Antigenic mapping of the hemagglutinin of the H9 subtype influenza A viruses using sera from Japanese quail (Coturnix c. japonica).

IMPORTANCE: Determining the relevant amino acids involved in antigenic drift on the surface protein hemagglutinin (HA) is critical to understand influenza virus evolution and efficient assessment of vaccine strains relative to current circulating strains. We used antigenic cartography to generate an antigenic map of the H9 hemagglutinin (HA) using sera produced in one of the most relevant minor poultry species, Japanese quail. Key antigenic positions were identified and tested to confirm their impact on the antigenic profile. This work provides a better understanding of the antigenic diversity of the H9 HA as it relates to reactivity to quail sera and will facilitate a rational approach for selecting more efficacious vaccines against poultry-origin H9 influenza viruses in minor poultry species.

Expression of mRNAs Encoding Hypothalamic Small Proteins, Neurosecretory Protein GL and Neurosecretory Protein GM, in the Japanese Quail, Coturnix japonica.

Neurosecretory protein GL (NPGL) and neurosecretory protein GM (NPGM) are novel neuropeptides that have been discovered in the hypothalamic infundibulum of chickens. NPGL and NPGM play important roles in lipid metabolism in juvenile chickens. The physiological functions of NPGL and NPGM in sexually mature birds remain unknown. The Japanese quail (Coturnix japonica) seems to be an appropriate model for analyzing NPGL and NPGM during sexual maturity. However, studies on NPGL or NPGM have yet to be reported in the Japanese quail. In the present study, we identified cDNAs encoding precursor proteins of NPGL and NPGM in the quail hypothalamus. In situ hybridization revealed that NPGL mRNA-expressing cells in the hypothalamus were localized in the infundibular nucleus and median eminence, and NPGM mRNA-expressing cells were only found in the mammillary nucleus. Immunohistochemistry revealed that NPGM-like immunoreactive cells were distributed in the mammillary nucleus, whereas NPGL-like immunoreactive cells were not detected in the hypothalamus. Real-time PCR analysis indicated that the expression of NPGL mRNA was higher in the hypothalamus of females than in that of males, and NPGM mRNA expression showed no sex differences. NPGL and NPGM mRNA expression in males was upregulated after 24 h of food deprivation. In females, only NPGM mRNA expression was increased by fasting. These results suggest that the physiological functions of NPGL and NPGM are different in quail, and these factors are involved in sex differences in energy metabolism.

In ovo o,p'-DDT exposure induces malformation of reproductive organs and alters the expression of genes controlling sexual differentiation in Japanese quail embryo.

In ovo exposure to o,p'-dichloro-diphenyl-trichloroethane (o,p'-DDT) impairs reproduction by inducing malformation of the reproductive organs in birds, although the mechanism remains unclear. Here, we examined the effects of o,p'-DDT on the development of the reproductive organs, the expression of genes controlling sexual differentiation, and the plasma concentrations of testosterone and estradiol in Japanese quail embryos. o,p'-DDT-containing sesame oil was injected into the yolk sac on Embryonic Day (E) 3 at a dose of 500, 2,000, or 8,000 µg per egg. On E15, the reproductive organs were observed; the gonads and Müllerian ducts (MDs) were sampled to measure the mRNA of steroidogenic enzymes, sex steroid receptors, anti-Müllerian hormone (AMH), and AMH receptor 2 (AMHR2); blood samples were collected to assay plasma testosterone and estradiol levels; and the gonads were used for histological analysis. o,p'-DDT dose-dependently increased the prevalence of hypertrophic MDs in females and residual MDs in males. In female MDs, o,p'-DDT dose-dependently decreased estrogen receptor (ER) α, ERß, and AMHR2 mRNA expression. o,p'-DDT dose-dependently induced left-biased asymmetry of testis size, and ovary-like tissue was found in the left testis after exposure to 8,000 µg per egg o,p'-DDT, although asymmetric gene expression did not occur. o,p'-DDT did not affect ovarian tissue but did decrease 17α-hydroxylase/C17-20 lyase mRNA expression and dose-dependently increased ERß mRNA expression. o,p'-DDT decreased plasma testosterone concentrations in females. These findings suggest that o,p'-DDT induces hypertrophy of the MDs and ovarian tissue formation in the left testis. Abnormal MD development may be linked to altered gene expression for sensing estrogens and AMH signals.

