Effect of dietary mineral phosphorus and phytate on in situ ruminal phytate disappearance from different concentrates in dairy cows.

The first objective of this study was to determine the influence of dietary composition on the in situ disappearance of phytate (InsP6) from wheat, corn, soybean meal, and rapeseed meal [solvent-extracted, without (RSM) or with (hRSM) heat treatment] in the rumen of dairy cows. The second objective was to assess the primary degradation products of InsP6 in the rumen. Three diets differing in phosphorus and InsP6 concentration (basal diet = 0.38% P in dry matter; high-P diet = 0.56% P; high-InsP6 diet = 0.39% P) were fed to 3 ruminally fistulated lactating Jersey cows in a 3 × 3 Latin square. Ground concentrates (sieve size = 2 mm) were incubated in polyester bags in the rumen for 2, 4, 8, 16, and 24 h. The bag residues were analyzed for P, InsP6, isomers of lower inositol phosphates (InsP5, InsP4, InsP3), and crude protein. The InsP6 disappeared more rapidly from cereal grains than from oilseed meals; however, after 24 h of incubation ≥95% InsP6 had disappeared from all concentrates except hRSM (57%; diet average). Feeding the high-InsP6 diet increased InsP6 disappearance for oilseed meals, but not for corn and wheat. The predominant InsP5 isomer in all bag residues was Ins(1,2,4,5,6)P5 followed by Ins(1,2,3,4,5)P5 and Ins(1,2,3,4,6)P5. A further InsP5 isomer [Ins(1,3,4,5,6)P5] was detected in both rapeseed meal bag residues. Feeding the high-InsP6 diet led to lower concentrations of Ins(1,2,4,5,6)P5 and Ins(1,2,3,4,5)P5, whereas an interaction between diet, concentrate, and time occurred for Ins(1,2,3,4,6)P5 and Ins(1,3,4,5,6)P5. The results confirm the high potential of rumen microorganisms to hydrolyze InsP6; however, increasing the amount of InsP6 in the diet can further enhance InsP6 hydrolysis, which may be relevant when concentrates with slowly degradable InsP6, such as RSM or heat-treated concentrates, are fed to dairy cows. Based on the concentrations of InsP5 isomers, 3 and 6 phytases appear to play a major role in the rumen. Conversely, intrinsic plant phytase activity appears to be less relevant as the percentage of its primary hydrolysis product, Ins(1,2,3,4,5)P5, changed only slightly upon using wheat known for high intrinsic phytase activity instead of the other concentrates. Additional information regarding the factors influencing the extent of ruminal InsP6 disappearance will require further studies to determine the phytase activity of rumen microorganisms and the characteristics of their respective phytases.

Ruminal phytate degradation of maize grain and rapeseed meal in vitro and as affected by phytate content in donor animal diets and inorganic phosphorus in the buffer.

The ruminal disappearance of phytate phosphorus (InsP6 -P) from maize grain and rapeseed meal (RSM) was determined in two in vitro studies. In experiment 1, two diets differing in phosphorus (P) and InsP6 -P concentration were fed to the donor animals of rumen fluid (diet HP: 0.49% P in dry matter, diet LP: 0.29% P). Maize grain and RSM were incubated in a rumen fluid/saliva mixture for 3, 6, 12 and 24 h. In experiment 2, a diet similar to diet HP was fed, and the rumen fluid was mixed with artificial saliva containing 120 mg inorganic P/l (Pi) or no inorganic P (P0). Maize grain and RSM were incubated with either buffer for 3, 6, 12 and 24 h. Total P (tP) and InsP6 concentration were analysed in the fermenter fluids and feed residues. The disappearance of InsP6 -P from maize was completed after 12 h of incubation in both experiments. From RSM, 93% (diet LP) and 99% (diet HP) of the InsP6 -P in experiment 1 and 80% (Pi) and 89% (P0) in experiment 2 had disappeared after 24 h of incubation. InsP6 -P disappearance was higher when diet HP was fed (maize: 3 and 6 h; RSM: 6 and 24 h of incubation) and when rumen fluid was mixed with buffer P0 (maize: 6 h; RSM: 12 and 24 h of incubation). InsP6 -P concentration in the fermenter fluids was higher for maize, but no accumulation of InsP6 -P occurred, indicating a prompt degradation of soluble InsP6 . These results confirmed the capability of rumen micro-organisms to efficiently degrade InsP6 . However, differences between the feedstuffs and diet composition as well as the presence of inorganic P in the in vitro system influenced the degradation process. Further studies are required to understand how these factors affect InsP6 degradation and their respective relevance in vivo.

