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See detailMicrobiological risks and benefits of the consumption of raw milk and the effect of heat treatment
Verraes, Claire; Cardoen, S.; Claeys, W. et al

Conference (2014, September 01)

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See detailMetagenomic insights into the dynamics and functionality of food microbial communities
Champomier Vergès, Marie-Christine; Daube, Georges ULg

Conference (2014, September)

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See detailModelling the development of potential spoilage and biopreservative microorganisms on precooked pasta in different conditions of temperature based on classical microbiology and 16S rDNA metagenomic
Gand, Mathieu ULg; Cargnel, Mickaël ULg; Kergourlay, Gilles ULg et al

Poster (2014, September)

An important way to prevent the spoilage of food is the respect of the cold chain during the storage. While the temperature instructions are generally respected during process and distribution of food ... [more ▼]

An important way to prevent the spoilage of food is the respect of the cold chain during the storage. While the temperature instructions are generally respected during process and distribution of food products, it is not always the case with the consumers. Indeed, a few persons reach the right temperature level required for a safe storage of foodstuffs in their refrigerator. Besides, the food can sometimes spend a few hours in ambient temperature between the buying in the supermarket and the storage in cold temperature. In this study, we propose to model the growth of microorganisms on precooked pasta, stored in different conditions of temperature that reflect the situations described above (constant 4°C, constant 8°C, constant 12°C, 1/3 4°C – 2/3 8°C, 1/3 4°C – breach during 4h at 20°C – 2/3 8°C and 1/3 8°C – breach during 4h at 20°C – 2/3 8°C. The product was surface inoculated with potential spoilage and biopreservative strains (Lactococcus piscium, Leuconostoc mesenteroides and Leuconostoc citreum). Analyses by classical microbiology and V1-V3 16S rDNA metagenomics were done each day until the out of date of the food matrix. The transition from 4 to 8°C and the breach at 20°C during 4h have clearly boosted the growth of the microorganisms. The metagenomic analysis was a powerful tool to follow separately each population in each condition of storage. The results of this communication show the importance of keeping the foodstuffs in 4°C or lower in the refrigerator with the goal to avoid the spoilage or the development of pathogens and the potential of metagenomics for selection of biopreservative strains. [less ▲]

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See detailMultilocus sequence typing analysis and antibiotic resistance of Clostridium difficile strains isolated from retail meat and humans in Belgium
Rodriguez Diaz, Cristina ULg; Taminiau, Bernard ULg; Avesani, Véronique et al

in Food Microbiology (2014), 42

Clostridium difficile has been isolated from food animals and meat, specially ground pork and ground beef. The recovered isolates were closely related to C. difficile human strains, indicating that ... [more ▼]

Clostridium difficile has been isolated from food animals and meat, specially ground pork and ground beef. The recovered isolates were closely related to C. difficile human strains, indicating that animals and food are possible transmission routes of human C. difficile infection. The main objective of this study was to characterize C. difficile isolates from retail meat and to compare with human isolates recovered from hospital patients in Belgium. Raw meat (beef and pork) was obtained from the retail trade. C. difficile was recovered from 2.3% of the beef samples and from 4.7% of the pork samples. A total of 4 different PCR-ribotypes were identified with a large percentage of types 078 and 014. Resistance to moxifloxacin and erythromycin was detected. The multi-locus sequence typing (MLST) analysis showed that meat and human isolates cluster in the same lineage. This study reveals the presence of toxigenic C. difficile in retail meat in Belgium with predominance PCR-ribotypes 078 and 014, which are among the four most prevalent ribotypes of C. difficile isolated from humans in Europe. [less ▲]

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See detailModelling the development of potential spoilage and biopreservative microorganisms on white pudding in different conditions of temperature based on classical microbiology and 16S rDNA metagenomic
Gand, Mathieu ULg; Cauchie, Emilie ULg; Kergourlay, Gilles ULg et al

Poster (2014, September)

An important way to prevent the spoilage of food is the respect of the cold chain during the storage. While the temperature instructions are generally respected during process and distribution of food ... [more ▼]