The detoxification of cadmium in Japanese quail by pomegranate peel powder.

Environmental pollution and exposure to toxic metals such as cadmium (Cd) can cause severe and chronic diseases and have significant side effects on vital organs. The present study aimed to evaluate the effect of pomegranate peel on biochemical factors and lipid peroxidation in intoxication by Cd in Japanese quail. Two hundred seventy quails in different groups were fed diets containing Cd and pomegranate peel from 6 to 35 days old. Then, serum biochemical parameters were assessed, including liver enzymes, urea, and thiobarbituric acid. In the quails, Cd significantly increased MDA, urea, and AST (P < 0.05). Adding pomegranate peel at 1.5 and 2% levels decreased these parameters significantly (P < 0.05). In conclusion, dietary enrichment using pomegranate peel reduced the adverse effects of Cd by improving lipid peroxidation, aspartate aminotransferase (AST), and urea in Japanese quail.

Some productive and reproductive performance, eggshell quality, serum metabolites and immune responses due to L-threonine supplementation in Japanese quail breeders' diet.

A dose-response experiment was conducted to evaluate the effects of graded levels of dietary digestible threonine (dThr) during the first laying cycle on productive and reproductive performance, egg quality and immune responses of Japanese quail breeders (Coturnix coturnix japonica). Also, dThr requirements were determined based on nutrient dose-response data. A total of 450 (360 females and 90 males) 11-week-old breeders were allocated to five increment (+0.03%) levels of dThr (0.49%, 0.52%, 0.55%, 0.58%, 0.61% and 0.64%) with five replicates per treatment and 15 (12 females and three males) birds each. The experiment lasted for 12 weeks. In response to increasing dietary dThr levels, egg production, egg mass, feed efficiency, egg specific gravity, eggshell relative weight, eggshell thickness, egg fertility (EF) and immune response against sheep red blood cell (SRBC) inoculation were improved with quadratic trends and egg hatchability as set eggs was improved with linear trends. Japanese quail breeders fed a diet with 0.58% dThr concentration (threonine/lysine ratio of 59%) showed the productive performance traits, EF, eggshell quality and immune response against SRBC inoculation in the highest values. However, feed intake, egg weight, egg albumen and yolk relative weight, egg shape index, haugh unit and egg composition were not affected by increasing dietary dThr level. Based on the broken-line regression model, the dThr requirements to optimize productive performance, eggshell quality, EF and immune response against SRBC inoculation were estimated at 159-188, 169-183, 175 and 178 mg/bird per day, respectively. It is concluded, in the Japanese quail breeders during the first laying phase a daily dThr intake of 188 mg/bird, dietary dThr concentration at 0.58% (threonine/lysine ratio of 59%) is adequate for optimized productive and reproductive performance, eggshell quality and immune responses. The estimated requirements depend on what production parameter is taken into considered for optimization.

Contribution of birds to the study of sexual differentiation of brain and behavior.

Behavioral neuroendocrinology has largely relied on mammalian models to understand the relationship between hormones and behavior, even if this discipline has historically used a larger diversity of species than other fields. Recent advances revealed the potential of avian models in elucidating the neuroendocrine bases of behavior. This paper provides a review focused mainly on the contributions of our laboratory to the study of sexual differentiation in Japanese quail and songbirds. Quail studies have firmly established the role of embryonic estrogens in the sexual differentiation of male copulatory behavior. While most sexually differentiated features identified in brain structure and physiology result from the different endocrine milieu of adults, a few characteristics are organized by embryonic estrogens. Among them, a sex difference was identified in the number and morphology of microglia which is not associated with sex differences in the concentration/expression of neuroinflammatory molecules. The behavioral role of microglia and neuroinflammatory processes requires further investigations. Sexual differentiation of singing in zebra finches is not mediated by the same endocrine mechanisms as male copulatory behavior and "direct" genetic effect, i.e., not mediated by gonadal steroids have been identified. Epigenetic contributions have also been considered. Finally sex differences in specific aspects of singing behavior have been identified in canaries after treatment of adults with exogenous testosterone suggesting that these aspects of song are differentiated during ontogeny. Integration of quail and songbirds as alternative models has thus expanded understanding of the interplay between hormones and behavior in the control of sexual differentiation.

