Early embryonic administration of xenoestrogens alters vasotocin system and male sexual behavior of the Japanese quail

The copulatory behavior and the parvocellular vasotocin (VT) system of the nucleus of the stria terminalis (BST) are sexually dimorphic in the Japanese quail. Embryonic administration of estradiol benzoate (EB) induces an organizational effect determining the disappearance of such a dimorphism (male shows behavior and cerebral phenotype of the female). The VT parvocellular system can therefore be considered an accurate marker of the sexual differentiation of brain circuits and a very sensitive indicator of the activity of estrogen-like substances on neural circuits. To test this hypothesis we administered diethylstilbestrol (DES), a powerful synthetic xenoestrogen, genistein (GEN), a phytoestrogen produced by soy, and bisphenol A (BPA). After 3 days of incubation, quail eggs were injected with vehicle, EB, DES, GEN or BPA. Administration of BPA caused an early blockage of development and no further analyses were done on the BPA groups. At puberty, the copulatory behavior of EB- or DES-treated male quail was totally abolished, whereas only the highest doses of GEN determined a significant decrease of the behavior. After the tests, the animals were sacrificed and perfused. The fractional area (FA) covered by VT immunoreactivity was analyzed in BST, medial preoptic nucleus, and lateral septum by computerized image analysis. The FA was significantly reduced after treatment with EB, DES and GEN at high doses. These results confirm that the sexually dimorphic VT system of the Japanese quail is a sensible indicator of the effects of xenoestrogens at the level of the central nervous system.     

Mobility of Japanese quail spermatozoa and its relationship to egg fertility

Sperm mobility (SM) appears to be primary determinant of fertility in chicken and turkey. The aims of this study were to extend the concept to the Japanese quail by developing an assay to quantify SM, explaining the basis of SM using motility properties measured by CASA, and exploring the relationship between SM and egg fertility. The study was carried out in three stages: i) males (n = 20) and females (n = 20) were mated individually; ii) ejaculates were collected from 20 males, and SM was measured; iii) males (n = 20) and females (n = 20) were mated individually. In Stages I and III, data were collected for egg fertility, Sperm_OPVL and Holes_IPVL. In Stage II, SM assay was developed and assay conditions were defined: effect of sperm numbers on absorbance in Accudenz solution; effect of Accudenz concentration on sperm motility and mobility; effect of quail proctodeal gland foam extract and incubation temperature on SM at 37 and 41°C. The recorded absorbance of sperm movement was dependent on sperm numbers in the sperm suspension overlaying the Accudenz (p < .001). At 41°C, SM, progressively motile sperm, VCL, VSL and VAP were negatively affected by Accudenz concentration (p < .05). The effect of foam on SM and motility depended on an interaction between the concentration of foam extract and incubating temperature. Males were categorized into low, average and high SM phenotypes. These categories differed significantly (p < .001), but sperm motility and SM were not related to egg fertility. In conclusion, SM assay can be used to identify mobility phenotypes, but the poor relationship between SM and egg fertility indicates a need for further studies on interaction between the concentration of foam extract, incubating temperature, and in vivo sperm movement and egg fertilization success.     

Adaptability of Japanese quail chicks to conditions of simulated weightlessness

The objective of this study was to evaluate the adaptability of young Japanese quail chicks to the simulated weightlessness, represented by hypodynamy. Unsexed hatchlings were subjected to hypodynamy on either the first, second or third day of age and reared under these conditions to 21 days of age. During this period, the control quail chicks were housed in a floor box. The effect of hypodynamy on adaptability of chicks was significant (P < 0.001). Approximately 75% of all chicks exposed to hypodynamy were not able to adapt in three experimental groups, although significant differences in adaptability were not found between these groups. Those birds were considered as non-adapted (eliminated from experiment) that manifested hyperactivity, escape attempts, turning 180 degrees in the sling, soaking in the water from the drinker, as well as the total apathy, at least three times per day. This experiment confirmed that some quail chicks are capable of adapting to conditions simulating weightlessness to 21 days of age and that the first 2-weeks after hatching may be a critical period of quail sensitivity to hypodynamy. This finding raises a key issue relevant to rearing quails in simulated weightlessness until the age of sexual maturity.     

