Nuclei Isolation from Ocular Tissues of the Embryonic Chicken for Single-Nucleus Profiling.

The generation of quality data from a single-nucleus profiling experiment requires nuclei to be isolated from tissues in a gentle and efficient manner. Nuclei isolation must be carefully optimized across tissue types to preserve nuclear architecture, prevent nucleic acid degradation, and remove unwanted contaminants. Here, we present an optimized workflow for generating a single-nucleus suspension from ocular tissues of the embryonic chicken that is compatible with various downstream workflows. The described protocol enables the rapid isolation of a high yield of aggregate-free nuclei from the embryonic chicken eye without compromising nucleic acid integrity, and the nuclei suspension is compatible with single-nucleus RNA and ATAC sequencing. We detail several stopping points, either via cryopreservation or fixation, to enhance workflow adaptability. Further, we provide a guide through multiple QC points and demonstrate proof-of-principle using two commercially available kits. Finally, we demonstrate that existing in silico genotyping methods can be adopted to computationally derive biological replicates from a single pool of chicken nuclei, greatly reducing the cost of biological replication and allowing researchers to consider sex as a variable during analysis. Together, this tutorial represents a cost-effective, simple, and effective approach to single-nucleus profiling of embryonic chicken eye tissues and is likely to be easily modified to be compatible with similar tissue types.

Precooked state based on protein denaturation kinetics impacts moisture status, protein oxidation and texture of prepared chicken breast.

The quality of meat in prepared dishes deteriorates due to excessive protein denaturation resulting from precooking, freezing, and recooking. This study aimed to link the precooked state with chicken breast's recooked quality. Cooked Value (CV), based on protein denaturation kinetics, was established to indicate the doneness of meat during pre-heating. The effects of CVs after pre-heating on recooked qualities were investigated compared to fully pre-heated samples (control). Mild pre-heating reduced water migration and loss. While full pre-heating inhibited protein oxidation during freezing, intense oxidation during pre-heating led to higher oxidation levels. Surface hydrophobicity analysis revealed that mild pre-heating suppressed aggregation during recooking. These factors contributed to a better texture and microstructure of prepared meat with mild pre-heating. Finally, a potential mechanism of how pre-heating affects final qualities was depicted. This study underlines the need for finely controlling the industrial precooking process to regulate the quality of prepared meat.

Extraction and characterization of valuable compounds from chicken sternal cartilage: Type II collagen and chondroitin sulfate.

Type II collagen (Col II) and chondroitin sulfate (CS) are the main macromolecules in the extracellular matrix. This study investigated the characteristics of Col II and CS obtained from chicken sternal cartilage (CSC) via enzymatic hydrolysis for various treatment times. For Col II and CS, the highest efficiency of enzymatic hydrolysis was achieved after 24 and 6 h of treatment, respectively. The average molecular weights were α1 chain-130 kDa, ß chain-270 kDa for Col II, and 80.27 kDa for CS. Fourier transform infrared spectroscopy revealed that the Col II samples maintained their triple-helical structure and that the predominant type of CS was chondroitin-4-sulfate. Scanning electron microscopy revealed that the Col II and CS samples possessed fibrillar and clustered structures, respectively. This study suggests that collagen and CS obtained from CSC can be used as promising molecules for application in food and pharmaceutical industries.

The revealing of the Cyto-genotoxic properties (Allium and MTT) and the effect of chicken meat quality of characterized zein-eugenol nanofibers.

Electrospun zein-based eugenol nanofibers (ZEnF) with diameters (148.19-631.52 nm) were fabricated. Thermal degradation was found as <15 % until 300 °C while the nanofiber diffraction pattern presented three main peaks among the 5o and 45o positions. ZEnF was not only evaluated as non-toxic to cells but also possessed anticancer characteristics revealing with the MCF-7 cell line at 800 µg/mL (reduction: 18.08 %) and 1600 µg/mL (reduction: 41.64 %). Allium tests revealed that ZEnF did not have any adverse impact on the health status (chromosomes-DNA) of exposed organisms. Following the nanofiber coating for chicken meat parts (thigh and breast), it was observed up to 1.25 log CFU/g limitation in total viable bacteria counts (p < 0.05). The sensory score (difference: 3.64 in 10 points scoring on the 6th day of the cold storage) and odor score of chicken meat samples were found to be as higher than control samples (p < 0.05).

