Signaling pathways and regulatory networks in quail skeletal muscle development: insights from whole transcriptome sequencing.

Quail, as an advantageous avian model organism due to its compact size and short reproductive cycle, holds substantial potential for enhancing our understanding of skeletal muscle development. The quantity of skeletal muscle represents a vital economic trait in poultry production. Unraveling the molecular mechanisms governing quail skeletal muscle development is of paramount importance for optimizing meat and egg yield through selective breeding programs. However, a comprehensive characterization of the regulatory dynamics and molecular control underpinning quail skeletal muscle development remains elusive. In this study, through the application of HE staining on quail leg muscle sections, coupled with preceding fluorescence quantification PCR of markers indicative of skeletal muscle differentiation, we have delineated embryonic day 9 (E9) and embryonic day 14 (E14) as the start and ending points, respectively, of quail skeletal muscle differentiation. Then, we employed whole transcriptome sequencing to investigate the temporal expression profiles of leg muscles in quail embryos at the initiation of differentiation (E9) and upon completion of differentiation (E14). Our analysis revealed the expression patterns of 12,012 genes, 625 lncRNAs, 14,457 circRNAs, and 969 miRNAs in quail skeletal muscle samples. Differential expression analysis between the E14 and E9 groups uncovered 3,479 differentially expressed mRNAs, 124 lncRNAs, 292 circRNAs, and 154 miRNAs. Furthermore, enrichment analysis highlighted the heightened activity of signaling pathways related to skeletal muscle metabolism and intermuscular fat formation, such as the ECM-receptor interaction, focal adhesion, and PPAR signaling pathway during E14 skeletal muscle development. Conversely, the E9 stage exhibited a prevalence of pathways associated with myoblast proliferation, exemplified by cell cycle processes. Additionally, we constructed regulatory networks encompassing lncRNA‒mRNA, miRNA‒mRNA, lncRNA‒miRNA-mRNA, and circRNA-miRNA‒mRNA interactions, thus shedding light on their putative roles within quail skeletal muscle. Collectively, our findings illuminate the gene and non-coding RNA expression characteristics during quail skeletal muscle development, serving as a foundation for future investigations into the regulatory mechanisms governing non-coding RNA and quail skeletal muscle development in poultry production.

Glucose oxidation drives trunk neural crest cell development and fate.

Bioenergetic metabolism is a key regulator of cellular function and signaling, but how it can instruct the behavior of cells and their fate during embryonic development remains largely unknown. Here, we investigated the role of glucose metabolism in the development of avian trunk neural crest cells (NCCs), a migratory stem cell population of the vertebrate embryo. We uncovered that trunk NCCs display glucose oxidation as a prominent metabolic phenotype, in contrast to what is seen for cranial NCCs, which instead rely on aerobic glycolysis. In addition, only one pathway downstream of glucose uptake is not sufficient for trunk NCC development. Indeed, glycolysis, mitochondrial respiration and the pentose phosphate pathway are all mobilized and integrated for the coordinated execution of diverse cellular programs, epithelial-to-mesenchymal transition, adhesion, locomotion, proliferation and differentiation, through regulation of specific gene expression. In the absence of glucose, the OXPHOS pathway fueled by pyruvate failed to promote trunk NCC adaptation to environmental stiffness, stemness maintenance and fate-decision making. These findings highlight the need for trunk NCCs to make the most of the glucose pathway potential to meet the high metabolic demands appropriate for their development.

Do thermal acclimation and an acute heat challenge alter myonuclear domain of control- and fast-growing quail?

