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25 pages, 2866 KiB  
Review
Heat Stress in Japanese Quails (Coturnix japonica): Benefits of Phytase Supplementation
by Apolônio Gomes Ribeiro, Raiane dos Santos Silva, Dayane Albuquerque da Silva, Júlio Cézar dos Santos Nascimento, Lilian Francisco Arantes de Souza, Edijanio Galdino da Silva, José Evangelista Santos Ribeiro, Danila Barreiro Campos, Clara Virgínia Batista de Vasconcelos Alves, Edilson Paes Saraiva, Fernando Guilherme Perazzo Costa and Ricardo Romão Guerra
Animals 2024, 14(24), 3599; https://doi.org/10.3390/ani14243599 - 13 Dec 2024
Viewed by 767
Abstract
In tropical and subtropical climate regions, heat stress is one of the main causes of production losses in laying quails, aggravated by the antinutritional effects of the phytate in diet ingredients, which negatively affect the bioavailability of minerals, especially calcium and phosphorus. This [...] Read more.
In tropical and subtropical climate regions, heat stress is one of the main causes of production losses in laying quails, aggravated by the antinutritional effects of the phytate in diet ingredients, which negatively affect the bioavailability of minerals, especially calcium and phosphorus. This situation results in a reduction in production and the quality of eggs from commercial laying quails. Several nutritional strategies are utilized to reduce the adverse effects of high temperatures and antinutritional factors such as phytate. Among these strategies, the use of exogenous enzymes, such as phytase, stands out as a viable alternative. Phytase breaks down phytate molecules, optimizing the absorption of essential minerals and improving productive performance and egg quality under unfavorable conditions. Specifically, it is recommended to use 1500 FTU of phytase, as it not only reduces the effects of thermal stress but also enhances eggshell thickness and calcium absorption. In this context, this bibliographic review sought to produce a document demonstrating the beneficial effects of the phytase enzyme on the hydrolysis of the phytate molecule, the availability of calcium for Japanese quails (Coturnix japonica), and its implications in thermal stress due to heat. Full article
(This article belongs to the Section Animal Nutrition)
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<p><span class="html-italic">Myo-inositol</span> (<b>a</b>) and myo-inositol 1,2,3,4,5,6-hexakis dihydrogen phospodium (InsP6) (<b>b</b>).</p>
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<p>Hydrolysis of the phytate molecule through the action of the phytase enzyme: I (inositol); P (phosphate); IP6 (myo-inositol hexakisphosphate); IP5 (Myo-inositol pentachysphosphate); IP4 (Myo-inositol tetrakisphosphate); IP3 (Myo-inositol triphosphate); IP2 (Myo-inositol bisphosphate); and IP1 (Myo-inositol monophosphate).</p>
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<p>Classification of phytases.</p>
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<p>Schematic drawing of transepithelial calcium transport. Integrated model of active epithelial Ca<sup>2+</sup> transport. Ca<sup>2+</sup> enters the cell from the luminal side via TRPV6, subsequently binds to calbindin-D28K, and is extruded at the basolateral membrane by a Na+/Ca<sup>2+</sup> exchanger (NCX1) and/or a plasma membrane Ca<sup>2+</sup>-ATPase (PMCA1b). The active form of vitamin D [1,25(OH)2D3] stimulates the individual steps of transcellular Ca<sup>2+</sup> transport by increasing the expression level of TRPV6, calbindin-D28K and the extrusion systems. TRPV6 (transient receptor potential vanilloid channel type 6); VDR (vitamin D receptor).</p>
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<p>Calcium selective channel TRPV6 and Calbindin-D28K. TRPV6 (transient receptor potential vanilloid channel type 6); VDR (vitamin D receptor).</p>
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18 pages, 4682 KiB  
Article
Screening Algal and Cyanobacterial Extracts to Identify Potential Substitutes for Fetal Bovine Serum in Cellular Meat Cultivation
by Nikolina Sibinčić, Maja Krstić Ristivojević, Nikola Gligorijević, Luka Veličković, Katarina Ćulafić, Zorana Jovanović, Aleksandar Ivanov, Lora Tubić, Carole Vialleix, Thibaut Michel, Tatjana Srdić Rajić, Milan Nikolić, Marija Stojadinović and Simeon Minić
Foods 2024, 13(23), 3741; https://doi.org/10.3390/foods13233741 - 22 Nov 2024
Viewed by 1411
Abstract
Cultured meat technology is a form of cellular agriculture where meat is produced from animal cells grown in a lab, instead of raising and slaughtering animals. This technology relies heavily on fetal bovine serum (FBS) in cell media; hence, production is costly and [...] Read more.
Cultured meat technology is a form of cellular agriculture where meat is produced from animal cells grown in a lab, instead of raising and slaughtering animals. This technology relies heavily on fetal bovine serum (FBS) in cell media; hence, production is costly and contributes significantly to ammonia and greenhouse gas emissions. Achieving the successful commercialization of cell-cultured food requires the critical resolution of manufacturing cost and safety concerns. Hence, our research efforts are focused on identifying commercially viable and ecologically sustainable alternatives to FBS. In this study, we evaluated the potential of twenty-six water-based algal and cyanobacterial extracts to stimulate cell growth for meat cultivation under 90% reduced serum conditions. The extracts were compared in viability, proliferation, and Trypan blue exclusion assays. In the first screening phase, the extracts were evaluated in a ZEM2S (zebrafish) cell culture in a 1% FBS regimen. Based on their ability to exhibit protein tolerance or promote cell proliferation, ten extracts were selected and further assayed in a QM7 cell culture. The QM7 cell line (myoblasts from Japanese quail) is highly relevant for meat cultivation because of its ability to differentiate into muscle fibers. Extracts derived from two microalgae species, Arthrospira platensis (Spirulina) and Dunaliella tertiolecta, demonstrated the highest tolerance in cell culture, above 10 μg/mL (expressed as total protein concentration). Tolerance at a 100 μg/mL concentration was demonstrated exclusively using an extract of blue spirulina (commercially purified Spirulina), which supported cell growth through multiple passages. Full article
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<p>We screened 26 algal/cyanobacterial extracts to determine their impact on the viability of ZEM2S cells. Cell viability was estimated in the Alamar Blue (resazurin) assay after 72 h of treatment and normalized to the non-treated/control cells, which were at 100%. Extracts were tested at 0.1, 1, 10, and 50 µg of protein per mL and, where possible, at 100 µg/mL. Each extract was tested at least twice; the graph presents the mean and SEM values of triplicate runs from one representative experiment. The legend contains the full names of the species, color-coded to match the columns and organized according to their phylum affiliation (for strain taxonomy, we used AlgaeBase: <a href="https://www.algaebase.org" target="_blank">https://www.algaebase.org</a>, accessed on 16 September 2024). Extracts selected for the second round of screening are shown in bold letters.</p>
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<p>Cytotoxicity of ten selected algal/cyanobacterial extracts on ZEM2S cells was measured by the neutral red uptake assay after 72 h of treatment and normalized to the non-treated/control cells, which was at 100%. Each extract was tested at least twice; mean and SEM values of triplicate runs from one representative experiment are presented on the graph. The legend contains the full names of the species, color-coded to match the columns. Statistically significant differences between the control and the treatment groups are labeled with * (<span class="html-italic">p</span> ≤ 0.05), ** (<span class="html-italic">p</span> ≤ 0.01), or **** (<span class="html-italic">p</span> ≤ 0.0001).</p>
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<p>The effect of the selected extracts on ZEM2S proliferation. Cell proliferation was assessed by measuring DNA synthesis in a BrdU incorporation assay at 48 h (<b>A</b>) and by counting cells in a Trypan blue exclusion assay at 72 h (<b>B</b>). Each extract was tested at least twice; mean and SEM values of triplicate runs from one representative experiment are presented on the graph. The legend contains the full names of the species, color-coded to match the columns. Statistically significant differences between the control and the treatment groups are labeled with * (<span class="html-italic">p</span> ≤ 0.05), ** (<span class="html-italic">p</span> ≤ 0.01), *** (<span class="html-italic">p</span> ≤ 0.001), or **** (<span class="html-italic">p</span> ≤ 0.0001).</p>
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<p>The effect of the selected extracts on QM7 cells. Cytotoxicity of five selected algal/cyanobacterial extracts was measured by the MTT assay (<b>A</b>) and Alamar Blue assay (<b>B</b>) after 72 h of treatment and normalized to the non-treated/control cells, which were at 100%. Cell proliferation was assessed by measuring DNA synthesis in a BrdU incorporation assay at 24–48 h (<b>C</b>). Each extract was tested at least twice; the graph presents the mean and SEM values of triplicate runs from one representative experiment. The legend contains the full names of the species, color-coded to match the columns. Statistically significant differences between the control and the treatment groups are labeled with * (<span class="html-italic">p</span> ≤ 0.05).</p>
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<p>Maintenance of ZEM2S and QM7 cells in the presence of blue spirulina (BS) and 1% FBS. Before the experiment, cells were routinely maintained in media supplemented with 10% FBS and passaged every three days. Upon seeding in 1% FBS, with or without 10 or 100 µg/mL of BS, the cell morphology and confluency were determined by image-based analysis. Cells were passaged, counted with Trypan blue (1–4 passages), re-seeded, and kept in culture for up to 32 days (1–6 passages). The multiplication factor was determined by dividing the number of living cells at the time of passage by the number of living cells at the time of seeding. (<b>A</b>) Multiplication factor for ZEM2S cells at 1–4 passages in BS-supplemented media; (<b>B</b>) confluency of ZEM2S cells at 0–32 days; (<b>C</b>) morphology of ZEM2S cells at passage 6; (<b>D</b>) multiplication factor for QM7 cells at 1–3 passages in BS-supplemented media; (<b>E</b>) confluency of QM7 cells at 0–32 days; (<b>F</b>) morphology of QM7 cells at passages 6 and 4. Statistically significant differences between the control and the treatment groups are labeled with * (<span class="html-italic">p</span> ≤ 0.05), ** (<span class="html-italic">p</span> ≤ 0.01), *** (<span class="html-italic">p</span> ≤ 0.001), or **** (<span class="html-italic">p</span> ≤ 0.0001).</p>
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16 pages, 1337 KiB  
Article
Embryonic Leucine Promotes Early Postnatal Growth via mTOR Signalling in Japanese Quails
by Sawadi F. Ndunguru, Gebrehaweria K. Reda, Brigitta Csernus, Renáta Knop, James K. Lugata, Csaba Szabó, Ádám Z. Lendvai and Levente Czeglédi
Animals 2024, 14(17), 2596; https://doi.org/10.3390/ani14172596 - 6 Sep 2024
Viewed by 1029
Abstract
Nutritional cues during embryonic development can alter developmental trajectories and affect postnatal growth. However, the specific mechanisms by which nutrients influence avian growth remain largely unknown. Amino acids can directly interact with the nutrient-sensing pathways, such as the insulin-like growth factor 1 (IGF-1)/mechanistic [...] Read more.
