Fibre-based feed supplements - fermentation and moisture characteristics, edibility and particle size influence cage-reared laying hens' preference based on feed consumption and time at the feeder.

1. Past studies have shown that fibre-based feed supplements, such as silage, are well accepted by laying hens and can reduce feather pecking and cannibalism. What is uncertain is whether fermentation and moisture characteristics, edibility or particle size influence the hen's choice for a fibre-based feed supplement or if there are more preferred materials.2. The study included three experiments on fermentation and moisture characteristics (Experiment 1), edibility (consumable; Experiment 2) and particle size (Experiment 3) on laying hen preference for different supplements.3. Experiments were conducted in conventional cages, where two cages formed one replication (six replicates per treatment) and each feeding area was subdivided into a trough (basal diet) and a supplement insert (supplements). Since the hens had a free choice between the basal diet and the supplements, feed consumption and the percentage of time hens spent at the supplement insert indicated the strength of preference. The basal diet dry matter (DM) consumption was assessed for all experiments and the supplement and total DM consumption was documented for Experiments 1 and 3. In addition, the percentage of the observed time hens spent at the trough or supplement insert was observed for Experiments 2 and 3.4. There was an increase in the supplement DM consumption for non-fermented and moist supplements (P < 0.05) and, in some cases, a smaller particle size (P < 0.05). Furthermore, hens spent more time with edible (P < 0.05) and small-sized (P < 0.05) supplements. It was concluded that a preferred material in addition to the basal diet could increase the time hens spend at the feeder by up to 1 hour per photoperiod.

Development of a Coccidiosis Disease Challenge Model Using a Commercially Available Live Oocyst Vaccine.

With growing cross-disciplinary collaboration among researchers, it is increasingly important to record detailed methodology to prevent the repetition of preliminary experiments. The purpose of this paper is to explain the development of a coccidiosis challenge model for the investigation of dietary interventions to coccidiosis in broiler chickens. The objectives are to select a dose of mixed species coccidial vaccine and evaluate the suitability (ability to produce a consistent, marked change) of selected response variables important to nutritional studies at different times postinfection (PI). Coccivac-B and Coccivac-B52 (Merck Animal Health) were evaluated as the source of coccidia in three trials. Trials 1 and 2 were randomized complete block designs with four doses (0, 10, 20, or 30 times (×) label dose) of Coccivac-B administered to 12 replicate cages of six birds by repeater pipette (Trial 1) or gavaging needle (Trial 2). Trial 3 used a completely randomized design with 0× or 30× label dose of Coccivac-B52 administered by gavaging needle to six replicate cages of six birds. Birds were gavaged at 15 days of age, and response criteria were evaluated 7 days PI in all trials and again at 10 days PI in Trials 1 and 2. All means are reported in order of increasing coccidia dose with significance accepted at P ≤ 0.05. Broiler performance was not affected by coccidia in Trials 1 or 3 but grew poorer with increasing dose from 0 to 7 days PI in Trial 2 (body weight gain, 465, 421, 388, 365 g; feed to gain, 1.37, 1.47, 1.52, 1.58). As coccidia dose increased, nitrogen corrected apparent metabolizable energy decreased (Trial 1, 3387, 3318, 3267, 3170 kcal kg-1; Trial 2, 3358, 2535, 2422, 2309 kcal kg-1; Trial 3, not measured), while relative weight, length, and content for intestinal sections increased (Trials 1through 3). Gross lesion (duodenum, jejunum/ileum, ceca) and oocyst count scores (jejunum/ileum, ceca) increased with dose; however, gross scoring often suggested infection in unchallenged birds, a finding unsupported by oocyst count scores. At 7 days PI there was no correlation between midgut gross lesion score and midgut oocyst count score (r = 0.06, P = 0.705), but cecal scores were weakly correlated (r = 0.55, P < 0.001). Administering coccidia via repeater pipette (Trial 1) resulted in respiratory distress in some birds, while use of the gavaging needle (Trials 2 and 3) successfully induced intestinal damage in chickens without resulting in coccidia related mortality. Thirty times the label dose at 7 days PI resulted in the greatest number of response variables that produced a consistent, marked change. Therefore, consideration should be given to these conditions when designing future coccidiosis challenge models using vaccines as a source of coccidia.

