Leukocyte numbers in hemorrhaged Japanese quail after microwave irradiation in ovo

1. Coturnix coturnix japonica eggs were exposed to 2.45-GHz continuous wave microwave radiation at an incident power density of 5 mW/cm2 (and a specific rate of 4 mW/g) during the first 12 days of embryogeny. After hatching, hematological changes in response to an acute hemorrhage were measured in exposed and nonexposed (control) juveniles of both sexes. 2. Exposure did not affect erythroid cell numbers either before or after hemorrhage. 3. Exposure affected the recovery of lymphocyte and heterophil numbers after hemorrhage, but the effect was sex-limited. 4. These data indicate that microwave irradiation during embryogeny in ovo affects the ability of Japanese quail to recover from an acute and voluminous hemorrhage and that these radiation effects are sex-limited and consistent with a previous report.     

Serum enzymes in hemorrhaged Japanese quail after microwave irradiation during embryogeny

1. Japanese quail eggs were exposed to 2.45 GHz continuous wave microwave radiation at an incident power density of 5 mW/cm2 and a specific absorption rate of 4.03 mW/g during the first 12 days of embryogeny. 2. After hatching, serum biochemical changes in response to hemorrhagic stress were measured following a hemorrhage of 30% of the calculated total blood volume. 3. Lactate dehydrogenase, beta-glucuronidase, acid phosphatase, glucose and protein were not affected by microwave irradiation during embryogeny either before or after hemorrhage. 4. Microwave irradiation in ovo affected the response of serum glutamic oxaloacetic transaminase activity to hemorrhagic stress in Japanese quail.     

Leukocyte numbers during the humoral and cell-mediated immune response of Japanese quail after microwave irradiation in ovo

1. Coturnix coturnix japonica eggs were exposed to 2.45-GHz continuous wave microwave radiation at an incident power density of 5 mW/cm2 (SAR = 4 mW/g) during the first 12 days of embryogeny. After hatching, leukocyte differential changes were measured in response to an injection with Alectoris graeca chukar red blood cells (CRBC) and in response to a phytohemagglutinin (PHA) injection in irradiated and nonirradiated (sham) quail of both sexes. 2. Microwave irradiation did not affect anti-CRBC hemagglutinin titers, PHA-evoked dermal swelling or leukocyte numbers and percentages. 3. In both the irradiated and sham irradiated males, lymphocyte percentages decreased while heterophil percentages increased after CRBC or PHA injection. 4. In ovo irradiation with microwaves did not alter the time course of either a humoral immune response or a cell-mediated immune response in Japanese quail.     

Hematological response of hemorrhaged Japanese quail after blood volume replacement with saline

1. Hematological responses to hemorrhage by phlebotomy with and without replacement of blood volume with saline were measured in juvenile and adult male Coturnix coturnix japonica. 2. Recovery of total peripheral erythrocyte numbers and total peripheral leukocyte numbers occurred within 72 hr postphlebotomy in both treatment groups. 3. Saline replacement of blood volume following hemorrhage increased the total numbers and differential percentages of circulating reticulocytes at 72 hr postphlebotomy above the reticulocyte values of phlebotomized quail receiving no saline in both adult and juvenile Japanese quail.     

Reproduction of Japanese quail after microwave irradiation (2.45 GHz CW) during embryogeny

Japanese quail (Coturnix coturnix japonica) embryos were irradiated continuously in ovo with 2.45-GHz continuous wave radiation during the first 12 days of embryogenesis at an incident power of 5 mW/cm2 and a specific absorption rate of 4.03 mW/g. The internal temperature of irradiated and nonirradiated (sham) eggs was 37.5 +/- 0.3 degrees C, which is the optimum temperature for incubating quail eggs. At 35 days after hatching irradiated and sham-irradiated males were paired with irradiated or sham-irradiated females and daily records of reproductive performance were collected through 224 days of age. Progeny were hatched from each of the male-female pairs, and progeny reproductive performance was measured from 35 through 168 days of age. Hatchability was not affected by irradiation during embryogeny. Mortality after hatching, egg production, egg weight, fertility, hatchability of eggs produced, and reproductive performance of the progeny were not affected by irradiation during embryogeny. These observations indicate that irradiation of quail embryos with low-level microwave radiation does not affect the reproductive capacity of the hatchlings or of progeny produced from quail irradiated during incubation.     