Food patch use of Japanese quail (Coturnix japonica) varies with personality traits.

BACKGROUND: The classic optimal foraging theory (OFT) predicts animals' food patch use assuming that individuals in a population use the same strategy while foraging. However, due to the existence of animal personality, i.e. repeatable inter-individual differences and intra-individual consistency in behaviours over time and/or across contexts, individuals often exhibit different behavioural strategies, challenging the basic assumptions of the OFT. Here, we tested whether personality traits (boldness and exploration in open arena) of Japanese quail (Coturnix japonica, 38 females and 34 males) influenced their patch use in two foraging experiments with different inter-patch distances (i.e. 2 m in Experiment 1 and 3 m in Experiment 2). RESULTS: The total feeding time and food intake of individuals did not differ between Experiment 1 and 2, but in both experiments, proactive (i.e. bolder and more explorative) individuals had longer feeding time and higher food intake than reactive individuals. In Experiment 1, proactive quails changed patches more frequently and had shorter mean patch residence time than reactive individuals, while the effects were not significant in Experiment 2. The quails reduced patch residence time along with feeding, and this trend was weakened in Experiment 2 which had longer inter-patch distance. CONCLUSIONS: The above results suggest that personality traits affect animals' patch use, while the effects might be weakened with longer inter-patch distance. Our study highlights that animal personality should be considered when investigating animals' foraging behaviours because individuals may not adopt the same strategy as previously assumed. Furthermore, the interaction between personality traits and inter-patch distances, which is related to movement cost and capacity of information gathering, should also be considered.

Withania somnifera inhibits photorefractoriness which triggers neuronal apoptosis in both pre-optic and paraventricular hypothalamic area of Coturnix coturnix japonica: involvement of oxidative stress induced p53 dependent Caspase-3 mediated low immunoreactivity of estrogen receptor alpha.

Light has a very important function in the regulation of the normal physiology including the neuroendocrine system, biological rhythms, cognitive behavior, etc. The variation in photoperiod acts as a stressor due to imbalance in endogenous hormones. Estrogen and its receptors ER alpha and beta play a vital role in the control of stress response in birds. The study investigates the estrogenic effects of a well-known medicinal plant Withania somnifera (WS), mediated by estrogen receptor alpha (ERα) in the hypothalamic pre-optic area (POA) and paraventricular nuclei (PVN). Further the study elucidates its anti-oxidants and anti-apoptotic activities in the brain of Japanese quail. To validate this hypothesis, mature male quails were exposed to long day length for 3 months and then transferred to intermediate day length to become photorefractory (PR) while controls were still continued under long daylength. Supplementation of WS root extract in PR quail increases plasma estrogen and lowers corticosterone. Further, in PR quail the variation in light downregulates immunoreactivity of ERα, oxidative stress and antioxidant enzyme activities i.e. superoxide dismutase and catalase in the brain. Neuronal apoptosis was observed in the POA and PVN of PR quail as indicated by the abundant expression of Caspase-3 and p53 which reduces after the administration of WS root extract. The neuronal population also found to decrease in PR although it increased in WS administered quails. Further, the study concluded that change in photoperiod from 3 months exposure of 16L: 8D to 13.5L: 10.5D directly activates neuronal apoptosis via expression of Caspase3 and p53 expression in the brain and increases neuronal and gonadal oxidative stress while WS root extract reverses them via enhanced estrogen and its receptor ERα expression in the hypothalamic pre-optic and PVN area of Japanese quail.

Elucidating the central anorexigenic mechanism of glucagon-like peptide 1 in Japanese quail (Coturnix japonica).

