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  1. Detection of Clostridium difficile from an enhancement broth by gas-liquid chromatography
    Hassan H, Cheng SC
    Malays J Pathol, 1991 Jun;13(1):29-31.
    PMID: 1795558
    Gas-Liquid chromatography (GLC) was used to detect the presence of isocaproic acid produced by Clostridium difficile from 54 stool samples grown in cycloserine-cefoxitin broth. Isocaproic acid was detected in 12 samples of which 5 were confirmed to be Clostridium difficile by culture and biochemical tests. The detection of isocaproic acid by GLC together with the presence of presumptive colonies on primary selective culture media provides a more rapid laboratory diagnosis for Clostridium difficile.
    Matched MeSH terms: Clostridium difficile/isolation & purification*; Clostridium difficile/metabolism
  2. Fulltext Diversity, Dynamics and Therapeutic Application of Clostridioides difficile Bacteriophages
    Nale JY, Thanki AM, Rashid SJ, Shan J, Vinner GK, Dowah ASA, et al.
    Viruses, 2022 Dec 12;14(12).
    PMID: 36560776 DOI: 10.3390/v14122772
    Clostridioides difficile causes antibiotic-induced diarrhoea and pseudomembranous colitis in humans and animals. Current conventional treatment relies solely on antibiotics, but C. difficile infection (CDI) cases remain persistently high with concomitant increased recurrence often due to the emergence of antibiotic-resistant strains. Antibiotics used in treatment also induce gut microbial imbalance; therefore, novel therapeutics with improved target specificity are being investigated. Bacteriophages (phages) kill bacteria with precision, hence are alternative therapeutics for the targeted eradication of the pathogen. Here, we review current progress in C. difficile phage research. We discuss tested strategies of isolating C. difficile phages directly, and via enrichment methods from various sample types and through antibiotic induction to mediate prophage release. We also summarise phenotypic phage data that reveal their morphological, genetic diversity, and various ways they impact their host physiology and pathogenicity during infection and lysogeny. Furthermore, we describe the therapeutic development of phages through efficacy testing in different in vitro, ex vivo and in vivo infection models. We also discuss genetic modification of phages to prevent horizontal gene transfer and improve lysis efficacy and formulation to enhance stability and delivery of the phages. The goal of this review is to provide a more in-depth understanding of C. difficile phages and theoretical and practical knowledge on pre-clinical, therapeutic evaluation of the safety and effectiveness of phage therapy for CDI.
    Matched MeSH terms: Clostridium difficile*
  3. A case report of improvement on ADHD symptoms after fecal microbiota transplantation with gut microbiome profiling pre- and post-procedure
    Hooi SL, Dwiyanto J, Rasiti H, Toh KY, Wong RKM, Lee JWJ
    Curr Med Res Opin, 2022 Nov;38(11):1977-1982.
    PMID: 36164761 DOI: 10.1080/03007995.2022.2129232
    BACKGROUND: Recent studies demonstrate the association of the gut microbiome in regulating interactions between the central nervous system and intestinal function. Individuals with attention-deficit hyperactivity disorder (ADHD) have been shown to have unique gut microbial signature, with depletion of beneficial commensal microbes. Fecal microbiota transplant (FMT) restores the imbalanced gut microbiome and may replete missing microbes to increase production of hormones and neurotransmitters regulating human behavior and cognition.

    RESEARCH DESIGN & METHODS: Here, we present an interesting case of a 22-year-old woman treated with FMT primarily to treat recurrent Clostridioides difficile infection, which coincidentally alleviated her ADHD symptoms. We also present the pre- and post-FMT gut microbiota profiles conducted using shotgun metagenomic sequencing on the patient's fecal samples to thereby highlight potential microbial-associated mechanisms associated with the relief of ADHD symptoms.

    RESULTS & CONCLUSIONS: Our case report provides preliminary evidence regarding the use of FMT in a patient with C. difficile and ADHD. We speculate that gut microbiome modulation, in particular the gain or loss of specific microbial species and pathways involving the metabolism of SCFAs, tryptophan and GABA, may merit further exploration as a potential therapeutic strategy for ADHD.

