Effect of postweaning feed intake on performance, intestinal morphology, and the probability of diarrhoea in piglets.

Changes in gastrointestinal architecture, high incidence of diarrhoea, and low feed intake (FI) are commonly observed around weaning of pigs, but the relationship between postweaning FI and diarrhoea is unclear. This study aimed to determine the effect of low or high FI during the first days after weaning on growth performance, diarrhoea probability, intestinal permeability, and morphology in pigs until postweaning day (PWD) 28. A total of 120 pigs (7.20 ± 0.26 kg) weaned at 28 days of age (PWD 0) were randomly allocated to five diets and housed individually until PWD 28. Two diets differed in CP and three diets differed in threonine and tryptophan levels. At PWD 4, pigs with the 25% lowest accumulated FI (LOW; n = 30) and 25% highest accumulated FI (HIGH; n = 30) were selected for the study. Faecal consistency was evaluated daily using a 4-scale visual scoring system. Blood was collected at PWD 4, 14, 21 and 28, and small intestinal and colonic tissue was obtained at slaughter on PWD 28. Until PWD 4, LOW pigs consumed approximately 20% (35.7 ± 5.9 g/day) of the FI of HIGH pigs (181 ± 5.75 g/day; P < 0.05) and their average daily gain (ADG) was -103 ± 15.1 g/day. At PWD 28, average daily feed intake, ADG, and feed conversion ratio were still negatively affected by the FI level (P < 0.05) and pigs in the LOW group were on average 4.4 kg lighter than HIGH pigs. Pigs in the HIGH group showed a 55% higher probability of diarrhoea compared with LOW pigs during PWD 0-28. The number of antibiotic treatment days against diarrhoea was 2.38 days higher for HIGH compared with LOW pigs (P = 0.04). The intestinal permeability markers diamine oxidase and D-lactate in plasma were unaffected by the level of FI (P > 0.10). The systemic inflammatory markers haptoglobin and C-reactive protein were higher for HIGH pigs at PWD 4 (P = 0.005), but not affected in the following periods (P > 0.10). Pigs in the HIGH group had an increased area of acidic mucin-producing cells in the small intestine compared with LOW pigs (P < 0.05), but other intestinal morphology measurements at PWD 28 were unaffected by the level of FI. In conclusion, high FI just after weaning was associated with higher growth performance but also higher probability of diarrhoea and more frequent use of antibiotics until PWD 28.

Age-dependent development in protein digestibility and intestinal morphology in weaned pigs fed different protein sources.

Today, weaner diets are optimised using digestibility coefficients obtained from grower-finisher pigs, which may overestimate the digestibility in weaners. The aim of this study was to evaluate the standardised ileal digestibility (SID) of CP and amino acids (AAs), and the intestinal morphology in pigs 0-4 weeks postweaning when fed different protein sources. The experiment included 128 pigs weaned at day 28 and the protein sources were wheat, soybean meal (SBM), enzyme-treated soybean meal (ESBM), hydrothermally treated rapeseed meal (HRSM) and casein. The experiment was conducted as a difference method study including wheat in all diets. Eight pigs were slaughtered on the day of weaning (day 0) and six pigs/treatment were slaughtered at days 7, 14, 21, and 28 postweaning. The SID of CP and AA, as average over the four weeks, was lowest for ESBM and highest for wheat and casein, with SBM and HRSM being intermediate. The SID of CP and AA increased (both linear and quadratic, P < 0.05) over time after weaning. The average SID of CP for all protein sources postweaning was 0.38, 0.59, 0.76, and 0.71 on days 7, 14, 21, and 28, respectively. These differences were significant (P < 0.05) between days 7 and 21, and between days 7 and 28 (P < 0.05), whereas there tended to be a difference between days 7 and 14 (P = 0.06). Protein source did not affect the small intestinal morphology response parameters, whereas time after weaning did. Villous height and villous height to crypt depth ratio differed (P < 0.05) between the days 0 and 7, with shorter villi and a higher ratio at day 7. Crypt depth was not altered between days 0 and 7, or between days 7 and 14. For villi density, crypt density and small intestinal length, a significant increase from days 7 to 14 was observed, but there was no further increase to or difference between days 21 and 28. In conclusion, the low SID of CP in casein on day 7 (0.50) illustrates the challenges related to protein digestion in weanling pigs. The SID of CP and AA is very low during the first two weeks postweaning and time after weaning is more important for protein digestibility, than the source of protein. Fewer mature epithelial cells and less absorptive area in the small intestine in the early postweaning period may partly explain the poor protein digestibility.

