Aquaporin regulates cell rounding through vacuole formation during endothelial-to-hematopoietic transition.

Endothelial-to-hematopoietic transition (EHT) is crucial for hematopoietic stem cell (HSC) generation. During EHT, the morphology of hemogenic endothelial cells (HECs) changes from flat and adherent to spherical hematopoietic cells, which detach from the dorsal aorta. HECs attain a rounded shape in a mitosis-independent manner before cell adhesion termination, suggesting an atypical cell-rounding mechanism. However, the direct mechanisms underlying this change in cell morphology during EHT remain unclear. Here, we show that large vacuoles were transiently formed in avian HECs, and that aquaporin 1 (AQP1) was localized in the vacuole and plasma membranes. Overexpression of AQP1 in non-HECs induced ectopic vacuole expansion, cell rounding and subsequent cell detachment from the endothelium into the bloodstream, mimicking EHT. Loss of redundant AQP functions by CRISPR/Cas9 gene editing in HECs impeded the morphological EHT. Our findings provide the first evidence to indicate that morphological segregation of hematopoietic cells from endothelial cells is regulated by water influx into vacuoles. These findings provide important insights for further exploration of the mechanisms underlying cell/tissue morphogenesis through water-adoptive cellular responses.

A novel qPCR-based technique for identifying avian sex: An illustration within embryonic craniofacial bone.

Sex is a biological variable important to consider in all biomedical experiments. However, doing so in avian embryos can be challenging as sex can be morphologically indistinguishable. Unlike humans, female birds are the heterogametic sex with Z and W sex chromosomes. The female-specific W chromosome has previously been identified in chick using a species-specific polymerase chain reaction (PCR) technique. We developed a novel reverse transcription quantitative PCR (RT-qPCR) technique that amplifies the W chromosome gene histidine triad nucleotide-binding protein W (HINTW) in chick, quail, and duck. Accuracy of the HINTW RT-qPCR primer set was confirmed in all three species using species-specific PCR, including a novel quail-specific HINTW PCR primer set. Bone development-related gene expression was then analyzed by sex in embryonic lower jaws of duck and quail, as adult duck beak size is known to be sexually dimorphic while quail beak size is not. Trends toward sex differences were found in duck gene expression but not in quail, as expected. With these novel RT-qPCR and PCR embryo sexing methods, sex of chick, quail, and duck embryos can now be assessed by either/both RNA and DNA, which facilitates analysis of sex as a biological variable in studies using these model organisms.

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.

Clinical Features of Quail Egg Ingestion in Patients with Acquired Tolerance to Hen Eggs: A Case Series Study.

INTRODUCTION: Patients with hen's egg allergy are often instructed to avoid consuming other avian eggs, such as quail eggs. However, it is unclear whether patients with an acquired tolerance to hen eggs continue to avoid consuming quail eggs. This study aimed to evaluate the clinical features of quail egg ingestion. METHODS: This prospective case series included children aged ≥1 year with hen's egg allergy who were recruited between October 2019 and February 2021 in our hospital. We conducted an oral food challenge (OFC) with three boiled quail eggs to evaluate the clinical features of quail egg ingestion in patients with acquired tolerance to hen eggs. The primary outcome was a positive OFC after ingesting three quail eggs. Secondary outcomes were cross-antigenicity between hen and quail eggs observed through the skin prick test (SPT) and pattern of quail egg allergy, comprising the onset of reaction, and severity. The correlation between the diameters of the wheals with SPT in hen and quail eggs was evaluated using the Pearson product-moment correlation coefficient. RESULTS: A total of 62 patients underwent the quail egg OFC. The median (interquartile range) age of the participants was 3 (2-5) years. Thirty-three (53%) patients had a history of anaphylaxis due to hen eggs. The median total immunoglobulin E (IgE) level in patients who underwent the OFC with half a heated whole hen's egg was 271 (98-593) IU/mL. The median specific IgE level in egg white and ovomucoid was 9.7 (3.2-21.5) and 4.4 (1.3-6.9) UA/mL, respectively. The quail egg OFC results revealed that none of the 59 patients who ate the three quail eggs completely had an allergic reaction. The SPT-positive and SPT-negative rates in raw and boiled hen and quail egg whites were both correlated. The diameters of wheals with SPT in raw hen and quail egg whites and yolks were positively correlated. CONCLUSION: Patients with an acquired tolerance to hen eggs may not be required to avoid consuming quail eggs.

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.

Plasma activated water effects on behavior, performance, carcass quality, biochemical changes, and histopathological alterations in quail.