Efforts to determine physiological traits that may render species resilient or susceptible to changing global temperatures have accelerated in recent years. Temperature is of critical importance to biological function; thus, climate change has the potential to severely affect all levels of biological organization in many species. For example, increases in environmental temperatures may alter muscle structure and function in birds. Myonuclear domain (MND), an under-studied aspect of avian muscle physiology that changes in response to thermal stress, is defined as the amount of cytoplasm within a muscle fiber that each nucleus is responsible for servicing. Here, we used two random bred lines of Japanese quail (Coturnix japonica) representing examples of control and fast growth rates. We used a factorial design to administer four treatment combinations to each line - an initial period of either heat-stress acclimation (Acclimation) or no acclimation (Not acclimated) followed by either a heat-stress challenge (HS) or no challenge (NC) after week 8 of age - to determine the effects of thermal acclimation and acute thermal stress on quail MND. We found a significant interaction between line * final treatment with fast-growing, HS birds demonstrating the lowest numbers of nuclei per mm of fiber, and Acclimated control-growing birds showing the highest numbers of nuclei per mm of fiber. There was a significant effect of line on MND with the fast-growing line having larger MNDs. Initial treatment with Not Acclimated birds showed larger MNDs. Additionally, control growing quail demonstrated positive correlations with fiber size, whereas fast growing quail did not. This may mean that nuclei in larger fibers of fast-growing quail may be functioning maximally, and that increases in temperature may also demonstrate similar effects.

Impact of acute short-term high thermal stress during early embryogenesis on hatchability, physiological body reaction, and ovarian follicles development of quails.

This experiment aimed to evaluate the impact of continuous and intermittent thermal stress during early embryogenesis on hatchability, physiological body reaction, ovary weight, and follicle development of quails. A total of 540 eggs were divided into 3 equal groups (3 groups × 6 replicates × 30 eggs). In the first group (control), eggs were incubated at normal incubation conditions (37.5°C and 50-55% relative humidity) from day 0 till hatching. In the second group (continuous thermal stress [CTS]), eggs were daily exposed to 39.5°C and 50 to 55% during the early embryogenesis for 3 successive days (E4-E6) for 3 h (12:00-15:00). In the third group (intermittent thermal stress [ITS]), eggs were daily exposed to 39.5°C and 50 to 55% during the early embryogenesis for 90 min (12:00-13:30) then temperature was returned to 37.5°C for 60 min (13:30-14:30) after that the temperature was raised again for 39.5°C for 90 min (14:30-16:00) daily for 3 successive days (E4-E6). The findings showed that the highest relative water loss form egg (RWL/%) at 6 d of incubation was obtained in the CTS group (P ≤ 0.05). The hatchability rate was significantly (P ≤ 0.05) decreased in the thermal-treated groups compared with the control group. The body surface temperature and cloacal temperature in the CTS and ITS groups significantly (P ≤ 0.001) increased compared with the control group. Chick weight (g) at 5 wk old, total weight gain, daily weight gain were significantly lower (P ≤ 0.05) in the CTS group compared with the control group. Triiodothyronine (T3) hormone concentration and globulin level were significantly (P ≤ 0.05) lower in the CTS and ITS groups compared with the control. The ovarian follicle weights (first, second, third, fourth, and fifth) and the diameter of the large follicle (fifth follicle) were significantly (P ≤ 0.01) decreased by increasing incubation temperature. From these findings, it could be concluded that the hatchability and body weight at sexual maturity for quails produced from eggs exposed to CTS and IST were significantly decreased by 8 and 2.1% as well as 2.98 and 2.1%, respectively, compared with the control group.

Light at night reduces digestive efficiency of developing birds: an experiment with king quail.

