Dexamethasone alters the expression of genes related to the growth of skeletal muscle in chickens (Gallus gallus domesticus)

Glucocorticoids (GCs) are involved in the muscle wasting caused by trauma, inactivity, and stress. In the present study, three experiments were conducted to investigate the effect of GCs on the expression of genes related to muscle development in chickens. Broilers at 7 or 35 days of age were subjected to dexamethasone (DEX) treatment (2?mg/kg body mass (BM)) for 3 or 7 days. The expression levels of genes such as IGF1, IGF1 receptor, MSTN, WW domain containing E3 ubiquitin (UB) protein ligase 1, myogenic determining factor, and myogenic factor 5 were measured. The results showed that BM gain was significantly suppressed by DEX treatment. The plasma level of insulin was increased (P<0.05) by DEX treatment at feeding, whereas IGF1 was decreased (P<0.05). The expression of genes in the IGF1, myostatin, and UB-proteasome (UBP) pathways were altered by DEX treatment in age- and exposure time-related ways. These results suggest that GCs suppress IGF1 and upregulate myostatin and/or activated myostatin and the UBP pathway, which might be the source of the effect of GCs on muscle development.     

Muscle development in thyroidectomised chickens (Gallus domesticus)

The metabolic and contractile activity of muscle was determined in immature cockerels made hypothyroid by surgical thyroidectomy at 6 weeks of age. Four weeks after thyroidectomy the activity of Mg2+-activated myofibrillar ATPase and total phosphorylase was reduced in the fast-phasic, posterior latissimus dorsi (PLD) and scapulotriceps (ST) muscles. The activities of these enzymes were unaffected in the slow-tonic, anterior latissimus dorsi (ALD) muscle. Thyroidectomy had no effect on length of the muscles studied but reduced the weight of the ALD and ST muscles. These results suggest that hypothyroidism results in a "slowing down" of fast-phasic muscles, although it does not affect the activity of slow-tonic muscles.     

Ovarian aromatase activity in the domestic fowl (Gallus domesticus)

The effect of excessive tri-iodothyronine (T3) in vivo was assessed using normal human lymphocytes. Cells from normal subjects were frozen in liquid nitrogen before and after oral administration of T3 for 1 week to permit a direct comparison under identical culture conditions. Within the group of individuals studied, some subjects did show changes in B or T cell function but hypertri-iodothyroninaemia produced no consistent effect for the whole group on circulating T cell subsets or T and B cell activation measured by short-term culture or stimulation of lymphocyte cultures with phytohaemagglutinin or pokeweed mitogen. Tri-iodothyronine supplementation of cultures in vitro did not affect pokeweed mitogen stimulation. These findings suggest that the immunological abnormalities in Graves' disease are not the result of increased circulating thyroid hormone levels and that remission following medical treatment is due to an immuno-suppressive effect of the drug rather than the restoration of euthyroidism.     

Distinct features of dehydrocorticosterone reduction into corticosterone in the liver and duodenum of the domestic fowl (Gallus gallus domesticus)

The mammalian 11-beta hydroxysteroid dehydrogenase type 1 (11 betaHSD1) reduces glucocorticoids (GC) at C11 from the 11-keto-GC nonactive form to the 11-hydroxy-GC active form, an action essential for survival. Whereas GC metabolism at C11 and the role of 11 betaHSD1 are studied extensively in mammals, information about these in birds is scattered. Herein, we report the GC bidirectional metabolism in chickens. In hens' liver and duodenal mucosa, 11 betaHSD1-like mRNA expression was detected; and 11 betaHSD1-like immunoreactivity was found linked to membranes of hepatocytes and duodenal enterocytes. With either NADH or NADPH, the membranal fraction of liver and duodenal mucosa converted dehydrocorticosterone (A) into corticosterone (B) with K(m) (1.1-8.7 microM) and V(max) (10-40 pmol/mg protein/min) values similar to those reported for mammalian 11 betaHSD1. In the presence of NADP(+) or NAD(+), these membranal fractions oxidized B into A. With either NADPH or NADH, the cytosol of chicken liver and duodenal mucosa reduced A into B (K(m) of 1.1 - 2.3 microM and V(max) of 260-960 pmol/mg protein/min). These cytosolic fractions did not convert any amount of B into A when incubated with either NADP(+) or NAD(+). This may suggest that chicken liver and duodenal mucosa express 11 betaHSD1 that is a membrane-bound oxoreductase which uses both NADPH/NADP(+) and NADH/NAD(+) as cosubstrates. The substantial reduction of A into B (but no conversion of B into A) found in the cytosol is most likely executed by a unidirectional soluble reductase, different than 11 betaHSD1.     