Glucagon-like peptide 1 (GLP-1) elicits a potent reduction in food intake, although the central mechanism mediating this appetite-suppressive effect is not fully understood in all species. To begin to elucidate the molecular mechanisms in quail, we administered GLP-1 via intracerebroventricular (ICV) injection to 7-day-old Japanese quail (Coturnix japonica) and determined effects on food and water intake, behavior, and brain nucleus activation. We observed a reduction in food and water intake, with the lowest effective dose being 0.01 nmol. Quail injected with GLP-1 displayed fewer steps, feeding pecks, exploratory pecks, and jumps, while time spent sitting increased. We quantified c-Fos immunoreactivity at 60 min post-injection in hypothalamic and brainstem nuclei that mediate food intake and determined that the hypothalamic paraventricular nucleus (PVN), and nucleus of the solitary tract and area postrema of the brainstem were activated in response to GLP-1. In conclusion, these results suggest that GLP-1 induces anorexigenic effects that are likely mediated at the level of the PVN and brainstem.

Genetic evaluation for survival traits in Japanese quail line selected for fast growth rate.

The main objectives of this study were to estimate genetic parameters for the survival, longevity and evaluate risk factors for the occurrence of mortality in a Japanese quail line selected for high growth rate during the period from hatch to 21 days of age (GR1-21) for eight generations and its control. Total number of 1095, 2289 and 16,506 for sires, dams and progeny, respectively, was used to estimate genetic parameters, a separate hatch of 687 chicks was used to examine the risk factors of quails in the selected (SL, 438) and control (CL, 249) lines. The proportion of censored quails until 42 days of age was 82.20 and 87.14 for SL and CL, respectively. The CL showed higher longevity than SL (38.42 vs. 36.86 days). In the two tested lines, mortality% significantly declined when body weight at death increased, however, the CL had a higher reduction of mortality% than the SL (50 vs. 42%). Survival and longevity had low heritability values, low genetic and phenotypic correlations between survival and longevity with GR1-21 and ranging from 0.025 to 0.208. The survival tended to be less correlated with GR1-21 and body weight at marketing age than the longevity.

Effect of magnetized water on blood indices and histomorphometric parameters of intestinal sections of Japanese quail.

A two-way experimental design was used to demonstrate the physiological effects of magnetized water and sex on blood indices and histomorphometric parameters of Japanese quail intestine sections. Red blood cell count (RBCs), packed cell volume (PCV), hemoglobin concentration (Hb), thrombocytes, white blood cell count (WBCs), and WBC differentiation were investigated. A total of 450 unsexed Japanese quail were randomized into three groups (45/replicate; 3 replicates; 135/group). As a monitoring group, the first group was given untreated tap water to drink. The two others were consumed magnetized water that were subjected to an electrical magnetic field with a power of 1 Tesla (10,000 Gauss) and 2 Tesla (20,000 Gauss), respectively. The treatments had a significant (p ≤ 0.05) effect on thrombocytes and Hb. Sex showed significant (p ≤ 0.05) differences for RBCs and PCV at 42 days of age. At different ages, significant effects were observed on histomorphometric parameters of the Japanese quail intestinal tract. It may be inferred that the influence of magnetized water, up to 1 Tesla, was positive on the haematological and histomorphometric parameters of the Japanese quail intestinal tract by augmenting the haematological measurements, which were within a normal range and increasing the surface area of the villus.

Impact of spraying eggs with betaine after exposure to short-term thermal stress during early embryogenesis on pre and post-hatch performance of Japanese quail.

It is essential to understand and manage environmental factors for good quail production and welfare. One of the most important environmental stressors that hinder quail productivity is heat stress. This study aimed to evaluate the impact of spraying Japanese quail (Coturnix coturnix japonica) eggs with betaine after exposure to short-term high temperature during early embryogenesis on pre and post-hatch performance of quail. A total of 750 eggs were equally divided into two groups. Eggs in the first group were incubated at normal incubation temperature (37.5 °C/NIT), while those in the second group were incubated at high incubation temperature (39.0 °C/HIT) for 3 h daily from day 4-6 of incubation. Eggs in both groups were subjected to five treatments, NC (negative control), PC sprayed distilled water (positive control), while B0.5, B1, and B2 treatments were sprayed with distilled water supplemented with 500, 1000, and 2000 mg betaine/L, respectively. The chick weight at hatch, slaughter weight, and first egg weight was significantly impaired by the HIT treatment. The HIT group revealed a significant increase in cloacal temperature, H/L ratio, liver enzymes, triglyceride, and cholesterol and a significant decrease in hatchability, T3 hormone, and blood protein levels than the NIT group. Regarding betaine effects, the embryonic mortality rates, hatchability, hatched chick weight, and oviduct percentage in groups treated with 1000 or 2000 mg betaine/L were significantly improved compared with the control. Also, spraying betaine at 1000 or 2000 mg/L significantly increased blood protein and triiodothyronine (T3) hormone levels and significantly decrease liver enzyme levels and total feed consumption compared with the untreated group. The right/total ventricle ratio (RV/TV) of quail in HIT group was significantly increased, while betaine treatment significantly decreased this ratio. Considering these results, it is strongly suggested that spraying of betaine on eggs at 2000 mg/L optimizes Japanese quail performance.