Artificial light at night (ALAN) exposes animals to a novel environmental stimulus, one that is generally thought to be maladaptive. ALAN-related health problems have received little attention in non-model species, and we generally know little about the nutritional-physiological impacts of ALAN, especially in young animals. Here, we use a novel application of the acid steatocrit method to experimentally assess changes in digestive efficiency of growing king quail (Excalfactoria chinensis) in response to ALAN. Two weeks after hatching, quail were split into two groups (n = 20-21 per group): overnight-light-treated vs. overnight-dark-treated. When the chicks were 3 weeks old, the experimental group was exposed to weak blue light (ca. 0.3 lux) throughout the entire night for 6 consecutive weeks, until all the chicks had achieved sexual maturation. Fecal samples for assessing digestive efficiency were collected every week. We found that digestive efficiency of quail was reduced by ALAN at two time points from weeks 4 to 9 after hatching (quail reach adulthood by week 9). The negative effect of ALAN on digestion coincided with the period of fastest skeletal growth, which suggests that ALAN may reduce digestive efficiency when energetic demands of growth are at their highest. Interestingly, growth rate was not influenced by ALAN. This suggests that either the negative physiological impacts of ALAN may be concealed when food is provided ad libitum, the observed changes in digestive efficiency were too small to affect growth or condition, or that ALAN-exposed birds had reduced energy expenditure. Our results illustrate that the health impacts of ALAN on wild animals should not be restricted to traditional markers like body mass or growth rate, but instead on a wide array of integrated physiological traits.

Growth performance, hemato-biochemical indices, thyroid activity, antioxidant status, and immune response of growing Japanese quail fed diet with full-fat canola seeds.

Dietary supplementation of full-fat canola seeds (FFCS) can be part of the global solution to the problem of incessant raise in feed cost. Genetic improvement via plant breeding constantly introduces varieties of canola with higher content of unsaturated fatty acids and lower content of anti-nutritional factors. This experiment was conducted to evaluate the dietary inclusion effect of new mutant of FFCS on growth performance, carcass traits, blood metabolites, thyroid activity, antioxidant status, and immune response of growing Japanese quails. A total of 640 21-day-old quails were used in a completely randomize design with and equally allotted to four experimental groups and eight replicates each. The experimental groups fed diets supplemented with 0, 50, 100, and 150 g FFCS/kg diet from 21 to 49 days of age. All experimental diets were isocaloric and isonitrogenous. The inclusion level of FFCS did not affect body weight and body weight gain at all studied periods, while feed intake was reduced and feed conversion ratio was enhanced linearly under all experimental periods except the duration from 28 to 35 days of age. All carcass traits were not statistically affected with graded levels of FFCS. Hemoglobin, leucocytes, mean corpuscular hemoglobin (MCH), and MCH concentration values were linearly increased. Serum protein fractions, liver enzymes activities, renal function biomarkers, and triiodothyronine concentration were not affected. However, serum lipid profile was significantly altered, where high-density lipoprotein was linearly increased while triglycerides and low-density lipoprotein (LDL) were linearly (and quadratically for LDL) decreased. Antioxidant and immune statuses were improved; significant increment in glutathione and glutathione peroxidase values and antibody titer against Newcastle disease virus were elevated in groups treated with 10% and 15% FFCS. In conclusion, studied levels of FFCS, however, were not able to promote the growth rate of growing Japanese quails; they were more effective in boosting their antioxidant status and humoral immune response.

Use of red pepper oil in quail diets and its effect on performance, carcass measurements, intestinal microbiota, antioxidant indices, immunity and blood constituents.

Plant-derived additives are used to maintain the health and growth performance of livestock. The use of red pepper oil (RPO) has recently attracted considerable scientific interest mainly due to its potential benefits for animals and humans. The present study was conducted to evaluate the effect of dietary supplementation with RPO on growth performance, carcass measurements, antioxidant status and immunity of growing quails between 1 and 5 weeks of age. A total of 240 growing quails (1-week old) were distributed into 5 equal groups consisting of 48 birds (4 replicates of 12 birds each). The first group was fed a basal diet without RPO (0 g/kg diet), and the second, third, fourth and fifth groups received diets containing RPO (0.4, 0.8, 1.2, 1.6 g/kg diet, respectively). The experiment lasted for 5 weeks. At age of 5 weeks, quails were slaughtered for carcass examinations, microbiological analysis of intestine and to determine blood constituents. Data were statistically analyzed by one-way ANOVA. Quails fed with 0.8 g RPO/kg diet showed 12.14%, 14.4% and 15% improvement in live BW, body weight gain and feed conversion ratio, respectively, compared with the control group. Quails that received diets with 1.2 g RPO consumed more feed than the others during the total period (1 to 5 weeks). Plasma globulin levels were significantly decreased (P = 0.0102), but albumin/globulin ratio was significantly increased (P = 0.0009) in birds fed diets containing RPO (0.4 and 1.2 g/kg) compared with those in the control group. Activity of liver enzymes in the plasma was nonsignificantly decreased in quails supplemented with 0.8 g RPO/kg diet compared with those in the control group. Activities of antioxidant enzymes (glutathione and catalase) in the group fed on diets supplemented with RPO (0.8 g/kg) were significantly higher than those in the control group. The inclusion of RPO (0.8 g/kg diet) in quail diets improved (P < 0.05) plasma lipid profile and also decreased pH of the caecal content (P = 0.0280) compared with those in the control group. The caecal bacterial population, Salmonella spp., coliform and Escherichia coli, were lowered (P < 0.05) in the groups treated with RPO (0.8, 1.2 and 1.6 g/kg) compared with those in the control group. In conclusion, dietary supplementation of RPO (0.8 g/kg) can enhance the performance and antioxidant indices and decrease intestinal pathogens and thus improve the health status of Japanese quail.

