Clay minerals, such as layered double hydroxide (LDH) and montmorillonite (MMT), have attracted a great deal of attention for biological applications. Along with the rapid development of nanotechnology, public concern about the potential toxicity of nanoparticles is growing. In the present work, cytotoxicity of LDH and MMT was assessed in terms of inhibition of cell proliferation, generation of oxidative stress, and induction of inflammation response. Moreover, the biokinetics of LDH and MMT were evaluated; biokinetics provides information about in vivo absorption, distribution, and excretion kinetics. The results demonstrated that both LDH and MMT inhibited cell proliferation at relatively large concentrations and after long exposure time compared to other inorganic nanoparticles, although they generated reactive oxygen species (ROS). LDH induced pro-inflammatory cytokines in a size-dependent manner. Biokinetic study revealed that, after single-dose oral administration to mice, both LDH and MMT had extremely slow oral rates of absorption and did not accumulate in any specific organ. All the results suggest great potential of clay minerals for biological application at safe levels.

The objective of this study was to evaluate the effects of oak tannin extract (OTE) added in forage before ensiling on dairy cows fed at 92% of their digestible protein requirements. Six multiparous lactating Holstein cows were used in a crossover design (two treatments × two periods). The control treatment (CON) was based on a diet including 50% of grass silage, whereas the experimental treatment (TAN) included grass silage sprayed with OTE (26 g/kg DM) just before baling. Milk yield (on average 24 kg fat protein corrected milk per day) was not affected, but both milk and rumen fatty acids profiles were impacted by OTE. Nitrogen intake (415 g N per cow per day) and nitrogen use efficiency (NUE; 0.25 on average) were not affected, but a shift from urine (−8% of N intake relatively to control, P = 0.06) to faecal N (+5%; P = 0.004) was observed with the TAN diet (P ≤ 0.05). Nitrogen apparent digestibility was thus reduced for TAN (−3%; P ≤ 0.05). The effect of OTE on ruminal and milk FA profiles suggests an impact on rumen microbiota. Nitrogen isotopic discrimination between animal proteins and diet (Δ15N) was evaluated as a proxy for NUE. While no differences in NUE were observed across diets, a lower Δ15N of plasma proteins was found when comparing TAN v. CON diets. This finding supports the concept that Δ15N would mainly sign the N partitioning at the metabolic level rather than the overall NUE, with the latter also being impacted by digestive processes. Our results agree with a N shift from urine to faeces, and this strategy can thus be adopted to decrease the environmental impact of ruminant protein feeding.

Mesotrione is an effective herbicide for volunteer potato control but is not selective in several crops, including onion and carrot. Studies were conducted in 2006 to evaluate the effectiveness of wiper-applied mesotrione for control of volunteer potato in a potato crop. Surprisingly, nontreated potatoes growing adjacent to mesotrione-treated plants exhibited bleaching symptoms resembling mesotrione injury. Additional field trials confirmed injury to nontreated plants growing within 26 cm of potatoes treated with mesotrione applied with a wiper at 0.25, 0.5, and 1% (v/v) solutions. Greenhouse bioassays confirmed that mesotrione applied to potato leaves moved down through the plant and was exuded into perlite potting medium in sufficient quantities to injure potato plants that were exposed to the leachate from the perlite pots. In tracer studies, 52% of 14C-labeled mesotrione applied to potato leaves was absorbed into the potato plant by 15 d and 15% of the absorbed 14C-mesotrione was exuded into the soil and soil leachate. Mesotrione applied to potato by the wick application method has potential to injure neighboring susceptible plants from root uptake of exuded mesotrione.