Nutritional cues during embryonic development can alter developmental trajectories and affect postnatal growth. However, the specific mechanisms by which nutrients influence avian growth remain largely unknown. Amino acids can directly interact with the nutrient-sensing pathways, such as the insulin-like growth factor 1 (IGF-1)/mechanistic target of rapamycin (mTOR) pathways, which are known to regulate growth. We examined the effects of embryonic leucine on gene expression and phenotypic growth in Japanese quails by injecting 2.5 mg leucine or saline (control) into Japanese quail eggs on the tenth day of incubation and incubating them under standard conditions. The treatment groups had similar hatching success and size at hatching. However, between 3 and 7 days post-hatching, quails treated with embryonic leucine showed increased growth in body mass and wing, tarsus, head, and intestinal lengths, lasting up to 21 days. The hepatic expression of IGF1, IGF1R, mTOR, and RPS6K1 was upregulated in leucine-treated quails, while the expression of FOXO1 remained unaffected. In conclusion, a subtle increase in embryonic leucine may induce developmental programming effects in Japanese quail by interacting with the IGF-1/mTOR nutrient-sensing pathway to promote growth. This study highlights the role of embryonic amino acids as crucial nutrients for enhancing growth. It provides valuable insight into nutrient intervention strategies during embryonic development to potentially improve poultry growth performance. Full article
(This article belongs to the Special Issue Amino Acid Nutrition in Poultry)
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<p>Leucine injection into Japanese quail eggs increased body mass (<b>A</b>) and head (<b>B</b>), tarsus (<b>C</b>) and wing (<b>D</b>) lengths in chicks post-hatch (see <a href="#app1-animals-14-02596" class="html-app">Table S1. Supplementary Materials</a> for detailed sample size). Asterisks indicate a significant difference between the treatment groups (<span class="html-italic">p</span> &lt; 0.05), and error bars indicate mean ± SE.</p>
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<p>Leucine injection into Japanese quail eggs increased postnatal intestinal length in 21-day-old chicks. Numbers in the bars indicate sample size (n). The asterisk indicates a significant difference between the treatment groups at <span class="html-italic">p</span> &lt; 0.05, and error bars indicate mean ± SE. Numbers 1 and 21 above the bars indicate the age of chicks in days post-hatch.</p>
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<p>Bar plots present mRNA expression relative to the reference gene. (<b>A</b>) <span class="html-italic">IGF1</span>, (<b>B</b>) <span class="html-italic">IGF1R</span>, (<b>C</b>) <span class="html-italic">mTOR</span>, (<b>D</b>) <span class="html-italic">RPS6K1</span>, (<b>E</b>) <span class="html-italic">FOXO1</span> expression, light grey bars refers to control group, dark grey bars refers to leucine treatment. Numbers in the bars indicate sample size (n). The numbers at the top denote the age of the Japanese quail chicks at which the samples were collected: (1) day-old chicks and (21) 21-day-old chicks. Asterisks indicate significant differences between the treatment groups (<span class="html-italic">p</span> &lt; 0.05), and error bars indicate mean ± SE.</p>
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10 pages, 666 KiB  
Article
Ileal Digestible and Metabolizable Energy of Corn, Wheat, and Barley in Growing Japanese Quail
by Sousan Khanipour, Mahmoud Ghazaghi, Mohammad Reza Abdollahi and Mehran Mehri
Poultry 2024, 3(3), 190-199; https://doi.org/10.3390/poultry3030015 - 24 Jun 2024
Viewed by 1806
Abstract
This study aimed to determine the ileal digestible energy (IDE), apparent metabolizable energy (AME), and nitrogen-corrected AME (AMEn) of three typical cereals for quail chicks with two age periods (day 15–21 and 22–28). The experimental diets comprised a corn–soybean meal reference diet (RD), [...] Read more.
This study aimed to determine the ileal digestible energy (IDE), apparent metabolizable energy (AME), and nitrogen-corrected AME (AMEn) of three typical cereals for quail chicks with two age periods (day 15–21 and 22–28). The experimental diets comprised a corn–soybean meal reference diet (RD), and three test diets (TD) that were fed to quail chicks in a completely randomized design with five replicates per diet and 15 birds each. The TD comprised corn, wheat, and barley that partly replaced the RD at 300 g/kg (70% reference diet + 30% test ingredient). Age did not influence the IDE, AME, and AMEn values, while the effect of ingredient type was highly significant on the energy estimates (p < 0.001). The IDE values of corn, wheat, and barley were estimated as 2924, 3440, and 3184 kcal/kg, respectively. The AME values of corn, wheat, and barley were 3519, 2979, and 2710 kcal/kg, respectively. The estimated AMEn values of corn, wheat, and barley were 3255, 2676, and 2281 kcal/kg, respectively. These findings are crucial for optimizing diet formulations to support quail growth and performance effectively, as they provide valuable insights into the energy content of different cereals for quail production. Notably, the high IDE and AME values of wheat suggest its potential as a valuable energy source for quail diets. Understanding these values can aid in formulating diets that meet the energy requirements of quail chicks, leading to improved growth rates, feed efficiency, and overall productivity in quail production systems. Full article
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<p>Comparison of the feed intake in quail chicks in the third (3rd) and fourth (4th) weeks of age.</p>
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<p>One sample <span class="html-italic">t</span>-test of the determined apparent metabolizable energy corrected for zero nitrogen (AMEn) of corn, wheat, and barley in quail chicks with the AMEn values reported in broiler chickens.</p>
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20 pages, 4942 KiB  
Article
Mitogenomic Insights into the Evolution, Divergence Time, and Ancestral Ranges of Coturnix Quails
by Prateek Dey, Swapna Devi Ray, Venkata Hanumat Sastry Kochiganti, Budhan S. Pukazhenthi, Klaus-Peter Koepfli and Ram Pratap Singh
Genes 2024, 15(6), 742; https://doi.org/10.3390/genes15060742 - 5 Jun 2024
Cited by 3 | Viewed by 1778
Abstract
The Old-World quails, Coturnix coturnix (common quail) and Coturnix japonica (Japanese quail), are morphologically similar yet occupy distinct geographic ranges. This study aimed to elucidate their evolutionary trajectory and ancestral distribution patterns through a thorough analysis of their mitochondrial genomes. Mitogenomic analysis revealed [...] Read more.
The Old-World quails, Coturnix coturnix (common quail) and Coturnix japonica (Japanese quail), are morphologically similar yet occupy distinct geographic ranges. This study aimed to elucidate their evolutionary trajectory and ancestral distribution patterns through a thorough analysis of their mitochondrial genomes. Mitogenomic analysis revealed high structural conservation, identical translational mechanisms, and similar evolutionary pressures in both species. Selection analysis revealed significant evidence of positive selection across the Coturnix lineage for the nad4 gene tree owing to environmental changes and acclimatization requirements during its evolutionary history. Divergence time estimations imply that diversification among Coturnix species occurred in the mid-Miocene (13.89 Ma), and their current distributions were primarily shaped by dispersal rather than global vicariance events. Phylogenetic analysis indicates a close relationship between C. coturnix and C. japonica, with divergence estimated at 2.25 Ma during the Pleistocene epoch. Ancestral range reconstructions indicate that the ancestors of the Coturnix clade were distributed over the Oriental region. C. coturnix subsequently dispersed to Eurasia and Africa, and C. japonica to eastern Asia. We hypothesize that the current geographic distributions of C. coturnix and C. japonica result from their unique dispersal strategies, developed to evade interspecific territoriality and influenced by the Tibetan Plateau’s geographic constraints. This study advances our understanding of the biogeographic and evolutionary processes leading to the diversification of C. coturnix and C. japonica, laying important groundwork for further research on this genus. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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<p>Circular schematic map of the mitogenomes of <span class="html-italic">C. coturnix</span> and <span class="html-italic">C. japonica</span>. Genes are represented with different color blocks, with PCGs in orange, rRNAs in blue, tRNAs in magenta, and the control region in green. Black sliding windows correspond to the GC content, and purple sliding windows correspond to the value of the GC skew. Letters in parentheses for tRNA tags indicate anticodons. The arrow direction at the end of protein-coding genes indicates transcription on the plus (right) or minus (left) strand.</p>
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<p>(<b>A</b>) <span class="html-italic">dN/dS</span> estimates for 13 protein-coding genes of <span class="html-italic">C. coturnix</span> and <span class="html-italic">C. japonica</span> mitogenomes sequenced in this study. The X-axis shows the gene name, and the Y-axis shows the units of the <span class="html-italic">dN/dS</span> ratio. (<b>B</b>) Maximum likelihood tree constructed using the <span class="html-italic">nad4</span> gene of selected mitogenomes used in this study. EasyCodeML, aBSREL, and BUSTED algorithms detected positive selection across the highlighted tree branch that leads to the <span class="html-italic">Coturnix</span> lineage (boldly marked in yellow ochre and asterisked).</p>
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<p>Topology of maximum likelihood (ML) and Bayesian inference (BI) trees generated for the 71 Phasianidae mitogenomes used in this study. <span class="html-italic">C. livia</span> is used as the out-group to root the trees. The numbers at each node represent bootstrap values for ML analyses (0–100%) and posterior probabilities for BI analyses (0–1). Branches of <span class="html-italic">Coturnix</span> clade members are in blue and highlighted in the dotted dark blue box. The <span class="html-italic">C. coturnix</span> and <span class="html-italic">C. japonica</span> mitogenomes sequenced in this study are highlighted in red text.</p>
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<p>BEAST maximum clade credibility tree showing divergence time estimates among Phasianidae species based on four fossil calibrations and rooted with the out-group, <span class="html-italic">C. livia</span>. Posterior probabilities are indicated only at nodes for values &lt; 1.0 in light blue. Mean divergence time estimates are shown in black next to the nodes, and purple bars show the lower and upper bounds of the 95% highest posterior density (95% HPD) interval for the time estimates. The branches of the <span class="html-italic">Coturnix</span> clade members are in red and highlighted in a dotted red box. The crimson asterisk indicates the nodes supported by fossil calibrations.</p>
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<p>The ancestral biogeographic area reconstruction estimated using the BayArea-like + J model generated in BioGeoBEARS. The BEAST maximum clade credibility tree was pruned to contain seven <span class="html-italic">Coturnix</span> and one <span class="html-italic">Alectoris</span> species (as the out-group). The estimated ancestral areas are colored according to the four zoogeographic zones: (i) A (Oceanian and Australian), (ii) B (Oriental), (iii) C (Palearctic and Sino-Japanese), and (iv) D (Saharo-Arabian, Afro-Tropical, and Madagascan). The color palette on the nodes of the chronogram shows the combination of most likely ancestral areas estimated by BioGeoBEARS. The coding scheme for each geographic area for each species, present or absent, is shown on the right of the tree. Insert: biogeographic areas used in the ancestral area reconstruction.</p>
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15 pages, 1269 KiB  
Article
Assessing the Impacts of Different Levels of Nano-Selenium on Growth Performance, Serum Metabolites, and Gene Expression in Heat-Stressed Growing Quails
by Rania Mahmoud, Basma Salama, Fatmah A. Safhi, Ioan Pet, Elena Pet and Ahmed Ateya
Vet. Sci. 2024, 11(6), 228; https://doi.org/10.3390/vetsci11060228 - 21 May 2024
Cited by 2 | Viewed by 1546
Abstract
Nano-minerals are employed to enhance mineral bioavailability thus promoting the growth and well-being of animals. In recent times, nano-selenium (nano-Se) has garnered significant attention within the scientific community owing to its potential advantages in the context of poultry. This study was conducted to [...] Read more.
