References of "Taminiau, Bernard"
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See detailLaboratory idenification 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 detailIdentification of Shiga toxin-producing (STEC) and enteropathogenic (EPEC) Escherichia coli in diarrhoeic calves and comparative genomics of O5 bovine and human STEC
Fakih, Ibrahim; Thiry, Damien ULg; Duprez, Jean-Noël ULg et al

in Veterinary Microbiology (2016)

Escherichia coli producing Shiga toxins (Stx) and the attaching-effacing (AE) lesion (AE-STEC) are responsible for (bloody) diarrhoea in humans and calves while the enteropathogenic E. coli (EPEC ... [more ▼]

Escherichia coli producing Shiga toxins (Stx) and the attaching-effacing (AE) lesion (AE-STEC) are responsible for (bloody) diarrhoea in humans and calves while the enteropathogenic E. coli (EPEC) producing the AE lesion only cause non-bloody diarrhoea in all mammals. The purpose of this study was (i) to identify the pathotypes of enterohaemolysin-producing E. coli isolated between 2009 and 2013 on EHLY agar from less than 2 month-old diarrhoeic calves with a triplex PCR targeting the stx1, stx2, eae virulence genes; (ii) to serotype the positive isolates with PCR targeting the genes coding for ten most frequent and pathogenic human and calf STEC O serogroups; and (iii) to compare the MLSTypes and virulotypes of calf and human O5 AE-STEC after Whole Genome Sequencing using two server databases (www.genomicepidemiology.org). Of 233 isolates, 206 were triplex PCR-positive: 119 AE-STEC (58%), 78 EPEC (38%) and 9 STEC (4%); and the stx1+eae+ AE-STEC (49.5%) were the most frequent. Of them, 120 isolates (84% of AE-STEC, 23% of EPEC, 22% of STEC) tested positive with one O serogroup PCR: 57 for O26 (47.5%), 36 for O111 (30%), 10 for O103 (8%) and 8 for O5 (7%) serogroups. The analysis of the draft sequences of 15 O5 AE-STEC could not identify any difference correlated to the host. As a conclusion, (i) the AE-STEC associated with diarrhoea in young calves still belong to the same serogroups as previously (O5, O26, O111) but the O103 serogroup may be emerging, (ii) the O5 AE-STEC from calves and humans are genetically similar [less ▲]

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See detailNo favorable effect of reduced tillage on microbial community diversity in a silty loam soil (Belgium)
Degrune, Florine ULg; Theodorakopoulos, Nicolas ULg; Dufrêne, Marc ULg et al

in Agriculture, Ecosystems & Environment (2016)

Among the soil management practices used to promote sustainable agriculture, reduced tillage and retention of residues from the previous crop are reported to enhance significantly both soil fertility and ... [more ▼]

Among the soil management practices used to promote sustainable agriculture, reduced tillage and retention of residues from the previous crop are reported to enhance significantly both soil fertility and crop productivity. Here, high-throughput sequencing (454 technology) was used to see how the tillage regime (conventional vs. reduced tillage) and the fate of crop residues (retention or removal) affect microbial communities at two sampling depths (top soil: 0–5 cm and deeper soil: 15–20 cm) in a fertile silty loam soil in Belgium. All combinations of these three factors were studied. After 6 years of conversion from conventional to reduced tillage, depth emerged as the main factor responsible for variation in microbial diversity, tillage regime ranked second, and finally, crop residue fate had no influence on microbial diversity. For both bacteria and fungi, the diversity appeared higher in the top soil than in the deeper soil, and surprisingly, higher under conventional than under reduced tillage. These differences are explained by changes in community composition due to taxon loss rather than taxon replacement. The specific local set of environmental conditions (a loess-derived soil and an oceanic temperate climate) may explain these results. These observations raise the question: does impoverishment in indicator taxa influence soil processes, and thus crop production? To answer this question, we discuss how the presence of certain indicator taxa liable to play an ecological role might relate to crop productivity. [less ▲]

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See detailAdding mucins to an in vitro batch fermentation model of the large intestine induces changes in microbial population isolated from porcine feces depending on the substrate
Tran, Thi Hanh Tham ULg; Boudry, Christelle ULg; Everaert, Nadia ULg et al

in FEMS Microbiology Ecology (2016), 92(2), 13

Adding mucus to in vitro fermentation models of the large intestine showed that some genera, namely lactobacilli, are dependent on host-microbiota interactions and that they rely on mucosa layers to ... [more ▼]