Molecular marker assisted broadening of the Central European heterotic groups in rye with Eastern European germplasm.

Broadening the genetic base of heterotic pools is a key to ensure continued genetic gains in hybrid breeding and extend hybrid cultivation to new areas. In the present study, two Central European heterotic pools (Carsten and Petkus) and five Eastern European open-pollinated varieties (OPVs, Pop-1 to Pop-5) were studied with the objectives to (1) investigate the genetic diversity in OPVs and the heterotic pools using molecular and field data, (2) evaluate the molecular diversity among OPVs, (3) examine the combining ability for grain yield of the OPVs when crossed with testers in field trials, and (4) develop a strategy for targeted introgression of OPV germplasm into the heterotic pools. In total, 610 S(0) plants, 347 from OPVs and 263 from heterotic pools, were developed. Clones of the S(0) plants of OPVs were crossed with two testers belonging to each heterotic pool, while clones of heterotic pools were crossed with only the opposite tester. Testcrosses were evaluated for grain yield in multi-location trials. In addition, 589 S(0) plants were fingerprinted with 30 SSR markers. The data revealed that the Carsten pool has a narrow genetic base and should be the primary target for broadening the established heterotic pattern. Mean and genetic variance suggested that Pop-2 and Pop-4 are good candidates for introgression in Petkus pool and Pop-5 in Carsten pool. Nevertheless, introgression of Pop-5 in Carsten could reduce the genetic diversity between heterotic pools. Therefore, we suggest that either selected plants of Pop-5 should be introgressed or more Eastern European germplasm should be fingerprinted and field evaluated to identify promising germplasm for broadening the established heterotic pattern.

Testcross performance of rye introgression lines developed by marker-assisted backcrossing using an Iranian accession as donor.

Introgression libraries facilitate the identification of favorable exotic alleles or genomic regions, which can be exploited for improving elite breeding material. We evaluated the first two introgression libraries in rye (Secale cereale L.) on the phenotypic and molecular level. Our objectives were to detect candidate introgression lines (pre-ILs) with a better testcross performance than the recurrent parent and identify donor chromosome segments (DCS) responsible for the improved performance. We introduced DCS from the self-incompatible heterozygous exotic Iranian primitive rye accession Altevogt 14160 (donor) into the genetic background of the elite inbred line L2053-N (recurrent parent) by marker-assisted backcrossing and developed 40 BC(2)S(3) lines in each introgression library. Testcross performance for three agronomic and six quality traits was evaluated in replicated field trials across two testers at five locations over 2 years. The phenotypic effect of the DCS was analyzed for all traits. The pre-ILs had on average a testcross performance comparable to that of the recurrent parent. Significant (P < 0.05) differences between individual pre-ILs and the recurrent parent were detected for all traits except for heading date. For more than 60% of the significant (P < 0.05) differences, the pre-ILs were superior to the recurrent parent. For some pre-ILs, specific DCS were identified containing presumably quantitative trait loci responsible for the superior hybrid performance. Consequently, our study revealed that the development and employment of introgression libraries offers the opportunity for a targeted increase of genetic diversity of elite rye material for hybrid performance of agronomically important traits.

Phytate degradation and phosphorus digestibility in broilers and turkeys fed different corn sources with or without added phytase.

