Transfer and toxicokinetic modeling of non-dioxin-like polychlorinated biphenyls (ndl-PCBs) into accidentally exposed dairy cattle and their calves - A case report.

Polychlorinated biphenyls (PCBs) are persistent environmental pollutants that accumulate in tissues of exposed animals and humans. This case report refers ton=3 dairy cows accidentally exposed to non-dioxin-like PCBs (ndl-PCBs) of unknown origin on a German farm. At study start they had a cumulative total of 122-643 ng/g fat in milk and 105-591 ng/g fat in blood, consisting mainly of PCBs 138, 153, and 180. Two cows calved during the study and their calves were raised on their mothers' milk, resulting in cumulative exposure until slaughter. A physiologically based toxicokinetic model was developed to describe the fate of ndl-PCBs in the animals. The toxicokinetic behavior of ndl-PCBs was simulated in individual animals, including transfer of contaminants into calves via milk and placenta. Both the simulations and experimental data indicate that contamination via both routes is significant. In addition, the model was used to estimate kinetic parameters for risk assessment.

Cause and Effect Analysis between Influencing Factors Related to Environmental Conditions, Hunting and Handling Practices and the Initial Microbial Load of Game Carcasses.

Environmental, hunting and handling factors affect the microbial load of hunted game and the resulting meat products. The aim of this study was to systematically investigate the influence of several factors on the initial microbial load (IML) of game carcasses during the early hunting chain. Eviscerated roe deer body cavities (n = 24) were investigated in terms of total viable count and the levels of Pseudomonas spp., Lactobacillus spp., Enterobacteriaceae and Escherichia coli (E. coli). Furthermore, a risk analysis based on the obtained original IML data, literature search and a Failure Mode and Effects Analysis (FMEA) was performed. The IML could be explained in a regression model by factors including the higher body weight (BW), damaged gastrointestinal tract by the shot, ambient temperature or rain. The levels of Lactobacillus spp. (p = 0.0472), Enterobacteriaceae (p = 0.0070) and E. coli (p = 0.0015) were lower on the belly flap surface when gloves were used during evisceration. The literature search revealed that studies examining influencing factors (IF) on the IML of game carcasses found contradictory effects of the comparable IF on IML. Potential handling failures may lead to a higher IML of game carcasses during the early hunting chain ranked by FMEA. Several handling practices for game carcasses are recommended, such as ensuring efficient cooling of heavier BW carcasses to limit bacterial growth or eviscerating heavier carcasses before lighter ones.

Marination increases the bioavailability of lead in game meat shot with lead ammunition.

As a consequence of the toxicological lead characteristics, a reduction of its exposure should consider all sources. Game meat might contain elevated levels of lead due to the use of lead ammunition. The aim of the present study was to investigate the effects of acidic marination on the bioavailability of ammunition-derived lead in game meat (Roe deer), using the growing pig as an animal model. Furthermore, the study should provide evidence that the large-area scattering of lead particles leads to noticeable differences in the individual lead intake per game meat portion. Pigs of group A (n 7) received lead-shot game meat, which was cooked in water. Pigs of group B (n 7) received lead-shot game meat, which was first marinated (wine and vinegar) and then cooked. The lead content of both game meat preparations was equal with 0⋅77-0⋅79 mg Pb/portion. Pigs of group C (n 4) received lead-free game meat, which was also marinated and cooked. Additionally, lead acetate was administered intravenously to group D pigs (n 4). Blood samples were taken on elevated time points before and after game meat intake/i.v.-application. The acidic marination increased the bioavailability of orally ingested lead, resulting in significantly higher blood lead concentrations. The bioavailability of lead was 2⋅7 % when game meat was just cooked and 15 % when the meat was marinated before. The considerable variation of the individual blood lead concentrations suggests that an inhomogeneous distribution of ammunition-derived lead particles (in terms of size and number) causes individually non-comparable lead intakes from the consumption of game meat.

Transfer of Per- and Polyfluoroalkyl Substances (PFAS) from Feed into the Eggs of Laying Hens. Part 2: Toxicokinetic Results Including the Role of Precursors.

