The influence of ageing on muscarinic receptors, β-adrenoceptors and adenylate cyclase activity in the bovine lung

Muscarinic and -adrenoceptors were identified in airway epithelium, smooth muscle and lung parenchyma from Holstein-Friesian calves and cows and were characterized with [³H]quinuclidinyl benzilate and [³H]dihydroalprenolol, respectively. The muscarinic receptor density in the smooth muscle of cows (B _max=4803±245 fmol/mg protein) was 33% greater (p<0.01) than in calves. Low receptor numbers were detected in the epithelium and parenchyma. In both calves and cows, the density of epithelial -adrenoceptors was twice as high as in smooth muscle and parenchyma. The quantity of -adrenoceptors in the tracheal epithelium (B _max=994±83 fmol/mg protein) and smooth muscle (B _max=492±41 fmol/mg protein) in cows was respectively 37% (p<0.001) and 35% (p<0.01) lower than in calves. Adenylate cyclase (AC) assays indicated that the basal and the (–)-isopropylnoradrenaline- (ISO-) stimulated cAMP production were not significantly different between the calves and cows. After stimulation with NaF, significantly higher cAMP production was found in all tissues from cows. Significant correlations were found between absolute AC responses to NaF and -adrenoceptor density in epithelium (r=–0.75,p<0.001) and smooth muscle (r=–0.63,p<0.01). It seems that, in older animals, the production of cAMP is independent of the number of receptors, indicating the presence of fully active compensatory mechanisms.Abbreviations AC adenylate cyclase - cAMP intra-cellular adenosine monophosphate - G-protein GTP binding protein - GTP guanosine 5-triphosphate - ISO isopropylnoradrenaline - M muscarine - QNB quinuclidinyl benzilate     

Effect of beta‐aminoproprionitrile on eggshell formation

1. Eggshells are bioceramic‐biopolymer composites made by a cell‐mediated deposition of an extracellular matrix which drives the organisation of the inorganic phase. Ultrastructurally, eggshells are composed of shell membranes, mammillary knobs, palisade, and cuticle. Shell membranes are two nets of type X collagen‐containing fibrils. On to these membranes, the mammillary knobs, that is, the crystal nucleation sites, are deposited. Type X collagen is highly cross‐linked and insoluble.

2. In order to evaluate the role of type X collagen cross‐linking on eggshell formation, hens were injected with different doses of β‐aminoproprionitrile, which specifically interferes with cross‐link formation.

3. Changes in egg size and shape were observed. Scanning electron micrographs analysis of these eggs demonstrated marked changes in crystal growth and shell membrane structure and arrangement. A dot‐blot analysis, using a monoclonal antibody against chicken type X collagen, shows a dose‐dependent increase in shell membrane collagen extractability.

4. It is concluded that the formation of β‐aminoproprionitrile‐sensitive cross‐links among the type X collagen molecules of the shell membranes play an essential role in normal eggshell formation.     

Muscarinic receptor subtypes, β-adrenoceptors and cAMP in the tracheal smooth muscle of conventional and double-muscled calves

The total muscarinic (M₁ + M₂ + M₃) and -adrenergic receptors in the tracheal smooth muscle of conventional and double-muscled calves were identified and characterized with the non-specific antagonists [³H]quinuclidinyl benzilate ([³H]QNB) and [³H]dihydroalprenolol ([³H]DHA) respectively.Although the quantity of -adrenoceptors in double-muscled calves was 25% lower (p<0.05) than in conventional calves (B _max=327±89 fmol/mg protein), adenylate cyclase assays indicated that the basal adenylate cyclase activity and the (–)-isopropylnoradrenaline (ISO)- and sodium fluoride (NaF)-stimulated values were not significantly different between these calves. However, the density of muscarinic receptors in double-muscled calves was 40% higher (p<0.01) than in conventional calves (B _max=2955±625 fmol/mg protein). Subtypes of muscarinic receptors were studied with [³H]telenzepine (M₁-receptors), [³H]AF-DX 384 (M₂-receptors) and [³H]4DAMP (M₁ and M₃-receptors). It was found that in both double-muscled and conventional calves about 40% of the receptors were of the M₃-subtype, the remaining 60% being M₂-receptors.From these results, it is suggested that inflammation of the respiratory tract in double-muscled calves may be complicated by an imbalance between the cholinergic bronchoconstrictor and the -adrenergic bronchodilator components of the autonomic nervous system.     