The aim of the present study was to test whether different dietary corn sources and phytase supplementation affect the prececal phosphorus digestibility (pcdP) and appearance of inositol phosphates in the lower ileum of growing broiler chickens and turkeys. Two experiments were conducted, one with broiler chickens and one with turkeys. Four corn diets were provided; these were formulated to contain low P and calcium (Ca) contents and incorporated 43% of one of the four different corn sources. Diets were either unsupplemented or supplemented with 500 FTU of an Escherichia coli-derived phytase/kg feed. Experimental diets were fed ad libitum from day 20 post-hatch. At 28 d of age, digesta were sampled from the lower ileum of animals to determine pcdP and pc myo-inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate) (InsP6) degradation and to analyze the concentrations of lower inositol phosphate isomers. The pcdP of non-supplemented diets ranged from 51 to 60% and from 22 to 28% in broilers and turkeys, respectively. A negative correlation was observed between the InsP6 content of the corn source and the pcdP of diets in broilers only. Without phytase supplementation, pc InsP6 degradation ranged from 64 to 76% in broilers and from 6 to 15% in turkeys. Phytase increased the pcdP by around 15% in broilers (P < 0.001) and 9 to 17% in turkeys (P < 0.001). In turkeys, phytase efficacy was greatest when the diets contained corn with higher contents of ether extract and InsP6. An effect of corn source on the appearance of lower InsPs in the ileal digesta was found in broilers only. These results suggest that broilers possess a greater capacity for InsP6 degradation and hydrolysis of lower InsPs compared with turkeys. Furthermore, the results are influenced by the corn source used. Further research is needed to identify the factors responsible for the low level of phytate degradation in turkeys in order to improve the availability of InsP6-P and the efficacy of phytase.

Variability of prececal phosphorus digestibility of triticale and wheat in broiler chickens.

The aim of this study was to evaluate the P digestibility of different wheat and triticale genotypes in growing broiler chickens. Additionally, the relationships between physical or chemical properties of genotypes and P digestibility were determined. A low P, low Ca basal diet based on cornstarch and soybean meal was supplemented with 20% or 40% of 8 different wheat or triticale genotypes at the expense of cornstarch. Experimental diets were fed to broilers between d 20 and 26 of age before digesta samples were collected from their lower ileum for determination of prececal P digestibility (pcdP). Triticale-based diets had an average pcdP of 54%. Neither the concentration of triticale nor the genotype itself affected the pcdP of diets. The pcdP of triticale genotypes calculated by linear regression analysis ranged from 53% to 78%. No correlations were found between physical or chemical properties (viscosity, phytase activity, total and phytate P contents) and the pcdP of triticale genotypes. In contrast, a significant (P < 0.05) effect of genotype and wheat concentration was observed in wheat-based diets. The average pcdP of diets decreased from 60% to 50% by increasing the wheat concentration from 20% to 40%. As no linear relationship was observed between P intake and the amount of pcdP in the diet, the pcdP (%) of wheat genotypes was calculated separately for both concentrations, and accounted for 38% to 67% and 20% to 38% with 20% and 40% wheat inclusion in diets, respectively. Single chemical and physical characteristics could not explain the observed differences in P digestibility. Our results revealed a high variation in the pcdP of different wheat and triticale genotypes that should be considered in diet formulation. However, further research is needed to identify factors that contribute to variation of P digestibility in both grains.

Effect of supplemental phytase and xylanase in wheat-based diets on prececal phosphorus digestibility and phytate degradation in young turkeys.