A feeding study was performed to examine the bioaccumulation of per- and polyfluoroalkyl substances (PFAS) in laying hens' tissues and plasma and feed-to-egg transfer rates and half-lives. A 25 day exposure was followed by a 42 day depuration period. A target analysis revealed substantial amounts of the precursors N-methyl and N-ethyl perfluorooctane sulfonamidoacetic acid (Me- and EtFOSAA), perfluorooctane sulfonamidoacetic acid (FOSAA), and perfluorooctane sulfonamide (FOSA). In tissues and eggs, the highest bioaccumulation was found for PFHxS, PFHpS, PFOS, and PFOA. Low levels of PFHxS (all samples), PFOS, and FOSAA (in yolk) were measurable even after the depuration period. The egg elimination half-lives of PFOS and aforementioned precursors were estimated to be 4.3 days, while the transfer rates of PFOS and all precursors taken together were 0.99. The transfer rate of PFOA was around 0.49. PFHxS and PFHpS showed apparent transfer rates of >100%, which is hypothesized to indicate the presence of precursors.

Transfer of Per- and Polyfluoroalkyl Substances (PFAS) from Feed into the Eggs of Laying Hens. Part 1: Analytical Results Including a Modified Total Oxidizable Precursor Assay.

The group of per- and polyfluoroalkyl substances (PFAS) comprises thousands of chemicals, which are used in various industrial applications and consumer products. In this study, a feeding experiment with laying hens and feed grown on a contamination site was conducted, and PFAS were analyzed in the feed and eggs to assess the transfer of PFAS into eggs. A targeted analysis of perfluoroalkyl acids (PFAAs) and different sulfonamides was performed. Additionally, the total oxidizable precursor (TOP) assay was modified by fully oxidizing small amounts of the samples instead of oxidizing their extracts in order to overcome potential losses during extraction. Targeted analysis showed the presence of known PFAAs and four sulfonamides in the feed and egg yolk samples. In the plant-based feed, short-chain PFAAs, methyl and ethyl perfluorooctane sulfonamidoacetic acid (Me- and EtFOSAA), and perfluorooctane sulfonic acid (PFOS) were the most abundant PFAS. In the eggs, PFOS, FOSAA, and its alkylated homologues showed the highest concentrations. The TOP assay revealed the presence of substantial amounts of precursors with different chain lengths from C4 to C8. The highest relative increase of PFOA after oxidation was observed in egg yolk from the end of the exposure period (828%). The results of this study demonstrate the transfer of PFAAs and their precursors into hens' eggs and emphasize the contribution of (known and unidentified) precursors to the overall PFAS burden in edible products. The modified TOP assay approach was shown to be a powerful tool to better assess the total burden of samples with PFAS.

Physiologically based toxicokinetic models and in silico predicted partition coefficients to estimate tetrachlorodibenzo-p-dioxin transfer from feed into growing pigs.

Tetrachlorodibenzo-p-dioxin (TCDD) is a ubiquitous, toxic, persistent and bioaccumulative organic pollutant. TCDD can potentially enter the food chain through contaminated food of animal origin as a consequence of feed contamination. Prediction of the TCDD transfer from feed into animal products is thus important for human health risk assessment. Here, we develop several physiologically based toxicokinetic (PBTK) models of TCDD transfer from contaminated feed into growing pigs (Sus scrofa) exposed to doses ranging from 24.52 to 3269.25 ng of TCDD. We test the consequences of explicit dose-dependent absorption (DDA) versus the indirect effects of a self-induced liver metabolism (SIM). The DDA and SIM models showed similar fit to experimental data, although currently it is not possible to unequivocally make statement on a mechanistic preference. The performance of both toxicokinetic models was successfully evaluated using the 1999 Belgian case of contaminated fats for feeding. In combination with toxicokinetic models of other dioxin congeners, they can be used to formulate maximum allowance levels of dioxins in feedstuffs for pigs. Additionally, the implementation of in silico-predicted partition coefficients was explored as a useful alternative to predict TCDD tissue distribution in low-dose scenarios without recurring to animal experiments.

Lead content in wild game shot with lead or non-lead ammunition - Does "state of the art consumer health protection" require non-lead ammunition?

