Clostridium Research Articles

Establishment of digital polymerase chain reaction-ribotyping and database for Clostridium difficile genotyping

Objective To develop a digital polymerase chain reaction (PCR)-ribotyping method and database for Clostridium difficile genotyping. Methods Sequencer based fluorescence capillary gel electrophoresis was used, instead of agarose gel electrophoresis, to establish the digital PCR-ribotyping of Clostridium difficile. Forty Clostridium difficile reference strains, consisting of 10 PCR-ribotypes (RT), were genotyped by the new digital PCR-ribotyping method to set-up the database. Results The sequencer based fluorescence capillary gel electrophoresis correctly detected PCR-ribotyping products of the 40 reference strains, and showed as digital figure; significant differences of these digital figures were found between the 10 RT. High similar digital figures were shown in twenty-one RT027 strains, three RT002 strains and two RT014 strains. However, seven RT001 strains were typed as four subtypes, and two RT014 strains as two subtypes, respectively. Conclusion A digital PCR-ribotyping, and a reference database consisting of 10 RT are successfully established. Key words: Clostridium difficile; PCR-ribotyping; Fluorescence capillary gel electrophoresis; Digital; Database

Frozen Feces Are Effective for Recurrent C. Difficile Infection

Fecal microbiota transplantation (FMT) is an effective treatment for recurrent Clostridium difficile infection. Obtaining fresh stool from volunteer donors is expensive and inconvenient because it requires screening donors for a number of infections. In the current randomized, controlled noninferiority trial, investigators assessed clinical resolution in 219 patients with recurrent or refractory C. difficile …

Hygienic characteristics and microbiological hazard identification in horse and donkey raw milk.

Today the interest toward horse (Equus caballus) and donkey (Equus asinus) milk for human consumption is receiving a renewed attention because of its particular composition, hypoallergenicity, and nutraceutical properties. The realistic perspective of global use of this aliment in balanced diets, especially for infancy and geriatrics, poses the need for a more in depth knowledge on milk hygiene and on the health status of dairy animals, as a prerequisite of consumers' safety. The aim of this paper was to review the available literature on the health and hygiene parameters as well as on the potential microbiological hazards in horse and donkey milk and the risks related to their consumption. Both microbial contamination and somatic cell count are reasonably low in equine milk and also the presence of pathogens, like Escherichia coli O157, Salmonella spp., Campylobacter spp., Yersinia enterocolitica, Brucella spp., Mycobacterium spp., Bacillus cereus, Cronobacter sakazakii, Streptococcus equi subsp. zooepidemicus, Rhodococcus equi, Streptococcus dysgalactiae subsp. equisimilis, Clostridium difficile and Burkholderia mallei is low. However, in those regions of the world where the prevalence of Brucella spp. and Rhodococcus equi is high, the alimentary risks could increase. Similarly, in areas with higher incidence of immunocompromised people, the increased risks should be warned not only for pathogens but also for opportunistic microbiota.

Efficacy studies on peracetic acid against pathogenic microorganisms

Background: The peracetic acid (PAA) has antimicrobial activity against bacteria and fungi including spores and is envirosafe. Despite its widespread use in food industry and effluent treatment, it is not widely used in hospitals. The present work was aimed to find out its efficacy against hospital pathogens, bacterial and fungal spores and mycobacteria. Methods: Multidrug-resistant, wild hospital isolates of Gram-positive, Gram-negative bacteria, tough organisms such as Mycobacterium fortuitum, Mycobacterium tuberculosis and Candida albicans and spores of Bacillus subtilis, Clostridium perfringens and Aspergillus niger were checked by modified Kelsey-Sykes suspension test. For vegetative bacteria, exposure time was 1 min. For bacterial and fungal spores, Candida and Mycobacteria, exposure time varied from 10 to 30 min. Results: More than 5-log reduction was seen for vegetative bacteria just after 1 min exposure to PAA. Ten minute exposure to PAA could inactivate 99.5% bacterial and fungal spores. Mycobacteria were inactivated within 10 min of exposure to PAA. PAA rapidly inactivates pathogenic bacteria within 1 min and inactivates mycobacteria and fungi within 10 min and sterilises spores within 30 min and remains active in the presence of proteins. Conclusions: It is economic, eco-friendly and deserves major share in hospital disinfection.

