Influence of the metabolic state during lactation on milk production in modern sows.

Selection for prolificacy in sows has resulted in higher metabolic demands during lactation. In addition, modern sows have an increased genetic merit for leanness. Consequently, sow metabolism during lactation has changed, possibly affecting milk production and litter weight gain. The aim of this study was to investigate the effect of lactational feed intake on milk production and relations between mobilization of body tissues (adipose tissue or skeletal muscle) and milk production in modern sows with a different lactational feed intake. A total of 36 primiparous sows were used, which were either full-fed (6.5 kg/day) or restricted-fed (3.25 kg/day) during the last 2 weeks of a 24-day lactation. Restricted-fed sows had a lower milk fat percentage at weaning and a lower litter weight gain and estimated milk fat and protein production in the last week of lactation. Next, several relations between sow body condition (loss) and milk production variables were identified. Sow BW, loin muscle depth and backfat depth at parturition were positively related to milk fat production in the last week of lactation. In addition, milk fat production was related to the backfat depth loss while milk protein production was related to the loin muscle depth loss during lactation. Backfat depth and loin muscle depth at parturition were positively related to lactational backfat depth loss or muscle depth loss, respectively. Together, results suggest that sows which have more available resources during lactation, either from a higher amount of body tissues at parturition or from an increased feed intake during lactation, direct more energy toward milk production to support a higher litter weight gain. In addition, results show that the type of milk nutrients that sows produce (i.e. milk fat or milk protein) is highly related to the type of body tissues that are mobilized during lactation. Interestingly, relations between sow body condition and milk production were all independent of feed level during lactation. Sow management strategies to increase milk production and litter growth in modern sows may focus on improving sow body condition at the start of lactation or increasing feed intake during lactation.

Consequences of negative energy balance on follicular development and oocyte quality in primiparous sows†.

Metabolic demands of modern hybrid sows have increased over the years, which increases the chance that sows enter a substantial negative energy balance (NEB) during lactation. This NEB can influence the development of follicles and oocytes that will give rise to the next litter. To study effects of a lactational NEB on follicular development, we used 36 primiparous sows of which 18 were subjected to feed restriction (3.25 kg/day) and 18 were full-fed (6.5 kg/day) during the last 2 weeks of a 24.1 ± 0.3 day lactation. Feed restriction resulted in a 70% larger lactational body weight loss and 76% higher longissimus dorsi depth loss, but similar amounts of backfat loss compared to the full fed sows. These changes were accompanied by lower plasma insulin-like growth factor 1 (IGF1) and higher plasma creatinine levels in the restricted sows from the last week of lactation onward. Ovaries were collected 48 h after weaning. Restricted sows had a lower average size of the 15 largest follicles (-26%) and cumulus-oocyte complexes showed less expansion after 22 h in vitro maturation (-26%). Less zygotes of restricted sows reached the metaphase stage 24 h after in vitro fertilization and showed a higher incidence of polyspermy (+89%). This shows that feed restriction had severe consequences on oocyte developmental competence. Follicular fluid of restricted sows had lower IGF1 (-56%) and steroid levels (e.g., ß-estradiol, progestins, and androgens), which indicated that follicles of restricted sows were less competent to produce steroids and growth factors needed for oocytes to obtain full developmental competence.

Follicular development of sows at weaning in relation to estimated breeding value for within-litter variation in piglet birth weight.

