Pasteurella sp. associated with fatal septicaemia in six African elephants.

The sudden mortality of African elephants (Loxodonta africana) in Botswana and Zimbabwe in 2020 provoked considerable public interest and speculation. Poaching and malicious poisoning were excluded early on in the investigation. Other potential causes included environmental intoxication, infectious diseases, and increased habitat stress due to ongoing drought.

Assessing the influence of pig slurry pH on the degradation of selected antibiotic compounds.

Veterinary medicines are routinely used in animal husbandry and the environment may consequently be exposed to them via manure applications. This presents potential environmental and societal risks such as toxicological effects to aquatic/terrestrial organisms and the spread of antimicrobial resistance. Regulatory studies that assess the degradability of veterinary antibiotics during manure storage currently permit the use of just one manure per animal type although we speculate that heterogenic properties such as pH could be driving significant variability within degradation rates.

The determination of monomers and oligomers from polyester-based can coatings into foodstuffs over extended storage periods.

The polymeric coating used in metal packaging such as cans for foods and beverages may contain residual amounts of monomers used in the production of the coating, as well as unreacted linear and cyclic oligomers. Traditionally, although designed for use with plastic food contact materials, food simulants have been used to determine the migration of monomers from coatings into foodstuffs. More recently, food simulants have also been used to determine oligomeric species migrating from can coatings.

Occurrence of polybrominated diphenylethers, hexabromocyclododecanes, bromophenols and tetrabromobisphenols A and S in Irish foods.

The occurrence of polybrominated diphenyl ethers (PBDEs), hexabromocyclododecanes (HBCDs), tetrabromobisphenol A (TBBPA) and other phenolic brominated flame retardants (BFRs) in Irish foodstuffs has been assessed. A total of 53 food samples including eggs, milk, fish, fat and offal were tested. Eighty-one percent of the samples contained at least one measurable PBDE congener.

Metabolite profiling using liquid chromatography/quadrupole time-of-flight mass spectrometry for the identification of a suitable marker and target matrix of griseofulvin use in bovines.

Rationale: Griseofulvin is an antifungal agent with potential for misuse in food-producing animals. Little is known about its metabolism in ruminants and hence what are suitable marker residues and target matrices for monitoring purposes.

Methods: Tissues harvested from cattle treated with the antifungal agent griseofulvin were screened using liquid chromatography coupled to positive and negative electrospray ionization (ESI) quadrupole time-of-flight mass spectrometry (qToFMS) operated in ToF mode.

Toggled RNA aptamers against aminoglycosides allowing facile detection of antibiotics using gold nanoparticle assays.

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.

Analysis of incurred crystal violet in Atlantic salmon (Salmo salar L.): comparison between the analysis of crystal violet as an individual parent and leucocrystal violet and as total crystal violet after oxidation with 2,3-dichloro-5,6-dicyanobenzoquinone.

Due to on-going concern about the occurrence of triphenylmethane dye residues in fish destined for human consumption, a depletion study of crystal violet in salmon was carried out. Atlantic salmon less than 12 months old were exposed to crystal violet in fresh water at 15°C and subsequently sampled at 1, 7, 14, 28, 63 and 91 days after exposure. The salmon were then analysed by two analytical methods.

Screening method for the analysis of antiviral drugs in poultry tissues using zwitterionic hydrophilic interaction liquid chromatography/tandem mass spectrometry.

A screening method for the analysis of seven anti-viral drugs in poultry tissue has been developed. These include anti-influenza drugs (amantadine, rimantadine, zanamivir and oseltamivir and its carboxylate metabolite), anti-herpes drugs (acyclovir and ganciclovir) and an immunomodulator (imiquimod). Poultry tissue was extracted in acetonitrile:water:acetic acid.

An RNA-aptamer-based assay for the detection and analysis of malachite green and leucomalachite green residues in fish tissue.

