Allnutt T.R.,Food and Environmental Research Agency |
Ayadi M.,French National Institute for Agricultural Research |
Berben G.,Walloon Agricultural Research Center |
Brodmann P.,Biolytix AG |
Lee D.,UK National Institute of Agricultural Botany
Journal of AOAC International | Year: 2010
Quantification of genetic modification (GM) is often undertaken to test for compliance with the European Union GM labeling threshold in food. Different control laboratories will often use common validated methods, but with different models of real-time PCR machines. We performed two separate ring trials to evaluate the relative precision and accuracy of different types of real-time PCR machines used to quantify the concentration of GM maize. Both trials used dual-labeled fluorogenic probes for quantification. The first ring trial used separate GM and reference assays (a single fluorescence channel), and the second used a combined duplex assay (two simultaneous fluorescence channels). Five manufacturers and seven models-including a 96-well microtiter-plate, rotary, and portable machines-were examined. In one trial, the machine used had a significant effect on precision, but in the other it did not. Overall, the degree of variation due to the machine model was lower than other factors. No significant repeatable difference in accuracy was observed between machine models. It was not possible to use sufficient replication of machine type in each laboratory to examine all sources of variation in this study, but the results strongly indicate that factors other than machine type or manufacturer (e.g., method or laboratory) contribute more to variation in a GM quantification result.
PubMed | Ghent University, Silsoe Spray Applications Unit, Catholic University of the Sacred Heart, Northumbria University and 3 more.
Type: | Journal: The Science of the total environment | Year: 2014
The EU Directive on the sustainable use of pesticides (EU128/2009/EC) requires European Member States to develop training activities targeting occupational exposure to pesticides, and communication material aimed at residents and bystanders. Risk perceptions, knowledge and attitudes associated with passive and occupational exposure to pesticide potentially influence the extent to which different stakeholders adopt self-protective behaviour. A methodology for assessing the link between attitudes, adoption of self-protective behaviours and exposure was developed and tested. A survey was implemented in the Greece, Italy and the UK, and targeted stakeholders associated with pesticide exposure linked to orchards, greenhouse crops and arable crops respectively. The results indicated that the adoption of protective measures is low for residents and bystanders, with the exception of residents in Greece, when compared to operators and workers, who tend to follow recommended safety practices. A regression analysis was used to examine the factors affecting the probability of adopting protective measures as well the as the level of exposure in the case of operators and workers where data are available. The results indicate that the likelihood of engaging in self-protective behaviour is not significantly affected by perceptions of own health being affected by pesticides for residents and bystanders. However, operators who perceive that their heath has been negatively affected by the use of pesticides are found to be more likely to adopt self-protective behaviours. Gender and country differences, in perceptions, attitudes and self-protection are also observed. Recommendations for improved communication, in particular for vulnerable groups, are provided.
Hird S.J.,Food and Environmental Research Agency |
Lau B.P.-Y.,Food Research Division |
Schuhmacher R.,University of Natural Resources and Life Sciences, Vienna |
Krska R.,University of Natural Resources and Life Sciences, Vienna
TrAC - Trends in Analytical Chemistry | Year: 2014
As a result of the range and the variety of toxic and undesirable substances in food, which pose a potential hazard to human health, there is an ever-increasing demand for analytical methods that can reliably detect and quantify contaminants and residues in foods. This review presents the state-of-the-art technology used in the determination of trace residues and contaminants in food by liquid chromatography-mass spectrometry (LC-MS). LC-MS instruments utilize many different types of mass analyzer to improve selectivity and also confidence in assigning the identity of the contaminants detected and to offer different approaches to analysis. We discuss current analytical approaches together with the major benefits and the limitations of these technologies with respect to screening, quantification and identification of contaminants and residues in food. © 2014.