Sperm quality and quantity evolve through different selective processes in the Phasianidae.

Sperm competition is often considered the primary selective force underlying the rapid and diversifying evolution of ejaculate traits. Yet, several recent studies have drawn attention to other forms of selection with the potential of exceeding the effects of sperm competition. Since ejaculates are complex, multivariate traits, it seems plausible that different ejaculate components vary in their responses to different selective pressures. Such information, however, is generally lacking as individual ejaculate traits tend to be studied in isolation. Here, we studied the macroevolutionary patterns of ejaculate volume, sperm number, sperm length and the proportion of viable normal sperm in response to varying levels of sperm competition, body size and the duration of female sperm storage in pheasants and allies (Phasianidae). Ejaculate volume, sperm number and sperm viability were all relatively higher in polygamous than in monogamous mating systems. However, whereas ejaculate volume additionally covaried with body size, sperm number instead increased with the female sperm-storage duration, in conjunction with a decrease in sperm length. Overall, our results revealed important details on how different forms of selection can jointly shape ejaculates as complex, composite traits.

Exogenous Expression of an Alternative Splicing Variant of Myostatin Prompts Leg Muscle Fiber Hyperplasia in Japanese Quail.

Myostatin (MSTN) negatively regulates muscle growth and development through inhibiting myoblast proliferation and differentiation. Five alternative splicing isoforms of MSTN (MSTN-A to MSTN-E) have been discovered in domestic avian species. MSTN-A has high expression in skeletal muscle and encodes the full-length peptide with anti-myogenic activity. Another isoform, MSTN-B, is also highly expressed in skeletal muscle and encodes a truncated peptide that has pro-myogenic capabilities in vitro, which include promoting the proliferation and differentiation of quail muscle precursor cells. The objective of this study was to investigate overexpression of MSTN-B in vivo by using two independent lines of transgenic Japanese quail with expression directed in the skeletal muscle. Unexpectedly, the chicken skeletal muscle alpha actin 1 (cACTA1) promoter resulted in restricted exogenous MSTN-B protein expression to certain skeletal muscles, such as the gastrocnemius and tibialis anterior, but not the pectoralis major muscle. Gastrocnemius weight as a percentage of body weight in transgenic quail was increased compared to non-transgenic quail at posthatch day 21 (D21) and posthatch D42. An increase in the size of the gastrocnemius in transgenic quail was attributed to an increase in fiber number but not fiber cross-sectional area (CSA). During embryonic development, paired box 7 (PAX7) expression was prolonged in the transgenic embryos, but other myogenic regulatory factors (MRFs) were unchanged after MSTN-B overexpression. Taken together, these data provide novel insights into the regulation of skeletal muscle development by alternative splicing mechanisms in avians.

Quill mite infestation of rock ptarmigan Lagopus muta (Aves: Phasianidae) in relation to year and host age, sex, body condition, and density.