The toxicity of lead has been known for a long time, and no safe uptake level can be derived for humans. Consumers' intake via food should therefore be kept as low as possible. Game meat can contain elevated levels of lead due to the use of lead ammunition for hunting. A risk assessment conducted in 2010 by the German Federal Institute for Risk Assessment including various consumption scenarios revealed a possible health risk for extreme consumers of game meat hunted with lead ammunition (i.e. hunters and members of hunters' households). Babies, infants, children and women of childbearing age were identified as vulnerable group with regards to the developmental neurotoxicity of lead. It was noted, that a sound data base was required in order to refine the assessment. Therefore, the research project "Safety of game meat obtained through hunting" (LEMISI) has been conducted in Germany, with the aims of determining the concentrations of lead (as well as of copper and zinc) brought into the edible parts of game meat (roe deer (Capreolus capreolus) and wild boar (Sus scrofa)) due to using either lead or non-lead hunting ammunition, whilst concurrently taking geogenic (i.e. "background") levels of lead into account. Compared to non-lead ammunition, lead ammunition significantly increased lead concentrations in the game meat. The use of both lead and non-lead ammunition deposited copper and zinc in the edible parts of game meat, and the concentrations were in the range of those detected regularly in meat of farm animals. For the average consumer of game meat in Germany the additional uptake of lead only makes a minor contribution to the average alimentary lead exposure. However, for consumers from hunters' households the resulting uptake of lead-due to lead ammunition-can be several times higher than the average alimentary lead exposure. Non-lead bullets in combination with suitable game meat hygienic measures are therefore recommended in order to ensure "state of the art consumer health protection".

Copper and zinc content in wild game shot with lead or non-lead ammunition - implications for consumer health protection.

The aim of this study was to examine the contamination of game meat with copper and zinc and establish whether the use of alternative (non-lead) ammunition can lead to higher or unsafe levels of copper and zinc in the meat of roe deer, wild boar and red deer. The research project "Safety of game meat obtained through hunting" (LEMISI) was conducted in Germany with the purpose of examining the entry of lead as well as copper and zinc into the meat of hunted game when using either lead or non-lead ammunition. The outcome of this study shows that the usage of both lead-based ammunition and alternative non-lead ammunition results in the entry of copper and zinc into the edible parts of the game. Using non-lead ammunition does not entail dangerously elevated levels of copper and zinc, so replacing lead ammunition with alternative ammunition does not introduce a further health problem with regard to these metals. The levels of copper and zinc in game meat found in this study are in the range found in previous studies of game. The content of copper and zinc in game meat is also comparable to those regularly detected in meat and its products from livestock (pig, cattle, sheep) for which the mean human consumption rate is much higher. From the viewpoint of consumer health protection, the use of non-lead ammunition does not pose an additional hazard through copper and zinc contamination. A health risk due to the presence of copper and zinc in game meat at typical levels of consumer exposure is unlikely for both types of ammunition.

Perfluoroalkylated substances in edible livers of farm animals, including depuration behaviour in young sheep fed with contaminated grass.

Perfluoroalkylated substances (PFASs) present a potential health risk for consumers. In animals these compounds are known to accumulate in livers. In order to determine potential PFASs contamination in commercially available livers, samples from farmed sheep, horses, cows, pigs and chicken were collected from the Dutch market. PFOS was the only detectable PFAS and its concentration was higher in free ranging animals like cows and sheep. The detected levels of PFOS in the liver samples were very low (up to 4.5 ng g(-1) ww). To further study the kinetic behaviour in foraging animals, samples from a study in which sheep were fed with grass obtained from a river floodplain, were examined. PFOS was the only detectable PFAS in the contaminated grass pellets, showing a level of about 0.5 µg kg(-1). Young blackhead sheep were fed with either clean or contaminated grass for a period up to 112 days. A time-dependent increase in liver PFOS concentrations was observed from 2.4 to 10.9 ng g(-1) ww after 8 and 112 days respectively. A time-dependent depuration was observed in livers of animals switched to clean grass after 56 days of exposure, from 9.2 to 4.7 ng g(-1) ww after 64 and 112 days respectively. The percentage of PFOS ingested from the grass and retained in the liver was estimated to be 12% at day 56, and decreased gradually to 6% after 56 days on clean grass, showing that the decrease in levels is not only caused by an increase in liver weight. Levels detected in commercial livers but also those in the sheep study would not lead to exceedance of the current TDI for PFOS set by EFSA. Therefore, it can be assumed that they do not present a risk for human health.

Regulatory toxicology in the twenty-first century: challenges, perspectives and possible solutions.

The advent of new testing systems and "omics"-technologies has left regulatory toxicology facing one of the biggest challenges for decades. That is the question whether and how these methods can be used for regulatory purposes. The new methods undoubtedly enable regulators to address important open questions of toxicology such as species-specific toxicity, mixture toxicity, low-dose effects, endocrine effects or nanotoxicology, while promising faster and more efficient toxicity testing with the use of less animals. Consequently, the respective assays, methods and testing strategies are subject of several research programs worldwide. On the other hand, the practical application of such tests for regulatory purposes is a matter of ongoing debate. This document summarizes key aspects of this debate in the light of the European "regulatory status quo", while elucidating new perspectives for regulatory toxicity testing.

