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Food consumption play a crucial role in human life, yet conventional food production and consumption patterns can be detrimental to the environment. Thus, research and development has been directed towards alternative proteins, with edible insects being promising sources. Edible insects have been recognised for their sustainable benefits providing protein, with less emission of greenhouse gas, land and water usage compared to sources, such as beef, chicken, and dairy products. Among the over 2000 known edible insect species, only four, namely yellow mealworm (Tenebrio molitor), migratory locust/grasshopper (Locusta migratoria), grain mould beetle, also known as lesser mealworm which is a larval form of Alphitobius diaperinus (from the family of Tenebrionidae of darkling beetles) and house cricket (Acheta domesticus), are currently authorised in specific products through specific producers in the EU. The expansion of such foods into Western diets face challenges such as consumer barriers, gaps in microbiological and chemical safety hazard data during production and processing, and the potential for fraudulent supply chain activity. The main aim of this study was to map the supply chain, through interviews with personnel along the supply chain, coupled with searches for relevant publications and governmental documents. Thus, the main potential points of food safety and fraud along the edible insect supply chain were identified. Feed substrate was identified as the main area of concern regarding microbiological and chemical food safety and novel processing techniques were forecast to be of most concern for future fraudulent activity. Despite the on-going authorisation of insect species in many countries there are substantial food safety and authenticity information gaps in this industry that need to be addressed before edible insects can be viewed as a safe and sustainable protein sources by Western consumers.
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Smartphone based devices (SBDs) have the potential to revolutionize food safety control by empowering citizens to perform screening tests. To achieve this, it is of paramount importance to understand current research efforts and identify key technology gaps. Therefore, a systematic review of optical SBDs in the food safety sector was performed. An overview of reviewed SBDs is given focusing on performance characteristics as well as image analysis procedures. The state-of-the-art on commercially available SBDs is also provided. This analysis revealed several important technology gaps, the most prominent of which are: (i) the need to reach a consensus regarding optimal image analysis, (ii) the need to assess the effect of measurement variation caused by using different smartphones and (iii) the need to standardize validation procedures to obtain robust data. Addressing these issues will drive the development of SBDs and potentially unlock their massive potential for citizen-based food control.
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Detailed knowledge regarding sensor based technologies for the detection of food contamination often remains concealed within scientific journals or divided between numerous commercial kits which prevents optimal connectivity between companies and end-users. To overcome this barrier The End user Sensor Tree (TEST) has been developed. TEST is a comprehensive, interactive platform including over 900 sensor based methods, retrieved from the scientific literature and commercial market, for aquatic-toxins, mycotoxins, pesticides and microorganism detection. Key analytical parameters are recorded in excel files while a novel classification system is used which provides, tailor-made, experts' feedback using an online decision tree and database introduced here. Additionally, a critical comparison of reviewed sensors is presented alongside a global perspective on research pioneers and commercially available products. The lack of commercial uptake of the academically popular electrochemical and nanomaterial based sensors, as well as multiplexing platforms became very apparent and reasons for this anomaly are discussed.
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Técnicas Biosensibles/métodos , Contaminación de Alimentos/análisis , Micotoxinas/aislamiento & purificación , Plaguicidas/aislamiento & purificación , Técnicas Biosensibles/clasificación , Bases de Datos Factuales , Humanos , Micotoxinas/química , Nanoestructuras/química , Plaguicidas/químicaRESUMEN
Research in biosensing approaches as alternative techniques for food diagnostics for the detection of chemical contaminants and foodborne pathogens has increased over the last twenty years. The key component of such tests is the biorecognition element whereby polyclonal or monoclonal antibodies still dominate the market. Traditionally the screening of sera or cell culture media for the selection of polyclonal or monoclonal candidate antibodies respectively has been performed by enzyme immunoassays. For niche toxin compounds, enzyme immunoassays can be expensive and/or prohibitive methodologies for antibody production due to limitations in toxin supply for conjugate production. Automated, self-regenerating, chip-based biosensors proven in food diagnostics may be utilised as rapid screening tools for antibody candidate selection. This work describes the use of both single channel and multi-channel surface plasmon resonance (SPR) biosensors for the selection and characterisation of antibodies, and their evaluation in shellfish tissue as standard techniques for the detection of domoic acid, as a model toxin compound. The key advantages in the use of these biosensor techniques for screening hybridomas in monoclonal antibody production were the real time observation of molecular interaction and rapid turnaround time in analysis compared to enzyme immunoassays. The multichannel prototype instrument was superior with 96 analyses completed in 2h compared to 12h for the single channel and over 24h for the ELISA immunoassay. Antibodies of high sensitivity, IC50's ranging from 4.8 to 6.9ng/mL for monoclonal and 2.3-6.0ng/mL for polyclonal, for the detection of domoic acid in a 1min analysis time were selected. Although there is a progression for biosensor technology towards low cost, multiplexed portable diagnostics for the food industry, there remains a place for laboratory-based SPR instrumentation for antibody development for food diagnostics as shown herein.