A total of 1209 ptarmigan were examined for Mironovia lagopus, including 721 juvenile birds (ca. 3 months old) and 488 adult birds (15 months or older). A total of 88 birds or 7.3% (n = 1209, 95% cl 5.9-8.9%) were infested with M. lagopus. There was an age difference in prevalence of infection, and more adults (10.7%, n = 488, 95% cl 8.2-13.7%) than juveniles (5.0%, n = 721, 95% cl 3.6-6.8%) were infested. There was a significant age effect in the mean intensity index, and adult birds had more advanced infestations compared with juvenile birds. There were no significant changes in either the interannual prevalence of infection or the mean intensity index of infection. Of the feather types inspected, there was no age-related difference in selection of feathers, nor was there any preference of mites for any one of the inspected feather types. Body dispersants were all adult females. There was an age-related difference in mean intensities of infection of body dispersants; it was higher in adult birds. The methods used to determine presence or absence of M. lagopus were not 100% accurate especially for juvenile hosts, and this at least partly explains the difference in prevalence among age groups. There was no relationship between host body condition or host density and infection by M. lagopus.

Influences of stocking density and dietary probiotic supplementation on growing Japanese quail performance.

This study investigated the effects of stocking density and dietary supplementation of probiotic (Lactobacillus) on growth, traits of carcass, lymphoid organs and intestinal microbial count of Japanese quail. Quail chicks (one week old) were randomly allotted to 6 groups in a 2×3 factorial experiment included 2 levels of stocking density (100, and 75 cm2/bird) and 3 levels of probiotic (0.00, 0.02 and 0.04 g/kg diet). Quail reared at 100 cm2/ bird showed significantly the highest body weight and daily gain. Quail reared at 75 cm2/ bird had significantly the lower average of feed consumption and the best ratio of feed conversion. Increasing level of probiotic up to 0.04 g/kg led to a significant decrease in dressing percentage by about 2.7 % compared to the control. Interaction effect was insignificant on carcass traits studied except for liver percentage which was significantly higher for birds fed diet supplemented with probiotic at 0.04 g/kg of diet and reared at 100 cm2/bird. Rearing quail at 100 cm2/bird stocking density significantly increased bursa weight. Birds fed diet supplemented with probiotic at 0.02 g/kg of diet and reared at 100 cm2/bird showed significantly the highest (p = 0.043) spleen weight when compared with the other groups. In conclusion, the low (100 cm2/bird) stocking density was sufficient to give better performance of Japanese quail than the other group. Probiotic supplementation diminished the stressful effect of crowding on growing Japanese quail.

Effect of low- or high-CP diet with cold-pressed oil supplementation on growth, immunity and antioxidant indices of growing quail.

This study was to investigate the effect of dietary protein levels and supplementation with two cold-pressed oils on the performance, immunity and antioxidant indices of growing Japanese quails. A 3 × 4 factorial experiment, using 3 dietary crude protein (CP) levels (22%, 24% and 26%) and 4 supplemental herbal oils levels: control without additives, 250 mg marjoram (Mar), 250 mg thyme (Thy) and 125 mg Mar + 125 mg Thy per kg diet. A total of 720 unsexed, 7-day-old quail chicks were randomly distributed into twelve treatment groups (4 replicates per group of 15 chicks in each). The live body weight (LBW) on 42 days and body weight gain (BWG) from 7 to 42 days were significantly improved (p < 0.01 and 0.05) in chicks fed 24% or 26% CP compared with those of the 22% CP group. The groups fed diets supplemented with Mar or in combination with Thy oil exhibited significantly better LBW, BWG and feed conversion ratio (FCR) than the control at all periods except BWG from 21 to 42 days and FCR from 7 to 21 days were not significant. Feed intake was gradually decreased with increasing dietary CP level or adding herbal oils during the periods 21-42 and 7-42 days of age. Plasma triglycerides, cholesterol, total lipids and malondialdehyde were decreased by cold-pressed oils supplementation, but the activity of reduced glutathione and superoxide dismutase was increased (p < 0.01) compared with the control. In conclusion, quails fed a diet containing 24% or 26% CP or diets supplemented with Mar or Thy oils or both exhibited improvement in the performance, lipid profile, immunity and antioxidant capacity without any detrimental impacts on the other studied parameters.

