Stressor effects of adrenaline on the immature fowl

Chicks were injected with L-adrenaline (500 microgram/kg) thrice weekly from hatching for one, two or three weeks. The growth rate of treated chicks was depressed, particularly in the first week. When four-week-old chicks were given nine injections of adrenaline over a three-week period, growth rate was also depressed but less severely. The greater sensitivity of the younger chicks is thought to result from an immature blood-brain barrier. Relative adrenal mass (mg/kg) was generally increased but adrenal cholesterol depletion occurred only in three-week-old birds treated with adrenaline throughout. Plasma glucose and cholesterole concentrations were within the normal range after three weeks of treatment but there was a significant hypolipacidaemia in the younger birds treated for two or three weeks.     

Effect of daily injections of ACTH on growth and on the adrenal and lymphoid tissues of two strains of immature fowls.

1. The effects of daily injections of ACTH (30 IU/kg) or physiological saline on growth and on adrenal and lymphoid tissues of Rhode Island Red (RIR) and Light Sussex (LS) chickens were compared at 1, 2 and 3 weeks of age. 2. Saline injections retarded skeletal growth in both strains during the 1st week but only affected weight gain in LS birds. 3. Injections of ACTH depressed growth rate, caused adrenal enlargement and deplenished adrenal cholesterol to the same extent in both strains. 4. ACTH caused a greater reduction in bursal size in RIR than in LS birds. Spleen size was reduced by ACTH only in RIR birds. 5. At 2 and 3 weeks the plasma concentrations of corticosterone of RIR birds were greater than those of LS birds. Plasma corticosterone concentrations were within the normal range 24 h after the last injection of ACTH in both strains. 6. Plasma concentrations of growth hormone was unaffected by ACTH treatment in RIR birds, but it was increased in LS birds after 3 weeks of treatment.     

Steroidogenic properties of isolated adrenocortical cells from Japanese quail selected for high serum corticosterone response to immobilization

The effect of selection for high stress response on adrenocortical function was examined by measuring the corticosterone response of adrenocortical cells isolated from random-bred Japanese quail and quail selected for high serum corticosterone response to immobilization (high-stress). Highly enriched adrenocortical cells were incubated with various concentrations of ACTH1-24 (ACTH), 8-bromo-cyclic AMP (8Br-cAMP) and pregnenolone for 2 hr. Corticosterone production was measured by radioimmunoassay. Basal corticosterone production values by cells from random-bred and high-stress birds were not different. In contrast, the average maximal ACTH- and 8Br-cAMP-induced corticosterone production by cells from high-stress quail was 89% greater than that of cells from random-bred quail. However, the average pregnenolone-supported corticosterone production by cells from high-stress birds was 34% less than that of cells from random-bred birds. Thus, the data suggest that although random-bred quail cells had a greater potential capacity for corticosterone production, high-stress quail cells had a greater ability to couple ACTH, ACTH-transmembrane-signaling factors and subsequent second messengers with the available steroidogenic enzyme pool. The magnitude of the differences in function between cells from high-stress and random-bred birds was greater for female cells compared to male cells. In addition to differences in cellular function, there were also differences in adrenal and relative adrenal weights between random-bred and high-stress quail. The average, adrenal and relative (mg/100 g body weight) adrenal weights of high-stress quail were 14-16% greater than those of random-bred quail. It is concluded that the enhanced serum corticosterone response of the high-stress quail line is, in part, due to an increase in relative adrenal weight and an increase in adrenocortical cell responsiveness to ACTH.     

Stressor effects of handling on the immature fowl.

Chicks were handled for 5 min twice daily for five consecutive days per week over a three-week period. The control birds were not handled once they had been allotted to their brooders. The growth rate of the treated birds was significantly decreased. There was no adrenal hypertrophy; the concentration, but not the content, of adrenal cholesterol was significantly increased. Plasma glucose, FFA and cholesterol concentrations were similar in the groups at the end of the experiment. The lipolytic response of the handled birds to a further stimulus of handling was significantly greater than that of unhandled birds experiencing novel handling stimulus. There was also a significant decrease in the concentration of corticosterone in the plasma of the handled birds but not in the unhandled group. There was thus no evidence of habituation.     