Nano-minerals are employed to enhance mineral bioavailability thus promoting the growth and well-being of animals. In recent times, nano-selenium (nano-Se) has garnered significant attention within the scientific community owing to its potential advantages in the context of poultry. This study was conducted to explore the impact of using variable levels of nano-Se on the growth performance, carcass characteristics, serum constituents, and gene expression in growing Japanese quails under both thermoneutral and heat stress conditions. A randomized experimental design was used in a 2 × 3 factorial, with 2 environmental conditions (thermoneutral and heat stress) and 3 nano-Se levels (0, 0.2, and 0.5 mg/kg of diet. The findings revealed that heat stress negatively affected the growth and feed utilization of quails; indicated by the poor BWG and FCR. Additionally, oxidative stress was aggravated under heat stress condition; indicated by increased lipids peroxidation and decreased antioxidant enzymes activities. The addition of nano-Se, especially at the level of 0.2 mg/kg of diet, significantly improved the performance of heat stressed quails and restored blood oxidative status. The expression profile of inflammatory and antioxidant markers was modulated by heat stress and/or 0.2 and 0.5 nano-Se in conjunction with environmental temperature in quail groups. In comparison to the control group, the heat stress-exposed quails’ expression profiles of IL-2, IL-4, IL-6, and IL-8 showed a notable up-regulation. Significantly lower levels of the genes for IL-2, IL-4, IL-6, and IL-8 and higher levels of the genes for SOD and GPX as compared to the heat stress group demonstrated the ameliorative impact of 0.2 nano-Se. The expression profiles of IL-2, IL-4, IL-6, and IL-8 are dramatically elevated in quails exposed to 0.5 nano-Se when compared to the control group. SOD and GPX markers, on the other hand, were markedly down-regulated. It was concluded that nano-Se by low level in heat stressed growing quails provides the greatest performance and its supplementation can be considered as a protective management practice in Japanese quail diets to reduce the negative impact of heat stress. Full article
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<p>The effect of different nano-Se levels and environmental temperature on quail’s blood lipid peroxidation and antioxidant enzymes levels. MDA (<b>A</b>), SOD (<b>B</b>), GPx (<b>C</b>) and GSH (<b>D</b>) levels were measured in blood samples and data are presented as mean ± SEM. Different upper-case letters (A–C) show significant differences (<span class="html-italic">p</span> &lt; 0.05) among different concentrations of nano-Se supplementation under heat stress condition. Different lower-case letters (a–c) exhibit significant differences (<span class="html-italic">p</span> &lt; 0.05) among different concentrations of nano-Se supplementation under thermoneutral condition. Asterisk (*) represents statistical difference between thermoneutral and heat stress subjected groups, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>Muscles Se levels under the effect of different nano-Se supplementations and different environmental temperature. Se was measured in muscle tissue samples by ICP-MS. Data are presented as mean ± SEM. Different upper-case letters (A–C) show significant differences (<span class="html-italic">p</span> &lt; 0.05) among different concentrations of nano-Se supplementation under heat stress condition. Different lower-case letters (a–c) exhibit significant differences (<span class="html-italic">p</span> &lt; 0.05) among different concentrations of nano-Se supplementation under thermoneutral condition. Asterisk (*) represents statistical difference between thermoneutral and heat stress subjected groups, * <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Effect of different nano-Se levels and environmental temperature on gene expression of inflammatory and antioxidant markers. Data are presented as mean ± SEM. Different upper-case letters (A–C) show significant differences (<span class="html-italic">p</span> &lt; 0.05) among different concentrations of nano-Se supplementation under heat stress condition. Different lower-case letters (a–c) exhibit significant differences (<span class="html-italic">p</span> &lt; 0.05) among different concentrations of nano-Se supplementation under thermoneutral condition. Asterisk (**) represents statistical difference between thermoneutral and heat stress subjected groups, ** <span class="html-italic">p</span> &lt; 0.01.</p>
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21 pages, 5247 KiB  
Article
Antihypertensive Potential of Japanese Quail (Couturnix Couturnix Japonica) Egg Yolk Oil (QEYO) in Sprague Dawley Rats
by Muhammad Sani Ismaila, Sherifat Olayemi Balogun-Raji, Fahad Hamza, Usman Bello Sadiya, Buhari Salisu, Mohammed Umar, Ishaka Aminu and Kegan Romelle Jones
Biology 2024, 13(4), 270; https://doi.org/10.3390/biology13040270 - 18 Apr 2024
Viewed by 1792
Abstract
Oils from animal sources have been used for centuries in the management of diseases. This research was conducted to screen the ex vivo and in vivo toxicity of quail egg yolk oil (QEYO) extracts and assess their effects on the management of hypertension [...] Read more.
Oils from animal sources have been used for centuries in the management of diseases. This research was conducted to screen the ex vivo and in vivo toxicity of quail egg yolk oil (QEYO) extracts and assess their effects on the management of hypertension in rats. QEYO was extracted using gentle heating (GH) and n-hexane (NHN). The extracts were subjected to toxicity testing using the hen’s egg test on chorioallantoic membrane (HET-CAM) and bovine corneal histology test. Acute and sub-chronic toxicity (28 days) were evaluated in rats. Hypertension was induced in rats by administering 80 mg/kg of Nω-L-Arginine Methyl Ester (L-NAME) per day for 28 days. Treatments commenced on the 14th day; Nifedipine at 30 mg/kg and 1 mL of distilled water were administered as positive and negative controls. Blood pressure (BP), lipid profiles, and oxidative stress markers were quantified. No irritation was observed using the HET-CAM test in the egg treated with both extracts. Bovine corneal histology showed no lesions in all treated groups. No signs of toxicity were observed in either acute or sub-chronic toxicity studies. A significant reduction in blood pressure was observed in rats treated with the extracts (p < 0.05). Changes in total cholesterol (TC), triglycerides (TGs), low-density lipoproteins (LDLPs), and high-density lipoproteins (HDLPs) were not significant compared to the control (p > 0.05). Oxidative stress markers (SOD and CAT) increased significantly in the treated groups compared to the control, while the malondialdehyde levels decreased (p < 0.05). QEYO was safe in both ex vivo and in vivo studies and can be said to have the potential to lower blood pressure as well as cardio-protective effects in hypertensive rats. This research provides evidence based on which QEYO could be used safely as an adjuvant therapy in eye drops and cosmetics and can be considered an effective choice for preventing hypertension. Full article
(This article belongs to the Special Issue Pathophysiology of Hypertension and Related Diseases)
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<p>HET-CAM irritation. The arrows indicate observation areas on the egg CAMs. (<b>A</b>) Before treatment with 0.1 mL Na OH (negative control showing no sign of toxicity or coagulation on the blood vessels); (<b>B</b>) after treatment with 0.2 mL Na OH (positive control showing irritation and coagulation of blood on the vessels); (<b>C</b>) after treatment with 0.2 mL n-hexane quail egg yolk oil extract (showing no irritation or coagulation of blood); (<b>D</b>) after treatment with 0.2 mL gentle heating quail egg yolk oil (showing no toxicity to the blood vessels).</p>
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<p>Gross appearance of the bovine cornea after inoculation with the positive control and different concentrations of the test substance: (<b>A</b>) 0.1 mL normal saline (negative control); (<b>B</b>) 100% ethanol (positive control); (<b>C</b>) 0.1 mL (QEYO-GH); (<b>D</b>) 0.1 mL (QEYO-n-Hexane); (<b>E</b>) 0.2 mL (QEYO-GH); (<b>F</b>) 0.2 mL (QEYO-n-Hexane).</p>
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<p>Histopathological sections of bovine corneas showing the negative control (0.1 mL of 0.9%NaCl) and positive control (0.1 mL of 100% ethanol) as well as the QEYONH (0.1 and 0.2 mL)-treated and QEYOGH (0.1 and 0.2 mL)-treated groups. (<b>1a</b>) Corneal stratified squamous epithelium (white arrow), collagenous stroma (black arrows), and Descemet’s membrane (long black arrow); (<b>1b</b>) corneal stratified squamous epithelium (white arrow), collagenous stroma (blue arrows), and ciliary body (double black arrows). (<b>2a</b>) Corneal stratified squamous epithelium (white arrow), collagenous stroma (black arrows), and Descemet’s membrane (long black arrow); (<b>2b</b>) ciliary body (white arrows) and scleral stroma (double black arrows); (<b>2c</b>) retina (white arrow) and scleral stroma (double black arrows); (<b>2d</b>) optic nerve. (<b>3a</b>) Corneal stratified squamous epithelium (white arrow), collagenous stroma (double black arrows), and Descemet’s membrane (long arrow); (<b>3b</b>) corneal stratified squamous epithelium (white arrow), collagenous stroma (black arrows head), and ciliary body (double black arrows); (<b>3c</b>) retina (white arrow) and scleral stroma (double black arrows). (<b>4a</b>) Corneal stratified squamous epithelium (white arrow), collagenous stroma (double black arrows), and Descemet’s membrane (long arrow); (<b>4b</b>) corneal stratified squamous epithelium (white arrow), collagenous stroma (black arrows head), and ciliary body (double black arrows); (<b>4c</b>) retina (white arrow) and scleral stroma (double black arrows); (<b>4d</b>) the optic nerve. All the slides were stained using hematoxylin and eosin and viewed using light microscopy at a ×100 magnification.</p>