Comprehensive metabolic profiling of dioxin-like compounds exposure in laying hens: Implications for toxicity assessment.

The evaluation of toxicity related to polychlorinated dibenzo-p-dioxins and furans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (DL-PCBs) is crucial for a comprehensive risk assessment in real-world exposure scenarios. This study employed a controlled feeding experiment to investigate the metabolic effects of dioxin-like compounds (DLCs) on laying hens via feed exposure. Diets enriched with two concentrations (1.17 and 5.13 pg toxic equivalents (TEQ)/g dry weight (dw)) were administered over 14 days, followed by 28 days of clean feed. Metabolomics analyses of blood samples revealed significant metabolic variations between PCDD/Fs and DL-PCBs exposed groups and controls, reflecting the induced metabolic disruption. Distinct changes were observed in sphingosine, palmitoleic acid, linoleate, linolenic acid, taurocholic acid, indole acrylic acid, and dibutyl phthalate levels, implying possible connections between PCDD/Fs and DL-PCBs toxic effects and energy-neuronal imbalances, along with lipid accumulation and anomalous amino acid metabolism, impacting taurine metabolism. Moreover, we identified three differential endogenous metabolites-L-tryptophan, indole-3-acetaldehyde, and indole acrylic acid-as potential ligands for the aryl hydrocarbon receptor (AhR), suggesting their role in mediating PCDD/Fs and DL-PCBs toxicity. This comprehensive investigation provides novel insights into the metabolic alterations induced by PCDD/Fs and DL-PCBs in laying hens, thereby enhancing our ability to assess risks associated with their exposure in human populations.

Selenium deficiency exacerbates ROS/ER stress mediated pyroptosis and ferroptosis induced by bisphenol A in chickens thymus.

Bisphenol A (BPA) is an industrial pollutant that can cause immune impairment. Selenium acts as an antioxidant, as selenium deficiency often accompanies oxidative stress, resulting in organ damage. This study is the first to demonstrate that BPA and/or selenium deficiency induce pyroptosis and ferroptosis-mediated thymic injury in chicken and chicken lymphoma cell (MDCC-MSB-1) via oxidative stress-induced endoplasmic reticulum (ER) stress. We established a broiler chicken model of BPA and/or selenium deficiency exposure and collected thymus samples as research subjects after 42 days. The results demonstrated that BPA or selenium deficiency led to a decrease in antioxidant enzyme activities (T-AOC, CAT, and GSH-Px), accumulation of peroxides (H2O2 and MDA), significant upregulation of ER stress-related markers (GRP78, IER 1, PERK, EIF-2α, ATF4, and CHOP), a significant increase in iron ion levels, significant upregulation of pyroptosis-related gene (NLRP3, ASC, Caspase1, GSDMD, IL-18 and IL-1ß), significantly increase ferroptosis-related genes (TFRC, COX2) and downregulate GPX4, HO-1, FTH, NADPH. In vitro experiments conducted in MDCC-MSB-1 cells confirmed the results, demonstrating that the addition of antioxidant (NAC), ER stress inhibitor (TUDCA) and pyroptosis inhibitor (Vx765) alleviated oxidative stress, endoplasmic reticulum stress, pyroptosis, and ferroptosis. Overall, this study concludes that the combined effects of oxidative stress and ER stress mediate pyroptosis and ferroptosis in chicken thymus induced by BPA exposure and selenium deficiency.

Effects of substitution of crude palm oil with palm-pressed fibre oil on the laying hen's performance, egg production and egg quality traits.