Artículo regular­Desarrollo de un modelo de desafío para coccidiosis utilizando una vacuna de ooquistes vivos disponible comercialmente. Con la creciente colaboración interdisciplinaria entre investigadores, es cada vez más importante registrar la metodología detallada para evitar la repetición de experimentos preliminares. El propósito de este artículo es explicar el desarrollo de un modelo de desafío de coccidiosis para la investigación de intervenciones dietéticas para coccidiosis en pollos de engorde. Los objetivos son seleccionar una dosis de vacuna coccidial de especies mixtas y evaluar la idoneidad (capacidad de producir un cambio marcado y consistente) de las variables de respuesta seleccionadas que son importantes para los estudios nutricionales en diferentes momentos posteriores a la infección (PI). Las vacunas Coccivac-B o Coccivac B-52 (Merck Animal Health) se evaluaron como fuente de coccidias en tres ensayos. Los ensayos 1 y 2 fueron diseños de bloques completamente aleatorios con cuatro dosis (0, 10, 20 o 30 veces (×) la dosis indicada en la etiqueta) de Coccivac-B administradas a 12 jaulas repetidas de seis aves mediante una pipeta repetidora (ensayo 1) o por sonda oral. (Prueba 2). El ensayo 3 utilizó un diseño completamente aleatorio con una dosis de etiqueta de 0 × o 30 × de Coccivac-B52 administrada con una sonda oral en seis jaulas repetidas de seis aves. Las aves fueron inoculadas por sonda a los 15 días de edad y los criterios de respuesta se evaluaron a los 7 días postinoculación en todos los ensayos y nuevamente a los 10 días postinoculación en los ensayos 1 y 2. Todos los promedios se reportan en orden de dosis crecientes de coccidias con significancia aceptada en P ≤ 0.05. El rendimiento de los pollos de engorde no se vio afectado por las coccidias en los Ensayos 1 o 3, pero empeoró al aumentar la dosis de los cero a 7 días después de la inoculación en el Ensayo 2 (aumento de peso corporal, 465, 421, 388, 365 g; alimento para ganar, 1.37, 1.47, 1.52, 1.58). A medida que aumentaba la dosis de coccidia, la energía metabolizable de nitrógeno aparente y corregida disminuyó (Prueba 1, 3387, 3318, 3267, 3170 kcal kg-1; Prueba 2, 3358, 2535, 2422, 2309 kcal kg-1; Prueba 3, no medida), mientras que el peso relativo, la longitud y el contenido de las secciones intestinales aumentaron (ensayos 1 a 3). La lesión macroscópica (duodeno, yeyuno/íleon, ciego) y las puntuaciones del recuento de oocistos (yeyuno/íleon, ciego) aumentaron con la dosis; sin embargo, la puntuación bruta a menudo sugirió infección en aves no desafiadas, un hallazgo que no está respaldado por las puntuaciones del recuento de ooquistes. A los 7 días después de la infección no hubo correlación entre la puntuación de la lesión macroscópica del intestino medio y la puntuación del recuento de oocistos del intestino medio (r= 0,06, P= 0,705), pero las puntuaciones cecales se correlacionaron débilmente (r = 0.55, P <0.001). La administración de coccidias a través de una pipeta repetidora (Ensayo 1) provocó dificultad respiratoria en algunas aves, mientras que el uso de la sonda oral (Ensayos 2 y 3) indujo con éxito el daño intestinal en los pollos sin dar como resultado mortalidad relacionada con los coccidias. Treinta veces la dosis de la etiqueta a los 7 días después de la infección resultó en el mayor número de variables de respuesta que produjeron un cambio marcado y consistente. Por lo tanto, deben tenerse en cuenta estas condiciones al diseñar futuros modelos de exposición a la coccidiosis que utilicen vacunas como fuente de coccidias.

Contrasting the effects of phytase and pure myo-inositol on the performance, digestibility, blood and egg yolk inositol levels and digestion physiology of laying hens.

ABSRACT1. An experiment was designed to compare the effects of supplementing laying hen diets with phytase and myo-inositol (inositol).2. Five diets were formulated: high balanced protein (HBP - 840 mg of Dlys/hen/day), HBP with inositol (HBP+I - 0.16%), reduced balance protein (RBP - 672 mg of Dlys/hen/day), RBP with inositol (RBP+I - 0.16%) and RBP with phytase (RBP+P - 3000 FTU/kg).3. Laying hen production, inositol concentrations, digestive tract morphology, amino acid digestibility and intestinal inositol transporters transcript abundance were evaluated. Data were analysed with a one-way ANOVA in SAS 9.4. Contrasts were used to assess the effect of protein, inositol, phytase and phytase vs. inositol. Differences were accepted when P ≤ 0.05.4. No effect on hen-day egg production or feed efficiency was found. However, feed intake and the incidence of abnormally shaped eggs were 0.77 g/h/d and 0.17% higher, respectively, in inositol treatments. Inositol decreased egg specific gravity from 1.088 to 1.0865.5. Inositol concentration in egg yolk was similar among HBP+I, RBP+I and RBP+P, and higher than for the HBP and RBP diet groups. Both gizzard and ileal digesta were enriched in inositol in all supplemented treatments, and phytase supplementation decreased the level of IP5 and IP6 in the gizzard and ileum. Generally, neither phytase or inositol affected amino acid digestibility.6. Inositol increased transcript abundance of alkaline phosphatase in the ileum, while phytase upregulated duodenal alkaline phosphatase and SMIT1, jejunal SMIT2 and reduced ileal HMIT and SMIT1 abundance.7. In conclusion, no effect of phytase or inositol was found for laying hen production performance or amino acid digestibility, but egg quality was reduced by inositol supplementation. Inositol concentration in egg yolk was similar among supplemented treatments.