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<p>Histological sections of the liver, kidney, and brain. (<b>1a</b>) Liver: section show normal portal triad (arrow) and hepatocytes arranged in cords (short arrow); H&amp;E × 100. (<b>1b</b>) Kidney: section shows regular glomerulus (Long arrow) and renal tubules (short arrow); H&amp;E × 100. (<b>1c</b>) Brain: section shows regular neutrophils; H&amp;E × 100. (<b>2a</b>) Liver: section shows normal hepatic central vein (long arrow), hepatocytes arranged in cords (short arrow), and sinusoidal space (arrow head); H&amp;E × 100. (<b>2b</b>) Kidney: section shows regular glomerulus (long arrow) and renal tubules (short arrow); H&amp;E × 100. (<b>2c</b>) Brain: section show regular neutrophils; H&amp;E × 100. (<b>3a</b>) Liver: section show normal hepatic central vein (long arrow), hepatocytes arranged in cords (short arrow), and sinusoidal space (arrow head); H&amp;E × 100. (<b>3b</b>) Kidney: section shows regular glomerulus (long arrow) and renal tubules (short arrow); H&amp;E × 100. (<b>3c</b>) Brain: section show regular neutrophils; H&amp;E × 100.</p>
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<p>The mean SBP plot comparison from day 0 to day 28. Systolic blood pressure of rats treated with L-NAME (80 mg/kg) followed by treatment with different doses of quail egg yolk oil extracted using gentle heating (QEYOGH) and quail egg yolk oil extracted using <span class="html-italic">n</span>-hexane (QEYONH). Nifedipine (30 mg/kg/day) was used as a standard control drug. * Indicates a significant decrease in SBP in the groups treated with the higher doses of the extract and the positive control compared to the control non-treated group at 21 and 28 days (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Diastolic blood pressure of rats treated with L-NAME (80 mg/kg) followed by treatment with different doses of quail egg yolk oil extracted using gentle heating (QEYOGH) and quail egg yolk oil extracted using <span class="html-italic">n</span>-hexane (QEYONH). Nifedipine (30 mg/kg/day) was used as a standard control drug. * Indicates a significant decrease in DBP in the groups treated with the higher doses of the extract and the positive control compared to the control non-treated group at 21 and 28 days (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p><b>Plate 1</b>: Photomicrograph of the heart of a rat induced with hypertension using L-NAME (80 mg/kg/day) and treated with 1 mL/kg of distilled water showing loss of architecture of the heart and oedema. There is also narrowing of the coronary artery due to thickness around the arterial lumen, H&amp;E × 100; A = loss of architecture, B = narrowing of arterial lumen, C = arteriosclerosis. <b>Plate 2</b>: Photomicrograph of the heart of a rat induced with hypertension using L-NAME (80 mg/kg/day) and treated with nifedipine at 30 mg/kg/day showing normal architecture of the heart with normal myocytes and coronary artery of the heart, H&amp;E × 100; A = normal heart wall. <b>Plate 3</b>: Photomicrograph of the heart of a rat induced with hypertension using L-NAME (80 mg/kg/day) and administered QEONH at 300 mg/kg showing slight enlargement of the myocytes and coronary artery of the heart, H&amp;E × 100; A = slight enlargement of arterial wall. <b>Plate 4</b>: Photomicrograph of the heart of a rat induced with hypertension using L-NAME (80 mg/kg/day) and treated with quail egg yolk oil obtained by gentle hearting at 300 mg/kg showing normal architecture of the heart with normal myocytes and coronary artery, H&amp;E × 100; A = coronary arterial wall, B = arterial lumen.</p>
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17 pages, 4935 KiB  
Article
Genome-Wide Association Study Revealed Putative SNPs and Candidate Genes Associated with Growth and Meat Traits in Japanese Quail
by Natalia A. Volkova, Michael N. Romanov, Alexandra S. Abdelmanova, Polina V. Larionova, Nadezhda Yu. German, Anastasia N. Vetokh, Alexey V. Shakhin, Ludmila A. Volkova, Alexander A. Sermyagin, Dmitry V. Anshakov, Vladimir I. Fisinin, Darren K. Griffin, Johann Sölkner, Gottfried Brem, John C. McEwan, Rudiger Brauning and Natalia A. Zinovieva
Genes 2024, 15(3), 294; https://doi.org/10.3390/genes15030294 - 25 Feb 2024
Cited by 4 | Viewed by 1805
Abstract
The search for SNPs and candidate genes that determine the manifestation of major selected traits is one crucial objective for genomic selection aimed at increasing poultry production efficiency. Here, we report a genome-wide association study (GWAS) for traits characterizing meat performance in the [...] Read more.
The search for SNPs and candidate genes that determine the manifestation of major selected traits is one crucial objective for genomic selection aimed at increasing poultry production efficiency. Here, we report a genome-wide association study (GWAS) for traits characterizing meat performance in the domestic quail. A total of 146 males from an F2 reference population resulting from crossing a fast (Japanese) and a slow (Texas White) growing breed were examined. Using the genotyping-by-sequencing technique, genomic data were obtained for 115,743 SNPs (92,618 SNPs after quality control) that were employed in this GWAS. The results identified significant SNPs associated with the following traits at 8 weeks of age: body weight (nine SNPs), daily body weight gain (eight SNPs), dressed weight (33 SNPs), and weights of breast (18 SNPs), thigh (eight SNPs), and drumstick (three SNPs). Also, 12 SNPs and five candidate genes (GNAL, DNAJC6, LEPR, SPAG9, and SLC27A4) shared associations with three or more traits. These findings are consistent with the understanding of the genetic complexity of body weight-related traits in quail. The identified SNPs and genes can be used in effective quail breeding as molecular genetic markers for growth and meat characteristics for the purpose of genetic improvement. Full article
(This article belongs to the Special Issue Poultry Genetics and Genomics)
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Figure 1
<p>Assortment of quail products: wholesale commercial egg tray and a pack of seasoned smoked hard-boiled eggs (in the foreground); whole egg mayonnaise, quail egg nutrient noodles, and frozen quail carcasses (in the background). Credit: authors’ own photo.</p>
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<p>Quails of the F<sub>2</sub> reference population: (<b>A</b>) at 3 days of age and (<b>B</b>) at 7 weeks of age.</p>
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<p>Overview of the experimental design. The reference population consisting of 146 F<sub>2</sub> male quails was created and extensively phenotyped using reciprocal crosses between a slow-growing Japanese and a fast-growing Texas White breed. <span class="html-italic">n</span>, number of progenies (of both sexes) from each F<sub>2</sub> family.</p>
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<p>Principal component analysis (PCA) of the F<sub>2</sub> quail reference population based on GBS data. (<b>A</b>) PCA performed in the plane of the first (PC1, <span class="html-italic">X</span>-axis) and second (PC2, <span class="html-italic">Y</span>-axis) components. (<b>B</b>) PCA performed in the plane of the first (PC1, <span class="html-italic">X</span>-axis) and second (PC3, <span class="html-italic">Y</span>-axis) components. Individuals from different groups are indicated by different colors.</p>
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<p>Manhattan plots of the GWAS results for the studied meat production traits: (<b>A</b>) body weight at 1 day of age, (<b>B</b>) body weight at 56 days of age, (<b>C</b>) average daily body weight gain, (<b>D</b>) dressed carcass weight, (<b>E</b>) breast weight, (<b>F</b>) thigh weight, and (<b>G</b>) drumstick weight. Distribution of SNPs in quail chromosomes for single traits are shown relative to the thresholds for the genome-wide nominal significance level (−log<sub>10</sub> (<span class="html-italic">p</span>)) according to the estimated probability values of <span class="html-italic">p</span> &lt; 1.0 × 10<sup>−5</sup> (lower line) and <span class="html-italic">p</span> &lt; 5.4 × 10<sup>−7</sup> (upper line). Points are color-coded only to visualize chromosome separation.</p>
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<p>Manhattan plots of the GWAS results for the studied meat production traits: (<b>A</b>) body weight at 1 day of age, (<b>B</b>) body weight at 56 days of age, (<b>C</b>) average daily body weight gain, (<b>D</b>) dressed carcass weight, (<b>E</b>) breast weight, (<b>F</b>) thigh weight, and (<b>G</b>) drumstick weight. Distribution of SNPs in quail chromosomes for single traits are shown relative to the thresholds for the genome-wide nominal significance level (−log<sub>10</sub> (<span class="html-italic">p</span>)) according to the estimated probability values of <span class="html-italic">p</span> &lt; 1.0 × 10<sup>−5</sup> (lower line) and <span class="html-italic">p</span> &lt; 5.4 × 10<sup>−7</sup> (upper line). Points are color-coded only to visualize chromosome separation.</p>
Full article ">Figure 5 Cont.