The influence of clenbuterol on prostaglandin F2α-induced dyspnoea in calves

Pulmonary function tests were performed in six healthy calves. Prostaglandin F₂ causes severe narrowing of both upper and lower airways (total lung resistance increased, dynamic compliance decreased). Clenbuterol administered intravenously fifteen minutes prior to prostaglandin F₂ aerosol, and in increasing doses (0, 0.4, 0.8, 1.2 g/kg), on days 1, 2, 4 and 6 of the experiment, effectively but not entirely suppressed these responses.These data indicate that -adrenergic receptors are present in the bovine airways and that the use of clenbuterol (0.8 g/kg) may be effective in treating clinical respiratory disease such as bronchopneumonia in calves.     

Growth,viscosity and beta‐glucanase activity of intestinal fluid in broiler chickens fed on barley‐based diets with or without exogenous beta‐glucanase

1. Three groups of birds were fed for up to 35 days on diets containing 500 g barley (cv. Condor) /kg diet, with or without exogenous β‐glucanase, either a commercial preparation or a recombinant endoglucanase.

2. Birds which received diets containing the exogenous enzymes grew faster for the first 3 weeks but after that there was no apparent difference in rate of growth.

3. β‐Glucanase activities in the crop and small intestine of birds given exogenous enzymes were generally higher than those of birds given only the basal diet.

4. Viscosity of intestinal fluid in birds given only the basal diet decreased with age but there was no corresponding increase in β‐glucanase activity. This discounts bacterial β‐glucanase as a contributory factor in the adaptation to β‐glucanase apparent in older birds.     

Effect of early filleting on the texture of breast muscle of broilers stunned with argon‐induced anoxia

1. Eighty broilers were stunned in batches of 10 with less than 1 % oxygen (air displaced by argon) and their chilled breasts were filleted at 2, 3, 5 or 21 h (argon control) post‐mortem. Twenty broilers were electrically stunned and their breasts filleted at 21 h post‐mortem (electrical control).

2. Meat quality variables studied included pH of the pectoralis major muscle measured at 20 min post‐mortem and at filleting, and texture at 24 h post‐mortem.

3. Argon stunning resulted in a rapid early post‐mortem pH fall, but there was no indication of heat toughening in the breast muscles.

4. Filleting at 2 h post‐mortem, after argon stunning, resulted in 5% of the p. major muscles having greater than 4 kg yield force (slightly tough).

5. Although post‐mortem ageing of argon stunned broilers improved the texture of breast muscle, it is inferred that filleting at 2 h postmortem resulted in breast muscle with acceptable texture.     

Some observations on the use of the morantel sustained‐release bolus in first season‐grazing calves on a Belgian dairy cattle farm

Summary

The efficacy of the morantel sustained‐release bolus (MSRB) in controlling gastrointestinal parasites in first‐season grazing calves was evaluated on a dairy cattle farm in Belgium. The calves grazed a pasture which had been used by bolus‐treated animals in the three previous years. The effect of bolus administration was determined with respect to live weight gain, faecal egg shedding, herbage larval counts, serum pepsinogen levels and ELISA antibody titres.

In spite of an incomplete reduction of faecal egg shedding during the first months of the grazing season, bolus administration resulted in the prevention of parasitic gastro‐enteritis in the calves. A weight gain advantage of 35,2 kg of the bolus‐treated animals over the controls was noted already at two months after turnout. This weight gain advantage was maintained until housing.

The usefulness of serum pepsinogen values and ELISA antibody titres as parameters in prevention experiments is stressed. Both serological parameters gave more information concerning infection level than did the faecal egg output and the herbage larval counts.     