Evaluation of inclusion level of wheat distillers dried grains with solubles with and without protease or ß-mannanase on performance and water intake of turkey hens.

It is becoming a common practice to use higher levels of wheat distillers dried grains with solubles (wDDGS) in poultry diets. The objective of this study was to determine the effects of level of inclusion of wDDGS with or without enzyme (E-, i.e., wDDGSE-) supplementation on performance and water consumption of turkey hens (0 to 72 d). Two diets (0 or 30% wDDGS) were formulated to meet the nutrient requirements of Hybrid Converter turkeys. Diets (0 or 30% wDDGS; starter, grower, and finisher) were then blended to obtain a different level of inclusion (15%) of wDDGS. The 30% wDDGS diet was divided into 3 fractions and 2 fractions supplemented with either protease (P+, i.e., wDDGSP+; 0.126 g/kg) or ß-mannanase (M+, i.e., wDDGSM+; 0.05 g/kg). All 5 diets were fed ad libitum as mash. The 700 0-d turkey hens were randomly allocated into groups of 35 birds per replicate with 4 replicate floor pens per treatment, in a completely randomized design. Water consumption per pen was recorded beginning at 7 d. There was no effect of dietary treatment on feed intake. BW of turkey hens (52 d; grower) was significantly higher for 30% wDDGSP+ as compared to 0% wDDGSE- or 15% wDDGSE- diets; but was not different from 30% wDDGSE- or 30% wDDGSM+ diets. FCR (P < 0.01; 28 to 52 d), and total FCR (P < 0.05; 0 to 72 d) was significantly improved for birds fed 15 or 30% wDDGS regardless of enzyme treatment compared to 0% wDDGSE-. Water intake (WI, in mL per bird per day) tended to be higher (P = 0.08) between 7 and 28 d for 30% wDDGSP+ diets compared to other treatments. Similarly, WI of birds fed 30% wDDGSP+ was higher (P < 0.05; 28 to 52 and 52 to 72 d) and total WI (P = 0.07; 7 to 72 d) tended to be higher than other treatments. This study is the first to report the impact of wDDGS on WI. As high as 30% wDDGS can be substituted in turkey hen diets. No effect of P+ or M+ at the inclusion level tested was found on performance.

The effects of extrusion of wheat distillers dried grains with solubles with or without an enzyme cocktail on performance of turkey hen poults.

Two experiments were conducted to determine if extrusion (EX) or enzymes (E) could overcome the restrictions (e.g., high fiber) of feeding wheat distillers dried grain with solubles (wDDGS) and improve its nutritional value for feeding turkeys. Two starter diets with either 0 or 30% wDDGS were formulated to meet or exceed the nutrient requirements of the Hybrid Converter female turkeys. The 30% wDDGS diet was substituted with either non-extruded (EX-) or extruded (EX+) wDDGS to produce three basal diets [0% wDDGS (EX-) or 30% wDDGS (EX-/EX+)]. Diets were blended to obtain 15% wDDGS. In the respective treatments, only wDDGS was extruded (temperature; 118°C, retention; 15 sec, total moisture; 25% and pressure 33 bar). The respective experimental diets were supplemented with/without an enzyme cocktail (E; 0.5 g/kg). Test diets were fed from 7-21 d in a completely randomized design. In Experiment 1, a total of 210 turkey hen poults were fed diets containing 0, 15, or 30% wDDGS (EX-) with or without enzyme (E+/E-). Body weight (BW) and feed intake (FI) were significantly higher for 0% wDDGSE-. Nitrogen retention (NR) and apparent metabolizable energy (AME) for the 30% wDDGSE- was significantly higher than other treatments at 21 d. The results indicated significant main effects of E and an interaction between wDDGS level and E. In Experiment 2, 280 turkey hen poults were fed 8 diets [15/30% wDDGS (E+/E-), (EX-/EX+)]. The level of wDDGS had a significant effect on BW, FI and gain:feed; 15% inclusion was superior to 30%. There were significant 2- and 3-way interactions for AME and NR at 21 d due to differences in enzyme response with 15 or 30% wDDGS inclusion and/or extrusion of wDDGS. As high as 15% wDDGS can be incorporated in turkey hen diets. There were no beneficial effects of EX or E on poult performance.

