Egg yolk and serum antibody titers of broiler breeder hens immunized with uricase and or urease.

This study evaluated whether broiler breeder hens immunized with uricase (UC), urease (UE), or UC + UE would develop antibody (IgY) titers against these enzymes to prevent manure-N degradation and NH(3) release. Ross × Arbor Acres hens were assigned to PBS (control), UC, UE, or UC + UE injection treatments. Each group had 19 hens per treatment. On d 0, each of the enzymes or PBS was emulsified with complete Freund's adjuvant and administered intramuscularly, whereas on d 7 and 14, a booster injection of PBS or enzymes was administered as an incomplete adjuvant. Blood samples were taken on d 0, 4, 9, 12, 17, 21, and 24 for serum-specific IgY titer analysis. Eggs were collected for yolk-specific IgY titer analysis. Manure samples were taken for nutrient, pH, and NH(3) measurements. Elevated egg yolk anti-UC-IgY titers were observed from UC-immunized hens after the second immunization (P ≤ 0.0001), and they remained higher than those of the PBS- or UE-immunized hens from d 9 to 24. After the first injection, egg yolk anti-UE-IgY titers from hens immunized with UE or the combined antigen were greater than those of birds injected with PBS or UC (P ≤ 0.01). The serum anti-UC-IgY response to UC immunization was observed after the first injection (P ≤ 0.01) and on d 9 (P ≤ 0.0001), and titers remained greater than those of hens immunized with PBS or UE until d 28. The serum anti-UE-IgY titers remained low until much later compared with the anti-UC-IgY titers. Only at 24 and 28 d were anti-UE-IgY titers significantly greater in the UE-immunized hens than in hens immunized with PBS or UC. Hens immunized with UC or UE responded with both egg yolk and serum IgY titers. The combined antigens were significantly greater than the PBS control but had less effect than the individual UC or UE in both the egg yolk and serum. These findings indicate that despite measurable egg yolk and serum IgY titers, immunizing hens with UC, UE, or the combined antigens did not affect the manure nutrients or NH(3) emissions of the treated hens.

Gonadotrophin-inhibitory hormone receptor expression in the chicken pituitary gland: potential influence of sexual maturation and ovarian steroids.

Gonadotrophin-inhibitory hormone (GnIH), a hypothalamic RFamide, has been found to inhibit gonadotrophin secretion from the anterior pituitary gland originally in birds and, subsequently, in mammalian species. The gene encoding a transmembrane receptor for GnIH (GnIHR) was recently identified in the brain, pituitary gland and gonads of song bird, chicken and Japanese quail. The objectives of the present study are to characterise the expression of GnIHR mRNA and protein in the chicken pituitary gland, and to determine whether sexual maturation and gonadal steroids influence pituitary GnIHR mRNA abundance. GnIHR mRNA quantity was found to be significantly higher in diencephalon compared to either anterior pituitary gland or ovaries. GnIHR mRNA quantity was significantly higher in the pituitaries of sexually immature chickens relative to sexually mature chickens. Oestradiol or a combination of oestradiol and progesterone treatment caused a significant decrease in pituitary GnIHR mRNA quantity relative to vehicle controls. GnIHR-immunoreactive (ir) cells were identified in the chicken pituitary gland cephalic and caudal lobes. Furthermore, GnIHR-ir cells were found to be colocalised with luteinising hormone (LH)beta mRNA-, or follicle-stimulating hormone (FSH)beta mRNA-containing cells. GnIH treatment significantly decreased LH release from anterior pituitary gland slices collected from sexually immature, but not from sexually mature chickens. Taken together, GnIHR gene expression is possibly down regulated in response to a surge in circulating oestradiol and progesterone levels as the chicken undergoes sexual maturation to allow gonadotrophin secretion. Furthermore, GnIHR protein expressed in FSHbeta or LHbeta mRNA-containing cells is likely to mediate the inhibitory effect of GnIH on LH and FSH secretion.

Effect of heat stress on production parameters and immune responses of commercial laying hens.