    Matched MeSH terms: Clostridium difficile*
  4. Molecular epidemiology of Clostridium difficile isolated from piglets
    Putsathit P, Neela VK, Joseph NMS, Ooi PT, Ngamwongsatit B, Knight DR, et al.
    Vet Microbiol, 2019 Oct;237:108408.
    PMID: 31585650 DOI: 10.1016/j.vetmic.2019.108408
    Information on the epidemiology of C. difficile infection (CDI) in South-East Asian countries is limited, as is data on possible animal reservoirs of C. difficile in the region. We investigated the prevalence and molecular epidemiology of C. difficile in piglets and the piggery environment in Thailand and Malaysia. Piglet rectal swabs (n = 224) and piggery environmental specimens (n = 23) were collected between 2015 and 2016 from 11 farms located in Thailand and Malaysia. All specimens were tested for the presence of C. difficile with toxigenic culture. PCR assays were performed on isolates to determine the ribotype (RT), and the presence of toxin genes. Whole genome sequencing was used on a subset of isolates to determine the evolutionary relatedness of RT038 (the most prevalent RT identified) common to pigs and humans from Thailand and Indonesia. C. difficile was recovered from 35% (58/165) and 92% (54/59) of the piglets, and 89% (8/9) and 93% (13/14) of the environmental specimens from Thailand and Malaysia, respectively. All strains from Thailand, and 30 strains from Malaysia (23 piglet and 7 environmental isolates) were non-toxigenic. To our knowledge, this is the first and only report with a complete lack of toxigenic C. difficile among piglets, a feature which could have a protective effect on the host. The most common strain belonged to RT038 (ST48), accounting for 88% (51/58) of piglet and 78% (7/9) of environmental isolates from Thailand, and all 30 isolates tested from Malaysia. Piglet RT038 isolates from Thailand and Malaysia differed by only 18 core-genome single nucleotide variants (cgSNVs) and both were, on average, 30 cgSNVs different from the human strains from Thailand and Indonesia, indicating a common ancestor in the last two decades.
    Matched MeSH terms: Clostridium difficile/genetics*
  5. Clostridium difficile-related necrotizing pseudomembranous enteritis in association with Henoch-Schonlein purpura
    Boey CC, Ramanujam TM, Looi LM
    J Pediatr Gastroenterol Nutr, 1997 Apr;24(4):426-9.
    PMID: 9144126
    Matched MeSH terms: Clostridium difficile/isolation & purification
  6. Biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxy-4-methylvalerate) by recombinant Escherichia coli expressing leucine metabolism-related enzymes derived from Clostridium difficile
    Saika A, Watanabe Y, Sudesh K, Tsuge T
    J Biosci Bioeng, 2014 Jun;117(6):670-5.
    PMID: 24484910 DOI: 10.1016/j.jbiosc.2013.12.006
    An obligate anaerobic bacterium Clostridium difficile has a unique metabolic pathway to convert leucine to 4-methylvalerate, in which 4-methyl-2-pentenoyl-CoA (4M2PE-CoA) is an intermediate of this pathway. 4M2PE-CoA is also able to be converted to 3-hydroxy-4-methylvalerate (3H4MV), a branched side chain monomer unit, for synthesis of polyhydroxyalkanoate (PHA) copolymer. In this study, to synthesize 3H4MV-containing PHA copolymer from leucine, the leucine metabolism-related enzymes (LdhA and HadAIBC) derived from C. difficile and PHA biosynthesis enzymes (PhaPCJAc and PhaABRe) derived from Aeromonas caviae and Ralstonia eutropha were co-expressed in the codon usage-improved Escherichia coli. Under microaerobic culture conditions, this E. coli was able to synthesize P(3HB-co-12.2 mol% 3H4MV) from glucose with the supplementation of 1 g/L leucine. This strain also produced P(3HB-co-12.6 mol% 3H4MV) using the culture supernatant of leucine overproducer E. coli strain NS1391 as the medium for PHA production, achieving 3H4MV copolymer synthesis only from glucose. Furthermore, we tested the feasibility of the 3H4MV copolymer synthesis in E. coli strain NS1391 from glucose. The recombinant E. coli NS1391 was able to synthesize P(3HB-co-3.0 mol% 3H4MV) from glucose without any leucine supplementation. This study demonstrates the potential of the new metabolic pathway for 3H4MV synthesis using leucine metabolism-related enzymes from C. difficile.