Concentrations of strontium, barium, cadmium, copper, zinc, manganese, chromium, antimony, selenium, and lead in the liver and kidneys of dogs according to age, gender, and the occurrence of chronic kidney disease.

This study was conducted to measure the concentrations of strontium (Sr), barium (Ba), cadmium (Cd), copper (Cu), zinc (Zn), manganese (Mn), chromium (Cr), antimony (Sb), selenium (Se), and lead (Pb) in canine liver, renal cortex, and renal medulla, and the association of these concentrations with age, gender, and occurrence of chronic kidney disease (CKD). Tissues from 50 dogs were analyzed using inductively coupled plasma mass spectrometry. Cu, Zn, and Mn levels were highest in the liver followed by the renal cortex and renal medulla. The highest Sr, Cd, and Se concentrations were measured in the renal cortex while lower levels were found in the renal medulla and liver. Female dogs had higher tissue concentrations of Sr (liver and renal medulla), Cd (liver), Zn (liver and renal cortex), Cr (liver, renal cortex, and renal medulla), and Pb (liver) than male animals. Except for Mn and Sb, age-dependent variations were observed for all element concentrations in the canine tissues. Hepatic Cd and Cr concentrations were higher in dogs with CKD. In conclusion, the present results provide new knowledge about the storage of specific elements in canine liver and kidneys, and can be considered important reference data for diagnostic methods and further investigations.

Accumulation of polychlorinated dibenzo-p-dioxins, dibenzofurans, and biphenyls in livers of young sheep.

A major part of sheep livers contains levels of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) and polychlorinated biphenyls (PCBs) above the former but to some extent also the new maximum levels (MLs) in the EU. In order to investigate the relationship between the intake of these contaminants and their accumulation in livers, kidney fat and meat, young blackhead sheep were fed with grass pellets containing PCDD/Fs at 2.5 times the maximum level. Levels of PCDD/Fs in livers were already quite high at the start of the exposure but increased 3-fold within 56d, exceeding the new product based MLs. Levels in meat and fat also increased but did not exceed the MLs. Although less elevated in the grass, both dl- and ndl-PCB levels also increased in liver and fat. Their kinetics in the tissues was less clear, potentially caused by increased levels in the straw given to the sheep during the whole experimental period. There was a clear difference in the behavior of the various congeners, the PCDFs and especially the higher chlorinated PCDFs and PCDDs showing a higher accumulation in the liver. In the case of the PCBs, this was particularly true for PCB 126. When switched to clean grass after 56d, the levels in livers and other tissues decreased to about the levels in the control sheep within 56d. This offers a potential solution for decreasing the intake of consumers.

Concentrations of strontium, barium, cadmium, copper, zinc, manganese, chromium, antimony, selenium and lead in the equine liver and kidneys.

The concentrations of specific elements in the equine liver and kidneys are of practical relevance since horses are not only food-producing animals, but also partially serve as an indicator for the environmental pollution, as the basic feed includes plants like grass, grain and fruits. In this study, the concentrations of strontium (Sr), barium (Ba), cadmium (Cd), copper (Cu), zinc (Zn), manganese (Mn), chromium (Cr), antimony (Sb), selenium (Se) and lead (Pb) were measured in the liver, renal cortex and renal medulla of 21 horses (8 male; 13 female; aged between 5 months-28 years), using inductively coupled plasma mass spectrometry. Comparable Cu and Zn concentrations were detected in the liver and renal cortex, while approximately 50% lower concentrations were measured in the renal medulla. The lowest Sr, Cd and Se, but the highest Mn, Sb and Pb concentrations were measured in the liver. The Ba concentrations were comparable in the renal cortex and medulla, but lower in the liver of the horses. Gender-related differences were observed for Cd, Mn and Cr, with higher Cd concentrations in the liver, but lower Mn concentrations in the renal cortex and lower Cr concentrations in the renal medulla of female horses. Age-related differences were detected for most measured elements, however, the animal number per age-group was only low. In conclusion, the present study provides important reference data for the storage of Sr, Ba, Cd, Cu, Zn, Mn, Cr, Sb, Se and Pb in the liver and kidneys of horses, which are of practical relevance for an evaluation of the exposure of horses to these elements, either via feed or the environment.