Oplatowska M.,Queen's University of Belfast |
Connolly L.,Queen's University of Belfast |
Stevenson P.,Queen's University of Belfast |
Stead S.,Food and Environmental Research Agency |
Elliott C.T.,Queen's University of Belfast
Analytica Chimica Acta | Year: 2011
Malachite Green (MG), Crystal Violet (CV) and Brilliant Green (BG) are antibacterial, antifungal and antiparasitic agents that have been used for treatment and prevention of diseases in fish. These dyes are metabolized into reduced leuco forms (LMG, LCV, LBG) that can be present in fish muscles for a long period. Due to the carcinogenic properties they are banned for use in fish for human consumption in many countries including the European Union and the United States. HPLC and LC-MS techniques are generally used for the detection of these compounds and their metabolites in fish. This study presents the development of a fast enzyme-linked immunosorbent assay (ELISA) method as an alternative for screening purposes. A first monoclonal cell line producing antibodies to MG was generated using a hybridoma technique. The antibody had good cross-reactivates with related chromatic forms of triphenylmethane dyes such as CV, BG, Methyl Green, Methyl Violet and Victoria Blue R. The monoclonal antibody (mAb) was used to develop a fast (20min) disequilibrium ELISA screening method for the detection of triphenylmethanes in fish. By introducing an oxidation step with 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) during sample extraction the assay was also used to detect the presence of the reduced metabolites of triphenylmethanes. The detection capability of the assay was 1ngg-1 for MG, LMG, CV, LCV and BG which was below the minimum required performance limit (MRPL) for the detection method of total MG (sum of MG and LMG) set by the Commission Decision 2004/25/EC (2ngg-1). The mean recoveries for fish samples spiked at 0.5 MRPL and MRPL levels with MG and LMG were between 74.9 and 117.0% and inter- and intra-assay coefficients of variation between 4.7 and 25.7%. The validated method allows the analysis of a batch of 20 samples in two to three hours. Additionally, this procedure is substantially faster than other ELISA methods developed for MG/LMG thus far. The stable and efficient monoclonal cell line obtained is an unlimited source of sensitive and specific antibody to MG and other triphenylmethanes. © 2011 Elsevier B.V.
Derbyshire N.,University of Leeds |
White S.J.,University of Leeds |
Bunka D.H.J.,University of Leeds |
Song L.,University of Leeds |
And 5 more authors.
Analytical Chemistry | Year: 2012
We have used systematic evolution of ligands by exponential enrichment (SELEX) to isolate RNA aptamers against aminoglycoside antibiotics. The SELEX rounds were toggled against four pairs of aminoglycosides with the goal of isolating reagents that recognize conserved structural features. The resulting aptamers bind both of their selection targets with nanomolar affinities. They also bind the less structurally related targets, although they show clear specificity for this class of antibiotics. We show that this lack of aminoglycoside specificity is a common property of aptamers previously selected against single compounds and described as "specific". Broad target specificity aptamers would be ideal for sensors detecting the entire class of aminoglycosides. We have used ligand-induced aggregation of gold-nanoparticles coated with our aptamers as a rapid and sensitive assay for these compounds. In contrast to DNA aptamers, unmodified RNA aptamers cannot be used as the recognition ligand in this assay, whereas 2′-fluoro-pyrimidine derivatives work reliably. We discuss the possible application of these reagents as sensors for drug residues and the challenges for understanding the structural basis of aminoglycoside-aptamer recognition highlighted by the SELEX results. © 2012 American Chemical Society.
