antibiotics and no toxicity in subchronic toxicity testing in hamsters. Finally, BA PMC-80 showed a moderate effect in a hamster CDI model with reduced infection severity and delayed death. However, further studies are required to optimize the treatment condition of the hamster CDI model for better efficacy and identify the antimicrobial compound produced by BA PMC-80.

• Microbiology and Biotechnology Letters
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• v.47 no.4
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• pp.639-644
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• 2019

Diarrhea is one of the most common infectious diseases known worldwide. However, few studies have examined anaerobic diarrhea-causing bacteria (DB), which are difficult to culture. Recent advances in molecular biology have facilitated the detection and analysis of anaerobic DB. In this study, long-term trends in anaerobic DB were evaluated in Korea. From 2010 to 2018, symptoms of diarrhea reported were analyzed among patients hospitalized at the Dankook University Hospital in Korea. Results of multiplex polymerase chain reaction based on seasonality, age, overlapping infection, and other factors in patients were evaluated. DB were detected in 38.2% of 1716 stool specimens in the duration of the study. Of the pathogens detected using this method, 49.8% (n = 405/813) were anaerobic bacteria, including Clostridioides perfringens, Campylobacter spp., Clostridioides difficile toxin B, and Aeromonas spp. Among the four anaerobic bacteria, Clostridioides perfringens was the most commonly occurring (15.5%; n = 126/813). Detection rates of Clostridioides perfringens, Clostridioides difficile toxin B, and Aeromonas spp. were 34.1% (n = 22/55), 34.9% (n = 43/126), and 40.0% (n = 38/109), respectively. The detection rate of Campylobacter spp. (32.7%; n = 37/115) was the highest in patients between 10 and 20 years of age. The detection rate of anaerobic DB showed an increase in 2018 as compared with that in 2010, and the number of events of diarrhea caused by anaerobic DB also increased in this duration. Further studies are required to devise methods that might prevent the proliferation of anaerobic DB.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.25 no.5
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• pp.387-395
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• 2022

Purpose: Toxins produced by Clostridioides difficile infection (CDI) can cause enteritis and diarrhea. Although the number of pediatric CDI cases is increasing, the clinical management of pediatric CDI, including patient characteristics and prognosis, remains unclear. This study aimed to elucidate the background and clinical course of patients with CDI and evaluate the reliability of diagnostic tests in a tertiary pediatric hospital in Japan. Methods: We retrospectively analyzed the clinical data of children diagnosed with CDI between 2011 and 2021 at the Saitama Children's Medical Center in Saitama, Japan. Results: During the study period, 1,252 C. difficile antigen/toxin tests were performed, and 37 patients were diagnosed with CDI. The main underlying diseases among the patients were hematological and malignant disorders and gastrointestinal diseases, including inflammatory bowel disease (IBD) (59.4%). Two patients (5.4%) had an unremarkable medical history. Among the 37 patients, 27 (73.0%) were immunocompromised, 25 (67.6%) had a history of antibiotic use within the past two months, and 6 (16.2%) were negative on the initial test but were positive on the second test. Finally, 28 patients (75.7%) required primary antibiotic therapy only, and two patients with IBD required additional antibiotic therapy as secondary treatment. Conclusion: The number of pediatric patients with CDI is increasing. Both a comprehensive interview, including underlying diseases and history of antibiotic use, and an understanding of the features of clinical examinations should be emphasized to appropriately diagnose and treat CDI.

• Food Science of Animal Resources
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• v.40 no.4
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• pp.578-587
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• 2020

Clostridioides (Clostridium) difficile is a Gram (+), anaerobic, spore forming, rod shaped bacterium that can produce toxin. The objective of this study is to reveal the presence of C. difficile in meat products, to analyze the ribotype diversity by PCR and to evaluate the antibiotic susceptibility of isolated strains. The organism was isolated in 22 out of 319 (6.9%) examined meat product samples and 9 out of 22 (40.9%) isolates were identified as RT027 and all isolates had the ability of toxin production. In terms of antibiotic susceptibility, all isolates were susceptive to amoxicillin-clavulanic acid, tetracycline and vancomycin and 21 (95.4%) isolates to metronidazole. On the other hand, imipenem and cefotaxim resistance was observed in all. In conclusion, the results of this comprehensive study conducted in Turkey deduced the presence of C. difficile in different meat products. Therefore, these products can be evaluated as a potential contamination source of C. difficile from animals to humans especially for elders, youngsters, long terms wide spectrum antibiotic used and immuno-suppressed individuals.

