Avian ovaries develop asymmetrically apart from prey birds, with only the left ovary growing more towards functional organs. Here, we analyze over 135,000 cells from chick\'s left and right ovaries at six distinct embryonic developmental stages utilizing single-cell transcriptome sequencing. We delineate gene expression patterns across 15 cell types within these embryo ovaries, revealing side-specific development. The left ovaries exhibit cortex cells, zygotene germ cells, and transcriptional changes unique to the left side. Differential gene expression analysis further identifies specific markers and pathways active in these cell types, highlighting the asymmetry in ovarian development. A fine-scale analysis of the germ cell meiotic transcriptome reveals seven distinct clusters with gene expression patterns specific to various meiotic stages. The study also identifies signaling pathways and intercellular communications, particularly between pre-granulosa and germ cells. Spatial transcriptome analysis shows the asymmetry, demonstrating cortex cells exclusively in the left ovary, modulating neighboring cell types through putative secreted signaling molecules. Overall, this single-cell analysis provides insights into the molecular mechanisms of the asymmetric development of avian ovaries, particularly the significant role of cortex cells in the left ovary.

OBJECTIVE: To compare the effects of propofol, ketamine-propofol and isoflurane, at similar anesthetic depth, on cardiopulmonary variables in unpremedictated chickens.
METHODS: Prospective, randomized, crossover experimental trial.
METHODS: A total of 10 male Leghorn domestic chickens, aged 3 months and body mass 1.4-2.0 kg.
METHODS: Birds were randomly assigned to each of three anesthetic protocols, 7 days apart: intravenous propofol, intravenous ketamine-propofol or isoflurane. Anesthesia was induced (indicated by loss of righting reflex and tracheal intubation) and maintained with propofol (10 mg kg-1 minute-1, then 1.1 mg kg-1 minute-1), ketamine-propofol (5 mg mL-1 ketamine and 5 mg mL-1 propofol combined; 10 mg kg-1 minute-1, then 1.1 mg kg-1 minute-1) or isoflurane [5% vaporizer setting initially, then end-tidal concentration (Fe\'Iso) of 2%] for 65 minutes. Anesthesia was maintained at a similar anesthetic depth based upon positive or negative responses to toe pinch. Heart rate (HR), respiratory rate (fR), noninvasive arterial blood pressure and arterial blood gases were measured during anesthesia. Propofol or ketamine-propofol infusion rates and Fe\'Iso required to prevent movement in response to a noxious stimulus and recovery times were recorded.
RESULTS: Anesthesia induction dose was 9.0 ± 0.8 (mean ± SD) and 12.2 ± 0.3 mg kg-1 for propofol and ketamine-propofol, respectively. Propofol and ketamine-propofol infusion rates and Fe\'Iso required to prevent movement in response to the noxious stimulus were 0.88 ± 0.14 mg kg-1 minute-1, 0.92 ± 0.14 mg kg-1 minute-1 and 1.45 ± 0.28%, respectively. Cardiopulmonary variables remained clinically acceptable, but ketamine-propofol was associated with a significantly higher HR (p = 0.0001) and lower fR (p = 0.0001). Time to extubation did not differ among treatments.
CONCLUSIONS: Cardiovascular and respiratory variables were maintained within normal ranges in all treatments. Coadministration of ketamine with propofol significantly reduced the induction and maintenance dose of propofol.

Connexin 43 (Cx43) is a gap junction protein that participates in small molecule exchange between adjacent cells. It is a predominant Cx within the mammalian ovary, where is associated with proper follicle development. The expression and regulation of Cx43 in the chicken ovary is largely unknown. The aim of the present study was to examine the expression of the Cx43 gene (GJA1) and protein as well as the immunolocalization of Cx43 in the laying hen ovary in relation to follicle development, and to examine how tamoxifen (TMX; an estrogen receptor modulator) treatment affects these factors. qRT-PCR and western blotting demonstrated differences in Cx43 mRNA transcript and protein abundances in ovarian white follicles, yellowish follicles, small yellow follicles, and the largest yellow preovulatory follicles (F3-F1). In general, Cx43 was more abundant in hierarchical than prehierarchical follicles and in granulosa cells compared with theca cells. Further, the response to TMX treatment depended on the stage of follicle development and the layer of the follicular wall. Ovarian regression following TMX treatment was accompanied by an increase in Cx43 expression in most ovarian tissues, which may impact the formation and function of Cx43 hemichannels. Overall, our results showed, for the first time, the differences in Cx43 mRNA and protein levels between ovarian follicles, suggesting the potential involvement of this gap junction protein in the regulation of ovarian follicle development and function. In addition, the results indicate a possible role for estradiol in regulation of Cx43 transcription and/or translation in the chicken ovary. Understanding the contribution of Cx43 in mechanisms underlying ovarian follicle development may be of considerable importance for poultry egg production.

