Structural gonadal lesions observed in Japanese quail (Coturnix coturnix japonica) following exposure during puberty to the neonicotinoid pesticide, imidacloprid.

Exposure to the neonicotinoid insecticide, imidacloprid (IMI), causes reproductive toxicity in mammals and reptiles. However, reports on the effects of IMI on the gonads in birds are grossly lacking. Therefore, this study investigated the effects of pubertal exposure to IMI on the histology, ultrastructure, as well as the cytoskeletal proteins, desmin, smooth muscle actin and vimentin, of the gonads of Japanese quail (Coturnix coturnix japonica). Quails were randomly divided into four groups at 5 weeks of age. The control group was given only distilled water, whereas, the other three experimental groups, IMI was administered by oral gavage at 1.55, 3.1, and 6.2 mg/kg, twice per week for 4 weeks. Exposure to IMI doses of 3.1 and 6.2 mg/kg caused dose-dependent histopathological changes in the ovary and testis. In the ovary, accumulation of lymphocytes, degenerative changes, and necrosis with granulocyte infiltrations were observed, while in the testis, distorted seminiferous tubules, germ cell sloughing, vacuolisations, apoptotic bodies, autophagosomes, and mitochondrial damage were detected. These changes were accompanied by a decreased number of primary follicles (P ≤ 0.05) in the ovary and a decrease (P ≤ 0.05) in the epithelial height, luminal, and tubular diameters of seminiferous tubules at the two higher dosages. In addition, IMI had a negative effect on the immunostaining intensity of desmin, smooth muscle actin, and vimentin in the ovarian and testicular tissue. In conclusion, exposure to IMI during puberty can lead to a range of histopathological alterations in the gonads of Japanese quails, which may ultimately result in infertility.

Effects of Se-enriched yeast on the amelioration of atrazine-induced meat quality degradation.

Atrazine (ATR) is herbicide that causes serious harm to the environment and threatens human food safety. Se-enriched yeast is the best organic selenium source for protecting cells from damage caused by poisonous substances. To explore mechanism of ATR on meat quality degradation and potential protective effects of Se-enriched yeast on ATR-induced muscle injury, quails were treated with ATR and/or Se-enriched yeast for 28 days. The results found ATR disrupted muscle fiber structure and decreased pH, tenderness, water-holding capacity, essential amino acid content and polyunsaturated fatty acid content. ATR aggravated oxidative stress and inflammation by inhibiting Nrf2 pathway and activating NF-κB pathway, ultimately causing apoptosis. However, Se-enriched yeast alleviated ATR-induced alterations in muscle chemical and physical properties by inhibiting oxidative stress and inflammation. Taken together, these results revealed that ATR exposure caused meat quality degradation and Se-enriched yeast had the potential to counteract ATR-induced myotoxicity by inhibiting oxidative stress and inflammation.

Sex-specific effects of dietary restriction on physiological variables in Japanese quails.

Nutritional limitation is a common phenomenon in nature that leads to trade-offs among processes competing for limited resources. These trade-offs are mediated by changes in physiological traits such as growth factors and circulating lipids. However, studies addressing the sex-specific effect of nutritional deficiency on these physiological variables are limited in birds. We used dietary restriction to mimic the depletion of resources to various degrees and investigated sex-specific effects on circulating levels of insulin-like growth factor 1 (IGF-1) and triglycerides in Japanese quails (Coturnix japonica) subjected to ad libitum, 20%, 30% or 40% restriction of their daily requirement, for 2 weeks. We also explored the association of both physiological variables with body mass and egg production. While dietary restriction showed no effects on circulating IGF-1, this hormone exhibited a marked sexual difference, with females having 64.7% higher IGF-1 levels than males. Dietary restriction significantly reduced plasma triglyceride levels in both sexes. Females showed more than six-fold higher triglyceride levels than males. Triglyceride levels were positively associated with body mass in females while showed not association in males. Overall, our findings revealed sex-specific expression of physiological variables under dietary restriction conditions, which coincide with body size.