Adding mucus to in vitro fermentation models of the large intestine showed that some genera, namely lactobacilli, are dependent on host-microbiota interactions and that they rely on mucosa layers to increase their activity. This study investigates whether this dependence on mucus is substrate-dependent and to which extend other genera are impacted by the presence of mucus. Inulin and cellulose were fermented in vitro by a fecal inoculum from pig in the presence or not of mucin-beads in order to compare fermentation patterns and bacterial communities. Mucins increased final gas production with inulin and shifted short-chain fatty acids molar ratios (P<0.001). QPCR analyses revealed that Lactobacillus spp. and Bifidobacterium spp. decreased with mucins, but Bacteroides spp. increased when inulin was fermented. A more in-depth community analysis indicated that the mucins increased Proteobacteria (0.55 vs. 0.25 %, P=0.013), Verrucomicrobia (5.25 vs. 0.03 %, P=0.032), Ruminococcaceae, Bacteroidaceae, and Akkermansia spp.. Proteobacteria (5.67 vs. 0.55 %, P<0.001) and Lachnospiraceae (33 vs. 10.4 %) were promoted in the mucuscompared to the broth, while Ruminococcaceae decreased. The introduction of mucins affected many microbial genera and fermentation patterns, but from PCA results, the impact of mucus was independent from the fermentation substrate. [less ▲]

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See detailExploring the bacterial diversity of Belgian steak tartare using metagenetics and qPCR analysis
Delhalle, Laurent ULg; Korsak Koulagenko, Nicolas ULg; Taminiau, Bernard ULg et al

in Journal of Food Protection (2016), 79(2), 200-229

Steak tartare is a popular meat dish in Belgium. It is prepared with raw ground minced beef and eaten with sauce, vegetables, and spiced. Since it contains raw meat, steak tartare is highly prone to ... [more ▼]

Steak tartare is a popular meat dish in Belgium. It is prepared with raw ground minced beef and eaten with sauce, vegetables, and spiced. Since it contains raw meat, steak tartare is highly prone to bacterial spoilage. The objective of this study was to explore the bacterial flora diversity in steak tartare in Belgium according to the source and to determine which bacteria are able to grow during the shelf life. A total of 58 samples from butchers’ shops, restaurants, sandwich shops and supermarkets were collected. These samples were analyzed using 16S rDNA metagenetics, a classical microbiological technique, and quantitative real-time PCR (qPCR) targeting the Lactobacillus genus. Samples were analyzed at the beginning and at the end of their shelf life, except for those from restaurants and sandwich shops analyzed only at the purchase date. Metagenetic analysis identified up to 180 bacterial species and 90 genera in some samples. But only seven bacterial species were predominant in the samples, depending on the source: Brochothrix thermosphacta, Lactobacillus algidus, Lactococcus piscium, Leuconostoc gelidum, Photobacterium kishitani, Pseudomonas spp. and Xanthomonas oryzae. With this work, an alternative method is proposed to evaluate the total flora in food samples based on the number of reads from metagenetic analysis and the results of qPCR. The degree of underestimation of aerobic plate counts (APCs) at 30°C estimated with the classical microbiology method was demonstrated in comparison with the proposed culture independent method. Compared to culture-based methods, metagenetic analysis combined with qPCR targeting Lactobacillus provides valuable information for characterizing the bacterial flora of raw meat. [less ▲]

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See detailNo favorable effect of reduced tillage on microbial communities in a silty loam soil (Belgium)
Degrune, Florine ULg; Theodorakopoulos, Nicolas ULg; Dufrêne, Marc ULg et al

Poster (2015, December 01)

To date, only a few studies have applied metagenomics to investigate the influence of different tillage regimes and types of crop residue management on soil microbial communities. These studies were ... [more ▼]

To date, only a few studies have applied metagenomics to investigate the influence of different tillage regimes and types of crop residue management on soil microbial communities. These studies were conducted under specific climates on soils characterized by particular land-use histories. A very different ecological context is to be found in certain areas of Western Europe, such as central Belgium, whose loess-derived soils are among the most fertile in the world and have long been used for intensive agriculture. Specific objectives were to determine diversity levels and changes in microbial community composition under different combinations of tillage regime (conventional vs. reduced) and crop residue fate (residue removal R- vs. residues left R+ on the field). As reduced tillage results in two contrasting zones (the first centimeters of soil are mixed each year, while the soil below remains unperturbed), we chose to perform the analysis at two depths: 0 to 5 cm and 15 to 20 cm. [less ▲]

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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 detailMetagenomic analysis of samples
Daube, Georges ULg; Taminiau, Bernard ULg; Nezer, Carine et al

Patent (2015)

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See detailUse of 16S rDNA Metagenetics and classical Microbiology to Assess the bacterial superficial Contamination Patterns in Bovines Classically Slaughtered or following the Halal Ritual
Korsak Koulagenko, Nicolas ULg; Taminiau, Bernard ULg; Hupperts, Caroline et al

Poster (2015, June 17)

In Belgium and in several European countries, two cattle slaughtering protocols exist: the classical method, that encompasses a stunning step before the sticking procedure, and the halal method, combining ... [more ▼]