The effect of feeding pigs in a three-phase feeding (3PF) system or a daily-phase feeding (DPF) system on growth performance, body composition, and N and P excretions was studied on 8 pens of 10 pigs each. Feeds for the 3PF and DPF treatments were obtained by mixing two feeds, one with a high nutrient concentration and the other with a low nutrient concentration. The DPF pigs tended (P=0.08) to consume more feed (+3.7%) than the 3PF pigs, but only during the first feeding phase. The DPF pigs consumed 7.3% less protein (P<0.01) but a similar amount of total P. For the whole growing period, the DPF pigs tended (P=0.08) to gain more weight (+2.4%) than the 3PF pigs, mainly because of faster growth (P=0.02) during the first feeding period. At the end of the experiment, total body protein mass was similar in the two treatment groups, but the DPF pigs had 8% more body lipids (P=0.04) than the 3PF pigs. Daily multiphase feeding reduced N excretion by 12% (P<0.01) but did not significantly reduce P excretion. In addition, feed costs, nutrient intake and nutrient excretion under the two feeding strategies were simulated and compared after different approaches were used to formulate complete feeds for each phase of the 3PF system, as well as the two feeds used in the DPF program. Simulated feed intake and growth was similar to those observed in the animal experiment. In comparison with the simulated 3PF system, the feed cost for the DPF pigs was reduced by 1.0%, the simulated N and P intakes were reduced by 7.3% and 4.4%, respectively, and the expected N and P excretions were reduced by 12.6% and 6.6%, respectively. The concomitant adjustment of the dietary concentration of nutrients to match the evaluated requirements of pig populations can be an efficient approach to significantly reduce feeding costs and N and P excretions in pig production systems.

Bumblebees need to keep bodily homeostasis and for that have an efficient system of excretion formed by the Malpighian tubules, ileum, and rectum. We analyzed the excretory organs of Bombus morio, a bee without rectal pads. In addition, we analyzed the rectal epithelium of Melipona quadrifasciata anthidioides which has rectal pads. The Malpighian tubules exhibited two cell types and the ileum four types. However, comparative analysis of the rectum showed that only cells of the anterior region of the rectal epithelium of B. morio are structurally distinct. We suggest that cells of the Malpighian tubules of B. morio have an excretory feature and that cells of ileum have different functions, such as ion absorption and water, organic compound, and protein secretion. In addition, only the anterior region of the rectum of B. morio showed characteristic absorption. We suggest that Malpighian tubules participate in the excretion of solutes and that the ileum and rectal epithelium are responsible for homeostasis of water and solutes, compensating for the absence of rectal papillae. These results contribute to our understanding of the morphophysiology of the excretory organs of bees without rectal pads.

Human milk oligosaccharides (HMO) are complex glycans that are highly abundant in human milk, but not in infant formula. Accumulating data, mostly from in vitro and animal studies, indicate that HMO benefit the breast-fed infant in multiple ways and in different target organs. In vitro incubation studies suggest that HMO can resist the low pH in the infant's stomach and enzymatic degradation in the small intestine and reach the colon in the same composition as in the mother's milk. The oligosaccharide composition in faeces of breast-fed infants is, however, very different from that in the mother's milk, raising questions on when, where and how HMO are metabolised between ingestion and excretion. To answer some of these questions, we established a pulse-chase model in neonatal rats and analysed HMO profiles to track their composition over time in five consecutive equal-length intestinal segments as well as in serum and urine. The relative abundance of individual HMO changed significantly within the first 2 h after feeding and already in the segments of the small intestine prior to reaching the colon. Only 3′-sialyllactose, the major oligosaccharide in rat milk, and hardly any other HMO appeared in the serum and the urine of HMO-fed rats, indicating a selective absorption of rat milk-specific oligosaccharides. The present results challenge the paradigm that HMO reach the colon and other target organs in the same composition as originally secreted with the mother's milk. The present results also raise questions on whether rats and other animals represent suitable models to study the effects of HMO.