Feeding milk supplemented with Ulva sp., Ascophyllum nodosum, or Saccharina latissima to preweaning dairy calves: Effects on growth, gut microbiota, gut histomorphology, and short-chain fatty acids in digesta.

Emerging knowledge shows the importance of preweaning nutrition on programming the gastrointestinal microbiome and development of the gut barrier function. The aim of this study was to assess the effects of supplementing cow milk with either intact dried Ulva sp., Ascophyllum nodosum, or Saccharina latissima on growth performance and several gut health parameters of preweaning dairy calves. Forty male Holstein calves were selected based on birth weight (41 ± 4 kg) and plasma Brix percentage (≥8.7%) at d 2 of life. From d 2 to d 42 of life, the control calves (n = 10) were fed with cow milk (8 L/d) without seaweed supplementation, and the experimental calves were fed with cow milk (8 L/d) supplemented with either Ulva sp. (n = 10), A. nodosum (n = 10), or S. latissima (n = 10) at a concentration of 50 g/8 L of cow milk per day (i.e., 5% on a dry matter basis). Calves were weighed every week, and body weight gain and calf starter intake were monitored weekly. At d 42 ± 3 of life, calves were slaughtered. The organ weights and digesta pH from the reticulorumen, mid- and end small intestine, and mid-colon were recorded. A tissue sample (5 cm) collected from the mid-small intestine was analyzed for histomorphology. Digesta from the mid-small intestine and mid-colon were analyzed for lactobacilli, Escherichia coli, and Enterobacteriaceae, and short-chain fatty acid profile. Weight gain of the calves was not affected by seaweed supplementation. Proportional organ weights were not affected by seaweed supplementation except for reticulorumen weight, which was higher in calves fed Ulva sp. Both the mid-small intestinal and mid-colonic digesta populations of lactobacilli, Enterobacteriaceae, and E. coli, as well as the mid-small intestinal histomorphology in seaweed-supplemented calves were not different from control calves. However, acetic acid proportion in mid-colonic digesta was increased in calves fed Ulva sp. and A. nodosum, whereas butyric acid proportion was decreased compared with the control calves. Digesta pH in mid- and end small intestine and mid-colon were not affected, whereas ruminal pH was increased in calves fed Ulva sp. compared with the control calves. In conclusion, intact dried seaweed supplementation did not improve the growth or selected gut health parameters (i.e., histomorphology, digesta pH, bacteria, and short-chain fatty acids) in preweaning Holstein calves.

Biodegradation of expanded polystyrene by mealworm larvae under different feeding strategies evaluated by metabolic profiling using GC-TOF-MS.

The present study investigated the biodegradation of polystyrene (PS) plastic by mealworm (Tenebrio molitor) on different diets followed by untargeted screening of larvae gut intestine tissue and frass (manure and feed residuals) to investigate the existence of polymer-generated organic residues. Three different diets, consisting of PS, rolled barley and water were tested. PS degradation rates ranged from 16% to 23% within 15 days, with no statistical differences in survival rates. The larvae fed with ad libitum barley:PS (20:1 w/w) and water had the highest growth rate, while higher PS consumption was observed for barley:PS of 4:1 w/w. A GC-TOF-MS analysis revealed no contaminating substances in the gut intestine tissue, nor styrene or PS oligomers, whilst several bioactive compounds and traces of alkanes, mostly with small carbon chains, were present. Metabolomics analysis on the collected frass, either on the lipophilic (CHCl3) or the polar fraction (MeOH-H2O) was performed. Styrene and PS oligomers (dimers, trimers) were identified, though in a relatively low total amount, up to a total of 346.0 ng/mg 2,4 di-tert butylphenol was identified in both frass and tissue, coming from the PS polymer (Non-intentionally added substances; NIAS). Finally, in the polar fraction of frass, bioactive molecules (fatty acids, amides) were identified, together with several hydrocarbons, mostly with longer carbon chains. The formation of these substances indicated enzymatic and biochemical activity in the larvae-gut intestine. It was shown that degrading and contaminating organic compounds occur at low levels, in both gut intestine and frass, during bio-degradation of PS.

Transport of valine across the small intestinal epithelium in pigs fed different valine levels and Bacillus subtilis.