<p>Manhattan plots of the GWAS results for the studied meat production traits: (<b>A</b>) body weight at 1 day of age, (<b>B</b>) body weight at 56 days of age, (<b>C</b>) average daily body weight gain, (<b>D</b>) dressed carcass weight, (<b>E</b>) breast weight, (<b>F</b>) thigh weight, and (<b>G</b>) drumstick weight. Distribution of SNPs in quail chromosomes for single traits are shown relative to the thresholds for the genome-wide nominal significance level (−log<sub>10</sub> (<span class="html-italic">p</span>)) according to the estimated probability values of <span class="html-italic">p</span> &lt; 1.0 × 10<sup>−5</sup> (lower line) and <span class="html-italic">p</span> &lt; 5.4 × 10<sup>−7</sup> (upper line). Points are color-coded only to visualize chromosome separation.</p>
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10 pages, 279 KiB  
Article
The Behavioral and Productive Characteristics of Japanese Quails (Coturnix japonica) Exposed to Different Monochromatic Lighting
by Sezgi Karal, Firdevs Korkmaz Turgud, Doğan Narinç and Ali Aygun
Animals 2024, 14(3), 482; https://doi.org/10.3390/ani14030482 - 1 Feb 2024
Viewed by 1545
Abstract
The purpose of this study is to examine the impact of monochromatic illuminations at wavelengths of white (400–700 nm), green (560 nm), yellow (580 nm), blue (480 nm), and red (660 nm) on the performance and behavioral traits of Japanese quails throughout their [...] Read more.
The purpose of this study is to examine the impact of monochromatic illuminations at wavelengths of white (400–700 nm), green (560 nm), yellow (580 nm), blue (480 nm), and red (660 nm) on the performance and behavioral traits of Japanese quails throughout their fattening period. A total of 300 quails in five lighting experimental groups were housed in their conventional rearing cages. Weekly live weights of quails were measured individually, developmental stability was determined, and behavior and fear tests were performed. The body weight averages of quails exposed to blue, green, and yellow light were determined to be greater than those exposed to white and red light (p < 0.05). In terms of the mature weight parameter and the weight of the inflection point of the Gompertz growth model, the averages of the quails exposed to green and blue monochromatic lighting were higher (p < 0.05). The most negative findings on aggressive behavior were observed in birds exposed to monochromatic yellow and red light. Although the body weight of quails exposed to yellow monochromatic lighting was similar to that of quails exposed to green and blue monochromatic lighting, green and blue monochromatic lighting produced the best results in terms of growth, behavior, and developmental stability characteristics. Consequently, it is believed that consistently using green or blue monochromatic lighting programs when raising Japanese quails may provide economic advantages to the producers. Full article
(This article belongs to the Collection Current Advances in Poultry Research)
16 pages, 2520 KiB  
Article
STC2 Inhibits Hepatic Lipid Synthesis and Correlates with Intramuscular Fatty Acid Composition, Body Weight and Carcass Traits in Chickens
by Yuzhu Cao, Qihui Jia, Yuxin Xing, Chenglin Ma, Hongbo Guan, Weihua Tian, Xiangtao Kang, Yadong Tian, Xiaojun Liu and Hong Li
Animals 2024, 14(3), 383; https://doi.org/10.3390/ani14030383 - 25 Jan 2024
Cited by 1 | Viewed by 1434
Abstract
Stanniocalcin 2 (STC2) is a secreted glycoprotein involved in multiple biological processes. To systemically study the biological role of STC2 in chickens, phylogenetic tree analysis and conservation analysis were conducted. Association analysis between variations in the STC2 gene and the economic traits of [...] Read more.
Stanniocalcin 2 (STC2) is a secreted glycoprotein involved in multiple biological processes. To systemically study the biological role of STC2 in chickens, phylogenetic tree analysis and conservation analysis were conducted. Association analysis between variations in the STC2 gene and the economic traits of Gushi-Anka F2 was conducted. The tissue expression patterns of STC2 expression in different chicken tissues and liver at different stages were detected. The biological role of STC2 in chicken liver was investigated through overexpression and interfering methods in the LMH cell line. Correlation analyses between STC2 expression and lipid components were conducted. (1) The phylogenetic tree displayed that chicken STC2 is most closely related with Japanese quail and most distantly related with Xenopus tropicalis. STC2 has the same identical conserved motifs as other species. (2) rs9949205 (T > C) found in STC2 intron was highly significantly correlated with chicken body weight at 0, 2, 4, 6, 8, 10 and 12 weeks (p < 0.01). Extremely significant correlations of rs9949205 with semi-evisceration weight (SEW), evisceration weight (EW), breast muscle weight (BMW), leg muscle weight (LMW), liver weight and abdominal fat weight (AFW) were revealed (p < 0.01). Significant associations between rs9949205 and abdominal fat percentage, liver weight rate, breast muscle weight rate and leg muscle weight rate were also found (p < 0.05). Individuals with TT or TC genotypes had significantly lower abdominal fat percentage and liver weight rate compared to those with the CC genotype, while their body weight and other carcass traits were higher. (3) STC2 showed a high expression level in chicken liver tissue, which significantly increased with the progression of age (p < 0.05). STC2 was observed to inhibit the content of lipid droplets, triglycerides (TG) and cholesterol (TC), as well the expression level of genes related to lipid metabolism in LMH cells. (4) Correlation analysis showed that the STC2 gene was significantly correlated with 176 lipids in the breast muscle (p < 0.05) and mainly enriched in omega-3 and omega-6 unsaturated fatty acids. In conclusion, the STC2 gene in chicken might potentially play a crucial role in chicken growth and development, as well as liver lipid metabolism and muscle lipid deposition. This study provides a scientific foundation for further investigation into the regulatory mechanism of the STC2 gene on lipid metabolism and deposition in chicken liver. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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<p>Phylogenetic tree and conservation analysis of STC2 in different species. (<b>A</b>) Phylogenetic analysis of STC2 amino acid sequences for species including chicken (<span class="html-italic">Gallus gallus</span>), Japanese quail (<span class="html-italic">Coturnix japonica</span>), green sea turtle (<span class="html-italic">Chelonia mydas</span>), human (<span class="html-italic">Homo sapiens</span>), Norway rat (<span class="html-italic">Rattus norvegicus</span>), tropical (<span class="html-italic">Xenopus tropicalis</span>). Using MEGA10.0 software, the amino acid sequences of STC2 from six species were selected and a rootless neighbor-joining phylogenetic tree was constructed, and the bootstrap test was set to repeat 2000 times. (<b>B</b>) Distribution of STC2 conserved motifs in different species. Different colored boxes indicate different conserved motif sequences.</p>
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<p>Expression characteristics of <span class="html-italic">STC2</span>. (<b>A</b>) Expression pattern of <span class="html-italic">STC2</span> in different tissues of Lushi hens at 30 weeks old (<span class="html-italic">n</span> = 8). (<b>B</b>) The spatio-temporal expression of <span class="html-italic">STC2</span> in liver tissue of Lushi chickens at different stages (<span class="html-italic">n</span> = 8). w means week. <span class="html-italic">n</span> ≥ 6 for each group. Each dot represents an individual. The relative expression of genes was normalized to <span class="html-italic">GAPDH</span>. Different letters mean significant difference (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effects of overexpression of <span class="html-italic">STC2</span> gene on lipid metabolism in LMH cells. (<b>A</b>) The overexpression efficiency of <span class="html-italic">STC2</span>. (<b>B</b>) Contents of TC and TG in LMH cells. (<b>C</b>) Lipid accumulation was evaluated with Oil Red O staining and quantified by absorbance value of the extracted Oil Red O dye. Each dot represents a repetition (<span class="html-italic">n</span> ≥ 4). * indicates <span class="html-italic">p</span> &lt; 0.05; ** indicates <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>Effect of overexpression of <span class="html-italic">STC2</span> gene on genes related to lipid metabolism in LMH cells. (<b>A</b>) Relative expression levels of TC-synthesis-related genes. (<b>B</b>) Relative expression levels of fatty-acid-synthesis-related genes. (<b>C</b>) Relative expression levels of TG-synthesis-related genes. (<b>D</b>) Relative expression levels of lipid-transporter-related genes. Each dot represents a repetition (<span class="html-italic">n</span> ≥ 4). The mRNA levels of genes were normalized to <span class="html-italic">GAPDH</span>. * indicates <span class="html-italic">p</span> &lt; 0.05; ** indicates <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>Effects of interfering with <span class="html-italic">STC2</span> gene on lipid metabolism in LMH cells. (<b>A</b>) Interference with <span class="html-italic">STC2</span> gene efficiency assay. (<b>B</b>) Contents of TC and TG in LMH cells. (<b>C</b>) Lipid accumulation was evaluated with Oil Red O staining and quantified by absorbance value of the extracted Oil Red O dye. Each dot represents a repetition (<span class="html-italic">n</span> ≥ 4). * indicates <span class="html-italic">p</span> &lt; 0.05; ** indicates <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>Effects of interfering with <span class="html-italic">STC2</span> gene on genes related to lipid metabolism in LMH cells. (<b>A</b>) Relative expression levels of TC-synthesis-related genes. (<b>B</b>) Relative expression levels of fatty-acid-synthesis-related genes. (<b>C</b>) Relative expression levels of TG-synthesis-related genes. (<b>D</b>) Relative expression levels of lipid-transporter-related genes. Each dot represents a repetition (<span class="html-italic">n</span> ≥ 4). The mRNA levels of genes were normalized to <span class="html-italic">GAPDH</span>. * indicates <span class="html-italic">p</span> &lt; 0.05; ** indicates <span class="html-italic">p</span> &lt; 0.01.</p>
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<p><span class="html-italic">STC2</span> expression contributes to the long-chain unsaturated glycerophospholipids deposition in intramuscular fat of Gushi chicken. (<b>A</b>) Transcriptomic data of the <span class="html-italic">STC2</span> gene in the 43-week high and low intramuscular adiposity group. G43wHM (<span class="html-italic">n</span> = 8) indicates high intramuscular fat group; G43wLM (<span class="html-italic">n</span> = 8) indicates low intramuscular fat group. (<b>B</b>) Lipid molecules in intramuscular fat significantly correlated with <span class="html-italic">STC2</span> expression in pectoralis of Gushi chicken. (<b>C</b>) Map of 130 positively correlated glycerophospholipid lipid molecules. (<b>D</b>) Proportion of different types of fatty acids at sn-1 and sn-2 positions of the positively correlated glycerophospholipid molecules. SFA (<span class="html-italic">n</span> = 0) indicates saturated fatty acids, MUFA (<span class="html-italic">n</span> = 1) indicates monounsaturated fatty acids and PUFA (<span class="html-italic">n</span> ≥ 2) indicates polyunsaturated fatty acids. (<b>E</b>) Proportional stacking of different types of MUFAs at sn-1 and sn-2 positions in (<b>D</b>). * indicates <span class="html-italic">p</span> &lt; 0.05.</p>
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17 pages, 5257 KiB  
Article
Metabolic Rate and Egg Production in Japanese Quails Can Be Predicted by Assessing Growth Parameters of Laying Hens
by Valeriy G. Narushin, Natalia A. Volkova, Anastasia N. Vetokh, Alan Yu. Dzhagaev, Ludmila A. Volkova, Darren K. Griffin, Michael N. Romanov and Natalia A. Zinovieva
Animals 2024, 14(2), 258; https://doi.org/10.3390/ani14020258 - 14 Jan 2024
Cited by 3 | Viewed by 1403
Abstract
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 [...] Read more.