A robust screening assay employing solid phase extraction (SPE) followed by a novel aptamer-based procedure is presented for the rapid detection and semiquantitation of the triphenylmethane dye, Malachite Green (MG) and its primary metabolite Leucomalachite Green (LMG) in fish tissue. To the authors' knowledge, this is the first reported use of an RNA aptamer for the development of a diagnostic assay for the detection of chemical residues in food. The aptamer based screening assay is found to be highly specific for MG; but has negligible affinity for the LMG metabolite.

Multi-residue determination of phenolic and salicylanilide anthelmintics and related compounds in bovine kidney by liquid chromatography-tandem mass spectrometry.

This paper describes an analytical method for four phenolic and salicylanilide anthelmintics authorised for use within the EU (nitroxinil, oxyclozanide, rafoxanide and closantel) in bovine kidney, and the extension of this procedure to include a number of related compounds; ioxynil, niclosamide, salicylanide and 3-trifluoromethyl-4-nitrophenol (TFM). The method comprises a solvent extraction with 1% acetic acid in acetone and clean-up using a mixed-mode anion-exchange solid phase extraction column. Determination is by reversed phase LC-MS/MS.

Detection of glucocorticoid bioactivity in bovine urine samples using a reporter gene assay.

The illegal use of anabolic substances in the meat producing industry is an ongoing problem due to the continual production of new synthetic compounds and/or the practice of low-level cocktail administration to avoid detection by the surveillance schemes of EU member states National Plan surveillance systems. We present a highly sensitive reporter gene assay and sample extraction procedure based on a two step solid phase extraction and high performance liquid chromatography, developed for the detection of glucocorticoid abuse in bovine urine. The assay is capable of detecting compounds with glucocorticoid bioactivity and is extremely sensitive with an EC(50) of 0.

Multiresidue determination of triarylmethane and phenothiazine dyes in fish tissues by LC-MS/MS.

The occurrence of residues of malachite green and its leuco-metabolite in tissues of farmed fish for human consumption have long been of concern and there is extensive literature on methods of analysis and surveillance for these compounds. Recently, concern has been expressed that the use of other related compounds in place of malachite green may go undetected. This paper describes a new method for extending the range of triarylmethane and related phenothiazine dyes that can be detected in fish.

Analysis of androgenic steroid Girard P hydrazones using multistage tandem mass spectrometry.

Seven androgenic steroids have been converted into steroid hydrazones using Girard P hydrazine and analysed by electrospray ionisation multistage tandem mass spectrometry. The cationic derivatives 17alpha-testosterone hydrazone, 17beta-nortestosterone hydrazone, 17beta-bolasterone hydrazone, 17alpha-boldenone hydrazone, 17beta-fluoxymesterone hydrazone, 17alpha-trenbolone hydrazone and 4-chloroandrosten-3,17-dione hydrazone show good response in positive ion mode with enhancements for the method of up to 33 times relative to the native species. Detailed characterisation of fragmentation pathways reveals structurally specific ions formed by fragmentation of the hydrazine moiety.

Verification of compliance with organic meat production standards by detection of permitted and nonpermitted uses of veterinary medicines (tetracycline antibiotics).

In the production of "organic" meat, one of the controlled processes is the use of veterinary drugs. Strict standards are in place as to when and how such drugs may be used. Therefore, the aim of this project was to determine whether it was possible to distinguish between a single therapeutic dose of a tetracycline (permitted under the standards) and both multiple therapeutic dosing and prophylactic dosing (not permitted).

The effect of cooking on veterinary drug residues in food: nicarbazin (dinitrocarbanilide component).

The change of concentration of residues of the marker compound for the anti-coccidial drug nicarbazin, N,N'-bis(4-nitrophenyl)urea (dinitrocarbanilide, DNC), was investigated in model oil and aqueous solutions and in chicken muscle and egg. In model aqueous solutions, DNC decreased rapidly in concentration upon heating followed by a much more gradual decomposition. The curves produced when this information was plotted were not typical of exponential decay.

Meeting maximum residue limits: an improved screening technique for the rapid detection of antimicrobial residues in animal food products.

A rapid, high-throughput antimicrobial screening assay was developed using either a physical fluid extraction or a solvent extraction technique coupled to the commercially available PremiTest. The solvent extraction approach was fully validated for a wide range of tissues and the fluid extraction approach partially validated for porcine muscle. Both procedures can detect a wide range of antimicrobial compounds at or below maximum residue limit concentrations.