BACKGROUND: Plasma-activated water (PAW) is an innovative promising technology which could be applied to improve poultry health. The current study investigated the effects of drinking water supply with PAW on quail behaviour, performance, biochemical parameters, carcass quality, intestinal microbial populations, and internal organs histopathology. A total of 54 twenty-one-day-old Japanese quail chicks were randomly allotted to three treatments provided with PAW at doses 0, 1 ml (PAW-1), and 2 ml (PAW-2) per one litter drinking water. Each treatment contained 6 replicates (3 birds/ cage; one male and two females). RESULTS: The results clarified that there were no significant (P > 0.05) changes in behaviour, and performance. For the biochemical indicators, the PAW-1 group showed significantly higher serum H2O2, total protein and globulin levels compared with the other groups (P = 0.015, < 0.001, and 0.019; respectively). PAW groups had significantly lower serum creatinine and urea levels than the control (P = 0.003). For the carcass quality, the internal organs relative weight between different treatments was not changed. In contrast, there was a significant increase in the meat colour, taste, and overall acceptance scores in PAW groups compared with the control one (P = 0.013, 0.001, and < 0.001; respectively). For the intestinal microbial population, lactobacilli count was significantly higher in PAW-2 compared with the control group (P = 0.014), while there were no changes in the total bacterial count between different treatment groups. Moreover, mild histological changes were recorded in the intestine, liver, and spleen of PAW groups especially PAW-2 compared with the control one. CONCLUSIONS: PAW offered benefits, such as reducing creatine and urea levels, improving meat characteristics, and increasing lactobacilli count, all of which are crucial for sustainable quail farming. Therefore, further research is needed.

Characteristics of intestinal bacteriophages and their relationship with Bacteria and serum metabolites during quail sexual maturity transition.

BACKGROUND: Bacteriophages are prokaryotic viruses that rank among the most abundant microbes in the gut but remain among the least understood, especially in quails. In this study, we surveyed the gut bacteriophage communities in 22 quails at different ages (days 20 and 70) using shotgun metagenomic sequencing. We then systematically evaluated the relationships with gut bacteria and host serum metabolites. RESULTS: We discovered that Myoviridae and Siphoviridae were the dominant bacteriophage families in quails. Through a random forest and LEfSe analysis, we identified 23 differential bacteriophages with overlapping presence. Of these, 21 bacteriophages (e.g., Enterococcus phage IME-EFm5 and Enterococcus phage IME-EFm1) showed higher abundances in the day 20 group, while two bacteriophages (Bacillus phage Silence and Bacillus virus WPh) were enriched in the day 70 group. These key bacteriophages can serve as biomarkers for quail sexual maturity. Additionally, the differential bacteriophages significantly correlated with specific bacterial species and shifts in the functional capacities of the gut microbiome. For example, Enterococcus phages (e.g., Enterococcus phage EFP01, Enterococcus phage IME-EFm5, and Enterococcus phage IME-EFm1) were significantly (P < 0.001, FDR) and positively correlated with Enterococcus faecalis. However, the relationships between the host serum metabolites and either bacteriophages or bacterial species varied. None of the bacteriophages significantly (P > 0.05, FDR) correlated with nicotinamide riboside and triacetate lactone. In contrast, some differential bacterial species (e.g., Christensenella massiliensis and Bacteroides neonati) significantly (P < 0.05, FDR) correlated with nicotinamide riboside and triacetate lactone. Furthermore, characteristic successional alterations in gut bacteriophages, bacteria, and host serum metabolites across different ages highlighted a sexual maturity transition coexpression network. CONCLUSION: This study improves our understanding of the gut bacteriophage characteristics in quails and offers profound insights into the interactions among gut bacteriophages, bacteria, and host serum metabolites during the quail's sexual maturity transition.

Early-life group size influences response inhibition, but not the learning of it, in Japanese quails.

In complex social environments, animals benefit from suppressing inappropriate responses (known as Response Inhibition) to avoid conflicts and maintain group cohesion. Recent research suggests that an individual's early-life social environment can shape their response inhibition. However, these findings have mostly been correlational, and results vary across species. Furthermore, the role of learning is often overlooked when measuring response inhibition, despite its potential importance to understanding group differences. We investigated the effect of early-life group size, a key determinant of social complexity, on response inhibition in Japanese quails (Coturnix japonica), whilst taking the role of learning into account. Quails (n = 120) were raised in either small groups of five or large groups of 15 individuals. Response inhibition was assessed using the cylinder task. Up to ten trials were administered to assess whether the birds' responses changed with increasing experience of the task. Among the quails that completed ten trials, we found that those raised in large groups consistently spent less time pecking the cylinder than those raised in small groups. The quails' responses were also influenced by their body condition, food motivation and sex. Importantly, the quails learned to inhibit their responses - successful trials increased, and time spent pecking the cylinder decreased, across ten trials. However, learning rates did not differ between the treatment groups. These findings suggest that early-life social group size promotes the development of response inhibition in quails, but not their learning of it, during the cylinder task.

