D. M. Denbow

Response of broilers to graded levels of microbial phytase added to maize–soyabean-meal-based diets containing three levels of non-phytate phosphorus

Male 1-d-old broilers (n 920) were given 0, 200, 400, 600, 800, 1000 and 1200 U microbial phytase/kg diet in combination with 2.0, 2.7 or 3.4 g non-phytate P (nP)/kg or 4.0, 5.1 or 5.8 g total P (tP)/kg in a 21 d trial to assess the effectiveness of phytase in a maize-soyabean-meal diet. In addition to the above twenty-one diets, a positive control P diet supplied 4.5 g nP/kg, 6.9 g tP/kg and 10 g Ca/kg. The basal diet contained 230 g crude protein/kg, 8.8 g Ca/kg, 4.4 g tP/kg and 2.0 g nP/kg. Defluorinated phosphate and limestone were used to supply P and Ca. A Ca:tP ratio of 2:1 was maintained except in the positive control diet which had a ratio of 1.45:1. Phytase additions linearly increased (P < 0.01) body-weight (BW) gain, feed intake, toe ash percentage, and apparent retention (% of intake) or total amount (g/bird) of retained Ca and P, and linearly decreased (P < 0.01) P excretion (g/kg of DM intake) at each level of nP with the magnitude of the response inversely related to the level of nP. Above-normal mortality was only observed in the group receiving 2.0 g nP/kg diet without phytase. Adding nP linearly increased (P < 0.01) BW gain, feed intake, toe ash percentage, Ca retention, total amount (g/bird) of P retained, and P excretion, and linearly decreased (P < 0.01) apparent retention (%) of P. Derived linear and non-linear equations for BW gain and toe ash percentage at the two lower nP levels, 2.0 and 2.7 g/kg, were used to calculate P equivalency values of microbial phytase. The results show that 939 U microbial phytase is equivalent to 1 g P from defluorinated phosphate in broilers fed on maize-soyabean-meal diets. The amount of P released per 100 U phytase decreased as the total amount of phytase increased.

Secretion of active recombinant phytase from soybean cell-suspension cultures

Phytase, an enzyme that degrades the phosphorus storage compound phytate, has the potential to enhance phosphorus availability in animal diets when engineered into soybean (Glycine max) seeds. The phytase gene from Aspergillus niger was inserted into soybean transformation plasmids under control of constitutive and seed-specific promoters, with and without a plant signal sequence. Suspension cultures were used to confirm phytase expression in soybean cells. Phytase mRNA was observed in cultures containing constitutively expressed constructs. Phytase activity was detected in the culture medium from transformants that received constructs containing the plant signal sequence, confirming expectations that the protein would follow the default secretory pathway. Secretion also facilitated characterization of the biochemical properties of recombinant phytase. Soybean-synthesized phytase had a lower molecular mass than did the fungal enzyme. However, deglycosylation of the recombinant and fungal phytase yielded polypeptides of identical molecular mass (49 kD). Temperature and pH optima of the recombinant phytase were indistinguishable from the commercially available fungal phytase. Thermal inactivation studies of the recombinant phytase suggested that the additional protein stability would be required to withstand the elevated temperatures involved in soybean processing.

Feeding, drinking and body temperature of leghorn chicks: Effects of ICV injections of biogenic amines

The effects of dopamine (DA), norepinephrine (NE), epinephrine (E) and 5-hydroxytryptamine (5-HT) on food intake, water consumption and colonic body temperature (Tc) were investigated using Leghorn-type chicks. Food intake was not significantly affected by the intracerebroventricular injections of DA, NE or E. In contrast, 5-HT significantly decreased food intake in both sated and fasted chicks. Tc was significantly decreased by NE and E whereas DA and 5-HT prevented the decline seen in controls. Water consumption was not significantly affected by DA, NE or E; however, 5-HT increased water consumption of sated chicks and decreased water consumption in fasted ones. The feeding response of Leghorn chicks following ICV injection of these biogenic amines differs from that previously observed in the broiler-type chicks. This disparate response is discussed in relation to the different growth rates of the two types of birds.

