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See detailClostridium difficile a new zoonotic agent? Assessment of human transmission potential of hypervirulent strains of Clostridium difficile through food products consumption
Rodriguez Diaz, Cristina ULg

Doctoral thesis (2016)

Clostridium difficile is a spore-forming anaerobic bacterium recognised as a major human pathogen responsible for pseudomembranous colitis and nosocomial-antibiotic associated diarrhea. Traditionally ... [more ▼]

Clostridium difficile is a spore-forming anaerobic bacterium recognised as a major human pathogen responsible for pseudomembranous colitis and nosocomial-antibiotic associated diarrhea. Traditionally, hospitals were considered the main reservoirs for infection. However, in the last years the incidence, deaths, complications and costs of C. difficile infection (CDI) have been rising, not only in healthcare facilities, but also within the community. In the community, it has been detected in a growing number of CDI cases in previously healthy individuals without antimicrobial exposure, hospital stay or any other classical risk factors. Furthermore, the disease has been repeatedly described in younger patients, including children. Some hypotheses have been proposed to explain this peak of community cases, the most obvious being that nowadays more attention is given to CDI surveillance. In the last years, diarrhea due to C. difficile disease might have gone undiagnosed, and in many cases went unreported, particularly in the community. Since 2004, severe outbreaks of CDI have been documented increasingly in the United States, Canada and in Europe. These outbreaks have been associated with the emergence of a novel strain, known as PCR-ribotype 027, characterised by higher than usual levels of toxins A and B production, and the presence of a third toxin named CDT or binary toxin. This strain type is also characterised by its resistance to both erythromycin and fluoroquinolones (i.e. moxifloxacin, gatifloxacin and levofloxacin). However, according to the latest hospital surveillance studies in Europe, since 2010 there is a decrease in the incidence rates of PCR-ribotype 027 while other PCR-ribotypes, including PCR-ribotypes 014, 020, 001, 002, 078 and 015 are increasing. Person to person contact is one source proposed for the spread in the community, occurring after visiting hospitalised patients or residents in long-term care facilities. Employees of these health-care settings can also carry spores and contaminate their entourage. The second hypothesis is contamination from the environment, following visits to a potentially contaminated place, such as hospitals or nursing homes. The two most important potential sources of CDI in the community, which have been demonstrated by investigations in the last decade are animals and foods. While C. difficile is also known as enteric pathogen in some food producing and companion animal species, there are several reports describing the presence of the bacterium in the intestinal contents of apparently healthy animals. Moreover, data published recently suggests animals as an important source of human CDI, which can spread disease through environmental contamination, direct or indirect contact, or food contamination, including carcass and meat contamination at slaughter or in the case of crops, through the use of organic animal manure. By definition, zoonoses are infectious diseases that can be transmitted directly or indirectly between animals and humans, through direct contact or close proximity with infected animals, or through the environment. Foodborne zoonotic pathogens are transmitted via the consumption of contaminated food or drink water. The first description of C. difficile in domestic animals and their environments dates from 1974 and possible foodborne transmission was reported for the first time in 1982. However, nowadays the importance of C. difficile as zoonotic disease remains largely unknown. The "One Health" concept is a worldwide strategy, which recognises that the health of humans and animals is connected and also depends on the environment. The present dissertation is a 5 year national study that has investigated the presence of C. difficile in animals and food, from “farm-totable”. The study was also extended to humans resident in a nursing home and in two hospitals in Belgium and in Spain. The characterisation of the isolates obtained has ultimately allowed comparison of the PCR-ribotype distribution in the different European hospitals, as well as with the PCR-ribotype distribution found in animals and foods. This work explores how C. difficile spreads among human patients, animals, foods and the environment to better understand the potential of the bacterium as a zoonotic or foodborne infectious agent. [less ▲]

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See detailLaboratory identification of anaerobic bacteria isolated on Clostridium difficile selective medium
Rodriguez Diaz, Cristina ULg; Warszawski, Nathalie; Korsak Koulagenko, Nicolas ULg et al

in Acta Microbiologica et Immunologica Hungarica (2016)

Despite increasing interest in the bacterium, the methodology for Clostridium difficile recovery has not yet been standardised. Cycloserine cefoxitin fructose taurocholate (CCFT) has historically been the ... [more ▼]