Dietary tea tree (Melaleuca alternifolia) essential oil as alternative to antibiotics alleviates experimentally induced Eimeria tenella challenge in Japanese quails.

Coccidiosis is one of the most common infectious diseases seen in Japanese quails. The current study was conducted to evaluate the impact of tea tree essential oil (TTEO) on growth performance and intestinal health of quails in response to Eimeria tenella challenge. A total of 250 Japanese quails were divided into five treatments: untreated uninfected (negative control); untreated infected (positive control); infected + Amprolium; infected and 1% TTEO; infected and 2% TTEO. Except negative control, all groups were orally dosed with 5 × 104 sporulated oocysts of E. tenella. The results revealed that supplementation of 1% TTEO and treatment of amprolium improved feed intake, weight gain and feed conversion ratio in infected quails compared to the positive control. Similarly, lesion score and mortality was significantly (p < 0.01) reduced in quails supplemented with 2% TTEO and amprolium treated birds. Moreover, oocysts counts and histological features of caecum in infected birds were reversed in 1% TTEO and amprolium treatment. The histological findings of amprolium and 1% TTEO supplemented quails showed intact intestinal villi with mild sloughed epithelium. In conclusion, 1% TTEO can be safely used to control coccidiosis in Japanese quails as natural effective compound.

Ameliorative impacts of sodium humate on heat-stressed laying Japanese quail (Coturnix coturnix Japonica).

A total of 300 laying Japanese quails (230.10 ± 20 g body weight) divided into four groups (15 birds in 5 replicates/group) were used to examine the impacts of dietary sodium humate (SH) supplementation at levels of 0% (control diet), 0.2%, 0.4% and 0.6% on egg variables and physiological merits of laying quails for 10 weeks under heat stress conditions (15 June and 23 August 2021). Results showed 0.4% SH increased (p < 0.05) weight (12.27 vs. 11.91 g), production (79.84% vs. 69.20%), mass (597.13 vs. 510.48 g) and brokenness (2.8% vs. 5.4%) of eggs as compared to control. Egg shape, shell thickness, shell strength and cholesterol content as well as feed conversion ratio were higher (80.2, 295.8 µm, 1.468 kg/cm,2 11.08 mg/g and 2.69, p < 0.05) in 0.4% SH than in control group (75.2, 279.0 µm, 1.304 kg/cm,2  14.94 mg/g and 2.76). Feed intake, percentages of eggs' shells, yolk, albumen and serum biochemistry (total protein, albumin, AST and HDL) were not altered with the dietary SH. Birds fed on SH diets showed higher levels of globulin, calcium and phosphorus, as well as lower contents of albumin/globulin ratio, triglycerides, cholesterol, corticosterone compared with the control. Regression analysis of antioxidants expected higher total antioxidant capacity (TAC), superoxide dismutase, glutathione peroxidase at 0.35%, and glutathione at 0.40% SH, while the lowest concentration of malondialdehyde was computed at 0.45%. Similarly, immunoglobulins (IgG and IgM) maximum values were determined at 0.35% and 0.40% levels. Moreover, the concentration of tumour necrosis factor-alpha increased (p < 0.05) in all SH levels as compared to the control group. It is conceivable to conclude that the dietary implementation of SH at a level of 0.4% improved egg variables and well-being aspects of laying quail exposed to heat stress conditions.

Impact of different light intensities to Japanese quail eggs on hatching results and post-hatch performance.