Exposure to artificial light at night increases innate immune activity during development in a precocial bird.

Humans have greatly altered Earth's night-time photic environment via the production of artificial light at night (ALAN; e.g. street lights, car traffic, billboards, lit buildings). ALAN is a problem of growing importance because it may significantly disrupt the seasonal and daily physiological rhythms and behaviors of animals. There has been considerable interest in the impacts of ALAN on health of humans and other animals, but most of this work has centered on adults and we know comparatively little about effects on young animals. We exposed 3-week-old king quail (Excalfactoria chinensis) to a constant overnight blue-light regime for 6 weeks and assessed weekly bactericidal activity of plasma against Escherichia coli - a commonly employed metric of innate immunity in animals. We found that chronic ALAN exposure significantly increased bactericidal activity and that this elevation in immune performance manifested at different developmental time points in males and females. Whether this short-term increase in immune activity can be extended to wild animals, and whether ALAN-mediated increases in immune activity have positive or negative fitness effects, are unknown and will provide interesting avenues for future studies.

Effects of the stems and leaves of Astragalus membranaceus on growth performance, immunological parameters, antioxidant status, and intestinal bacteria of quail.

This study was designed to evaluate the potential application of the stems and leaves of Astragalus membranaceus (AMSL) in the poultry industry. Quails were divided into four groups and fed daily with an AMSL-free diet (control) or with 1%, 3%, or 5% (w/w) AMSL-incorporated diets for 35 days. The results showed that supplementing AMSL in the diet, especially at a concentration of 3%, increased daily gain and feed intake during the entire experiment (p < 0.05). The immune organ development of the thymus and bursa of Fabricius was promoted, and the immune system was enhanced by increasing the quantities of IgA and complements C3 and C4 (p < 0.05). The total antioxidant capacity and the activities of glutathione peroxidase and catalase were increased (p < 0.05). Moreover, the 3%-5% AMSL groups regulated the intestinal flora by promoting the proliferation of lactic acid bacteria and inhibiting the growth of coliform bacteria (p < 0.05). In conclusion, feeding incorporated diets with appropriate AMSL levels significantly increased growth performance, strengthened the immune system, improved antioxidative status, and regulated the intestinal microflora of quails, suggesting that AMSL has the potential to serve as a feed additive in the poultry industry.

Two-step Approach to Explore Early- and Late-stages of Organ Formation in the Avian Model: The Thymus and Parathyroid Glands Organogenesis Paradigm.

The avian embryo, as an experimental model, has been of utmost importance for seminal discoveries in developmental biology. Among several approaches, the formation of quail-chicken chimeras and the use of the chorioallantoic membrane (CAM) to sustain the development of ectopic tissues date back to the last century. Nowadays, the combination of these classical techniques with recent in vitro methodologies offers novel prospects to further explore organ formation. Here we describe a two-step approach to study early- and late-stages of organogenesis. Briefly, the embryonic region containing the presumptive territory of the organ is isolated from quail embryos and grown in vitro in an organotypic system (up to 48 h). Cultured tissues are subsequently grafted onto the CAM of a chicken embryo. After 10 days of in ovo development, fully formed organs are obtained from grafted tissues. This method also allows the modulation of signaling pathways by the regular administration of pharmacological agents and tissue genetic manipulation throughout in vitro and in ovo developmental steps. Additionally, developing tissues can be collected at any time-window to analyze their gene-expression profile (using quantitative PCR (qPCR), microarrays, etc.) and morphology (assessed with conventional histology and immunochemistry). The described experimental procedure can be used as a tool to follow organ formation outside the avian embryo, from the early stages of organogenesis to fully formed and functional organs.