This study aimed to evaluate the effects of substituting a crude palm oil (CPO) diet with palm-pressed fibre oil (PPFO) on laying hens' performance, egg production, carcass characteristic and egg quality. A total of 150 Hisex Brown laying hens were randomly assigned to five treatments: a basal diet containing 4% CPO (T1-control) and basal diets in which CPO was substituted by 25% (T2), 50% (T3), 75% (T4) or 100% (T5) PPFO. These diets were fed to laying hens ad libitum for 16 weeks. Compared to the T1 diet, dietary treatments T2, T3 and T4 had no significant effect (P > 0.05) on the feed intake, body weight gain, feed conversion ratio, egg number, egg production, egg weight and egg mass of the laying hens during the entire experimental period. Nonetheless, a significant (P < 0.05) reduction in feed intake, egg number, egg production and egg mass were observed in hens fed T5 compared to those fed the T1 (control) diet. Except for heart and spleen weights, other carcass characteristics were not affected by treatment (P < 0.05). Skin yellowness and breast meat redness increased significantly (linearly, P < 0.05) with increasing proportions of PPFO. Likewise, yolk colour scores increased in proportion to the dietary level of PPFO (linearly, P < 0.05). These findings suggest that PPFO can be used as a novel, cost-effective lipid source at a level of 3% in layer diets as a substitute for CPO to improve eggs' quality parameters without any harmful effects on laying performance.

High dietary Mucuna pruriens utilis seed meal compromises growth performance, carcass traits, haemato-biochemistry, and meat quality of broilers.

Usage of soyabean meal (SBM) in broiler diets is economically and environmentally unsustainable thus necessitating investigation of alternative protein sources. Therefore, this study investigated effects of incremental inclusion levels of Mucuna pruriens utilis seed meal (MSM) for partial substitution of SBM in broiler diets. In a completely randomized design (CRD), 400 day-old Ross 308 chicks were allotted to 5 iso-caloric-nitrogenous MSM-containing (0, 5, 10, 15, and 20%) dietary treatments. Each treatment was replicated 8 times, with each pen having 10 birds, during starter (d1 - 14), grower (d15 - 28), and finisher (d29 - 42) phases. Results showed that dietary MSM decreased feed intake (FI: quadratic: P < 0.001), body weight gain (BWG: linear: P < 0.001), and feed conversion efficiency (FCE: linear: P < 0.001) as it linearly decreased slaughter weight (SW: P < 0.001), hot carcass weight (HCW: P < 0.001), cold carcass weight (CCW: P < 0.001), dressing percentage (P < 0.001), and breast weight (P < 0.05). In contrast, dietary MSM linearly increased the weights of the liver (P < 0.01), proventriculus (P < 0.001), gizzard (P < 0.001), duodenum (P = 0.01), jejunum (P < 0.001), ileum (P < 0.001), caecum (P < 0.01), and colon (P < 0.01). Also, dietary MSM quadratically increased blood heterophils (P < 0.05) and alkaline phosphatase activity (P < 0.05) of the chickens whilst linearly increasing their serum amylase (P = 0.001) and lipase (P = 0.001) activities and linearly decreasing their serum symmetric dimethylarginine (SDMA: P = 0.001) and cholesterol (P < 0.05). Further, dietary MSM linearly decreased chicken breast meat ultimate pH (P < 0.05) whilst linearly increasing its cooking loss (P < 0.01), drip loss (P < 0.05) and shear force (P < 0.01). In conclusion, dietary MSM compromised growth performance, carcass characteristics, and meat quality of broilers as it increased the weights of their digestive-metabolic organs.

Response of broilers to supplementation of Salvia officinalis L. essential oil in hot ambient conditions.

Heat stress has various detrimental effects on poultry production. The aim of the study was to alleviate the effects of heat stress in broiler production. For this purpose, 288 one-day-old broiler chicks (Ross 308) were obtained from a commercial hatchery and randomly allocated to one of three treatment groups; CON: corn-soybean meal based commercial diet, SEO100: CON with 100 mg/kg Sage essential oil (SEO) and SEO200: CON with 200 mg/kg SEO with 96 birds in each group (4 replicates each) in a completely randomized design under hot ambient temperatures for 42 days. No differences were observed in the body weight, feed intake, and feed conversion ratio at 42 days of age among groups. However, there was a significant increase in 21-day body weight in SEO200 chicks compared to CON. Furthermore, the addition of SEO significantly decreased the mortality rate under heat stress conditions. The total oxidant status value was lower in broiler chickens in which SEO was added to their diets. While the total antioxidant status value was higher in SEO100 chicks, it did not show a linear increase. Additionally, the results demonstrated that the addition of SEO to broiler diets under heat stress did not have a significant effect on inspected meat quality traits, with the exception of the b*(yellowness) value of breast muscle. In conclusion, the results of this study indicate that 200 mg/kg SEO can be added to diets for the welfare of broiler chickens under heat stress conditions for struggling with oxidants and increasing viability. Further research is needed to investigate the antioxidant activity and meat quality of different levels of SEO in hot ambient conditions.