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Análisis de los Alimentos/métodos , Ácido Kaínico/análogos & derivados , Toxinas Marinas/análisis , Mariscos/análisis , Resonancia por Plasmón de Superficie/métodos , Animales , Anticuerpos Inmovilizados/química , Anticuerpos Monoclonales/química , Diseño de Equipo , Análisis de los Alimentos/instrumentación , Hibridomas , Inmunoensayo/instrumentación , Inmunoensayo/métodos , Ácido Kaínico/análisis , Límite de Detección , Ratones Endogámicos BALB C , Conejos , Resonancia por Plasmón de Superficie/instrumentaciónRESUMEN
The aim of the study was to investigate the potential of a metabolomics platform to distinguish between pigs treated with ronidazole, dimetridazole and metronidazole and non-medicated animals (controls), at two withdrawal periods (day 0 and 5). Livers from each animal were biochemically profiled using UHPLC-QTof-MS in ESI+ mode of acquisition. Several Orthogonal Partial Least Squares-Discriminant Analysis models were generated from the acquired mass spectrometry data. The models classified the two groups control and treated animals. A total of 42 ions of interest explained the variation in ESI+. It was possible to find the identity of 3 of the ions and to positively classify 4 of the ionic features, which can be used as potential biomarkers of illicit 5-nitroimidazole abuse. Further evidence of the toxic mechanisms of 5-nitroimidazole drugs has been revealed, which may be of substantial importance as metronidazole is widely used in human medicine.
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Biomarcadores/análisis , Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos/metabolismo , Metabolómica/métodos , Nitroimidazoles/efectos adversos , Animales , Espectrometría de Masas/métodos , PorcinosRESUMEN
Food allergy is an increasing problem for those affected, their families or carers, the food industry and for regulators. The food supply chain is highly vulnerable to fraud involving food allergens, risking fatalities and severe reputational damage to the food industry. Many facets are being pursued to ameliorate the difficulties including better food labelling and the concept of thresholds of elicitation of allergy symptoms as risk management tools. These efforts depend to a high degree on the ability reliably to detect and quantify food allergens; yet all current analytical approaches exhibit severe deficiencies that jeopardise accurate results being produced particularly in terms of the risks of false positive and false negative reporting. If we fail to realise the promise of current risk assessment and risk management of food allergens through lack of the ability to measure food allergens reproducibly and with traceability to an international unit of measurement, the analytical community will have failed a significant societal challenge. Three distinct but interrelated areas of analytical work are urgently needed to address the substantial gaps identified: (a) a coordinated international programme for the production of properly characterised clinically relevant reference materials and calibrants for food allergen analysis; (b) an international programme to widen the scope of proteomics and genomics bioinformatics for the genera containing the major allergens to address problems in ELISA, MS and DNA methods; (c) the initiation of a coordinated international programme leading to reference methods for allergen proteins that provide results traceable to the SI. This article describes in more detail food allergy, the risks of inapplicable or flawed allergen analyses with examples and a proposed framework, including clinically relevant incurred allergen concentrations, to address the currently unmet and urgently required analytical requirements. Support for the above recommendations from food authorities, business organisations and National Measurement Institutes is important; however transparent international coordination is essential. Thus our recommendations are primarily addressed to the European Commission, the Health and Food Safety Directorate, DG Santé. A global multidisciplinary consortium is required to provide a curated suite of data including genomic and proteomic data on key allergenic food sources, made publically available on line.