Despite increasing interest in the bacterium, the methodology for Clostridium difficile recovery has not yet been standardised. Cycloserine cefoxitin fructose taurocholate (CCFT) has historically been the most used medium for C. difficile isolation from human, animal, environmental and food samples, and presumptive identification is usually based on colony morphologies. However, CCFT is not totally selective. This study describes the recovery of 24 bacteria species belonging to 10 different genera other than C. difficile, present in the environment and foods of a retirement establishment that were not inhibited in the C. difficile selective medium. These findings provide insight for further environmental and food studies as well as for isolation of C. difficile on supplemented CCFT. [less ▲]

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See detailAssociation of Targeted metagenomic analysis and classical microbiology for Clostridium difficile detection and microbial ecosystem mapping of surfaces hands and foodstuffs in a meat processing plant
Rodriguez Diaz, Cristina ULg; Oukbir, Meriem; Taminiau, Bernard ULg et al

Poster (2016, May 11)

INTRODUCTION Zoonoses are infectious that can be transmitted between animals and humans through direct contact, close proximity or the environment. Since domestic and food animals frequently test positive ... [more ▼]

INTRODUCTION Zoonoses are infectious that can be transmitted between animals and humans through direct contact, close proximity or the environment. Since domestic and food animals frequently test positive for the bacterium, it seems plausible that C. difficile could be zoonotic. PURPOSE This study aimed to determine the prevalence and the epidemiology of C. difficile in cattle farms and the possible spread of the bacterium among animals and farmers. METHODS A total of 176 faecal samples of cattle were collected from 5 different Belgian farms. A stool sample of each farmer was also requested. Detection of C. difficile was performed by classical culture on C. difficile selective medium (cycloserine cefoxitin fructose cholate). Isolates were characterised by PCR-ribotyping and Genotype Cdiff test (Hain Lifescience), which allows the detection of all toxin genes, mutations in gyrA gene and the deletion in the regulator gene tcdC. RESULTS C. difficile was detected in 14/178 (7.9%) animal samples. Isolates were grouped into five different types, including PCR-ribotype 015. All of them were identified as toxigenic. In contrast, none of the 5 farmers studied were positive for the bacterium. SIGNIFICANCE Results obtained indicate that PCR-ribotypes commonly isolated from hospitalised patients are also present in cattle, indicating an animal reservoir. However, a zoonotic transmission could be not demonstrated in the farms studied. [less ▲]

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See detailClostridium difficile in Food and Animals: A Comprehensive Review.
Rodriguez Diaz, Cristina ULg; Taminiau, Bernard ULg; Van Broeck, J. et al

in Advances in Experimental Medicine and Biology (2016)

Zoonoses are infections or diseases that can be transmitted between animals and humans through direct contact, close proximity or the environment. Clostridium difficile is ubiquitous in the environment ... [more ▼]

Zoonoses are infections or diseases that can be transmitted between animals and humans through direct contact, close proximity or the environment. Clostridium difficile is ubiquitous in the environment, and the bacterium is able to colonise the intestinal tract of both animals and humans. Since domestic and food animals frequently test positive for toxigenic C. difficile, even without showing any signs of disease, it seems plausible that C. difficile could be zoonotic. Therefore, animals could play an essential role as carriers of the bacterium. In addition, the presence of the spores in different meats, fish, fruits and vegetables suggests a risk of foodborne transmission. This review summarises the current available data on C. difficile in animals and foods, from when the bacterium was first described up to the present. [less ▲]

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See detailClostridium difficile presence in Spanish and Belgian hospitals
Rodriguez Diaz, Cristina ULg; Fernadez, Jonathan; Van Broeck, Johan et al

in Microbial Pathogenesis (2016), 100

Clostridium difficile is recognised worldwide as the main cause of infectious bacterial antibiotic-associated diarrhoea in hospitals and other healthcare settings. The aim of this study was to first ... [more ▼]