Dietary restriction and life-history trade-offs: insights into mTOR pathway regulation and reproductive investment in Japanese quail.

Resources are needed for growth, reproduction and survival, and organisms must trade off limited resources among competing processes. Nutritional availability in organisms is sensed and monitored by nutrient-sensing pathways that can trigger physiological changes or alter gene expression. Previous studies have proposed that one such signalling pathway, the mechanistic target of rapamycin (mTOR), underpins a form of adaptive plasticity when individuals encounter constraints in their energy budget. Despite the fundamental importance of this process in evolutionary biology, how nutritional limitation is regulated through the expression of genes governing this pathway and its consequential effects on fitness remain understudied, particularly in birds. We used dietary restriction to simulate resource depletion and examined its effects on body mass, reproduction and gene expression in Japanese quails (Coturnix japonica). Quails were subjected to feeding at 20%, 30% and 40% restriction levels or ad libitum for 2 weeks. All restricted groups exhibited reduced body mass, whereas reductions in the number and mass of eggs were observed only under more severe restrictions. Additionally, dietary restriction led to decreased expression of mTOR and insulin-like growth factor 1 (IGF1), whereas the ribosomal protein S6 kinase 1 (RPS6K1) and autophagy-related genes (ATG9A and ATG5) were upregulated. The pattern in which mTOR responded to restriction was similar to that for body mass. Regardless of the treatment, proportionally higher reproductive investment was associated with individual variation in mTOR expression. These findings reveal the connection between dietary intake and the expression of mTOR and related genes in this pathway.

Dissociating Mechanisms That Underlie Seasonal and Developmental Programs for the Neuroendocrine Control of Physiology in Birds.

Long-term programmed rheostatic changes in physiology are essential for animal fitness. Hypothalamic nuclei and the pituitary gland govern key developmental and seasonal transitions in reproduction. The aim of this study was to identify the molecular substrates that are common and unique to developmental and seasonal timing. Adult and juvenile quail were collected from reproductively mature and immature states, and key molecular targets were examined in the mediobasal hypothalamus (MBH) and pituitary gland. qRT-PCR assays established deiodinase type 2 (DIO2) and type 3 (DIO3) expression in adults changed with photoperiod manipulations. However, DIO2 and DIO3 remain constitutively expressed in juveniles. Pituitary gland transcriptome analyses established that 340 transcripts were differentially expressed across seasonal photoperiod programs and 1,189 transcripts displayed age-dependent variation in expression. Prolactin (PRL) and follicle-stimulating hormone subunit beta (FSHß) are molecular markers of seasonal programs and are significantly upregulated in long photoperiod conditions. Growth hormone expression was significantly upregulated in juvenile quail, regardless of photoperiodic condition. These findings indicate that a level of cell autonomy in the pituitary gland governs seasonal and developmental programs in physiology. Overall, this paper yields novel insights into the molecular mechanisms that govern developmental programs and adult brain plasticity.

The dynamic landscape of chromatin accessibility and active regulatory elements in the mediobasal hypothalamus influences the seasonal activation of the reproductive axis in the male quail under long light exposure.