In this study we aimed to identify possible causes of within-litter variation in piglet birth weight (birth weight variation) by studying follicular development of sows at weaning in relation to their estimated breeding value (EBV) for birth weight variation. In total, 29 multiparous sows (parity 3 to 5) were selected on their EBV for birth weight variation (SD in grams; High-EBV: 15.8±1.6, N=14 and Low-EBV: -24.7±1.5, N=15). The two groups of sows had similar litter sizes (15.7 v. 16.9). Within 24 h after parturition, piglets were cross-fostered to ensure 13 suckling piglets per sow. Sows weaned 12.8±1.0 and 12.7±1.0 piglets, respectively, at days 26.1±0.2 of lactation. Blood and ovaries were collected within 2 h after weaning. The right ovary was immediately frozen to assess average follicle size and percentage healthy follicles of the 15 largest follicles. The left ovary was used to assess the percentage morphologically healthy cumulus-oocyte complexes (COCs) of the 15 largest follicles. To assess the metabolic state of the sows, body condition and the circulating metabolic markers insulin, IGF1, non-esterified fatty acid, creatinine, leptin, urea and fibroblast growth factor 21 were analysed at weaning. No significant differences were found in any of the measured follicular or metabolic parameters between High-EBV and Low-EBV. A higher weight loss during lactation was related to a lower percentage healthy COCs (ß= -0.65, P=0.02). Serum creatinine, a marker for protein breakdown, was negatively related to average follicle size (ß= -0.60, P=0.05). Backfat loss during lactation was related to a higher backfat thickness at parturition and to a higher average follicle size (ß=0.36, P<0.001) at weaning. In conclusion, we hypothesise that modern hybrid sows with more backfat at the start of lactation are able to mobilise more energy from backfat during lactation and could thereby spare protein reserves to support follicular development.

Temporary undernutrition during early gestation, corpora lutea morphometrics, ovarian progesterone secretion and embryo survival in gilts.

The present study reports effects of severe undernutrition on luteal function and pregnancy in pigs. Gilts were inseminated and either fasted on Day 10 and 11 after conception (n=11) or fully fed throughout (n=10). Fasting did not affect LH or progesterone pulsatile secretion pattern on Day 11 in samples taken from blood vessels draining an ovary. Ultrasonographic measurements of the size of the corpora lutea did not show any effect of fasting either. However, fasted gilts had 10 to 30% lower systemic progesterone from Day 12 through Day 15 after conception (P<0.05). All gilts farrowed, but fasted gilts had fewer born piglets than fully fed gilts (8.8±0.8 vs 10.9±0.5 respectively; P<0.05). In conclusion, fasting during embryo elongation can compromise embryonic survival by affecting ovarian function in the days after fasting, without having an immediate effect on LH secretion and progesterone output by the ovaries.

Insulin-stimulating diets during the weaning-to-estrus interval do not improve fetal and placental development and uniformity in high-prolific multiparous sows.

Piglet birth weight and litter uniformity are important for piglet survival. Insulin-stimulating sow diets before mating may improve subsequent piglet birth weights and litter uniformity, but the physiological mechanisms involved are not clear. This study evaluated effects of different levels of insulin-stimulating feed components (dextrose plus starch; fed twice daily) during the weaning-to-estrus interval (WEI) on plasma insulin and IGF-1 concentrations, and on follicle development and subsequent luteal, fetal and placental development and uniformity at days 42 to 43 of pregnancy. During WEI, multiparous sows were isocalorically fed diets supplemented with 375 g/day dextrose plus 375 g/day corn starch (INS-H), with 172 g/day dextrose plus 172 g/day corn starch and 144 g/day animal fat (INS-L), or with 263 g/day animal fat (CON). Jugular vein catheters were inserted through the ear vein at 1.5 days before weaning to asses plasma insulin and IGF-1 concentrations. After estrus, all sows received a standard gestation diet until slaughter at days 42 to 43 of pregnancy. The dextrose plus starch-diets enhanced the postprandial insulin response in a dose-dependent manner (e.g. at day 2 insulin area under the curve was 4516 µU/444 min for CON, 8197 µU/444 min for INS-L and 10 894 µU/444 min for INS-H; s.e.m. = 694; P < 0.001), but did not affect plasma IGF-1 concentrations during the first 3 days of WEI. Follicle development and subsequent luteal, fetal and placental development and uniformity were not affected by the dietary treatments, nor related to plasma insulin and IGF-1 concentrations during WEI. Pre-weaning plasma insulin and IGF-1 concentrations were negatively related to sow body condition loss during lactation, but were not related to subsequent reproduction characteristics. This study shows that dietary dextrose plus starch are effective in stimulating insulin secretion (both postprandial peak and long-term concentration), but not IGF-1 secretion during the first 3 days after weaning in multiparous sows. The extreme insulin-stimulating diets during WEI did, however, not improve follicle development, or subsequent development and uniformity of fetuses and placentas in these high-prolific sows (27.0 ± 0.6 ovulations; 18.6 ± 0.6 vital fetuses).