Mutants of Bacillus subtilis overproducing valine (B. subtilis VAL) could be an approach to supply pigs dietary valine (Val). In the study, 18 gilts were fed: (i) negative diet with a standardized ileal digestible (SID) Val:Lys of 0.63:1 (Neg); (ii) Neg added B. subtilis VAL (1.28 × 1011  cfu/kg as-fed) or; (iii) Neg added L-Val to a Val:Lys of 0.69:1. Using the Ussing chamber method, the study aimed to investigate whether (i) the diets affect intestinal transport of additions of 0, 5, 10 or 20 mmol Val/L from the mucosal to the serosal side and (ii) the B. subtilis VAL contributes to a net transport of Val produced in situ. The results showed that the Isc (ΔIscVal ) and release of Val to the serosal side solution (Srel ; µmol cm-2  min-1 ) increased with Val addition (linear and quadratic, p < .0001) but was similar for 5, 10 or 20 mmol Val/L and not affected by diet. No net transport of in situ produced Val by B. subtilis VAL was detected. In conclusion, feeding a Val-deficient diet with or without B. subtilis VAL or a Val sufficient diet did not affect the Val transport across intestinal epithelia. No in situ Val production by B. subtilis VAL was observed in the Ussing chambers.

Evaluation of in situ valine production by Bacillus subtilis in young pigs.

Mutants of Bacillus subtilis can be developed to overproduce Val in vitro. It was hypothesized that addition of Bacillus subtilis mutants to pig diets can be a strategy to supply the animal with Val. The objective was to investigate the effect of Bacillus subtilis mutants on growth performance and blood amino acid (AA) concentrations when fed to piglets. Experiment 1 included 18 pigs (15.0±1.1 kg) fed one of three diets containing either 0.63 or 0.69 standardized ileal digestible (SID) Val : Lys, or 0.63 SID Val : Lys supplemented with a Bacillus subtilis mutant (mutant 1). Blood samples were obtained 0.5 h before feeding and at 1, 2, 3, 4, 5 and 6 h after feeding and analyzed for AAs. In Experiment 2, 80 piglets (9.1±1.1 kg) were fed one of four diets containing 0.63 or 0.67 SID Val : Lys, or 0.63 SID Val : Lys supplemented with another Bacillus subtilis mutant (mutant 2) or its parent wild type. Average daily feed intake, daily weight gain and feed conversion ratio were measured on days 7, 14 and 21. On day 17, blood samples were taken and analyzed for AAs. On days 24 to 26, six pigs from each dietary treatment were fitted with a permanent jugular vein catheter, and blood samples were taken for AA analysis 0.5 h before feeding and at 1, 2, 3, 4, 5 and 6 h after feeding. In experiment 1, Bacillus subtilis mutant 1 tended (P<0.10) to increase the plasma levels of Val at 2 and 3 h post-feeding, but this was not confirmed in Experiment 2. In Experiment 2, Bacillus subtilis mutant 2 and the wild type did not result in a growth performance different from the negative and positive controls. In conclusion, results obtained with the mutant strains of Bacillus subtilis were not better than results obtained with the wild-type strain, and for both strains, the results were not different than the negative control.

Requirement of standardized ileal digestible valine to lysine ratio for 8- to 14-kg pigs.

The objective was to define the Val requirement for weaned piglets in the context of reducing the dietary protein content. A dose-response experiment was conducted to estimate the standardized ileal digestible (SID) Val to Lys ratio required to support the optimum growth of post-weaned piglets. In this study, 96 pigs weighing 8 kg were allotted to one of six dietary treatments (16 pigs for each dietary treatment) and were housed individually. Diets were formulated to provide 0.58, 0.62, 0.66, 0.70, 0.74 and 0.78 SID Val : Lys by adding graded levels of crystalline l-Val to the 0.58 SID Val : Lys diet. Lysine was sub-limiting and supplied 90% of the recommendation (10.95 g SID Lys/kg equal to 11.8 g/kg total Lys). Average daily feed intake (ADFI), average daily gain (ADG) and gain to feed ratio (G : F) were determined during a 14-day period of ad libitum feeding. Blood and urine samples were taken at the end of each week (day 7 and 14 of the experiment) 3 h after feeding the experimental diets. The maximum ADFI and ADG were obtained in pigs fed the 0.78 SID Val : Lys diet; it was not different from the results of pigs fed 0.70 SID Val : Lys diet. The highest G : F was obtained in pigs fed 0.70 SID Val : Lys. The plasma concentration of Val increased linearly (P<0.001) as the dietary SID Val : Lys increased. The increasing dietary Val : Lys also resulted in a linear increase in Cys (P<0.001) and a quadratic increase in Arg (P=0.003), Lys (P=0.05) and Phe (P=0.009). The plasma Gly showed a quadratic decrease (P=0.05) as the dietary Val : Lys increased. Neither plasma nor urinary urea to creatinine ratio was affected by treatment. The minimum SID Val : Lys required to maximize ADFI, ADG and G : F was estimated at 0.67 SID Val : Lys by a broken-line model, and at 0.71 SID Val : Lys by a curvilinear plateau model. The Val deficiency caused a reduction in ADFI, and Val supplementation above the requirement did not impair animal performance. In conclusion, 0.70 SID Val : Lys is suggested as the Val requirement for 8 to 14 kg individually housed pigs.