Ferguson C.T.J.,University of Leeds |
O'Neill T.L.,University of Leeds |
Audsley N.,Food and Environmental Research Agency |
Isaac R.E.,University of Leeds
Journal of Experimental Biology | Year: 2015
The polyphagous Drosophila suzukii is a highly invasive species that causes extensive damage to a wide range of berry and stone fruit crops. A better understanding of its biology and especially its behaviour will aid the development of new control strategies. We investigated the locomotor behaviour of D. suzukii in a semi-natural environment resembling a typical summer in northern England and show that adult female D. suzukii are at least 4-fold more active during daylight hours than adult males. This result was reproduced in several laboratory environments and was shown to be a robust feature of mated, but not virgin, female flies. Both males and virgin females kept on a 12 h light:12 h dark (12LD) cycle and constant temperature displayed night-time inactivity (sleep) followed by weak activity in the morning, an afternoon period of quiescence (siesta) and then a prominent evening peak of activity. Both the siesta and the sharp evening peak at lights off were severely reduced in females after mating. Flies of either sex entrained in 12LD displayed a circadian pattern of activity in constant darkness confirming the importance of an endogenous clock in regulating adult activity. This response of females to mating is similar to that elicited in female Drosophila melanogaster by the male sex peptide (SP). We used mass spectrometry to identify a molecular ion (m/z, 5145) corresponding to the poly-hydroxylated SP of D. suzukii and to show that this molecule is transferred to the female reproductive tract during copulation. We propose that the siesta experienced by male and virgin female D. suzukii is an adaptation to avoid unnecessary exposure to the afternoon sun, but that mated females faced with the challenge of obtaining resources for egg production and finding oviposition sites take greater risks, and we suggest that the change in female behaviour is induced by the male SP. © 2015. Published by The Company of Biologists Ltd.
Audsley N.,Food and Environmental Research Agency |
Down R.E.,Food and Environmental Research Agency |
Isaac R.E.,University of Leeds
Peptides | Year: 2015
Drosophila suzukii is a highly polyphagous invasive pest which has been recently introduced into Europe and North America, where it is causing severe economic losses through larval infestations of stone and berry fruits. The peptidome of the selected nervous tissues of adult D. suzukii was investigated as a first step in identifying potential targets for the development of novel insecticides. Through in silico analyses of the D. suzukii genome databases 28 neuropeptide families, comprising more than 70 predicted peptides were identified. Using a combination of liquid chromatography and mass spectrometry of tissue extracts, 33 predicted peptides, representing 15 different peptide families were identified by their molecular masses and a total of 17 peptide sequences were confirmed by ion fragmentation. A comparison between the peptides and precursors of D. suzukii and D. melanogaster shows they are highly conserved, with differences only identified in the amino acid sequences of the peptides encoded in the FMRFamide, hugin and ecydysis triggering hormone precursors. All other peptides predicted and identified from D. suzukii appear to be identical to those previously characterized from D. melanogaster. Adipokinetic hormone was only identified in the corpus cardiacum, other peptides present included short neuropeptide F, a pyrokinin and myosuppressin, the latter of which was the only peptide identified from the crop nerve bundle. Peptides present in extracts of the brain and/or thoracico-abdominal ganglion included allatostatins, cardioacceleratory peptide 2b, corazonin, extended FMRFamides, pyrokinins, myoinihibitory peptides, neuropeptide-like precursor 1, SIFamide, short neuropeptide F, kinin, sulfakinins and tachykinin related peptides. © 2014 Published by Elsevier Inc. All rights reserved.
Isaac R.E.,University of Leeds |
Kim Y.-J.,Gwangju Institute of Science and Technology |
Audsley N.,Food and Environmental Research Agency
Peptides | Year: 2014
The male sex peptide (SP) of Drosophila melanogaster has wide ranging effects on females, including rejection of courting males, increased egg production, changes to the feeding habit, increased synthesis of antimicrobial peptides and elevated locomotor activity during day-time. The peptide activates receptors in sensory neurons of the female reproductive tract and can also traverse into the hemolymph and reach the central nervous system. The SP receptor involved in rejection and egg-laying responses has been shown to be identical to the receptor for the evolutionary conserved myoinhibitory peptides (MIPs) that function as neuropeptides in both males and females. Intriguingly, MIPs cannot substitute for SP when either expressed in the male accessory glands or injected into virgin females. MIPs are linear peptides with an amidated C-terminus which protects them from cleavage by carboxypeptidases, but leaves them exposed to potential attack from aminopeptidase and endopeptidase activities. In contrast, the SP region responsible for eliciting the post-mating response is cyclic and has several hydroxyproline residues N-terminal to the disulfide bridge which is expected to protect the biological activity of SP from peptidases of the male accessory gland and seminal fluid. We now present in vitro data showing that SP is metabolically stable, whereas MIPs are much more susceptible to degradation by peptidases of the male accessory gland and the hemolymph of virgin female D. melanogaster. SP has evolved relatively recently as a MIP receptor ligand that is particularly well adapted to surviving in the hostile degradome of the male accessory gland and seminal fluid. © 2014 Elsevier Inc.