Relationship of serum alpha-amylase to aflatoxin resistance in Japanese quail

Serum alpha-amylase (S alpha A) activities were examined in a non-selected population (NS) of Japanese quail and in two lines genetically selected for resistance to aflatoxin. Normal S alpha A activities of both selected lines were significantly lower than that of the NS line (P less than or equal to 0.05). An oral dose of 2.5 mg aflatoxin/kg body weight significantly lowered S alpha A activity in NS quails after 24 hours; 18.0 mg/kg body weight was required to decrease the activity in selected lines to a similar degree. The two selected lines did not differ significantly in response. Disc tube gel electrophoresis of the serum revealed eight bands of S alpha A activity and six S alpha A zymogram phenotypes in the total population using an iodine-staining technique. Neither migration distance nor band width varied with genetic line, but the frequency of quails with at least one of three specific anodic bands was significantly higher in the selected lines (75% and 100%) than in the NS line (27%). The frequencies of the zymogram phenotypes with these bands were similarly increased in the resistant populations. Although there are differences in total S alpha A activity between the selected aflatoxin-resistant lines and the NS line, the presence of specific electrophoretic anodic band types and the S alpha A zymogram may be more useful indicators of aflatoxin resistance.     

Aflatoxin susceptibility of selected and control Japanese quail lines

1. The influence of aflatoxin on growth and mortality of diverse lines of quail was studied in five trials.

2. Feeding 5 or 10 /μg aflatoxin/g diet from 4 to 7 weeks of age resulted in significant decreases in body‐weight gain in all lines.

3. Similar gains for all treatments during the third week of feeding indicated that body weights were reduced only during the first two weeks.

4. Crop intubation with 4 or 8 mg aflatoxin/kg body weight resulted in different mortality patterns for the selected and control quail lines.     

Role of carbon dioxide and ion transport in the formation of sub-embryonic fluid by the blastoderm of the Japanese quail

1. The explanted blastoderm of the Japanese quail was used to explore the role of ions and carbon dioxide in determining the rate of sub-embryonic fluid (SEF) production between 54 and 72 h of incubation. 2. Amiloride, an inhibitor of Na+/H+ exchange, at concentrations of 10(-3) to 10(-6) M substantially decreased the rate of SEF production when added to the albumen culture medium. N-ethylmaleimide, an inhibitor of V type H+ ATPase, also decreased this rate but only to a small extent at the highest dose applied, 10(-3) M. Both inhibitors had no effect on SEF production when added to the SEF. 3. The inhibitors of cellular bicarbonate and chloride exchange, 4-acetamido-4'-isothiocyano-2,2'-disulphonic acid (SITS) and 4,4' diisothiocyanostilbene-2,2'-disulphonic acid (DIDS), had no effect upon SEF production. 4. Ouabain, an inhibitor of Na+/K+ ATPase, decreased SEF production substantially at all concentrations added to the SEF (10(-3) to 10(-6) M). Three sulphonamide inhibitors of carbonic anhydrase, acetazolamide, ethoxzolamide and benzolamide, decreased SEF production when added to the SEF at concentrations of 10(-3) to 10(-6) M. Benzolamide was by far the most potent. Neither ouabain nor the sulphonamides altered SEF production when added to the albumen culture medium. 5. Using a cobalt precipitation method, carbonic anhydrase activity was localised to the endodermal cells of the area vasculosa. The carbonic anhydrase activity was primarily associated with the lateral plasma membranes, which together with the potent inhibitory effect of benzolamide, suggests the carbonic anhydrase of these cells is the membrane-associated form, CA IV. 6. The changes in SEF composition produced by inhibitors were consistent with the production of SEF by local osmotic gradients. 7. It is concluded that a Na+/K+ ATPase is located on the basolateral membranes of the endodermal cells of the area vasculosa, and that a sodium ion/hydrogen ion exchanger is located on their apical surfaces. Protons for this exchanger would be provided by the hydration of CO2 catalysed by the membrane-associated carbonic anhydrase. Furthermore, it is proposed that the prime function of the endodermal cells of the area vasculosa is the production of SEF.     