Metabolome and Transcriptome Profiling Reveals Age-Associated Variations in Meat Quality and Molecular Mechanisms of Taihe Black-Bone Silky Fowls.

To explore the changes in meat quality and molecular mechanisms during the growth and development of Taihe black-bone silky fowl, this study employed liquid chromatography-mass spectrometry (LC-MS/MS) metabolomics to elucidate the dynamic changes of key differential metabolites (DMs) affecting meat quality, indicating that chicken at D120 had higher levels of ω-3 polyunsaturated fatty acids (PUFAs), creatine, anserine, and homocarnosine, while D150 had the most stachydrine and D210 had the most acylcarnitines. Additionally, D120 and D180 had more umami and sweet compounds. Furthermore, key metabolic pathways influenced by age included purine metabolism, the pentose phosphate pathway, nicotinate and nicotinamide metabolism, and taurine and hypotaurine metabolism. Transcriptomic identified differential expression genes (DEGs) are predominantly enriched in focal adhesion, the TGF-ß signaling pathway, and the MAPK signaling pathway. Integrated metabolomics and transcriptomics revealed complex regulatory networks of DEGs and DMs in key metabolic pathways. This research enhanced our understanding of the biology of Taihe black-bone silky fowl meat quality, revealing possible biomarkers.

Influence of age at slaughter and sex on carcass characteristics, meat quality, fatty acids, and ribonucleotides in white-tailed yellow native chickens.

This study investigated the effects of age and sex on carcass and meat characteristics of one of Thailand's major indigenous chicken breeds, the white-tailed yellow native chicken (NC). A total of 120 one-day-old NC (60 males and 60 females) were raised, and harvested at either 16, 20 or 24 weeks. The results showed that body, carcass, breast and fillet weights did not differ (P > 0.05) between 16- and 20-week-old NC, but were lower (P < 0.05) than those of 24-week-old NC. Male NC had higher (P < 0.05) body, carcass, wing, back and thigh weights than female NC. Neither sex nor age affected muscle pH, sarcomere length, redness and yellowness, guanosine monophosphate, and hypoxanthine. The interaction between age and sex was significant (P < 0.05) for %dressing, %leg, L*, C14:1, C18:1n9 and C20:4n6. Shear force was lower in 16-week-old NC (P < 0.05). The 24-week-old NC had lower (P < 0.05) C13:0, C16:0, C18:0, C18:2n6t, C20:4n6, C22:6n-3, intramuscular fat and inosine 5'-monophosphate levels and higher (P < 0.05) C18:2n6c, C18:3n-3 and C20:3n-6 levels than the 16- and 20-week-old NC. Male NC had higher (P < 0.05) C13:0-, C14:0-, C18:2n6t-, C20:3n-6- and lower inosine levels than female NC. In conclusion, these data highlight age- and sex-specific differences in carcass and meat quality of NC and provide relevant information to support consumer-oriented decisions on the production, processing and nutritional value of NC.

Prevalence data on chicken diseases in low-resource settings.

The presence of transmissible disease in livestock has a major impact on welfare and economics in animal and public health. A lack of data enables the spread of diseases due to misinformed decision-making on prevention and control. Low-resource settings face challenges in providing data, turning data availability into a development issue. For this study, a large dataset (n = 997) was collected on prevalence and seroprevalence estimates on viral (n = 224), bacterial (n = 83) and parasitic (n = 690) diseases in backyard chickens in low- and middle-income countries (LMICs). These estimates originate from 306 studies identified during the screening phase of a systematic literature review. An attempt was made to classify the studies according to the Food and Agriculture Organization of the United Nations'classification system for family poultry production systems. Of the studies, 98.7% (302/306) focused on a single poultry production system, while 1.3% (4/306) targeted two different production systems. Within the group of studies that covered one production system, 85.4% (258/302) were classified as â€Ëœsmall extensive scavenging or extensive scavenging,'â€Ëœsmall extensive scavenging'and/or â€Ëœextensive scavenging'. In addition, 52% (159/306) of the studies did not report information on chicken breed type. No data were found on any relevant disease for 56.9% (78/137) of LMICs, signifying a potential data gap. Of the estimates on viral and bacterial diseases, 71.0% (218/307) corresponded to diseases notifiable to the World Organisation for Animal Health, highlighting a tendency to measure disease occurrence for diseases relevant to trade. The latter might not necessarily be priority diseases for the producers, however. Furthermore, 72.3% (222/307) of the estimates originate from random samples and could be used to estimate prevalence in backyard chickens using imputation methods, thus bridging the data gap.