Clostridium difficile is recognised worldwide as the main cause of infectious bacterial antibiotic-associated diarrhoea in hospitals and other healthcare settings. The aim of this study was to first survey C. difficile prevalence during the summer of 2014 at the Central University Hospital of Asturias (Spain). By typing the isolates obtained, it was then possible to compare the ribotype distribution at the Spanish hospital with results from the St Luc University Hospital in Belgium over the same period. The prevalence of positive cases reported in Spain and Belgium was 12.3% and 9.3% respectively. The main PCR-ribotypes previously described in Europe were found in both hospitals, including 078, 014, 012, 020 and 002. In the Spanish hospital, most of the C. difficile-positive samples were referred from oncology, acute care and general medicine services. In the Belgian hospital the majority of positive samples were referred from the paediatric service. However, a high percentage of isolates from this service were non-toxigenic. This study finds that the presence and detection of C. difficile in paediatric and oncology services requires further investigation. [less ▲]

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See detailLongitudinal survey of Clostridium difficile presence and gut microbiota composition in a Belgian nursing home
Rodriguez Diaz, Cristina ULg; Taminiau, Bernard ULg; Korsak Koulagenko, Nicolas ULg et al

in BMC Microbiology (2016), 16(229),

ackground Increasing age, several co-morbidities, environmental contamination, antibiotic exposure and other intestinal perturbations appear to be the greatest risk factors for C. difficile infection (CDI ... [more ▼]

ackground Increasing age, several co-morbidities, environmental contamination, antibiotic exposure and other intestinal perturbations appear to be the greatest risk factors for C. difficile infection (CDI). Therefore, elderly care home residents are considered particularly vulnerable to the infection. The main objective of this study was to evaluate and follow the prevalence of C. difficile in 23 elderly care home residents weekly during a 4-month period. A C. difficile microbiological detection scheme was performed along with an overall microbial biodiversity study of the faeces content by 16S rRNA gene analysis. Results Seven out of 23 (30.4 %) residents were (at least one week) positive for C. difficile. C. difficile was detected in 14 out of 30 diarrhoeal samples (43.7 %). The most common PCR-ribotype identified was 027. MLVA showed that there was a clonal dissemination of C. difficile strains within the nursing home residents. 16S-profiling analyses revealed that each resident has his own bacterial imprint, which was stable during the entire study. Significant changes were observed in C. difficile positive individuals in the relative abundance of a few bacterial populations, including Lachnospiraceae and Verrucomicrobiaceae. A decrease of Akkermansia in positive subjects to the bacterium was repeatedly found. Conclusions A high C. difficile colonisation in nursing home residents was found, with a predominance of the hypervirulent PCR-ribotype 027. Positive C. difficile status is not associated with microbiota richness or biodiversity reduction in this study. The link between Akkermansia, gut inflammation and C. difficile colonisation merits further investigations. [less ▲]

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See detailClostridium difficile infection: Early history, diagnosis and molecular strain typing methods.
Rodriguez Diaz, Cristina ULg; Van Broeck, Johan; Taminiau, Bernard ULg et al

in Microbial Pathogenesis (2016), 97

Recognised as the leading cause of nosocomial antibiotic-associated diarrhoea, the incidence of Clostridium difficile infection (CDI) remains high despite efforts to improve prevention and reduce the ... [more ▼]

Recognised as the leading cause of nosocomial antibiotic-associated diarrhoea, the incidence of Clostridium difficile infection (CDI) remains high despite efforts to improve prevention and reduce the spread of the bacterium in healthcare settings. In the last decade, many studies have focused on the epidemiology and rapid diagnosis of CDI. In addition, different typing methods have been developed for epidemiological studies. This review explores the history of C. difficile and the current scope of the infection. The variety of available laboratory tests for CDI diagnosis and strain typing methods are also examined. [less ▲]

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See detailStudy of the Potential Zoonotic Transmission of Clostridium difficile in Belgian Cattle Farms.
Rodriguez Diaz, Cristina ULg; Hakimi, Djalal-Eddine; Daube, Georges ULg et al

in Journal of Food Protection (2016), 79(supplement A),

Introduction: Zoonoses are infectious that can be transmitted between animals and humans through direct contact, close proximity or the environment. Since domestic and food animals frequently test ... [more ▼]