An important way to prevent the spoilage of food is the respect of the cold chain during the storage. While the temperature instructions are generally respected during process and distribution of food products, it is not always the case with the consumers. Indeed, a few persons reach the right temperature level required for a safe storage of foodstuffs in their refrigerator. Besides, the food can sometimes spend a few hours in ambient temperature between the buying in the supermarket and the storage in cold temperature. In this study, we propose to model the growth of microorganisms on white pudding, stored in different conditions of temperature that reflect the situations described above (constant 4°C, constant 8°C, constant 12°C, 1/3 4°C – 2/3 8°C, 1/3 4°C – breach during 4h at 20°C – 2/3 4°C and 1/3 4°C – breach during 4h at 20°C – 2/3 8°C. The product was surface inoculated with potential spoilage and biopreservative strains (Raoultella terrigena, Serratia quinivorans, Carnobacterium maltaromaticum, Lactobacillus oligofermentans, Lactobacillus nenjiangensis, Lactobacillus fuchuensis, Leuconostoc mesenteroides, lactococcus lactis and Lactobacillus graminis). Analyses by classical microbiology and V1-V3 16S rDNA metagenomics were done each day until the out of date of the food matrix. The transition from 4 to 8°C and the breach at 20°C during 4h have clearly boosted the growth of the microorganisms. The metagenomic analysis was a powerful tool to follow separately each population in each condition of storage. The results of this communication show the importance of keeping the foodstuffs in 4°C or lower in the refrigerator with the goal to avoid the spoilage or the development of pathogens and the potential of metagenomics for selection of biopreservative strains. [less ▲]

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See detailCarnobacterium maltaromaticum isolated from vacuum-packed beef with long shelf life: morphological and functional characterization
Didimo Imazaki, Pedro Henrique ULg; Tahiri, Assia ULg; Ndedi Ekolo, François et al

Poster (2014, September)

This study was conducted as part of a research project on meat conservability, in particular vacuum packaged beef displaying very long shelf lives at a temperature close to the freezing point, and its ... [more ▼]

This study was conducted as part of a research project on meat conservability, in particular vacuum packaged beef displaying very long shelf lives at a temperature close to the freezing point, and its objective was to perform a morphological and functional characterization of C. maltaromaticum with potential bioprotective effect isolated from vacuum packaged long shelf life beef. The isolated C. maltaromaticum strain presented similar morphological, biochemical and enzymatic profiles as those of two reference strains (LMG 11393 and LMG 22902). Among the studied conditions, a temperature of +12 °C and an atmosphere poor in oxygen (100 % N2) were the optimal conditions for the growth of C. maltaromaticum. Nevertheless, growth on lower temperatures is also possible. After inoculation of beef samples with C. maltaromaticum and 7 days of storage under vacuum at −1 °C, no effect was observed on the total viable count and on the count of lactic acid bacteria. A reduction of Pseudomonas sp. and Brochothrix thermosphacta was observed during the first week of storage under vacuum conditions. After 7 days of subsequent storage of the same samples under modified atmosphere at +4 °C, the inoculant favored the growth of B. thermosphacta and inhibited the growth of Enterobacteriaceae under 100 % N2. No effect of the inoculant was observed when an atmosphere rich in oxygen was applied. The evaluation of the influence of different atmospheres showed that the growth of C. maltaromaticum was slower in an atmosphere containing O2 and CO2. Long-term storage under vacuum at low temperatures are therefore suitable for the growth of this bacterium. An antimicrobial effect against Enterobacteriaceae was highlighted on inoculated fresh meat stored under N2. The functional characterization of this strain will be further pursued by genotypic characterization. Special attention will be taken to study its bioprotective properties. [less ▲]

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See detailCarriage and acquisition rates of Clostridium difficile in hospitalized horses, including molecular characterization, multilocus sequence typing and antimicrobial susceptibility of bacterial isolates
Rodriguez Diaz, Cristina ULg; Taminiau, Bernard ULg; Brévers, Bastien et al

in Veterinary Microbiology (2014), 172

lostridium difficile has been identified as a significant agent of diarrhoea and enterocolitis in both foals and adult horses. Hospitalization, antibiotic therapy or changes in diet may contribute to the ... [more ▼]