Normal sexual maturation and reproductive function in domestic hens following unilateral adrenalectomy

The ovulation-inducing properties of adrenal hormones and the anatomical juxtaposition of the left adrenal gland and the ovary suggest that a functional relationship exists between these two endocrine organs. To test this hypothesis, the effect of unilateral adrenalectomy on sexual maturation and reproductive function was examined in left and right adrenalectomized SCWL pullets. Between 9 and 13 weeks of age, 24 and 12 pullets were either left or right adrenalectomized, respectively; 12 birds were sham operated and 7 birds served as unoperated controls. Blood samples were obtained immediately before and after surgery, 48h after surgery and at 14, 19 and 23 weeks of age. All samples were analyzed for plasma corticosterone by radioimmunoassay. At 20 weeks of age all birds were weighed and the photoschedule was changed from 8L:16D to 16L:8D. During the first 52 days following photostimulation the age at first egg and the distribution of ovipositions within the 24h photoschedule were recorded.Mean plasma corticosterone levels of the unilaterally adrenalectomized hens were not significantly different (P > .05) from those of control birds 48h after surgery or at 14, 19 and 23 weeks of age. Unilateral adrenalectomy had no effect on body weight at 20 weeks, average age at first egg, or percent hen-day production. In addition, 95% of the eggs laid in the first 52 days after photostimulation were restricted to a 10 hour period of the day regardless of the surgical treatment. These observations indicate that the removal of either the left or the right adrenal gland does not affect plasma levels of corticosterone, sexual maturation, or the open period for LH release as evidenced by the times of oviposition and fail to support the hypothesis of a localized functional relationship between the left adrenal gland and the ovary.     

Habituation by the immature fowl in response to repeated injections of corticotrophin.

1. Immature birds treated for 7 d with corticotrophin (30 IU/kg, one injection per day) had significantly poorer growth rates and decreased adrenal cholesterol concentrations. Plasma glucose, corticosterone and cholesterol concentrations and adrenal weight were within the normal range 24 h after the last injection.

2. The responses of birds pretreated as described above to a single injection of corticotrophin (30 IU/kg) were determined.

3. A similar degree of hyperglycaemia had developed in both groups after 2 h but thereafter the responses differed: the concentration of plasma glucose did not increase further in those pretreated with corticotrophin and had begun to decrease at 6 h, whereas that of the birds pretreated with saline increased progressively.

4. The concentrations of plasma corticosterone had increased similarly in the two groups to a peak at 2 h but there was a significantly more rapid decrease in the birds pretreated with corticotrophin.

5. Both groups showed a transient hypercholesteraemia but the increase was significant only in the group that had previously received saline.

6. There were decreases in the concentrations of adrenal cholesterol in both groups. The decrease, in absolute terms, was 2–4 times greater in the group receiving corticotrophin for the first time: percentage changes were similar, however.     

The response of the immature fowl to multiple injections of adrenaline.

Chicks were injected, from one to 21 days of age, with either L-adrenaline (500 mug/kg) or the solvent three times a week. Birds treated with adrenaline showed an imparied growth rate. Relative (mg/kg0.75), but not absolute (mg) adrenal weight was significantly increased. No changes in adrenal cholesterol content or concentration were noted.     

Passive immunization with antiserum to adrenocorticotropin increases weight gain in normal female rats.

Passive immunization against ACTH was used to test the hypothesis that growth in female rats is constrained by physiological concentrations of glucocorticoids. When animals were stressed by 15-min exposure to ether before blood sampling by cardiac puncture, serum concentrations of corticosterone were lower (P less than .05) in immunized rats than in stressed controls. The maximum effect was apparent 2 h after injection of ACTH antiserum, and no effect was apparent 6 h after injection. To examine the effects of ACTH immunization on growth, rats received daily injections of either saline, sheep immunoglobulin G, or ACTH antiserum, 2 h before the afternoon peak in plasma concentrations of corticosterone. After 7 d of treatment, rats treated with ACTH antiserum had gained 37% more body weight than saline-injected controls, and this effect was accompanied by a 59% reduction in peak plasma concentrations of corticosterone. Immunoglobulin G purified from normal sheep serum had no effect on weight gain. It is concluded that growth rate in normal female rats can be stimulated through the suppression of adrenal activity.     