Introduction: Zoonoses are infectious that can be transmitted between animals and humans through direct contact, close proximity or the environment. Since domestic and food animals frequently test positive for the bacterium, it seems plausible that C. difficile could be zoonotic. A former study showed that the prevalence in veal calf aged less than 6 months was 22% while in adult cattle population, it was 6,9 %. Purpose: This study aimed to determine the prevalence and the epidemiology of C. difficile in cattle farms and the possible spread of the bacterium among animals and farmers. Methods: A total of 176 faecal samples of cattle were collected from 5 different Belgian farms (south East Belgium), from November 2015 to February 2016. A stool sample of each farmer was also requested. Detection of C. difficile was performed by classical culture on C. difficile selective medium (cycloserine cefoxitin fructose cholate). Isolates were characterised by PCR-ribotyping and Genotype Cdiff test (Hain Lifescience), which allows the detection of all toxin genes, mutations in gyrA gene and the deletion in the regulator gene tcdC. Toxic activity was confirmed by a cytotoxic assay on MRC-5 cells. Results: C. difficile was detected in 14/178 (7.9%) animal samples. Isolates were grouped into five different types, including PCR-ribotype 015 (this ribotype is one the most encountered in hospitals in Belgium). The other types were UCL46A, UCL24*, UCL24, UCL33. All of them were identified as toxigenic by cytotoxicity assay and toxin genes profile. In contrast, none of the 5 farmers studied were positive for the bacterium. Significance: Results obtained indicate that PCR-ribotypes commonly isolated from hospitalised patients are also present in cattle, indicating an animal reservoir. However, a zoonotic transmission could be not demonstrated in this preliminary study. [less ▲]

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See detailClostridium difficile in foods and animals: A comprehensive review
Rodriguez Diaz, Cristina ULg; Taminiau, Bernard ULg; Van Broeck, Johan et al

in Advances in Experimental Medicine and Biology (2016), 27

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See detailFaecal microbiota characterisation of horses using 16 rdna barcoded pyrosequencing, and carriage rate of clostridium difficile at hospital admission
Rodriguez Diaz, Cristina ULg; Taminiau, Bernard ULg; Brévers, Bastien et al

in BMC Microbiology (2015), 15

Background The equine faecal microbiota is very complex and remains largely unknown, while interspecies interactions have an important contribution to animal health. Clostridium difficile has been ... [more ▼]

Background The equine faecal microbiota is very complex and remains largely unknown, while interspecies interactions have an important contribution to animal health. Clostridium difficile has been identified as an important cause of diarrhoea in horses. This study provides further information on the nature of the bacterial communities present in horses developing an episode of diarrhoea. The prevalence of C. difficile in hospitalised horses at the time of admission is also reported. Results Bacterial diversity of the gut microbiota in diarrhoea is lower than that in non-diarrhoeic horses in terms of species richness (p-value <0.002) and in population evenness (p-value: 0.02). Statistical differences for Actinobacillus, Porphyromonas, RC9 group, Roseburia and Ruminococcaceae were revealed. Fusobacteria was found in horses with diarrhoea but not in any of the horses with non-diarrheic faeces. In contrast, Akkermansia was among the three predominant taxa in all of the horses studied. The overall prevalence of C. difficile in the total samples of hospitalised horses at admission was 3.7 % (5/134), with five different PCR-ribotypes identified, including PCR-ribotype 014. Two colonised horses displayed a decreased bacterial species richness compared to the remaining subjects studied, which shared the same Bacteroides genus. However, none of the positive animals had diarrhoea at the moment of sampling. Conclusions The abundance of some taxa in the faecal microbiota of diarrhoeic horses can be a result of microbiome dysbiosis, and therefore a cause of intestinal disease, or some of these taxa may act as equine enteric pathogens. Clostridium difficile colonisation seems to be transient in all of the horses studied, without overgrowth to trigger infection. A large proportion of the sequences were unclassified, showing the complexity of horses’ faecal microbiota. [less ▲]

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See detailClostridium difficile, a new zoonotic agent. Characterization and relatedness of C. difficile strains isolated from animals, food and humans in Belgium
Rodriguez Diaz, Cristina ULg

Poster (2015, July 28)

Introduction: Clostridium difficile is an anaerobic spore-forming bacterium recognised as a major cause of nosocomial colitis and antibiotic associated diarrhea. Over the past few years, several studies ... [more ▼]