lostridium difficile has been identified as a significant agent of diarrhoea and enterocolitis in both foals and adult horses. Hospitalization, antibiotic therapy or changes in diet may contribute to the development of C. difficile infection. Horses admitted to a care unit are therefore at greater risk of being colonized. The aim of this study was to investigate the carriage of C. difficile in hospitalized horses and the possible influence of some risk factors in colonization. During a seven-month period, faecal samples and data relating the clinical history of horses admitted to a veterinary teaching hospital were collected. C. difficile isolates were characterized through toxin profiles, cytotoxicity activity, PCR-ribotyping, antimicrobial resistance and multilocus sequence typing (MLST). Ten isolates were obtained with a total of seven different PCR-ribotypes, including PCR-ribotype 014. Five of them were identified as toxinogenic. A high resistance to gentamicin, clindamycin and ceftiofur was found. MLST revealed four different sequencing types (ST), which included ST11, ST26, ST2 and ST15, and phylogenetic analysis showed that most of the isolates clustered in the same lineage. Clinical history suggests that horses frequently harbour toxigenic and non-toxigenic C. difficile and that in most cases they are colonized regardless of the reason for hospitalization; the development of diarrhoea is more unusual. [less ▲]

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See detailAntimicrobial and molecular analysis of Salmonella serovar Livingstone strains isolated from humans in Tunisia and Belgium
Guedda, Intissar; Taminiau, Bernard ULg; Ferjani, Asma et al

in Journal of Infection in Developing Countries [=JIDC] (2014), 8

Introduction: Salmonella Livingstone is one of the most common serotypes responsible for nosocomial outbreaks in Tunisia. In this study, 42 isolates of Salmonella Livingstone were analyzed. Most of these ... [more ▼]

Introduction: Salmonella Livingstone is one of the most common serotypes responsible for nosocomial outbreaks in Tunisia. In this study, 42 isolates of Salmonella Livingstone were analyzed. Most of these were isolated from humans (31 strains from Tunisia and 9 strains from Belgium) and 2 isolates came from food products (beef and pork). Methodology: All strains were characterized by antibiogram, multilocus sequence typing (MLST), and virulotyping. This last technique was carried out by simple PCR of five chromosomal genes (agfA, hin/H2, iroB, phoP/Q, and slyA) and two plasmid genes (spvA and spvC). Results: All Tunisian strains were resistant to amoxicillin, amoxicillin-clavulanic acid, ticarcillin, cefalotin, gentamicin, and kanamycin. They were also resistant to third-generation cephalosporin antibiotics (cefotaxim and ceftazidim). Belgian isolates were susceptible to all antibiotics tested. Further to MLST analyses, Tunisian strains belonged to the same sequence type, ST543. For Belgian isolates, eight strains had a ST543 profile, two strains had a ST638 profile, and one strain had a ST457 profile. Analyses of the virulence gene contents showed that strains isolated in different years and from different origins had the same virulence profile. These carried all five chromosomal genes and lacked plasmid-located virulence genes spvA and spvC. Conclusions: A combination of different typing methods showed that the majority of Belgian strains and all Tunisian strains were closely related; they belonged to the same sequence type (ST543) and had the same virulence profile, but different antibiotic resistance profiles depended on the country of origin. [less ▲]

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See detailMorphological and functional characterization of Carnobacterium maltaromaticum isolated from vacuum-packed beef with long shelf life
Didimo Imazaki, Pedro Henrique ULg; Tahiri, Assia ULg; Ndedi Ekolo, François et al

Poster (2014, August)

The aim of this study was to perform a morphological and functional characterization of a Carnobacterium maltaromaticum strain with a potential bioprotective effect isolated from vacuum packaged long ... [more ▼]

The aim of this study was to perform a morphological and functional characterization of a Carnobacterium maltaromaticum strain with a potential bioprotective effect isolated from vacuum packaged long shelf life beef. The morphological, biochemical and enzymatic profiles, the influence of different temperatures and atmospheres, and the microbial stability of fresh beef inoculated with the C. maltaromaticum strain were evaluated. The isolated C. maltaromaticum strain presented similar morphological, biochemical and enzymatic profiles as those of two reference strains (LMG 11393 and LMG 22902). The growth of C. maltaromaticum was slower in an atmosphere containing O2 and CO2. Vacuum packing is therefore suitable for this bacterium. An antimicrobial effect against Enterobacteriaceae was highlighted on inoculated fresh meat stored under N2. The functional characterization of this isolate will be further pursued by a genotypic characterization to better understand its potential bioprotective effect. [less ▲]