Some responses of the immature fowl to the withdrawal of a stressor

1. Light Sussex chicks were treated with corticotrophin (30 IU/kg) five times a week from hatching to 3 weeks of age. Their responses to the cessation of treatment were determined over the subsequent 2 weeks. 2. Growth rate was impaired by the treatment but had returned to normal after 7 d. 3. Adrenal mass was not affected by treatment but hypertrophy was evident after 14 d of recovery. Relative adrenal weight was greater in the treated group throughout the recovery period. 4. Adrenal cholesterol stores were depleted at the end of treatment, were repleted after 7 d but significantly depleted again at 14 d. 5. Plasma glucose concentration was not affected by treatment but birds were hyperglycaemic 7 and 14 d after treatment had ceased. 6. Plasma free fatty acid (FFA) concentrations were not affected at any time. 7. Plasma corticosterone concentration was not affected by the treatment but 7 d later it was greater in the treated birds. 8. Significant correlations between plasma glucose, FFA and corticosterone were found.     

Development of the chicken as a model for prenatal stress

Exposing a pregnant mammal to stressors causes behavioral and physiological alterations in her offspring ("prenatal stress"); however, elucidation of the underlying mechanism is hindered by an inability to control maternal compounds that may affect the fetus. We designed this experiment to determine if the autonomously developing chicken embryo could be developed as a model for prenatal stress. On d 16 of incubation, eggs were treated with: 1) 60 ng corticosterone (CORT), 2) elevated incubation temperature (40.6 degrees C) for 24 h (HEAT), or 3) no treatment (Control). Chicks from all three treatments hatched at similar weights; however, HEAT chicks weighed less by 100 d of age and remained lighter until the end of the study (P < 0.05). At 8 d post-beak trimming, adrenal gland weight was not different (P > 0.20) among treatments, basal plasma corticosterone concentrations tended (P < 0.06) to be greater for CORT chicks than either the Control or HEAT chicks, and CORT chicks were heavier than HEAT chicks (P < 0.005) but not Control chicks (P > 0.20). At 11-wk, HEAT birds had heavier adrenal glands than did Control birds (P < 0.01). At 16 wk of age, Control cocks performed more (P < 0.01) pecking aggression than either HEAT or CORT cocks, whereas CORT cocks were chased more often and chased another cock less often than either HEAT or Control cocks (P < 0.01). Treatments did not alter the behavior of the hens (P > 0.10). Administration of corticosterone during incubation replicated some, but not all, of the effects seen in prenatal stress in mammals.     

The efficacy of a Giardia lamblia vaccine in kittens.

Twenty kittens were vaccinated with a Giardia lamblia vaccine prepared on a commercial scale on day 0 and boosted on day 21 (group 1); while 10 kittens received only saline (group 2). These kittens were challenged on day 35 with 10(6) Giardia lamblia trophozoites by a surgical intraduodenal injection. Three control kittens were not vaccinated and not challenged (group 3). Following challenge, Giardia vaccinated kittens had significantly fewer days in which abnormal stools were observed and reduced food intake occurred compared to saline injected animals. The rate of weight gain between group 1 and group 2 animals was not different in the prechallenge period (day 0 to day 35), but vaccinated animals had a significantly higher weight gain in the postchallenge period (P < 0.05). On day 56, all vaccinated animals were not passing cysts in their feces, while 40% of saline injected kittens had Giardia cysts in their feces. In vaccinated kittens, cysts were never demonstrated in 45% of the animals, while cysts were detected in 90% of the saline injected kittens. Viability of the cysts in vaccinated kittens was 38% while the cysts viability in saline injected kittens was 99%. On postmortem examination, trophozoites could be detected in 5% of vaccinated kittens and 60% of saline injected kittens. Vaccination produced an elevated Giardia specific serum IgG and IgA response prior to challenge and throughout the postinfection period. The Giardia infection in the saline injected group did not induce an elevated specific serum response. Giardia vaccination of kittens provides protection in kittens from an experimental challenge by reducing or eliminating intestinal trophozoites and fecal cyst excretion.     