A mathematical model was developed from literature data to predict the volume and composition of pig’s excreta (dry and organic matter, C, N, P, K, Cu and Zn contents), and the emission of greenhouse gases (CH4 and CO2) though respiration and from the intestinal tract, for each physiological stage (post-weaning and fattening pigs and lactating and gestating sows). The main sources of variation considered in the model are related to animal performances (feed efficiency, prolificacy, body weight gain, etc.), to water and nutrient intakes and to housing conditions (ambient temperature). Model predictions were validated by using 19 experimental studies, most of them performed in conditions close to those of commercial farms. Validation results showed that the model is precise and robust when predicting slurry volume (R2 = 0.96), slurry N (R2 = 0.91), P (R2 = 0.95) and to a lesser extent dry matter (R2 = 0.75) contents. Faeces and urine composition (minerals and macronutrients) can also be precisely assessed, provided the composition and the digestibility of the feed are well known. Sensitivity analysis showed strong differences in CH4 emission and excretion amounts and composition according to physiological status, animal performance, temperature and diet composition. The model is an efficient tool to calculate nutrient balances at the animal level in commercial conditions, and to simulate the effect of production alternatives, such as feeding strategy or animal performance, on excreta production and composition. This is illustrated by simulations of three feeding strategies, which demonstrates important opportunities to limit environmental risks through diet manipulations.

Oranges are rich sources of flavonoids that are bioactive and may protect against age-related diseases. The absorption of orange flavanones may be affected by factors such as processing and subject anthropometric variables, and the bioactivity of the absorbed phytochemicals depends on how they are metabolised during absorption. In a randomised cross-over study, twenty subjects consumed a single portion of orange fruit (150 g) or juice (300 g) that contained the flavanones narirutin and hesperidin, and an additional 109 subjects across a broad age range (18–80 years) consumed the juice. Flavanone metabolites were measured in regularly collected samples of plasma and urine. After consumption of fruit or juice, flavanone conjugates, but not the aglycones, were detected in plasma and urine. The flavanone conjugates were shown to include the 7- and 4′-O-monoglucuronides of naringenin, the 7- and 3′-O-monoglucuronides of hesperetin, two hesperetin diglucuronides and a hesperetin sulfo-glucuronide, but no aglycones or rutinosides. Analysis of the plasma pharmacokinetic and urinary excretion data on a dose-adjusted basis indicated no difference in absorption or excretion of either flavanone between the fruit and juice matrices. In the extended urinary excretion dataset the individual variation was very large (range 0–59 % urinary yield). There was a small but significant (P < 0·05) decrease in the excretion of hesperetin (but not naringenin) with increasing age (P < 0·05), but the effects of sex, BMI and contraceptive pill use were shown not to be associated with the variation in flavanone excretion.

It is essential to have a thorough knowledge of the bioavailability and metabolism of dietary flavonols to understand their role in disease prevention. Lightly fried onions containing 275μmol flavonols, principally quercetin-4′-glucoside and quercetin-3,4′-diglucoside, were fed to healthy human volunteers and plasma and urine were collected over a 24h period. Samples were analysed by HPLC with diode array and tandem mass spectrometric detection. Five flavonol metabolites, quercetin-3′-sulphate, quercetin-3-glucuronide, isorhamnetin-3-glucuronide, a quercetin diglucuronide and a quercetin glucuronide sulphate, were detected in plasma in quantifiable amounts with trace quantities of six additional quercetin metabolites. Sub-micromolar peak plasma concentrations (cmax) of quercetin-3′-sulphate, quercetin-3-glucuronide, isorhamnetin-3-glucuronide and quercetin diglucuronide were observed 0.6–0.8h after ingestion. In contrast, the cmax of quercetin glucuronide sulphate was 2.5h. The elimination half-lives (t1/2) of quercetin-3′-sulphate, quercetin-3-glucuronide and quercetin diglucuronide were 1.71, 2.33 and 1.76h respectively, while the t1/2 of isorhamnetin-3-glucuronide was 5.34h and that of quercetin glucuronide sulphate was 4.54h. The profile of metabolites excreted in urine was markedly different to that of plasma with many of the major urinary components, including quercetin-3′-glucuronide, two quercetin glucoside sulphates and a methylquercetin diglucuronide, absent or present in only trace amounts in the bloodstream indicative of substantial phase II metabolism. Total urinary excretion of quercetin metabolites was 12·9μmol, corresponding to 4·7% of intake. If these samples had been subjected to hydrolysis, as in many previous studies, only quercetin and isorhamnetin would have been detected and quantified. The bioactivity of these metabolites should be considered.

