Residues of sulfadiazine and doxycycline in egg matrices due to cross-contamination in the feed of laying hens and the possible correlation with physicochemical, pharmacokinetic and physiological parameters

In the poultry industry, the widespread use of veterinary drugs such as antimicrobial compounds may lead to the presence of residues in whole eggs, egg white and egg yolk. During this study, laying hens received experimental feed containing sulfadiazine or doxycycline at cross-contamination levels of 2.5%, 5% and 10% of the therapeutic concentration. Since the therapeutic dose is 250 mg kg(-1) for both substances, cross-contamination concentrations in the feed of 6.25, 12.5 and 25 mg kg(-1) were expected. Whole egg, egg white and egg yolk samples were collected during the treatment and depletion period and were analysed via liquid chromatography-tandem mass spectrometry. For both drugs, a plateau phase was reached within 3-5 days and residue concentrations were detected in all egg matrices. For the 10% cross-contamination group, residual sulfadiazine concentrations of 208, 299 and 60 μg kg(-1) and residual doxycycline concentrations of 455, 332, 206 μg kg(-1) were detected in whole egg, egg white and egg yolk on day 13 of the treatment period, respectively. Both sulfadiazine and doxycycline had higher concentrations in egg white than in egg yolk, but the egg white-egg yolk ratio was higher for sulfadiazine than for doxycycline. As neither drug is allowed in Belgium for use in laying hens, residues may pose food safety concerns.     

Residues of sulfadiazine and doxycycline in egg matrices due to cross-contamination in the feed of laying hens and the possible correlation with physicochemical,pharmacokinetic and physiological parameters

In the poultry industry, the widespread use of veterinary drugs such as antimicrobial compounds may lead to the presence of residues in whole eggs, egg white and egg yolk. During this study, laying hens received experimental feed containing sulfadiazine or doxycycline at cross-contamination levels of 2.5%, 5% and 10% of the therapeutic concentration. Since the therapeutic dose is 250?mg?kg^?1 for both substances, cross-contamination concentrations in the feed of 6.25, 12.5 and 25?mg?kg^?1 were expected. Whole egg, egg white and egg yolk samples were collected during the treatment and depletion period and were analysed via liquid chromatography-tandem mass spectrometry. For both drugs, a plateau phase was reached within 3–5 days and residue concentrations were detected in all egg matrices. For the 10% cross-contamination group, residual sulfadiazine concentrations of 208, 299 and 60?µg?kg^?1 and residual doxycycline concentrations of 455, 332, 206?µg?kg^?1 were detected in whole egg, egg white and egg yolk on day 13 of the treatment period, respectively. Both sulfadiazine and doxycycline had higher concentrations in egg white than in egg yolk, but the egg white–egg yolk ratio was higher for sulfadiazine than for doxycycline. As neither drug is allowed in Belgium for use in laying hens, residues may pose food safety concerns.     

Doxycycline and sulfadimethoxine transfer from cross-contaminated feed to chicken tissues

During feed preparation at feed mills or during feed mixing in bins at farms, the accidental contamination of feed at trace levels by veterinary drug residues, commonly known as carry-over, can accidentally but frequently occur. To evaluate the concentrations of residual antimicrobials in poultry edible tissues, due to contaminated feed, sulfadimethoxine and doxycycline were administered for 10 days to chickens in poultry feed incurred at the contamination levels frequently found during national feed monitoring programmes (1–5?mg?kg^?1). Sulfadimethoxine and doxycycline residual concentrations detected in muscle (<LOD and 31?µg?kg^?1, respectively), liver (13 and 56?µg?kg^?1, respectively) and kidney (56 and 115?µg?kg^?1, respectively) were compared with their maximum residue limits (MRLs) fixed by EC 470/2009 and EU 37/2010 Regulations for a preliminary risk evaluation.     