    Matched MeSH terms: Clostridium difficile/enzymology; Clostridium difficile/genetics*
  7. Fulltext Indication of acid suppression therapy and predictors for the prophylactic use of protonpump inhibitors vs. histamine-2 receptor antagonists in a Malaysian tertiary hospital
    Oh AL, Tan AG, Phan HS, Lee BC, Jumaat N, Chew SP, et al.
    Pharm Pract (Granada), 2015 Apr-Jun;13(3):633.
    PMID: 26445624 DOI: 10.18549/PharmPract.2015.03.633
    Proton-pump inhibitors (PPI) and histamine-2 receptor antagonists (H2RA) are common acid suppressants used in gastrointestinal disorders. The trend of usage in Malaysia has changed from predominantly H2RA to PPI from 2007 to 2008, 3.46 versus 2.87 and 2.99 versus 3.24 DDD (Defined Daily Dose)/1000 population/day respectively. This raises concerns as PPI overutilization amounts to higher cost expenditure and are associated with various untoward consequences such as Clostridium difficile-associated diarrhea, pneumonia, and osteoporosis.
    Matched MeSH terms: Clostridium difficile
  8. Fulltext A review on the effects of probiotics and antibiotics towards Clostridium difficile infections
    Hazirah, A., Loong, Y.Y., Rushdan, A.A., Rukman, A.H., Yazid, M.M.
    Pertanika Journal of Science & Technology, 2013;21(2):293-302.
    MyJurnal
    Clostridium difficile can cause severe diseases with significant morbidity and mortality in infected patients.
    The rate of Clostridium difficile infection is high in North America and European countries. Metronidazole
    and vancomycin have been recommended as the treatments of choice since 1990s. Recurrent infection
    due to Clostridium difficile is common after several days of antibiotic administration. Probiotics have
    been used in these patients as an adjunct treatment with some successful findings. However, a detailed
    investigation on the use of probiotic for infected patients is still needed, particularly for its real efficacy.
    Matched MeSH terms: Clostridium difficile
  9. Fulltext High Prevalence of Toxigenic and Nontoxigenic Clostridium difficile Strains in Malaysia
    Riley TV, Collins DA, Karunakaran R, Kahar MA, Adnan A, Hassan SA, et al.
    J Clin Microbiol, 2018 Jun;56(6).
    PMID: 29563206 DOI: 10.1128/JCM.00170-18
    Accumulating evidence shows a high prevalence of Clostridium difficile in Southeast Asia associated with a range of clinical presentations. However, severe infections are rarely reported. We investigated C. difficile infection (CDI) across four hospitals in Kuala Lumpur and Kota Bharu, Malaysia. Enzyme immunoassays for glutamate dehydrogenase (GDH) and toxin A or B were performed on diarrheal stool specimens collected from patients in 2015 and 2016. Specimens were also cultured and isolates of C. difficile characterized by PCR ribotyping and detection of toxin genes. In total, 437 specimens were collected and fecal toxin was detected in 3.0%. A further 16.2% of specimens were GDH positive and toxin negative. After culture, toxigenic strains were isolated from 10.3% and nontoxigenic strains from 12.4% of specimens. The most prevalent PCR ribotypes (RTs) were RT 017 (20.0%) and RT 043 (10.0%). The high prevalence of RT 017 and nontoxigenic strains in Malaysia and in neighboring Thailand and Indonesia suggests that they localize to the region of Southeast Asia, with an implication that they may mediate the burden of CDI in the region.
    Matched MeSH terms: Clostridium difficile/genetics*; Clostridium difficile/isolation & purification*
  10. Additional oligofructose/inulin does not increase faecal bifidobacteria in critically ill patients receiving enteral nutrition: a randomised controlled trial
    Majid HA, Cole J, Emery PW, Whelan K
    Clin Nutr, 2014 Dec;33(6):966-72.