Introduction: Clostridium difficile is an anaerobic spore-forming bacterium recognised as a major cause of nosocomial colitis and antibiotic associated diarrhea. Over the past few years, several studies have focused on the possible role of animals and food as contamination routes for human C. difficile infections. Purpose: The aim of this study was to isolate and compare the C. difficile strains circulating in animals, food and humans in Belgium. Methods: Faecal samples of newborn pigs and calves were collected from breeding farms. Intestinal contents and carcasses samples were collected from cattle and pigs at slaughterhouse. Raw meat was obtained from the retail trade. Horse faecal samples were collected from hospitalized animals. Human C. difficile isolates were obtained from care home residents and hospitalized patients. C. difficile strains were compared with respect to the toxin gene profile, PCR-ribotyping, antimicrobial activity, multilocus sequence typing (MLST) and multiple-locus variable number tandem repeat analysis (MLVA). A neighbourd-joining phylogenetic three was constructed in order to determine the correlation between human, animal and food isolates. Results: A total of 127 isolates belonging to 32 different PCR-ribotypes were collected. The PCR-ribotypes most prevalent in terms of number of isolates were 078, 014 and 027. For a given PCR-ribotype, strains presented a similar susceptibility to the antimicrobials tested, irrespective of the isolation source. Phylogenetic analysis showed that human, meat and animal isolates with the same PCR-ribotype cluster in the same lineage. Significance: The overlap between strains from animal, food and human origins suggest a potential risk of interspecies and foodborne transmission. [less ▲]

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See detailCARRIAGE OF CLOSTRIDIUM DIFFICILE IN HOSPITAL PATIENTS IN SPAIN, INCLUDING MOLECULAR CHARACTERIZATION AND ANTIMICROBIAL SUSCEPTIBILITY OF THE ISOLATES
Rodriguez Diaz, Cristina ULg; Taminiau, Bernard ULg; Van Broeck, Johan et al

Poster (2015, May 21)

Increasing age, several co-morbidities, environmental contamination, antibiotic exposure and other intestinal perturbations appear to be the greatest risk factors for C. difficile infection (CDI ... [more ▼]

Increasing age, several co-morbidities, environmental contamination, antibiotic exposure and other intestinal perturbations appear to be the greatest risk factors for C. difficile infection (CDI). Therefore, hospitalized patients are considered particularly vulnerable to CDI. The main objective of this study was to evaluate the prevalence of C. difficile in a Spanish hospital and to characterize the isolates with respect to the PCR-ribotype, antibiotic resistance and toxin activity. In addition, multilocus sequence typing (MLST) was performed in order to study clonal relationships between C. difficile isolates obtained from two different countries. Culture of samples was performed in a selective medium cycloserine cefoxitin fructose cholate. An identification of the isolated colonies was done by PCR detection of tpi, tcdA, tcdB and cdtA genes. Toxic activity was confirmed by a cytotoxic immunoassay. Further characterization was performed by PCR ribotyping. MLST was used in order to determine genetic relationships between Spanish and Belgian C. difficile isolates recovered from hospital patients in both countries. C. difficile was frequently detected in hospitalized patients. The isolates belonged to different PCR ribotypes, including type 027. Most of the strains obtained harboured tcdA and tcdB genes. The number of positive faecal samples considerably increased among elderly patients over 65 years old. The multi-locus sequence typing (MLST) analysis showed that some Spanish and Belgian isolates cluster in the same lineage. This study reveals the circulation of toxigenic C. difficile in a Spanish hospital. The relatedness between Belgian and Spanish isolates indicate a common source or a lack of diversity from some PCR-ribotypes. Moreover, data obtained suggest that the combination of ribotyping and MLST is a good tool for the inter-laboratory comparison of strains among different countries. Continuous inter-country surveillance is suitable to understand the spread of C. difficile isolates among hospitalized patients. [less ▲]

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See detailHeat survival of Clostridium difficile spores in ground meat during cooking process
Rodriguez Diaz, Cristina ULg; Taminiau, Bernard ULg; Avesani, Véronique et al

Poster (2015, April 21)

Introduction: Clostridium difficile is a spore-forming pathogen considered as a major cause of enteric disease in humans, with faecal-oral route as the primary mode of transmission. However, recent ... [more ▼]