A comparison of the oral application and injection routes using the onderstepoort biological products fowl typhoid vaccine, its safety, efficacy and duration of protection in commercial laying hens

This study was undertaken to establish whether the Onderstepoort Biological Products Fowl Typhoid (OBPft) vaccine registered as an injectable vaccine was effective and safe when administered orally to commercial layers. Its efficacy and duration of protection were compared with application by intramuscular injection. Commercial brown layer hens were used as they were found to be highly susceptible to Salmonella gallinarum infections. In the vaccine safety trial birds were euthanased at timed intervals spanning 4 weeks postvaccination. Necropsies were performed and samples were taken and tested. No clinical signs or mortalities could be attributed to the OBPft vaccine nor could active shedding of the vaccine strain be detected. Slight pathological changes were noted with both routes of vaccination; however, these changes were transient, returning to normal within the observation period. The injected groups showed a better serological response with the rapid serum plate agglutination (RSPA) test than the orally vaccinated groups. In the duration of protection trial, birds were challenged at 3-8-week intervals post-vaccination. All unvaccinated birds died. Protection 8 and 16 weeks after vaccination was above 60 %,by 24 weeks after challenge, the vaccine protection was below 30 %. It was found that there was no significant difference (P < 0.05) in the protection offered by either the oral or injected route of vaccination with the OBPft vaccine.     

The immune response of a marine teleost, Pseudopleuronectes americanus, (winter flounder) to the protozoan parasite Glugea stephani

G. stephani is an intracellular cyst-forming microsporidan parasite that is found in the intestine of winter flounder (WF) Pseudopleuronectes americanus. No detectable humoral response was seen in parasitized fish or in fish injected with either spores or spore homogenate from this parasite. Quantification of total immunoglobulin (Ig) levels showed a decrease in Ig levels rather than enhancement, 21 days after intramuscular (IM) injections of spores (3 X 10(6)/ml). When a second injection of spores was administered on day 21 and tested 3 weeks later, a further decrease in total serum Ig's occurred. A decrease in total IgM levels also occurred in WF that were simultaneously injected with G. stephani and the antigens, horse red blood cells (HRBC) or formalin-killed Klebsiella pneumonia (KP). The total Ig levels of fish injected with an antigen plus spores was not as low as those injected with the parasite alone. The Ig levels, as well as antibody titers to HRBC and KP were however, lower when compared to fish injected only with the HRBC or bacteria. Disrupted spore homogenate injected into winter flounder, showed a less marked decrease in Ig levels when compared with whole spores. When a single IM injection of spores was given, followed by two weekly injections of indomethacin (a drug that inhibits prostaglandin activity), no decrease in Ig levels occurred and levels were comparable to control (saline injected) fish.     

Force-feeding procedure and physiological indicators of stress in male mule ducks.

1. The effects of the force-feeding procedure and its different components on various physiological indicators of acute and chronic stress were investigated in male mule ducks before and during a 12-d cramming period. 2. The highest concentration of corticosterone were measured after injection of ACTH agonist, during the pre-experimental period when the ducks were still housed in collective floor pens and at the time of transfer. 3. During the cramming period, corticosterone measured before and after force-feeding did not differ significantly though there was a non-significant trend towards an increase in certain cases. 4. The different components of force-feeding, including manipulation, intubation, force-feeding with a standard or a large amount of food had no significant effect upon corticosterone concentrations. 5. There was no indication from ACTH agonist challenge either of a change in adrenal sensitivity or a change in its responsiveness. 6. The heterocyte-lymphocyte ratio measured before and at the end of the cramming period did not differ significantly. 7. In conclusion, we observed no significant indication that force-feeding is perceived as an acute or chronic stress by male mule ducks, in our experimental conditions. Nevertheless, it remains to be shown that their adrenocorticotropic axis is responsive to acute stressors.     

Effect of exogenous ACTH on serum corticosterone and cortisol concentrations in the Moluccan cockatoo (Cacatua moluccensis)