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Alérgenos/análisis , Análisis de los Alimentos/métodos , Abastecimiento de Alimentos , Salud , Animales , Humanos , RiesgoRESUMEN
Mycotoxins and heavy metals are ubiquitous in the environment and contaminate many foods. The widespread use of pesticides in crop production to control disease contributes further to the chemical contamination of foods. Thus multiple chemical contaminants threaten the safety of many food commodities; hence the present study used maize as a model crop to identify the severity in terms of human exposure when multiple contaminants are present. High Content Analysis (HCA) measuring multiple endpoints was used to determine cytotoxicity of complex mixtures of mycotoxins, heavy metals and pesticides. Endpoints included nuclear intensity (NI), nuclear area (NA), plasma membrane permeability (PMP), mitochondrial membrane potential (MMP) and mitochondrial mass (MM). At concentrations representing legal limits of each individual contaminant in maize (3ng/ml ochratoxin A (OTA), 1µg/ml fumonisin B1 (FB1), 2ng/ml aflatoxin B1 (AFB1), 100ng/ml cadmium (Cd), 150ng/ml arsenic (As), 50ng/ml chlorpyrifos (CP) and 5µg/ml pirimiphos methyl (PM), the mixtures (tertiary mycotoxins plus Cd/As) and (tertiary mycotoxins plus Cd/As/CP/PM) were cytotoxic for NA and MM endpoints with a difference of up to 13.6% (p≤0.0001) and 12% (p≤0.0001) respectively from control values. The most cytotoxic mixture was (tertiary mycotoxins plus Cd/As/CP/PM) across all 4 endpoints (NA, NI, MM and MMP) with increases up to 61.3%, 23.0%, 61.4% and 36.3% (p≤0.0001) respectively. Synergy was evident for two endpoints (NI and MM) at concentrations contaminating maize above legal limits, with differences between expected and measured values of (6.2-12.4% (p≤0.05-p≤0.001) and 4.5-12.3% (p≤0.05-p≤0.001) for NI and MM, respectively. The study introduces for the first time, a holistic approach to identify the impact in terms of toxicity to humans when multiple chemical contaminants are present in foodstuffs. Governmental regulatory bodies must begin to contemplate how to safeguard the population when such mixtures of contaminants are found in foods and this study starts to address this critical issue.
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Mezclas Complejas/toxicidad , Contaminación de Alimentos/análisis , Metales Pesados/toxicidad , Micotoxinas/toxicidad , Plaguicidas/toxicidad , Zea mays/química , Animales , Permeabilidad de la Membrana Celular/efectos de los fármacos , Núcleo Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Perros , Relación Dosis-Respuesta a Droga , Humanos , Células de Riñón Canino Madin Darby , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Metales Pesados/aislamiento & purificación , Micotoxinas/aislamiento & purificación , Plaguicidas/aislamiento & purificación , Medición de RiesgoRESUMEN
Honey is a high value food commodity with recognized nutraceutical properties. A primary driver of the value of honey is its floral origin. The feasibility of applying multivariate data analysis to various chemical parameters for the discrimination of honeys was explored. This approach was applied to four authentic honeys with different floral origins (rata, kamahi, clover and manuka) obtained from producers in New Zealand. Results from elemental profiling, stable isotope analysis, metabolomics (UPLC-QToF MS), and NIR, FT-IR, and Raman spectroscopic fingerprinting were analyzed. Orthogonal partial least square discriminant analysis (OPLS-DA) was used to determine which technique or combination of techniques provided the best classification and prediction abilities. Good prediction values were achieved using metabolite data (for all four honeys, Q(2)=0.52; for manuka and clover, Q(2)=0.76) and the trace element/isotopic data (for manuka and clover, Q(2)=0.65), while the other chemical parameters showed promise when combined (for manuka and clover, Q(2)=0.43).