Introduction: Clostridium difficile is a spore-forming pathogen considered as a major cause of enteric disease in humans, with faecal-oral route as the primary mode of transmission. However, recent studies have reported the occurrence of C. difficile in ground meats at retail stores, indicating that foods could be an additional source of infection in the community. Purpose: The objective of this study was to determine the resistance of C. difficile spores in contaminated ground meat during cooking process. Methods: Prior to testing, to obtain spores and to enhance heterogeneity, spores of two different strains were produced in two nutritious broths. C. difficile spores were experimentally inoculated in 45 g of ground meat (beef and pork) in order to obtain a final contamination of 4,500 ufc g-1. Six heating temperatures (70, 75, 80, 85, 90 and 95°C) were chosen. Samples were heating in a water bath with an integrated program for time-temperature. One sample without inoculum was used as control with a temperature probe placed inside. Once the desired temperature was research in the core of the sample, the heat treatment was prolonged for 10 min. Subsequently, all the samples were placed on the chilling room (4°C) before analyse. These experiments were conducted in duplicate with a spore enumeration in triplicate. Results: Heating contaminated ground meat at 70, 75 and 80°C for 10 min was not effective for C. difficile spores inhibition. However, 10 min of heat shock at 80°C was the only temperature that significantly reduced the number of countable colonies. Heat treatment at 85°C (or more) inhibits the germination of both of the strains tested. Significance: Ensure that ground meat, like burgers or sausages, is heated to more than 85°C would be an important measure to reduce the risk of C. difficile food transmission. [less ▲]

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See detailAssociation of classical microbiology and 16S rDNA metagenetic analysis to evaluate the presence of Clostridium difficile ina a belgian nursing home
Rodriguez Diaz, Cristina ULg; Taminiau, Bernard ULg; Avesani, Véronique et al

Poster (2015, April 01)

Increasing age, several co-morbidities, environmental contamination, antibiotic exposure and other intestinal perturbations appear to be the greatest risk factors for C. difficile infection (CDI ... [more ▼]

Increasing age, several co-morbidities, environmental contamination, antibiotic exposure and other intestinal perturbations appear to be the greatest risk factors for C. difficile infection (CDI). Therefore, elderly care home residents are considered particularly vulnerable to CDI. The main objective of this study was to evaluate and follow the prevalence of C. difficile in a Belgian nursing home. During a 4-month period, stool samples from a group of 23 elderly care home residents were collected weekly. A C. difficile microbiological detection scheme was performed along with an overall microbial biodiversity study of the faeces content by Targeted Metagenomic analysis. Culture of samples was performed in a selective medium cycloserine cefoxitin fructose cholate. An identification of the isolated colonies was done by PCR detection of tpi, tcdA, tcdB and cdtA genes. Toxic activity was confirmed by a cytotoxic immunoassay. Further characterization was performed by PCR ribotyping. The Metagenomic analysis was targeted on the v1-v3 hyper-variable region of 16S rDNA. The taxonomical assignment of the populations was performed with MOTHUR and Blast algorithms. Seven out of 23 (30.4%) residents were (at least one week) positive for C. difficile. The most common PCR-ribotype identified was 027. Targeted Metagenomic analyses reveals that each resident has his own bacterial imprint, which is stable during the entire study. Residents’ positives for C. difficile by classical microbiology showed an important proportion of C. difficile sequences. However, Metagenomics analysis can’t substitute targeted protocols. It was not used as a diagnostic tool to detect C. difficile but rather to determine the identification and correlations of the major bacterial populations that are present in the gut microbiota. In conclusion, this unique association of classical microbiology protocol with pyrosequencing allowed to follow C. difficile in patients and to identify several other bacterial populations whose abundance is correlated with C. difficile. [less ▲]

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See detailClostridium difficile from food and surface samples in a Belgian nursing home: An unlikely source of contamination
Rodriguez Diaz, Cristina ULg; Korsak Koulagenko, Nicolas ULg; Taminiau, Bernard ULg et al

in Anaerobe (2015), 32

This study investigates the contamination of foods and surfaces with Clostridium difficile in a single nursing home. C. difficile PCR-ribotype 078 was found in one food sample and in none of the tested ... [more ▼]

This study investigates the contamination of foods and surfaces with Clostridium difficile in a single nursing home. C. difficile PCR-ribotype 078 was found in one food sample and in none of the tested surfaces. These results indicate that food and surfaces are an unlikely source of C. difficile infection in this setting. [less ▲]