Rats were fed a grape seed extract (GSE) containing (+)-catechin, (−)-epicatechin and dimers, trimers, tetramers and polymeric procyanidins. Liver, kidney, brain and gastrointestinal (GI) tract together with plasma, urine and faeces were collected over a 24 h period and their flavan-3-ol content was analysed by HPLC with tandem mass spectrometry and diode array detection. Small amounts of the GSE flavan-3-ols moved out of the stomach and into the duodenum/jejunum, and to a greater extent the ileum 1 h after ingestion, and into the caecum after 2 h with relatively small amounts being detected in the colon after 3 h. The GI tract contained the parent GSE flavan-3-ols and procyanidins with only trace amounts of metabolites and there were no indications that proanthocyanidins were depolymerised in the GI tract releasing monomeric flavan-3-ols. Plasma contained exclusively catechin glucuronides and methylated glucuronide metabolites which were also detected in the liver and kidneys. These metabolites were also present in urine together with sulphated metabolites and low amounts of the procyanidin dimers B1, B2, B3 and B4 as well as the trimer C2 and an unknown GSE trimer. The amounts of (+)-catechin and (−)-epicatechin metabolites excreted in urine relative to the quantity of the monomers ingested were 27 and 36 %, respectively, after 24 h. This is similar to the levels of urinary excretion reported to occur by other investigators after feeding (−)-epicatechin to rats and provides further, albeit indirect, evidence that the procyanidin oligomers in the GSE were not depolymerised to monomers to any extent after ingestion. No convincing analytical data were obtained for the presence of flavan-3-ol metabolites in the brain.

The breakdown of glucosinolates, a group of thioglucoside compounds found in cruciferous plants, is catalysed by dietary or microbial myrosinase. This hydrolysis releases a range of breakdown products among which are the isothiocyanates, which have been implicated in the cancer-protective effects of cruciferous vegetables. The respective involvement of plant myrosinase and gut bacterial myrosinase in the conversion, in vivo, of glucosinolates into isothiocyanates was investigated in sixteen Fischer 344 rats. Glucosinolate hydrolysis in gnotobiotic rats harbouring a whole human faecal flora (Flora+) was compared with that in germ-free rats (Flora−). Rats were offered a diet where plant myrosinase was either active (Myro+) or inactive (Myro−). The conversion of prop-2-enyl glucosinolate and benzyl glucosinolate to their related isothiocyanates, allyl isothiocyanate and benzyl isothiocyanate, was estimated using urinary mercapturic acids, which are endproducts of isothiocyanate metabolism. The highest excretion of urinary mercapturic acids was found when only plant myrosinase was active (Flora−, Myro+ treatment). Lower excretion was observed when both plant and microbial myrosinases were active (Flora+, Myro+ treatment). Excretion of urinary mercapturic acids when only microbial myrosinase was active (Flora+, Myro− treatment) was low and comparable with the levels in the absence of myrosinase (Flora−, Myro− treatment). No intact glucosinolates were detected in the faeces of rats from the Flora+ treatments confirming the strong capacity of the microflora to break down glucosinolates. The results confirm that plant myrosinase can catalyse substantial release of isothiocyanates in vivo. The results also suggest that the human microflora may, in some circumstances, reduce the proportion of isothiocyanates available for intestinal absorption.

