Review: Bioavailability of trace elements in farm animals: definition and practical considerations for improved assessment of efficacy and safety.

Currently, the authorisation procedure of trace elements as feed additives in the European Union according to Regulation (EC) No. 1831/2003 does not consider the bioavailability of trace element sources. This manuscript provides framework conditions for in vivo experiments that aim to estimate differences in the relative bioavailability between supplements of essential trace elements. Framework conditions encompass necessary technical information on the test substance, the experimental design and diet composition as well as the suitability of status parameters that allow for relative comparisons of regression variables. This manuscript evolves recommendations for researchers to conduct solid and reliable experiments on the matter as well as decision makers to interpret the value of studies submitted with authorisation applications regarding a certain trace element supplement.

Transfer of Non-Dioxin-Like Polychlorinated Biphenyls (ndl-PCBs) from Feed and Soil into Hen Eggs.

Understanding the transfer of non-dioxin-like polychlorinated biphenyls (ndl-PCBs) into foods of animal origin is crucial for human health risk assessment. In two experiments, we investigated the transfer of ndl-PCBs from contaminated feed and soil into eggs and meat of laying hens. The transfer from the feed was investigated with 30 laying hens. The treated hens were divided into two groups fed a contaminated diet (12.8 µg/kg sum of indicator ndl-PCBs; 88% dry matter (DM)) for 28 and 63 days, respectively, and then experienced a depuration period of 100 days with control feed. The transfer from soil was investigated with 72 laying hens kept in three separate outdoor pens (with three levels of ndl-PCB soil contamination) for 168 days. In both experiments, eggs were collected and analyzed for ndl-PCBs. In the second experiment, animals (n = 3 at the beginning, n = 6 per group after 42, 84, and 168 days) were slaughtered to determine ndl-PCBs in meat (breast muscle tissue) fat. The transfer of ndl-PCB from both feed and soil was clearly measurable and concentrations in eggs quickly exceeded maximum levels. Clear differences between individual congeners were observed. In particular, the low-chlorinated ndl-PCBs 52 and 101 are hardly found in eggs, despite their relatively high concentration in feed and soil. PCBs 138, 153, and 180, on the other hand, were found in large proportions in eggs and meat.

Bioavailability of phthalate and DINCH® plasticizers, after oral administration of dust to piglets.

For decades, phthalates have been widely used as plasticizers in a large number of consumer products, leading to a complex exposure to humans via ingestion, inhalation or dermal uptake. Children may have a higher unintended dust intake per day compared to adults. Therefore, dust intake of children could pose a relevant exposure and subsequently a potential health risk. The aim of this study was to determine the relative bioavailability of certain phthalates, such as di(2-ethylhexyl) phthalate (DEHP), di-isononyl phthalate (DINP) and the non-phthalate plasticizer diisononyl 1,2-cyclohexanedicarboxylic acid (DINCH®, Hexamoll®), after ingestion of dust. Seven 5-week-old male piglets were fed five different dust samples collected from daycare centers. Overall, 0.43 g to 0.83 g of dust sieved to 63 µm were administered orally. The piglets' urine was collected over a period of 38 h. The excreted metabolites were quantified using an LC-MS/MS method. The mean uptake rates of the applied doses for DEHP, DINP, and DINCH® were 43% ± 11%, 47% ± 26%, and 9% ± 3.5%, respectively. The metabolites of DEHP and DINP showed maximum concentrations in urine after three to five hours, whereas the metabolites of DINCH®, reached maximum concentrations 24 h post-dose. The oral bioavailability of the investigated plasticizers was higher compared to the bioaccessibility reported from in vitro digestion tests. Furthermore, the bioavailability of DEHP did not vary substantially between the dust samples, whereas a dose-dependent saturation process for DINP was observed. In addition to other intake pathways, dust could be a source of plasticizers in children using the recent intake rates for dust ingestion.

Excretion pathways and ruminal disappearance of glyphosate and its degradation product aminomethylphosphonic acid in dairy cows.

From 6 balance experiments with total collection of feces and urine, samples were obtained to investigate the excretion pathways of glyphosate (GLY) in lactating dairy cows. Each experiment lasted for 26d. The first 21d served for adaptation to the diet, and during the remaining 5d collection of total feces and urine was conducted. Dry matter intake and milk yield were recorded daily and milk and feed samples were taken during the sampling periods. In 2 of the 6 experiments, at the sampling period for feces and urine, duodenal contents were collected for 5d. Cows were equipped with cannulas at the dorsal sac of the rumen and the proximal duodenum. Duodenal contents were collected every 2h over 5 consecutive days. The daily duodenal dry matter flow was measured by using chromium oxide as a volume marker. All samples (feed, feces, urine, milk and duodenal contents were analyzed for GLY and aminomethylphosphonic acid (AMPA). Overall, across the 6 experiments (n=32) the range of GLY intake was 0.08 to 6.67mg/d. The main proportion (61±11%; ±SD) of consumed GLY was excreted with feces; whereas excretion by urine was 8±3% of GLY intake. Elimination via milk was negligible. The GLY concentrations above the limit of quantification were not detected in any of the milk samples. A potential ruminal degradation of GLY to AMPA was derived from daily duodenal GLY flow. The apparent ruminal disappearance of GLY intake was 36 and 6%. In conclusion, the results of the present study indicate that the gastrointestinal absorption of GLY is of minor importance and fecal excretion represents the major excretion pathway. A degradation of GLY to AMPA by rumen microbes or a possible retention in the body has to be taken into account.