The effect of a β-agonist (clenbuterol) on the heart rate,nitrogen balance and some carcass characteristics of veal calves

Friesian bull calves were used to determine whether rapidly growing calves could be used to study the action of a β-adrenergic agonist (clenbuterol). At seventeen days of age calves were allocated to treatment (4 in initial slaughter group; 8 controls and two groups each of 5 calves given 0.1 or 1.0 mg clenbuterol kg⁻¹ milk replacer DM). Three digestibility/nitrogen balance trials were performed and calves were slaughtered between 150 and 170 kg live weight. Treatment with clenbuterol doubled heart rate for approximately 24 and 72 h in calves treated with 0.1 and 1.0 mg kg⁻¹ clenbuterol, respectively, thereafter heart rate was normal. N retention as a proportion of N intake was increased by treatment with clenbuterol, but differences were only significant in calves weighing 110 kg. At the same mean slaughter weight (160 kg) carcasses from clenbuterol-treated calves were heavier (96.3 and 100.1 kg for 0.1 and 1.0 mg kg⁻¹ clenbuterol-treated calves, respectively) compared with controls (92.2 kg). Results suggest that the proportion of muscle was higher and fat lower in the carcasses of treated compared with untreated calves.     

Effect of calves’ supplementation on performance, nutritional and behavioral characteristics of their dams

The effects of supplementation of beef calves on weight gain, intake and digestibility of pasture, milk production and composition of their dams, and behavior of the pair cow–calf were assessed. Fifty-five beef cows with an initial average body weight of 449?±?8 kg and their respective offspring, with an initial average body weight of 138?+?3 kg and aged between 90 and 150 days, were used. Animals were submitted to an experimental period of 112 days. The experimental treatments consisted of: control?=?mineral mixture only, plan 1?=?high protein and high carbohydrate multiple supplement, plan 2?= high protein and low carbohydrate multiple supplement, plan 3?=?low protein and high carbohydrate multiple supplement, and plan 4?=?low protein and low carbohydrate multiple supplement. About 25 and 12.5 % of the protein requirements were supplied by the high and low protein supplements, respectively, and 15 and 7.5 % of total digestible nutrient requirements by the high and low carbohydrate supplements, respectively. Grazing behavior, performance, milk production, milk composition, intake, and digestibility of the cows were not affected (P?>?0.05) by the supplementation of the calves. The supplemented calves had greater (P?<?0.05) performance (as measured by final body weight, average daily gain, and final body condition score), intake and idle time, and lower (P?<?0.05) grazing time, but supplementation did not affect (P?>?0.05) suckling time and suckling frequency. It can be concluded that supplementation affects the behavior and feed intake of calves. However, it does not affect the suckling time and suckling frequency of calves. Additionally, performance, milk production, nutritional characteristics, and behavior of their dams are not affected.     

Effects of dietary supplementation of (β2‐adrenergic agonist clenbuterol on carcase characteristics and some metabolites in ducks

1. Ducks (622 in total) aged 25 d were given diets supplemented with clenbuterol (CL) at 0 (control), 0.5, 1, 2, 3 and 5 mg/kg for 25 d to investigate the effect of dietary CL on muscle and fat deposition and some metabolites in ducks.

2. The mass of the breast muscles was increased by 10 to 31%, while subcutaneous fat plus skin and abdominal fat pad were reduced by 8 to 23% and 20 to 37%, respectively, in the ducks supplemented 1 to 5 mg CL/kg diet.

3. Increased RNA:DNA ratios in the breast muscle, reduced uric acid and increased free fatty acid concentrations in the serum were observed in clenbuterol‐fed ducks.     

The effect of rotational grazing for periods of one or two weeks on the build‐up of lungworm and gastrointestinal nematode infections in calves

Summary

An experiment was carried out with three groups of grazing calves and one housed control group to study the effect of rotational grazing for periods of 1 and 2 weeks on the build up of lungworm and gastro‐intestinal nematode infections respectively. The experiment demonstrated that rotational grazing for periods of I week on six plots prevented the build‐up of heavy lungworm infections. A buildup of heavier lungworm infections was observed in a group that was rotationally grazed for periods of 2 weeks on three plots and a group which remained on one plot throughout the grazing season; there was no difference between these two groups. In all three situations, there was an adequate development of immunity against D. viviparus, as measured by worm recovery after challenge infection at the end of the experiment in comparison with worm recovery of the similarly challenged control group. Neither rotational grazing scheme protected the calves against gastrointestinal helminthiasis, because tracer calves, which grazed for 4 days only in August or October, acquired infections which would have resulted in severe illness or even death if necropsy had been postponed for a week.     