    PMID: 24290345 DOI: 10.1016/j.clnu.2013.11.008
    Patients with diarrhoea during enteral nutrition (EN) have been shown to have low faecal bifidobacteria concentrations. Oligofructose/inulin selectively stimulate the growth of bifidobacteria in healthy humans. This study investigates the effect of additional oligofructose/inulin on the gastrointestinal microbiota, short-chain fatty acids (SCFA) and faecal output in patients receiving EN.
    Matched MeSH terms: Clostridium difficile/isolation & purification
  11. Fulltext 2019 update of the WSES guidelines for management of Clostridioides (Clostridium) difficile infection in surgical patients
    Sartelli M, Di Bella S, McFarland LV, Khanna S, Furuya-Kanamori L, Abuzeid N, et al.
    World J Emerg Surg, 2019 02 28;14:8.
    PMID: 30858872 DOI: 10.1186/s13017-019-0228-3
    In the last three decades, Clostridium difficile infection (CDI) has increased in incidence and severity in many countries worldwide. The increase in CDI incidence has been particularly apparent among surgical patients. Therefore, prevention of CDI and optimization of management in the surgical patient are paramount. An international multidisciplinary panel of experts from the World Society of Emergency Surgery (WSES) updated its guidelines for management of CDI in surgical patients according to the most recent available literature. The update includes recent changes introduced in the management of this infection.
    Matched MeSH terms: Clostridium difficile/pathogenicity*
  12. Prevalence of Clostridium difficile infection and colonization in a tertiary hospital and elderly community of North-Eastern Peninsular Malaysia
    Zainul NH, Ma ZF, Besari A, Siti Asma H, Rahman RA, Collins DA, et al.
    Epidemiol Infect, 2017 Oct;145(14):3012-3019.
    PMID: 28891459 DOI: 10.1017/S0950268817002011
    Little is known about Clostridium difficile infection (CDI) in Asia. The aims of our study were to explore (i) the prevalence, risk factors and molecular epidemiology of CDI and colonization in a tertiary academic hospital in North-Eastern Peninsular Malaysia; (ii) the rate of carriage of C. difficile among the elderly in the region; (iii) the awareness level of this infection among the hospital staffs and students. For stool samples collected from hospital inpatients with diarrhea (n = 76) and healthy community members (n = 138), C. difficile antigen and toxins were tested by enzyme immunoassay. Stool samples were subsequently analyzed by culture and molecular detection of toxin genes, and PCR ribotyping of isolates. To examine awareness among hospital staff and students, participants were asked to complete a self-administered questionnaire. For the hospital and community studies, the prevalence of non-toxigenic C. difficile colonization was 16% and 2%, respectively. The prevalence of CDI among hospital inpatients with diarrhea was 13%. Out of 22 C. difficile strains from hospital inpatients, the toxigenic ribotypes 043 and 017 were most common (both 14%). In univariate analysis, C. difficile colonization in hospital inpatients was significantly associated with greater duration of hospitalization and use of penicillin (both P < 0·05). Absence of these factors was a possible reason for low colonization in the community. Only 3% of 154 respondents answered all questions correctly in the awareness survey. C. difficile colonization is prevalent in a Malaysian hospital setting but not in the elderly community with little or no contact with hospitals. Awareness of CDI is alarmingly poor.
    Matched MeSH terms: Clostridium difficile/isolation & purification*
  13. Radical hysterectomy and pelvic lymphadenectomy for early invasive cancer of the cervix - 14-year experience
    Sivanesaratnam V, Sen DK, Jayalakshmi P, Ong G
    Int. J. Gynecol. Cancer, 1993 Jul;3(4):231-238.