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Fenómenos Químicos , Miel/análisis , Bases de Datos Factuales , Análisis Discriminante , Flores/química , Análisis de los Alimentos , Metabolómica , Nueva Zelanda , Reproducibilidad de los Resultados , Espectroscopía Infrarroja por Transformada de Fourier , Espectrometría RamanRESUMEN
Azaspiracid (AZA) poisoning was unknown until 1995 when shellfish harvested in Ireland caused illness manifesting by vomiting and diarrhoea. Further in vivo/vitro studies showed neurotoxicity linked with AZA exposure. However, the biological target of the toxin which will help explain such potent neurological activity is still unknown. A region of Irish coastline was selected and shellfish were sampled and tested for AZA using mass spectrometry. An outbreak was identified in 2010 and samples collected before and after the contamination episode were compared for their metabolite profile using high resolution mass spectrometry. Twenty eight ions were identified at higher concentration in the contaminated samples. Stringent bioinformatic analysis revealed putative identifications for seven compounds including, glutarylcarnitine, a glutaric acid metabolite. Glutaric acid, the parent compound linked with human neurological manifestations was subjected to toxicological investigations but was found to have no specific effect on the sodium channel (as was the case with AZA). However in combination, glutaric acid (1 mM) and azaspiracid (50 nM) inhibited the activity of the sodium channel by over 50%. Glutaric acid was subsequently detected in all shellfish employed in the study. For the first time a viable mechanism for how AZA manifests itself as a toxin is presented.
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Enfermedades Transmitidas por los Alimentos/etiología , Toxinas Marinas/química , Toxinas Marinas/toxicidad , Mariscos/análisis , Mariscos/toxicidad , Compuestos de Espiro/química , Compuestos de Espiro/toxicidad , Animales , Bivalvos/anatomía & histología , Bivalvos/química , Carnitina/análogos & derivados , Carnitina/química , Línea Celular , Línea Celular Tumoral , Brotes de Enfermedades , Glutaratos/química , Células HEK293 , Humanos , Canales de Sodio/metabolismoRESUMEN
Aflatoxin B1 (AFB1), ochratoxin A (OTA) and fumonisin B1 (FB1) are important mycotoxins in terms of human exposure via food, their toxicity and regulatory limits that exist worldwide. Mixtures of toxins can frequently be present in foods, however due to the complications of determining their combined toxicity, legal limits of exposure are determined for single compounds, based on long standing toxicological techniques. High content analysis (HCA) may be a useful tool to determine total toxicity of complex mixtures of mycotoxins. Endpoints including cell number (CN), nuclear intensity (NI), nuclear area (NA), plasma membrane permeability (PMP), mitochondrial membrane potential (MMP) and mitochondrial mass (MM) were compared to the conventional 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) and neutral red (NR) endpoints in MDBK cells. Individual concentrations of each mycotoxin (OTA 3 µg/ml, FB1 8 µg/ml and AFB1 1.28 µg/ml) revealed no cytotoxicity with MTT or NR but HCA showed significant cytotoxic effects up to 41.6% (p≤0.001) and 10.1% (p≤0.05) for OTA and AFB1, respectively. The tertiary mixture (OTA 3 µg/ml, FB1 8 µg/ml and AFB1 1.28 µg/ml) detected up to 37.3% and 49.8% more cytotoxicity using HCA over MTT and NR, respectively. Whilst binary combinations of OTA (3 µg/ml) and FB1 (8 µg/ml) revealed synergistic interactions using HCA (MMP, MM, NI endpoints) not detected using MTT or NR. HCA is a highly novel and sensitive tool that could substantially help determine future regulatory limits, for single and combined toxins present in food, ensuring legislation is based on true risks to human health exposure.