Liver and kidney concentrations of strontium, barium, cadmium, copper, zinc, manganese, chromium, antimony, selenium and lead in cats.

BACKGROUND: In order to provide new knowledge on the storage of strontium (Sr), barium (Ba), cadmium (Cd), copper (Cu), zinc (Zn), manganese (Mn), chromium (Cr), antimony (Sb), selenium (Se) and lead (Pb) in the feline organism, we measured the concentrations of these elements in the liver, renal cortex and renal medulla, evaluating also the impact of age, sex or the occurrence of a chronic kidney disease (CKD). The element concentrations in the tissues of 47 cats (22 male; 25 female; aged between 2 months and 18 years) were measured using inductively coupled plasma mass spectrometry. RESULTS: Cu, Zn and Mn were the highest in the liver, followed by the renal cortex and the renal medulla. The Cd concentrations were lower in the renal medulla compared to the renal cortex and the liver, and Sr was higher in the renal medulla compared to the liver. The Se concentrations in the cortex of the kidneys were higher than in the medulla of the kidneys and in the liver. Higher Cd concentrations were measured in the renal cortex of female cats, while no further gender-related differences were observed. Except for Cr, Sb and Se, age-dependencies were detected for the storage of all elements. The occurrence of a CKD also affected the storage of the elements, with lower concentrations of Ba (renal medulla), Zn (renal cortex; renal medulla) and Mn (liver; renal medulla), but higher Cd concentrations (liver; renal cortex) in diseased cats. CONCLUSIONS: In conclusion, the present results provide new information on the accumulation of specific elements in the feline liver and kidneys, demonstrating a dependency on age and an impaired kidney function, but not on the sex of the animals.

Toxicokinetics of seven perfluoroalkyl sulfonic and carboxylic acids in pigs fed a contaminated diet.

The transfer of a mixture of perfluoroalkyl acids (PFAAs) from contaminated feed into the edible tissues of 24 fattening pigs was investigated. Four perfluoroalkyl sulfonic (PFSAs) and three perfluoroalkyl carboxylic acids (PFCAs) were quantifiable in feed, plasma, edible tissues, and urine. As percentages of unexcreted PFAA, the substances accumulated in plasma (up to 51%), fat, and muscle tissues (collectively, meat 40-49%), liver (under 7%), and kidney (under 2%) for most substances. An exception was perfluorooctanesulfonic acid (PFOS), with lower affinity for plasma (23%) and higher for liver (35%). A toxicokinetic model is developed to quantify the absorption, distribution, and excretion of PFAAs and to calculate elimination half-lives. Perfluorohexanoic acid (PFHxA), a PFCA, had the shortest half-life at 4.1 days. PFSAs are eliminated more slowly (e.g., half-life of 634 days for PFOS). PFAAs in pigs exhibit longer elimination half-lives than in most organisms reported in the literature, but still shorter than in humans.

A probabilistic model for the carry-over of PCDD/Fs from feed to growing pigs.

When food producing animals are contaminated with PCDD/F congeners, information on the contaminant's concentration in the bodies of the animals at time of slaughter is needed for risk management purposes. We have developed a mathematical model for the kinetics of PCDD/Fs in growing pigs in case of contaminated feed fed for a limited duration of time. This model allows the prediction of concentrations in body fat. It considers absorption fractions of PCDD/Fs, clearance by metabolism, dilution by growth and excretion through fecal fat. The model parameters were calibrated by fitting the model to experimental data. On the basis of this toxicokinetic model a probabilistic model has been constructed. The probabilistic model handles the parameters with appropriate probability distributions and Monte-Carlo simulation technique, providing for realistic situations with many animals and a range of contaminations and feeding intervals. We applied the new model to describe the German dioxin incident of winter 2010/2011 and discuss its viability as decision tool. The approach demonstrated here is a showcase how a risk assessment in the case of contaminated feeding can be performed.

Absorption, distribution, and milk secretion of the perfluoroalkyl acids PFBS, PFHxS, PFOS, and PFOA by dairy cows fed naturally contaminated feed.