Toxinotype of Clostridium difficile in Taiwan

Clostridium difficile is a pathogen highly associated with nosocomial infection; it causes diseases ranging from self-limited diarrhea to pseudomembranous colitis. Recently, the hyper-virulent strain of C. difficile has appeared in Taiwan. In addition, a C. difficile nosocomial outbreak was reported at one southern hospital. In this study, 59 strains of toxigenic C. difficile with known PCR-ribotypes were collected during 2008-2009 from eight and two hospitals from central and southern Taiwan, and characterized by toxinotyping and Clostridium difficile binary toxin (CDT) detection. Of 59 strains, 48 strains produced A+B+ toxin, and 11 produced A-B+ toxin. Among A+B+ toxin producing strains, 46 strains were toxinotype 0, 1 strain was toxinotype VI, and 1 strain was new toxinotype (toxinotype TN1). Among A-B+ toxin producing strains, 6 strains were toxinotype VIII, 3 strains were toxinotype V-like, and 2 strains belonged to a new type (toxinotype TN2). The strains of toxinotype 0 belonged to several PCR-riboytpes, whereas the strains of toxinotype VIII belonged to one PCR-ribotype, this indicated toxinotype VIII strains belonged to the same clonal origin. Six (10.2%) of 59 strains had CDT gene; among them, 2 strains were A+B+ toxin producing, 4 strains were A-B+ toxin producing. This study found that prevalence of A-B+ strains in Taiwan was similar to those in other Asian countries, but higher than those in Europe and USA. The strains of toxinotype V-like and VI in Taiwan was not reported in other Asian countries, neither the 2 new toxinotypes TN1 and TN2.

The burden of Clostridium difficile infection in Thais: nationwide survey

Background: Data on the incidence and burden of Clostridium difficile infection (CDI) in Asia is limited. Objectives: To evaluate the incidence and burden of CDI in Thailand. Methods: We used 2010 Nationwide Hospital Admission Data, which included the diagnosis of digestive disorders from various causes coded using the ICD-10. Patients with a diagnosis of Clostridium difficile (ICD10-A07) aged >18 years, were included. Their baseline characteristics, clinical outcomes, and risk factors for CDI were analyzed. Length of hospital stay (LOS), mortality rate, and hospital expenses were used as indicators to evaluate the burden of CDI in Thailand. Results: Of 4,863,935 admissions in 2010, 554 patients in 570 admissions (0.01%) were diagnosed with CDI. Of these, 106 (19.1%) died during the index hospitalization, and 98.1% had at least one comorbidity. The mean LOS for patients with CDI was longer than with other colitis (P who died during the index admission, compared with those who survived during the index admission (P = 0.04). The hospital expense for those who died was significantly higher than for those who survived (P From a multivariate analysis, age ?85 years old, comorbidity, and sepsis were risk factors for mortality during admission with adjusted odds ratios of 2.40, 7.4, and 5.14, respectively. Conclusions: The calculated burden of CDI in Thais is high; although the incidence of CDI is lower in Thailand than in Western countries. The mortality relates to the elderly age-group and comorbidity, especially sepsis. Keywords: Burden, Clostridium difficile infection, nationwide study, Southeast Asia, Thailand, epidemiology

PPI Use and Changes in Bacterial Flora Associated with C. difficile Infection

Proton-pump inhibitor (PPI) use is believed to increase the risk for Clostridium difficile infection (CDI), presumably by altering the bacteria normally found in the gut. To further investigate this presumption, researchers conducted a prospective, open-label trial involving 12 healthy volunteers. Stool samples were obtained at baseline and 4 weeks, after which all participants …

Power generation from pre-treated empty fruit bunch using single chamber microbial fuel cell