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Aflatoxina B1/toxicidad , Contaminación de Alimentos , Fumonisinas/toxicidad , Ocratoxinas/toxicidad , Animales , Bovinos , Permeabilidad de la Membrana Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Sinergismo Farmacológico , Potencial de la Membrana Mitocondrial/efectos de los fármacosRESUMEN
Industrial chemicals, antimicrobials, drugs and personal care products have been reported as global pollutants which enter the food chain. Some of them have also been classified as endocrine disruptors based on results of various studies employing a number of in vitro/vivo tests. The present study employed a mammalian reporter gene assay to assess the effects of known and emerging contaminants on estrogen nuclear receptor transactivation. Out of fifty-nine compounds assessed, estrogen receptor agonistic activity was observed for parabens( n = 3), UV filters (n = 6), phthalates (n = 4) and a metabolite, pyrethroids (n = 9) and their metabolites (n = 3). Two compounds were estrogen receptor antagonists while some of the agonists enhanced 17b-estradiol mediated response.This study reports five new compounds (pyrethroids and their metabolites) possessing estrogen agonist activity and highlights for the first time that pyrethroid metabolites are of particular concern showing much greater estrogenic activity than their parent compounds.
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Disruptores Endocrinos/toxicidad , Cadena Alimentaria , Contaminación de Alimentos , Línea Celular , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Genes Reporteros/efectos de los fármacos , Humanos , Parabenos/toxicidad , Ácidos Ftálicos/toxicidad , Piretrinas/toxicidad , Receptores de Estrógenos/antagonistas & inhibidores , Pruebas de ToxicidadRESUMEN
Dietary crude protein (CP) and phosphorus (P) have the potential to alter dairy cow production, nutrient status, and milk heat stability, specifically in early lactation. This study examined the effect of supplementary concentrates with different CP and P concentrations on blood N and P status and on milk yield, composition, and heat stability. The concentrates [4kg of dry matter (DM) concentrate per cow daily] were fed to grazing dairy cows (13kg DM grass) during early lactation. Forty-eight spring-calving dairy cows were allocated to 4 treatments: high CP, high P (HPrHP; 302g/kg DM CP, 6.8g/kg DM P), medium CP, high P (MPrHP; 202g/kg DM CP, 4.7g/kg DM P), low CP, high P (LPrHP; 101g/kg DM CP, 5.1g/kg DM P), and low CP, low P (LPrLP; 101g/kg DM CP, 0.058g/kg DM P), for 8wk. Levels of N excretion were significantly higher in animals fed the HPrHP and MPrHP concentrates; P excretion was significantly lower in animals fed the LPrLP concentrate. Reducing the level of P in the diet (LPrLP concentrate) resulted in a significantly lower blood P concentration, whereas milk yield and composition (fat and protein) were not affected by either CP or P in the diet. The effect of the interaction between treatment and time on milk urea N was significant, reflecting the positive correlation between dietary CP and milk nonprotein N. Increasing supplementary CP and P (HPrHP) in the diet resulted in significantly lower milk heat stability at pH 6.8. The findings show that increasing dietary CP caused a decrease in milk heat stability, which reduced the suitability of milk for processing. The study also found that increasing dietary CP increased milk urea N and milk nonprotein N. Increasing dietary P increased fecal P excretion. These are important considerations for milk processors and producers for control of milk processing and environmental parameters.
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Fenómenos Fisiológicos Nutricionales de los Animales , Bovinos/fisiología , Proteínas en la Dieta/metabolismo , Lactancia/fisiología , Leche , Fósforo Dietético/metabolismo , Animales , Suplementos Dietéticos/análisis , Femenino , Leche/química , Leche/metabolismo , Leche/fisiología , Nitrógeno/metabolismoRESUMEN
Endocrine disruptors (EDs) are compounds known to interfere with the endocrine system by disturbing the action or pathways of natural hormones which may lead to infertility or cancer. Our diet is considered to be one of the main exposure routes to EDs. Since milk and dairy products are major components of our diet they should be monitored for ED contamination. Most assays developed to date utilise targeted, chromatography based methods which lack information on the biological activity and mixture effects of the monitored compounds. A biological reporter gene assay (RGA) was developed to assess the total estrogen hormonal load in milk. It has been validated according to EU decision 2002/657/EC. Analytes were extracted by liquid-liquid extraction with acetonitrile followed by clean up on a HLB column which yielded good recovery and small matrix effects. The method has been shown to be estrogen specific, repeatable and reproducible, with covariance values below 20%. In conclusion, this method enables the detection of low levels of estrogen hormonal activity in milk with a detection capability of 36 pg g(-)(1) EEQ and has been successfully applied in testing a range of milk samples.