BACKGROUND: In cold and temperate zones, seasonal reproduction plays a crucial role in the survival and reproductive success of species. The photoperiod influences reproductive processes in seasonal breeders through the hypothalamic-pituitary-gonadal (HPG) axis, in which the mediobasal hypothalamus (MBH) serves as the central region responsible for transmitting light information to the endocrine system. However, the cis-regulatory elements and the transcriptional activation mechanisms related to seasonal activation of the reproductive axis in MBH remain largely unclear. In this study, an artificial photoperiod program was used to induce the HPG axis activation in male quails, and we compared changes in chromatin accessibility changes during the seasonal activation of the HPG axis. RESULTS: Alterations in chromatin accessibility occurred in the mediobasal hypothalamus (MBH) and stabilized at LD7 during the activation of the HPG axis. Most open chromatin regions (OCRs) are enriched mainly in introns and distal intergenic regions. The differentially accessible regions (DARs) showed enrichment of binding motifs of the RFX, NKX, and MEF family of transcription factors that gained-loss accessibility under long-day conditions, while the binding motifs of the nuclear receptor (NR) superfamily and BZIP family gained-open accessibility. Retinoic acid signaling and GTPase-mediated signal transduction are involved in adaptation to long days and maintenance of the HPG axis activation. According to our footprint analysis, three clock-output genes (TEF, DBP, and HLF) and the THRA were the first responders to long days in LD3. THRB, NR3C2, AR, and NR3C1 are the key players associated with the initiation and maintenance of the activation of the HPG axis, which appeared at LD7 and tended to be stable under long-day conditions. By integrating chromatin and the transcriptome, three genes (DIO2, SLC16A2, and PDE6H) involved in thyroid hormone signaling showed differential chromatin accessibility and expression levels during the seasonal activation of the HPG axis. TRPA1, a target of THRB identified by DAP-seq, was sensitive to photoactivation and exhibited differential expression levels between short- and long-day conditions. CONCLUSION: Our data suggest that trans effects were the main factors affecting gene expression during the seasonal activation of the HPG axis. This study could lead to further research on the seasonal reproductive behavior of birds, particularly the role of MBH in controlling seasonal reproductive behavior.

Black soldier fly (Hermetia illucens) larvae meal improves quail growth performance.

The aim of the present study was to determine the nutritional value of black soldier fly (BSF) larvae meal for quail (experiment I) and the dose-response effects of BSF levels on growth performance, relative organ weight, and body composition of growing quails (experiment II). In experiment I, 100 35-day-old quail were distributed in a completely randomized design, with two treatments (reference and test diet) and 10 replicates. The experimental period consisted of 5 days of adaptation, followed by 5 days of total excreta collection. The experimental feed consisted of a reference diet and a test diet formulated with 850 g/kg reference diet and 150 g/kg BSF. In experiment II, 1000 1-day-old quail were distributed in a completely randomized design, with five dietary levels of BSF (0, 25, 50, 75, and 100 g/kg). At 42 days of age, birds were slaughtered, and the relative organ weight and body composition were determined. Apparent metabolizable energy values corrected for nitrogen retention of BSF meal were 13.8 MJ/kg. Across the starter (1-14 days) and overall period (1-42 days), increasing BSF levels had a quadratic effect on body weight and body weight gain. Feed conversion ratio was quadratically affected during the starter phase and linearly reduced over the overall period. Additionally, the BSF levels linearly decreased the small intestine's relative weight at 42 days and had a quadratic effect on the rate of protein deposition. We concluded that the inclusion of 100 g/kg BSF meal improves feed conversion ratio for growing quail.

Differential diagnosis of Eimeria species in farmed Japanese quails (Coturnix japonica).

Similarly to poultry industry, coccidiosis may cause significant economic losses also in the commercial quail industry, an emerging sector undergoing uneven development around the world. Although scant and mostly dated, the available literature reports detailed morphological and morphometric features of both oocysts and sporocysts of the Eimeria species hitherto recognized in Japanese quails, i.e. E. tsunodai, E. uzura, E. bateri, and E. fluminensis. Mixed infections are very common in the field and require an accurate differential diagnosis of diverse species of coccidia, identifying the highly pathogenic ones, in particular E. tsunodai (localized in the caeca), and E. uzura (localized in both caeca and small intestine). This goal is hampered by time-consuming laboratory procedures involving highly qualified staff and facilities, and poorly compatible with routine management practices in farmed quails. A supplemental difficulty is represented by the lack of nucleotide sequences available in GenBank. To overcome these issues, copromicroscopic and molecular analyses (amplifying the 18S rRNA region, and the internal transcribed spacers regions ITS1-5.8rRNA-ITS2) were performed on oocysts populations separately isolated from pools of 12 caecal and 12 cloacal contents collected from 240 naturally infected laying Japanese quails. Data on morphological and morphometric features of 1,000 sporulated oocysts were statistically compared, demonstrating the presence of different Eimeria species colonizing the 2 intestinal tracts. This result was also confirmed by PCR and phylogenetic analysis of the 18S rRNA gene. Overall results allowed to hypothesize the presence of E. uzura in our Japanese quails. Although a certain identification at species level was not obtained, the present study demonstrates that reasonable turnaround times of monitoring procedures performed on Japanese quail farms, shedding light on the in vivo and post-mortem differential diagnosis of coccidiosis can be achieved, and provide obvious benefits in disease understanding and control.