Effects of dietary carbohydrate sources on plasma glucose, insulin and IGF-I levels in multiparous sows.

Effects of different carbohydrate sources on plasma glucose, insulin and insulin-like growth factor-I (IGF-I) levels were compared to subsequently be able to study effects of insulin-stimulating diets on follicle development in sows. The following feed components were tested in 12 sows during six consecutive test periods of 9.5 days: dextrose (DEX), sucrose (SUC), lactose (LAC), dextrose plus lactose (DL), sucrose plus lactose (SL), dextrose plus sugarbeet pulp (DSBP) and control (CON). On day 2, 5 and 9 of each test period, plasma glucose (only at day 9), insulin and IGF-I profiles were determined. Despite similar glucose profiles for all diets, the postprandial insulin response was higher for DL and SL compared with CON and LAC; the other diets were intermediate. Plasma IGF-I levels were higher in CON, LAC and SL compared with DSBP, but differences were only marginal. It was concluded that dextrose and sucrose have the potential to stimulate fast and high insulin peaks, especially when combined with additional lactose. Despite the high dextrose in the DSBP diet, the insulin response was flattened, probably due to the viscosity of sugarbeet pulp. The results show that modulation of plasma insulin levels by dietary carbohydrates seems possible in anabolic sows, but IGF-I levels are less easily modified.

Effects of genetic origin and social environment on behavioral response to manual restraint and monoamine functioning in laying hens.

Purebred laying hen lines of White Leghorn (WL) origin have been found to be more flighty and to show more feather pecking than lines of Rhode Island Red (RIR) origin. It has been found, however, that when RIR birds were housed together with WL birds, RIR birds became more flighty and those mixed groups developed more feather damage than pure-line cage-housed groups. It is unknown, however, whether this effect of social environment is accompanied by changes in stress-related behavior and neurophysiological activity, which are assumed to be associated with increased feather damage. The objective of this study was therefore to investigate the effects of genetic origin (WL or RIR) and social environment (mixed or pure groups) on behavioral response to manual restraint and monoamine functioning. Monoamine functioning was measured by brain serotonin (5-HT) and dopamine turnover. Furthermore, correlations between 5-HT turnover in the brain and peripheral measures of 5-HT in the blood were calculated. Experimental birds, housed either with other birds from the same genetic origin (pure groups) or with both RIR and WL birds (mixed groups) from hatching onward, were subjected to a manual restraint test at 47 wk of age. The WL birds struggled less during restraint and had higher dopamine and 5-HT turnover levels after restraint than did RIR birds. The WL birds also showed higher levels of platelet 5-HT uptake than did RIR birds. No effects of social environment were found. Blood and brain 5-HT measures were found to be correlated, with correlations ranging from 0.34 to 0.57, which seems to offer opportunities for less invasive peripheral indicators of 5-HT activity. In conclusion, genetic origin, but not social environment, affected the behavioral response to manual restraint and monoamine functioning in laying hens.

Postprandial oxidative losses of free and protein-bound amino acids in the diet: interactions and adaptation.