Glucocorticoid metabolism in the in ovo environment modulates exposure to maternal corticosterone in Japanese quail embryos (Coturnix japonica)

Maternal effects have gained attention as a method by which mothers may alter the physiological condition and phenotype of their offspring based upon current environmental conditions. The physiological and phenotypic outcomes of glucocorticoid-mediated maternal effects have been extensively studied in a variety of vertebrates; however, the underlying mechanism is currently unclear. Here, we injected tritiated corticosterone into the yolks of freshly laid Japanese quail eggs (Coturnix japonica) and traced its movement and metabolism through the in ovo development period. We found that corticosterone was extensively conjugated throughout the egg by the end of development, and while minimal corticosterone was detected within the embryo during development, accumulation of a conjugated metabolite in the embryo started to occur on day 6 of development. Because no movement and metabolism of corticosterone occurred in infertile eggs, our findings suggest that embryos are not passive recipients of maternal steroids, but instead appear to possess extensive metabolic capabilities, which may modulate their exposure to maternal steroids.     

Blood volume recovery in hemorrhaged japanese quail

• 1.1. Blood volume and plasma biochemical changes and feed and water consumption in response to a hemorrhage by phlebotomy of 30% of the calculated total blood volume with and without replacement of blood volume with physiological saline were determined in juvenile male Coturnix coturnix japonica.
• 2.2. Plasma protein and osmolality decreased rapidly posthemorrhage and did not recover by 72 hr posthemorrhage.
• 3.3. Plasma glucose, Na⁺ and K⁺ increased within Ihr postphlebotomy. Plasma Na⁺ returned to nonphlebotomized levels within 6 hr postphlebotomy.
• 4.4. Saline replacement of blood volume resulted in hypervolemia within 3–5 min postphlebotomy.
• 5.5. Phlebotomized quail receiving no saline recovered blood volume to 0 hr (nonphlebotomized) levels within l hr postphlebotomy.

     

Effect of in ovo leptin administration on the development of Japanese quail

Potential changes in the activity of endocrine axes related to growth as a result of leptin administration during embryonic development of birds were evaluated in the Japanese quail as a model bird with fast growth and development. On day 5 of incubation, 0.1 microg or 1 microg of recombinant mice leptin in 50 microl of phosphate buffered saline were injected into the albumen of eggs. Animals from each group were killed by decapitation on day 0, 2, 5, 7, 14, 21, 28, 35, 42, 49 and 56 of life. Plasma concentrations of triiodothyronine (T(3)), thyroxin (T(4)), corticosterone, testosterone, total lipids, triacylglycerols, cholesterol, glucose and alkaline phosphatase activity were measured. Quail treated by leptin hatched earlier (5-24 hours) and had a higher body weight than the control group (P<0.05-0.001). Mean body weight across the whole observed period was higher in both treated groups as compared to the control group (P<0.05). Leptin in ovo administration was accompanied by changes of endocrine and metabolic parameters during postembryonic development. The most prominent changes appeared immediately after hatching (T(3), T(4), total lipids, triacylglycerols) and before sexual maturity. It is suggested that leptin acts as a general signal of low energy status to neuroendocrine systems in birds which improves utilization of nutrients.     

Effects of corticosterone treatment on responses to fasting in Japanese quail

Plasma concentrations of corticosterone, a glucocorticoid hormone, have been reported to increase during fasting in some species of birds. Although Japanese quail are widely used in laboratory studies, corticosterone responses to fasting have not been described for this species. We therefore investigated the effects of 24 h of fasting on plasma corticosterone concentrations in quail. Previous work has shown that the corticosterone response to handling in quail may be affected by treatment with corticosterone, so we also measured corticosterone responses to 24 h of fasting in quail treated with corticosterone in their drinking water, and determined the effects of corticosterone on food intake immediately after a fast. Plasma corticosterone concentrations were unaffected by 24 h of fasting in controls and in three out of four groups of birds with varying corticosterone intakes. Fasting in birds can be divided into three phases, and our results suggest that quail remained in phase I during the 24 h fast. Food intake in the 3 h immediately after fasting was higher in four groups of birds with varying corticosterone intakes than in the control group, and was greatest in quail with the highest corticosterone intake. The current results suggest that corticosterone can stimulate food intake in quail following a period of food deprivation.     