RNA sequencing analysis reveals that missense mutation in SOX10 is associated with iris color phenotype in quail.

To investigate the molecular mechanisms underlying the differences in iris color in quail, the transcriptome of iris tissue from black quail and Korean quail at day 10 of hatching was RNA sequenced in this study. The differentially expressed genes (DEGs) were screened, functionally annotated and enriched after the quality control and mapping of the raw data. RT-qPCR validation was performed using EIF2S3 as an internal reference gene. The screened SNPs were studied by bioinformatics analysis and iris color correlation analysis. The results showed that there were 425 upregulated genes and 364 downregulated genes in 789 DEGs. Gene Ontology (GO) enrichment analysis revealed that 139 DEGs were significantly enriched in 154 GO terms. The Kyoto Encyclopedia of Genes and Genomes enrichment results showed that the Notch signaling pathway, melanogenesis and tyrosine metabolism were associated with pigment synthesis (p < 0.05). The expression levels of the ASIP, MLPH, PMEL, TYR and SOX10 genes were significantly different in black quail iris and Korean quail iris, as verified by RT-qPCR. The SOX10 gene c.324G>C mutation, which caused the replacement of p.Glu108Asp, had a highly significant correlation with iris color in black quail and Korean quail, which may be one of the reasons for different in iris color between these two quail species.

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.

Potential effect dietary supplementation of calcium tetraborate in quails exposed to cadmium: Its impact on productive performance, oxidative stress, cecal microflora, and histopathological changes.

Cadmium (Cd) is a ubiquitous environmental pollutant, and Cd exposure harms human health, agriculture, and animal husbandry. The present study aimed to investigate the potential protective effect of dietary supplementation of calcium tetraborate (CTB) on productive performance, oxidative stress, cecal microflora, and histopathological changes in quail exposed to Cd. A total of one hundred twenty, 6-week-old Japanese quail (four females and two males/replicate) were divided into four groups (30 quails/group): the control group (feeding basic diet), CTB group (basic diet containing 300 mg/kg CaB4O7, 22.14% elemental B/kg diet), the Cd group (basic diet containing 100 mg/kg cadmium chloride (CdCl2) (total Cd content of 92.1 mg/kg)) and the CTB + Cd group (basic diet containing 300 mg/kg CTB and 100 mg/kg CdCl2). The results showed that Cd exposure caused decreased performance, increased the proportion of broken and soft-shelled eggs, induced oxidative stress, affected cecal microflora, epicardial hemorrhages in the heart, focal necrosis in the liver, degeneration in the kidneys, and degenerated and necrotic seminiferous tubules in the testicles. CTB prevented Cd-induced oxidative stress in liver tissue by increasing total antioxidant status and reducing total oxidant status. In addition, CTB improved egg production and feed conversion ratio (FCR). CTB protected the cecal microflora by inhibiting Enterobacteriaceae and promoting Lactobacillus. CTB also reduced Cd-induced histopathological damage in the heart, liver, kidneys, and testicles. In conclusion, these findings suggest that CTB could be used in Cd-challenged quail, and this compound provides new insights into the toxicity of environmental Cd.

Age-related changes in testicular histological structure, testosterone level, libido and semen traits in male Coturnix quail.

To investigate the effect of age in male quail on testicular weight and histology, sexual libido and semen characteristics, a study was performed on 100 quails at 10, 16, 22, 28 and 34 weeks of age. The body and testicular weights were significantly (p < .05) higher at 16 and 22 than at 28 weeks of age. The circumference and diameter of the seminiferous tubules were significantly (p < .05) higher at 28 and 34 than at 10 and 16 weeks of age. Histological evaluation of testicular slices revealed advanced and effective seminiferous tubes as early as 10 weeks, while spermatogenic activity peaked at 16 weeks of age. The highest semen volume, sperm motility and sperm concentration were observed at weeks of age and then decreased gradually with age. At the same time, the testosterone level and libido were significantly (p < .05) higher at 22 than at 10 weeks of age. Furthermore, the age was positive (p < .05), correlated to seminiferous tubule circumference and diameter, and negative (p < .05), correlated to sperm concentration. In conclusion, the age of quail markedly affected the testicular histological structure, libido, testosterone level and semen characteristics.

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.