The optimum ratio of standardized ileal digestible leucine to lysine for 8 to 12 kg female pigs.

The objective of the study was to estimate Leu requirement for weaned piglets to balance indispensable AA in reduced CP diets. A dose-response experiment was conducted to estimate the standardized ileal digestible (SID) Leu to Lys ratio required for the maximum growth of young pigs after weaning. In this study, 96 female pigs (initial BW of 8 kg) were allotted to 1 of 6 dietary treatments with 16 individually penned pigs per treatment. Graded levels of crystalline L-Leu were added to a basal diet to provide diets containing 0.70, 0.80, 0.90, 1.00, 1.10, and 1.20 SID Leu:Lys. Lysine was limiting and fulfilled 90% of the current recommendations. The ADFI, ADG, and G:F were determined during a 2 wk experimental period. Blood and urine samples were taken at the end of each wk. The ADFI increased linearly (P < 0.001) from 0.70 to 0.80 SID Leu:Lys and then remained constant from 0.90 to 1.20 SID Leu:Lys. The ADG showed a quadratic increase ( P= 0.02), as the SID Leu:Lys level increased from 0.70 to 0.90 SID Leu:Lys and did not change further from 0.90 to 1.20 SID Leu:Lys. The G:F increased quadratically (P < 0.001) with increasing SID Leu:Lys level, and the greatest G:F was achieved with pigs receiving the diet with 0.80 SID Leu:Lys. Increasing the dietary SID Leu:Lys resulted in a linear increase in plasma Leu concentration (P < 0.001) and quadratic increases (P < 0.001) in plasma Cys concentration. The plasma concentration of most of the other AA was lowest in pigs receiving the diets with 0.90 to 1.00 SID Leu:Lys. The plasma urea nitrogen concentration tended (P = 0.08) to be lowest in pigs receiving 1.00 SID Leu:Lys, suggesting a more balanced AA profile at this level. Using a curvilinear-plateau model, the SID Leu:Lys requirement was estimated at 0.93 to maximize growth in female pigs weighing 8 to 12 kg.

Isoleucine requirement of pigs weighing 8 to 18 kg fed blood cell-free diets.

The aim of the present study was to determine the minimum requirement of Ile in young pigs, enabling feeding of balanced low-CP diets. Most previous studies have used experimental diets that included blood cells, which are particularly high in Leu and known to antagonize the use of Ile. One week after weaning at d 28, 100 crossbred female pigs weighing 7.9 ± 0.7 kg were allocated to 1 of 5 dietary treatments. Diets were formulated to contain 1.15 g standardized ileal digestible (SID) Lys/MJ NE and were free of blood cells. The SID Ile was 0.42, 0.47, 0.53, 0.58, and 0.62 relative to Lys. The other indispensable AA were supplied according to requirements. Representative samples from the 5 diets were analyzed in 4 replicates at 3 different laboratories. The pigs were fed ad libitum and individually housed in 7 identical rooms during a 21-d period. At d 0, 7, 14, and 21, the pigs were weighed, and feed intake was determined. At d 15, blood samples were collected from the jugular vein to determine the plasma urea and free AA content. There were differences among the 3 laboratories in the analyzed content of several AA, and also Ile and Lys showed a large variation within the diets, which may cause variation in published requirement estimates. The concentration of Ile in plasma increased linearly (P < 0.01), and Lys in plasma decreased linearly (P = 0.02) with increasing SID Ile:Lys. A tendency for a linear decrease in plasma concentration was found for Thr (P = 0.10). Both ADFI and ADG were reduced when Ile was supplied above the Ile requirement estimate. Quadratic regression curves on ADFI, ADG, and G:F all showed the maximum at 0.52 SID Ile:Lys. Modeling with 2-sloped quadratic broken-line curves showed the maximum at 0.50, 0.53, and 0.54 SID Ile:Lys for ADFI, ADG, and G:F, respectively. In conclusion, the average estimation of requirement in this dose-response study using blood cell-free diets was 0.52 SID Ile:Lys during a 21-d experimental period from 8 kg BW.

