Deposition and depletion of decoquinate in eggs after administration of cross-contaminated feed.

Decoquinate, a chemical coccidiostat used as a feed additive, can occur in eggs due to cross-contamination of feedstuffs for laying hens. An experiment was designed to assess the transfer of decoquinate to hen eggs and its distribution between egg yolk and egg white. Hens were given the feed containing decoquinate at a cross-contamination level (0.34 mg kg(-1)) and collected eggs were analysed using an LC-MS/MS method. The plateau level was reached on the eighth day of the experiment and averaged 8.91 µg kg(-1), which is far below the maximum level established at 20 µg kg(-1) for whole eggs. Decoquinate was deposited mostly in egg yolks (26.2 µg kg(-1)) and did not deplete completely during 14 days of administration of decoquinate-free feed. The results confirmed that administration of cross-contaminated feed is associated with very low risk of non-compliant residue levels of decoquinate in eggs.

Residue depletion of decoquinate in chicken tissues after oral administration.

A rapid, sensitive, and reliable high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed and validated for the analysis of decoquinate in chicken tissues. The compounds were extracted using acetonitrile by liquid-liquid extraction (LLE) and purified with an Oasis(™) HLB solid-phase extraction (SPE) cartridge. Chromatographic separation was performed on an XTerra C18 reversed-phase column with a mobile phase of water containing 0.1% formic acid and acetonitrile. The analyte was detected by tandem quadrupole mass spectrometry after positive electrospray ionization by multiple reaction monitoring. The detection and quantitation limits were 1 and 2.5 µg/kg, respectively. The recoveries of edible tissues ranged from 85.3% to 104.9%, with relative standard deviations (RSD) lower than 10.4%. The depletion profile of decoquinate was studied in healthy chickens after oral administration of feed containing 27.2 mg/kg decoquinate for 10 consecutive days. The residue concentrations of decoquinate in chicken muscle and liver were detected using the developed method. The highest residue concentrations were attained 0.25 day post-treatment, and decoquinate residues were still detected 5 days postmedication in the tissues examined. The developed method has been successfully applied to the depletion study of decoquinate in chicken tissues. The recommended withdrawal period with oral administration based on our research is 3 days.

The mode of action of anticoccidial quinolones (6-decyloxy-4-hydroxyquinoline-3-carboxylates) in chickens.

The anticoccidial mode of action of quinolones (6-decyloxy-4-hydroxyquinoline-3-carboxylates) against Eimeria tenella and E. acervulina in chickens has been investigated, using decoquinate and M&B 15,584 as examples. The well known static effect on sporozoites of relatively high continuous drug concentrations in the food masked other components of the mode of action, newly described here. Lower concentrations of quinolones allowed sporozoites to continue their development. First-stage schizonts were susceptible to a secondary cidal effect, although later schizonts seemed to be rather refractory. Furthermore, the sporulation of oocysts produced by E. tenella that completed its life cycle in the presence of suboptimal concentrations of quinolones was inhibited: this probably reflects a drug effect on gametocytes. Quinolones were absorbed rapidly from the chicken intestine, probably in less than 1 h. Drug withdrawal experiments showed that quinolones persisted in chicken tissues at active concentrations for up to 48 h. Despite their static effect on sporozoites, they may nevertheless be expected to exert a therapeutic effect against drug-sensitive coccidia in interrupted regimes that allow the later cidal effect to come into play. This allows immunity to coccidiosis to develop in the presence of drug. These new results, with the previously available data have been combined in an updated account of the anticoccidial mode of action of quinolones in the chicken.

In vivo expression of in vitro anticoccidial activity.

Large-scale screening has led to the identification of several experimental compounds that have very potent intrinsic activity against coccidia, but the lack of translation to in vivo efficacy has been a major hurdle in developing such leads into effective new drugs. We developed methods to explore the impact of oral availability and appropriate distribution in tissue, both of which are potentially important factors in the expression of activity in vivo. For the compounds that we examined, neither oral absorption nor distribution to the site of infection appeared to be the critical barrier to in vivo expression of intrinsic anticoccidial activity. Elucidation of the nature of additional factors that might be involved could assist greatly in the identification of useful new anticoccidial agents.

Metabolism of decoquinate in chickens and Japanese quail.

Thirty mature chicken hens and 40 mature Japanese quail hens were used in an experiment to compare pathways of decoquinate (DQ) excretion. Labelled DQ was injected into chickens (.5 microCi via wing vein puncture) and quail (.25 microCi via cardiac puncture) on Day 0. Blood was sampled at 0, 1.5, 3, 6, 9, 12, 24, and 48 h postinjection. Eggs and excreta of chickens and quail were collected for 28 and 14 days, respectively, and analyzed for 14C. Six chickens and eight quail were sacrificed prior to 14C-DQ injection and also on Days 1, 7, 14, and 34 or 32 postinjection. Samples of liver, heart, kidney, bile, skin, fat, and muscle were analyzed for 14C. Blood rapidly cleared 14C in both species, and the half-time of 14C excretion via excreta was more rapid in quail (.37 day) than in chickens (.92 day). Little 14C was found in the eggs of quail (.32% of dose) and chickens (.17% of dose). Quail appeared to excrete peak amounts of detectable 14C 1 day earlier (Day 4) than chickens (Day 5). Liver contained the greatest concentration of 14C on Day 1 in both species. By the end of the experiment, less than 1% of the dose remained in liver or other organs. Results indicate that chickens and quail metabolize 14C-DQ at comparable rates and by similar pathways.

Comparative drug depletion in domestic animals and birds.

Decoquinate (Rhone-Poulenc Inc) and Narasin (Eli Lilly and Co) were selected as model drugs for a comparison of metabolism between major (cattle and chickens) and minor (sheep and quail) species. Decoquinate has been studied in all four species. Narasin studies are in progress in chickens and quail. More than 96% of injected 14C-decoquinate (DQ) was eliminated from blood of all species within 1 hr. Disappearance of the remaining 1 to 4% from blood was rapid for all species. Half-times for DQ appearance in excreta were all less than one day. Cumulative excretion of DQ in eggs of chickens and quail was about 1% for both species. Disappearance of DQ from tissues was essentially complete in 14 days. More than 80% of injected 14C-narasin was eliminated from blood within 1/2 hr. Disappearance of the remainder was rapid for both chickens and quail.