Adiponectin (AdipoQ), an adipokine secreted by adipocytes, has been reported to exist widely in various cell types and tissues, including the adenohypophysis of chickens. However, the molecular mechanism by which AdipoQ regulates the function of chicken adenohypophysis remains elusive. In this study, we investigated the effects of AdipoQ on proliferation, apoptosis, secretion of related hormones (FSH, LH, TSH, GH, PRL and ACTH) and expression of related genes (FSHβ, LHβ, GnRHR, TSHβ, GH, PRL and ACTH) in primary adenohypophysis cells of chickens by using real-time fluorescent quantitative PCR (RT-qPCR), cell counting kit-8 (CCK-8), flow cytometry, enzyme-linked immunosorbent assay (ELISA) and Western blot (WB) assays. Our results showed that AdipoQ promoted the proliferation of chicken primary adenohypophysis cells, up-regulated the mRNA expression of proliferation-related genes CDK1, PCNA, CCND1 and P21 (P < 0.05), as well as the increased protein expression of CDK1 and PCNA (P < 0.05). Furthermore, AdipoQ inhibited apoptosis of chicken primary adenohypophysis cells, resulting in down-regulation of pro-apoptotic genes Caspase3, Fas, and FasL mRNA expression, and decreased Caspase3 protein expression (P < 0.05). Moreover, there was an up-regulation of anti-apoptotic gene Bcl2 mRNA and protein expression (P < 0.05). Additionally, AdipoQ suppressed the secretion of FSH, LH, TSH, GH, PRL, and ACTH (P < 0.05), as well as the mRNA expression levels of related genes (P < 0.05). Treatment with AdipoRon (a synthetic substitute for AdipoQ) and co-treatment with RNA interference targeting AdipoQ receptors 1/2 (AdipoR1/2) had no effect on the secretion of FSH, LH, TSH, GH, PRL, and ACTH, as well as the mRNA expression levels of the related genes. This suggests that AdipoQ\'s regulation of hormone secretion and related gene expression is mediated by the AdipoR1/2 signaling axis. Importantly, we further demonstrated that the mechanism of AdipoQ on FSH, LH, TSH and GH secretion is realized through AMPK signaling pathway. In conclusion, we have revealed, for the first time the molecular mechanism by which AdipoQ regulates hormone secretion in chicken primary adenohypophysis cells.

Lambda-cyhalothrin (LCT) is a common pyrethroid insecticide widely used for ectoparasite control and hygiene pest prevention in poultry and this study aimed to investigate the mechanisms of LCT-induced cardiac injury in chickens. Low, medium, and high-dose LCT exposure models in chickens were established and hematoxylin and eosin (H&E) staining, dihydroethidium (DHE) staining, TUNEL staining, immunofluorescence, biochemical analysis, and gene expression analysis were used to study the effects of LCT exposure on the chicken heart. The results showed that LCT exposure increased the serum levels of creatine kinase (CK) and lactate dehydrogenase (LDH), led to muscle fiber breakage and inflammatory cell infiltration and caused cardiac tissue damage. The DHE staining and biochemical analysis revealed that LCT exposure resulted in the excessive accumulation of ROS, decreased activities/levels of catalase (CAT), total superoxide dismutase (T-SOD), and glutathione (GSH), and increased levels of the oxidative damage marker malondialdehyde (MDA). The TUNEL staining indicated that LCT exposure increased apoptosis possibly through the elevated expression of pro-apoptotic genes in the mitochondrial pathway, the reduced expression of anti-apoptotic genes, the upregulation of pro-inflammatory factors and the downregulation of anti-inflammatory factors. Here, LCT exposure significantly inhibited the expression of genes in the Nrf2/HO-1 pathway and activated the expression of genes in the CYP450 enzyme system. Compared to the low-dose group, the high-dose LCT exposure group showed lower levels of apoptosis and inflammation, possibly related to the low oxidative stress levels mediated by the decreased expression of the CYP450 enzyme system. In conclusion, LCT exposure induces oxidative stress, apoptosis, and inflammation in chicken hearts, which may be associated with the inhibition of the Nrf2/HO-1 pathway and activation of the CYP450 enzyme system. This study provides a theoretical basis for the safer use of insecticides in poultry production.

In modern animal husbandry, stress can be viewed as an automatic response triggered by exposure to adverse environmental conditions. This response can range from mild discomfort to severe consequences, including mortality. The poultry industry, which significantly contributes to human nutrition, is not exempt from this issue. Although genetic selection has been employed for several decades to enhance production output, it has also resulted in poor stress resilience. Stress is manifested through a series of physiological reactions, such as the identification of the stressful stimulus, activation of the sympathetic nervous system and the adrenal medulla, and subsequent hormonal cascades. While brief periods of stress can be tolerated, prolonged exposure can have more severe consequences. For instance, extreme fluctuations in environmental temperature can lead to the accumulation of reactive oxygen species, impairment of reproductive performance, and reduced immunity. In addition, excessive noise in poultry slaughterhouses has been linked to altered bird behaviour and decreased production efficiency. Mechanical vibrations have also been shown to negatively impact the meat quality of broilers during transport as well as the egg quality and hatchability in hatcheries. Lastly, egg production is heavily influenced by light intensity and regimens, and inadequate light management can result in deficiencies, including visual anomalies, skeletal deformities, and circulatory problems. Although there is a growing body of evidence demonstrating the impact of environmental stressors on poultry physiology, there is a disproportionate representation of stressors in research. Recent studies have been focused on chronic heat stress, reflecting the current interest of the scientific community in climate change. Therefore, this review aims to highlight the major abiotic stressors in poultry production and elucidate their underlying mechanisms, addressing the need for a more comprehensive understanding of stress in diverse environmental contexts.