This study aimed to investigate the effect of phytase and a combination of phytase and xylanase on the prececal phosphorus digestibility (pcdP) of wheat-based diets in turkeys. A low-P basal diet (BD) based on cornstarch and soybean meal, and 2 diets containing 43% of different wheat genotypes (genotype diets GD6 or GD7) were fed to turkeys from 20 to 27 d of age. Diets were fed either without enzyme supplementation or supplemented with phytase (500 FTU/kg) or a combination of phytase and xylanase (16,000 BXU/kg). At 27 d of age, digesta were sampled from the lower ileum of animals to determine pcdP and pc myo-inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate) (InsP6) disappearance, and to analyze the concentrations of lower inositol phosphate isomers. Similar pcdP was observed in non-supplemented BD and GD (∼36%). Phytase alone increased the pcdP in all diets by 8 to 12%, but a beneficial effect of xylanase was found only for BD. Similar results were found for pc InsP6 disappearance, although xylanase addition compared to phytase alone decreased pc InsP6 disappearance in GD7 compared to phytase alone. Animals fed GD7 performed better than those fed GD6; however, these differences could not be linked to the pcdP. The pattern of lower inositol phosphates in digesta also changed with enzyme supplementation, resulting in lower proportions of InsP5 and higher proportions of InsP4. Phytase alone decreased Ins(1,2,3,4,6)P5 but increased D-Ins(1,2,3,4,5)P5 and D-Ins(1,2,5,6)P4 concentrations. An additional increase in D-Ins(1,2,3,4,5)P5 and D-Ins(1,2,5,6)P4 concentrations was achieved with xylanase, although for the former isomer, this was observed only with GD. These results indicate that enzyme supplementation alters the pc degradation of InsP6, and that combining both enzymes had a minor additional effect on the pcdP from wheat-based diets when compared to phytase alone.

Variability in amino acid digestibility of triticale grain from diverse genotypes as studied in cecectomized laying hens.

Triticale, an anthropogenic hybrid grain, is increasing in importance as a feed grain for laying hens. However, our limited knowledge of its nutritional qualities and their impact on hen performance prevents optimization of its use. The present study investigated the digestibility of amino acids ( AA: ) in triticale grain in laying hens, and additionally examined relationships between AA digestibility and chemical and physical characteristics of the grain. Twenty genotypes of triticale were grown under standardized agronomic and environmental conditions and were characterized according to their physical properties (thousand-seed weight, test weight, falling number, extract viscoelasticity), chemical composition (proximate nutrients, non-starch polysaccharides, AA, minerals, inositol phosphates) and gross energy concentration. Additionally, the in vitro solubility of nitrogen was determined. The animal trial comprised 4 Latin Squares (6 × 6) distributed among 2 subsequent runs. Twelve cecectomized LSL-Classic hens were individually housed in metabolism cages and either fed a basal diet containing 500 g/kg cornstarch or one of 20 triticale diets, each replacing the cornstarch with one triticale genotype, for 8 d. During the last 4 d, feed intake was recorded and excreta were collected quantitatively. Amino acid digestibility of the triticale genotypes was calculated by linear regression. The digestibility of all AA differed significantly between the 20 genotypes, including Lys (digestibility range 68 to 80%), Met (77 to 86%), Thr (68 to 78%) and Trp (74 to 83%). However, AA digestibility only correlated with characteristics of the grain in few cases, without a consistent pattern among AA. Equations to predict AA digestibility based on the grain's physical and chemical characteristics were calculated by multiple linear regression. The explanatory power (adjusted R2;) of these prediction equations was below 0.7 for most AA and thus not sufficiently precise to be suitable for practical application. In conclusion, AA digestibility of triticale grain is high overall in laying hens but varies significantly between crop genotypes. This variation could not be well explained by physical and chemical characteristics of the grain.

Pressor action of centrally perfused angiotensin II in rats with hereditary hypothalamic diabetes insipidus.

A method was devised for the perfusion of the cerebral ventricles in conscious rats. Using this method a basal secretion rate of 15 +/- 3 pg of immunoreactive angiotensin II per min was calculated. This material was suggested to be of extrarenal origin. In comparison to findings in normal Long-Evans rats, pressor responses to intraventricular perfusions of angiotensin II were reduced in rats heterozygous for hypothalamic diabetes insipidus and virtually absent in rats homozygous for the hypothalamic deficiency whether they were treated with vasopressin or not. The pressor response to intraventricular angiotensin II is suggested to be related to the release of vasopressin.