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See detailProduction and microbiological evaluation of three types of "Dèguè", a local fermented drink made from milk in Benin
Tchekessi, Célestin; Bokossa, Auréole; Agbangla, Clément et al

in International Journal of Multidisciplinary and Current Research (2014), 2

This study consists to finalize some technologies for the production of a fermented drink called dèguè. This drink is widely consumed in Benin and other countries in sub-Saharan Africa. Following three ... [more ▼]

This study consists to finalize some technologies for the production of a fermented drink called dèguè. This drink is widely consumed in Benin and other countries in sub-Saharan Africa. Following three different technologies, we had produced three (03) types of dèguè respectively with maize flour, sorghum and millet. These types have been analyzed and their microbiological characteristics were evaluated. The microbiological analysis results obtained from the experiments have shown that lactic acid bacteria, yeasts and molds were the dominant microflora of dèguè and varied respectively 7.22log10UFC/g to 7.55log10UFC/g for lactic acid bacteria and 7.78log10UFC/g to 8.44log10UFC/g for yeasts and molds. Moreover, the statistical analysis of these results showed that there was no significant difference at 5% (p <0.05) between the three types of dèguè. The values of lactic acid bacteria obtained were consistent with the standard (≥ 107/g) attached to the yoghurt. No type contained neither total coliforms nor thermotolerant coliforms. The products (dèguès) were also free of pathogenic microorganisms such as Escherichia coli, Staphylococcus aureus and Salmonella. [less ▲]

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See detailHigh throughput sequencing analysis reveals genetic variability and selection pressure in different murine norovirus genomic regions during in vitro replication
Mauroy, Axel ULg; Taminiau, Bernard ULg; Nezer, Carine et al

Poster (2014, July)

Murine norovirus (MuNoV), a single stranded positive sense RNA virus belonging to the Caliciviridae family, is considered as a representative model for human norovirus infections, one of the most ... [more ▼]

Murine norovirus (MuNoV), a single stranded positive sense RNA virus belonging to the Caliciviridae family, is considered as a representative model for human norovirus infections, one of the most important etiological cause of both epidemic and sporadic gastroenteritis cases worldwide. Four open reading frames are described into its genome: ORF1 codes the non-structural (NS) proteins, including the viral RNA dependent RNA polymerase (RdRp); ORF2 codes the single capsid protein (VP1), wherein two domains are present: a relatively conserved domain (“shell”) and a more variable domain (“protruding”); ORF3 codes a minor structural protein; and ORF4, currently only found in viruses genetically related to MuNoV codes a virulence factor. In this study, we demonstrated by high throughput sequencing that, during serial passages of MuNoV in cell culture, the substitution rates, estimated by Bayesian inferences, did not significantly differ across the five targeted genomic regions except one. These rates were similar in four genomic regions encompassing partial non-structural 1-2 protein (NS1-2)-, NS5-, NS6-, NS7 (RdRp)- and VP1-coding sequences (coding the conserved part of the protein also including the ORF4 region). In the partial minor structural protein-coding region, this substitution rate was however estimated to be at least one log higher when expressed as substitution/site/day. The precise localisation of the detected nucleotide point mutations (substitution, deletion and insertion) were reported as well as the quantitative increase or decrease of the sequences harbouring them along ten cell culture passages. The non-silent amino acid mutations were also depicted in 3D models for four out of the five studied regions. These results have important implications for different norovirus research fields, especially in terms of diagnosis, classification methodology and genetic evolution. [less ▲]

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See detailXylo-oligosaccharides improve broiler performance by stimulating butyrate production by Clostridium cluster XIVa strains
De Maesschalck, C.; Nezer, Carine; Taminiau, Bernard ULg et al

Conference (2014, June 16)

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See detailComparison and molecular characterization of animal and human Clostridium difficile strains
Rodriguez Diaz, Cristina ULg; Taminiau, Bernard ULg; Korsak Koulagenko, Nicolas ULg et al

Poster (2014, May 07)

The main objective of this study was to characterize and compare animal and human C. difficile strains with respect to the PCR-ribotype and the antibiotic resistance. Multilocus sequence typing analysis ... [more ▼]

The main objective of this study was to characterize and compare animal and human C. difficile strains with respect to the PCR-ribotype and the antibiotic resistance. Multilocus sequence typing analysis (MLST) was performed in order to study clonal relationships of the isolates. [less ▲]