Transport of radioiodide between blood and ovary of Japanese quail

Experimental data from blood and ovaries (oocytes) following injection of ¹²⁵Iodide were analyzed in terms of an open, two-compartmental model. Clearance of iodide by small, growing oocytes was 9.0 ul serum/mg/hour; 0.23 of the accumulated iodide left the oocytes per hour, presumably by diffusion. The accumulation of iodide by the small oocytes was shown to be saturable; K_m was 0.126 mg iodide/dl serum. These results support the concept that iodide accumulates in the small oocytes by active transport. FSH injected 2 hours prior to the ¹²⁵I caused a 50% increase in the clearance rate but no increase of the exit-rate constant.     

Utilization of free and protein-bound lysine in the Japanese quail

The utilization of dietary lysine for protein synthesis is affected by the digestibility of protein-bound lysine, by its intestinal resorption and by its oxidative catabolism. The approach chosen in this paper enables a comparison of the cumulative effect of these processes on the utilization of free and protein-bound lysine, respectively. The principle of the approach is based on a quantification of the expiration of 14C-labelled carbon dioxide after an oral administration of a diet, which contains L-(U-14C)-lysine either as a free amino acid or bound to yeast proteins. During an adaptation phase cockerels of the Japanese quail received a diet based mainly on ground wheat and wheat gluten. This diet was supplemented either with yeast proteins or with a mixture of L-amino acids which simulates the composition of the yeast proteins. In the main experiment the expiration of labelled carbon dioxide was measured during 240 minutes after the administration of the corresponding labelled diets. Just before treatment the animals were either in the postprandial phase or in a state of slight hunger. The maximum of expiration of labelled carbon dioxide occurred around the 60th minute after administration of the corresponding labelled diets. The cumulative expiration of labelled carbon dioxide, expressed in per cent of the radioactive dose used, amounts to 15.5% and 14.3% for free and protein-bound lysine, respectively. The utilization of both forms of lysine in the Japanese quail is lower than in broilers.     

The effect of water soluble trichloromonofluoromethane freon on the weight and proteins of the heart muscle in Japanese quail

An experiment was conducted to study the effects of water-soluble trichloromonofluoromethane freon (F-11, CC13F) on the weight of heart and on the level of freely extractable proteins in the heart of Japanese quail (Coturnix coturnix japonica). After three-week sublethal exposure of Japanese quail to the effect of freon the heart weight of the tested birds was found to have markedly decreased (however, the total body weight of the birds did not decrease). The tested quail also showed pronounced changes in the concentration of two protein fractions (an increase in one, a decline in the other), as determined by paper electrophoresis and polyacryl amide gel electrophoresis (however, the blood serum of the tested birds showed no changes in the levels of protein components).     

Divergent selection for shape of the growth curve in Japanese quail. 8. Effect of long-term selection on embryonic development and growth

1. Changes in embryonic development and growth were analysed in relation to direct changes in postnatal growth and correlated responses in egg parameters using Japanese quail lines selected for more than 30 generations for high (HG) and low (LG) relative gain of body weight (BW) between 11 and 28 d of age, and constant BW at 49 d of age.

2. During the first 42 h as well as at the end of incubation, LG embryos were developmentally accelerated in comparison with their HG counterparts. An expected increase of line divergence across generations was observed only in traits analysed at the end of incubation.

3. In contrast to early generations, LG embryos continuously exhibited a higher BW than HG embryos and this difference temporarily disappeared only around incubation d 8. Analogous to early generations, the HG compared with LG embryos showed two periods of transient growth retardation compensated subsequently by a higher growth rate (incubation d 3–8 and 8–16).

4. More pronounced growth retardation of HG versus LG embryos in late versus early generations corresponded to more distinct decrease of HG versus LG growth rate during the first post-hatch days. Likewise, a disappearance of line BW differences on incubation d 8 characterising the late generations corresponded to the elimination of line differences in adult BW.