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Disruptores Endocrinos/análisis , Contaminación de Alimentos/análisis , Genes Reporteros , Leche/química , Animales , Línea Celular , Disruptores Endocrinos/toxicidad , Humanos , Límite de Detección , Extracción Líquido-Líquido , Luciferasas/genética , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
Spiramycin, tylosin, bacitracin and virginiamycin are among a group of antibiotic growth promoters that have been banned in the European Union since the 1999 Council. This was due to concerns over the development of resistant bacteria emerging between humans and animals with the threat of antibiotics no longer being able to be used effectively to treat human infections. A sensitive and fast immunochemical method is presented for the determination of these four antibiotic growth promoters simultaneously in poultry tissue. The method employs methanol extraction followed by sample clean-up by solid-phase extraction (SPE) with determination by enzyme-linked immunoabsorbant assay (ELISA). The limit of detection (LOD) was less than 1 ng g(-1) and the detection capability (CCß) was 3 ng g(-1) or less for all four antibiotic growth promoters. Validation was completed with both raw and cooked chicken, therefore either matrix could be used for the monitoring of these banned drugs. In a feeding trial no residues of either bacitracin or virginiamycin were found in medicated birds even without a withdrawal period. In the case of tylosin and spiramycin much higher residues level were detected immunochemically than was the case by mass spectrometry.
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Antibacterianos/análisis , Sustancias de Crecimiento/análisis , Productos Avícolas/análisis , Animales , Ensayo de Inmunoadsorción Enzimática , Límite de DetecciónRESUMEN
AIMS: The objective of this study was to develop a novel screening method for detection of viable Mycobacterium avium subsp. paratuberculosis (Map) in milk and faeces, as a rapid alternative to Map culture. METHODS AND RESULTS: The new method couples Map-specific peptide-mediated magnetic separation technique with an optimized phage amplification assay followed by detection of released progeny phage by ELISA in a competition assay format using polyclonal antibody produced against the D29 mycobacteriophage involved in the phage assay. Sample matrices were found not to interfere with the developed method, and the dynamic range of the assay was 3 × 10(2) -6 × 10(8 ) phage ml(-1) . When low numbers of Map were present (10(2) CFU ml(-1) ), the burst size of a single host Map cell was maximal (10(3) phage per cell) resulting in a highly sensitive screening assay. CONCLUSION: A rapid, sensitive immuno-based screening method suitable for the detection of viable Map in milk and faeces was developed. SIGNIFICANCE AND IMPACT OF THE STUDY: The novel PMS-phage-ELISA permits sensitive, qualitative detection of viable Map in milk or faeces samples within 48 h, representing a substantial decrease in time to detection compared with current culture methods for Map.