Improved clean-up for the determination of lasalocid in 'difficult' food matrices.

Methodology has been developed for the determination of lasalocid in analytically 'difficult' matrices such as processed and spiced foods. The procedure was based on an existing silica-based solid-phase extraction (SPE) clean-up to which was added a novel NH2 SPE step before HPLC with fluorescence detection. Use of the additional step enabled the determination of lasalocid in matrices such as baby food, meat pies ('pasties'), etc.

Screening and confirmation of triphenylmethane dyes and their leuco metabolites in trout muscle using HPLC-vis and ESP-LC-MS.

An extraction and clean-up protocol for the determination of Malachite Green and Crystal Violet and the corresponding leuco compounds in trout muscle has been developed. Final determination is by HPLC with visible (screening) or ESP-MS (confirmation) detection. In both cases lead(IV) oxide was used on-line to oxidise the leuco compounds back to the parent after chromatographic separation and prior to detection.

Screening of sulphonamides in egg using gas chromatography-mass-selective detection and liquid chromatography-mass spectrometry.

A rapid extraction and clean-up procedure for sulphonamide antibiotics in eggs suitable for both GC-MSD and LC-MS end determinations has been developed. The drugs were extracted using acetonitrile, acidified using acetic acid and cleaned-up using cation- and anion-exchange. For determination by GC-MSD, extracts were derivatised with diazomethane followed by pentafluoropropionic acid anhydride.

Effects of extraction and spiking procedures on the determination of incurred residues of oxytetracycline in cattle kidney.

The effects of different extraction and spiking procedures on the determination of incurred oxytetracycline residues in animal tissues have been investigated. The extraction procedures investigated--direct aqueous or organic solvent extraction, enzymic digestion or sonication--all gave similar results for incurred oxytetracycline concentration in cattle kidney after correction for spike recovery. There was therefore no evidence for binding or conjugation of oxytetracycline in this tissue.

Aspects of extraction, spiking and distribution in the determination of incurred residues of chloramphenicol in animal tissues.

Studies of distribution, extraction procedures and spiking protocols in the determination of incurred chloramphenicol residues in animal tissues have been carried out. An extraction procedure involving glucuronidase enzyme digestion was found to extract 10 times more incurred chloramphenicol from pig kidney than direct extraction without digestion. However, neither protease digestion nor ultrasonic probe treatment resulted in improved chloramphenicol extraction.

Improved method for the on-line metal chelate affinity chromatography-high-performance liquid chromatographic determination of tetracycline antibiotics in animal products.

An improved on-line metal chelate affinity chromatography-high-performance liquid chromatography (MCAC-HPLC) method for the determination of tetracycline antibiotics in animal tissues and egg has been developed. Extraction was carried out with ethyl acetate. The extract was then evaporated to dryness and reconstituted in methanol prior to on-line MCAC clean-up and HPLC-UV determination.

Determination of penicillins in animal tissues at trace residue concentrations: II. Determination of amoxicillin and ampicillin in liver and muscle using cation exchange and porous graphitic carbon solid phase extraction and high-performance liquid chromatography.

A method for the determination of the basic penicillins, amoxicillin and ampicillin, in animal tissue is described. The method used aqueous extraction with tungstate to remove proteins followed by cation exchange solid phase extraction (SPE) clean-up. This extract was further purified using porous graphitic carbon (PGC) SPE.

On-line metal chelate affinity chromatography clean-up for the high-performance liquid chromatographic determination of tetracycline antibiotics in animal tissues.

An on-line high-performance liquid chromatographic (HPLC) method for the determination of tetracycline, oxytetracycline, chlortetracycline and demeclocycline using metal chelate affinity chromatography-reversed-phase HPLC has been developed. The drugs were extracted with succinate buffer and the extract diluted with EDTA-pentanesulphonate buffer. Diluted extract was then absorbed onto a C8 or XAD-2 solid-phase extraction (SPE) cartridge and eluted with methanol.