Evaluation of the EEC four-plate test and Premi test for screening antibiotic residues in trout (Salmo trutta)

The four‐plate test (FPT) method was used for the determination of sulfadiazine/trimethoprim residues in trout in the European Community (EC). This microbiological inhibition test used three media seeded with Bacillus subtilis at different pH values (6, 7.2 or 8.0) and a fourth medium seeded with Micrococcus luteus. The Premi test was also used for the determination of sulfadiazine/trimethoprim residues in trout. The effects of a trout muscle supernatant on detection limits of antibiotics with microbiological inhibition tests were measured. The supernatant was applied directly on top of the paper disks impregnated with aqueous antibiotic solutions. Inhibition zones were compared with those obtained by the same standard solution of sulfadiazine/trimethoprim without the trout supernatant. The detection limit of penicillin, sulfadimidine, streptomycin groups of antibiotics were determined for checking the test plates. For supernatant from fish fed with pellets containing antibiotics (sulfadiazine/trimethoprim), the inhibition zones increased according to the duration of the feed application. In the control group, no inhibition zones were detected. The concentration of the residues accumulated and reached a plateau after 5 days. The antibiotics were detectable in the same concentration as on day 7, but three days later (on day 10) they were no longer detectable in the fish samples. In contrast to the ‘FPT’, 3 days after discontinuing the medicated diet, there were still residues detectable by ‘Premi test’ on day 10. Data is presented that shows that the FPT and the Premi test methods are very useful for the determination of sulfadiazine/trimethoprim residues in trout.     

Probabilistic risk model to assess the potential for resistance selection following the use of anti-microbial medicated feed in pigs

The cross-contamination of non-medicated feed with residues of anti-microbials (AM) causes a public and animal health concern associated with the potential for selection and dissemination of resistance. To analyse the associated risks, a probabilistic model was built using @Risk® (Palisade Corporation®) to show the potential extent of the effect of cross-contaminated pig feed on resistance selection. The results of the model include estimations of the proportion of pigs per production stage with residues of doxycycline, chlortetracycline, sulfadiazine and trimethoprim in their intestinal contents, as a result of exposure to cross-contaminated feed with different carry-over levels, in Belgium. By using a semi-quantitative approach, these estimations were combined with experimental data on AM concentrations associated with potential for resistance-selection pressure. Based on this model, it is estimated that 7.76% (min = 1.67; max = 36.94) of sows, 4.23% (min = 1.01%; max = 18.78%) of piglets and 2.8% (min = 0.51%; max = 14.9%) of fatteners in Belgium have residues of doxycycline in their intestinal tract due to consumption of feed with at least 1% carry-over. These values were estimated to be almost triple for sulfadiazine, but substantially lower for chlortetracycline and trimethoprim. Doxycycline concentrations as low as 1 mg/L (corresponding to consumed feed with at least 1% carry-over) can select for resistant porcine commensal Escherichia coli in vitro and in vivo. Conclusions on this risk could not be drawn for other AM at this stage, due to the lack of data on concentrations associated with resistance development. However, since the possibility of resistance mechanisms (e.g. co-selection) occurring cannot be excluded, the results of this model highlight that the use of AM medicated feed should be minimised where possible. In case of medicated feed production, good practice should be followed thoroughly at all levels of production, distribution, storage and administration, with a special focus on the feed distributed to piglets and sows.     

Depletion of tylosin residues in feathers,muscle and liver from broiler chickens after completion of antimicrobial therapy