Ileal digestibility of sunflower meal, pea, rapeseed cake, and lupine in pigs.

The standardized ileal digestibility (SID) of CP and AA was evaluated in soybean (Glycine max) meal, sunflower (Helianthus annuus) meal, rapeseed cake, and field pea (Pisum sativum) using 10 pigs and in lupine (Lupinus angustifolius) using 7 pigs. Pigs were fitted with either a T-cannula or a steered ileocecal valve-cannula. Diets contained 170 to 186 g CP/kg DM. Endogenous losses of CP and AA were estimated by feeding a N-free diet. The SID was calculated using the average of Cr(2)O(3) and TiO(2) as indigestible markers and corrected for type of cannula. The SID of CP was greater (P < 0.05) for soybean meal and pea compared to sunflower meal, rapeseed cake, and lupine. The SID of Lys and His were lowest (P < 0.05) in sunflower meal, and the SID of Met and Val were lowest (P < 0.05) in lupine. These results imply soybean meal and pea to be a high-digestible protein source relative to sunflower meal, rapeseed cake, and especially lupine, although all tested feedstuffs seem appropriate for inclusion in diets for organic pigs.

Effect of microbial phytase on phosphorus digestibility in non-heat-treated and heat-treated wheat-barley pig diets.

The objective was to evaluate effects of microbial phytase on apparent total tract digestibility (ATTD) of P in a non-heat-treated and a heat-treated wheat (Triticum aestivum)-barley (Hordeum vulgare) diet fed without inorganic P in a 2 × 3 factorial arrangement. The basal diet was ground and half of the batch was steam pelleted at 81°C and crumbled. Phytase was added at 0, 250, and 500 phytase units (FTU)/kg as-fed (Aspergillus niger). The study comprised 36 pigs from 6 litters. Pigs were housed in metabolism crates and fed 1 of 6 diets for 12 d: 5 d for adaptation and 7 d for total collection of feces. The ATTD of P was highest (P < 0.01) for the non-heat-treated diets and highest (P < 0.01) for the phytase-supplemented diets. Heat treatment reduced plant phytase activity by 25% whereby the ATTD of P decreased (P < 0.01) from 57 to 49%. Microbial phytase increased the ATTD of P to a maximum of 64 and 61% in the non-heat-treated and heat-treated diets corresponding to an increase of 7 and 12%-units. Responses for ATTD of P did not differ between 250 vs. 500 FTU/kg as-fed. In conclusion, processing of feed (meal or pellets) containing plant phytase should be considered to avoid over- or underestimation of effects of microbial phytase.

High-moisture air-tight storage of barley and wheat improves nutrient digestibility.

Barley (Hordeum vulgare) and wheat (Triticum aestivum) are often stored dry with 14% or less moisture, which during rainy periods may require that grains are dried after harvest. The hypothesis is that air-tight storage of high-moisture barley and wheat will increase nutrient digestibility due to chemical conversions prior to feeding. The objective was to evaluate the effect of high moisture compared to dry storage of barley and wheat on digestibility of P and CP. The crops were grown on 1 field keeping other factors constant. Half of the grains was harvested in the morning after a rainy day and stored in air-tight silos (DM, %: barley, 85.2; wheat, 82.8) and the other half was harvested later the same day (windy and sunny) and stored dry (DM, %: barley, 89.8; wheat, 88.3). After 6 mo of storage, 1 low- and 1 high-moisture diet were prepared with a barley:wheat ratio of 1:1 mixed with soybean (Glycine max) meal and rapeseed cake to produce a compound diet without inorganic P and microbial phytase. Sixteen 45-kg pigs housed in metabolism crates were fed either the low- or the high-moisture diet for 5 d for adaptation and 7 d for total collection of feces. Digestibility of P was 12% higher (P < 0.01) and of CP was 4% higher (P = 0.08) in the high-moisture diet. Phytase activity of dry-stored grain was lower (P < 0.01) and phytate P was 4% higher in the high-moisture stored grain vs. the grains stored dry. Overall, high-moisture storage increased digestibility of P and CP when the grain was fed to finishing pigs. Therefore, high-moisture air-tight storage saved energy (without drying) and at the same time enhanced P digestibility and increased the nutritional value of grain probably through enzymatic activity during storage.