Agatz A.,University of York |
Agatz A.,Food and Environmental Research Agency |
Hammers-Wirtz M.,Research Institute for Ecosystem Analysis and Assessment gaiac |
Gabsi F.,RWTH Aachen |
And 3 more authors.
Environmental Toxicology and Chemistry | Year: 2012
Environmental risk assessment of chemicals is based on single species tests at the individual level with single compounds. However, the protection goal is the sustainability of a population, which faces several natural stressors and mixtures of chemicals in the environment. Therefore, experiments were undertaken to quantify the combined effects of chemicals with different modes of action on Daphnia magna populations. Populations continuously exposed to dispersogen A and at abundance equilibrium were treated with a 2-d pulse of p353-nonylphenol. In previous studies, dispersogen A was shown to act as a natural info-chemical, promoting the reproduction of daphnids (higher offspring quantity) coupled with reduced offspring fitness, whereas nonylphenol in pulsed-exposure caused size-selective mortality. Dispersogen A caused accelerated population growth to maximum abundance, shifted the population structure towards smaller individuals, and increased the population sensitivity to nonylphenol. The authors showed that a positive effect observed at the individual level can be transposed to a negative effect when monitored at the population level. So far, positive effects are not addressed in environmental risk assessment, and even in higher-tier testing, population structure is not quantified. Both factors indicate a potential mismatch between protection aim and risk assessment practice. © 2012 SETAC.
Craig B.H.,University of Edinburgh |
Craig B.H.,Food and Environmental Research Agency |
Pilkington J.G.,University of Edinburgh |
Pemberton J.M.,University of Edinburgh
Journal of Helminthology | Year: 2010
Teladorsagia circumcincta is a polygamous nematode that exhibits morphological polymorphism. Sex ratio is typically female biased and the male nematodes occur in association with the genetically similar, minor morphotypes Teladorsagia davtiani and Teladorsagia trifurcata. In experimental infections, sex ratio (proportion male) and the proportion of minor male morphs observed have been shown to be influenced by both host and nematode-related factors. As similar investigations from natural systems are rare, this study examined whether sex ratio and minor male morph frequency were associated with host age and sex and nematode infra-population size in the isolated Soay sheep population on St Kilda. Count data for Teladorsagia were analysed for sheep of all age classes and both sexes from the winters of three consecutive population crashes (1999, 2002 and 2005). Generally, the intensity of Teladorsagia nematodes increased with host age until the age of 2 years before decreasing. In 2005, abundance of nematodes was generally higher than in the previous crashes, nematode sex ratio was negatively associated with host age and tended to be negatively associated with nematode intensity. Within the male nematode subpopulation, T. circumcincta always predominated, followed by T. davtiani and then T. trifurcata, with little variation in the relative proportions between hosts. The presence of each minor morph was primarily associated with the intensity of male T. circumcincta and, in those hosts where all three male morphs were detected, intensity of each minor morph was most associated with intensity of Teladorsagia females. Therefore, in a year when the nematode was generally more abundant, sex ratio appeared to be influenced by both host and nematode-related factors, whereas in all years examined, the frequency of morphological polymorphism was primarily density dependent. © Cambridge University Press 2009.