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. Full article
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<p>Two related <span class="html-italic">Coturnix</span> species, the wild common (<span class="html-italic">C. coturnix</span>; (<b>a</b>)) and the domestic Japanese (<span class="html-italic">C. japonica</span>; (<b>b</b>)) quails, with a schematic representation of the ellipse (red line) based on the measurements of the bird’s body length and chest circumference. Image sources: (<b>a</b>) <a href="https://commons.wikimedia.org/wiki/File:Coturnix_coturnix_1873.jpg" target="_blank">https://commons.wikimedia.org/wiki/File:Coturnix_coturnix_1873.jpg</a> (accessed on 12 January 2024), John Gerrard Keulemans (1842–1912), Onze vogels in huis en tuin (1873), Creative Commons Public Domain Mark 1.0 license (CC-PD-Mark) and (<b>b</b>) <a href="https://commons.wikimedia.org/wiki/File:202205_Japanese_quail.svg" target="_blank">https://commons.wikimedia.org/wiki/File:202205_Japanese_quail.svg</a> (accessed on 12 January 2024), DataBase Center for Life Science (DBCLS), <a href="https://doi.org/10.7875/togopic.2022.186" target="_blank">https://doi.org/10.7875/togopic.2022.186</a> (accessed on 12 January 2024), Creative Commons Attribution 4.0 International license (CC-BY-4.0).</p>
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<p>Linear changes in growth parameters among eight quail breeds: (<b>a</b>) body weight (<span class="html-italic">B</span>), (<b>b</b>) body volume (<span class="html-italic">V</span>), and (<b>c</b>) body surface area (<span class="html-italic">S</span>). Breeds: TEW, Texas White; EST, Estonian; PHA, Pharaoh; MAG, Manchurian Golden; ENW, English White; JAP, Japanese; TUX, Tuxedo; and ENB, English Black.</p>
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<p>Changes in the <span class="html-italic">S</span>/<span class="html-italic">V</span> ratio among eight quail breeds. Breeds: TEW, Texas White; EST, Estonian; PHA, Pharaoh; MAG, Manchurian Golden; ENW, English White; JAP, Japanese; TUX, Tuxedo; and ENB, English Black.</p>
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<p>Relationship between body surface area (<span class="html-italic">S</span>) and volume (<span class="html-italic">V</span>) among eight quail breeds. Breeds: TEW, Texas White; EST, Estonian; PHA, Pharaoh; MAG, Manchurian Golden; ENW, English White; JAP, Japanese; TUX, Tuxedo; and ENB, English Black.</p>
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<p>Dependencies for performance traits, i.e., (<b>a</b>) number of eggs laid (<span class="html-italic">N</span>); (<b>b</b>) average egg weight (<span class="html-italic">W</span>); and (<b>c</b>) total egg mass (<span class="html-italic">M</span>) for three months of the assessed period, respectively, relative to the body weight of quails (<span class="html-italic">B</span>), volume (<span class="html-italic">V</span>), and surface area of their body (<span class="html-italic">S</span>).</p>
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<p>Correlation between the number of eggs laid (<span class="html-italic">N</span>) over three months of the assessed period and the metabolic level of quails, expressed by the <span class="html-italic">S</span>/<span class="html-italic">V</span> ratio.</p>
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<p>Correlation of the metabolism level of quails, expressed by the <span class="html-italic">S</span>/<span class="html-italic">V</span> ratio, with the body weight of laying hens (<span class="html-italic">B</span>).</p>
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<p>Dependencies for performance traits, i.e., (<b>a</b>) number of eggs laid (<span class="html-italic">N</span>); (<b>b</b>) average egg weight (<span class="html-italic">W</span>); and (<b>c</b>) total egg mass (<span class="html-italic">M</span>), for three months of the assessed period relative to the quail metabolic index <span class="html-italic">B·S</span>/<span class="html-italic">V</span>.</p>
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<p>Graphical dependence of the surface area (<span class="html-italic">S</span>) of a quail on its body weight (<span class="html-italic">B</span>).</p>
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<p>Correlation dependencies of the average egg weight, W (<b>a</b>), and the total egg mass for three months, <span class="html-italic">M</span> (<b>b</b>), with the slopes of trend lines reflecting changes in weight (TAN<span class="html-italic">B</span>), volume (TAN<span class="html-italic">V</span>), and surface area of quail body (TAN<span class="html-italic">S</span>).</p>
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<p>Correlation dependence of the total egg mass (<span class="html-italic">M</span>) for three months with the slope angles of the trend lines reflecting changes in the quail metabolic index, TAN(<span class="html-italic">B·S</span>/<span class="html-italic">V</span>).</p>
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22 pages, 6388 KiB  
Article
Genotyping-by-Sequencing Strategy for Integrating Genomic Structure, Diversity and Performance of Various Japanese Quail (Coturnix japonica) Breeds
by Natalia A. Volkova, Michael N. Romanov, Alexandra S. Abdelmanova, Polina V. Larionova, Nadezhda Yu. German, Anastasia N. Vetokh, Alexey V. Shakhin, Ludmila A. Volkova, Dmitry V. Anshakov, Vladimir I. Fisinin, Valeriy G. Narushin, Darren K. Griffin, Johann Sölkner, Gottfried Brem, John C. McEwan, Rudiger Brauning and Natalia A. Zinovieva
Animals 2023, 13(22), 3439; https://doi.org/10.3390/ani13223439 - 7 Nov 2023
Cited by 4 | Viewed by 1555
Abstract
Traces of long-term artificial selection can be detected in genomes of domesticated birds via whole-genome screening using single-nucleotide polymorphism (SNP) markers. This study thus examined putative genomic regions under selection that are relevant to the development history, divergence and phylogeny among Japanese quails [...] Read more.
Traces of long-term artificial selection can be detected in genomes of domesticated birds via whole-genome screening using single-nucleotide polymorphism (SNP) markers. This study thus examined putative genomic regions under selection that are relevant to the development history, divergence and phylogeny among Japanese quails of various breeds and utility types. We sampled 99 birds from eight breeds (11% of the global gene pool) of egg (Japanese, English White, English Black, Tuxedo and Manchurian Golden), meat (Texas White and Pharaoh) and dual-purpose (Estonian) types. The genotyping-by-sequencing analysis was performed for the first time in domestic quails, providing 62,935 SNPs. Using principal component analysis, Neighbor-Net and Admixture algorithms, the studied breeds were characterized according to their genomic architecture, ancestry and direction of selective breeding. Japanese and Pharaoh breeds had the smallest number and length of homozygous segments indicating a lower selective pressure. Tuxedo and Texas White breeds showed the highest values of these indicators and genomic inbreeding suggesting a greater homozygosity. We revealed evidence for the integration of genomic and performance data, and our findings are applicable for elucidating the history of creation and genomic variability in quail breeds that, in turn, will be useful for future breeding improvement strategies. Full article
(This article belongs to the Special Issue New Tools for Monitoring Genetic Diversity in Animals)
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<p>Clustering reconstruction of the eight breeds studied using the IPI-based pairwise Euclidean distances. (<b>a</b>,<b>b</b>) PCA plots for first (PC1) and second (PC2) components (<b>a</b>), and for first (PC1) and third (PC3) components (<b>b</b>) using the Phantasus web tool [<a href="#B53-animals-13-03439" class="html-bibr">53</a>]. (<b>c</b>) A Neighbor-Joining rootless axial tree built with no proportional edge length and using the Neighbor Joining method [<a href="#B55-animals-13-03439" class="html-bibr">55</a>] and the online T-REX tool [<a href="#B54-animals-13-03439" class="html-bibr">54</a>]. Quail breeds: JAP, Japanese; ENW, English White; ENB, English Black; TUX, Tuxedo; MAG, Manchurian Golden; EST, Estonian; PHA, Pharaoh; TEW, Texas White.</p>
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<p>GBS-based PCA plots for the eight quail breeds studied. (<b>a</b>) View in 3D. (<b>b</b>) Plot composed in the plane of the first (<span class="html-italic">X</span>-axis, PC1) and second (<span class="html-italic">Y</span>-axis, PC2) components. (<b>c</b>) Plot drawn in the plane of the first (<span class="html-italic">X</span>-axis, PC1) and third (<span class="html-italic">Y</span>-axis, PC3) components. Quail breeds: JAP, Japanese; ENW, English White; ENB, English Black; TUX, Tuxedo; MAG, Manchurian Golden; EST, Estonian; PHA, Pharaoh; TEW, Texas White.</p>
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<p>GBS-based PCA plots for the eight quail breeds studied. (<b>a</b>) View in 3D. (<b>b</b>) Plot composed in the plane of the first (<span class="html-italic">X</span>-axis, PC1) and second (<span class="html-italic">Y</span>-axis, PC2) components. (<b>c</b>) Plot drawn in the plane of the first (<span class="html-italic">X</span>-axis, PC1) and third (<span class="html-italic">Y</span>-axis, PC3) components. Quail breeds: JAP, Japanese; ENW, English White; ENB, English Black; TUX, Tuxedo; MAG, Manchurian Golden; EST, Estonian; PHA, Pharaoh; TEW, Texas White.</p>
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<p>Neighbor-Net tree based on pairwise IBS distances. Quail breeds: JAP, Japanese; ENW, English White; ENB, English Black; TUX, Tuxedo; MAG, Manchurian Golden; EST, Estonian; PHA, Pharaoh; TEW, Texas White.</p>
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<p>Admixture-assisted ancestry cluster analysis. (<b>a</b>) CV error calculations for different number of ancestral populations or clusters (from 1 to 9). (<b>b</b>) Horizontal view at K = 3 and 5 clusters. (<b>c</b>) Circular view for K equaling 2 to 9 clusters. Quail breeds: JAP, Japanese; ENW, English White; ENB, English Black; TUX, Tuxedo; MAG, Manchurian Golden; EST, Estonian; PHA, Pharaoh; TEW, Texas White.</p>
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<p>Admixture-assisted ancestry cluster analysis. (<b>a</b>) CV error calculations for different number of ancestral populations or clusters (from 1 to 9). (<b>b</b>) Horizontal view at K = 3 and 5 clusters. (<b>c</b>) Circular view for K equaling 2 to 9 clusters. Quail breeds: JAP, Japanese; ENW, English White; ENB, English Black; TUX, Tuxedo; MAG, Manchurian Golden; EST, Estonian; PHA, Pharaoh; TEW, Texas White.</p>
