Center dEconomie Rurale

Rue, Belgium

Center dEconomie Rurale

Rue, Belgium
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Tretzel L.,German Sport University Cologne | Thomas A.,German Sport University Cologne | Geyer H.,German Sport University Cologne | Delahaut P.,Center dEconomie Rurale | And 2 more authors.
Analytical and Bioanalytical Chemistry | Year: 2015

Abstract Dried blood spot (DBS) sampling, a technique used for taking whole blood samples dried on a filter paper, was initially reported in 1963 by Robert Guthrie. While the diagnostic analysis of metabolic disorders in newborns was the focus of investigations at that time, the number of established applications for preclinical drug development, toxicological studies, and therapeutic drug monitoring increased enormously in the last decades. As a consequence of speed, simplicity, and minimal invasiveness, DBS recommends itself as the preferential technique in sports drug testing. The present approach highlights for the first time the development of a screening assay for the analysis of the synthetic human adrenocorticotropic hormone tetracosactide hexaacetate (Synacthen®) in DBS using liquid chromatography tandem mass spectrometry. Highly purified sample extracts were obtained by an advanced sample preparation procedure including the addition of an internal standard (d8-tetracosactide) and immunoaffinity purification. The method's overall recovery was 27.6 %, and the assay's imprecision was calculated between 8.1 and 17.9 % for intraday and 12.9 to 20.5 % for interday measurements. Stability of the synthetic peptide in DBS was shown for at least 10 days at room temperature and presents a major benefit, since a rapid degradation in conventionally applied matrices such as urine or plasma is well known. With a limit of detection of 50 pg/mL, a detection window of several hours is expected considering reported steady-state plasma levels of 300 pg/mL after intramuscular application of Synacthen® Depot (1 mg). The analysis of authentic DBS samples within the scope of an administration study with 250 μg Synacthen® (short stimulation test) demonstrated the great potential of the developed assay to simplify the analysis of Synacthen® for doping control purposes. © 2015 Springer-Verlag Berlin Heidelberg.


Grant
Agency: European Commission | Branch: FP7 | Program: CP-IP | Phase: KBBE-2007-2-4-02 | Award Amount: 7.51M | Year: 2008

RASFF alerts show that monitoring of chemical contaminants in food and feed is very relevant in European food safety. Also consumers placed chemical contaminants on top of the worry-scale of food-related risks. According to the General Food Law, food and feed industries are responsible for the safety of their products. Often expensive instrumental single-analyte methods are being applied by regulatory and industrial laboratories. There is an urgent need for replacement by validated screening tools which are simple, inexpensive and rapid, but also show multiplex capability by detecting as many contaminants in parallel as possible. The CONffIDENCE proposal has been designed to provide long-term solutions to the monitoring of persistent organic pollutants, perfluorinated compounds, pesticides, veterinary pharmaceuticals (coccidiostats, antibiotics), heavy metals and biotoxins (alkaloids, marine toxins, mycotoxins) in high-risk products such as fish and fish feed, cereal-based food/feed and vegetables. A balanced mix of novel multiplex technologies will be utilized, including dipsticks, flow cytometry with functionalised beads, SPR optical and electrochemical biosensors, cytosensors and metabolomics-like comprehensive profiling. After validation, the simplified methods will be applied in impact demonstrators that contribute to exposure assessment and validation of hazard models. Moreover, hazards of emerging contaminants will be assessed through toxicological testing. Dissemination to scientists and to relevant stakeholders, including the food and feed industry, regulatory control bodies, DG-SANCO, EFSA, exporting countries, CRLs, routine laboratories, CEN and consumers will be assured by e-communication, press releases, public workshops, open days, presentations, publications and a science education module. The consortium consists of 17 partners from 10 countries, representing 9 research institutes, 5 universities, 2 large food and feed industries and 1 SME.


Grant
Agency: European Commission | Branch: FP7 | Program: CP-FP | Phase: KBBE-2009-2-4-01 | Award Amount: 4.05M | Year: 2010

The NanoLyse project will focus on the development of validated methods and reference materials for the analysis of engineered nano-particles (ENP) in food and beverages. The developed methods will cover all relevant classes of ENP with reported or expected food and food contact material applications, i.e. metal, metal oxide/silicate, surface functionalised and organic encapsulate (colloidal/micelle type) ENP. Priority ENPs have been selected out of each class as model particles to demonstrate the applicability of the developed approaches, e.g. nano-silver, nano-silica, an organically surface modified nano-clay and organic nano-encapsulates. Priority will be given to methods which can be implemented in existing food analysis laboratories. A dual approach will be followed. Rapid imaging and screening methods will allow the distinction between samples which contain ENP and those that do not. These methods will be characterised by minimal sample preparation, cost-efficiency, high throughput and will be achieved by the application of automated smart electron microscopy imaging and screening techniques in sensor and immunochemical formats. More sophisticated, hyphenated methods will allow the unambiguous characterisation and quantification of ENP. These will include elaborate sample preparation, separation by flow field fractionation and chromatographic techniques as well as mass spectrometric and electron microscopic characterisation techniques. The developed methods will be validated using the well characterised food matrix reference materials that will be produced within the project. Small-scale interlaboratory method performance studies and the analysis of a few commercially available products claiming or suspect to contain ENP will demonstrate the applicability and soundness of the developed methods.