Japanese quail selected for high plasma corticosterone response deposit high levels of corticosterone in their eggs

Poor habitat quality or body condition often correlates with high responsiveness of the hypothalamo-pituitary-adrenal (HPA) axis rather than with elevated baseline levels of glucocorticoids. We hypothesized that, for egg-laying vertebrates, high responsiveness of the HPA axis would correspond to high concentrations of corticosterone in yolk. We tested the prediction that Japanese quail (Coturnix coturnix japonica) selected for high plasma corticosterone response to brief immobilization (HS quail) would lay eggs with higher yolk corticosterone concentrations than birds selected for low response (LS quail). Quail from both lines were left undisturbed, outside of the stressors associated with daily management, before a first round of egg collection. In a second experiment, quail of both lines were experimentally stressed during the week before egg collection. In both cases we found quail from the HS line to lay eggs with significantly higher yolk corticosterone concentrations than quail of the LS line. After exposure to added experimental stressors, the line difference was more pronounced (increasing from 62% to 96%). There was no line difference in concentrations of yolk testosterone. Our results suggest that (1) genetic differences underly differences in the transfer of maternal corticosterone to yolk and (2) females may be able to control deposition of corticosterone into yolk through a mechanism independent of baseline corticosterone titers.     

Response of Japanese quail to hemorrhagic stress after exposure to microwave radiation during embryogeny

Coturnix coturnix japonica eggs were exposed to 2.45 GHz continuous wave microwave radiation at an incident power density of 5 mW/cm2 (SAR = 4 mW/g) during the first 12 days of embryogeny. After hatching, hematologic changes in response to an acute hemorrhage were measured in exposed and nonexposed (control) juveniles and adults of both sexes. Reticulocyte numbers and percentages were depressed below control numbers at 24 hr postphlebotomy in exposed adult females. Lymphocyte numbers were depressed below control levels at 24 hr postphlebotomy in exposed juvenile and adult males. At 72 hr heterophil numbers were depressed in exposed juvenile and adult males. These data suggest that microwave irradiation during embryogeny affects the ability of Japanese quail to recover from an acute and voluminous hemorrhage and that these radiation effects are small.     

Consequences of in ovo exposure to p,p'-DDE on reproductive development and function in Japanese quail

This study was conduced to assess the effects of a one time embryonic exposure to p,p'-DDE (dichlorodiphenyldichloroethylene; DDE) on the reproductive development and function in Japanese quail (Coturnix japonica). Embryos were exposed at day one of incubation to either 20 or 40 microg DDE or a sesame oil vehicle control (injection volume=20 microl). Onset of puberty, gonadal histopathology, sperm motility, cloacal gland size, and male copulatory behavior were assessed in adults. DDE accelerated onset of puberty in females and reduced male reproductive behaviors. Gonadal morphology and sperm motility appeared to be unaffected. Results from this study provide evidence that the neuroendocrine system may be more sensitive and less resilient to embryonic exposure to contaminants than traditional measures of reproductive success following contaminant exposure in adults. This study further supports the inclusion of behavioral assessments in toxicity tests.     

Effects of corticosterone treatment on growth, development, and the corticosterone response to handling in young Japanese quail (Coturnix coturnix japonica)

Corticosterone, a glucocorticoid secreted during stress responses, has a range of actions that help birds respond to stressors. Although effects of corticosterone treatment have been described in several avian species, the impacts of defined increases in plasma corticosterone on early development and on corticosterone stress responses are little known. These issues were addressed by providing quail with different doses of corticosterone in drinking water from days 8 to 38 post-hatch. The corticosterone dose consumed by each bird during treatment days 15-30 was calculated by measuring water intake. The corticosterone dose was inversely, but weakly, correlated with weights of the bursa, thymus, spleen, liver, testes, oviduct, muscle, and body, and positively correlated with peritoneal fat deposition. When birds were divided into groups based on their corticosterone intake, weights of the spleen, thymus, bursa, muscle, testes, and oviduct were significantly reduced in birds receiving the highest doses; with the exception of muscle, similar reductions were also observed in birds receiving medium doses, and thymic growth was inhibited in birds receiving low doses. The acute corticosterone stress response was measured by handling birds for 15 min. Plasma corticosterone was transiently increased at 15 min in control birds in response to the handling stressor. Some birds consuming low doses of corticosterone had corticosterone responses similar to control birds. Initial corticosterone concentrations were elevated in birds consuming higher doses of corticosterone. Plasma corticosterone in these birds decreased from 0 to 15 min, then increased from 15 to 30 min. The initial decrease could be due to corticosterone clearance, whilst the increase could indicate that the birds had a greater response than control birds to isolation as a stressor. Corticosterone treatment may have reduced the strength of corticosterone negative feedback within the hypothalamo-pituitary-adrenal axis. The results indicate that individuals and organs differ in their sensitivity to corticosterone. Moreover, elevated plasma corticosterone may disrupt the acute corticosterone stress response, and may thus reduce the ability of birds to cope with stressors.     