The effects of exogenous adrenocorticortrophic hormone (ACTH) on the serum corticosterone and cortisol concentrations were determined in 28 mature Moluccan cockatoos (Cacatua moluccensis), a representative of the psittacine species. Birds were randomly assigned to 4 groups (2 ACTH-treated groups and 2 saline-treated controls). Group I (10 cockatoos [5 males and 5 females] ) was given 15 IU of ACTH after blood samples (base line) were taken at 10:00 AM. Blood samples were taken again at 30 minutes and 2.5 hours after ACTH administration. Group II (10 cockatoos) was given similar treatment, but blood samples were taken at 1 and 4 hours after ACTH was administered. Groups III and IV (each of 4 birds) were given saline solution injections as controls. Blood samples were taken at 30 minutes and 2.5 hours after injection (group III) and at 1 and 4 hours after injection (group IV). All serum samples were analyzed for cortisol and corticosterone. Serum corticosterone concentration increased significantly (P less than 0.01) from base-line levels (26 ng/ml) to 108 ng/ml within 30 minutes after ACTH was administered. The high values were maintained for 3 hours and then decreased to 40 ng/ml at the end of 4 hours. Male birds seemed to respond to the ACTH treatment quickly and maintained increased concentration for a shorter period when compared with the responses seen in female birds. Serum cortisol values remained low throughout the experimental period. These results indicate that serum corticosterone was responsive to ACTH administration, but cortisol was not. In addition, there may be a difference in the responses between male and female members of the species.     

Effects of stocking density, flock size and management on the welfare of laying hens in single-tier aviaries

Management practices, stocking rate and flock size may affect laying hen welfare but there have been few replicated studies in commercial non-cage systems that investigate this. This study used a broad range of physical and physiological indicators to assess the welfare of hens in 36 commercial flocks. Six laying period treatments were examined with each treatment replicated 6 times. It was not possible to randomly allocate treatments to houses, so treatment and house were largely confounded. Three stocking rates were compared: 7 birds/m(2) (n = 2450), 9 birds/m(2) (n = 3150) and 12 birds/m(2) in either small (n = 2450) or large (n = 4200) flocks. In addition, at 12 birds/m(2), in both small and large flocks, birds were subjected to either standard (SM) or modified (MM) management. MM flocks had nipple drinkers and no nest-box lights. Bone strength, fracture incidence, heterophil:lymphocyte (H:L) ratio, live weight, organ weights, serum creatine, serum osmolality, muscle pH and faecal corticosterone were measured on samples of birds at the end of the rearing period and at the end of lay. During the laying period, mortality, production and integument condition were recorded at regular intervals. Birds housed at 9 birds/m(2) had higher mortality than birds housed at 12 birds/m(2) by the end of lay, but not higher than birds housed at 7 birds/m(2). Birds housed at 7 and 9 birds/m(2) had lower percent liver weight, and worse plumage condition than most of the 12 bird/m(2) treatments. Modified management tended to improve plumage condition. There were no clear effects of flock size on the welfare indicators recorded. At the end of the rearing period fracture incidence was almost negligible and H:L ratio was within a normal range. By the end of lay fracture incidence was 60% and H:L ratio was high, with no treatment effect for either measure. This, together with information on faecal corticosterone, feather loss and mortality, suggests that the welfare of birds in all treatments was relatively poor by the end of lay.     

Relationship between the milk yield response to short-term bovine somatotropin treatment and the lipolytic response to adrenaline in dairy cows

The aim of this experiment was to determine if the milk yield response of dairy cows to short-term treatment with bovine somatotropin (bST) was correlated with the non-esterified fatty-acid (NEFA) response to an adrenaline challenge. Twenty-six multiparous Holstein cows (58+/-5.4 days postpartum) received daily sub-cutaneous injections of saline for 7 days followed by sub-cutaneous injections of 20mg/day of bST for 14 days. On day 7 of the saline treatment and day 14 of the bST treatment the cows were given an intravenous injection of adrenaline (1.4 microg/kg body weight). Blood samples were taken before and after the adrenaline challenge. The difference in milk yield between the saline and the second week of bST treatment (MYR) varied considerably between animals (from -0.4 to +8.0 kg/day). MYR was positively correlated with the change in the basal concentration of NEFA between the saline and second week of bST treatment, as well as with the change in the area under the profile of NEFA above basal values following the adrenaline challenge. It remains to be established whether the greater lipolytic responses to adrenaline of the cows with the greater MYR reflects the deeper negative energy that these animals also experienced or a fundamental difference in the physiology of their adipose tissue.     