Levels of supplementation of inorganic selenium and vitamin E for meat quail aged 0 to 14 and 14 to 35 days.

Two experiments were carried out to determine the levels of supplementation of inorganic selenium (Se) and vitamin E (VE) in diets of quails aged 0-14 and 14-35 days old. A completely randomized design was used in a factorial design (Se = 0.1125; 0.2250; 0.3375 and 0.4500 mg kg-1  diet-1  × VE = 10; 23; 36 and 49 IU kg-1  diet-1 ). In experiment 1, quail (n = 2,400) were aged 0-14 days and were divided into 16 treatments, with three replicates of 50 birds. In experiment 2, quail (n = 1,680) were aged 14-35 days and were divided into the same treatments, with three replicates of 35 birds. At age 0-14 days, the levels of VE did not affect performance (p > .05); however, the feed conversion (FC) was influenced by a quadratic effect (p = .0515), according to the level of Se, with a higher level estimated at 0.29 mg Se kg-1  diet-1 . At age 14-35 days, there was a linear effect with interaction (Se × VE), for FC (p = .0150) and weight gain (WG; p = .0266). FC (Se, p = .0048 and VE, p = .0019) and WG (Se, p = .0049 and VE, p = .0068) improved linearly with increasing levels of Se and VE. The feed intake (FI) decreased linearly (p = .0582) as a function of VE. The carcass yield showed a quadratic effect (p = .0056) on the levels of VE, with a higher yield estimation of 27.24 IU VE/kg of diet. It can be concluded that the optimum level of supplementation at age 0-14 days was 0.29 mg Se kg-1  diet-1 and 10 IU VE kg-1  diet-1 and at age 14-35 days, it was 0.4500 mg Se kg-1  diet-1 and 49 IU of VE kg-1  diet-1 .

Altered embryonic development in northern bobwhite quail (Colinus virginianus) induced by pre-incubation oscillatory thermal stresses mimicking global warming predictions.

Global warming is likely to alter reproductive success of ground-nesting birds that lay eggs normally left unattended for days or even weeks before actual parental incubation, especially in already warm climates. The native North American bobwhite quail (Colinus virginianus) is such a species, and pre-incubation quail eggs may experience temperatures ≥45°C. Yet, almost nothing is known about embryonic survival after such high pre-incubation temperatures. Freshly laid bobwhite quail eggs were exposed during a 12 day pre-incubation period to one of five thermal regimes: low oscillating temperatures (25-40°C, mean = 28.9°C), high oscillating temperatures (30-45°C, mean = 33.9°C), low constant temperatures (28.85°C), high constant temperatures (mean = 33.9°C), or commercially employed pre-incubation temperatures (20°C). After treatment, eggs were then incubated at a standard 37.5°C to determine subsequent effects on embryonic development rate, survival, water loss, hatching, and embryonic oxygen consumption. Both quantity of heating degree hours during pre-incubation and specific thermal regime (oscillating vs. non-oscillating) profoundly affected important aspects of embryo survival and indices of development and growth Pre-incubation quail eggs showed a remarkable tolerance to constant high temperatures (up to 45°C), surviving for 4.5±0.3 days of subsequent incubation, but high oscillating pre-incubation temperature increased embryo survival (mean survival 12.2±1.8 days) and led to more rapid development than high constant temperature (maximum 38.5°C), even though both groups experienced the same total heating degree-hours. Oxygen consumption was ~200-300 µl O2.egg.min-1 at hatching in all groups, and was not affected by pre-incubation conditions. Oscillating temperatures, which are the norm for pre-incubation quail eggs in their natural habitat, thus enhanced survival at higher temperatures. However, a 5°C increase in pre-incubation temperature, which equates to the predicted long-term increases of 5°C or more, nonetheless reduced hatching rate by approximately 50%. Thus, while pre-incubation bobwhite eggs may be resiliant to moderate oscillating temperature increases, global warming will likely severely impact wild bobwhite quail populations, especially in their strongholds in southern latitudes.