Variation in virulence of West Nile virus strains for house sparrows (Passer domesticus)

The observation of avian mortality associated with West Nile virus (WNV) infection has become a hallmark epidemiologic feature in the recent emergence of this pathogen in Israel and North America. To determine if phenotypic differences exist among different WNV isolates, we exposed house sparrows (Passer domesticus) to low passage, lineage 1 WNV strains from North America (NY99), Kenya (KEN), and Australia (KUN; also known as Kunjin virus). House sparrows inoculated with the NY99 and KEN strains experienced similar mortality rates and viremia profiles. The KUN strain elicited significantly lower-titered viremia when compared with the other strains and induced no mortality. This study suggests that natural mortality in house sparrows due to Old World strains of WNV may be occurring where the KEN strain occurs.     

Serologic diagnosis of West Nile and St. Louis encephalitis virus infections in domestic chickens

Adult domestic chickens were infected with West Nile virus (WNV) or St. Louis encephalitis virus (SLEV) and challenged with homologous or heterologous virus at 21 or 56 days postinfection (dpi). Sera were collected at selected time points after infection and assayed by enzyme immunoassay (EIA), plaque reduction neutralization test (PRNT), and a Western blot (WB) alternative to PRNT. EIA results were sensitive and accurate (few false positives) but not specific, requiring a confirmatory test to determine virus infection history. PRNT results generally were specific until challenge, after which test results were frequently equivocal and inadequate to determine first or second infecting virus. WB results confirmed the serologic cross-reactivity between WNV and SLEV envelope protein. Non-structural protein 1 and pre-membrane protein reactivities were highly specific for WNV during SLEV infection, but less specific for SLEV during WNV infection. WB and PRNT specificities were similar for both viruses from 6 to 14 dpi, and sensitivities to WNV were virtually identical.     

Dietary protein restriction stress in the domestic chicken (Gallus gallus domesticus) induces remodeling of adrenal steroidogenic tissue that supports hyperfunction

The stress of dietary protein restriction in the immature domestic turkey (Meleagris gallopavo) induces adrenal steroidogenic hypofunction that is associated with an alteration in the proportion of density-dependent subpopulations of steroidogenic cells within the adrenal gland. In contrast, when imposed on immature chickens, this nutritional stressor induces long-term enhancement of adrenal steroidogenic function. However, whether this alteration in function is accompanied by a remodeling of chicken adrenal steroidogenic tissue as in the turkey is not known. To address this question, immature cockerels (2 weeks old) were fed established isocaloric synthetic diets containing either 20% (control) or 8% (restriction) soy protein for 4 weeks. Adrenal glands were processed for the isolation of defined, density-separable, adrenal steroidogenic cell subpopulations: three low-density adrenal steroidogenic cell subpopulations [LDAC-1 (rho = 1.0285-1.0430 g/ml), LDAC-2 (rho = 1. 0430-1.0485 g/ml), and LDAC-3 (rho = 1.0485-1.0500 g/ml)] and one high-density subpopulation [HDAC (rho = 1.0510-1.0840 g/ml)]. The steroidogenic function of these cell subpopulations was assessed. Protein restriction consistently, but differentially, enhanced maximal ACTH-induced corticosterone production by the subpopulations: values of LDAC-1, -2, and -3 and HDAC from protein restricted birds were, respectively, 116, 43, 33, and 20% greater than those of corresponding cell subpopulations from control birds. However, it had contrasting influences on maximal ACTH-induced aldosterone production by the cell subpopulations. Whereas the value of LDAC-1 from protein-restricted birds was 70% greater than that from control birds, the values for LDAC-2 and -3 were not different from those of the control, and the value for HDAC was 22% less than that of the control. Protein restriction also altered the cell subpopulation composition of the adrenal gland: compared to control, it increased the proportion of LDAC-1 by 46% and decreased the proportion of LDAC-3 and HDAC by 34 and 20%, respectively. Thus, dietary protein restriction increased the proportion of cells (i.e., LDAC-1) having the greatest enhancement in corticosteroid production. This pattern of remodeling of chicken adrenal steroidogenic tissue in response to dietary protein restriction contrasts sharply with the pattern that occurs in another galliform species, the domestic turkey.     