Normotensive and spontaneously-hypertensive rats show differences in sensitivity to arginine-vasopressin as a modulator of noradrenaline release from brainstem slices.

The effect of arginine-vasopressin (AVP) on potassium ion-evoked release of [3H]noradrenaline (NA) from rat brainstem slices was investigated. In normotensive Sprague-Dawley (SD) and Wistar Kyoto (WKY) rats, AVP inhibited release of [3H]NA in a dose-dependent fashion, the magnitude and time course of inhibition at 10(-12)M AVP being similar to that observed using the alpha 2-adrenoceptor agonist, clonidine at 10(-7) M. However, it is unlikely that AVP functions through alpha-receptors since its effect is not blocked by phentolamine (10(-6) M). When brainstems from spontaneously-hypertensive (SH) rats (stroke-prone strain) were used, AVP was found to be without effect on [3H]NA-release at concentrations up to 10(-8) M. The potassium ion-evoked release appeared to have two components. The early phase of release was tetrodoxotin (TTX)-sensitive and was probably due to action potential conduction within the slices. The later phase, which was unaffected by tetrodotoxin, was probably a result of potassium-induced depolarization of nerve endings. AVP appeared to affect principally the early, TTX-sensitive release. In the presence of TTX (0.3 microM) the inhibitory effects of AVP were no longer seen. Thus, AVP may affect generation or conduction of action potentials within this tissue. There is published evidence that central vasopressinergic neurons may play a role in controlling the baroreceptor reflex arc. The present study provides a possible biochemical basis for this effect and is consistent with a neuromodulatory role of AVP in the CNS.

Angiotensin-induced drinking in rats with hereditary hypothalamic diabetes insipidus.

Previous reports indicate that the pressor response seen following intracranially applied angiotensin II (AII) in normal rats is absent in hereditary hypothalamic diabetes insipidus (DI) animals. The dipsogenic effect of intracranially injected AII was therefore examined in DI rats and in control rats. Our results show that the drinking response to AII was present in DI animals and not different from control rats. Substitution of DI animals with vasopressin had no effect on the drinking response to AII. We conclude that it is possible to dissociate brain mechanisms mediating the centrally induced blood pressure response and the drinking response.

Decreased vasopressin content in brain stem of rats with spontaneous hypertension.

The content of arginine vasopressin in the brain stem and neurohypophysis of adult spontaneously hypertensive rats was measured by radioimmunoassay and the values were compared to those measured in normotensive Wistar-Kyoto rats. In the brain stem of hypertensive rats, AVP content was decreased by 77% while neurohypophyseal AVP content was increased by 26%. The results demonstrate that brain stem AVP levels and neurohypophyseal AVP levels may change differentially and they are consistent with the possibility that brain stem AVP may be involved in altering cardiovascular reflex activity.

Pathogenesis of malignant hypertension: experimental evidence from the renal hypertensive rat.

In rats with unilateral renal artery stenosis, the malignant phase of hypertension is characterized by: systolic blood pressure above 180-190 mm Hg; sodium and water loss; polyuria and polydipsia; markedly activated renin-angiotensin-aldosterone system; impairment of renal function and malignant nephrosclerosis in the contralateral kidney; some rats exhibit signs of cerebral hemorrhage, heart failure, acute renal failure, and some rats die. After such a phase of malignant hypertension, a period of remission may occur, which is followed by another malignant phase, etc. When malignant hypertensive rats are offered, in addition to water, saline as drinking fluid, they compulsively drink the saline, BP falls transiently, and all signs of malignant hypertension nearly or completely disappear. These observations indicate that, at a critically high BP level, it is salt and water loss which, by activating the renin-angiotensin system, trigger the vicious circle of malignant renal hypertension in rats.