Neonatally-induced thermotolerance: physiological responses

1. Broiler cockerels exposed to neonatal heat (35-37.8 degrees C for 24 hr) at 5 days of age experienced significantly lower mortality upon exposure to elevated temperatures (35-37.8 degrees C) at 43 days of age than did cockerels not given neonatal heat exposure. 2. No differences were found between neonatally stressed and neonatal control groups in water consumption, core and surface temperature, plasma T3 and T4, protein or glucose concentration when exposed to juvenile heat stress. 3. Heterophil to lymphocyte ratios were significantly lower in the neonatally stressed than in the neonatal control group when exposed to heat at 44 days of age. 4. The results indicate that lower mortality during periods of high environmental temperature in the neonatally stressed chicks may involve homeostatic mechanisms different from those utilized during acclimation to high environmental temperature.

Comparison of brain arginine-vasotocin and corticotrophin-releasing factor for physiological responses in chicks

Arginine-vasotocin (AVT), a non-mammalian homologue of mammalian arginine-vasopressin, is a stress-related peptide in the brain of birds. The aim of the present study was to determine the effects of intracerebroventricular (ICV) injection of AVT on feeding behavior, body temperature, corticosterone release and several behavioral parameters in chicks. These effects were compared with those of corticotrophin-releasing factor (CRF), another stress-related peptide. The ICV injection of AVT inhibited feeding behavior, increased rectal temperature, and increased plasma corticosterone concentrations, but these effects were weaker than those of CRF. AVT induced hypoactivity as evidenced by decreased vocalization and stepping while CRF induced hyperactivity. The present results demonstrate that some functions of brain AVT are similar to those of CRF, although these effects are weaker than those induced by CRF. However, some AVT-induced behaviors were different from CRF, indicating that the physiological roles of AVT in the regulation of stress behavior are different from those of CRF in chicks.

Intracerebroventricular injection of L-arginine induces sedative and hypnotic effects under an acute stress in neonatal chicks.

Summary.l-Arginine participates in many important and diverse biochemical reactions associated with the normal physiology of the organism. In the present study, we investigated the effect of central administration of l-arginine on the stress response and its mechanism in neonatal chicks. Intracerebroventricular (i.c.v.) injection of l-arginine clearly attenuated the stress response in a dose-dependent manner, and induced sleep-like behavior during 10 min. To clarify the mechanism by which l-arginine induces sedative and hypnotic effects in chicks, we investigated the effects of nitric oxide (NO) synthase (NOS) inhibitors on l-arginine-induced sedative and hypnotic effects, and as well as the effects of a NO donor. l-Arginine-induced (1.9 µmol) sedative and hypnotic effects were attenuated by i.c.v. co-injection with a non-selective NOS inhibitor NG-nitro-l-arginine methyl ester HCl (400 nmol). In addition, the effects of l-arginine were slightly attenuated by the inactive isomer of the NOS inhibitor NG-nitro-d-arginine methyl ester HCl (400 nmol). The i.c.v. injection of 3-morpholinosylnomine hydrochloride, a spontaneous NO donor, had little effect on postures. The i.c.v. injection of l-arginine had no effect on NOx concentration at various brain sites. These results suggested that the contribution of NO generation via NOS may be low in the sedative and hypnotic actions of l-arginine. Therefore, l-arginine and/or its metabolites, excluding NO, may be necessary for these actions.

Brain creatine functions to attenuate acute stress responses through GABAnergic system in chicks.

The involvement of brain creatine in the adaptation to acute stress responses was investigated in chicks. In experiment 1, brain creatine content of chicks exposed to social separation stress was significantly increased compared with control chicks. The effects of i.c.v. injection of creatine (2 mug) on vocalizations, spontaneous activity and plasma corticosterone concentration in chicks under social separation stress were investigated in experiment 2. All measurements were attenuated by the i.c.v. injection of creatine compared with the controls under separation stress. Creatine also significantly decreased the active posture, but increased the motionless eye-opened posture, compared with the control. To clarify the relationship between creatine function and GABA receptors, the i.c.v. co-injection of creatine with picrotoxin, a GABA-A receptor antagonist, or CGP54626, a GABA-B receptor antagonist, was investigated in experiments 3 and 4. The effects of creatine on vocalizations and spontaneous activity were attenuated by co-injection of picrotoxin. In this case, active postures decreased by creatine were recovered by co-injection with picrotoxin. However, these effects were not obtained with CGP54626. The results suggest that central creatine functions within the CNS to attenuate the acute stress response by acting through GABA-A receptors in chicks.