• Biomedical Science Letters
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• v.26 no.3
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• pp.210-216
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• 2020

The purpose of this study was to evaluate GDH & Toxin (GDT) tests for the identification of the presence of Clostridioides difficile (C. difficile) as well as to detect whether any toxin was present in the feces of patients suspected of diarrhea associated with C. difficile. Data related to the results of toxin and culture (TC) tests and GDT tests conducted on patients with diarrhea and suspected CDI between January 2017 and august 2018, positive test rates, patient ages and sexes, whether the patients were hospitalized, and turnaround time (TAT) were analyzed retrospectively. Of the 7,554 total tests conducted for CDI diagnosis, 1,010 TC tests (14.9%) were positive, while 92 GDT tests (12.0%) were positive. Of these positive cases, 815 (80.7%) identified through TC test and 80 (87%) identified through GDT test were inpatients. also, among the patients with positive test results, 497 (49.2%) diagnosed through TC test and 45 (48.9%) diagnosed through GDT test were aged 61 years or older. The total time required to complete a TC test was 83.6 hours, while the time required for a GDT test was 11.2 hours, equating to an approximately three-day difference between the two tests. The detection of toxin-producing C. difficile is important in CDI diagnosis, but the commonly used Enzymeimmunoassay (EIA) toxin tests with low sensitivity result in delayed CDI diagnosis time. Therefore, primary screening tests for CDI diagnosis using the GDT method and secondary tests using additional methods are considered most effective.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.24 no.1
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• pp.81-89
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• 2021

Purpose: A steady increase in Clostridioides difficile enteritis (CDE) has been reported recently. CDE is associated with intestinal dysbiosis, and vitamin D receptors are known to play an important role in this microbial imbalance as immunological regulators. We investigated the difference in vitamin D levels between children with CDE and those with other acute infectious enteritis. Methods: This retrospective study was conducted on children below 18 years of age who visited the Gil hospital, underwent investigation to assess vitamin D levels, and had confirmed gastrointestinal infection between January 2015 and December 2018. Patients were divided into two groups: the "CDE group" (n=18) and the "other infectious enteritis group" (n=88); their clinical characteristics, other laboratory results, and vitamin D levels were analyzed. Results: There was no difference in gender, age, and seasonal distributions between the CDE and other infectious enteritis groups. Other laboratory results were not significantly different between two groups, excluding serum albumin level (4.52±0.45 g/dL vs. 4.31±0.28 g/dL, p=0.011). The mean 25-hydroxy vitamin D level in the CDE group was higher than that in the control group (18.75±8.11 ng/mL vs. 14.50±6.79 ng/mL, p=0.021). Conclusion: Vitamin D levels in the CDE group were lower than normal but higher than the other infectious enteritis group. These results suggested that CDE has a different mechanism or susceptibility associated with vitamin D in children, and even marginal changes in vitamin D levels can act as a risk factor for infection.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.25 no.1
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• pp.61-69
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• 2022