5. Alterations of growth pattern were associated with changes of egg size. While HG quail maintained a relatively constant adult BW and egg size across generations, the gradually increasing incidence of large eggs in the LG line allowed selection of birds with higher growth potential, which in turn amplified the line differences in the embryonic BW and eliminated the line differences in adult BW. Line differences in egg composition (larger albumen with lower density in LG compared with HG eggs) apparently contributed to the strengthening of line developmental divergence during incubation.

6. Transient lack of nutrient supply to HG embryos due to their developmental delay is probably responsible for a higher HG versus LG embryo mortality.     

Calorimetric measurements of the energy and nitrogen metabolism of Japanese quail

1. Respiration calorimetry measurements were made over 4 d on normally‐fed groups of 10 male or 10 female quail during three periods commencing at 12, 19 and 26 d of age. Carcass compositions were determined on groups of birds of each sex at 2 and 5 weeks of age.

2. Expressed per unit body weight, food and metabolisable energy (ME) intake and weight gain decreased between 2 and 3 weeks of age, but remained constant thereafter.

3. No differences were observed between sexes for any variable measured. Efficiency of utilisation of ME for energy retention was only 0.33. This was mainly because the majority of energy was retained as protein tissue.

4. Nitrogen (N) retention (g/d) increased with increasing dietary N intake but when expressed as a proportion of N intake, declined from 0.46 at 2 weeks to 0.33 at 4 weeks. Fat retention increased substantially during week 4.

5. Carcass analyses showed that fat, protein and ash were higher for quail at 5 weeks than at 2 weeks of age. At 5 weeks carcass fat was only 60 g/kg.     

Inheritance of body weight and rate of gain in Japanese quail

1. The effects of hatch date, location and sex on the body weight and rate of gain in Coturnix coturnix japonica were examined.

2. The data conformed to a mixed model and adjustments were made for the fixed effects sex, hatch date and location. Adjustment improved the precision of the estimates by reducing the error variances significantly.

3. Rearing equal numbers of birds from each family at different locations made the interactions negligible.

4. Weight gain was greater if illumination intensity up to 35 d was less.

5. Females were not heavier on the 1st day but were heavier on the 14th and 35th days than males.

6. Variance components and genetic parameters were estimated.     

Maturation and ovulation of Japanese quail oocytes under in vitro conditions

1. An in vitro system for ovulation and maturation of Japanese quail oocytes is described.

2. Ovarian follicles removed from the ovary at 2, 4 or 6 h before the estimated time of ovulation may ovulate under in vitro conditions.

3. The presence of progesterone in the medium had a stimulatory effect on the process of maturation, as has been shown for Xenopus oocytes.     

Effect of restraint upon hypothalamic and adrenal catecholamines in Japanese quail

1. Changes in catecholamine concentration after periods of restraint up to 48 h in male Japanese quail have been measured. 2. Hypothalamic catecholamines decreased after 10 min restraint. Dopamine concentration increased after 4 h and persisted to 48 h. 3. Plasma catecholamines increased after 10 min, remained high for 2 h and then decreased despite continuing restraint. Catecholamine content of adrenals slightly decreased during restraint. 4. The activity of adrenal phenylethanolamine-N-methyl transferase increased from the 4th hour of restraint.     

Effects of photoperiod during incubation on embryonic and post-embryonic development of quail and chickens

The effect of light during incubation on embryonic and post‐embryonic development of White Leghorn chickens and Northern Bobwhite quail was determined. The treatments were: (a) 12 h light followed by 12 h dark (12L—12D), (b) 24 h light (24L—0D) and (c) 24 h dark (0L—24D) per day throughout incubation.

The development of the embryos of both species was accelerated by increasing the length of exposure to light during incubation. There were no differences in hatchability, weight at hatching, or at 4, 8, 12, 16 or 20 weeks of age. The incidence of healed navels was increased with greater length of exposure to light during incubation. No differences in egg production due to treatment were detected.