West Nile virus: immunity and pathogenesis

West Nile virus (WNV) is a neurotropic, arthropod-borne flavivirus that is maintained in an enzootic cycle between mosquitoes and birds, but can also infect and cause disease in horses and humans. WNV is endemic in parts of Africa, Europe, the Middle East, and Asia, and since 1999 has spread to North America, Mexico, South America, and the Caribbean. WNV infects the central nervous system (CNS) and can cause severe disease in a small minority of infected humans, mostly immunocompromised or the elderly. This review discusses some of the mechanisms by which the immune system can limit dissemination of WNV infection and elaborates on the mechanisms involved in pathogenesis. Reasons for susceptibility to WNV-associated neuroinvasive disease in less than 1% of cases remain unexplained, but one favored hypothesis is that the involvement of the CNS is associated with a weak immune response allowing robust WNV replication in the periphery and spread of the virus to the CNS.     

Thyroid hormones inhibit growth hormone secretion in domestic fowl (Gallus domesticus)

Daily blood samples from four mares were assayed for cortisol through a total of eight ovulatory cycles. Mean cortisol concentrations on days -14, -13, -10, -9 and -8 before ovulation (dioestrus) were greater than on days -5 to -1 (oestrus). The highest mean (+/- S.E.M) value of cortisol occurred on day -10 (260 +/-28 nmol/l) and the lowest on day -2 (142 +/- 14 nmol/l). A single episode on a day in late dioestrus characterized the maximum cortisol value per cycle for five of eight cycles. Extraction of plasma samples with petroleum ether or chromatography before assay, to eliminate interference from progesterone and its metabolites, did not alter the pattern of high dioestrous and low oestrous cortisol concentrations. Maximum follicular diameter at ovulation was negatively correlated with mean cortisol concentration for that cycle. These results indicate that in the mare the adrenals secrete cortisol more actively during dioestrus than during oestrus and suggest that a decline in cortisol values at oestrus may favour full follicular growth and ovulation.     

Dietary protein restriction stress in the domestic fowl (Gallus gallus domesticus) alters adrenocorticotropin-transmembranous signaling and corticosterone negative feedback in adrenal steroidogenic cells

Previous work with growing chickens (Gallus gallus domesticus) indicates that transient dietary protein restriction induces long-term enhancement of adrenal steroidogenic function in response to adrenocorticotropin (ACTH). The present study investigated two possible cellular functions mediating this enhanced response: (a) ACTH signal transduction and dissemination and (b) short-loop feedback inhibition of ACTH-induced corticosterone production by exogenous corticosterone. Cockerels (2 weeks old) were fed isocaloric synthetic diets containing either 20% (control) or 8% (restriction) soy protein for 4 weeks. Adrenal glands were processed for the isolation of adrenal steroidogenic cells nearly devoid of chromaffin cells ( approximately 90% adrenal steroidogenic cells). Results of experiments to assess signal transduction and dissemination indicated that protein restriction selectively enhanced ACTH-induced corticosterone production mediated by the cyclic AMP (cAMP)-dependent pathway. In addition, protein restriction substantially counteracted exogenous corticosterone-dependent inhibition of acute ACTH-induced corticosterone production (by 40.7% vs control). The proximal portion of the cAMP pathway seemed most affected by this stressor. Protein-restricted cells exhibited enhanced homologous sensitization to ACTH (136% greater than that of control cells) which appeared to be localized at a step(s) prior to or at the formation to cAMP. Also, maximal ACTH-induced cAMP production and sensitivity to ACTH in terms of cAMP production by protein-restricted cells were, respectively, 2.2 and 15.8 times those of control cells. However, variable results were obtained from other experiments designed to pinpoint the altered early steps in ACTH-transmembranous signaling. For example, with intact cells, cAMP responses to cholera toxin (CT) and forskolin (FSK) did not corroborate the results suggesting an augmentation of ACTH-signal transduction induced by protein restriction. Furthermore, basal and stimulatable (by ACTH, CT, FSK, and NaF) adenylyl cyclase activities from membranes from protein-restricted cells were, respectively, 47.2 and 40.2% less than those from control cells (normalized to 10(7) cell equivalents of crude membranes). Collectively, these findings suggest that protein restriction stress potentiates ACTH-induced corticosterone secretion by chicken adrenal steroidogenic cells in at least two ways: (1) on the proximal end, by modulating unknown factors which enhance cellular sensitivity to ACTH, ACTH receptor-adenylyl cyclase coupling, and adenylyl cyclase activity, and (2) on the distal end, by suppressing end-product corticosterone negative feedback, thus facilitating an increase in net corticosterone secretion.     