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See detailStudy of the microbial flora of steak tartare by metagenomic approach
Korsak Koulagenko, Nicolas ULg; Delhalle, Laurent ULg; Nezer, Carine et al

Poster (2014, May 06)

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See detailL'analyse par séquençage à haut débit révèle la variabilité génétique et la pression de sélection dans différentes régions génomiques du norovirus murin durant sa réplication in vitro
Mauroy, Axel ULg; Taminiau, Bernard ULg; Nezer, Carine et al

Conference (2014, March)

Le norovirus murin (MuNoV), un virus à ARN de polarité positive appartenant à la famille des Caliciviridae, est considéré comme un modèle adéquat pour les infections humaines à norovirus, une des causes ... [more ▼]

Le norovirus murin (MuNoV), un virus à ARN de polarité positive appartenant à la famille des Caliciviridae, est considéré comme un modèle adéquat pour les infections humaines à norovirus, une des causes étiologiques les plus importantes dans les cas de gastroentérite épidémique ou sporadique dans le monde entier. Quatre cadres de lecture ouverts (ORF) sont décrits au sein de son génome : l’ORF1 code les protéine non structurales (NS), dont l’ARN polymérase ARN dépendante virale (RdRp) ; l’ORF2 code l’unique protéine de capside (VP1), dans laquelle sont décrites deux régions : une relativement conservée (domaine « shell ») et une autre beaucoup plus variable (domaine « protruding ») ; l’ORF3 code une protéine structurale mineure ; et l’ORF4, actuellement uniquement décrit chez les virus génétiquement apparentés au MuNoV, code un facteur de virulence. Dans cette étude, nous démontrons par séquençage à haut débit que, durant des passages successifs du MuNoV en culture cellulaire, les taux de substitution, estimés par inférences Bayésiennes, n’ont pas significativement différé au travers des cinq régions génomiques ciblées à l’exception d’une région bien précise. Ces taux étaient similaires pour quatre régions englobant des séquences partielles codant les protéines non structurales NS1-2, NS5, NS6 et NS7 (RdRp) et VP1 dans sa région conservée (incluant également l’ORF4). Dans la région codant partiellement la protéine structurale mineure, ce taux de substitution, exprimé en substitution/site/jour, a été cependant estimé être plus élevée d’au moins une unité logarithmique. La localisation précise des mutations ponctuelles détectées (substitution, délétion et insertion) est rapportée ainsi que l’augmentation ou la diminution quantitative du nombre des séquences qui les présentaient au cours de dix passages successifs en culture cellulaire. Les localisations des mutations non silencieuses ont aussi été représentées dans une modélisation tridimensionnelle de quatre des cinq régions étudiées. Ces résultats ont d’importantes implications pour différents champs de recherche sur les norovirus, spécialement en termes de diagnostic, de méthodologie de classification et d’évolution génétique. [less ▲]

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See detailChemical composition and antimicrobial activity of essential oils of Ocimum basilicum, Ocimum canum and Ocimum gratissimum in function of harvesting time
Mith, Hasika; Yayi, Éléonore; Bokossa, Innocent et al

in Journal of Essential Oil-Bearing Plants [=JEOBP] (2014)

BACKGROUND: The aim of present the study was to investigate the chemical composition and the antimicrobial activity of essential oils of Ocimum basilicum, Ocimum canum and Ocimum gratissimum from Benin as ... [more ▼]

BACKGROUND: The aim of present the study was to investigate the chemical composition and the antimicrobial activity of essential oils of Ocimum basilicum, Ocimum canum and Ocimum gratissimum from Benin as affected by harvesting time. The chemical composition of hydrodistillated essential oils were analyzed by GC-FID (gas chromatography – flame ionization detector) and GC-MS (gas chromatography-mass spectrometry). Disc diffusion and broth microdilution assays were used to evaluate the antibacterial activity of essential oils against two foodborne pathogens. RESULTS: Based on the composition analysis, major components were as follows: estragol (43.0 -44.7 %) and linalool (24.6 -29.8 %) in O. basilicum oils; carvacrol (12.0 -30.8 %) and p-cymene (19.5 -26.2 %) in O. canum oils; thymol (28.3 -37.7 %) and γ-terpinene (12.5 -19.3 %) in O. gratissimum oils. The tested oils and their components exhibited notable antimicrobial activities against Listeria monocytogenes and Salmonella typhimurium. The O. canum and O. gratissimum oils collected at 7h and 19h showed significant higher activities against L. monocytogenes and S. typhimurium (MICs and MBCs 0.34 – 2.5 µL/mL) (p < 0.05), whereas O. basilicum showed lower activity (MICs and MBCs 2.0 – 8.0 µL/mL) at any daytime of harvest, the weakest being at 19h (MIC and MBC 12.0 – 32.0 µL/mL). CONCLUSION: The daytime of harvest can influence the composition of oils and their activities on bacteria. [less ▲]