McGrath T.F.,Queen's University of Belfast | Buijs J.,General Electric | Huet A.C.,Center dEconomie Rurale | Delahaut P.,Center dEconomie Rurale | And 2 more authors.
Sensors and Actuators, B: Chemical | Year: 2013

Multiplexed immunochemical detection platforms offer the potential to decrease labour demands, increase sample throughput and decrease overall time to result. A prototype four channel multiplexed high throughput surface plasmon resonance biosensor was previously developed, for the detection of food related contaminants. A study focused on determining the instruments performance characteristics was undertaken. This was followed by the development of a multiplexed assay for four high molecular weight proteins. The protein levels were simultaneously evaluated in serum samples of 10-week-old veal calves (n = 24) using multiple sample preparation methods. Each of the biosensor's four channels were shown to be independent of one another and produced multiplexed within run repeatability (n = 6) ranging from 2.0 to 6.7%CV, for the four tested proteins, whilst between run reproducibility (n = 4) ranged from 1.5 to 8.9%CV. Four calibration curves were successfully constructed before serum sample preparation was optimised for each protein. Multiplexed concentration analysis was successfully performed on four channels revealing that each proteins concentration was consistent across the twenty-four tested animals. Signal reproducibility (n > 19) on a further long term study revealed coefficient of variation ranging from 1.1% to 7.3% and showed that the multiplexed assay was stable for at least 480 cycles. These findings indicate that the performance characteristics fall within the range of previously published data for singleplex optical biosensors and that the multiplexing biosensor is fit-for-purpose for simultaneous concentration analysis in many different types of applications such as the multiplexed detection of markers of growth-promoter abuse and multiplexed detection of residues of concern in food safety. © 2013 Elsevier B.V.


McNamee S.E.,Queen's University of Belfast | Elliott C.T.,Queen's University of Belfast | Delahaut P.,Center dEconomie Rurale | Campbell K.,Queen's University of Belfast
Environmental Science and Pollution Research | Year: 2013

A multiplex surface plasmon resonance (SPR) biosensor method for the detection of paralytic shellfish poisoning (PSP) toxins, okadaic acid (and analogues) and domoic acid was developed. This method was compared to enzyme-linked immunosorbent assay (ELISA) methods. Seawater samples (n = 256) from around Europe were collected by the consortia of an EU project MIcroarrays for the Detection of Toxic Algae (MIDTAL) and evaluated using each method. A simple sample preparation procedure was developed which involved lysing and releasing the toxins from the algal cells with glass beads followed by centrifugation and filtering the extract before testing for marine biotoxins by both multi-SPR and ELISA. Method detection limits based on IC20 values for PSP, okadaic acid and domoic acid toxins were 0.82, 0.36 and 1.66 ng/ml, respectively, for the prototype multiplex SPR biosensor. Evaluation by SPR for seawater samples has shown that 47, 59 and 61 % of total seawater samples tested positive (result greater than the IC20) for PSP, okadaic acid (and analogues) and domoic acid toxins, respectively. Toxic samples were received mainly from Spain and Ireland. This work has demonstrated the potential of multiplex analysis for marine biotoxins in algal and seawater samples with results available for 24 samples within a 7 h period for three groups of key marine biotoxins. Multiplex immunological methods could therefore be used as early warning monitoring tools for a variety of marine biotoxins in seawater samples. © 2012 Springer-Verlag Berlin Heidelberg.


Vandenberge V.,Belgium Institute for Agricultural and Fisheries Research | Delezie E.,Belgium Institute for Agricultural and Fisheries Research | Huyghebaert G.,Belgium Institute for Agricultural and Fisheries Research | Delahaut P.,Center dEconomie Rurale | And 3 more authors.
Food Additives and Contaminants - Part A Chemistry, Analysis, Control, Exposure and Risk Assessment | Year: 2012

In the poultry industry, the widespread use of veterinary drugs such as antimicrobial compounds may lead to the presence of residues in whole eggs, egg white and egg yolk. During this study, laying hens received experimental feed containing sulfadiazine or doxycycline at cross-contamination levels of 2.5%, 5% and 10% of the therapeutic concentration. Since the therapeutic dose is 250 mg kg-1 for both substances, cross-contamination concentrations in the feed of 6.25, 12.5 and 25 mg kg-1 were expected. Whole egg, egg white and egg yolk samples were collected during the treatment and depletion period and were analysed via liquid chromatography-tandem mass spectrometry. For both drugs, a plateau phase was reached within 3-5 days and residue concentrations were detected in all egg matrices. For the 10% cross-contamination group, residual sulfadiazine concentrations of 208, 299 and 60 μg kg-1 and residual doxycycline concentrations of 455, 332, 206 μg kg-1 were detected in whole egg, egg white and egg yolk on day 13 of the treatment period, respectively. Both sulfadiazine and doxycycline had higher concentrations in egg white than in egg yolk, but the egg white-egg yolk ratio was higher for sulfadiazine than for doxycycline. As neither drug is allowed in Belgium for use in laying hens, residues may pose food safety concerns. © 2012 Copyright Taylor and Francis Group, LLC.