Tylosin is one of the most commonly used antimicrobial drugs from the macrolide family and in broiler chickens it is used specially for the treatment of infectious pathologies. The poultry industry produces several by-products, among which feathers account for up to 7% of a chicken’s live weight, thus they amount to a substantial mass across the whole industry. Feathers have been repurposed as an animal feed ingredient by making them feather meal. Therefore, the presence of high concentrations of residues from antimicrobial drugs in feathers might pose a risk to global public health, due to re-entry of these residues into the food chain. This work aimed to characterise the depletion behaviour of tylosin in feather samples, while considering its depletion in muscle and liver tissue samples as a reference point. To achieve this goal, we have implemented and validated an analytical methodology suitable for detecting and quantifying tylosin in these matrices. Sixty broiler chickens, raised under controlled conditions, received an oral dose of 32 mg kg^-1 of tylosin for 5 days. Tylosin was quantified in muscle, liver and feathers by liquid chromatography coupled with a photodiode array detector (HPLC-DAD). High concentrations of tylosin were detected in feather samples over the whole experimental period after completing both the therapy and the recommended withdrawal time (WDT). On the other hand, tylosin concentrations in muscle and liver tissue samples fell below the limit of detection of this method on the first sampling day. Our results indicate that the WDT for feather samples is 27 days, hence using feather meal for the formulation of animal diets or for other agricultural purposes could contaminate with antimicrobial residues either other livestock species or the environment. In consequence, we recommend monitoring this matrix when birds have been treated with tylosin, within the context of poultry farming.     

Screening for antibiotic residues in the trout by the Four Plate test,Premi test and ELISA test

The effectiveness of various assay systems for the examination of trout was investigated. The Four Plate test (FPT), Premi test and ELISA test were used for determining sulfadiazine/trimethoprim residues in trout. According to the results of the Four Plate test, when trouts were fed with pellets containing antibiotics (sulfadiazine/trimethoprim), the inhibition zones increased according to duration of feed application. In the control group, no inhibition zones were detected. The concentration of the residues accumulated and reached a plateau after 5 days. The antibiotics were detectable in the same concentration as on day 10, but 5 days later (on day 15) they were no longer detectable in the trout by EEC FPT. In contrast to the FPT, 5 days after discontinuing the medicated diet, there were still residues detectable by the Premi test and ELISA test on day 15. According to the ELISA test results of the trout; from day 1 to day 10 the results of sulfadiazine residues were found to be 4.18, 8.02, 9.13, 20.10, 21.32, 20.52, 20.75, 20.99, 20.90, 20.32 ppb, respectively. On day 10, the antibiotics were detectable in the same concentration. On day 15, sulfadiazine residues were found to be 6.82 ppb, but 5 days later (on day 20) they were no longer detectable in the trout. EEC Four Plate method presented less sensitivity and effectiveness in the detection of sulfadiazine.     

Tissue residues of clopidol (3,5-dichloro-2,6-dimethyl-4-pyridinol) in chickens in relation to withdrawal times

Broiler chickens were fed on a commercial diet containing 0.0125% clopidol for 34 days. They were killed 0-10 days after withdrawal of the premedicated feed and clopidol was determined in liver and muscle samples by a sensitive gas-liquid chromatography (g.l.c.) method. During the first two post-withdrawal days the clopidol concentration in the liver decreased rapidly from 7 to 0.5 mg/kg and the level in muscle declined from 3 to 0.1 mg/kg. There was little decline in the clopidol concentrations from days 2 to 10, the levels during this period being 0.2-0.8 mg/kg in liver and 0.05-0.2 mg/kg in muscle. In addition to the above experimental study, liver and muscle samples collected at a Swedish slaughterhouse from broiler chickens raised on clopidol-containing feed were analysed for residues of this drug. Large variations were found in the clopidol levels in broilers from different producers. The levels in the liver ranged from 0.05 to 8.0 mg/kg and those in muscle from 0.03 to 3.5 mg/kg. The present results emphasize the need to carry out field studies to check the levels of feed additive residues in edible tissues from chickens.     

武夷岩茶抗疲劳抗氧化作用研究

在20只小鼠和20只大鼠上分别做武夷岩茶抗疲劳和抗氧化作用试验。对照组饲喂基础饲料和自来水;试验组添加3%的武夷岩茶,饮0.5%的茶汤;试验期3周。抗疲劳试验结果:对照组死亡时间为1.90±0.66(min);试验组死亡时间为2.52±0.77(min),比对照组显著延长(p<0.05)。抗氧化试验结果:试验组大鼠血清、心肌、骨骼肌、肾脏的SOD比对照组升高13%、37%、39%、20%;GSH-Px比对照组升高7%、64%、32%、48%(均p<0.05);而血清、骨骼肌、肝脏、肾脏的MDA分别比对照组降低47%、32%、29%、35%(p<0.05或0.01);证明武夷岩茶有显著抗疲劳和抗氧化作用。     