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See detailA review of the microbiological hazards of raw milk from animal species other than cows
Verraes, Claire; Claeys, Wendy; Cardoen, Sabine et al

in International Dairy Journal (2014), 39

This review concentrates on information concerning the microbiological hazards that can be present in raw milk from animal species other than cows. Total bacterial counts of raw milk are described for ... [more ▼]

This review concentrates on information concerning the microbiological hazards that can be present in raw milk from animal species other than cows. Total bacterial counts of raw milk are described for several animal species, indicating the quality of the milk, then frequencies of occurrence of several human pathogenic microorganisms are considered and, finally, human cases of illness and outbreaks due to the consumption of raw milk from non-bovine species are covered. Only raw milk from goats and camels has so far been reported to be associated with outbreaks. Raw milk from horse and donkey may have a higher microbiological quality than raw milk from other animal species, although human pathogenic strains of Streptococcus are considered as a microbiological hazard for such milk. For raw milk from other animal species, the main microbiological hazards seem to be human pathogenic Escherichia coli, Campylobacter spp., tick-borne encephalitis virus and Brucella spp. [less ▲]

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See detailAntimicrobial activities of commercial essential oils and their components against food-borne pathogens and food spoilage bacteria.
Mith, Hasika; Dure, Remi; Delcenserie, Véronique ULg et al

in Food science & nutrition (2014), 2(4), 403-16

This study was undertaken to determine the in vitro antimicrobial activities of 15 commercial essential oils and their main components in order to pre-select candidates for potential application in highly ... [more ▼]

This study was undertaken to determine the in vitro antimicrobial activities of 15 commercial essential oils and their main components in order to pre-select candidates for potential application in highly perishable food preservation. The antibacterial effects against food-borne pathogenic bacteria (Listeria monocytogenes, Salmonella Typhimurium, and enterohemorrhagic Escherichia coli O157:H7) and food spoilage bacteria (Brochothrix thermosphacta and Pseudomonas fluorescens) were tested using paper disk diffusion method, followed by determination of minimum inhibitory (MIC) and bactericidal (MBC) concentrations. Most of the tested essential oils exhibited antimicrobial activity against all tested bacteria, except galangal oil. The essential oils of cinnamon, oregano, and thyme showed strong antimicrobial activities with MIC >/= 0.125 muL/mL and MBC >/= 0.25 muL/mL. Among tested bacteria, P. fluorescens was the most resistant to selected essential oils with MICs and MBCs of 1 muL/mL. The results suggest that the activity of the essential oils of cinnamon, oregano, thyme, and clove can be attributed to the existence mostly of cinnamaldehyde, carvacrol, thymol, and eugenol, which appear to possess similar activities against all the tested bacteria. These materials could be served as an important natural alternative to prevent bacterial growth in food products. [less ▲]

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See detailClostridium difficile infection in elderly nursing home residents.
Rodriguez Diaz, Cristina ULg; Korsak Koulagenko, Nicolas ULg; Taminiau, Bernard ULg et al

in Anaerobe (2014), 30

Age-related changes in intestinal flora and host defences, the receipt of antibiotic treatment, and the presence of underlying diseases are some of the most common risk factors associated with Clostridium ... [more ▼]

Age-related changes in intestinal flora and host defences, the receipt of antibiotic treatment, and the presence of underlying diseases are some of the most common risk factors associated with Clostridium difficile infection. Therefore, retirement care facilities for elderly people have been pinpointed as frequent sources of contamination. There is only limited data regarding the presence and epidemiology of C. difficile in nursing homes, and this gap in the current literature emphasises the need to gain a better understanding of the situation in order to prevent the emergence of new outbreaks among this population group. [less ▲]

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