    PMID: 11578351
    During a 14-year period, 397 radical hysterectomies and pelvic lymphadenectomies were performed for early invasive carcinoma of the cervix. Twenty-one patients were in stage IA2 with lymphatic/vascular channel permeation (5.2%), 340 in stage IB (85.6%) and 34 in early stage 2A disease (8.5%). Eighteen patients (4.5%) were pregnant. Adenocarcinoma comprised 26.9% of cases. The mean operative time was 4.14 h; the intraoperative blood loss was less than 1.51 in 77.3% patients. There was no operative mortality; one patient died 3 weeks after surgery from clostridium difficile enterocilitis. Eleven patients (2.7%) developed venous thrombosis; severe lymphedema occurred in four (1%). The incidence of uretero-vaginal fistula was 0.2% and that of vesico-vaginal fistula 0.5%. Ovarian metastases were noted in 4.3% of cases with adenocarcinoma. Sixty-six patients had positive nodes (16.6%). Five-year survival in patients with more than 2 positive nodes was 68%. The use of adjuvant chemotherapy in patients with 'high risk' factors resulted in survival rates approaching those without risk factors. Neo-adjuvant chemotherapy was used in 10 patients with large bulky tumors; the results were favorable. Recurrences occurred in 47 patients (11.8%); 36 patients have died (9.1%). Age did not appear to influence survival. The overall 5-year survival was 92.2%.
    Matched MeSH terms: Clostridium difficile
  14. Fulltext Recommendations for the adjuvant use of the poly-antibiotic-resistant probiotic Bacillus clausii (O/C, SIN, N/R, T) in acute, chronic, and antibiotic-associated diarrhea in children: consensus from Asian experts
    De Castro JA, Kesavelu D, Lahiri KR, Chaijitraruch N, Chongsrisawat V, Jog PP, et al.
    Trop Dis Travel Med Vaccines, 2020;6:21.
    PMID: 33110611 DOI: 10.1186/s40794-020-00120-4
    This paper proposes recommendations for probiotics in pediatric gastrointestinal diseases in the Asia-Pacific region. Evidence-based recommendations and randomized controlled trials in the region are included. Cultural aspects, health management issues and economic factors were also considered. Final recommendations were approved by utilizing a modified Delphi process and applying the Likert scale in an electronic voting process. Bacillus clausii was recommended as an adjunct treatment with oral rehydration solution for acute viral diarrhea. B. clausii may also be considered for prevention of antibiotic-associated diarrhea, Clostridium difficile-induced diarrhea, and as adjunct treatment of Helicobacter pylori. There is insufficient evidence for recommendations in other conditions. Despite a diversity of epidemiological, socioeconomical and health system conditions, similar recommendations currently apply to most Asia-Pacific countries. Ideally, these need to be validated with local randomized-controlled trials.
    Matched MeSH terms: Clostridium difficile
  15. Fulltext Clostridium difficile Infection: clinico-epidemiological perspective
    Syuhada, N., Azimatun, N.A., Alfizah, H., Tzar, M.N., Ramliza, R.
    Medicine & Health, 2013;8(2):64-72.
    MyJurnal
    Clostridium difficile infection (CDI) causes mild to severe diarrhoea and pseudomembranous colitis in patients who had prior antibiotic exposure. Despite CDI being prevalent worldwide, its epidemiological data is scanty in Malaysia. This study aimed to determine the prevalence and incidence of CDI at Universiti Kebangsaan Malaysia Medical Centre (UKMMC). Stool specimens from 147-suspected CDI patients were obtained from 1 November 2011 until 31 October 2012. The presence of C. difficile toxin A and/or B were detected using a commercial immunochromatographic kit (Wampole™ Tox A/B Quik Chek). Surveillance data was collected from patients’ medical records to establish the demographic and clinical characteristics. The overall prevalence and incidence of CDI in UKMMC was 6.1% and 5.2 cases per 10 000 patient-days, respectively. Among nine CDI patients, 77.8% were males and 55.6% were Chinese. CDI was most common in medical wards (88.9%). The median age was 60 years and the median length of hospital stay was 13 days. Majority (88.9%) of CDI patients received antibiotics eight weeks prior to CDI. Penicillin-beta-lactamase inhibitors were the most common antecedent antibiotics. Five (55.6%) CDI patients received acid suppressant medications. The in-hospital mortality rate was 22.2%. In conclusion, the prevalence and incidence of CDI at UKMMC is relatively low and occurs sporadically.
    Matched MeSH terms: Clostridium difficile
  16. Fulltext A standardised model for stool banking for faecal microbiota transplantation: a consensus report from a multidisciplinary UEG working group
    Keller JJ, Ooijevaar RE, Hvas CL, Terveer EM, Lieberknecht SC, Högenauer C, et al.