Effect of bipiperidyl mustard and gold thioglucose on the hypothalamus and growth of the hatchling chick and duckling

Following a 48 h fasting period after hatching, male Cobb broiler chicks were injected with either bipiperidyl mustard (intraperitoneally; 0.01 and 0.02 mg/g body weight) or goldthioglucose (intracardially; 0.25 mg/g body weight). The influence of these obesifying agents on growth was compared to control injections of physiological saline. Bi-weekly body weights, and eight week live, carcass, and abdominal fat plus carcass weights were depressed in chicks injected with bipiperidyl mustard, the depression being significant for eight-week abdominal fat weights. In birds injected with gold thioglucose all weights were significantly depressed. In a separate investigation, histological evidence of generalised hypothalmic lesions was observed in both chicks (Gallus domesticus) and ducklings (Anas platyrhynchos) injected with bipiperidyl mustard or gold thioglucose. Similar lesions were absent in control chicks and ducklings. The possibility of the existence of a sex-steroid-mediated endorphinergic-glucostatic mechanism for appetite regulation in birds is discussed.     

Protective immunity against Newcastle disease: the role of cell-mediated immunity

The role of cell-mediated immunity (CMI) in protection of birds from Newcastle disease was investigated by two different strategies in which only Newcastle disease virus (NDV)-specific CMI was conveyed without neutralizing antibodies. In the first strategy, selected 3-wk-old specific-pathogen-free (SPF) birds were vaccinated with either live NDV (LNDV), ultraviolet-inactivated NDV (UVNDV), sodium dodecyl sulfate-treated NDV (SDSNDV), or phosphate-buffered saline (PBS) (negative control) by the subcutaneous route. Birds were booster vaccinated 2 wk later and challenged with the velogenic Texas GB strain of NDV 1 wk after booster. All vaccinated birds had specific CMI responses to NDV as measured by a blastogenesis microassay. NDV neutralizing (VN) and hemagglutination inhibition (HI) antibody responses were detected in birds vaccinated with LNDV and UVNDV. However, birds vaccinated with SDSNDV developed antibodies that were detected by western blot analysis but not by the VN or HI test. Protection from challenge was observed only in those birds that had VN or HI antibody response. That is, birds with demonstrable CMI and VN or HI antibody response were protected, whereas birds with demonstrable CMI but no VN or HI antibody response were not protected. In the second strategy, birds from SPF embryos were treated in ovo with cyclophosphamide (CY) to deplete immune cells. The birds were monitored and, at 2 wk of age, were selected for the presence of T-cell activity and the absence of B-cell activity. Birds that had a significant T-cell response, but not a B-cell response, were vaccinated with either LNDV, UVNDV, or PBS at 3 wk of age along with the corresponding CY-untreated control birds. The birds were booster vaccinated at 5 wk of age and were challenged with Texas GB strain of NDV at 6 wk of age. All birds vaccinated with LNDV or UVNDV had a specific CMI response to NDV, VN or HI NDV antibodies were detected in all CY-nontreated vaccinated birds and some of the CY-treated vaccinated birds that were found to have regenerated their B-cell function at 1 wk postbooster. The challenge results clearly revealed that CY-treated birds that had NDV-specific CMI and VN or HI antibody responses to LNDV or UVNDV were protected, as were the CY-nontreated vaccinated birds. However, birds that had NDV-specific CMI response but did not have VN or HI antibodies were not protected from challenge. The results from both strategies indicate that specific CMI to NDV by itself is not protective against virulent NDV challenge. The presence of VN or HI antibodies is necessary in providing protection from Newcastle disease.     

Effect of a single injection of adrenaline on shell ultrastructure in a series of eggs from domestic hens

1. Adrenaline (1 mg in 1 ml water) was administered subcutaneously to three hens to determine whether any changes in the shell ultrastructure of subsequent eggs would occur. 2. The egg shells were examined in a scanning electron microscope after plasma etching. 3. The first three eggs laid by each bird after adrenaline injection were compared with previously-collected normal eggs from the same birds and also with control eggs collected over the same period from three hens which had received no adrenaline. 4. The first egg laid following adrenaline treatment was essentially normal but both the second and third eggs showed severe structural disorganisation at all levels, from the mamillary caps up to the cuticular layer. 5. Eggs laid 20 d or more after adrenaline treatment had reverted to normal. 6. The findings suggest that the abnormal eggs laid after hens have been exposed to disturbance or stress are likely to be affected in not only their external appearance but also to be of poor structural quality.