Effect of Quantum phytase on nutrient digestibility and bone ash in White Leghorn laying hens fed corn-soybean meal-based diets.

The efficacy of an Escherichia coli 6-phytase supplementation (Quantum) on nutrient digestibility-retention and bone ash in laying hens fed corn-soybean meal (CSM) diets was investigated. White Leghorn hens (Shaver and Bovan strains) were fed CSM diets containing 0.35% (positive control, PC), 0.25% (negative control 1, NC1), or 0.15% (negative control 2, NC2) nonphytate P from 21 to 61 wk of age. Six more diets were manufactured by supplementing the negative control diets with 200, 400, and 600 units per kilogram of exogenous phytase resulting in a total of 9 treatments. Each dietary treatment x strain subclass was replicated twice with 6 hens per replication. Fecal and ileal digesta samples were collected at 42 wk of age to determine apparent nutrient digestibility or retention. Left tibiae were collected at 42 and 61 wk of age to determine bone ash. The coefficients for ileal digestibility and fecal retention for protein were higher (P < 0.05) for the unsupplemented negative control treatments compared with the PC. A linear reduction in phytate digestibility and ileal protein digestibility was reported with increasing levels of phytase to the NC1 diet. Phytase addition to the NC1 treatment resulted in a linear decrease in the digestibility of amino acids except for methionine and proline. Significantly higher phytate digestibility was demonstrated with the NC2 treatment containing 400 units per kilogram of phytase compared with the PC. Tibial bone ash percentage was higher (P < 0.05) in 61-wk-old hens fed 200 or 400 units per kilogram of phytase-supplemented NC2 diets. Significantly higher diet AME and fecal protein retention were demonstrated for Shaver hens in comparison to the Bovan hens. Overall, the Quantum phytase was not efficacious at improving nutrient digestibility-retention in laying hens fed CSM diets deficient in nonphytate P.

The efficacy of quantum phytase in a forty-week production trial using white leghorn laying hens fed corn-soybean meal-based diets.

Microbial phytase is a prominent feed enzyme used in animal feeds, but there is relatively little information on its use in laying hen diets. In this experiment, an Escherichia coli 6-phytase (Quantum) was evaluated for its efficacy in a 40-wk laying hen production trial. A total of 1,080 White Leghorn hens (540 each of Shaver and Bovan strains) were fed mash corn-soybean meal-based diets containing 0.35% (positive control, PC), 0.25% (negative control, NC1), or 0.15% (NC2) nonphytate phosphorus (NPP). Six more diets were manufactured by supplementing the negative control diets with 200, 400, and 600 U/kg of exogenous phytase, resulting in a total of 9 treatments. Each dietary treatment x strain subclass was replicated 4 times with 5 adjoining cages per replicate (3 hens per cage) in a randomized complete block design. Production performance was measured from 21 to 61 wk of age. Only minor differences in production characteristics were found between the PC and NC1 treatments regardless of phytase addition, indicating that 0.25% NPP resulted in P intake that was at or above the hen's requirement. In contrast, the hens fed 0.15% NPP diet without phytase supplementation had significantly (P < 0.05) reduced total hen housed egg production and body weight at 61 wk of age in comparison to the PC treatment, whereas the incidence of soft-shelled, cracked, and broken eggs was increased significantly (P < 0.05) in hens fed the NC2 diet. Addition of phytase to the NC2 diet improved these production characteristics to levels equal or better than the PC diet. The results indicated that Quantum phytase was efficacious in corn-soybean meal-based diets fed to White Leghorn laying hens and can be used to reduce diet supplementation with inorganic phosphorus.

Feeding high levels of lupine seeds to broiler chickens: plasma micronutrient status in the context of digesta viscosity and morphometric and ultrastructural changes in the gastrointestinal tract.

This work examines the effects of lupine-based diets on the status of the host's riboflavin and zinc. Test diets contained 35% soybean meal (control) or raw (40%) or dehulled (35%) lupine seed meal and were isocaloric (13.4 MJ of AME/kg) and isonitrogenous (23% crude protein). Each diet was offered ad libitum to a group of 16 male commercial broiler chicks for 21 d, starting at 1 d of age. Broilers fed lupine diets had lower feed intakes and growth rates. All sections of the intestinal tract were significantly enlarged (P < 0.01) in all groups fed lupine-based diets in comparison with broilers fed the soybean meal diet, but there were no differences in the morphologies of the mucosa, submucosa, muscularis, or serosa. Increased size of the duodenum, jejunum, and ileum was predominantly attributed to the increase in length rather than intestinal tissue mass. Hence, the enlargement of the intestinal tract was consistent with physiological hyperplasia and not pathological remodeling and hypertrophy. Overall, broilers fed lupine-based diets had more viscous digesta than those fed the soybean meal diet, but the differences were statistically not significant. Blood plasma Zn concentration did not differ between broilers fed lupine-based diets and those fed soybean-meal-based diets, and all broilers fed lupine-based diets had significantly higher (P < 0.001) riboflavin concentrations. In this context, it is apparent that the bioavailability of these micronutrients from lupine diets is not compromised. Intestinal tissue hyperplasia may be interpreted as physiological adaptation to increase absorptive capacity and thus maximize absorption of essential nutrients in the face of antinutritional factors in the diet.