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See detailL’exploration du microbiote intestinal par les outils de la métagénomique au service de la santé animale et humaine
Taminiau, Bernard ULg; Rodriguez Diaz, Cristina ULg; Delhalle, Laurent et al

in Food Science and Law (2015), (4), 165-169

L’essor des nouvelles technologies de séquençage de l’ADN a, depuis une décennie, bouleversé les méthodologies et ouvert de nouvelles perspectives de recherche dans de nombreux domaines. La métagénétique ... [more ▼]

L’essor des nouvelles technologies de séquençage de l’ADN a, depuis une décennie, bouleversé les méthodologies et ouvert de nouvelles perspectives de recherche dans de nombreux domaines. La métagénétique et la métagénomique, appliquées à la microbiologie, a permis de changer la vision du monde scientifique face à l’omniprésence des populations microbiennes dans notre quotidien. Basées sur l’exploration de la biodiversité taxonomique et fonctionnelle des bactéries, de nombreuses recherches ayant pour but le contrôle et le management des écosystèmes microbiens ont vu le jour, dans le domaine de la santé, dans l’industrie mais également dans la gestion de notre environnement. Cette revue détaille et illustre brièvement les techniques métagénomiques actuelles. [less ▲]

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See detailInvestigation of clostridium difficile interspecies relatedness using multilocus sequence typing, multilocus variable-number tandem-repeat analysis and antimicrobial susceptibility testing
Rodriguez Diaz, Cristina ULg; Avesani, Véronique; Taminiau, Bernard ULg et al

in Veterinary Journal (2015), 206

Multilocus sequence typing (MLST), multilocus variable-number tandem-repeat analysis (MLVA) and antimicrobial susceptibility were performed on 37 animal and human C. difficile isolates belonging to 15 ... [more ▼]

Multilocus sequence typing (MLST), multilocus variable-number tandem-repeat analysis (MLVA) and antimicrobial susceptibility were performed on 37 animal and human C. difficile isolates belonging to 15 different PCR-ribotypes in order to investigate the relatedness of human and animal isolates and to identify possible transmission routes. MLVA identified a total of 21 different types while MLST only distinguished 12 types. Identical C. difficile strains were detected in the same animal species for PCR-ribotypes 014, 078, UCL 16U and UCL 36, irrespective of their origin or the isolation date. Non clonal strains were found among different hosts; however, a high genetic association between pig and cattle isolates belonging to PCR-ribotype 078 was revealed. MLVA also showed genetic differences that clearly distinguished human from animal strains. For a given PCR-ribotype, human and animal strains presented a similar susceptibility to the antimicrobials tested. All strains were susceptible to vancomycin, metronidazole, chloramphenicol and rifampicin, while PCR-ribotypes 078, UCL 5a, UCL 36 and UCL 103 were associated with erythromycin resistance. The data suggest a wide dissemination of clones at hospitals and breeding-farms or a contamination at the slaughterhouse, but less probability of interspecies transmission. However, further highly discriminatory genotyping methods are necessary to elucidate interspecies and zoonotic transmission of C. difficile. [less ▲]

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See detailClostridium difficile infection and intestinal microbiota interactions
Rodriguez Diaz, Cristina ULg; Taminiau, Bernard ULg; Van Broeck, Johan et al

in Microbial Pathogenesis (2015), 89

Clostridium difficile remains the leading cause of healthcare-associated diarrhoea and outbreaks continue to occur worldwide. Aside from nosocomial C. difficile infection, the bacterium is also ... [more ▼]

Clostridium difficile remains the leading cause of healthcare-associated diarrhoea and outbreaks continue to occur worldwide. Aside from nosocomial C. difficile infection, the bacterium is also increasingly important as a community pathogen. Furthermore, asymptomatic carriage of C. difficile in neonates, adults and animals is also well recognised. The investigation of the gut's microbial communities, in both healthy subjects and patients suffering C. difficile infection (CDI), provides findings and information relevant for developing new successful approaches for its treatment, such as faecal microbiota transplantation, or for the prophylaxis of the infection by modification of the gut microbiota using functional foods and beverages. The analysis of all available data shows new insights into the role of intestinal microbiota in health and disease. [less ▲]

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