Effect of β-glucosidase on the meat quality and digestibility in broilers

This study investigated the effects of supplementary β-glucosidase on the carcass composition, meat quality, weight of digestive organ and apparent digestibility in male broilers. Two hundred and forty male, 1-day avine broiler chicks were randomly allocated into four treatment groups and fed with corn-soya bean meal supplemented with 0 (control), 0.6, 1.2 and 1.8 U/g of β-glucosidase respectively. The results showed that there were no significant differences (p > 0.05) among groups in carcass composition (percentages of eviscerated yield, half-eviscerated yield, muscle yield of breast and leg). However, adding 0.6 U/g β-glucosidase to the diet not only altered the meat quality by decreasing the drip loss ratio (p < 0.05) and relative lightness (L*) value (p < 0.01), increasing relative redness (a*) value (p < 0.01), but also significantly decreased the pancreas to body weight ratio (p < 0.05), however, with little effect on liver, proventriculus and gizzard to body weight ratio (p > 0.05). The length and width of duodenum villus were not affected by the addition of β-glucosidase, but the coefficients of total tract apparent digestibility of protein and fat increased by 9.02% (p < 0.05) and 7.40% (p < 0.01) respectively; the parameters of ash were not affected by β-glucosidase addition (p < 0.05). This study provided valuable information for evaluation of the effect of supplementary β-glucosidase on the meat quality and digestibility of broilers.     

Effect of dietary DL‐ethionine on growth and liver metabolism

1. The effects of DL‐ethionine, given in the diet or by injection, on growth, hepatic glycogen and lipids, and the activities of some cytoplasmic NAD‐linked dehydrogenases and NADPH‐producing enzymes were investigated.

2. Growth virtually ceased after feeding ethionine for 2 d.

3. Ethionine induced an accumulation of lipid, decreased the activities of malic enzyme and isocitrate dehydrogenase and increased the activity of lactate dehydrogenase.

4. The concentration of glycogen in the liver of ethionine‐fed chickens increased for 3 d in spite of a decrease in food intake, but then decreased.     

Effect of prolonging the non‐productive period on the performance of moulted broiler breeders

1. The effect of prolonging the non‐productive period by offering cracked grain after food withdrawal on the performance of moulted broiler breeders was investigated.

2. Each increase in the non‐productive period resulted in a gradual decrease in egg production but a gradual increase in the percentages of hatching eggs, fertility and hatchability.

3. There were no significant differences between hens fed on cracked grain for 2 or 3 weeks for any of the performance characteristics, except fertility and hatchability. The best fertility and hatchability were observed in hens fed on cracked grain for 3 weeks.     

Effect of Trilostane and Mitotane on Aldosterone Secretory Reserve in Dogs with Pituitary‐Dependent Hyperadrenocorticism

Background

Maximal aldosterone secretion in healthy dogs occurs 30 minutes postadrenocorticotropin (ACTH; 5 μg/kg IV) stimulation. The effect of trilostane and mitotane on aldosterone at that time is unknown.

Objectives

To assess the effect of trilostane and mitotane in dogs with pituitary‐dependent hyperadrenocorticism on aldosterone secretory reserve. To determine if aldosterone concentration correlates with electrolyte concentrations.

Animals

Serum collected from 79 client‐owned dogs and 33 stored samples.

Methods

Client‐owned dogs had ACTH stimulation tests with cortisol concentrations measured at 0 and 60 minutes and aldosterone concentrations measured at 0, 30, and 60 minutes. Stored samples had aldosterone concentrations measured at 0 and 60 minutes. Ten historical clinically healthy controls were included. All had basal sodium and potassium concentrations measured.