A short introduction to the new principle of binding ration calcium with sodium zeolite.

This paper summarise the development of the new principle of preventing parturient hypocalcaemia by reducing the bioavailability of ration calcium with calcium binders, based on the idea that a negative calcium balance would stimulate natural defence mechanisms against threatening hypocalcaemia. Synthetic sodium zeolite was selected as a first choice among the many calcium binders available commercially, such as polyphosphates, citrate, EDTA and it derivatives. Testing was done on non-pregnant rumen fistulated cows in the first place, followed by cows in late lactation. Encouraged by the tendencies seen in these animals, the final proof of concept was done on pregnant dry cows fed a supplement of synthetic sodium zeolite A from 4 weeks before expected calving until calving. By analysis of blood calcium levels, this supplementation was shown to have a stabilizing effect during the critical period shortly after calving.

Effect of dietary calcium, 25-hydroxycholecalciferol, or bird strain on small intestinal phytase activity in broiler chickens.

Broilers chickens have limited ability to utilize phytate phosphorus (PP), and the influence of nutrients on the activity and efficacy of intestinal phytase is unclear. Therefore in a 2 x 2 x 2 factorial experiment, male chicks were fed 0 or 0.21 mg/kg 25-hydroxycholecalciferol (25-OH D3), 4 or 9 g/kg Ca supplied from CaCO3 or Ca malate from 14 to 24 d of age (six pens/treatment, eight birds/pen). Source of Ca had no effect on tibia ash, intestinal phytase activity, or apparent ileal PP hydrolysis. Phytase activity (Vmax) within brush border vesicles prepared from small intestinal mucosa was greater in chicks fed 4 vs. 9 g/kg Ca (P < or = 0.05). Similarly, birds fed 4 vs. 9 g/kg Ca were able to hydrolyze an additional 24.36% PP (P < 0.01). Intestinal phytase activity and apparent ileal PP hydrolysis were not affected by 25-OH D3 supplementation, but tibia ash was improved by 2.7% (P < 0.01). A subsequent experiment validated the effect of dietary Ca (4 and 9 g/kg) and elucidated differences between strains (Ross 308 and Hubbard x Peterson) from 8 to 22 d of age (six pens/treatment, eight birds/pen). The strains responded similarly to dietary Ca in terms of intestinal phytase and apparent ileal PP hydrolysis. Intestinal phytase activity was 9% greater in birds fed 4 vs. 9 g/kg Ca (P < 0.05) and apparent ileal PP hydrolysis was 11.9% greater (P < 0.02). In conclusion, a typical dietary Ca concentration (9 g/kg) reduced intestinal phytase activity and apparent ileal PP hydrolysis.

The effects of toasting canola meal on body weight, feed conversion efficiency, and mortality in broiler chickens.

It is hypothesized that the moisture incorporated into canola meal (CM) during desolventization, as sparge steam, promotes toasting. Elimination of toasting of CM would result in higher digestible amino acid content, but it is not known if it is required to reduce anti-nutritional factors. Therefore, the objectives of this study were to determine if suspending the use of sparge steam would prevent toasting and to study the effects of toasting on broiler chicken performance. Conventional toasted CM (TCM) and a hexane laden, nontoasted CM (NTCM) were collected from a commercial crushing plant. NTCM was desolventized in a research desolventizer/toaster without the use of sparge steam, producing a yellow-colored meal. The meals were fed to broiler chickens from 0 to 39 d and replaced 0, 20, 40, 60, 80, and 100% of the soybean meal (SBM) in wheat-based diets. Elimination of toasting increased broiler weight from 0.606 and 2.148 to 0.618 and 2.181 kg at 19 and 39 d of age, respectively. The feed ratio (0 to 19 d) increased from 0.637 to 0.642, but toasting did not affect mortality. Total mortality and chronic heart failure between 19 and 39 d increased with level of CM addition from 5.2 to 13.9% and 1.9 to 9.6%, respectively. Chronic heart failure in males, but not females, was increased from 3.3 to 17.4% with increasing CM level. In conclusion, desolventization without sparge steam produced a nontoasted meal and improved broiler growth and feed efficiency in comparison to TCM. Therefore, NTCM could be fed to broiler chickens.