Postprandial oxidation of dietary free amino acids or egg white protein was studied using the [13CO2] breath test in rats, as well as in humans. Thirty-eight male rats were assigned to four dietary test groups. Two diets only differed in their protein fraction. Diet I contained 21% egg white protein. For the breath test egg white protein, intrinsically labelled with [1-13C]-leucine, was used as a substrate. Diet II contained the same amino acids as diet I, though not as egg white protein but in free form. Free [1-13C]-leucine was used to label this diet. In addition, two 1:1 mixtures of both diets were used. During the breath test either the free amino acid or the protein fraction was labelled as in diets I or II. The animals were breath-tested following short-term (day 5) and long-term adaptation (day 20) to their experimental diet. For all diets, including the mixed diets, postprandial oxidative losses on day 5 were significantly higher for the free leucine compared with the protein-derived leucine. Differences between free and protein-derived leucine oxidation had, however, largely disappeared on day 20. The human subjects were breath-tested without any adaptation period to the diets. The oxidative losses of free leucine were also higher than those of protein-derived leucine. None of the studies showed any indication for an interaction between the oxidation of protein-derived amino acids and free amino acids. It is concluded that free and protein-derived amino acids in the diet are mainly metabolized independently.

Dietary amino acids fed in free form or as protein do differently affect amino acid absorption in a rat everted sac model.

In the present study, the effect of free amino acid (FAA) diets on the intestinal absorption rate of methionine and leucine was studied 'ex vivo' with rats adapted for different periods of time to the diets, using the everted sac method. The adaptation period to the 21% FAA diet with an amino acid content based on casein was either, 0 (no adaptation, N-ADA), 5 (short-term adaptation, ST-ADA), or 26-33 days (long-term adaptation, LT-ADA). Within the ST-ADA and the LT-ADA groups, three different levels of methionine were included: 50%, 100% and 200% of the level normally present in casein. All diets were iso-nitrogenous and iso-caloric. After the adaptation period (0, 5, or 26-33 days), intestinal everted sacs were prepared. Methionine or leucine was added to the medium as transport substrate. The methionine absorption rate of the rats of the LT-ADA groups was higher than that of the N-ADA groups. Furthermore, adaptation to 200% dietary methionine levels caused a significantly slower leucine absorption compared to the 100%, and 50% group. Methionine absorption was similar in the 100% and 200% groups, but the absorption of methionine in the 50% group was enhanced in the distal part of the intestines. We concluded that in response diets with 21% FAAs as only amino acid source, amino acid absorption is decreased to avoid toxic effects of high levels of methionine in the circulation.

Dietary amino acids fed in free form and as protein components do not differently affect postprandial plasma insulin, glucagon, growth hormone and corticosterone responses in rats.

This study examined, whether the postprandial fate of dietary amino acids from different amino acid sources is regulated by the responses of insulin, glucagon, corticosterone and growth hormone (GH). Male Wistar rats were cannulated in the vena jugularis and assigned to dietary groups. The diets contained 21% casein or the same amino acids in free form. In the free amino acid diets, methionine level was varied between the groups. The feed was supplied in two distinct meals. In previous experiments it was established that oxidative amino acid losses of the free amino acid diets and protein diets were different. After 3 weeks on those diets, it appeared that the differences in postprandial oxidative losses had been diminished. GH was measured every 12 min, from 144 min before the start of the experimental meal over the following 144 min. Insulin and corticosterone were measured six times from the start of the meal until 270 min after the meal. No differences have been observed between the hormonal responses to both meals at day 5 and at day 26. In conclusion, it has been found that the differences in the oxidative losses between protein and free amino acid meals are not mediated by the combined action of the insulin, glucagon, corticosterone and GH. Postprandial catabolism of amino acids is most probably regulated by substrate induction.

Short-term dynamics in protein and amino acid metabolism.