Vandenberge V.,Belgium Institute for Agricultural and Fisheries Research | Delezie E.,Belgium Institute for Agricultural and Fisheries Research | Huyghebaert G.,Belgium Institute for Agricultural and Fisheries Research | Delahaut P.,Center dEconomie Rurale | And 2 more authors.
Food Additives and Contaminants - Part A Chemistry, Analysis, Control, Exposure and Risk Assessment | Year: 2012

Veterinary drugs, such as antimicrobial compounds, are widely used in poultry and may lead to the presence of residues in matrices of animal origin, such as muscle and liver tissue. In this study, broilers received an experimental feed containing sulfadiazine or doxycycline at cross-contamination levels of 2.5, 5 and 10% of the therapeutic dose in feed. Breast and thigh muscle and liver samples were collected during treatment and depletion period and analysed via liquid chromatography-tandem mass spectrometry (LC-MS/MS). Concentrations reached a plateau phase 3-5 days after the start of experimental feeding. A rapid depletion of residues was noted after withdrawal of the experimental feed. No significant differences in measured concentrations were observed between the various muscle types. Residue concentrations for some experimental groups; the 10% group of sulfadiazine and the 5 and 10% group of doxycycline, however, exceeded their corresponding maximum residue limits (MRLs). © 2012 Taylor & Francis.


Patent
University of Liège, Free University of Colombia and Center Deconomie Rurale | Date: 2010-01-15

It refers to a recombinant alpha-hemolysin polypeptide of Staphylococcus aureus, comprising a deletion in the stem domain, wherein at least one heterologous sequence is inserted in a region selected from the group consisting of regions defined by amino acid position of 108 to 151, 1 to 5, 288 to 293, 43 to 48, 235 to 240, 92 to 97, 31 to 36,or 156 to 161 of SEQ ID NO: 1, with the proviso that, if the heterologous sequence contains five or more consecutive histidines the moiety of the heterologous sequence other than the moiety of five or more consecutive histidines has a minimum length of 11 amino acids; or a variant thereof comprising 1-50 amino acids added, substituted or deleted in SEQ ID NO. 1 and the activity to form oligomers and to bind to lipidic bilayers. It also provides a medicament and vaccine comprising said recombinant polypeptide.


Grant
Agency: European Commission | Branch: FP7 | Program: MC-IAPP | Phase: FP7-PEOPLE-IAPP-2008 | Award Amount: 1.05M | Year: 2009

The ability to increase milk production in cow by bovine somatotropin (BST) was first demonstrated in 1930s. The use of recombinant bovine somatotropin (rBST) in dairy cows has become a common practice in the Untied States (US) and many other countries as the commercial product became available in 1994. However the use and sale of rBST in the EU has never been approved and was banned in 1999 due to concerns on animal health and welfare, food safety and quality, and human health implication associated with the administration of rBST in dairy cows. Nevertheless, there are no direct methods available to date that are capable to detect rBST. The present proposal will, by the use of new technologies and a wide range of expertise, deliver a means of screening and confirming the presence of this unwanted growth promoter in milk. The collaboration within the project will bring together a university, a public institution and two private commercial diagnostic companies to produce novel solutions for monitoring the quality and safety of foods. The proposed research project will give an opportunity of Industry-Academia collaboration that will allow the transfer of high level scientific research into much needed commercial outputs. As a consequence, both academic and industrial partners as well as communities will all benefit, not only during the project but well beyond. The major research outcome will be the delivery of a rapid and simple screening test (dipstick and/or ELISA), a highly accurate and quantitative immuno-biosensor test, and a sensitive and specific chemical confirmatory test for rapid detection and unequivocal identification of the presence of rBST in milk. The formation of partnerships will strengthen the joint efforts to advance research in food safety and quality which benefits ultimately all partners involved, the consumers and the community as a whole.


The present invention refers to a recombinant single-chain alpha-hemolysin polypeptide of Staphylococcus aureus, having a deletion in the stem domain for removing hemolytic activity, wherein at least one heterologous sequence is inserted in a region selected from the group consisting of regions defined by amino acid position 108 to 151, amino acid position 1 to 5, amino acid position 288 to 293, amino acid position 43 to 48, amino acid position 235 to 240, amino acid position 92 to 97, amino acid position 31 to 36, amino acid position 156 to 161 in respect to the wild-type sequence SEQ ID NO: 1, with the proviso that, if the heterologous sequence contains five or more consecutive histidine residues the moiety of the heterologous sequence other than the moiety represented by said five or more consecutive histidine residues has a minimum length of 11 amino acid residues;or a variant thereof, wherein in addition to said deletion in the stem domain and said insertion of heterologous sequence, 1 to 50 amino acid residues are added, substituted or deleted in respect to the wild-type sequence SEQ ID NO: 1 and has the activity to form oligomers and to bind to lipidic bilayers or cell membranes. The present invention also provides a medicament and vaccine comprising said recombinant polypeptide.

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