Fluorescent Quail: A Transgenic Model System for the Dynamic Study of Avian Development.

Real-time four-dimensional (4D, xyzt) imaging of cultured avian embryos is an ideal method for investigating the complex movements of cells and tissues during early morphogenesis. While methods that transiently label cells, such as electroporation, are highly useful for dynamic imaging, they can also be limiting due to the number and type of cells that can be effectively targeted. In contrast, the heritable, stable, and long-term expression of a fluorescent protein driven by the exogenous promoter of a transgene overcomes these challenges. We have used lentiviral vectors to produce several novel transgenic quail lines that express fluorescent proteins either ubiquitously or in a cell-specific manner. These lines have proven to be useful models for dynamic imaging and analysis. Here, we provide detailed protocols for generating transgenic quail with the emphasis on producing high titer lentivirus , effectively introducing it into the early embryo and efficiently screening for G1 founder birds .

Thymol feed supplementation in quail alters the percentages of nutritionally relevant egg yolk fatty acids: effects throughout incubation.

BACKGROUND: Polyunsaturated fatty acids (PUFA) are crucial components of egg yolk and particularly prone to oxidative damage, generating losses of nutrients for embryonic development and influencing the quality of eggs for human consumption. The aim of this study was to evaluate whether dietary thymol (a natural antioxidant) is related to changes in quail egg yolk total (T), triglyceride (TG) and phospholipid (PL) fatty acid composition at different stages of embryo development. Thus female Japanese quail (100 days of age) were assigned to one of two dietary treatments (12 individuals each): CON (basal diet) or THY (0.0016 mol thymol day-1 per bird). After 2 weeks of supplementation, eggs were incubated and samples were obtained at 0, 4 and 16 days of embryonic development. RESULTS: In 0 day THY eggs, α-linolenic acid and n-3 PUFA in T and TG, docosahexaenoic acid and PUFA in T and arachidonic acid in TG were increased, while saturated fatty acids (SFA) in T were reduced. From 4 days on, PUFA, n-3 PUFA and SFA from T and TG in THY eggs were found to be similar to those of CON eggs. The changes in PL throughout incubation were similar in both dietary treatments. CONCLUSION: Thymol would provide the embryo with PUFA for synthesis/deposition in membranes and/or assign to supply energy. Additionally, thymol supplementation would be advisable for the production of healthier table eggs. © 2017 Society of Chemical Industry.

Assessment of tissue development in fattening quails using the stable isotope technique.

We aimed to study the behaviour of tissue renewal in blood, pectoral muscle and keel of fattening quails during their growth and adulthood through carbon-13 turnover using the stable isotope technique. Three hundred male European quails were randomly subjected to six treatments. Diets fed to animals were initially based on corn and soybean meal and were replaced at 0, 7, 14, 21, 28 and 35 days of age with diets of rice grits and soybean meal. These diets have distinct isotopic signals and over time it changes; the tissue will incorporate dietary signal, and then, we can measure the return time for each tissue in each age. Treatments were T1 (0 to 21 days), T2 (7 to 35 days), T3 (14 to 42 days), T4 (21 to 56 days), T5 (28 to 63 days) and T6 (35 to 70 days). Carbon-13 turnover in tissues of the birds at the above-mentioned time periods was calculated using the first-order exponential function. The values of half-lives were 2.6, 3.6, 5.4, 6.3, 9.0 and 9.8 days for blood and 2.0, 1.7, 4.8, 6.7, 6.9 and 6.6 days for pectoral muscle in treatments T1, T2, T3, T4, T5 and T6 respectively. The half-lives for the keel in treatments T2, T3, T4, T5 and T6 were 2.0, 5.4, 7.7, 8.9 and 15 days respectively. Thus, half-life in sampled tissues generally increased with age of birds. Initially, the tissue half-lives are influenced by growth, and at the end of the growth cycle, the breast muscle tissue was the most metabolically active in either periods, followed by blood and the keel. All tissues showed a sigmoidal growth curve, which can be confirmed by half-life as a function of age.