Actual amounts of free amino acids in the blood are sufficient to support whole body protein synthesis for some minutes only. This indicates that the levels of free amino acids in the circulation are kept small and constant relative to the amounts of amino acids supplied by daily intake and turnover of body proteins. The clearance of the amino acids originating from either endogenous or exogenous sources is mainly due to protein synthesis and metabolic degradation. The partitioning of dietary amino acids between these processes, on the short term, is supposed to play an important role in whole body amino acid economy. Therefore whole body amino acid economy could be improved by nutritional measures that favour the clearance of dietary amino acids by protein synthesis instead of by metabolic degradation. These nutritional measures should to be focused on threshold values for metabolic degradation of individual amino acids.

Effects of acute and chronic level of protein supply on metabolic leucine utilization in growing and mature rats.

Effects of acute (meal) and chronic (diet) level of protein supply on metabolic leucine utilization were investigated in growing (10 weeks) and mature (> 1 year) rats. Rats were conditioned on a high-protein (HP) diet (210 g casein/kg feed) or a low-protein (LP) diet (75 g casein/kg feed) from 7 weeks of age. Overnight-fasted rats were offered a HP or LP meal during a 8 h 14CO2 breath test with a constant infusion of either L-[1-14C]leucine (carboxyl, CL) or L-[U-14C]leucine (universal, UL). Before the meal 14CO2 output was lower for overnight-fasted rats fed on LP than on HP (P < 0.001), and also lower for growing than for mature rats (P < 0.001). Meal ingestion resulted in a rapid increase in 14CO2 output. From 2 h after the start of the meal the effect of acute protein supply on 14CO2 output was significant (P < 0.001), while the effect of chronic protein supply disappeared for CL. After the meal 14CO2 output was transiently lower for growing than for mature rats (P < 0.05), especially after the LP meal. The difference in 14CO2 output between CL and UL increased transiently after the meal, indicating an increase in decarboxylation relative to total oxidation of leucine.(ABSTRACT TRUNCATED AT 250 WORDS)

The metabolic utilization of amino acids: potentials of 14CO2 breath test measurements.

The present paper offers a dual 14CO2 breath test approach to study the metabolic utilization of free amino acids in the body. Using the carboxyl-[14C]isotopomer of an amino acid as the test substrate the percentage recovery of the isotope as 14CO2 reflects which part of the labelled amino acid flux has been decarboxylated. The residual C fragments may flow to total oxidation at least to the level recovered for the universal [14C]isotopomer. In the case that recovery for total oxidation is less than for decarboxylation, part of the [14C]fragments are retained in the body by either exchange or non-oxidative pathways. Utilization of tyrosine and leucine was measured in the post-absorptive phase in adult rats, conditioned on isoenergetic diets containing 210, 75 or 0 g protein/kg. It was shown that the level of dietary protein exerts an influence on both decarboxylation and total oxidation. Although the responses of leucine and tyrosine were not different for total oxidation, there was a difference between the amino acids in their relative rate of decarboxylation. That this dual 14CO2 breath test approach can be used as a tool to evaluate whether the protein and amino acid supply has been adequate to support actual requirements is discussed.

Gel filtration in sodium dodecyl sulphate of hydrophobic muscle proteins on Sephacryl S-400 Superfine.

Gel filtration has been applied to muscle proteins in the presence of the detergent sodium dodecyl sulphate (SDS). This detergent not only solubilizes a variety of proteins but also increases their hydrodynamic volumes, which is a disadvantage for the process of gel filtration. Nevertheless, appropriate choice of gel type, in this case Sephacryl S-400 Superfine, and optimal elution conditions make SDS gel filtration a useful tool in the separation of heterogeneous protein mixtures in the molecular weight range of 20,000-200,000.

A study on the relative turnover of muscle proteins. Comparison of turnover rates in a single step procedure.

In order to enable a direct comparison of the turnover rates of all muscle proteins we introduce a single step procedure. This procedure is based on gel filtration in the presence of a detergent SDS (sodium dodecyl sulphate) which solubilises all muscle proteins. Although the separation is still very crude, some preliminary conclusions may be drawn abut the relative turnover rates of myofibrillar proteins. In our opinion it is unlikely that the myofibril turns over as a unit.