The present study was conducted to determine the adverse effects of high temperature and humidity not only on live performance and egg quality but also on immune function in commercial laying hens. One hundred eighty 31-wk-old laying hens at peak production were used in this study. Hens were housed in cages (15 cages of 4 birds/cage) in each of 3 environmental chambers and received 1 of 3 treatments. The 3 treatments were control (average temperature and relative humidity), cyclic (daily cyclic temperature and humidity), and heat stress (constant heat and humidity) for 5 wk. Different production and immune parameters were measured. Body weight and feed consumption were significantly reduced in hens in the heat stress group. Egg production, egg weight, shell weight, shell thickness, and specific gravity were significantly inhibited among hens in the heat stress group. Likewise, total white blood cell (WBC) counts and antibody production were significantly inhibited in hens in the heat stress group. In addition, mortality was higher in the heat stress group compared to the cyclic and control groups. Even though T- and B-lymphocyte activities were not significantly affected by any of the treatments, lymphocytes from hens in the heat stress group had the least activity at 1 wk following treatment. These results indicate that heat stress not only adversely affects production performance but also inhibits immune function.

Initiation of humoral immunity. I. The role of cytokines and hormones in the initiation of humoral immunity using T-independent and T-dependent antigens.

The early events during the initiation of immune responses following the injection of T-independent (lipopolysaccharide, LPS) and T-dependent (bovine serum albumin, BSA) antigens were studied in immature male chickens. Specifically, the role of cytokines and hormones in the initiation of humoral immunity against these antigens was investigated. Both interleukin-1 (IL-1) and tumor necrosis factor (TNF-alpha) increased significantly post-LPS but not post-BSA injection. While interleukin-2 (IL-2) significantly decreased post-LPS injection, IL-2 significantly increased post-BSA injection. Furthermore, corticosterone levels significantly increased and tri-iodothyronine (T(3)) levels significantly decreased post-LPS but not post-BSA injection. In this study, the results indicate that although LPS and BSA can induce a humoral antibody response in chickens, they activate different cytokines and neuroendocrine network systems.

Initiation of humoral immunity. II. The effects of T-independent and T-dependent antigens on the distribution of lymphocyte populations.

The effect of injecting T-independent (lipopolysaccharide, LPS) and T-dependent (bovine serum albumin, BSA) antigens on the redistribution of lymphocyte populations in immature male chickens was investigated. In the blood, percentages of total T-cells (CD3+), T-helper cells (CD4+), and T-cytotoxic/suppressor cells (CD8+) significantly decreased post-LPS injection (PLI) but not post-BSA injection (PBI), while percentages of monocytes/thrombocytes (K1+) significantly increased PLI. Interleukin-1 receptor expression on blood lymphocytes increased significantly PLI and PBI. In the spleen, the percentages of total T-cells (CD3+) increased significantly PLI and PBI, macrophage (K1+) percentages increased significantly PLI, while B-cell percentages decreased significantly PLI. These results indicate that following antigen injection, there is a redistribution of peripheral blood lymphocytes (specifically T-lymphocytes) to secondary lymphoid organs and the kinetics and magnitude of the changes can differ according to the type of antigen used.

Melatonin and the enhancement of immune responses in immature male chickens.

Understanding the role of melatonin in affecting different physiological functions, especially immune responses, is becoming increasingly important in the basic and applied sciences. Enhancing the immune response will result in increasing disease resistance and, therefore, improve production efficiency. The purpose of the present study was to investigate the effects of melatonin, administered during the light or dark period, on BW, feed consumption (FC), and immune responses of immature chickens. Eight-week-old Cornell White Leghorn males were used in this study. The doses of melatonin were 0, 5, 10, 20, and 40 mg/kg BW. Melatonin was administered s.c. every 24 h for 7 consecutive d. The chicks were randomly divided into two groups; one group received injection during the middle of the light period, and the other group received injection during the middle of the dark period. All birds received 16 h light and 8 h darkness during a 24-h period. Body weights were measured before and after melatonin treatment, and FC was also measured. After the seven injections, blood samples were collected from the brachial vein, and total white blood cell (WBC) counts, differential cell counts, and activities of T and B lymphocytes were measured. Body weight was not significantly affected by dose of melatonin or time of injection. Furthermore, melatonin did not significantly affect FC; however, FC was significantly lower in the group that was injected in the dark vs. light period. The WBC counts of birds injected with 40 mg melatonin/kg BW were significantly higher than the WBC counts of saline-injected birds. The heterophil/lymphocyte (H/L) ratios of birds injected during the light period were significantly higher than those of birds injected during the dark period. T- and B-lymphocyte proliferation were significantly higher in birds injected with 40 mg melatonin/kg BW compared to saline-injected birds. These results indicate that melatonin in vivo is important in enhancing not only circulating WBC but also activities of B and T lymphocytes of immature male chickens without adversely affecting BW.