氟苯尼考在鸡蛋和蛋鸡组织中的残留规律及预测模型建立

氟苯尼考是一种被广泛应用于动物养殖过程中的抗生素药物，可能残留于畜产品中，被人类长期食用会对机体造成耐药性、免疫抑制等不良影响。本实验旨在探讨蛋鸡摄入不同剂量氟苯尼考后，鸡蛋及各组织中氟苯尼考及其代谢产物氟苯尼考胺的清除规律，建立残留预测数学模型。本实验选取处于产蛋高峰期的罗曼粉壳蛋鸡250 只（350 日龄、体质量（1.97±0.07）kg），随机分为5 组，每组50 只，分别给予氟苯尼考0（对照）、30、60、120、240 mg/（kg mb·d），连续给药5 d。采用液相色谱-串联质谱法测定不同休药时间点蛋黄、蛋清、卵黄、肌肉、肝脏中氟苯尼考及其代谢产物氟苯尼考胺的残留含量。结果表明：休药1～3 d时，240 mg/（kg mb·d）剂量组蛋鸡产蛋率与其他处理组相比显著降低（P＜0.05），休药4 d后恢复至对照组水平；氟苯尼考及其代谢产物氟苯尼考胺在产蛋鸡组织中残留含量分布：蛋黄＞卵黄＞蛋清＞肝脏＞肌肉；不同组织残留氟苯尼考及氟苯尼消除所需休药时间：肝脏＜肌肉＜蛋清＜蛋黄＜卵黄；氟苯尼考给药剂量和休药时间对残留含量影响高度显著（P＜0.001），且给药剂量与休药时间互作效应高度显著（P＜0.001）。休药时间、给药剂量和残留含量之间呈现二元二次回归关系（P＜0.001）。为达到《中国兽药典（2015年版）》所规定的氟苯尼考给药剂量范围（40～60 mg/（kg mb·d）），肌肉等组织需休药1 d、蛋清蛋黄需休药13 d、卵黄需休药21 d。     

Determination of glycarbylamide residues in chicken tissues by HPLC with post-column reaction

A newly developed HPLC method for determination of glycarbylamide (GB) in chicken liver was applied to other tissues (muscle, fat and kidney). The recoveries of GB in muscle, fat and kidney were 87.2% (CV 0.5), 91.3% (CV 4.7), 79.7% (CV 1.0), respectively. The detection limit of GB was 0.01 ppm. GB concentrations were determined by this method in tissues (muscle, fat, liver and kidney) from chickens sacrificed 5 days after oral administration of GB mixed in feed at 60 mg/kg of feed for 7 days. GB was not detected in these chicken tissues.     

Determination of lasalocid residues in the tissues of broiler chickens by liquid chromatography-tandem mass spectrometry

Lasalocid is a polyether ionophoric coccidiostat used for the prevention of coccidiosis in poultry at a prescribed concentration and during a certain time interval. Due to a public health concern about the presence of coccidiostat residues in poultry, the aim of the present study was to determine the levels of lasalocid residues in the edible tissues of broiler chickens (breast muscle, thigh muscle, heart, liver, gizzard, kidneys and skin/fat) fed commercially produced feed containing 100?mg?kg^?1 of lasalocid in complete feed throughout the 5-day withdrawal period (WP). The residues were investigated by liquid chromatography coupled with electrospray ionisation (ESI) tandem mass spectrometry (MS/MS) with triple quadrupole. The limit of detection (LOD) and the limit of quantification (LOQ) of the method were 0.47 and 1.44?µg?kg^?1, respectively. The average recovery based on the matrix-fortified calibrations for chicken tissues ranged between 79% and 98%. Lasalocid was found to accumulate in the liver, followed by the heart, skin/fat, kidneys, thigh muscle and gizzard. The lowest concentrations of lasalocid residues were found in the breast muscle. On day 5 of the WP, residue concentrations of lasalocid did not decline below the LOQ of the method, but were far below the maximum residue level (MRL) established for lasalocid in poultry from 20 to 100?µg kg^?1 by European Commission Regulation (EU) No. 37/2010. The results confirmed that the WP established for lasalocid is sufficient to ensure the decline of its residues in the tissues of broiler chickens to the safe residue level.     