The transfer of the perfluoroalkyl acids (PFAAs) perfluorobutanesulfonate (PFBS), perfluorohexanesulfonate (PFHxS), perfluorooctanesulfonate (PFOS), and perfluorooctanoate (PFOA) from feed into tissue and milk of dairy cows was investigated. Holstein cows (n = 6) were fed a PFAA-contaminated feed for 28 days. After the PFAA-feeding period, three cows were slaughtered while the others were fed PFAA-free feed for another 21 days (depuration period). For PFAA analysis plasma, liver, kidney, and muscle tissue, urine, and milk were sampled and analyzed using high-performance liquid chromatography (HPLC) with tandem mass spectrometry (MS/MS). The average daily intake of dairy cows was 3.4 ± 0.7, 4.6 ± 1.0, 7.6 ± 3.7 and 2.0 ± 1.2 µg/kg body weight (bw) for PFBS, PFHxS, PFOS, and PFOA, respectively. Overall, PFBS, PFHxS, PFOS, and PFOA showed different kinetics in dairy cows. In plasma, concentrations of PFBS (mean = 1.2 ± 0.8 µg/L) and PFOA (mean = 8.5 ± 5.7 µg/L) were low, whereas PFHxS and PFOS continuously increased during the PFAA-feeding period up to maximal concentrations of 419 ± 172 and 1903 ± 525 µg/L, respectively. PFOS in plasma remained constantly high during the depuration period. PFOS levels were highest in liver, followed by kidney, without significant differences between feeding periods. The highest PFHxS levels were detected in liver and kidney of cows slaughtered on day 29 (61 ± 24 and 98 ± 31 µg/kg wet weight (ww)). The lowest PFAA levels were detected in muscle tissue. At the end of the feeding study, cumulative secretion in milk was determined for PFOS (14 ± 3.6%) and PFHxS (2.5 ± 0.2%). The other two chemicals were barely secreted into milk: PFBS (0.01 ± 0.02%) and PFOA (0.1 ± 0.06%). Overall, the kinetics of PFOA were similar to those of PFBS and substantially differed from those of PFHxS and PFOS. The very low concentration of PFBS in plasma and milk, the relatively high urinary excretion, and only traces of PFBS in liver (0.3 ± 0.3 µg/kg ww) and kidney (1.0 ± 0.3 µg/kg ww) support the conclusion that PFBS does not accumulate in the body of dairy cows.

Colostrum and milk selenium, antioxidative capacity and immune status of dairy cows fed sodium selenite or selenium yeast.

Dietary selenium (Se) can be supplemented from organic or inorganic sources and this may affect Se metabolism and functional outcome such as antioxidative status and immune functions in dairy cows. A feeding trial was performed with 16 Holstein-Friesian dairy cows fed with a total mixed ration (0.18 mg Se/kg dry matter (DM)) either without Se supplement (Control, n = 5), or with Se from sodium selenite (Group SeS, n = 5) or Se yeast (Group SeY, n = 6). In Groups SeS and SeY, the Se supplementation amounted to an additional intake of 4 mg Se and 6 mg Se/d during gestation and lactation, respectively. The effect of both Se sources was characterised by milk Se and antioxidant levels, and the phenotyping and functional assessment of phagocytic activity of milk immune cells. Se yeast has been found to increase (p ≤ 0.001) the milk Se and antioxidant levels markedly compared to the control group. The experimental treatment did not affect the immune parameters of the cows. Lymphocyte subpopulations and phagocytosis activity of neutrophilic granulocytes were affected neither by the Se intake nor by the two different dietary supplements. It can be concluded that sodium selenite and Se yeast differ considerably in their effects on antioxidant status in dairy cows. However, the basal dietary Se concentration of 0.18 mg/kg DM seemed to be high enough for the measured immune variables.

Carryover of maduramicin from feed containing cross-contamination levels into eggs of laying hens.

Maduramicin is a coccidiostat authorized as feed additive in the European Union for chickens and turkeys for fattening but not for laying hens, considering the risk of residues in eggs. The unavoidable cross-contamination of non-target feed with coccidiostats is regulated by Commission Directive 2009/8/EC and resulting carry-over in food by Commission Regulation (EC) No. 124/2009. To verify the compliance of the maximum levels for maduramicin in feed (50 µg/kg) and eggs (2 µg/kg), the carry-over from feed into eggs was investigated. Diets containing 10, 30, and 50 µg of maduramicin/kg of feed were fed to laying hens. Feed, egg white, and yolk were analyzed by LC-MS/MS. Maduramicin residues were only detected in in egg yolk. Feeding the 10 µg/kg maduramicin diet resulted in maduramicin concentrations up to 2.5 µg/kg in whole eggs, already exceeding the maximum level. A carry-over rate of 8% maduramicin from feed into eggs was calculated.