Inter- and intra-individual differences in serum copper and zinc concentrations after feeding different amounts of copper and zinc over two lactations.

The influence of different copper and zinc contents in rations on blood serum concentrations was tested in a feeding trial at the Institute of Animal Nutrition of the Federal Research Institute for Animal Health (FLI). A total of 60 German Holstein breed cows (38 first lactating and 22 second lactating cows) were tested. All animals received a diet based on maize- and grass silage ad libitum. The animals were divided into two groups with 30 cows each. Group A received concentrates supplemented with copper and zinc as recommended, whereas Group B was offered a concentrate with roughly double the amount of copper and zinc. Blood samples were taken several times during the lactation. The concentrations of copper and zinc in the serum were measured and correlations between trace element intake and their serum concentrations were calculated. The mean dry matter intake was significantly higher for cows in the second lactation (19.1 kg/day) than for cows in the first lactation (16.4 kg/day), whereas the DM-intake was constant in the different feeding groups (17.3 and 17.4 kg/day). The correlation between feed intake in the first and the second lactation was r = 0.76. The copper concentration in the serum was not influenced by the different feedings (11.9 and 12.5 micromol/l), but high variations were found between the cows. The influence of the lactation number was not significant (12.1 and 12.4 micromol/l). In contrast to copper, the zinc concentration in the serum was significantly higher for Group B (14.0 micromol/l) than for Group A (12.2 micromol/l), respectively, for second lactating cows (13.8 micromol/l) in relation to 12.7 micromol/l for first lactating cows. The individual variation was also high as for copper. Neither the correlation between copper intake and copper serum concentration nor the correlation between zinc intake and zinc serum concentration was significant.

Effect of several doses of zeolite A on feed intake, energy metabolism and on mineral metabolism in dairy cows around calving.

The object of the present study was to determine the influence of different zeolite A doses on dry matter intake (DMI) and mineral metabolism, and to evaluate an optimum dosage for preventing hypocalcaemia. Eighty pregnant dry cows were assigned to four groups (I-IV). They were fed a total mixed ration (TMR) ad libitum. Groups II, III and IV received an average daily dose of 12, 23 and 43 g zeolite A/kg DM for the last 2 weeks prepartum. Individually DMI was recorded daily. Blood and urine samples were taken before, during and after zeolite A supplementation. Serum was analysed for Ca, Mg, P(i), K, non-esterified fatty acids and beta-hydroxybutyrate (BHB). Urine was analysed for Ca, Mg, P(i), K and net acid-base excretion (NABE). After calving, milk yield (fat corrected milk) and milk composition were determined. During zeolite A supplementation, mean DMI of Group IV (7.3 +/- 1.3 kg/cow/day) was significantly lower compared to Groups I-III (10.1, 10.9, 9.5 kg/cow/day). The reduced feed intake of Group IV resulted in significantly increased BHB as well as decreased NABE after calving. Zeolite A supplementation in higher doses (III and IV) had a stabilizing effect on Ca metabolism around calving for older cows, whereas cows in Groups I and II showed a subclinical hypocalcaemia. The mean serum Mg concentration decreased significantly in older cows in Group IV at calving. The mean P(i) concentration in cows of Group IV decreased into ranges of hypophosphataemia already 1 week after beginning of zeolite A feeding. The mean DMI postpartum as well as the milk yield was not affected by zeolite A supplementation. Feeding of 23 g zeolite A/kg DM TMR prepartum proved to be an adequate dosage for reducing subclinical hypocalcaemia frequency without significant effects on feed intake and P(i) concentration in serum.

Effects of the Fusarium toxin deoxynivalenol on tissue protein synthesis in pigs.

The Fusarium toxin deoxynivalenol is known to be a potent protein synthesis inhibitor according to research in different in vitro and in vivo systems. However, the significance of this toxic feature was not yet evaluated for the pig. Therefore, an experiment was carried out to measure porcine tissue protein synthesis employing the so-called flooding dose technique using [(2)H(5)]-phenylalanine as tracer. A total of 25 male castrated pigs (initial weight of 23kg) were used. Seventeen pigs were fed the control diet (control) and eight the Fusarium toxin-contaminated diet (chronic oral deoxynivalenol (DON)) for approximately 4wks. Pigs recruited from the control group were treated with acute oral (n=7) and intravenous (i.v.) DON (n=5), and were exposed to the toxin once on the day of protein synthesis measurement. The DON-dose given prior to the beginning of protein synthesis measurement amounted to 2, 77, 83 and 53microg DONkg(-1) live weight for the control group, chronic oral DON-group, acute oral DON-group and acute i.v. DON-group, respectively. Protein synthesis expressed as fractional synthesis rate (FSR) was significantly reduced in kidneys, spleen and ileum of DON-exposed pigs with effects being less pronounced in the acute oral DON-group. FSR of liver, skeletal and heart muscle, mesenteric lymph nodes, duodenum, jejunum, jejunal mucosa cells, pancreas and lung were not affected by DON.