Mammary remodeling in primiparous and multiparous dairy goats during lactation.

Milk production is generally lower but lactation persistency higher in primiparous (PP) than in multiparous (MP) goats. This may be related to differences in development and maintenance of mammary gland function, but the underlying mechanisms are not well understood. The present study aimed to elucidate whether differences in lactational performance between PP and MP mammary glands are related to the time course of development and maintenance, not only of the mammary epithelial cell (MEC) population, but also of the mammary vasculature that sustains synthetic activity. Mammary biopsies were obtained from both mammary glands of 3 PP and 6 MP (>or=2 parity) dairy goats at parturition (d 1), d 10, 60, and 180 of lactation. Gene transcription relating to MEC turnover and vascular function was quantified by real-time reverse transcription-PCR, mammary morphology was characterized (quantitative histology), and cell turnover was determined (terminal deoxynucleotidyl transferase dUTP nick end labeling assay and Ki-67). Primiparous glands showed higher expression for the genes involved in angiogenesis; namely, vascular endothelial growth factor receptor 2, and angiopoietin 1 and 2 and their receptor, a few days after parturition (d 10). Primiparous glands also had higher rates of MEC proliferation in early lactation. It therefore appears that initiation of lactation is associated with development and growth of the mammary gland into early lactation, which continues for a longer period in PP compared with MP glands. In addition, MEC survival was found to be higher in PP glands throughout lactation, and MEC in PP glands underwent more extensive differentiation. This could explain the reported flatter lactation curve and higher lactation persistency in PP glands. Although some of the genes included in this study were differentially expressed in PP and MP glands during the course of lactation, it was not possible to identify any specific genomic factor(s) that could account for the differences between PP and MP glands with respect to mammary development and MEC survival during lactation. It remains to be established why parity number affects MEC and vascular development and survival during lactation, and, in particular, which regulatory mechanisms are involved.

Continuous lactation effects on mammary remodeling during late gestation and lactation in dairy goats.

The present study aimed to 1) elucidate whether continuous milking during late gestation in dairy goats negatively affects mammary remodeling and hence milk production in the subsequent lactation, and 2) identify the regulatory factors responsible for changes in cell turnover and angiogenesis in the continuously lactating mammary gland. Nine multiparous dairy goats were used. One udder half was dried off approximately 9 wk prepartum (normal lactation; NL), and the other udder half of the same goat was milked continuously (continuous lactation; CL) until parturition or until the half-udder milk yields had dropped to below 50 g/d. Mammary biopsies were obtained from each udder half just before the NL gland was dried off (before dry period), within the first 2 wk after drying-off (early dry period, samples available only for NL glands), in the mid dry period, within the last 2 wk before parturition (late dry period), and at d 1 (the day of parturition), 3, 10, 60, and 180 of lactation. Mammary morphology was characterized in biopsies by quantitative histology, and cell turnover was determined by immunohistochemistry (terminal deoxynucleotidyl transferase dUTP nick end labeling and Ki-67). Transcription of genes encoding factors involved in mammary epithelial cell (MEC) turnover and vascular function was quantified by quantitative reverse transcription PCR. Results demonstrated that omitting the dry period was possible in goats but was not as easy as claimed before. Renewal of MEC was suppressed in CL glands, which resulted in a smaller MEC population in the subsequent lactation. At the time of parturition (and throughout lactation), the mammary glands subjected to CL had smaller alveoli, more fully differentiated MEC, and a substantially larger capillary fraction compared with NL glands. The continuously lactating gland thus resembled a normally lactating gland in an advanced stage of lactation. None of the studied genomic factors could account for these treatment differences. The described characteristics in CL glands compared with NL glands indicated a gland maintained in lactation for a longer period.

Absorption and metabolism of benzoic acid in growing pigs.