Les maladies transmissibles affectant les animaux d'élevage ont un impact majeur sur la santé animale et la santé publique, avec des effets sur le bien-être et sur l'économie. L'absence de données favorise la propagation des maladies puisque les mesures de prévention et de contrôle reposent sur des décisions mal informées. Les contextes faiblement dotés en ressources se heurtent à la difficulté de produire des données, ce qui fait de la disponibilité des données un enjeu de développement. Pour les besoins de la présente étude, un vaste jeu de données (n = 997) a été constitué, regroupant les taux de prévalence et de séroprévalence estimés d'un certain nombre de maladies virales (n = 224), bactériennes (n = 83) et parasitaires (n = 690) affectant les poulets de basse-cour dans les pays à revenu faible et intermédiaire. Ces estimations sont extraites des 306 études retenues lors de la phase de sélection initiale d'un examen systématique de la littérature. Une tentative de classement de ces études a été réalisée en se basant sur la classification des systèmes d'aviculture familiale élaborée par l'Organisation des Nations Unies pour l'alimentation et l'agriculture. Au total, 98,7 % (302/306) des études portaient sur un système unique de production de volailles, les 1,3 % restantes (4/306) portant sur deux systèmes de production différents. Dans le groupe des études couvrant un seul système de production, 85,4 % des élevages étudiés (258/302) relevaient des catégories " petits systèmes extensifs en liberté ou systèmes extensifs en liberté ", " petits systèmes extensifs en liberté " et/ou " systèmes extensifs en liberté ". En outre, dans 52 % des études (159/306), la race des poulets n'était pas précisée. Aucune donnée n'a pu être trouvée concernant les maladies importantes des volailles dans 56,9 % (78/137) des pays à revenu faible ou intermédiaire, ce qui indique un déficit potentiel de données. S'agissant des maladies virales et bactériennes, 71,0 % des estimations (218/307) correspondaient à des maladies à déclaration obligatoire à l'Organisation mondiale de la santé animale, ce qui souligne la tendance à signaler la survenue des maladies ayant une incidence sur les échanges internationaux. Toutefois, ce ne sont pas nécessairement ces maladies qui sont prioritaires pour les éleveurs. D'autre part, 72,3 % (222/307) des estimations provenaient d'échantillons aléatoires et pourraient donc servir à estimer la prévalence chez les poulets de basse-cour en appliquant des méthodes d'imputation, ce qui permettrait de combler les écarts.

La presencia de enfermedades transmisibles en los animales de granja tiene importantes repercusiones en el bienestar y la economía tanto en el ámbito de la sanidad animal como en el de la salud pública. La falta de datos favorece la propagación de enfermedades debido a la toma de decisiones en materia de prevención y control basada en información mal fundada. En los entornos de bajos recursos existen dificultades para el suministro de información, lo que convierte la disponibilidad de datos en un problema de desarrollo. Para este estudio, se recopiló un amplio conjunto de datos (n = 997) sobre estimaciones de prevalencia y seroprevalencia de enfermedades víricas (n = 224), bacterianas (n = 83) y parasitarias (n = 690) en pollos de traspatio en países de ingresos medios y bajos (PIMB). Estas estimaciones provienen de 306 estudios encontrados durante la fase de selección de una revisión bibliográfica sistemática. Se intentó clasificar los estudios según la clasificación de sistemas de producción avícola familiar de la Organización de las Naciones Unidas para la Alimentación y la Agricultura. El 98,7 % de los estudios (302/306) se centraron en un único sistema de producción avícola, mientras que el 1,3 % (4/306) se centraron en dos sistemas de producción diferentes. Dentro del grupo de estudios que abarcaban un solo sistema de producción, el 85,4 % (258/302) se clasificaron como sistema "extensivo de escarbado pequeño o extensivo de escarbado", "extensivo de escarbado pequeño" o "extensivo de escarbado". Además, en el 52 % (159/306) de los estudios no se proporcionó información sobre el tipo de raza de los pollos. No se encontraron datos sobre ninguna enfermedad pertinente para el 56,9 % (78/137) de los PIMB, lo que indica una posible carencia de datos. De las estimaciones sobre enfermedades víricas y bacterianas, el 71,0 % (218/307) correspondían a enfermedades de declaración obligatoria a la Organización Mundial de Sanidad Animal, lo que evidencia una tendencia a medir la aparición de enfermedades pertinentes para el comercio. Sin embargo, estas podrían no ser necesariamente enfermedades prioritarias para los productores. Por otro lado, el 72,3 % (222/307) de las estimaciones proceden de muestras aleatorias y podrían utilizarse para estimar la prevalencia en los pollos de traspatio utilizando métodos de imputación, lo que permitiría subsanar la carencia de datos.