Impact of Dietary Probiotics on the Immune and Reproductive Physiology of Pubertal Male Japanese Quail (Coturnix coturnix japonica) Administered at the Onset of Pre-Puberty.

Fertility in males is dependent on the proper production of sperms involving the synchronization of numerous factors like oxidative stress, inflammatory processes, and hormonal regulation. Inflammation associated with oxidative stress is also known to impair sperm function. Nutritional factors like probiotics and prebiotics have the potential benefits to modulate these factors which may enhance male fertility. In the present study, immature male Japanese quail at the beginning of 3rd week were administered with Lactobacillus rhamnosus (L), Bifidobacterium longum (B), and mannan-oligosaccharides (M) through dietary supplementation in individual groups as well as in combinations like LB and MLB. Markers of oxidative stress including SOD and catalase were examined by native PAGE; inflammatory biomarkers (IL-1ß, IL-10, and NFκB), apoptotic markers (caspase 3 and caspase 7), steroidal hormones, and their receptors estrogen receptor alpha (ERα) and beta (ERß) were assessed in testis. The study reveals that dietary supplementation of 1% L, B, and M in combination significantly and positively increases the overall growth of immature male quail specifically testicular weight and gonadosomatic index (GSI). Furthermore, significant improvement in testicular cell size; increased steroidal hormones like testosterone, FSH, and LH levels; increase in SOD, catalase enzymes; decrease in apoptotic factors Caspase 3, Caspase 7 and immune system strength observed indicated by a decrease in expression of IL-1ß, NFκB; and increase of IL-10 in testis when LBM was used in combination. These variations are attributed to the increase in testicular estrogen receptors alpha and beta, facilitated by the neuroendocrine gonadal axis, ultimately leading to improved male fertility. It can be concluded that the dietary supplementation in combination with L, B, and M enhances male fertility in immature quail by increased expression of estrogen receptors via gut microbiota modulation. It also sheds light on the potential use of these nutritional factors in avian species as therapeutic interventions to overcome low fertility problems in quail thereby benefitting the poultry industry.

Metabolic Rate and Egg Production in Japanese Quails Can Be Predicted by Assessing Growth Parameters of Laying Hens.

The aim of the current study was to assess the female metabolic rate and test the hypothesis that there is a relationship between the egg productivity of Japanese quails from eight breeds and their morphometric, or growth, parameters. Parameters measured were body weight (B), volume (V), and surface area (S), as well as the metabolism level expressed by the ratio S/V. The collected egg performance traits were as follows: the number of eggs produced (N), the average egg weight (W), and the total egg mass (M) (i.e., N multiplied by W). To measure the S and V values, a novel technique was developed that takes into account the similarity of the quail's body to an ellipsoid. An analysis of the relationships between productivity indicators allowed us to introduce a new index called the metabolic index, B·S/V, based on all three main growth parameters in quails. Using the values of this index, we were then able to judge indirectly the level of quails' egg productivity. We went on to assess the N, W, and M values, not only depending on the size of the bird's growth parameters but also according to the degree of their changes during quail growth. These changes were expressed as the slope angles of trend lines describing the growth process data. This approach produced more accurate results for predicting the egg productivity in terms of W and M.