The utilization of lignocellulosic biomass (LB) as the substrate in Microbial Fuel Cell (MFC) have been receive growing interest due to the vast amount of abundant LB side-product that are generated yearly from agriculture field. In this paper the author will be discussing the utilization of Empty Fruit Bunches (EFB), which is one of the LB waste generated from palm oil industry in single chamber MFC (SCMFC) to generate bioelectricity. The microbes used in this study were equivalent volume mixtures of Gram-positive Bacillus E1 and Clostridium cellulolyticum (CC). To enhance the efficiency of the microbes to utilize the EFB for bioelectricity generation, the EFB used were pre-treated first with several pre-treatment method which include physical pre-treatment, hot water pre-treatment and alkaline pre-treatment. Different amount of EFB were tested which include 1.5 g, 3.5 g and 5.5 g of EFB under similar condition with total working volume of 250 mL for anode chamber. Resistor of range 100-100,000 O was connected to the MFC to calculate the current and power generated from the system. Results indicate maximum power was achieved at the value of up to 0.7 W/m2 at above 1.0 A/m2.for all pre-treated EFB tested. The highest power was achieved at 100 O using lower concentration of EFB with a value of 0.678 W/m2. In conclusion, EFB is a feasible substrate for MFC and more studies on improvement of the power generation in progress for larger scale application.

Therapeutic effects and the possible mechanism of fecal transplantation on rats with Clostridium difficile-associated pseudomembranous colitis

Objective To investigate the therapeutic effects and the possible mechanism of fecal transplantation on rats with Clostridium difficile-associated pseudomembranous colitis. Methods A total of 40 Sprague-Dawley rats were divided into four groups including the healthy control group, model group, fecal transplant treatment group and vancomycin treatment group. Rats in three experimental groups were subcutaneously injected with clindamycin phosphate (10 mg), followed by treatment with toxin producing Clostridium difficile (ACTT43255) enema 24 hours later. The rats in fecal transplant treatment group and vancomycin treatment group were respectively treated with fecal suspension and vancomycin one day after modeling. The rats were fasted for one day after the last administration and then executed. The levels of potassium ion (K), sodium ion (Na), albumin (ALB), white blood cells (WBC), C-reaction protein (CRP), interleukin-1β (IL-1β), interleukin-10 (IL-10), interleukin-12 (IL-12) and interleukin-17 (IL-17) as well as the percentage of neutrophils (N%) in serum samples were detected. The colon tissue samples were collected for pathology examination. Results The rat model of pseudomembranous colitis was successfully established by subcutaneous injection of clindamycin in combination with toxin-producing Clostridium difficile (ACTT43255) enema. The signs of intestinal inflammation including serious weight loss, remarkably shortened colon length and significantly increased colon wet weight index were observed in rats from the model group (P<0.05). Compared with the rats from model group, the rats received fecal transplant showed significantly increased levels of K, ALB, IL-10 and IL-10/IL-12 in serum and decreased levels of WBC, N%, CRP, IL-1β and IL-17 (P<0.05). Conclusion Fecal transplantation was proved to be an effective approach for the treatment of pseudomembranous colitis. The therapeutic mechanism might due to its impacts on serum inflammatory factors. Key words: Fecal transplantation; Pseudomembranous colitis; Inflammatory factor

Reducing Antibiotic-Resistant Infections: All Together Now!

We are rapidly approaching an era when we will run out of weapons against antibiotic-resistant bacteria and Clostridium difficile.

AB009. Xiaflex as a treatment for Peyronie’s disease

Peyronie disease can be physically and psychologically devastating for subjects and their partners. Collagenase Clostridium histolyticum (CCh), a purified mixture of AUX-I and II collagenases, is an intralesional, minimally invasive intervention with evidence of tolerability and efficacy in subjects with PD and is approved by the United States Food and Drug Administration for use in adults who have Peyronie Disease and Dupuytren contracture. The Investigation for Maximal Peyronie’s Reduction Efficacy and Safety Studies (IMPRESS) I and II examined the clinical efficacy and safety of CCh intralesional injections in subjects with Peyronie disease. Primary outcomes in these identical phase 3 randomized, double blind, placebo controlled studies included the percent change in penile curvature and the change in the Peyronie disease questionnaire (PDQ) symptom bother score from baseline to 52 weeks. IMPRESS I and II examined CCh intralesional injections in 417 and 415 subjects, respectively, through a maximum of 4 treatment cycles, each separated by 6 weeks. Men received up to 8 injections of 0.58 mg CCh, comprising 2 injections per 6 week cycle separated by approximately 24 to 72 hours with subsequent penile plaque modeling within 24 to 72 hours. Men were stratified by baseline penile curvature (30 to 60 vs. 61 to 90 degrees) and randomized to CCh or placebo 2:1 in favour of the former. Post hoc meta-analysis of IMPRESS I and II data revealed that men treated with CCh showed a mean 34% improvement in penile curvature, representing a mean ±SD −17.0±14.8 degree change per subject, compared with a mean 18.2% improvement in placebo treated men, representing a mean −9.3±13.6 degree change per subject (P<0.0001). The mean change in PDQ symptom bother score was significantly improved in treated men vs men on placebo (−2.8±3.8 vs. −1.8±3.5, P=0.0037). Three serious adverse events (corporeal rupture) were surgically repaired. IMPRESS I and II support the clinical efficacy and safety of CCh for the physical and psychological aspects of Peyronie disease.