Infectious bronchitis virus vaccination, but not the presence of XCR1, is correlated with large differences in chicken caecal microbiota.

The chicken immune system and microbiota play vital roles in maintaining gut homeostasis and protecting against pathogens. In mammals, XCR1+ conventional dendritic cells (cDCs) are located in the gut-draining lymph nodes and play a major role in gut homeostasis. These cDCs sample antigens in the gut luminal contents and limit the inflammatory response to gut commensal microbes by generating appropriate regulatory and effector T-cell responses. We hypothesized that these cells play similar roles in sustaining gut homeostasis in chickens, and that chickens lacking XCR1 were likely to contain a dysbiotic caecal microbiota. Here we compare the caecal microbiota of chickens that were either heterozygous or homozygous XCR1 knockouts, that had or had not been vaccinated for infectious bronchitis virus (IBV). We used short-read (Illumina) and long-read (PacBio HiFi) metagenomic sequencing to reconstruct 670 high-quality, strain-level metagenome assembled genomes. We found no significant differences between alpha diversity or the abundance of specific microbial taxa between genotypes. However, IBV vaccination was found to correlate with significant differences in the richness and beta diversity of the microbiota, and to the abundance of 40 bacterial genera. In conclusion, we found that a lack of XCR1 was not correlated with significant changes in the chicken microbiota, but IBV vaccination was.

Expression of IL-1ß in transgenic Eimeria necatrix enhances the immunogenicity of parasites and promotes mucosal immunity against coccidiosis.

Anticoccidial vaccines comprising living oocysts of Eimeria tenella, Eimeria necatrix, Eimeria maxima, and Eimeria acervulina are used to control coccidiosis. This study explored the potential of IL-1ß to act as a molecular adjuvant for enhancing the immunogenicity of Eimeria necatrix and mucosal immunity. We engineered E. necatrix to express a functional chIL-1ß (EnIL-1ß) and immunized chickens with oocysts of the wild type (EnWT) and tranegenic (EnIL-1ß) strains, respectively. The chickens were then challenged with EnWT oocysts to examine the immunogenicity-enhancing potential of chIL-1ß. As expected, the oocyst output of EnIL-1ß-immunized chickens was significantly reduced compared to those immunized using EnWT. No difference in body weight gain and lesion scores of EnIL-1ß and EnWT groups was observed. The parasite load in the small intestine and caeca showed that the invasion and replication of EnIL-1ß was not affected. However, the markers of immunogenicity and mucosal barrier, Claudin-1 and avian ß-defensin-1, were elevated in EnIL-1ß-infected chickens. Ectopic expression of chIL-1ß in E. necatrix thus appears to improve its immunogenicity and mucosal immunity, without increasing pathogenicity. Our findings support chIL-1ß as a candidate for development of effective live-oocyst-based anticoccidial vaccines.

Turmeric essential oil improves intestinal integrity, immunological parameters, and performance of broiler chickens under cyclic heat stress.