Results

The aldosterone concentrations in the mitotane‐ and trilostane‐treated dogs at 30 and 60 minutes post‐ACTH were significantly lower than in clinically healthy dogs; no significant difference was detected in aldosterone concentration between 30 and 60 minutes in treated dogs. However, a significantly higher percentage of dogs had decreased aldosterone secretory reserve detected at 30 minutes than at 60 minutes. At 30 minutes, decreased secretory reserve was detected in 49% and 78% of trilostane‐ and mitotane‐treated dogs, respectively. No correlation was detected between aldosterone and serum electrolyte concentrations.

Conclusions and Clinical Importance

Decreased aldosterone secretory reserve is common in trilostane‐ and mitotane‐treated dogs; it cannot be predicted by measurement of serum electrolyte concentrations. Aldosterone concentration at 30 minutes post‐ACTH stimulation identifies more dogs with decreased aldosterone secretory reserve than conventional testing at 60 minutes.     

Effect of Trilostane on Hormone and Serum Electrolyte Concentrations in Dogs with Pituitary‐Dependent Hyperadrenocorticism

Background

The effects of trilostane on key hormones and electrolytes over 24 hours in dogs with pituitary‐dependent hyperadrenocorticism (PDH) are unknown.

Objectives

To determine the plasma concentration of cortisol, endogenous adrenocorticotropic hormone (ACTH), aldosterone, sodium, potassium, and ionized calcium concentrations, and plasma renin activity over a 24‐hour period after administration of trilostane to dogs with well‐controlled PDH.

Animals

Nine dogs (mean age 9.3 ± 0.67 years, mean weight 31.9 ± 6.4 kg) with confirmed PDH.

Methods

Prospective study. Thirty days after the first administration of trilostane, blood samples were taken at −30, 0 (baseline), 15, 30, 60, and 90 minutes, and 2, 3, 4, 6, 8, 12, 16, 20, and 24 hours after administration of trilostane and plasma concentration of cortisol, endogenous ACTH, aldosterone, sodium, potassium, ionized calcium, and renin activity were determined.

Results

Cortisol concentrations decreased significantly (P < .001) 2–4 hours after trilostane administration. From baseline, there was a significant (P < .001) increase in endogenous ACTH concentrations between hours 3–12, a significant increase (P < .001) in aldosterone concentration between hours 16–20, and a significant (P < .001) increase in renin activity between hours 6–20. Potassium concentration decreased significantly (P < .05) between hours 0.5–2.

Conclusion and Clinical Importance

Treatment with trilostane did not cause clinically relevant alterations in plasma aldosterone and potassium concentration. Results suggest that in dogs with PDH, the optimal time point for an ACTH‐stimulation test to be performed is 2–4 hours after trilostane dosing. Future studies are necessary to establish interpretation criteria for a 2‐ to 4‐hour postpill ACTH‐stimulation test.     

Effect of light,vitamin d and dietary phosphorus on egg‐shell quality late in the pullet laying year

1. Eighteen diets supplying all combinations of three phosphorus contents (3.1, 4.0 and 4.8 g non‐phytate P/kg) and six vitamin D supplements (37.5 or 150 μg cholecalciferol/kg; or 16 or 24 μg 25‐hydroxy‐cholecalciferol/kg; or 37.5 μg cholecalciferol/kg with either 16 or 24 μg 25‐hydroxycholecalciferol/kg) were fed to 2 880 pullets of two stocks from 64 to 74 weeks of age. The birds were housed in eight light‐proof rooms, four of which had 24‐h light‐dark cycles (16L : 8D) and four had 28‐h cycles (20L : 8D).

2. The dietary treatments had no significant effect upon food intake, egg output, shell thickness, shell deformation or specific gravity of the eggs.

3. The 28‐h cycle reduced mean rate of lay by 4.5%, increased egg weight by 5.8% and increased shell thickness by 9.4%. The proportion of eggs with shell faults revealed on candling was reduced from 4.1 % to 2.8%.