Depletion study of enrofloxacin and its metabolite ciprofloxacin in edible tissues and feathers of white leghorn hens by liquid chromatography coupled with tandem mass spectrometry

To ensure delivery of safe foods to consumers, withdrawal times for drugs must be respected according to the maximum residual limits established by regulatory agencies. Because of availability and price, feather meal is currently incorporated into animal feed as a protein source for farm species. Few data are available on residual drugs in feathers from treated animals. A depletion study was performed with laying hens treated intramuscularly with 5% enrofloxacin (Enromic) at 10 mg/kg body weight over 3 days. Thirty-three birds were treated and slaughtered at different times between 6 and 216 h after treatment; and samples of muscle plus skin, liver, kidney, and feathers were collected. High-performance liquid chromatography coupled with a tandem mass spectrometry method was validated before sample analysis to determine the decision limit, detection capability, recovery, and precision. Liver was the edible tissue with the slowest drug depletion. A withdrawal time of 6 days was calculated based on European Union maximum residual limits (100 microg/kg). A withdrawal time of 9 days was calculated based on Japan maximum residual limits (10 microg/kg). Enrofloxacin plus ciprofloxacin concentrations in feathers remained high through all sampling periods. Thus, feathers from treated animals should not be fed to food-producing animals.     

Determination of lasalocid residues in the tissues of broiler chickens by liquid chromatography-tandem mass spectrometry

Lasalocid is a polyether ionophoric coccidiostat used for the prevention of coccidiosis in poultry at a prescribed concentration and during a certain time interval. Due to a public health concern about the presence of coccidiostat residues in poultry, the aim of the present study was to determine the levels of lasalocid residues in the edible tissues of broiler chickens (breast muscle, thigh muscle, heart, liver, gizzard, kidneys and skin/fat) fed commercially produced feed containing 100?mg?kg?1) of lasalocid in complete feed throughout the 5-day withdrawal period (WP). The residues were investigated by liquid chromatography coupled with electrospray ionisation (ESI) tandem mass spectrometry (MS/MS) with triple quadrupole. The limit of detection (LOD) and the limit of quantification (LOQ) of the method were 0.47 and 1.44?μg?kg?1, respectively. The average recovery based on the matrix-fortified calibrations for chicken tissues ranged between 79% and 98%. Lasalocid was found to accumulate in the liver, followed by the heart, skin/fat, kidneys, thigh muscle and gizzard. The lowest concentrations of lasalocid residues were found in the breast muscle. On day 5 of the WP, residue concentrations of lasalocid did not decline below the LOQ of the method, but were far below the maximum residue level (MRL) established for lasalocid in poultry from 20 to 100?μg kg?1 by European Commission Regulation (EU) No. 37/2010. The results confirmed that the WP established for lasalocid is sufficient to ensure the decline of its residues in the tissues of broiler chickens to the safe residue level.     

Transfer of the coccidiostats monensin and lasalocid from feed at cross-contamination levels to whole egg,egg white and egg yolk

Recent legislation has addressed the unavoidable carry-over of coccidiostats and histomonostats in feed, which may lead to the presence of residues of these compounds in eggs. In this study, laying hens received cross-contaminated feed at a ratio of 2.5%, 5% and 10% of the therapeutic dose of monensin and lasalocid for broilers. The eggs were collected during the treatment and depletion period and were analysed using liquid chromatography-tandem mass spectrometry. The different egg matrices were separated and analysed during the plateau phase. High lasalocid concentrations, which exceeded the maximum residue level, and low monensin concentrations were found in whole egg. Plateau levels were reached at days 7–9 for lasalocid and at days 3–5 for monensin. For lasalocid, the highest concentrations were measured in egg yolk; residue concentrations in egg white were very low.     