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Ensayo de Inmunoadsorción Enzimática/métodos , Mycobacterium avium subsp. paratuberculosis/aislamiento & purificación , Animales , Heces/microbiología , Leche/microbiología , Micobacteriófagos/inmunologíaRESUMEN
Low emission slurry spreading techniques are known to improve nitrogen use efficiency, but their impact on phosphorus (P) losses in surface runoff has received little attention. The current study was designed to examine the effect of slurry spreading technique on P losses in runoff. Twelve treatments were examined on 0.5- m by 1.0-m plots in a nominal 2 × 6 factorial design experiment. Treatments comprised grass swards at two different stages of growth, a stubble and a 4-wk regrowth, and six different slurry application treatments: control (no slurry), and slurry applied to simulate splash-plate, injection (across and down slope), and trailing shoe (across and down slope) spreading. Slurry was applied by hand (40 m ha). Rainfall simulations (40 mm h) were conducted at 2, 9, and 28 d post-slurry application. When slurry was applied to the stubble, dissolved reactive P (DRP) concentrations in runoff at Day 2 were 47 and 37% lower ( < 0.05) from the injection and trailing shoe treatments compared with the splash-plate treatment. Similarly, at Day 2, TP concentrations in runoff from the injection treatments were 27% lower ( < 0.05) than the splash-plate treatment. In contrast, application technique had no effect ( 0.05) on P concentrations in runoff following slurry application to the regrowth treatment. Phosphorus concentrations in runoff were unaffected by direction of slurry spreading (across or down) at both applications. Our results indicate that trailing shoe and injection techniques offer the potential to reduce DRP concentrations in runoff during the period immediately after slurry application.
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Estiércol , Fósforo , Nitrógeno , Poaceae , LluviaRESUMEN
AIMS: The objectives of this study were to produce Salmonella-specific peptide ligands by phage display biopanning and evaluate their use for magnetic separation (MS). METHODS AND RESULTS: Four-phage display biopanning rounds were performed, and the peptides expressed by the two most Salmonella-specific (on the basis of phage-binding ELISA results) phage clones, MSal020401 and MSal020417, were chemically synthesized and coupled to MyOne™ tosylactivated Dynabeads(®). Peptide capture capability for whole Salmonella cells from nonenriched broth cultures was quantified by MS + plate counts and MS + Greenlight™ detection and compared to capture capability of anti-Salmonella (antibody-coated) Dynabeads(®). MS + Greenlight™ gave a more comprehensive picture of capture capability than MS + plate counts and showed that Peptide MSal020417-coated beads exhibited at least similar, if not better, capture capability to anti-Salmonella Dynabeads(®) (mean capture values of 36·0 ± 18·2 and 31·2 ± 20·1%, respectively, over Salmonella spp. concentration range 3 × 10(1) -3 × 10(6) CFU ml(-1)) with cross-reactivity of ≤1·9% to three other foodborne pathogens: Escherichia coli, Listeria monocytogenes and Campylobacter jejuni. CONCLUSIONS: One of the phage display-derived peptide ligands was demonstrated by MS + Greenlight™ to be a viable antibody alternative for MS of Salmonella spp. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates an antibody-free approach to Salmonella detection and opens substantial possibilities for more rapid tests for this bacterium.
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Técnicas de Visualización de Superficie Celular , Péptidos/metabolismo , Salmonella/aislamiento & purificación , Ligandos , Fenómenos Magnéticos , Péptidos/químicaRESUMEN
Ochratoxin A (OTA) is a mycotoxin and extrolite of fungi which has been reported in a range of foods. This study uses mammalian reporter gene assays (RGAs) with natural steroid receptors and the H295R steroidogenesis assay to assess the endocrine disrupting activity of OTA. At the receptor level, OTA (within a concentration range of 0.25-2500 ng/ml) did not induce an agonistic response in an oestrogen, androgen, progestagen or glucocorticoid RGA. An antagonistic effect was observed in all of the RGAs at the highest concentration tested (2500 ng/ml). However, while there was no significant cytotoxic effect observed in the MTT (thiazolyl blue tetrazolium bromide) cell viability assay at this concentration, there was a corresponding change in cell morphology which may be related to the resulting antagonistic effect. At the hormone production level, H295R cells were used as a steroidogenesis model and exposed to OTA (within a concentration range of 0.1-1000 ng/ml). Treatment of the cells with 1000 ng/ml OTA increased the production of estradiol (117±14 ng/ml) over 3 times that of the solvent control (36±9 pg/ml). Western blotting confirmed an increase in aromatase protein. Overall the results indicate that OTA does not appear to interact with steroid receptors but has the potential to cause endocrine disruption by interfering with steroidogenesis. This is the first study identifying the effect OTA may have on production of the steroid hormone estradiol.