4. It is concluded from this and other sources that decreasing dietary phosphorus or modifying vitamin D supplements may sometimes lead to increases in shell thickness of the order of 1 to 2%, but that these changes are unlikely to result in a measurable reduction in the proportion of cracked eggs late in the laying year.

5. A 28‐h light‐dark cycle results in a longer and more uniform interval between consecutive ovipositions and thus gives reliable increases in shell thickness which are large enough to reduce the proportion of cracked eggs in many practical situations. Whether it is profitable to use an ahemeral cycle will depend upon the relative prices paid for eggs of different sizes.     

Effect of turning upon the sub‐embryonic fluid and albumen of the egg of the Japanese quail

1. The effects of egg turning upon the formation of sub‐embryonic fluid (SEF), and albumen weight and composition are described up to day 8 of incubation.

2. The density of albumen increased, and the density of yolk sac contents decreased, during the first 5 d of incubation. Failure to turn eggs diminished these changes.

3. The rate of formation of SEF and its maximum weight were lower if eggs were not turned, as was the rate of decrease in albumen weight.

4. The concentration of sodium and chloride, as well as osmolality, were higher in SEF than in thin albumen, and were affected by a failure to turn eggs.

5. Static incubation altered the concentrations of yolk nutrients in SEF.

6. It is concluded that the formation of SEF is primarily dependent upon the transfer of sodium and chloride from albumen to SEF so creating an osmotic force for water movement in the same direction. Turning the eggs promotes this process by ensuring an adequate supply of ions in thin albumen adjacent to the blastoderm.     

Effects of the sex‐linked dwarf gene (dw) on skeletal muscle cellularity in broiler chickens

1. The number and size of muscle fibres and the DNA concentration in m. pectoralis superficialis of normal (Dw⁺ / ‐ ) and dwarf (dw/ ‐ ) White Plymouth Rock females were investigated at the ages of 9 and 52 weeks.

2. Live weight and total meat was reduced by 37.4 and 42.8% in dw/ ‐ as compared with Dw⁺/‐ females at the age of 9 weeks. The suppressive effect of the dwarf allele on the weight of m. pectoralis superficialis was 40.6% (67.8 v. 40.3 g in Dw⁺ / ‐ and dw/ ‐ genotypes). This reduction in weight was accompanied by a similar decrease of DNA content (28.8 v. 17.1 mg, i.e. 40.6%), a decrease in number of muscle fibres (829 × 10³ v. 654 × 10³, i.e. 21.2%) and fibre diameter (30.80 v. 28.95 μm, i.e. 6.0%). In mature hens, the differences between genotypes Dw⁺ / — and dw / ‐ were slightly smaller.

3. The different relationships between the muscle growth traits studied are discussed.     

Effect of age of donor on the responsiveness of dispersed and cultured chicken anterior pituitary cells to GnRH‐I

1. The aim of this study was to devise a method to prepare and culture anterior pituitary cells from juvenile and adult chickens in order to investigate mechanisms controlling gonadotrophin‐releasing hormone‐I (GnRH‐I) ‐induced luteinising hormone (LH) release in vitro.

2. The optimum culture medium for maintaining gonadotroph responsiveness to GnRH‐I was bicarbonate‐buffered and phenol red‐free Medium 199 supplemented with 10% foetal calf serum.

3. Cultured pituitary cells from juvenile chickens were more responsive to GnRH‐I than cells from adult cockerels, while no LH was released in response to GnRH‐I from pituitary cells from laying hens.

4. Cultured pituitary cells from adult chickens of both sexes released LH in response to 12‐O‐tetradecanoyl‐13‐phorbol acetate (TPA), an activator of an enzyme involved in intracellular signalling, protein kinase C.

5. It is concluded that freshly‐dispersed and cultured gonadotrophs from adult chickens do not regain their responsiveness to GnRH‐I as well as freshly‐dispersed and cultured gonadotrophs from juvenile chickens. It appears that the stimulus‐secretion coupling pathway between the GnRH‐receptor and the activation of protein kinase C in gonadotrophs from adult chickens is more easily disrupted by dispersion and culture than in gonadotrophs from juvenile chickens.