In Belgium and in several European countries, two cattle slaughtering protocols exist: the classical method, that encompasses a stunning step before the sticking procedure, and the halal method, combining the stunning and the sticking in one single step. The main difference lies in the fact that, in the halal protocol, a single cut with a sharp knife is practiced directly on live cattle, instead of two cutting steps with two different knives for the sticking in the classical slaughtering technique. The unique section in the halal technique results generally in the cross section of trachea and esophagus of cattle. The aim of this study was to seek if the two slaughtering techniques were similar regarding the superficial contamination of carcasses, swabbed between 2 and 4 hours after the killing step. For this purpose, classical microbiological tests (TVC and Enterobacteriaceae) and 16S rDNA metagenetic analysis were carried out from 20 cattle carcasses (swabbing of “legal” zone – 1.600 cm2 – and in the neck area – 200 cm2). The classical microbiological results revealed no significant differences between the two slaughtering practices. Statistical analysis of pyrosequencing data showed that differences in bacterial population abundance between slaughtering methods were mainly found in the “legal” swabbing zone compared to the neck area. Bacterial genera belonging to Actinobacteria (Brevibacterium, Corynebacterium) were more aundant in “Halal” samples whereas populations from the Proteobacteria (Caulobacteraceae, Comamonadaceae, Bradyrhizobiaceae) and Firmicutes (Lactobacillus) were more abundand in the “classical” group. The analysis of OTU abundance of bacteria from the digestive or respiratory tract revealed no differences beteween groups. In conclusion, the slaughtering method does not influence the superficial microbiological pattern in terms of specific microbiological markers of the digestive or respiratory tract. However, precise analysis to the genus level underlines differences between methods, the legal swabbing zone being still the best sampling zone compared to the neckline. The next step will be the identification of precise contamination origin of the differences found between slaughtering methods. [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 detailSoil microbial community composition changes according to the tillage practice and plant development stage
Degrune, Florine ULg; Dufrêne, Marc ULg; Taminiau, Bernard ULg et al

Poster (2015, April)

Agricultural practices have a strong impact on soil bacterial and fungal community composition. Furthermore, microbial community composition can change with the stage of plant development. We are ... [more ▼]

Agricultural practices have a strong impact on soil bacterial and fungal community composition. Furthermore, microbial community composition can change with the stage of plant development. We are interested in exploring these effects in relation to changes induced by agriculture (conventional and reduced tillage) and plant stage (germination and flowering) in soil conditions. Here, instead of examining this impact at a high taxonomic level such as phylum and/or class, thus missing potentially relevant information from lower levels, we propose an original method: exploiting the available sequence information at the lowest taxonomic level attainable for each operational taxonomic unit. Results show that some microbial communities were impacted only by the tillage practice , while others were impacted only by the stage of plant. Changes in microbial community composition could be due to the soil conditions induced by the soil practice and the stage of plant. [less ▲]

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See detailA novel sub-phylum method discriminates better the impact of crop management on soil microbial community
Degrune, Florine ULg; Dufrêne, Marc ULg; Colinet, Gilles ULg et al

in Agronomy for Sustainable Development (2015)

Soil microorganisms such as mycorrhizae and plant-growth-promoting rhizobacteria have beneficial effects on crop productivity. Agricultural practices are known to impact soil microbial communities, but ... [more ▼]

Soil microorganisms such as mycorrhizae and plant-growth-promoting rhizobacteria have beneficial effects on crop productivity. Agricultural practices are known to impact soil microbial communities, but past studies examining this impact have focused mostly on one or two taxonomic levels, such as phylum and class, thus missing potentially relevant information from lower levels. Therefore we propose here an original, sub-phylum method for studying how agricultural practices modify microbial communities. This method involves exploiting the available sequence information at the lowest taxonomic level attainable for each operational taxonomic unit. In order to validate this novel method we assessed microbial community composition using 454 pyrosequencing of 16S and 28S rRNA genes, then we compared the results with results of a phylum-level analysis. Agricultural practices included conventional tillage, reduced tillage, residue removal and residue retention. Results show that, at the lowest taxonomic level attainable, tillage is the main factor influencing both bacterial community composition, accounting for 13% of the variation, and fungal community composition, accounting for 18% of the variation. Whereas phylum-level analysis failed to reveal any effect of soil practice on bacterial community composition, and missed the fact that different members of the same phylum responded differently to tillage practice. For instance, the fungal phylum Chytridiomycota showed no impact of soil treatment, while sub-phylum-level analysis revealed an impact of tillage practice on the Chytridiomycota sub-groups Gibberella, which includes a notorious wheat pathogen, and Trichocomaceae. This clearly demonstrates the necessity of exploiting the information obtainable at sub-phylum level when assessing the effects of agricultural practice on microbial communities. [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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