Adverse effects of dietary lupine in broiler chickens.

This study describes the adverse effects of dietary lupines in broiler chickens for which lupine seeds (Lupinus angustifolius) in raw, dehulled, or autoclaved forms were used as a replacement for soybean meal (SBM) in practical diets. Test diets contained 35% SBM (control) or raw (40%), autoclaved (40%), or dehulled (35%) lupine seed meal. All diets were isocaloric (3,230 kcal/kg AME) and isonitrogenous (23% crude protein). Each diet was offered ad libitum to a group of 16 (four replicates with four birds per replicate) day-old male broiler chicks for 21 d. Chemical analysis of lupine seeds showed no detectable levels of mycotoxins, and total alkaloid contents were below 0.01%. Decreased food intake and growth rate were the main signs observed in all birds fed lupine-based diets. These adverse effects were observed during the first week and persisted throughout the trial. Acute signs of toxicity were observed in four chicks fed the diet containing raw lupine seed during the first week of exposure. Initial clinical signs included leg weakness, lack of coordination, and torticollis. In later stages, during Weeks 2 and 3, some birds fed lupine-based diets showed signs of muscle paralysis and skeletal deformity. Postmortem examination did not show gross pathological changes associated with the dietary treatments. Liver microsomal cytochrome P-50 content was higher (P < 0.05) in birds fed the raw lupine-based diet (mean 0.56 pmol/mg protein) in comparison with controls (mean 0.25 pmol/mg protein), which indicated a systemic effect. Based on the present results, it can be stated that high levels of some varieties of sweet lupines in broiler diets may cause significant adverse effects manifested as 1) decreased feed intake and growth rate in most of the birds, and 2) specific signs of acute and chronic toxicity in some individuals.

Persistent antihypertensive effect of oral nitrite supplied up to one year via the drinking water in spontaneously hypertensive rats.

The hypothesis was studied whether the chronic administration of nitrite lowers the blood pressure of spontaneously hypertensive rats (SHR) and prevents secondary hypertension-induced organ lesions. For this purpose totally 96 SHR received 50 to 75 mmol/l NaNO2 or equimolar amounts of NaHCO3 in their drinking water during 4, 8 or 12 months. At each point of time arterial blood pressure, determined with the tail cuff method, was significantly lower in the NaNO2-group in comparison to the controls indicating that no significant tolerance towards nitrite had developed. There was also a tendency towards reduced cardiac hypertrophy and renal atrophy in the NaNO2-group, however without reaching the level of significance. Drinking water containing 75 mmol NaNO2/l was not well tolerated by young rats in contrast to 50 mmol/l. Possible beneficial effects of high dietary nitrate/nitrite levels are discussed with respect to the low frequency of hypertension observed in vegetarians.

The effects of maternal thiamine nutrition on thiamine status of the offspring in broiler chickens.

The response of broiler chickens to a wide range of dietary supplementation of thiamine to broiler breeder diet was studied in order to understand the effects of maternal thiamine nutrition on the status of thiamine indices in the offspring. Thiamine, and thiamine pyrophosphate (TPP) content, and alpha-ketoglutarate dehydrogenase (KGDH) activity were measured in hearts from 20 day old chicken embryos and from chickens at 1, 7, 14, and 21 days of age and in blood at 21 days of age. Total thiamine content in the heart of day old chicks was higher in comparison to 20 day old embryos. Maternal supplementation of thiamine increased heart thiamine in the offspring (p < 0.001), and increased the activity of KGDH in the hearts of day old chicks (p < 0.001), but not in the embryo. The TPP content in the heart increased in response to both maternal and offspring thiamine supplementation (p < 0.001), however the effect of broiler thiamine supplementation was largely independent from the maternal effect. The effect of maternal thiamine nutrition on the offspring's heart KGDH activity was apparent, but the responses to broiler supplementation were dependent largely on the maternal effect. Blood TPP content was not affected by maternal thiamine supplementation (p = 0.39), but thiamine supplementation in the offspring diets increased blood TPP (p < 0.001). Both maternal and offspring thiamine supplementation increased blood free base thiamine content (both p < 0.001). It is concluded from this study that maternal thiamine nutrition affects thiamine status indices and thiamine metabolism of the offspring.

The study of riboflavin requirement in broiler chickens.

