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Fitzgerald J.,Dublin City University | Leonard P.,Dublin City University | Darcy E.,Dublin City University | Danaher M.,Ashtown Food Research Center | O'Kennedy R.,Dublin City University
Analytical Biochemistry | Year: 2011

Halofuginone is an antiprotozoal drug used in the treatment of coccidiosis in poultry, a contagious enteric disease caused by parasites of the Eimeria spp. To ensure that food is free from any halofuginone residues and safe for human consumption, a rapid method to detect these residues below the maximum residue limits (MRLs) in a variety of matrices is necessary. To address this need, we constructed an immune single-chain variable fragment (scFv) library from the RNA of a halofuginone-immunized chicken and selected halofuginone-specific scFv by phage display. The best clone isolated from the library had a limit of detection of 30 ng/ml as determined by enzyme-linked immunosorbent assay (ELISA). However, the minimum MRL for halofuginone in certain foodstuffs can be as low as 1 ng/ml, well below the sensitivity of the selected antibody. The selected antibody was then affinity maturated by light-chain shuffling to further improve the antibody's assay performance. The halofuginone-specific heavy-chain pool of the biopanned library was assembled with the light-chain repertoire amplified from the original prepanned library. This resulted in a heavy-chain-biased library from which an scFv with the potential to detect halofuginone residues as low as 80 pg/ml was isolated, a 185-fold improvement over the original scFv. This new chain-shuffled scFv was incorporated into a validated ELISA (according to Commission Regulation 2002/657/EC) for the sensitive detection of halofuginone in spiked processed egg samples. © 2010 Elsevier Inc. All rights reserved. Source


Fitzgerald J.,Dublin City University | Leonard P.,Dublin City University | Danaher M.,Ashtown Food Research Center | O'Kennedy R.,Dublin City University
Applied Biochemistry and Biotechnology | Year: 2015

This research describes the development of a multi-analyte lateral-flow immunoassay intended for the simultaneous detection of three anti-protozoan drugs (coccidiostats). These drugs, namely, halofuginone, toltrazuril and diclazuril, are used in the treatment of Eimeria spp. infections in cattle, pigs, chickens and turkeys. Coloured carboxylated microspheres were coated with each of the detection antibodies and employed in a lateral-flow assay format for detection of these residues in eggs. Using this approach, halofuginone was detectable at a limit of 10 ng/mL or greater, toltrazuril at 100 ng/mL and, similarly, diclazuril had a detection limit of 100 ng/mL, which is below the maximum allowed residue limit for all three as outlined by EU regulation. This simple cost-efficient assay and analysis method could pave the way for more efficient simultaneous monitoring of small-molecule residues in the future. © 2015, Springer Science+Business Media New York. Source


Mooney A.,Dublin City University | Corry A.J.,Dublin City University | Ruairc C.N.,Dublin City University | Mahgoub T.,Dublin City University | And 8 more authors.
Dalton Transactions | Year: 2010

A series of N-(ferrocenyl)naphthoyl amino acid esters 5-18 has been prepared by coupling ferrocenyl naphthoic acids 3-4 to α-amino acids and linear amino acids in the presence of N-(3-dimethylaminopropyl)-N′- ethylcarbodiimide hydrochloride (EDC) and 1-hydroxybenzotriazole (HOBt). The compounds were fully characterised by a range of NMR spectroscopic techniques, UV-Vis spectroscopy, mass spectrometry and cyclic voltammetry. X-ray crystallographic studies of the intermediate compounds 1-2 were also performed. Biological evaluation of the intermediates 1-2 and N-(ferrocenyl)naphthoyl amino acid esters 5-18 was performed in the H1299 non-small cell lung cancer (NSCLC) cell line and the Sk-Mel-28 metastatic melanoma cell line. The intermediates 1-2 failed to produce an effect in either cell line. Compounds 5-18 exhibited a strong anti-proliferative effect in the H1299 cell line, whilst the Sk-Mel-28 cells were slightly more resistant to these compounds. N-(6-ferrocenyl-2- naphthoyl)-γ-aminobutyric acid ethyl ester 17 shows a particularly high activity in both the H1299 cell line (IC50 = 0.62 ± 0.07 μM) and the Sk-Mel-28 cell line (IC50 = 1.41 ± 0.04 μM). © 2010 The Royal Society of Chemistry. Source


Wasilewski P.D.,University of Technology and Life Sciences in Bydgoszcz | Nowachowicz J.,University of Technology and Life Sciences in Bydgoszcz | Michalska G.,University of Technology and Life Sciences in Bydgoszcz | Bucek T.,University of Technology and Life Sciences in Bydgoszcz | And 2 more authors.
Archiv fur Tierzucht | Year: 2011

The aim of the paper was to investigate the impact of feeding pigs with different levels of conjugated linoleic acid or sunflower oil on fatty acid profile of Longissimus dorsi muscle. The subjects of research were 60 crossbred gilts divided into 6 groups, fed with different levels of conjugated linoleic acid (CLA) or sunflower oil (SFO) (0.5; 1.0 and 2.0 %, respectively). All fatteners were kept and fed under standardized conditions. Animals were slaughtered at 95 kg of body weight. Fatty acid profile was determined in samples of Longissimus dorsi muscle from each animal. Gas chromatography was used (in the research). The significance of differences between groups was verified by Duncan's test. In the present study the addition of conjugated linoleic acid or sunflower oil did not impact the composition and amounts of saturated or unsaturated fatty acids in Longissimus dorsi muscle. © Leibniz Institute for Farm Animal Biology, Dummerstorf, Germany. Source


Wasilewski P.D.,University of Technology and Life Sciences in Bydgoszcz | Nowachowicz J.,University of Technology and Life Sciences in Bydgoszcz | Michalska G.,University of Technology and Life Sciences in Bydgoszcz | Bucek T.,University of Technology and Life Sciences in Bydgoszcz | And 2 more authors.
Annals of Animal Science | Year: 2012

The aim of the study was to investigate the impact of feeding pigs with different levels of conjugated linoleic acid (CLA) or sunflower oil (SFO) on the backfat fatty acid profile. The subjects of research were 60 crossbred gilts divided into 6 groups, which were fed different levels of conjugated linoleic acid or sunflower oil (0.5, 1.0 and 2.0%, respectively). All fatteners were kept and fed under standardized conditions. Animals were slaughtered at 95 kg of body weight. Fatty acid profile was determined in samples of backfat from each animal using gas chromatography. The significance of differences between groups was verified by Duncan's test. In the present study, a beneficial effect of adding CLA was that it reduced saturated fatty acids and increased unsaturated fatty acids in the adipose tissue (backfat) of pigs as compared to fatteners receiving SFO. The amount of monounsaturated fatty acids in the backfat decreased with increasing amounts of CLA or SFO. Modifying the fatty acid profile of pig backfat through addition of CLA or SFO would be beneficial to the health of consumers because relatively large amounts of fat pork, including backfat, are added to the sausages. Source

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