Glycerol is not suitable for the cryopreservation of quail semen.

1. This study investigated the potential use of glycerol as a cryoprotectant for quail sperm cells. Its role in maintaining sperm fertilising ability in vivo and in vitro quality parameters, such as motility, was assessed.2. The data showed that the presence of glycerol in semen samples was associated with infertility, which suggested that removal prior to insemination is mandatory. Removal through serial dilution centrifugation was associated with fewer than 5% of motile sperm cells and resulted in no fertility.3. In conclusion, glycerol alone is not suitable for quail semen cryopreservation, and other approaches need to be investigated to develop cryobanking programmes for this species.

Dietary ratio of linoleic to alpha-linolenic acid affects the reproductive performance of Japanese quail.

1. This study was conducted to assess the effects of different dietary omega 6:3 ratios fed to male and female Japanese quail breeders on incubation performance, chick quality and progeny performance.2. A completely randomised design was used, with five diets containing different ratios of vegetable oils rich in linoleic acid (LA from soybean oil) or α-linolenic acid (ALA from linseed oil) with LA/ALA ratios of 13.75:1, 10.69:1, 7.63:1, 4.57:1 and 1.48:1 with 12 cage replicates containing six birds each.3. There was a quadratic effect of the LA/ALA ratio on total hatchability (p < 0.011), fertile hatchability (p = 0.046) and total mortality (p = 0.046). There was no effect on fertility (p > 0.05). The LA/ALA ratios of 1.48 and 13.75 fed to both hens and cockerels or hens resulted in greater fertility, as measured by the number of days after copulation during which fertile eggs were laid and the number of points of hydrolysis on the perivitelline membrane. A decreasing linear effect (p < 0.0001) was observed on chick length and an increasing linear effect on body weight at 1 day of age. There were no effects on progeny performance.4. The LA/ALA ratio affected yolk mineral matter (p = 0.009), crude protein (p = 0.091), chick mineral matter (p < 0.038) and ether extract (p < 0.0001) contents. Maternal diet affected the fatty acid profile of egg yolk and chick liver, indicating that dietary contents were transferred to eggs and chicks.5. Fertile egg production increased with lower LA/ALA ratios. Therefore, linseed oil can be used together with soybean oil to formulate diets for female Japanese quail obtain LA/ALA ratios between 4:1 and 10:1.

The relationship of MITF gene expression and promoter methylation with plumage colour in quail.

1. This study focused on the relationship between MITF mRNA expression and plumage colour in quail and the effect of promoter methylation on the expression of MITF mRNA.2. The CDS region of MITF mRNA was cloned by RT-PCR, followed by DNA sequencing. The RT-qPCR method was used to analyse the expression levels of MITF mRNA in dorsal skin tissue in Korean quail and Beijing white quail. The promoter region of the MITF gene was cloned, and the CpG island was predicted by the CpGplot program. The methylation levels of the CpG island were analysed using BS-PCR technology.3. Quail MITF mRNA contains a 1,476 bp complete ORF, which encodes a 492 amino acid residue protein. The MITF protein has no signal peptide or transmembrane region. The expression of MITF mRNA in dorsal tissue of Korean quail was significantly higher than that in Beijing white quail (p < 0.01). Abundant cis-elements and a 346 bp CpG island were found in the promoter region of the MITF gene. The average methylation level of the CpG island was 22 (22%) in Korean quail, and 46 (30%) in Beijing white quail (p < 0.05).4. The hypermethylation of the MITF gene promoter region in Beijing white quail resulted in a decrease in expression level, which was related to white feather colour.

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.

Effectiveness of typodont, quail egg and virtual simulation for ultrasonic periodontal scaling teaching among pre-clinical students: a randomized trial.