Riboflavin status indices in tissues (brain, liver, heart) and blood plasma, and performance parameters were studied in male and female broiler chickens in response to a wide range of dietary supplementation of riboflavin in order to establish the requirement for riboflavin in fast growing modern broilers. The birds fed riboflavin supplemented diets were increasing their body weight at a higher rate than those fed the unsupplemented diet, but this was apparent only during the first stage of growth (days 1 to 21). Supplementation of 2 mg riboflavin per kg was sufficient to support the maximum growth rate. Feed consumption was not affected by different levels of dietary supplementation of riboflavin. The supplementation of riboflavin in the diet increased (p < 0.001) plasma riboflavin level, but the magnitude of response decreased with age. The main component in the tissues was FAD, followed by FMN and riboflavin. Overall, the dietary riboflavin supplementation had highly significant (p < 0.001) effects on tissue FAD, FMN, and riboflavin status, but the effect of supplementation was clearly pronounced only at days 7 and 14, and thereafter the status of FAD, FMN, and riboflavin in the tissues did not differ between unsupplemented and supplemented birds. Neither FAD, FMN, and riboflavin nor GSSG-RED activity correlate with the level of supplementation. Saturation levels of riboflavin in the blood plasma and tissues, corresponded with dietary riboflavin levels of supplementation at 1 to 2 mg per kg. Based on the performance and biochemical data, the dietary requirement of riboflavin for fast growing broilers should be set at a level of 5 mg/kg. The currently recommended allowance of 3.6 mg riboflavin per kg of ration is not sufficient for modern breeds of broiler chickens.

The role of magnesium in the generation and therapy of benign muscle cramps. Combined in-vivo/in-vitro studies on rat phrenic nerve-diaphragm preparations.

Rats received during 3 weeks a Mg-deficient or a Mg-rich diet; Mg-deficient animals revealed hypomagnesemia, cellular K-depletion and Ca-loading. Phrenic nerve-diaphragm preparations were studied; the physiological Tyrode solution contained low or high-Mg concentrations and 0 or 12 mmol lactate/L. Electric stimulation (indirect via the nerve or direct) produced tetanic contractions and increased force at increasing stimulation frequencies. Significantly lower frequencies were needed to elicit these effects when intra- and extracellular Mg levels were low, in comparison to plentiful Mg supply. Comparing unstimulated and stimulated diaphragmatic tissue electrolyte concentrations revealed tissue losses of Mg, K Ca from stimulated tissues which were less pronounced when Mg supply was optimal. These data support the empiric finding that relief from muscle cramps is promptly offered by Mg supplements.

Therapeutic availability of iron administered orally as the ferrous gluconate together with magnesium-L-aspartate hydrochloride.

Since in vitro experiments had excluded interactions between Fe-gluconate (Fe-gluc) and magnesium-L-aspartate hydrochloride (MAH) in aqueous solutions the present in vivo studies seemed to be justified. Animal studies: Rats were kept on magnesium-(Mg)- and iron-(Fe)- sufficient and deficient diets. The intragastral administration of Fe-gluc significantly increased plasma Fe after 3 h, either given alone, or in combination with MAH (inducing hypermagnesemia). Same results were obtained when fortified diets were offered to Fe/Mg-deficient animals. Human studies: The combination of Fe-gluc (2 x 50 mg Fe per day, per os) plus MAH (2 x 7.5 mmol Mg per day, p.o.) was well tolerated by healthy volunteers. Single dose experiments revealed that Fe-gluc alone and in combination with MAH increased plasma Fe levels during 3 h to the same extent. Two groups of pregnant women with moderately reduced hemoglobin levels either received Fe-gluc (out-patients) or its combination with MAH (at least temporarily hospitalised because of preterm labor). Treatments were well tolerated. Hemoglobin levels did not further decrease, as expected without Fe supplements, during the course of pregnancy, thus indicating the therapeutic availability of the electrolytes in both study groups. Progesterone-induced constipation is frequently observed during pregnancy; hence stool softening reported by 50% of the women receiving Fe-gluc plus MAH (versus 33% in the Fe-gluc group) can be regarded as desirable effect. It is concluded that MAH does not interfere with the enteral absorption of Fe-gluc when both electrolytes are orally administered together. Taking both electrolytes together instead of 2 to 3 h apart from each other, as actually recommended, means a less complicated dosage regimen and probably improves compliance.

Magnesium metabolism: basic aspects and implications of ciclosporine toxicity in rats.