High energy extruded diets were formulated to contain the same level of protein supplied either by soy protein concentrate (SPC) or fish meal. Three experiments were performed in order to measure voluntary feed intake and feed waste, faecal losses and soluble losses of nitrogen and phosphorus in rainbow trout (average body weight: 100 g). Voluntary feed intake and growth performance of fish fed with demand feeders were not different when diets contained 0, 50 or 75 % SPC instead of fish meal. Total replacement of fish meal by SPC led to a significant decrease in feed intake and resulted in poor growth. This was partly due to methionine deficiency in the SPC based diet. With the addition of crystalline DL-methionine in the diets, an improvement of feed intake and growth performance was apparent. Protein digestibility was high, regardless of the protein source. Excretion of ammonia and urea increased with the level of SPC in the diet. Nitrogen losses decreased when methionine was added to the diet containing only SPC as a protein source. Availability of phosphorus increased with the level of SPC in the diets. Daily soluble losses were not affected by the dietary treatments but the pattern of phosphorus excretion after feed intake was modified. The rise in soluble phosphorus in water occurred later when fish were fed diets with soy protein whatever the dietary level of soy protein concentrate.

A study on nitrogenous and phosphorous waste production in European seabass (Dicentrarchus labrax) was carried out in a flow-through land-based farm. The main objectives were (i) to calculate nitrogenous (N) and phosphorous (P) waste production (particulate and dissolved) from in situ measurements for different rearing ponds with specific fish biomasses, fish sizes and flow rates, (ii) to establish nitrogenous and phosphorous waste production budgets, ratios and equations, and (iii) to compare, for the whole farm, in situ measurements to estimate N and P waste production from waste equations. Waste production was manifest by an increase in concentrations of total ammonia nitrogen, total Kjeldhal nitrogen, particulate nitrogen, dissolved phosphorus and total phosphorus concentrations in the outlet water compared to the inlet water for both batches and whole farm. However, no production of nitrite or nitrate was observed. In our budgets, the N and P amounts from biomass gains and wastes were explained by the N and P derived from feed over the range 83.9−105.2 % and 66.5−104.6 % respectively, depending on the fish batch. Values were respectively 103.4 and 87.5 % for the same calculations in the farm. When the whole-farm waste production was calculated from previous equations derived from batches, and then compared with the measured data, the percentages of recovery (estimated by the ratio predicted data/measured data) were 88 and 94 % for total-N and total-P respectively.

Brown trout and rainbow trout (average weight 100 g) were reared in fresh water at 12 °C under the same conditions before transferring brown trout to sea water, in order to compare phosphorus utilisation in both species. Apparent phosphorus availability, orthophosphate excretion and phosphorus accretion in the fish were directly determined. Thus, actual phosphorus mass balance was built. Rainbow trout raised in fresh water had a higher phosphorus retention coefficient (maximum 50 %) than brown trout reared in fresh water (maximum 45 %). Transferring brown trout to sea water induced a reduction in phosphorus retention (maximum 39 %). Orthophosphate excretion, ranging 7-20 mg phosphorus per kg wet weight per day, represented 10-20 % of ingested phosphorus. Phosphorus availability was lower in brown trout raised in sea water (65 %) than brown trout raised in fresh water (76 %). Phosphorus balance measurements showed that 90 to 98 % of phosphorus flow could be accounted for.

Brown trout and rainbow trout (average weight 100 g) were reared in fresh water at 12 °C under the same conditions before transfer of brown trout to sea water, in order to compare nitrogen utilisation in the two species. Apparent protein digestibility (ADC), nitrogen (ammonia and urea) excretion, protein productive value (PPV) and actual observed nitrogen mass balance were determined. Rainbow trout raised in fresh water had a higher growth rate (1.1 vs 0.8%.d−1), better food conversion ratio (0.7 vs 1.0), better ADC (91 vs 85%) and PPV (45 vs 35%) and lower ammonia excretion rates than brown trout reared in fresh water. Transferring brown trout to sea water induced lower PPV (30%) and ammonia and urea excretion. Salinity did not modify metabolic efficiency in brown trout. Fat content was higher in brown trout (7.7-8.9% ww) than in rainbow trout (5.7-7.6% ww). Nitrogen mass balance indicated that compounds other than ammonia and urea were produced in higher quantities by seawater brown trout. Behaviour, less domestication and specific ability to utilise protein could explain the differences between the two species.