Feeding, drinking and temperature responses to intracerebroventricular β-endorphin in the domestic fowl

Four experiments were conducted to evaluate the effect of beta-endorphin (beta-END) on feeding, drinking and colonic temperature in rapidly growing (Rock-Cornish; RC) and slow growing (Single-Comb White Leghorn; SCWL) stocks of chickens. In the first experiment RC cockerels were injected intracerebroventricularly (ICV) with 0, 1.5, 3.0 and 6.0 micrograms of beta-END. In the second experiment RC cockerels were injected ICV with 0.5, 1.0 and 2.0 micrograms of beta-END. Experiments 3 and 4 were conducted identically to Experiment 1 and 2, respectively, except SCWL were used. Administration of beta-END at levels between 1.5 and 6.0 micrograms produced a significant curvilinear increase in feeding in both RC and SCWL chicks. In RC chicks, feeding was significantly elevated at 45 min and from 90 through 240 min postinjection, whereas in SCWL chicks feeding was increased from 90 through 300 min postinjection. Water intake was depressed in RC and SCWL from 60 through 90 min and from 30 through 60 min postinjection, respectively. Significant increases in water occurred at 180 and 300 min postinjection in SCWL. beta-END also induced a significant hyperthermia in RC and SCWL from 30 through 240 min and from 15 through 180 min postinjection, respectively. At low levels of beta-END, i.e., 0, 0.5, 1.0 and 2.0 micrograms, feeding, drinking and body temperature were significantly increased in both stocks. Feeding in RC chicks was stimulated in a linear fashion from 180 through 300 postinjection while feeding in SCWL was stimulated in a curvilinear manner from 180 through 240 min postinjection.(ABSTRACT TRUNCATED AT 250 WORDS)

Centrally and peripherally administered bombesin decreases food intake in turkeys.

The effects of intracerebroventricular (ICV) and intravenous injections of bombesin (BBS) on food intake were investigated in turkeys. Adult turkey hens were injected ICV with 50 to 1000 ng of BBS. In addition, the effect of pretreatment with the BBS antagonist [d-Arg1, D-Phe5, D-Trp7,9, Leu11]-substance P was investigated. To determine if BBS also had a peripheral site of action, 0.5 to 8 micrograms/kg body weight of BBS was injected IV into turkey poults. The ICV and IV injections of BBS decreased food and water intake in a dose-dependent manner. The most efficacious doses when injected ICV for decreasing food and water intake were 1000 and 500 ng, respectively, whereas 8 micrograms/kg was most efficacious in decreasing food intake when administered IV. The satiating effect of ICV-injected BBS could be attenuated with pretreatment with the BBS antagonist. The results of these studies suggest that BBS acts to decrease food and water intake in both the periphery and the central nervous systems of turkeys.

GABAERGIC CONTROL OF FOOD INTAKE IN THE MEAT-TYPE CHICKENS

This study examined the effects of intracerebroventricular injections of gamma-aminobutyric acid (GABA) agonists on short-term food intake in meat-type cockerels. In Experiment 1, birds were injected with various doses of muscimol, a GABA(A) agonist. In Experiment 2, the birds received bicuculline, a GABA(A) antagonist, prior to injection of muscimol. In Experiment 3, the effect of varying doses of baclofen, a GABA(B) agonist, on food intake was determined. The intracerebroventricular injection of muscimol caused a dose-dependent increase in food intake. This effect was significantly attenuated by pretreatment with bicuculline. Food intake was not affected by the intracerebroventricular injection of baclofen. These results suggest that GABA acts within the brain of broilers at a GABA(A), but not GABA(B), receptor to increase voluntary food intake.