Liquid chromatography tandem mass spectrometry determination of maduramycin residues in the tissues of broiler chickens

Maduramycin is a polyether ionophoric coccidiostat used to prevent coccidiosis in poultry at a prescribed concentration over a certain time interval. Due to public health concerns about the presence of coccidiostat residues in poultry, the aim of the present study was to determine the level of maduramycin residues in the tissues of broiler chickens fed commercially produced feed containing 5 mg kg^?1 of maduramycin in complete feed throughout the 5-day withdrawal period (WP). The residues were investigated by liquid chromatography (LC) coupled with electrospray ionisation (ESI) tandem mass spectrometry (MS/MS). The limit of detection (LOD) and limit of quantification (LOQ) of the method were 0.3 and 0.8 µg kg^?1, respectively. The average recovery based on matrix-fortified calibrations for chicken tissues was 90%. Maduramycin was found to be rapidly distributed in all tissues. The highest concentrations of maduramycin residues were found in the heart followed by the skin, liver, gizzard, kidneys and, finally, muscle (thigh and breast). On day 5 of the WP, residue concentrations of maduramycin did not decline below the LOQ of the method. Our results emphasize the need to establish a maximum residue limit (MRL) for maduramycin to control its residue levels in edible tissues from chickens before slaughter.     

Detection of maduramycin residues in the tissues of chickens and pheasants by the screening test for antibiotic residues (STAR)

Maduramycin is a coccidiostat authorized as a feed additive in poultry. Council Directive 96/23/EC stipulates that monitoring of foods of animal origin for residues of coccidiostats is mandatory. The aim of the present study was to evaluate the STAR for the screening of maduramycin residues in the tissues of broiler chickens and pheasants. Both animal species were supplied feed medicated with Cygro l% premix according to recommendations for use (5 mg kg(-1) of complete feed). The residues were investigated for a period of 7 days: day 0 (the last day of the administration of maduramycin), days 1-5 (the days of the withdrawal period) and day 6 (the first day after elapse of the withdrawal period). According to STAR the positivity of the sample (the presence of residues of antibacterial substances) is indicated by a zone of inhibition exceeding 2 or 4 mm in width, depending on the test organism. Maduramycin residues were detected only on the plates seeded with the test organism Bacillus stearothermophilus var. calidolactis ATCC 10149. The results showed that there was higher potential for the presence of maduramycin residues in broiler chickens than in pheasants. All chicken tissues (muscle/thigh and breast/gizzard, liver, heart, kidneys, spleen, lungs) were positive for maduramycin (inhibition zones ≥4 mm) not only throughout the withdrawal period, but also even 5 days after elapse of the withdrawal period. In the case of pheasants the positive results were detected in the gizzard, liver, heart, kidneys, spleen and lungs. On day 5 of the withdrawal period no positive results were detected; however, on day 6 the heart and spleen were positive again.     

Biomagnification study on organochlorine compounds in marine aquaculture: the sea bass (Dicentrarchus labrax) as a model

Biomagnification of organochlorine compounds (pesticides and polychlorinated biphenyls) through the marine aquaculture food chain is investigated. From first-feeding, specimens of sea bass were exposed to commercial fish feed (that contained DDTs and PCBs residues) for ca. 24 months, and selected tissues (white and red muscle, liver, and visceral fat) were analyzed after 6 and 24 months of diet exposure. Data obtained showed that experimental fish tissues presented a similar contamination pattern to that of fish feed, and biomagnification processes of these compounds were proved. Additionally, commercial sea bass cultured in farms from the western Mediterranean were analyzed, their organochlorine concentrations being significantly lower than those of the 24 month old experimental fish. Thus, the exposition of human population to OCs through consumption of cultured fish would be lower than expected from experimental biomagnification studies, although red muscle presented similar OC levels in both cases, which were much higher than those of white muscle. Although levels of organochlorine compounds were found to be low, the persistence, ubiquity, and toxicity of these compounds, together with their presence in fish feed, make it necessary to monitor OC residues in the routine quality assurance programs of aquaculture activities, as this food chain is a source of these toxic compounds for human consumers. The development of sensitive analytical methodology based on GC-MS/MS has allowed for the reaching of low detection limits required to carry out the present study.     