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<p>Phylogenetic trees based on <span class="html-italic">F</span><sub>ST</sub> genetic distances characterizing the genetic relationships between the studied quail populations. (<b>a</b>) A reconstructed Neighbor-Net network. (<b>b</b>) A Neighbor-Joining rootless hierarchical horizontal tree built with proportional edge length and using the Neighbor Joining method [<a href="#B55-animals-13-03439" class="html-bibr">55</a>] and the online T-REX tool [<a href="#B54-animals-13-03439" class="html-bibr">54</a>]. Quail breeds: JAP, Japanese; ENW, English White; ENB, English Black; TUX, Tuxedo; MAG, Manchurian Golden; EST, Estonian; PHA, Pharaoh; TEW, Texas White.</p>
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<p>Descriptive statistics of the runs of homozygosity (ROH) according to ROH length class. (<b>a</b>) Overall mean length of ROHs (<span class="html-italic">Y</span>-axis) according to ROH length class (<span class="html-italic">X</span>-axis; 0.5–2, 2–4, 4–8 and 8–16 Mb) (<b>b</b>). Mean number of ROHs (<span class="html-italic">Y</span>-axis) according to ROH length class (<span class="html-italic">X</span>-axis; 0.5–2, 2–4, 4–8 and 8–16 Mb). Quail breeds: JAP, Japanese; ENW, English White; ENB, English Black; TUX, Tuxedo; MAG, Manchurian Golden; EST, Estonian; PHA, Pharaoh; TEW, Texas White.</p>
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<p>Assessed degree of divergence and the level of gene flow between the studied breeds using 30 iterations. (<b>a</b>) Rooted maximum likelihood tree with one migration event. Cut length 10 s.e. corresponds to ten times the average standard error (s.e.) estimated from the sample covariance matrix. Estimated gene flow is shown by an arrow pointing from a donor population (PHA) to a recipient one (TEW) and is colored red in proportion to the intensity of the gene flow. (<b>b</b>) Residual matrix derived from the TreeMix analysis for a single migration event expressed as the number of standard error deviations for the observations in the respective breeds. (<b>c</b>) Plot representing the proportion of variance (<span class="html-italic">f</span>-index) in the sample covariance matrix (¶<span class="html-italic">W</span>) accounted for by the model covariance matrix (<span class="html-italic">W</span>) as a function of the number of migration events. Quail breeds: JAP, Japanese; ENW, English White; ENB, English Black; TUX, Tuxedo; MAG, Manchurian Golden; EST, Estonian; PHA, Pharaoh; TEW, Texas White.</p>
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11 pages, 673 KiB  
Article
Dietary Oyster Mushroom (Pleurotus ostreatus) Waste Inhibits Experimentally Induced Eimeria tenella Challenge in Japanese Quails Model
by Jamal Abdul Nasir, Naila Chand, Shabana Naz, Ibrahim A. Alhidary, Rifat U. Khan, Sajida Batool, Noha T. Zelai, Gianluca Pugliese, Vincenzo Tufarelli and Caterina Losacco
Animals 2023, 13(21), 3421; https://doi.org/10.3390/ani13213421 - 4 Nov 2023
Cited by 6 | Viewed by 1559
Abstract
The aim of this study was to investigate the potential of dietary 3% oyster mushroom (Pleurotus ostreatus) waste in enhancing the anticoccidial effects in broilers challenged with Eimeria tenella infection. The experiment involved a total of 600 Japanese quails, raised from [...] Read more.
The aim of this study was to investigate the potential of dietary 3% oyster mushroom (Pleurotus ostreatus) waste in enhancing the anticoccidial effects in broilers challenged with Eimeria tenella infection. The experiment involved a total of 600 Japanese quails, raised from one to thirty-five days of age, which were divided into four treatment groups. These included a negative control group that received a basal diet (BD) without any anticoccidial or antibiotic supplementation in the non-challenged birds (negative control, NC); a positive control (PC) group consisting of NC birds challenged with E. tenella; a group that received the BD with an anticoccidial drug (standard); and a group that received the BD supplemented with 3% waste from oyster mushrooms (3% Pleurotus ostreatus). The results showed that the feed intake, body weight gain, and feed efficiency were significantly lower in the PC (p < 0.05). However, the growth traits were similar in the standard and 3% Pleurotus ostreatus-treated groups. Similarly, there was no difference (p < 0.05) in the mortality rate, oocyst count in the feces, and lesion score between the standard and 3% Pleurotus ostreatus groups. Based on intestinal histology evaluation, the villi height and width were significantly higher in the standard and 3% Pleurotus ostreatus-treated groups compared to those of the PC (p < 0.01). In conclusion, it was found that 3% Pleurotus ostreatus effectively mitigated the low growth rate of Japanese quails induced by coccidial infection. Full article
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<p>(<b>A</b>) Photomicrograph of cecal villi in unchallenged and untreated Japanese quails (NC), displaying nearly normal villi. (<b>B</b>) Photomicrograph of cecal villi in quails challenged with <span class="html-italic">E. tenella</span> and treated with 3% <span class="html-italic">Pleurotus ostreatus</span> stem waste, revealing no significant lesions related to coccidiosis, but with slight infiltration of reactive cells in the lamina propria. A few merozoites/schizonts were observed (*); (<b>C</b>) Photomicrograph of cecal villi in quails challenged with <span class="html-italic">E. tenella</span> and treated with Amprolium (standard), showing villi similar to those in the unchallenged group (NC). (<b>D</b>) Photomicrograph of cecal villi in quails challenged with <span class="html-italic">E. tenella</span> (PC), displaying the erosion and desquamation of crypt epithelia, along with infiltration of reactive cells in the lamina propria. Merozoites/schizonts were also observed in the crypt of the villi. merozoites/schizont in the cecal crypt (*). Bar: 200 µm.</p>
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14 pages, 1366 KiB  
Article
Supplementation of Japanese Quail (Coturnix coturnix japonica) Breeders with Tagetes erecta Flower Extract and Vitamin E Improves the Oxidative Status of Embryos and Chicks
by Lenilson Fonseca Roza, Evandro Menezes de Oliveira, Lidiane Staub, Tainara Ciuffi Euzébio Dornelas, Paula Toshimi Matumoto Pintro, Danielle Aparecida Munhos Hermoso, Emy Luiza Ishii Iwamoto, Alice Eiko Murakami and Tatiana Carlesso Santos
Poultry 2023, 2(4), 449-462; https://doi.org/10.3390/poultry2040034 - 24 Oct 2023
Cited by 1 | Viewed by 1858
Abstract
The effects of Tagetes erecta flower extract (TFE) and increasing levels of vitamin E (VE) in the diet of Japanese quail breeders on progeny performance and oxidative status were studied. Methods: 480 Japanese quail breeders were distributed in a completely randomized design with [...] Read more.
The effects of Tagetes erecta flower extract (TFE) and increasing levels of vitamin E (VE) in the diet of Japanese quail breeders on progeny performance and oxidative status were studied. Methods: 480 Japanese quail breeders were distributed in a completely randomized design with five treatments and twelve replications of six females and two males each. A control diet (25 mg/kg VE) and four diets supplemented with TFE (3 g/kg) and VE (25, 100, 175, or 250 mg/kg) were used. Fresh yolk samples and the yolk sac and liver from embryos (11 and 15 days) and chicks (hatch and 3 days) were analysed. Data were subjected to ANOVA, a regression linear model, and contrast tests and the level of significance was set at p < 0.05. Results: TF and VE in the maternal diet improved the amount of alfa-tocopherol and total carotenoid content in the yolk. TFE + VE reduced lipid peroxidation and improved the oxidative status in the fresh yolk, in the embryo and chick yolk, and in the liver. Liver superoxide dismutase activity in hatched chicks increased linearly with the VE level and was not altered by TFE. Maternal diets did not influence progeny performance (1 to 28 days) or the relative expression of superoxide dismutase or glutathione peroxidase genes in the liver of chicks. Conclusions: TFE is an effective antioxidant in fresh eggs and supplementation of 3 g/kg TFE and high levels of VE in quail breeders improves the oxidative status of embryos and newly hatched chicks. Full article
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<p>α-Tocopherol concentrations (µg/g) (mean ± standard deviation) in fresh egg yolk (<b>a</b>) and liver (<b>b</b>) of 1-day-old chicks from Japanese quail breeders supplemented with different levels of vitamin E associated or not with Tagetes erecta flower extract. ns: not significant (<span class="html-italic">p</span> &gt; 0.05) for contrast test between 25VE vs. 25VE + TFE. (<b>a</b>): ŷ = 22.3989 + 0.78338VE (R<sup>2</sup> = 0.93; <span class="html-italic">p</span> &lt; 0.001, SEM = 13.07); (<b>b</b>): ŷ = −2.40333 + 3.72373VE (R<sup>2</sup> = 0.94; <span class="html-italic">p</span> &lt; 0.001, SEM = 51.17).</p>
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<p>Total carotenoid content (µg/g β-carotene equivalents) in freeze-dried egg yolk (n = 4 pools of 5 eggs per treatment) from Japanese quail breeders supplemented or not with 3 g/kg <span class="html-italic">Tagetes erecta</span> flower extract. * Means different from each other according to the F test (<span class="html-italic">p</span> &lt; 0.001; SEM = 0.844).</p>
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<p>Superoxide dismutase activity in the liver of chicks from Japanese quail breeders supplemented with different levels of VE associated or not with <span class="html-italic">Tagetes erecta</span> flower extract. ŷ = 0.5805 + 0.00141VE (R<sup>2</sup> = 0.68; <span class="html-italic">p</span> &lt; 0.001, SEM = 0.08). ns: not significant (<span class="html-italic">p</span> &gt; 0.05) for contrast test between 25VE vs. 25VE + TFE.</p>
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<p>Real-time quantitative PCR of the liver of 1-day-old chicks from Japanese quail breeders supplemented with different levels of vitamin E associated or not with <span class="html-italic">Tagetes erecta</span> flower extract. Relative gene expression: (<b>A</b>) glutathione peroxidase (<span class="html-italic">GPX7</span>); (<b>B</b>) superoxide dismutase (<span class="html-italic">SOD1</span>).</p>
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17 pages, 3978 KiB  
Article
Molecular Cloning, Expression Analyses, and Physiological Roles of Cathelicidins in the Bursa of Fabricius of the Japanese Quail, Coturnix japonica
by Takumi Ikeda, Hirotada Kondo, Daiki Nunomura, Genki Sato, Machi Ito, Nanako Yamanaka, Shawichi Iwamuro, Itaru Hasunuma, Sakae Kikuyama and Tetsuya Kobayashi
Antibiotics 2023, 12(8), 1341; https://doi.org/10.3390/antibiotics12081341 - 19 Aug 2023
Viewed by 1403
Abstract
Antimicrobial peptides (AMPs) act directly on pathogens and maintain the anti-inflammatory effects and activation of immunocompetent cells. Therefore, the activation of the immune system in poultry via the elevation of endogenous AMPs has been attempted. In this study, we focused on the host [...] Read more.