Purpose: Studies in adults have shown an increasing incidence of Clostridioides difficile infection (CDI) in patients hospitalized with acute pancreatitis (AP). There is lack of epidemiological data on CDI and its impact on hospitalized pediatric patients with AP. Methods: We analyzed the National Inpatient Sample and Kids' Inpatient Database between the years 2003 and 2016 and included all patients (age <21 years) with a primary diagnosis of AP using specific International Classification of Diseases codes. We compared clinical outcomes between children with CDI and those without CDI. Our primary outcome was severe AP and secondary outcomes included length of stay and hospital charges. Results: A total of 123,240 hospitalizations related to AP were analyzed and CDI was noted in 0.6% of the hospital. The prevalence rate of CDI doubled from 0.4% (2003) to 0.8% (2016), p=0.03. AP patients with CDI had increased comorbidities, and also underwent more invasive surgical procedures, p<0.05. AP patients with CDI had a higher in-hospital mortality rate and increased prevalence of severe AP, p<0.001. Multivariate regression models showed that CDI was associated with 2.4 times (confidence interval [CI]: 1.91 to 3.01, p<0.001) increased odds of severe AP. CDI patients had 7.24 (CI: 6.81 to 7.67, p<0.001) additional hospital days while incurring $59,032 (CI: 54,050 to 64,014, p<0.001) additional hospitalization charges. Conclusion: CDI in pediatric patients with AP is associated with adverse clinical outcomes and increased healthcare resource utilization. Further studies are needed to elucidate this association to prevent the development of CDI and to improve outcomes.

• Pediatric Infection and Vaccine
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• v.27 no.3
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• pp.158-170
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• 2020

Purpose: We evaluated the incidence and characteristics of Clostridioides difficile infection (CDI) in Korean children. Methods: Medical records of patients aged 2-18 years and diagnosed with CDI at a tertiary hospital between 2009 and 2018 were analyzed. The patients were classified into three CDI groups: community-acquired (CA), community onset-health care facility-associated (CO-HCFA), and healthcare facility onset (HO). Results: The incidence of CDI increased from 1.00 to 10.01 cases per 10,000 admissions from 2009 to 2018 (P<0.001). As compared to the CA group, the HO group had a higher frequency of operation and malignancy as predisposing factors (40.4% vs. 0.0%, P=0.001; and 27.7% vs. 0.0%, P=0.027, respectively), frequency and number of previous antibiotic use (97.9% vs. 31.3%, P<0.001; and 2 vs. 0, P<0.001, respectively), and median postdiagnosis hospital stay (13 vs. 5 days, P=0.008). The CO-HCFA group had a lower median age and higher frequency of malignancy than the CA group (5 vs. 13 years, P=0.012; and 30.8% vs. 0.0%, P=0.030, respectively). As compared to the HO group, the CA group had a higher frequency of abdominal pain and hematochezia (56.3% vs. 10.6%, P=0.001; and 50.0% vs. 10.6%, P=0.002, respectively), inflammatory bowel disease (68.8% vs. 2.1%, P=0.001), and intravenous metronidazole treatment (37.5% vs. 2.1%, P=0.001). Conclusions: With the increasing incidence of pediatric CDI, awareness regarding its epidemiology and clinical characteristics is important to manage nosocomial infections.

• Journal of Korean Medicine Rehabilitation
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• v.31 no.1
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• pp.47-57
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• 2021

Objectives This study was conducted to confirm the possibility of Clostridium difficile infection (CDI) treatment through natural herbal medicines. Methods After screening a total of 77 herbal medicines through the paper disc agar diffusion method, we selected the herbal medicines that showed a effectiveness compared to the positive control vancomycin. Afterwards, drugs that showed inhibitory effects compared to C. difficile without inhibition of Bifidobacterium bifidum and Lactobacillus plantarum, known as beneficial bacteria, were selected and minimal inhibitory concentration (MIC) was confirmed by applying the Broth microdilution method. Results The Coptidis Rhizoma, well known for its antimicrobial effect, was found to have antimicrobial effects on C. difficile, but also had inhibitory effects on the beneficial bacterium B. bifidum. 30% ethanol extraction Crataegi fructus, Corni fructus and Mume fructus had antimicrobial effects on C. difficile without inhibiting the beneficial bacteria B. bifidum and L. plantarum. The MIC values of 30% ethanol extraction Crataegi fructus, Corni fructus and Mume fructus were found to be 10 mg/mL, 20 mg/mL and 5 mg/mL, respectively. Conclusions Crataegi fructus, Corni fructus and Mume fructus were identified as candidate medicines for C. difficile. Further researchs will need to be done in vivo, and to find an optimal extraction method accompanied by economic evaluation.