This study aimed to examine whether dietary supplementation of broiler chickens with turmeric essential could mitigate the effects of cyclic heat stress conditions. Intestinal and immunological parameters and gene expression were evaluated during the grower phase. A total of 320 21-day-old male Cobb 500 broilers were distributed according to a completely randomized design with a 4 (diet) × 2 (environment) factorial arrangement and eight replications of five birds each. Dietary treatments consisted of a basal diet without essential oil (EO, negative control) and three diets containing low (100 mg kg-1), intermediate (200 mg kg-1), or high (300 mg kg-1) levels of turmeric EO. In the heat stress group, dietary supplementation with turmeric EO at 100 and 200 mg kg-1 improved body weight, feed conversion, breast yield, and relative liver weight. These supplementation levels reduced villus width, increased villus/crypt ratio, reduced the H/L ratio, and improved hepatic (HSP70 and SREBP1) and intestinal (OCLN) gene expression in birds under heat stress. These findings support the hypothesis that turmeric EO can be used to improve or restore intestinal integrity, modulate inflammation parameters, and, consequently, enhance the performance of broilers challenged by cyclic heat stress.

Molecular phylogenetic analysis and seasonal dynamics of Eimeria species infecting broilers of Kashmir, India.

Globally, the poultry industry is seriously threatened by coccidiosis caused by various species of Eimeria. This protozoan parasite inhabits the epithelial lining of the gastrointestinal tract of poultry globally and can cause serious clinical disease. The present study was carried out on poultry farms located in various regions of Kashmir, India, to investigate the prevalence and phylogenetic relationships of Eimeria species affecting broiler chickens. Over a period of one year, fecal samples were collected from 60 poultry farms in Kashmir and morphological and molecular techniques were employed for Eimeria species identification. Results revealed a high prevalence of coccidiosis, with 58.3% (35/60) of farms positive for Eimeria. The most prevalent species were E. tenella (31/35, 88.6%) followed by E. acervulina (25/35, 71.4%), E. maxima (19/35, 54.3%), E. mitis (18/35, 51.4%), and E. necatrix (9/35, 25.7%). Seasonal variation in prevalence was also observed, with the highest rates in autumn (86.7%) and summer (66.7%). Additionally, younger birds (3-4 weeks) exhibited higher infection rates (85.7%) compared to older birds (57.9%) (5-6 weeks). Mixed infection was found in 94.2% (33/35) of positive farms. Phylogenetic analysis using ITS1 sequences confirmed species clustering and revealed evolutionary relationships among Eimeria species. E. tenella and E. necatrix formed a distinct clade, while E. acervulina formed another. The study underscores the importance of molecular techniques in accurate species identification and provides valuable insights into the epidemiology of coccidiosis in poultry in Kashmir. Effective control strategies, including vaccination and improved management practices, are necessary to mitigate the economic losses associated with this widespread poultry disease.

Single nucleus/cell RNA-seq of the chicken hypothalamic-pituitary-ovarian axis offers new insights into the molecular regulatory mechanisms of ovarian development.

The hypothalamic-pituitary-ovarian (HPO) axis represents a central neuroendocrine network essential for reproductive function. Despite its critical role, the intrinsic heterogeneity within the HPO axis across vertebrates and the complex intercellular interactions remain poorly defined. This study provides the first comprehensive, unbiased, cell type-specific molecular profiling of all three components of the HPO axis in adult Lohmann layers and Liangshan Yanying chickens. Within the hypothalamus, pituitary, and ovary, seven, 12, and 13 distinct cell types were identified, respectively. Results indicated that the pituitary adenylate cyclase activating polypeptide (PACAP), follicle-stimulating hormone (FSH), and prolactin (PRL) signaling pathways may modulate the synthesis and secretion of gonadotropin-releasing hormone (GnRH), FSH, and luteinizing hormone (LH) within the hypothalamus and pituitary. In the ovary, interactions between granulosa cells and oocytes involved the KIT, CD99, LIFR, FN1, and ANGPTL signaling pathways, which collectively regulate follicular maturation. The SEMA4 signaling pathway emerged as a critical mediator across all three tissues of the HPO axis. Additionally, gene expression analysis revealed that relaxin 3 (RLN3), gastrin-releasing peptide (GRP), and cocaine- and amphetamine regulated transcripts (CART, also known as CARTPT) may function as novel endocrine hormones, influencing the HPO axis through autocrine, paracrine, and endocrine pathways. Comparative analyses between Lohmann layers and Liangshan Yanying chickens demonstrated higher expression levels of GRP, RLN3, CARTPT, LHCGR, FSHR, and GRPR in the ovaries of Lohmann layers, potentially contributing to their superior reproductive performance. In conclusion, this study provides a detailed molecular characterization of the HPO axis, offering novel insights into the regulatory mechanisms underlying reproductive biology.