Effects of corticotropin releasing factor on the production of adrenocorticotropic hormone by leukocyte populations.

1. Corticotropin-releasing factor (CRF), a neuropeptide with immunomodulating properties, is known to stimulate avian splenic leukocytes to produce adrenocorticotropic hormone (ACTH). 2. The present study was to determine which avian splenic leukocyte subpopulation(s) produce ACTH in response to CRF stimulation. 3. Splenic leukocytes from 8-week-old male chickens were isolated on Histopaque 1077 and macrophages were separated from lymphocytes by adherence to a polystyrene surface. 4. Different concentrations of CRF (0, 5, 50, 500 or 1000 ng/m) were incubated with the different leukocyte populations, supernatants were collected and ACTH was measured using a radioimmunoassay. 5. Isolated macrophages, stimulated with CRF, produced significantly more ACTH than either unstimulated macrophages or CRF-stimulated lymphocytes, suggesting that ACTH may be produced by a particular subset of leukocytes, the macrophages (and monocytes), in response to CRF stimulation.

Effects of corticosterone in vivo and in vitro on adrenocorticotropic hormone production by corticotropin releasing factor-stimulated leukocytes.

Corticotropin releasing factor (CRF) and corticosterone have been shown to affect immune cell function. Previously, we have shown that CRF stimulates immunoreactive adrenocorticotropic hormone (ACTH) production by leukocytes. In this study, splenic leukocytes from corticosterone-injected chickens failed to show a CRF-induced increase in ACTH production. In addition, corticosterone in vitro inhibited the production of leukocyte ACTH as well as the stimulatory effect of CRF on splenic leukocyte ACTH production. These findings show that, as with anterior pituitary ACTH production, CRF-stimulated leukocyte ACTH production is inhibited by glucocorticoids.

Validation of an assay to measure adrenocorticotropin in plasma and from chicken leukocytes.

A RIA for mammalian adrenocorticotropic hormone (ACTH) was modified and validated to measure chicken ACTH. The assay was capable of detecting an increase in chicken plasma ACTH following treatments known to increase plasma ACTH. Both splenic and peripheral blood leukocytes stimulated with corticotropin-releasing factor (CRF) showed a significant increase in ACTH production compared with unstimulated leukocytes. This finding supports the conclusion that the substance produced by leukocytes previously shown in our laboratory to stimulate adrenal cells to secrete corticosterone is immunoreactive ACTH.

The endocrine function of the immune cells in the initiation of humoral immunity.

The endocrine functions of different hormones are well documented. Recently, however, evidence of the immune function of several hormones has been accumulated. These findings raised the possibility that immune cells might secrete hormones and in turn self-regulate different immune functions. Indeed, immune cells have been found to secrete different peptide and protein hormones. Adrenocorticotropin hormone (ACTH) is the most studied and was found to be secreted by lymphocytes. In the present authors' laboratory, it was found that not only do lymphocytes secrete ACTH but also that circulating concentrations of corticosterone increased following antigen challenge. It was also observed that at the time of the increased corticosterone, there was a redistribution of different lymphocyte subpopulations from the blood to spleen, the site of antigen presentation and antibody production. It was concluded that the endocrine function of lymphocytes might be important in the initiation of antibody production in chickens.

Ovine corticotropin-releasing factor increases endocrine and immunologic activity of avian leukocytes in vitro.

Treatment of splenic leukocytes from Cornell K strain male chickens (homozygous at the B15 locus of the major histocompatibility complex) with ovine corticotropin-releasing factor (oCRF), before their co-incubation with naive chicken adrenal cells, resulted in an increase in corticosterone production. Supernatants from the oCRF-treated splenic leukocytes caused a time-dependent increase in corticosterone production when incubated with chicken adrenal cells. Adding oCRF directly to chicken adrenal cells did not increase corticosterone production. Pretreatment of peripheral leukocytes with oCRF increased their activity in a concanavalin A mitogen assay. Thus, chicken leukocytes stimulated with corticotropin releasing factor appear to increase the production of an "adrenocorticotropin-like" substance (adrenocorticotropin-like because it increases corticosterone production by adrenal cells), and increased their cell-mediated immune activity.