The study aims to evaluate dichromatic light with different intensities during incubation on pre- and post-hatch performance of Japanese quail. In completely randomized design, 600 quail hatching eggs were evenly divided into 4 treatments, 3 replicates, and 50 eggs in each. These eggs were provided with lighted incubation [dichromatic light; green-red (GR) with three different intensities 150, 250, and 350 lx] with lighting schedule (12L: 12D), and one treatment was the complete dark period during incubation and considered as control group. After hatch, 300 quail chicks were equally divided into four treatments (3 replicates and 25 chicks in each). Effects of GR light with different intensities during incubation were evaluated on hatching traits and post-hatch performance of Japanese quails. Regarding hatching traits, hatchability was better in all the treated groups (150, 250, and 350 lx of GR light during incubation) compared to complete dark. In terms of embryonic mortality, early embryonic mortality was lower in the 250 lx group, mid embryonic mortality in dark, and late embryonic mortality in the 150 lx group. Furthermore, an ideal chick spread and growth performance (weight gain, feed intake, feed conversion ratio, and liveability) was reported in 250 lx group compared to other treated groups. Serum chemistry values were higher in dark group compared to 150, 250, and 350 lx groups. Pre-slaughter weight was higher in 250 and 350 lx group than 150 lx and dark group, whereas carcass weight was better in 350 lx group compared to dark. In conclusion, stimulation of dichromatic light (green + red) at 250 lx to Japanese quail eggs positively influenced hatching traits and post-hatch growth performance of Japanese quail.

The comparison of two different plumage-color lines of Japanese quail (Coturnix japonica) disclosed a significant effect in increasing abdominal fat contents with increasing age.

This study aimed to investigate the characteristic differences between the white and brown-feathered Japanese quails, by evaluating the carcass traits and egg fat content, blood parameters, and intestinal histopathological features. A total of 1200 1-day-old Japanese quail chicks of two varieties (brown and white-feathered) were used in this study. Live body weight and feed intake were reordered every week. At the 4th week of age, 80 birds from each variety were slaughtered and carcass quality measurements and histopathological changes were recorded. After 6 weeks of age, eggs were collected, and egg quality was assessed. The results revealed that white-feathered quails had significantly heavier body weights and higher growth rates. At 4 weeks of age, females of the white-feather quail had significantly heavier slaughter, after de-feathering, and carcass weights. Remarkable variations between the studied quail varieties, with significant dominance of females in both varieties, at the level of water holding capacity, pH, and meat tenderness ascertained an obvious superiority of white-feathered quails compared to brown ones and indicated the higher tendency of the white quails for meat production. These results were linked with significant changes in biochemical profiles including lipids biomarkers, total protein, and Ca and phosphorus levels along with variations in the intestinal morphometry. It can be concluded that white-feathered quails had, in general, higher values of productivity compared with the brown-feathered ones during growing and laying periods.

Residual feed intake breeding value associated with growth, carcass traits, meat quality, bone properties and humoral immunity in Japanese quail.

Improved feed efficiency (FE) is one of the most important achievements in animal breeding programs. The present investigation aimed to determine the phenotypic correlations of residual feed intake (RFI) breeding value with some traits in growing Japanese quail. A total of 48 birds (24 males and 24 females) were selected from 220 quail chicks divided into three classes based on breeding values (BV) for RFI including low (LBV; n = 16), medium (MBV; n = 16), and high-BV (HBV; n = 16) were used to test FE. The effects of three groups of BV on the studied traits including carcass traits, meat quality, humoral immunity, and bone properties were evaluated. The BV for RFI was positively correlated with feed conversion ratio (FCR) and feed intake (FI) but not with metabolic BW (MBW0.75). Live body weight, carcass, breast, and thigh weight in the LBV-RFI group were significantly greater than those in the HBV-RFI group. The BV for RFI had a negative correlation with live body weight and thigh weight. Our findings suggested that the selection of LBV-RFI quails may be useful to increase live body weight without any adverse impact on meat quality and bone properties, and live body weight can be implemented in breeding programs as an indirect selection indicator for improvement of FE in quails.

Effects of prenatal dichromatic light exposure on hatching results and post-hatch performance of Japanese quail.