Virulence genes and clinical features of Clostridium difficile-associated diarrhea in children

Clostridium difficile is an obligate anaerobic Gram-positive bacillus, it can cause Clostridium difficile-associated diarrhea (CDAD). This study aimed to investigate the virulence genes and clinical features of CDAD in children by gene detection. From May 2012 to January 2013, the 121 inpatients in Beijing Children's Hospital who suffered from diarrhea after antibiotics treatment were detected for Clostridium difficile virulence genes including the five genes for pathogenic loci (tcdA, tcdB, tcdC, tcdD, tcdE) and the genes for binary toxin CDT (cdtA and cdtB) using polymerase chain reaction (PCR) in order to research the clinical features of CDAD, and analyze target products by sequencing. In the 121 children with diarrhea, 60 (49.6%) were toxin B-positive,including 12 toxin A-positive and toxin B-positive (A+B+), 48 toxin A-negative but toxin B-positive (A-B+). The toxin A-positive but toxin B-negative (A+B-) specimens or binary toxin (cdtA and cdtB)-positive specimens were not detected. Of 60 tcdB-positive specimens, tcdC, tcdD and tcdE positive specimens were 24 (40%), 25 (42%), 24 (40%), respectively. The sequencing results of tcdA, tcdB, tcdC, tcdD, and tcdE gene were consistent with the reference sequence. In the 60 children with CDAD, infants (≤3 years) accounted for 62% (37/60). The duration of diarrhea was 3-77 days, and 42 (70%) cases had acute diarrhea; 39 (65%) patients had fever, 40 (67%) had anemia, 36 (60%) had abnormal white blood cell count, 30 (50%) had hypoalbuminemia, 25 (42%) had elevated C-reactive protein (CRP). The level of CRP in positive group was significantly higher compared to the negative group (45.0(16.0,89.0) mg/L vs. 19.0(14.5,41.5) mg/L, Z=-2.008, P=0.045). The level of plasma albumin in positive group was significantly lower compared to the negative group (35.3(29.7,39.8) g/L vs. 38.5(33.9,41.5) g/L, Z=-2.610, P=0.009). There were no significant differences in gender, age, duration of diarrhea, hospital staytime, time of using antibiotics and laboratory test between A+B+ group and A-B+ group (all P>0.05). The main virulence genotype of Clostridium difficile was toxin A-negative but toxin B-positive in this research. The clinical features of CDAD in children were acute diarrhea with fever. Laboratory examination showed that white blood cell count was abnormal, CRP was increased, hemoglobin and plasma albumin were reduced.

Burden of Clostridium difficile infection in the United States.

for the CHIPS Study Group Since publication of their article, the authors report no further potential conflict of interest. 1. Lewis G, ed. Saving mothers’ lives: reviewing maternal deaths to make motherhood safer — 2003-2005: the seventh report on confidential enquiries into maternal deaths in the United Kingdom. London: The Confidential Enquiry into Maternal and Child Health (CEMACH), December 2007 (http://www.hqip.org.uk/ assets/NCAPOP-Library/CMACE-Reports/18.-December-2007 -Saving-Mothers-Lives-reviewing-maternal-deaths-to-make -motherhood-safer-2003-2005-Executive-Summary.pdf).