An experiment was carried out to study the effect of changes in either the quality or the quantity of dietary protein intake on the excretion of nitrogen (N) compounds in the chicken. Thirty-two White Rock male broilers (1 day old) were raised in batteries and fed a commercial starter diet for 9 days. Then they were randomly divided into eight groups each of four birds, of similar body weight (mean live weight: 178 (s.e. 1·9) g), and individually housed in metabolism cages. Following a paired-feeding design based on metabolic body weight (kg M0·75), each group of birds was given, for an experimental period of20 days, each of four levels of protein (60, 120, 180 or 240 g/kg: 5 days each) in two groups of isoenergetic (14·5 kj metabolizable energy per g dry matter) and semisynthetic diets based on soya-bean meal, known to be first limiting in methionine, either unsupplemented (diets S) or supplemented with 2 g/kg DL-methionine (diets SM). Excreta were collected every 3 days for 48 h, frozen and stored at −20°C. The excreta samples were subjected to chemical analysis for uric acid by a rapid high-performance liquid chromatographic method, for urea and ammonia by a colorimetric method, and for total N by the Kjeldahl procedure. In general, the excretion of major N compounds was markedly affected by either the quality or the content of dietary protein. Overall, the excretion of total N, uric acid-N, ammonia-N and urea-N significantly (P < 0·05) decreased with improvement in dietary protein quality and significantly (P < 0·05) increased with increase in protein intake. Regression equations were obtained relating the excretion of uric acid, urea, ammonia and total N on protein supply. For the partition ofN compounds output, the ratios of uric acid-N and ammonia-N to total N significantly (P < 0·05) decreased on improving dietary protein quality and increased or remained unchanged, respectively, with the increase in dietary protein content. The use of the ratio of ammonia-N to total N is recommended as a rapid, easy and accurate indicator of dietary protein adequacy, as an alternative to measures based on total N balance, without the need for separation of urine and faeces.

The rates of growth and nitrogen efflux (total nitrogen and ammonia) of individual C. gariepinus ($\bar{x}~=~32.2$ g; S.D. = 4.8 g) kept under 4 feeding regimens, following a 48 h imposed fast (phase l), were measured periodically. In phase 2 (35 d), groups A, B and C were fed a 49.75%, 45.55% or 41.10% protein diet respectively at a ration of 0.5% body weight (1 d−1) Group D were not fed. In phase 3 (25 d) all groups were fed the 41.10% diet. In phase 1, the ammonia efflux rates were lower than any of the values found in either phase 2 and 3. In phase 2, group A (49.75%) had a higher mean ammonia efflux rates than the other groups and ammonia comprised 60-100% of the total nitrogen efflux in all groups. Group D showed a direct relationship between ammonia efflux rate and length of fast. In phase 3, refeeding with the 41.1% protein diet caused the ammonia efflux rates of groups B and D to converge within 7 days to values no different from those of group C, but group A maintained a significantly higher mean value until day 25. During phase 2, the growth rates in B (45.55%) were greatest, but, none of the among groups differences were significant. Group D fish (unfed) lost approximately 30% of their initial weight during phase 2. Nitrogen efflux rates, notably ammonia, showed a pattern of excretion that was directly related to the protein content of the diet, but that the source of dietary protein, dietary energy and the total available energy also influenced nitrogen metabolism. The small differences in growth found were related to between diet differences in composition. The weight loss in the unfed group of fish was probably attributable to the utilisation of lipid and/or protein reserves as metabolic fuels.