The Histopathological and Oxidative Stress Profiles in Japanese Quails (Coturnix japonica) Induced by Dietary Lead.

In their native habitat, avians are exposed to external toxicity factors, the most prominent of which are chemical lead compounds that threaten human and animal health. The goal of this investigation was to estimate the adverse effects of lead acetate (Pb(CH3COO)2 (H2O)3) on the health status of Japanese quail (Coturnix japonica). 18 adult male Japanese quails (Coturnix coturnix japonica) were employed in this investigation. After two weeks of acclimatization, the birds were randomly divided into three groups: the control group received no Pb+2, the Low Dose Group received 50 mg/kg of Pb+2 as lead acetate Pb(CH3COO)2 (H2O)3 in the diet, and the High Dose Group received 100 mg/kg of Pb+2 as lead acetate Pb(CH3COO)2(H2O)3 in the diet, for 30 days. Results showed that the Pb bioaccumulation was recorded at the highest values in the liver compared with the kidney, and as expected, the ranges of the lead accumulation were significantly higher in the animals who received 100 mg/kg Pb compared with animals who received 50 mg/kg Pb and the control group. In the high dose group, serum content showed significantly increased levels (P≤0.05) of aminotransferase enzymes (ALT and AST ), glucose, creatinine, and uric acid levels compared to other groups, while antioxidant enzymes (CAT, GSH, and GSH-PX) levels in the liver and kidney were significantly reduced (P≤0.05). The results showed that the MDA appeared to be significantly increasing (P≤0.05) in the high dose group compared to the other groups. Compared to the low dose and control groups, the high dosage group produced substantial histological abnormalities in the liver and kidney.

Potential of Full-Fat Silkworm-Based Diets for Laying Quails: Performance and Egg Physical Quality.

The present research was conducted to determine the optimal inclusion level of full-fat silkworm chrysalis meal (SWM) into laying quails' diets, focusing on performance traits and egg physical quality. A total of 240 31-day-old female Japanese quails were randomly assigned to four dietary groups (12 replicates/treatment; 5 quails/replicate); quails were initially fed a standard commercial diet for pullets until 63 days of age. When oviposition started, the experimental groups received the following diets: a conventional corn and soybean-based diet (control diet-C) and three other diets, including 4%, 8%, or 12% of full-fat SWM (SWM4, SWM8, SWM12, respectively). Experimental diets were provided until quails reached 119 days of age. Birds displayed satisfactory productive performance throughout the trial. SWM12 and SWM8 had higher (p < 0.001) egg production but also a higher feed conversion ratio compared to C. At the end of the trial, the eggs edible portion increased, and shell weight decreased with increasing the SWM dietary inclusion level (p < 0.001). At the same time, SWM12 displayed an increase in albumen pH (p < 0.05), even though in the normal range for quail egg. Overall, full-fat SWM (up to 12%) can be considered a promising feed ingredient for laying quails, although higher inclusion levels (>8%) require special attention because SWM also contains anti-nutritional factors.

The Effect of In Ovo Administration of Rosemary Essential Oil on Hatchability, Relative Hatching Weight, and Embryo Mortality Rate in Japanese Quail (Coturnix coturnix japonica).

This study aimed to determine the effects of air sac injection of rosemary essential oil at different concentrations in ovo in quail eggs on hatching rate, relative chick weight at hatching, and embryonic mortality rate. A total of 1060 Japanese quail eggs were divided into four groups: negative control (non-injected), positive control (30 µL sterile distilled water/egg), and two treated groups with 1 and 3 µL oil/egg, respectively. The concentration of 3 µL/egg showed a toxic effect on embryonic development, as revealed by the significantly (p = 0.015) higher post-injection mortality rate (18.21%) compared to 1 µL/egg with 8.3%. Furthermore, hatchability was significantly increased (p = 0.0001) with 1 µL/egg compared to 3 µL/egg with 69.1% and 44.48%, respectively. No significant difference was observed between the concentration of 1 µL/egg and the control groups (p = 0.822). Both l and 3 µL essential oil/egg significantly enhanced (p = 0.0001) relative chick weight at hatching by 67.14% and 70.32%, respectively, compared to the control groups. In conclusion, injecting eggs with 1 µL oil/egg showed positive effects both on hatching and relative chick weight. The concentration of 3 µL/egg was revealed to be toxic, with dramatic effects on embryonic survival.