Antimicrobial peptides (AMPs) act directly on pathogens and maintain the anti-inflammatory effects and activation of immunocompetent cells. Therefore, the activation of the immune system in poultry via the elevation of endogenous AMPs has been attempted. In this study, we focused on the host defense mechanisms in the bursa of Fabricius (BF) of Japanese quail, cloned the cDNA of cathelicidin (CATH)-1 to -3, and analyzed their expression sites. In situ hybridization experiments revealed the mRNA expression of the CATHs in the interfollicular epithelium surrounding the lumen of the quail BF, which suggests that each CATH may exert its antimicrobial action directly in the BF. The intravenous injection of bacterial lipoteichoic acid and lipopolysaccharide endotoxins into the quail promoted the mRNA expression of CATH-1 and CATH-3 in the BF. The addition of CATH-1 or CATH-2 at the time of the antigen injection into mice resulted in antiserum with high antibody titers. Ad libitum administration of butyrate, a short-chain fatty acid, in the drinking water induced an increase in CATH-2 mRNA expression in the BF under certain conditions. These results may improve the defense mechanisms of quail by stimulating CATH expression in the BF through their diet. Full article
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<p>Nucleotide and deduced amino acid sequences of CATH precursor cDNA that was cloned from <span class="html-italic">Coturnix japonica</span> BF (<b>A</b>–<b>C</b>) and a comparison and schematic alignment of amino acid sequences of CATH-1 to -3 deduced from <span class="html-italic">C. japonica</span> (C.j.) BF cDNA, <span class="html-italic">C. japonica</span> genome DNA, <span class="html-italic">C. coturnix</span> (C.c.) spleen cDNA, and <span class="html-italic">G. gallus</span> (G.g.) BF cDNA (<b>D</b>–<b>F</b>). In the nucleotide sequences, the forward primer-derived sequences are underlined, poly-adenylation signals are in boxes, forward and reverse primers for real-time RT-PCR are in shown in red, and the stop codons are marked with asterisks (*). For the amino acid sequences (<b>A</b>–<b>C</b>), the putative signal peptide, cathelin, and mature CATH sequences are shadowed in gray, light blue, and yellow, respectively, and the sequences used for the antibody production are underlined. In (<b>D</b>–<b>F</b>), different amino acid residues are shadowed in red.</p>
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<p>Effects of various concentrations of synthetic cjCATH-1 on the growth of Gram-negative bacteria <span class="html-italic">E. coli</span> (<b>A</b>), Gram-positive bacteria <span class="html-italic">S. aureus</span> (<b>B</b>), and fungi <span class="html-italic">C. albicans</span> (<b>C</b>). Cells from each bacterial or fungal strain were incubated with serially diluted cjCATH-1 for 18 h at 35 °C. Bars and vertical bars represent the means ± standard error of the mean (SEM) (<span class="html-italic">n</span> = 4). In all panels, the values of the lowercase letters sharing the same superscripts are not significantly different (<span class="html-italic">p</span> ≥ 0.05).</p>
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<p>In situ hybridization of CATH-1, -2, and -3 precursors with sense (<b>A</b>–<b>C</b>) and antisense (<b>D</b>–<b>I</b>) mRNA probes in the Japanese quail BF. (<b>A</b>,<b>D</b>,<b>G</b>) CATH-1, (<b>B</b>,<b>E</b>,<b>H</b>) CATH-2, and (<b>C</b>,<b>F</b>,<b>I</b>) CATH-3. Boxes in (<b>D</b>–<b>F</b>) indicate the areas that are shown as enlarged views in (<b>G</b>–<b>I</b>), respectively. The hybridization signals of cjCATH-1, -2, and -3 were detected in the IFE, and cjCATH-2 was found in the follicles. IFE, inter-follicular epithelium; FAE, follicle-associated epithelium. Scale bars, 100 μm in (<b>A</b>–<b>F</b>); 20 μm in (<b>G</b>–<b>I</b>).</p>
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<p>Immunohistochemical analysis of CATH-1 (<b>A</b>,<b>C</b>) and CATH-2 (<b>B</b>,<b>D</b>) in the BF of Japanese quail. Sections were pre-treated with 4% skimmed milk-phosphate buffered saline (PBS) for 1 h at 23 °C and then incubated with anti-cjCATH-1 (<b>A</b>) or anti-cjCATH-2 (<b>C</b>) antiserum overnight at 4 °C. Antigen-preabsorbed anti-cjCATH-1 (<b>C</b>) or anti-cjCATH-2 (<b>D</b>) antiserum were used as negative controls. After rinsing with PBS, the sections were incubated with FITC-conjugated donkey anti-guinea pig IgG (<b>A</b>,<b>C</b>) or AlexaFluor568 conjugated goat anti-rabbit IgG (<b>B</b>,<b>D</b>) for 2 h at 23 °C. Fluorescent signals of CATH-1 and CATH-2 were detected in the IFE, and CATH-2 was also detected in the follicles. IFE, inter-follicular epithelium; FAE, follicle-associated epithelium. Scale bars, 100 μm in (<b>A</b>,<b>C</b>), 20 μm in (<b>B</b>), and 150 μm in (<b>D</b>).</p>
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<p>Time course effects of lipoteichoic acid (LTA) (<b>A</b>) and lipopolysaccharide (LPS) (<b>B</b>) on the steady-state levels of CATH-1, -2, and -3 mRNA in the quail BF. Male quails (<span class="html-italic">n</span> = 3–12) were injected with 1 mg/kg body weight of LTA or LPS through the brachial vein. PBS was used as the negative control. After 2, 4, and 8 h, BF specimens were individually collected, and the total RNA was extracted from each sample. The amounts of CATH-1, -2, and -3 mRNA were analyzed by quantitative real-time PCR, and the values were normalized to β-actin mRNA. Each column represents the average of the experiments performed in duplicate. In all panels, values of lowercase letters with the same superscripts are not significantly different (<span class="html-italic">p</span> ≥ 0.05).</p>
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<p>Effect of DEX on the LTA- and LPS-inducible CATH-1 (<b>A</b>) and CATH-3 (<b>B</b>) mRNA expression levels and the expression of TLR (<b>C</b>) and GR (<b>D</b>) mRNA in the quail BF. Male quails (<span class="html-italic">n</span> = 6–13) were injected with LTA (1 mg/kg body weight; BW) alone, LPS (1 mg/kg BW) alone, DEX (2 mg/kg BW) alone, or a combination of LTA or LPS and DEX. After 4 h, the total RNA was extracted from the BF, and the amounts of CATH-1 and CATH-3 mRNA were analyzed. Specimens of total RNA prepared from control animals were also subjected to RT-PCR using specific primers for TLR4, TLR2t1, and TRL2t2 (<b>C</b>) or GR (<b>D</b>), and the reaction products were separated by agarose gel electrophoresis. In panel (<b>A</b>,<b>B</b>), values of lowercase letters with the same superscripts are not significantly different (<span class="html-italic">p</span> ≥ 0.05).</p>
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<p>Promotive effects on the antibody production in mice through the co-administration of cjCATH-1 and cjCATH-2 with an antigen. Female mice (<span class="html-italic">n</span> = 3) were given an intraperitoneal injection of 500 µg of OVA in 500 µL of Hank’s Balanced Salt Solutions with 10 or 100 µg of cjCATH-1 or cjCATH-2, respectively. After 14 days, another OVA injection was administrated. After more than six days later, blood was collected from the tail vein of the mice, and the titers of the anti-OVA antibodies in the serum were measured. These titers were calculated from the relative levels to those of the control (0 µg OVA on the day 20). Values of lowercase letters with the same superscripts are not significantly different (<span class="html-italic">p</span> ≥ 0.05).</p>
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<p>Effect of ad libitum consumption of short-chain fatty acids (SCFAs) on each of the CATH subtype mRNA expression levels (<b>A</b>), effects of treatment period (<b>B</b>), administrated dosage for 48 h (<b>C</b>), and butyrate receptor antagonist GLPG0974 (<b>D</b>) on the levels of CATH-2 mRNA expression in the quail BF time-course effects, dose-dependent effects, and in the quail BF. Male quails (<span class="html-italic">n</span> = 6–12) were provided with 0.1% butyrate (<b>A</b>,<b>B</b>,<b>D</b>), a type of SCFA, as drinking water ad libitum. After treatment, the BF total RNA specimens were subjected to real-time RT-PCR to measure the levels of the CATHs. The expression levels were calculated from the relative levels to those of the control (0% butyrate). In all panels, values of lowercase letters with the same superscripts are not significantly different (<span class="html-italic">p</span> ≥ 0.05).</p>
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