BACKGROUND: This study aimed to compare the efficacy of three different techniques, namely virtual simulation technology (VS), traditional pathological typodont (TT), and quail egg (QE), in pre-clinical training of periodontal ultrasonic scaling. It also aimed to propose an integrated teaching approach for ultrasonic scaling teaching. METHODS: This single-blind randomized multi-arm trial enrolled 108 fourth-year students from Guanghua School of Stomatology at Sun Yat-sen University. The participants were randomly, evenly assigned to VS, TT, or QE group. First, the participants received theoretical review on ultrasonic scaling and demonstrative teaching. Then in the 90-minute operation training by group, students used traditional typodont equipped in head-simulators, raw quail eggs, or scaling module of the UniDental VS system respectively. Then all participants practiced on pathological models for 30 min. In the final operation examination, participants were instructed to remove the supra- and sub-gingival calculi pre-set on designated teeth by ultrasonic scalers within 30 min. Their performances were evaluated by residual calculus rate and a multi-perspective scoring scale. After the examination, questionnaires were provided to assess the teaching effects of each method and the fidelity of VS. Statistical analysis was carried out using one-way, two-way ANOVA, and multiple t-test. RESULTS: Students in VS group had significant higher total test scores than QE group (87.89 ± 6.81, 83.53 ± 8.14) and TT group (85.03 ± 6.81). VS group scored higher in several dimensional comparisons with the other two groups, especially in difficult situations. QE group had higher scores particularly in force application and supra-gingival scaling. TT group scored the highest in pivot stability practice and body position training. Students gave higher scores when assessing the fidelity of VS than experienced teachers. CONCLUSION: The study highlights the importance of specialized pre-clinical training on ultrasonic scaling for dental students. The methods adopted in current study (VS, TT and QE) each offered unique advantages in education, which can be combined to create an integrative teaching procedure. This procedure aims to provide an effective, advisable and normative pre-clinical training procedure for ultrasonic scaling. By utilizing the strengths of each method, dental educators can deliver high-quality training and ensure that students are well-prepared for clinical practice.

Dietary supplementation with mushroom powder (Agaricus bisporus) on performance, carcass traits, meat quality, and bone biomechanical properties of quail (Coturnix coturnix japonica).

This study was performed to determine the effect of mushroom powder (MP) (Agaricus bisporus) supplementation on growing Japanese quail (Coturnix coturnix japonica). A total of 300 unsexed 1-day-old Japanese quails with similar body weights (8.38 ± 0.2 g) were randomly assigned to five treatment groups with six replications. Additions of 0, 0.25, 0.50, 0.75, or 1.00% of MP to the basal diet were used to develop the treatment groups. Quails were fed ad libitum for 42 days. At the end of the experiment, 12 quails from each experimental unit were euthanised to determine performance, carcass traits, meat quality, and bone biochemical properties. Results showed that all dietary MP did not negatively affect any performance parameters (P > 0.05), while by the third week of life, there was an increase (P < 0.05) in body weight and body weight gain in the quails of the 0.75% MP group compared to the control group. Nevertheless, these differences disappeared at the end of the trial (P > 0.05). No differences were observed (P > 0.05) for any of the studied carcass traits, except for the pancreas weight which decreased (P < 0.05) with the addition of high MP (1.00%). Regarding meat quality, all color parameters were affected on the fifth day of sampling (P < 0.05) but not on the first day (P > 0.05). It was detected that the breast of the quails in group 0.75% MP had the highest L* value and the lowest a* value. While the breast of the 1.00% MP group had the highest b value and the lowest pH value. Dietary MP enhanced oxidative stability, reducing malondialdehyde (MDA) value in the breast compared to the control at both sampling points (P < 0.01), being more noted on the fifth day of sampling. Bone biomechanical properties (in terms of shear force or shear stress) were improved (P < 0.01) with the dietary addition of MP at 0.75% compared to the control. It can be suggested that MP is a secure ingredient in animal feed without negatively affecting performance parameters, carcass traits, or meat quality. Therefore, including an interval of 0.50-0.75% of MP in the diet of growing quails could be a suitable strategy to improve certain parameters such as the meat's oxidative stability and the bone's biomechanical parameters. Moreover, the efficacy of MP on performance development would be greater during the first weeks of the quails' life due to their intestinal conditions at this stage.

Expression atlas of avian neural crest proteins: Neurulation to migration.

Neural crest (NC) cells are a dynamic population of embryonic stem cells that create various adult tissues in vertebrate species including craniofacial bone and cartilage and the peripheral and enteric nervous systems. NC development is thought to be a conserved and complex process that is controlled by a tightly-regulated gene regulatory network (GRN) of morphogens, transcription factors, and cell adhesion proteins. While multiple studies have characterized the expression of several GRN factors in single species, a comprehensive protein analysis that directly compares expression across development is lacking. To address this lack in information, we used three closely related avian models, Gallus gallus (chicken), Coturnix japonica (Japanese quail), and Pavo cristatus (Indian peafowl), to compare the localization and timing of four GRN transcription factors, PAX7, SNAI2, SOX9, and SOX10, from the onset of neurulation to migration. While the spatial expression of these factors is largely conserved, we find that quail NC cells express SNAI2, SOX9, and SOX10 proteins at the equivalent of earlier developmental stages than chick and peafowl. In addition, quail NC cells migrate farther and more rapidly than the larger organisms. These data suggest that despite a conservation of NC GRN players, differences in the timing of NC development between species remain a significant frontier to be explored with functional studies.