Effect of the dietary inclusion of Camelina sativa cake into quail diet on live performance, carcass traits, and meat quality.

The present research studied the effect of the dietary inclusion of 3 different camelina (Camelina sativa (L.) Crantz) cakes on the live performance, slaughter traits, and breast meat quality of broiler quails (Coturnix japonica). With this purpose, a total of 480 fifteen-day-old broiler quails of both sexes were allocated to 48 cages (12 replicates/treatment, 10 quails/replicate) and received 4 dietary treatments: a control diet (Control), and 3 diets containing 15% of 1 commercial cultivar (Calena), and 2 improved lines (Pearl: low linoleic acid; Alan: low glucosinolates). During the experiment, individual live weight (LW) and cage feed intake were recorded to calculate body weight gain (BWG) and feed conversion ratio (FCR). At 35 d of age, quails were slaughtered, and carcasses were weighed and dissected to compute yields. On breast (pectoralis major muscle) the measurements and analyses considered ultimate pH, L*a*b* color values, proximate composition, oxidative status, cooking loss and WBSF toughness. Broiler quails receiving diets with camelina cakes exhibited mortality and health status similar to the control. However, growth performance was impaired in camelina-fed groups, especially for Calena (P < 0.05). Overall LW and BWG were slightly lower, whereas feed intake was slightly higher in camelina-fed groups compared to Control (P > 0.05). Therefore, higher FCR was recorded for camelina-fed groups compared to Control (P = 0.0004). Moreover, breast meat from Calena treatment displayed higher water (P = 0.0170), and lower lipid (P = 0.0051) contents compared to those of the Control group, while protein and ash content remained unaffected. Heme-iron content and oxidative status of breast meat were not influenced by the dietary incorporation of camelina (P > 0.05). The research outcomes indicated that camelina cakes can be used as an alternative feed ingredient for broiler quails' diets, without compromising carcass yields and meat quality. However, as 15% dietary incorporation worsened live performance, the ideal camelina cake inclusion level should be thoroughly investigated as well as a parallel research effort into further reducing glucosinolates content of camelina.

Genome organization of major tandem repeats and their specificity for heterochromatin of macro- and microchromosomes in Japanese quail.

Tandemly repeated DNAs form heterochromatic regions of chromosomes, including the vital centromeric chromatin. Despite the progress in new genomic technologies, tandem repeats remain poorly deciphered and need targeted analysis in the species of interest. The Japanese quail is one of the highest-producing poultry species as well as a model organism. Its genome differs by a noticeable accumulation of heterochromatin, which led to an increase by 1/7 compared to the chicken genome size. Prominent heterochromatin blocks occupy the short arms of acrocentric macrochromosomes and of microchromosomes. We have applied de novo repeat finder approach to unassembled raw reads of the Japanese quail genome. We identified the 20 most common tandem repeats with the abundance >1 Mb, which represent about 4.8% of the genome. We found that tandem repeat CjapSAT primarily contributes to the centromeric regions of the macrochromosomes CJA1-8. Cjap31B together with previously characterized BglII makes up centromere regions of microchromosomes and W chromosome. Other repeats populate heterochromatin of microchromosomal short arms in unequal proportions, as revealed by fluorescence in situ hybridization. The Cjap84A, Cjap408A, and CjapSAT repeat sequences show similarities to retrotransposon motifs. This suggests that retroelements may have played a crucial role in the distribution of repeats throughout the Japanese quail genome.