A sensitive method for detection of Clostridium difficile in diarrheal patients

A sensitive method for detection of Clostridium difficile in diarrheal patients

In vitro effects of elastase and collagenase on mechanical properties of hamster lungs.

A previous study of hamsters, 21 days after intratracheal treatment with pancreatic elastase, showed development of emphysema, shift of the volume-pressure curve up an to the left, with both air and saline filling, and increase in quasistatic lung compliance. There was also a striking increase in vital capacity and lung volume at transpulmonary pressure of 25 cm H2O (TLC25); however, 21 days after collagenase treatment, there was only a slight increase in TLC25. The lung volume changes were not consistent with the theory that the collagen fiber network is responsible for limiting distension of the lung. This report considers saline-filled volume-pressure curves studied in excised hamster lungs after incubation with endotracheally instilled pancreatic elastase or clostridial collagenase solutions. Fluid retained in the lungs after the first infusion-withdrawal cycle was significantly greater in lungs treated with elastase than in lungs treated with collagenase or in control lungs. Total fluid volume at full inflation was similar in the 3 groups. Chord compliance of lungs treated with collagenase was greater at high volume range than that in lungs treated with elastase or control lungs; chord compliance of elastase-treated lungs was higher at mid-volume range than that of collagenase-treated or control lungs. The results of these in vitro studies are consistent with the theory of independently functioning elastic and collagen fiber networks, with the latter limiting lung distensibility at high volumes, and the former providing great extensibility at low volumes. Events that are part of the repair process after elastase injury may result in a change in the orientation of collagen in alveolar tissue and appear to account for the differing effects of in vivo and in vitro elastase treatment on the static mechanical properties of the lungs.

Treatment with Nontoxigenic

Recurrent Clostridium difficile infection (CDI) appears to stem from the germination of toxigenic C. difficile spores in the intestinal tract in the

Research progress of Clostridium difficile-associated diarrhea

Clostridium difficile is a gram-positive, obligate anaerobic bacillus, which is one of the most common pathogenic bacteria of antibiotic associated diarrhea, and can cause Clostridium difficile-associated diarrhea.In recent years, the incidence of Clostridium difficile infection has increased significantly in the world with the excessive use of broad-spectrum antibiotics, the increase of strains resistance, and the emergence of hypervirulent strains.This paper presents a brief review on research progress of Clostridium difficile-associated diarrhea. Key words: Clostridium difficile; Toxin; Diarrhea; Infection; Child

Clostridium cost-of-illness studies use heterogeneous methods

Clostridium cost-of-illness studies use heterogeneous methods

Clinico-pathological findings of Clostridium perfringens type D enterotoxaemia in goats and its hemolytic activity in different erythrocytes.

The present investigation was conducted to study the effects of experimental Clostridium perfringens type D enterotoxaemia in teddy goats. Clinical signs started to appear after 30 min of experimental infection like anorexia, diarrhea, dehydration, frothing and dyspnea. Gross lesions consisted of severe congestion in tissues of varying intensity with enlarged mesenteric lymph nodes while histological examination revealed edema of lungs, kidney, and lymph nodes and to some extent in brain along with hemorrhages in lungs and intestines. Clostridium perfringens type D carrying alpha and epsilon toxin genes were amplified with amplicon size about 247 bp and 665 bp, respectively. Human erythrocytes showed the highest hemolysis, 68%, followed by mice, 57%, against culture supernatants. The percentage of hemolysis was significantly higher at 37°C as compared to 25°C except for rabbit and dog.

Progress in diagnosis and treatment of pseudomembranous colitis

Pseudomembranous colitis occurred is closely related to the use of antibiotics and the pathogenic bacteria approximately 100% is clostridium difficile.Diagnosis is mainly based on clinical manifestations, stool culture, toxin detection, colonoscopy and/or histological examination.Once the diagnosis has been confirmed or highly suspected, the initial antibiotic therapy should be discontinued, add with metronidazole and/or vancomycin, supplement by probiotic and nutritional support.The recent years, new treatment options have been proposed.This article presents an review of the related situation of pseudomembranous colitis to improve the comprehension for clinical doctors. Key words: Pseudomembranous colitis; Clostridium difficile; Diagnosis; Treatment