Effect of L-carnitine infusion and feed restriction on carnitine status in lactating Holstein cows

Previously we determined that abomasal infusion of l-carnitine increased in vitro hepatic fatty acid oxidation, decreased liver lipid accumulation, and supported higher fat-corrected milk yield in feed-restricted lactating cows. The objectives of this study were to examine the effects of supplemental l-carni-tine and amount of feed intake on free carnitine and carnitine ester concentrations in liver, muscle, milk, and plasma of lactating dairy cows. Eight lactating Holstein cows (132 ± 36 d in milk) were used in a replicated 4 × 4 Latin square design with 14-d periods to test factorial combinations of water or l-carnitine infusion (20 g/d; d 5 to 14) and ad libitum or restricted (50% of previous 5-d intake; d 10 to 14) dry matter intake. Plasma was obtained 3 times daily on d 4, 8, and 12; milk samples were collected on d 8, 9, 13, and 14. Liver and muscle were biopsied on d 14 of each period. Free carnitine, short-chain acylcarnitine, and long-chain acylcarnitine concentrations were determined using a radioenzymatic assay coupled with ion exchange chromatography. Abomasal l-carnitine infusion increased total carnitine in plasma on d 8 and d 12. All liver carnitine fractions were increased by carnitine infusion. Feed restriction elevated concentrations of free carnitine, long-chain acylcarnitine, and total carnitine in liver tissue from carnitine-infused cows but not in those infused with water. In muscle, acid-soluble carnitine, long-chain acylcarnitine, and total carnitine concentrations were increased by carnitine infusion and feed restriction without significant interaction. Feed restriction increased free carnitine concentrations in muscle from water-infused cows but not in carnitine-infused cows. Carnitine infusion increased the concentration of each milk carnitine fraction as well as milk carnitine output on d 8 to 9. On d 13 to 14, all carnitine fractions except short-chain acylcarnitine were increased in milk from water-infused, feed-restricted cows, whereas all fractions were increased in carnitine-infused, feed-restricted cows. Carnitine infusion increased total carnitine in plasma, liver, muscle, and milk during feed restriction, whereas feed restriction alone increased carnitine concentrations in muscle and milk but not in liver. Liver carnitine concentrations might limit hepatic fatty acid oxidation capacity in dairy cows during the periparturient period; therefore, supplemental l-carnitine might decrease liver lipid accumulation in periparturient cows.     

Dose-response feeding study of short chain chlorinated paraffins (SCCPs) in laying hens: effects on laying performance and tissue distribution, accumulation and elimination kinetics

Technical short chain chlorinated paraffins (C10-C13 with 60% chlorine) were fed to 93 laying hens from 24 to 32 weeks of age in increasing concentrations of up to 100 mg/kg feed. No significant influence on health, relative organ weights or performance (laying intensity, egg weight, feed consumption) was noted. The chlorinated paraffin content of the tissues was linearly related to the concentration of short chain paraffins of the feed. The highest concentrations were found in abdominal fat, egg yolk and fatty tissues. Breast muscle, egg albumen and bile fluid contained minimal or no residues. Less than 1% of the chlorinated paraffins ingested were incorporated into the body (without head, feet, gut and feathers), whereas about 1.5% were eliminated with the egg yolk and 30% were excreted with urine and faeces. A six-week kinetic depuration study revealed a biphasic elimination with half-lifes of 4-40 min (liver, kidneys, legs, fat, blood) for the initial rapid phase, and 15-30 days (blood, fat, liver, yolk, kidneys, legs) for the terminal slow phase.