Novel Single Nucleotide Polymorphisms (SNPs) and Genetic Features of the Prion Protein Gene (PRNP) in Quail (Coturnix japonica).

Prion diseases are fatal infectious diseases caused by conformational changes of a prion protein (PrPSc) derived from a normal prion protein (PrPC). Prion diseases have been reported in several mammalian hosts but not in any birds, including the most popular poultry species, of which chickens showed some resistance to experimental prion infection. To identify the genetic polymorphisms in the quail prion protein gene (PRNP), polymerase chain reaction and DNA sequencing were performed with gene-specific primers in 164 quails. Four in silico programs, including PROVEAN, PANTHER, SIFT, and AMYCO, were used to investigate the effect of non-synonymous single nucleotide polymorphisms (SNPs) on quail PrP. Furthermore, to investigate the genetic relationship of avian PrPs, phylogenetic analysis and multiple sequence alignments were performed using MEGA X program. Finally, the secondary and tertiary structures of avian PrPs were analyzed by SWISS-MODEL. We identified 33 novel SNPs in the quail PRNP gene, including three non-synonymous SNPs, c.56C>T (T19I), c.60C>T (V21I), and c.61G>A (A22S). Although V21I was predicted to have deleterious effects by SIFT, the substitutions of all three amino acids did not affect the amyloid propensity, 3D structure, or hydrogen bonds of quail PrP. Quail PrP showed a close evolutionary relationship and similar secondary and tertiary structures to chicken PrP compared to duck PrP. To our knowledge, this is the first report on the genetic and structural properties of the quail PRNP gene.

A Novel Egg-In-Cube System Enables Long-Term Culture and Dynamic Imaging of Early Embryonic Development.

The avian egg is a closed system that protects the growing embryo from external factors but prevents direct observation of embryo development. Various culture systems exist in the literature to study the development of the embryo for short periods of incubation (from 12 h up to a maximum of 60 h of egg incubation). A common flaw to these culture techniques is the inability to culture the unincubated avian blastoderm with intact tissue tensions on its native yolk. The goal of this work is to create a unique novel egg-in-cube system that can be used for long-term quail embryo culture initiated from its unincubated blastoderm stage. The egg-in-cube acts as an artificial transparent eggshell system that holds the growing embryo, making it amenable to microscopy. With the egg-in-cube system, quail embryos can be grown up to 9 days from the unincubated blastoderm (incubated in air, 20.9% O2), which improves to 15 days on switching to a hyperoxic environment of 60% O2. Using transgenic fluorescent quail embryos in the egg-in-cube system, cell movements in the unincubated blastoderm are imaged dynamically using inverted confocal microscopy, which has been challenging to achieve with other culture systems. Apart from these observations, several other imaging applications of the system are described in this work using transgenic fluorescent quail embryos with upright confocal or epifluorescence microscopy. To demonstrate the usefulness of the egg-in-cube system in perturbation experiments, the quail neural tube is electroporated with fluorescent mRNA "in cubo", followed by the incubation of the electroporated embryo and microscopy of the electroporated region with the embryo in the cube. The egg-in-cube culture system in combination with the "in cubo" electroporation and dynamic imaging capabilities described here will enable researchers to investigate several fundamental questions in early embryogenesis with the avian (quail) embryo on its native yolk.

Effect of the Prolactin Inhibitor Cabergoline and the Gonadotropin Releasing-Hormone Agonist Deslorelin in the Suppression of Plasma Prolactin Concentrations and Egg Laying in Quail (Coturnix japonica).