Present study evaluated the effect of lighted incubation on pre- and post-hatch performance of Japanese quail. In a completely randomized design, 1200 eggs were evenly divided into 4 treatments groups having six replicates (each tray was considered as replicate), 50 eggs each. Different dichromatic lights (Green + Red; GR, Green + Blue; GB, and Blue + Red; BR) of 250 lux were provided during incubation for 12 h daily and effects of these lights very evaluated on hatching results and post-hatch growth. After hatch, 600 quail chicks were divided into 4 treatments, 6 replicates, and 25 birds each. Regarding hatching traits, better hatchability was found in the GR group compared to GB, BR, and dark group; while early embryonic mortality was lower in BR, GB, and dark group than GR; mid embryonic mortality was lower in dark group and late embryonic mortality was noted in the GR group than those of other treatment groups. In addition, moisture loss during incubation was minimum in BR and dark groups; however, chick spread was better in the GR group. In terms of growth performance, weight gain was better in the GR group; feed intake in dark, feed conversion ratio in BR, and livability were better in BR and GR group. In morphometrics, keel and shank length were higher in all the colored groups (GB, BR, and GR) whereas body length, wing spread, shank circumference, drumstick length, and circumference were higher in the GR group. Regarding serum chemistry, glucose, albumin, and globulin levels were higher in the GR group. It was concluded that under the experimental conditions, GR light at the prenatal stage to Japanese quail eggs positively influenced hatching performance and post-hatch growth.

Assessment of variations in productive performance of two different plumage color varieties of Japanese quail and their reciprocal crosses.

This study aimed to detect the phenotypic differences between the brown (BB) and white (WW) feathered quails and their reciprocal crosses (BW and WB) over two successive generations. The WW and cross quails, especially the BW, had the heaviest body weights, throughout the studied period, with significant variations between the two studied generations (P<0.05). Moreover, the WW and BW possessed the largest egg production during the F1, while in the F2, the BB had superiority among the studied quails with a prominent superiority of the F2 over the F1 (P<0.05). However, the F1 had higher egg weights than F2 with superiority of WW quails compared to the others (P<0.05). Also, the WW quails had the lowest lipid contents of the eggs. These phenotypic variations among the studied quails might be preliminarily explained by the results of the analyzed microsatellite markers despite the few markers used. The high variability among the BW and WB quails might be due to the larger number of alleles (NA and Ne) and the lower values of FIS with low heterozygosity levels (HO and He). Moreover, the BW and BB were the closest, while WB and WW were the farthest because of the high and low genetic identities and the high and low genetic distance between them, respectively. So the obtained results might introduce an initial scientific basis for evaluating and employing the genetic properties of BB, WW, BW, and WB quails in further genetic improvement program, and more microsatellite markers are recommended.

Cyclin D1 gene expression is essential for cell cycle progression from the maternal-to-zygotic transition during blastoderm development in Japanese quail.

Embryogenesis proceeds by a highly regulated series of events. In animals, maternal factors that accumulate in the egg cytoplasm control cell cycle progression at the initial stage of cleavage. However, cell cycle regulation is switched to a system governed by the activated nuclear genome at a specific stage of development, referred to as maternal-to-zygotic transition (MZT). Detailed molecular analyses have been performed on maternal factors and activated zygotic genes in MZT in mammals, fishes and chicken; however, the underlying mechanisms remain unclear in quail. In the present study, we demonstrated that MZT occurred at blastoderm stage V in the Japanese quail using novel gene targeting technology in which the CRISPR/Cas9 and intracytoplasmic sperm injection (ICSI) systems were combined. At blastoderm stage V, we found that maternal retinoblastoma 1 (RB1) protein expression was down-regulated, whereas the gene expression of cyclin D1 (CCND1) was initiated. When a microinjection of sgRNA containing CCND1-targeted sequencing and Cas9 mRNA was administered at the pronuclear stage, blastoderm development stopped at stage V and the down-regulation of RB1 did not occur. This result indicates the most notable difference from mammals in which CCND-knockout embryos are capable of developing beyond MZT. We also showed that CCND1 induced the phosphorylation of the serine/threonine residues of the RB1 protein, which resulted in the degradation of this protein. These results suggest that CCND1 is one of the key factors for RB1 protein degradation at MZT, and the elimination of RB1 may contribute to cell cycle progression after MZT during blastoderm development in the Japanese quail. Our novel technology, which combined the CRISPR/Cas9 system and ICSI, has the potential to become a powerful tool for avian-targeted mutagenesis.