Evolution and biological characteristics of H11 avian influenza viruses isolated from migratory birds and pigeons.

The emergence of novel avian influenza reassortants in wild birds in recent years is a public health concern. However, the viruses that circulate in migratory birds are not fully understood. In this study, we summarized and categorized global H11 avian influenza viruses and reported that waterfowl and shorebirds are the major reservoirs of the identified H11 viruses. The surveillance data of the 35,749 faecal samples collected from wild bird habitats in eastern China over the past seven years revealed a low prevalence of H11 viruses in birds, with a positive rate of 0.067% (24 isolates). The phylogenetic analysis of the twenty viruses indicated that H11 viruses have undergone complex reassortment with viruses circulating in waterfowl and shorebirds. These tested viruses do not acquire mammalian adaptive mutations in their genomes and preferentially bind to avian-type receptors. Experimental infection studies demonstrated that the two tested H11N9 viruses of wild bird origin replicated and transmitted more efficiently in ducks than in chickens, whereas the pigeon H11N2 virus isolated from a live poultry market was more adapted to replicate in chickens than in ducks. In addition, some H11 isolates replicated efficiently in mice and caused body weight loss but were not lethal. Our study revealed the role of waterfowl and shorebirds in the ecology and evolution of H11 viruses and the potential risk of introducing circulating H11 viruses into ducks or chickens, further emphasizing the importance of avian influenza surveillance at the interface of migratory birds and poultry.

Improved surrogate eggshell incubation system for assisted reproductive technologies such as cloning, genome modification, and de-extinction efforts.

A germinal disc located on the egg yolk surface drives genetic modification. Windowed and surrogate eggshell incubation methods have been developed, but these exhibit limited abilities to generate transgenic chickens. In the present study, we investigated the frequency of observing the germinal disc according to the preincubation positioning direction and time and found that it depended on those conditions, but only a few chicks (2.8-5.6%) hatched using the windowed method. Then, we attempted to improve surrogate method via one- or two-step procedures. All eggs, including surrogates that were 10 g heavier than the donor eggs, were obtained from a poultry flock of the same age. With the one-step surrogate method, where the donor egg was transferred only once through a 3.5 cm hole on the point end, at the first day of preincubation, into the surrogate egg, the survival rate at day 4 was 30.8%, and the hatching rate was 11.8%. With the two-step surrogate method (transfer was on the 1st and 4th day of incubation), the survival rate at day 4 was improved to 90.7%, and a hatching rate of 70.0% was achieved. Therefore, this method can be effective for in ovo artificial incubation.

Arsenic trioxide induces innate immune response and inflammatory response in chicken liver via cGAS-STING/NF-κB pathway.

Arsenic is a toxic metal-like element widely used in the pesticide, preservative and semiconductor industries. However, accumulation of arsenic through the food chain can cause serious damage to animal and human health. However, the toxic mechanism of arsenic-induced hepatotoxicity in chickens is not clear, and the present study aimed to investigate the potential role of cGAS-STING and NF-κB pathways on inflammatory injury in chicken liver. In this study, 75 white-feathered broilers were divided into a control group, a low-dose arsenic group (4 mg/kg) and a high-dose arsenic group (8 mg/kg) to investigate the toxic effects of arsenic on chicken liver. In this study, we found that pathological changes such as inflammatory cell infiltration and vesicular degeneration occurred in the liver when exposed to ATO. Crucially, exposure to ATO triggered the cGAS-STING pathway and markedly raised the levels of mRNA and protein expression of cGAS, STING, TBK1, and IRF7. The type I interferon response was also triggered. Simultaneously, STING induced the activation of the conventional NF-κB signaling pathway and stimulated the expression of genes associated with inflammation, such as IL-6, TNF-α and IL-1ß. In summary, the induction of inflammatory responses via cGAS-STING and NF-κB signaling pathways under high ATO exposure provides new ideas for further studies on the toxicological mechanisms of arsenic.