Egg binding and excessive laying frequently affect avian patients, and in many cases the treatment includes suppression of egg production. Currently, for the suppression of egg production in avian patients, a gonadotropin-releasing hormone agonist, in the form of a deslorelin implant, is often used. However, the commercially available deslorelin implants have an undesired delayed onset, as well as a potential brief increase in gonadotropin secretion after administration ("flare-up" effect) that can lead to oviposition before the actual suppression of gonadotropins. The objective of this study was to investigate whether the prolactin inhibitor cabergoline suppresses ovulation and whether it could be used to bridge the time until the onset of effect by the deslorelin implant. We measured the effect of cabergoline (30 µg/kg PO q24h × 14 days), deslorelin implants (4.7 mg SC), and a combination of both on egg laying and plasma prolactin concentrations in 37 quail (Coturnix japonica) over 6 weeks. Quail were divided into 4 groups: group DesCab (deslorelin implant and cabergoline oral; n = 9); group DesPlac (deslorelin implant and placebo oral; n = 9); group PlacCab (placebo implant and cabergoline oral; n = 9); and group PlacPlac (placebo implant and placebo oral; n = 10). Regular egg laying stopped in 100% (9/9) of birds in group DesCab and 78% (7/ 9) of birds in group DesPlac within 5 days of placing the deslorelin implant. No bird ceased egg production in group PlacCab (0/9), and 10% of birds ceased egg production intermittently in group PlacPlac (1/10). Treatment with the deslorelin implant (P < .001) and with cabergoline (P = .04) had a significant (negative) influence on plasma prolactin concentrations compared with the baseline. The interaction of deslorelin and cabergoline treatment, as well as time after initiation of treatment, did not have a significant effect on plasma prolactin concentrations. These results show that daily oral cabergoline has no significant influence on egg laying and only a minor biologically nonsignificant effect on lowering the relative plasma prolactin concentrations in quail.

Arginine requirement for egg production in Japanese quail.

The objective of this study was to determine the ideal arginine intake for egg production in Japanese quail using the dilution technique. A completely random design was used, with 8 treatments (seven concentrations of arginine plus a control diet) and ten replicates, totaling 80 Japanese quails. The digestible arginine levels included in the study were 0.361%, 0.603%, 0.843%, 1.084%, 1.204%, 1.311%, and 1.460%. The variables analyzed were feed intake, egg production (EP), egg weight, egg output (EO), feed conversion ratio, and body weight were performed using a mixed model. When the effect of arginine levels (P ≤ 0.05) was detected, the model's broken line linear-plateau (BL), quadratic-plateau (BLq), and the first intercept of the BLq in the plateau of BL were adjusted to determine the ideal arginine intake. It observed that the arginine levels modified the quail responses (P < 0.001). Egg production was 10% with the 0.361% arginine in diet and recovered (97%) with the 1.311% arginine diet. The BL and BLq models estimated 232 mg/quail per day and 351 mg/quail per day for EO, respectively. The first intercept obtained was 290 mg/quail per day or 1,411%, which was considered the optimum level arginine intake for EO in Japanese quail.

Short communication: Effects of in-ovo injection of endocrine disruptors and methyltransferase inhibitor on quail growth and egg-laying performances.

Early experiences, including prenatal environment, are known to influence a wide variety of mechanisms involved in the phenotype elaboration. We investigated the effect of the addition of endocrine disruptors or of a methyltransferase inhibitor during the embryonic development of quails from different genetic backgrounds (four different quail lines) on their growth and egg-laying performances. Fifty-four pairs of parents per line were used and fertilised eggs from each pair were randomly divided into five groups: a control group without any injection, an injected control group treated by injection into the egg of sesame oil, and three groups treated by injection of Genistein, Bisphenol A or 5-Aza-2'-deoxycytidine. All quails were individually weighed at 8, 21, 36 and 78 days. The age at first egg laid and the number of eggs laid were recorded. These analyses revealed a significant impact of the treatment on growth but no influence on the egg-laying traits. All three molecules significantly affected at least one of the analysed growth traits. In conclusion, we showed that the injection of endocrine disruptors or DNA methyltransferase inhibitor into the egg had significant effects on quail development; these effects were specific to each treatment, but no interaction between line and treatment was observed.