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Disruptores Endocrinos/toxicidad , Estradiol/biosíntesis , Ocratoxinas/toxicidad , Receptores Androgénicos/metabolismo , Receptores de Estrógenos/metabolismo , Receptores de Glucocorticoides/metabolismo , Receptores de Progesterona/metabolismo , Aromatasa/biosíntesis , Western Blotting , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Genes Reporteros , Humanos , Receptores Androgénicos/genética , Receptores de Estrógenos/genética , Receptores de Glucocorticoides/genética , Receptores de Progesterona/genéticaRESUMEN
Mass spectrometric methods were developed and validated for the analysis in chicken muscle of a range of antibiotic growth promoters: spiramycin, tylosin, virginiamycin and bacitracin, and separately for two marker metabolites of carbadox (quinoxaline-2-carboxylic acid and 1,4-bisdesoxycarbadox), and a marker metabolite of olaquindox (3-methyl-quinoxaline-2-carboxylic acid). The use of these compounds as antibiotic growth promoters has been banned by the European Commission. This study aimed to develop methods to detect their residues in muscle samples as a means of checking for the use of these drugs during the rearing of broiler chickens. When fed growth-promoting doses for 6 days, spiramycin (31.4 µg kg(-1)), tylosin (1.0 µg kg(-1)), QCA (6.5 µg kg(-1)), DCBX (71.2 µg kg(-1)) and MQCA (0.2 µg kg(-1)) could be detected in the muscle 0 days after the withdrawal of fortified feed. Only spiramycin could consistently be detected beyond a withdrawal period of 1 day. All analytes showed stability to a commercial cooking process, therefore raw or cooked muscle could be used for monitoring purposes.
Asunto(s)
Sustancias de Crecimiento/administración & dosificación , Músculo Esquelético/química , Espectrometría de Masa por Ionización de Electrospray/métodos , Detección de Abuso de Sustancias/métodos , Alimentación Animal , Animales , Pollos , Cromatografía de Gases y Espectrometría de MasasRESUMEN
Although interest in crossbreeding within dairy systems has increased, the role of Jersey crossbred cows within high concentrate input systems has received little attention. This experiment was designed to examine the performance of Holstein-Friesian (HF) and Jersey × Holstein-Friesian (J × HF) cows within a high concentrate input total confinement system (CON) and a medium concentrate input grazing system (GRZ). Eighty spring-calving dairy cows were used in a 2 (cow genotype) × 2 (milk production system) factorial design experiment. The experiment commenced when cows calved and encompassed a full lactation. With GRZ, cows were offered diets containing grass silage and concentrates [70:30 dry matter (DM) ratio] until turnout, grazed grass plus 1.0 kg of concentrate/day during a 199-d grazing period, and grass silage and concentrates (75:25 DM ratio) following rehousing and until drying-off. With CON, cows were confined throughout the lactation and offered diets containing grass silage and concentrates (DM ratio; 40:60, 50:50, 40:40, and 75:25 during d 1 to 100, 101 to 200, 201 to 250, and 251 until drying-off, respectively). Full-lactation concentrate DM intakes were 791 and 2,905 kg/cow for systems GRZ and CON, respectively. Although HF cows had a higher lactation milk yield than J × HF cows, the latter produced milk with a higher fat and protein content, so that solids-corrected milk yield (SCM) was unaffected by genotype. Somatic cell score was higher with the J × HF cows. Throughout lactation, HF cows were on average 37 kg heavier than J × HF cows, whereas the J × HF cows had a higher body condition score. Within each system, food intake did not differ between genotypes, whereas full-lactation yields of milk, fat plus protein, and SCM were higher with CON than with GRZ. A significant genotype × environment interaction was observed for milk yield, and a trend was found for an interaction with SCM. Crossbred cows on CON gained more body condition than HF cows, and overall pregnancy rate was unaffected by either genotype or management system. In summary, milk and SCM yields were higher with CON than with GRZ, whereas genotype had no effect on SCM. However, HF cows exhibited a greater milk yield response and a trend toward a greater SCM yield response with increasing concentrate levels compared with the crossbred cows.