In rapidly growing male Sprague-Dawley rats with an initial body weight of 100 +/- 10 g, we investigated how alimentary magnesium (Mg) supply, Mg metabolism and ciclosporine (Ci)-associated nephrotoxicity are interrelated. Food with 100 ppm Mg (1Mg) or 1,000 ppm Mg (stMg) or 10,000 ppm Mg (rMg), Ci 20 mg/kg body weight daily or olive oil were applied for 3 months (n = 10/group). Mg concentrations in various compartments were measured by atomic absorption spectrophotometry. Creatinine clearance (Jaffe), urinary N-acetyl-beta-D-glucosaminidase (NAG) activity (fluorometrically), urinary sodium excretion (flame photometry) and osmolality were measured. Histomorphological examination was done and renal renin expression was studied by monoclonal antibodies. Ci reduced the Mg concentration of the femur under 1Mg (72.6 +/- 9.7 vs. 112.6 +/- 14.3 mmol/kg dry substance, p < 0.05) and under stMg (150.6 +/- 16.6 vs. 194.1 +/- 10.2 mmol/kg dry substance, p < 0.05), thus indicating Ci-related Mg deficiency. This was due to a significant increase in Mg excretion in Ci treatment compared to dietary controls. Under rMg, there was no difference between Ci-treated and control animals. Ci treatment lowered creatinine clearance in 1Mg (1.42 +/- 0.05 vs. 3.02 +/- 0.58 ml/min) and in stMg (1.04 +/- 0.45 vs. 2.18 +/- 0.51 ml/min), NAG/creatinine and urinary sodium excretion were negatively affected by Ci under 1Mg and stMg. Histomorphology showed macrocalcifications due to Mg deficiency and Ci-specific findings, which were markedly enhanced in 1Mg and stMg. Animals with plentiful Mg supply had no functional alterations due to Ci and no or weakly expressed histomorphological lesions. Renin-positive stained cells were higher in Ci-treated animals. This seems to be functionally relevant under 1Mg and stMg, since it was associated with sodium retention and elevated relative heart weight, indicating hypertension. Alimentary or drug-induced Mg deficiency plays a relevant role in the pathophysiology of chronic Ci nephrotoxicity. Our data suggest that Mg supplementation is helpful to reduce Ci toxicity, even if there is 'normal' alimentary Mg intake.

The study of thiamine requirement in broiler chickens.

Thiamine status in response to a wide range (2 to 32 mg/kg) of dietary supplementation of thiamine was studied in male and female broiler chickens in order to understand the manifestation of marginal thiamine deficiency in chickens. Thiamine content and transketolase (TK) activity, were measured in tissues (brain, heart, liver) and blood samples from chickens at 1, 7, 14, 21, 35, and 42 days of age. There was no correlation between body weights and tissue thiamine or TK activity at 3 or 6 weeks of age. Overall thiamine supplementation did not have significant effects on TK activity in the liver (p = 0.22) or brain (p = 0.07). Males had a higher concentration of thiamine than females (p < 0.001) in livers and brains, but the level in the heart did not differ significantly between sexes. There were substantial contrasts among tissues in changes in the tissue thiamine content in response to dietary thiamine levels. Relative to day 1, unsupplemented birds' heart, brain, and liver thiamine increased by 34, 30, and 31% respectively. In response to supplementation, the heart tissue tended to accumulate thiamine at a considerably higher rate than liver or brain. Blood thiamine concentration tended to decline in unsupplemented birds and those supplemented at 2 and 4 mg/kg levels, whereas in birds supplemented with 8 mg blood thiamine level increased at day 7 and remained relatively constant for the remaining part of the trial. Supplementation at the levels of 16 and 32 mg/kg resulted in blood thiamine increasing throughout the duration of the trial. The presented data indicate that there are organ specific differences in the requirement for thiamine. The heart appears to have a higher requirement than the liver or brain. Since there are potential beneficial effects of thiamine on the heart, this subject deserves more detailed investigation.

The effects of continuously or diurnally fed melatonin on broiler performance and health.

An experiment was designed to investigate the effects of dietary melatonin (MEL) on the performance and health of broiler chicks. Chicks raised under constant lighting were fed practical broiler diets in mash form containing 0, 20, or 40 ppm MEL from 0 to 3 wk. A fourth treatment (0 ppm MEL: 6h/d; 20 ppm MEL: 18 h/d) was used to mimic a diurnal light:dark cycle. Commercial broiler grower diets without MEL supplementation were fed from 3 to 6 wk of age. A small dose-related decrease was observed in weight gain (0 to 14 d) and feed consumption (0 to 21 d) due to continuous feeding of MEL. Continuously fed MEL also improved feed efficiency in the first 3 wk vs the control (0 ppm). Growth and feed consumption from 0 to 21 d were not affected by the diurnally fed diet when compared with the control treatment. No effects were found for mortality. However, the incidence of sudden death syndrome (SDS) was numerically less for the diurnal (3.56%) and 40 ppm (3.76%) diets from 0 to 42 d than for the control treatment 5.56%. Skeletal disease was generally unaffected, with the exception of the incidence of crooked toes at 42 d (0 ppm MEL: 8.67% vs diurnal: 4.89%). Dietary MEL fed either continuously or in a diurnal pattern had a minor effect on the performance and health of broiler chickens.