Dietary benzoic acid (BA) supplementation causes a pronounced reduction in urinary pH but only small changes in blood pH. The present study aimed to investigate the portal absorption profile, hepatic metabolism of BA, and renal excretion of hippuric acid (HA) underlying the relatively small impact of BA on systemic acid-base status. Eight growing pigs (BW = 63 +/- 1 kg at sampling) fitted with permanent indwelling catheters in the abdominal aorta, hepatic portal vein, hepatic vein, and mesenteric vein were allocated to 4 sampling blocks and randomly assigned to control (CON; nonsupplemented diet) or BA supplementation (B; control diet + 1% BA top-dressed). Feed intake was restricted to 3.6% of BW and the ration divided into 3 equally sized meals offered at 8-h intervals. Blood pH (7.465 and 7.486 +/- 0.004) and urinary pH (4.99 and 7.01 +/- 0.09) were less (P = 0.03 and P < 0.01) in B compared with CON. The arterial concentration, net portal flux, and net hepatic uptake of BA increased (P < 0.01) in B compared with CON. The net portal flux of BA increased (P < 0.01) after feeding with B, but remained positive (P < 0.01) at all sampling times (n = 8). Recovery of dietary BA as increased net portal flux and hepatic uptake of BA was 87 +/- 5% and 89 +/- 15%, respectively. The recovery of dietary BA as urinary excretion of BA and HA was 0.08 +/- 0.02% and 85 +/- 7%, respectively. It is concluded that the small impact of BA supplementation on systemic acid-base status was caused by a protracted BA absorption and efficient hepatic extraction and glycine conjugation in combination with efficient renal clearance of HA. Together, these physiological mechanisms prevented major BA and HA accumulation in body fluids.

Cellular mechanisms in regulating mammary cell turnover during lactation and dry period in dairy cows.

The mechanisms involved in regulating mammary cell turnover during the pregnancy-lactation cycle in dairy cows are unclear. The objective of present experiment was to describe expression of genes encoding proteins known to be involved in pathways regulating mammary cell proliferation, apoptosis, differentiation, cell survival, and tissue remodeling. Mammary gland biopsies were taken 7 times during the pregnancy-lactation cycle of 10 dairy cows, and samples were analyzed by immunohistochemistry and real-time PCR. Cell proliferation was greatest during the dry period and apoptosis was high in early dry period and early lactation. Based on Fas (tumor necrosis factor receptor superfamily member 6), Fas ligand, and caspase-3, caspase-8, and caspase-9 gene expression, no indication was found of a stage-dependent shift between the extrinsic and intrinsic pathways leading to apoptosis. Gene expression of microsomal glutathione S-transferase (mGST) did not vary significantly, whereas B-cell leukemia/lymphoma 2 (Bcl-2) and BCL2-associated X protein (Bax) gene expression was greatest during the dry period and early lactation and coincided with high cell turnover. Gene expression of early response genes c-Fos, c-Jun, and c-Myc correlated to neither rate of cell proliferation nor plasma concentration of insulin-like growth factor (IGF)-I and insulin. Gene expression of nuclear factor of kappa light chain gene enhancer in B-cells (NFkappaB) and NFkappaB inhibitor alpha was greatest in the periparturient period, and NFkappaB gene expression coincided with an anticipated need for cell survival factors. Expression of transforming growth factor beta (TGF-beta) receptor 1 and 2 mRNA was greatest in early lactation, whereas TGF-beta1 did not vary significant during the pregnancy-lactation cycle. Even though our results on the TGF-beta system did not comply with other studies, the gene expression pattern of the TGF-beta receptors indicates a role in regulating apoptosis in early lactation. Signal transducer and activator of transcription 5 (STAT5) gene expression was high in the periparturient period, which suggests a role for STAT5 in regulation of mammary cell proliferation and differentiation in dairy cows. Expression of tissue-plasminogen activator, plasminogen activator inhibitor-1, and IGF binding protein 5 genes was greatest in early lactation, suggesting a role for IGF binding protein 5 in coordinating regulation of apoptosis and tissue remodeling.

Effect of pregnancy and feeding level on cell turnover and expression of related genes in the mammary tissue of lactating dairy cows.

Milk yield is reduced by pregnancy, and the present experiment was conducted to study the biological basis for the negative effect of pregnancy on milk yield. A total of 16 dairy cows were fed at either a normal or a low feeding level (eight cows per treatment), and half of them were inseminated after approximately 3 months of lactation and the other half were not inseminated. Mammary biopsies were taken at approximately 9 months of lactation. The milk yield of pregnant cows was reduced by 2.6 kg/day, and lactation persistency was reduced already from the time of insemination. Low feeding level reduced the milk yield by 9.8 kg/day from week 8 to week 39 of lactation, whereas no interaction between pregnancy and feeding level was found. Cell proliferation (Ki-67) and apoptosis (terminal deoxynucleotidyl transferase dUTP nick end labeling, TUNEL) were unaffected by feeding level, and pregnancy tended to reduce cell proliferation but had no effect on apoptosis. Reduced cell proliferation may explain the reduced lactation persistency in pregnant cows. Transcription of oestrogen receptor α, progesterone receptor A and B, and long and short isoforms of the prolactin receptor were higher in pregnant cows compared with non-pregnant cows. Feeding level did not mediate changes in transcription of genes. Transcription of other cell-turnover-related genes (IGF-I, IGF binding protein-5, caspase-3) as well as genes related to the secretory activity of the cells (α-lactalbumin and acetyl CoA carboxylase α) was not affected by pregnancy or by feeding level.