A comparison of variations in plasma luteinizing hormone concentrations in male and female domestic chickens (Gallus domesticus) from hatch to sexual maturity.

Changes in plasma LH concentrations were followed in chickens of both sexes from hatch to sexual maturity using a radioimmunoassay. Mean levels of LH were lower in the females than in the males at all stages of development. These levels rose rapidly in both sexes during the first week after hatch to maxima of 6-5 +/- 1-2 (S.E.M.) ng/ml (n = 6) in the males and 4-6 +/- 0-6 ng/ml (n = 6) in the females. Thereafter levels of the hormone in the circulation stabilized in the males but fell over a period of 1 or 2 weeks in the females to 2-5-3 ng/ml. Plasma LH levels started to rise steeply in both sexes when they were between 16 and 19 weeks old at the same time as there was an increase in the rate of comb growth. Afterwards in six of the males studied in detail the mean plasma LH level rose significantly (P less than 0-01) over a period of 5-8 weeks from 8-1 +/- 1-2 to 13-2 +/- 1-9 ng/ml. In a parallel study on six females the rate of LH secretion increased for approximately 3 weeks and then decreased for about the same period forming a prepubertal LH peak. The first eggs were laid between 22 and 25 weeks of age when mean plasma LH levels had fallen to about 1-8 ng/ml. The mean plasma LH level in these hens when they were laying (1-8 +/- 0-3 ng/ml) was significantly lower (P less than 0-01) than when they were sexually immature (2-7 +/- 0-3 ng/ml). The duration of the period of rapid comb growth in each bird was closely related in the males to the time during which prepubertal LH peak. Differences in mean plasma LH concentrations in individual birds of either sex before the onset of puberty appeared to be related to subsequent reproductive performance.     

Daily infusion of corticosterone and reproductive function in the domestic hen (Gallus domesticus)

The purpose of this study was to examine the effect of the daily infusion of corticosterone on reproductive function in the laying hen and to determine the relationship between the cyclic pattern of plasma concentrations of corticosterone on the open-period for the preovulatory release of LH. An exogenous rhythm of plasma levels of corticosterone was generated using an osmotic pump. Corticosterone was infused subcutaneously into laying hens at rates of 5, 10, 15 or 30 micrograms/hr for a duration of 10 hr beginning with the onset of darkness or at 15 micrograms/hr for 4 hr, or continuously at 30 micrograms/hr. Daily infusions greater than 15 micrograms/hr and the continuous infusion resulted in cessation of ovulation, ovarian and oviductal regression, hyperphagia, and elevated levels of plasma corticosterone compared to that observed in control hens. The hens which were infused with 5 or 10 micrograms/hr of corticosterone maintained normal reproductive function with plasma concentrations of corticosterone that were approximately the same as those in the control hens. The effect of infusing 10 micrograms/hr of corticosterone on the open-period for the preovulatory release of LH was determined under constant light. No significant changes were observed in the frequency distribution of the times of oviposition when hens were infused with 10 micrograms/hr of corticosterone for 12 hr from 9:00 to 21:00 hr or 21:00 to 9:00 hr each day.(ABSTRACT TRUNCATED AT 250 WORDS)     

Sodium-dependent intracellular pH regulation in granulosa cells of the domestic hen (Gallus domesticus)

The existence and importance of the Na+/H+ exchanger in intracellular pH (pHi) regulation in ovarian cells was studied in acid-loaded avian granulosa cells by monitoring the recovery of normal pHi using a trapped fluorescein derivative as an indicator. The resting pHi of freshly isolated granulosa cells from preovulatory follicles was 6.80 +/- 0.08 when the extracellular pH (pHo) and sodium concentration (Na+o) were 7.3 and 144 mmol/l respectively. While exposure of granulosa cells to high pHo (pHo greater than 7.45) medium shifted the pHi upward with time, incubation of the cells in low pHo (pH less than 6.80) buffer resulted in a significant decrease in pHi. In contrast, pHi remained constant when pHo was varied between the broad range of 6.8-7.4. When the cytoplasm was acidified by treatment with nigericin in choline+ buffer, both the magnitude and rate of recovery of normal pHi was suppressed significantly with decreasing pHo, but increased in high pHo medium. The recovery of pHi was dependent upon the concentration of extracellular sodium, in that the recovery rate and magnitude increased concomitantly with increases in Na+o concentrations, while the recovery was abolished when Na+o was completely replaced with choline+. In addition, the sodium ionophore monensin enhanced the recovery rate of normal pHi in a concentration-dependent manner. This action of monensin was observed only when sodium was present in the incubation medium, indicating that Na+o entry is important for the recovery of normal pHi. Monensin also evoked further cytoplasmic alkalinization in fully recovered cells, with a relative net effect dependent upon the level of Na+o present.(ABSTRACT TRUNCATED AT 250 WORDS)     