    United European Gastroenterol J, 2021 Mar;9(2):229-247.
    PMID: 33151137 DOI: 10.1177/2050640620967898
    BACKGROUND: Faecal microbiota transplantation is an emerging therapeutic option, particularly for the treatment of recurrent Clostridioides difficile infection. Stool banks that organise recruitment and screening of faeces donors are being embedded within the regulatory frameworks described in the European Union Tissue and Cells Directive and the technical guide to the quality and safety of tissue and cells for human application, published by the European Council.

    OBJECTIVE: Several European and international consensus statements concerning faecal microbiota transplantation have been issued. While these documents provide overall guidance, we aim to provide a detailed description of all processes that relate to the collection, handling and clinical application of human donor stool in this document.

    METHODS: Collaborative subgroups of experts on stool banking drafted concepts for all domains pertaining to stool banking. During a working group meeting in the United European Gastroenterology Week 2019 in Barcelona, these concepts were discussed and finalised to be included in our overall guidance document about faecal microbiota transplantation.

    RESULTS: A guidance document for all domains pertaining to stool banking was created. This document includes standard operating manuals for several processes involved with stool banking, such as handling of donor material, storage and donor screening.

    CONCLUSION: The implementation of faecal microbiota transplantation by stool banks in concordance with our guidance document will enable quality assurance and guarantee the availability of donor faeces preparations for patients.

    Matched MeSH terms: Clostridium difficile
  17. Hospital-acquired Clostridium difficile infection among patients with type 2 diabetes mellitus in acute medical wards
    Hassan SA, Rahman RA, Huda N, Wan Bebakar WM, Lee YY
    J R Coll Physicians Edinb, 2013;43(2):103-7.
    PMID: 23734349 DOI: 10.4997/JRCPE.2013.203
    Clostridum difficile (C. difficile) infection is increasingly seen among hospitalised patients with type 2 diabetes mellitus but its rate and associated risk factors are not known. We aimed to determine the rate and characteristics of hospital-acquired C. difficile infection in subjects with type 2 diabetes mellitus admitted into acute medical wards.
    Matched MeSH terms: Clostridium difficile*
  18. Fulltext Prevalence of Clostridium difficile toxin in diarhoeal stool samples of patients from a tertiary hospital in North Eastern Penisular Malaysia
    Hassan SA, Othman N, Idris FM, Abdul Rahman Z, Maning N, Abdul Rahman R, et al.
    Med J Malaysia, 2012 Aug;67(4):402-5.
    PMID: 23082450 MyJurnal
    This study describes the prevalence of Clostridium difficile toxin (CDT) in loose stool samples from inpatients aged more than two years of a tertiary hospital. A total of 175 samples that had been examined were from stool samples that were sent to the Medical Microbiology & Parasitology Laboratory for various clinical indications. The toxin was detected by a commercial immunochromatograhic test, and the patients' demography, clinical features, treatment and outcomes were analyzed from their medical records. Clostridium difficile toxin was positive in 24 (13.7%) of the stool samples. Male and female were 11 (45.8%) and 13 (54.2%) respectively, with the majority of them aged more than 50 years. Most were from medical wards (n = 21, 87.5%), with the rest from surgical wards (n = 2, 8.3%) and intensive care units (n = 1, 3.4%). All the CDT positive patients had history of prior antibiotic usage within 6 weeks before the detection of the toxin. The mean duration of antibiotics usage was 17.75 (+/- 13.75) days, while the mean duration of diarrhea was 5.21((+/- 5.85) days. Eighteen patients had underlying medical illnesses that were diabetes mellitus, chronic renal disease, hypertension, ischaemic heart disease, cerebrovascular disease and malignancy; with seven of them being CDT positive while on chemotherapy. Stool occult blood test was positive in 15 patients whereas presence of pus cells in the CD positive stool samples were detected in 21 patients. The duration of hospitalization among the patients was 27.96 (+/- 23.22) days.
    Matched MeSH terms: Clostridium difficile
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