Cell turnover and activity in mammary tissue during lactation and the dry period in dairy cows.

Milk yield of the dairy cow follows a pattern termed the lactation curve. We have investigated the cellular background for this pattern. Seven mammary biopsies were obtained from each of 10 cows: at the end of lactation (d 347, equal to d 77 before next parturition); during the dry period at d 48 (4 d after dry off); 16 d before parturition; and during lactation at d 14, 42, 88, and 172. The fraction of proliferating (staining positive for Ki-67) alveolar cells was higher during the dry period (8.6%) than during lactation (0.5%). The fraction of apoptotic (staining positive by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling) alveolar cells was higher immediately after dry off (0.37%) and in early lactation (0.76%) than during other periods (0.15%). The enzyme activities of fatty acid synthetase, acetyl CoA-carboxylase, and galactosyl transferase were approximately 12-, 11-, and 4-fold higher, respectively, during lactation than during the dry period. In conclusion, mammary cell proliferation is substantial in a period near parturition but otherwise low, and apoptosis is elevated at dry off and in early lactation. The increase in apoptosis in early lactation may be due to discarding nonfunctional or senescent cells or to removal of a surplus of newly synthesized cells. The activity of selected enzymes central for milk synthesis is probably not limiting for milk production.

Inter-laboratory validation study of five commercial ELISA test kits for the determination of peanut proteins in biscuits and dark chocolate.

The results of an inter-laboratory study with five commercially available peanut ELISA test kits to detect and quantify peanut residues in two food matrices (biscuit and dark chocolate) at four different concentrations (0-10 mg peanut kg(-1) matrix corresponding to about 0-2.5 mg peanut protein kg(-1) matrix) are reported. In general the five ELISA test kits evaluated could detect peanut protein in the two food matrices. In three cases, the study challenged the test kits beyond their intended use for quantification below the manufacturers' defined cut-off limits. Generally, all five ELISA test kits performed well in the concentration range 5-10 mg kg(-1) rather than in the low concentration range (2.0 or 2.5 mg kg(-1)). The variation in the found recoveries of peanut between the different test kits had a spread of 44-191% across all concentrations. The quantification characteristics between test kits differed significantly at the very low mg kg(-1) level. Two test kits performed well even at concentrations below 5 mg kg(-1) with reproducibilities of 27-36% for biscuits and 45-57% for chocolate.

The International Measurement Evaluation Programme, IMEP-8: carbon and oxygen isotope ratios in CO(2).

In IMEP-8, two samples of high purity CO(2)(g), with different carbon and oxygen isotope ratios were distributed to 27 participants, originating from 14 countries and from various isotopic measurement domains (geochemistry, atmospheric and food chemistry), but particularly set up for food laboratories. In total 19 laboratories reported results. The outcome of this comparison exercise shows that the laboratories reported carbon and oxygen isotope results in good agreement with the reference values across the domains. The reported results for delta(13)C(VPDB) (carbon) for both materials are within 1 per thousand. However, for the reported results of delta(18)O(VPDB) (oxygen) for both materials the overall spread of the reported results is about 11 per thousand. Within this spread two distinct groups of participants can be identified, where the results within each group vary about 2 per thousand. The latter seems to be caused by calculation errors by participants of the reporting delta(18)O(VPDB) values. As requested, participants also reported the isotope amount ratio for carbon and oxygen in the CO(2) samples. For carbon, all reported results for both materials agree with the isotope ratio value, which can be traced back to the value reported by Craig. For oxygen, all results are in good agreement and deviate by a maximum of 0.5% from the reference values measured at IRMM. Work carried out indicates the carbon isotope ratio, for both samples IMEP-8A and IMEP-8B, differ from those reported by Craig by as much as 1.2%. In the case of oxygen, this deviation is far smaller. Both data sets, i.e. the one realised by Craig and the one realised at IRMM, demonstrate traceability to SI. It is clear that both values significantly disagree.