Diurnal variation in plasma luteinising hormone levels in the domestic fowl (Gallus domesticus)

Variations in the concentration of luteinising hormone (LH) in the blood were measured over 24 hr in sexually immature chickens maintained on long daily photoperiods of 18L:6D, 16L:8D, or 14L:10D. A significant increase in the concentration of LH occurred shortly after the onset of the dark period in both sexes. The possible significance of this observation in relation to neural control of the ovulatory cycle is discussed.     

West Nile virus circulation detected in northern Italy in sentinel chickens

Ninety percent (56/62) of sentinel chickens introduced to two regions within the Italian Alps seroconverted to West Nile virus (WNV) during the summer of 2005, showing a range of antibody titres from 1/20 to 1/320 in a virus neutralization test. Neutralization specificity for WNV antibodies was confirmed on an additional 34 sera that were tested in parallel against WNV (16/34 seropositivity), Usutu virus (3/34 seropositivity) and Koutango virus. The geometric mean neutralizing titre (GMT) calculated for WN-specific antibodies was 33.68 and did not differ significantly amongst sample sites, although the overall results indicate more active circulation of WNV at the higher elevations. Such high levels of seroconversion raise the possibility that many chickens may have been exposed to virus via routes other than mosquito transmission. No chickens or any other local animals were associated with illness due to WNV implying that WNV, and to a much lower extent Usutu virus, circulate harmlessly amongst wildlife species in northern Italy from late May onwards until early autumn.     

Influence of pinealectomy on plasma and extrapineal melatonin rhythms in young chickens (Gallus domesticus)

A specific radioimmunoassay was validated for the quantitative measurement of melatonin (MT) levels in plasma and homogenates of the pineal gland, Harderian gland, or retinae of young chickens. Single-comb White Leghorn (SCWL) cockerels were raised under a 12L:12D light/dark cycle for two experiments. In Experiment 1, 12 birds were bled and immediately killed for their pineal glands at 4-hr intervals during a single light/dark cycle at 25 days of age (25 da) for simultaneous determination of changes in MT levels in the plasma and pineal gland. Plasma MT levels were low during photophase (100 pg/ml) and reached a distinct peak (390 pg/ml) at mid-scotophase. A parallel MT rhythm was found in pineal homogenates where the average MT content during scotophase (7.4 ng/gland) was 10 times higher than the average MT content of pineal glands obtained during photophase. In Experiment 2, SCWL cockerels were either pinealectomized or sham-operated (PN) at 8 to 10 da. At 25 da, six birds from each surgical treatment group, including unoperated controls (C), were bled at 4-hr intervals, corresponding to those in Experiment 1, during a single light/dark cycle. Immediately after being bled, each bird was killed and the eyes and Harderian glands were removed for measurement of their MT contents. Pinealectomy completely abolished the plasma MT rhythm which in intact chicks (PN and C) reached a sharp peak (298 pg/ml) at mid-scotophase. Although not affected by surgical treatment, retinal MT levels showed a higher amplitude rhythm with a prominent peak (4 ng/retina) at mid-scotophase that was 15 times higher than the average retinal MT content during photophase. A modest nocturnal MT rhythm was found in the Harderian gland where the average MT level for all surgical treatment groups during scotophase (89 pg/100 mg wet wt) was only 51% higher than that observed for photophase. These data indicate that the plasma MT rhythm in chickens is derived solely from MT secreted into blood by the pineal gland and that the extrapineal MT produced rhythmically in both the retina and Harderian gland does not contribute to the plasma MT rhythm.     

Rapid active assay for the detection of antibodies to West Nile virus in chickens

To reduce the assay time for detecting virus-specific antibodies in serum, we developed microarray-based active immunoassay techniques for detecting West Nile virus (WNV)-specific IgM molecules in chicken blood. The assay uses electrophoretic concentration of IgM molecules onto WNV antigens arrayed on a dialysis membrane followed by detection of bound IgM molecules with functionalized magnetic beads as active labels. This assay takes only 15 minutes and has the same sensitivity as a commercially available human WNV IgM antibody-capture enzyme-linked immunosorbent assay (commonly called a MAC-ELISA) modified for use with chicken sera.