Infradian rhythms of the esophageal epithelium mitotic activity, corticosterone and thyroxin level in Japanese quail Coturnix japonica

Everyday dynamics of corticosterone and thyroxin levels on blood serum, as well as esophageal epithelium proliferation activity in Japanese quail males, have been investigated over a period of 16 days. It was estimated that different individuals synchronically exhibited a 4-day rhythm of the corticosterone serum level. The thyroxin level displayed a 3-day biorhythm. The mitotic index in the esophageal epithelium negatively correlated with the corticosterone level. The revealed infradian rhythms of the corticosterone and thyroxin levels, as well as esophageal epithelium mitotic index, should be taken into account while carrying out experiments concerning the determination of the hormone level and mitotic activity of epithelial tissues.     

Micronucleus induction after whole-body microwave irradiation of rats

Adult male Wistar rats were exposed for 2 h a day, 7 days a week for up to 30 days to continuous 2,450 MHz radiofrequency microwave (rf/MW) radiation at a power density of 5-10 mW/cm(2). Sham-exposed rats were used as controls. After ether anesthesia, experimental animals were euthanized on the final irradiation day for each treated group. Peripheral blood smears were examined for the extent of genotoxicity, as indicated by the presence of micronuclei in polychromatic erythrocytes (PCEs). The results for the time-course of PCEs indicated significant differences (P<0.05) for the 2nd, the 8th and the 15th day between control and treated subgroups of animals. Increased influx of immature erythrocytes into the peripheral circulation at the beginning of the experiment revealed that the proliferation and maturation of nucleated erythropoietic cells were affected by exposure to the 2,450 MHz radiofrequency radiation. Such findings are indicators of radiation effects on bone-marrow erythropoiesis and their subsequent effects in circulating red cells. The incidence of micronuclei/1,000 PCEs in peripheral blood was significantly increased (P<0.05) in the subgroup exposed to rf/MW radiation after eight irradiation treatments of 2 h each in comparison with the sham-exposed control group. It is likely that an adaptive mechanism, both in erythrocytopoiesis and genotoxicity appeared in the rat experimental model during the subchronic irradiation treatment.     

Micronucleus induction after whole-body microwave irradiation of rats

Adult male Wistar rats were exposed for 2 h a day, 7 days a week for up to 30 days to continuous 2450 MHz radiofrequency microwave (rf/MW) radiation at a power density of 5–10 mW/cm². Sham-exposed rats were used as controls. After ether anesthesia, experimental animals were euthanized on the final irradiation day for each treated group. Peripheral blood smears were examined for the extent of genotoxicity, as indicated by the presence of micronuclei in polychromatic erythrocytes (PCEs). The results for the time-course of PCEs indicated significant differences (P<0.05) for the 2nd, the 8th and the 15th day between control and treated subgroups of animals. Increased influx of immature erythrocytes into the peripheral circulation at the beginning of the experiment revealed that the proliferation and maturation of nucleated erythropoietic cells were affected by exposure to the 2450 MHz radiofrequency radiation. Such findings are indicators of radiation effects on bone-marrow erythropoiesis and their subsequent effects in circulating red cells. The incidence of micronuclei/1000 PCEs in peripheral blood was significantly increased (P<0.05) in the subgroup exposed to rf/MW radiation after eight irradiation treatments of 2 h each in comparison with the sham-exposed control group. It is likely that an adaptive mechanism, both in erythrocytopoiesis and genotoxicity appeared in the rat experimental model during the subchronic irradiation treatment.