The wide distribution and diverse varieties of chickens make them important models for studying genetic adaptation. The aim of this study was to identify genes that alter heat adaptation in commercial chicken breeds by comparing genetic differences between tropical and cold-resistant chickens. We analyzed whole-genome resequencing data of 186 chickens across various regions in Asia, including the following breeds: Bian chickens (B), Dagu chickens (DG), Beijing-You chickens (BY), and Gallus gallus jabouillei from China; Gallus gallus murghi from India; Vietnam native chickens (VN); Thailand native chickens (TN) and Gallus gallus spadiceus from Thailand; and Indonesia native chickens (IN), Gallus gallus gallus, and Gallus gallus bankiva from Indonesia. In total, 5,454,765 SNPs were identified for further analyses. Population genetic structure analysis revealed that each local chicken breed had undergone independent evolution. Additionally, when K = 5, B, BY, and DG chickens shared a common ancestor and exhibited high levels of inbreeding, suggesting that northern cold-resistant chickens are likely the result of artificial selection. In contrast, the runs of homozygosity (ROH) and the ROH-based genomic inbreeding coefficient (FROH) results for IN, TN, and VN chickens showed low levels of inbreeding. Low population differentiation index values indicated low differentiation levels, suggesting low genetic diversity in tropical chickens, implying increased vulnerability to environmental changes, decreased adaptability, and disease resistance. Whole-genome selection sweep analysis revealed 69 candidate genes, including LGR4, G6PC, and NBR1, between tropical and cold-resistant chickens. The genes were further subjected to GO and KEGG enrichment analyses, revealing that most of the genes were primarily enriched in biological synthesis processes, metabolic processes, central nervous system development, ion transmembrane transport, and the Wnt signaling pathway. Our study identified heat adaptation genes and their functions in chickens that primarily affect chickens in high-temperature environments through metabolic pathways. These heat-resistance genes provide a theoretical basis for improving the heat-adaptation capacity of commercial chicken breeds.

A 2-yr-old female Brahma chicken was presented to the Poultry Mobile Clinic of the College of Veterinary Medicine at North Carolina State University with a 3-wk onset of a wet sneeze that progressed to wheezing with a whistle-type sound. Upon observation, a cyst was found above the left clavicle in the area around the crop. The bird was euthanatized due to the progressive and chronic nature of the symptoms. Postmortem examination revealed an ovoid, soft to fluctuant, smooth, pale brown mass (2 × 0.9 × 0.8 cm), encased within the cranial membrane of the left cervical air sac. Histologically, focally expanding the left cervical air sac was a pedunculated, nonencapsulated, well-demarcated, moderately cellular neoplasm that consisted of cuboidal cells predominantly arranged in variably sized cystic structures lined by a single layer of cells. Neoplastic cells have strong cytoplasmic immunolabeling against cytokeratin AE1/AE3. Gross and histologic findings were consistent with an air sac cystadenoma. Primary respiratory neoplasia in birds is infrequent. Air sac carcinomas, adenocarcinomas, and cystadenocarcinomas have been described in Psittaciformes, Columbiformes, Falconiformes, and Cuculiformes. Benign air sac tumors are poorly documented, and detailed descriptions of this neoplasm in poultry literature are lacking.
Reporte de caso- Cistadenoma en los sacos aéreos de un pollo mascota. Una gallina Brahma de dos años fue remitida al Servicio Ambulatorio de Avicultura de traspatio de la Universidad Estatal de Carolina del Norte debido a la presentación de un cuadro clínico de estornudos que progresó a la emisión de ruidos respiratorios tipo silbido en el curso de tres semanas. Se observó un quiste de 2-3 cm de diámetro en el área de la clavícula izquierda alrededor del buche. A dicha ave se le practicó la eutanasia debido a la naturaleza progresiva de los signos. El examen post mortem reveló una masa, ovalada, suave y fluctuante, de color café pálido, de 2 cm × 0.9 cm × 0.8 cm, contenida en la membrana craneal del saco aéreo cervical izquierdo. Histológicamente la pared del saco aéreo cervical izquierdo estaba reemplazada por una neoplasia no encapsulada, pedunculada, bien demarcada, compuesta de células cuboidales organizadas en múltiples estructuras quísticas de tamaño variable y recubiertas por una monocapa celular. Las células neoplásicas poseían una fuerte immunorreactividad citoplasmática para citoqueratina AE1/AE3. Los hallazgos macroscópicos y microscópicos son consistentes con un cistoadenoma de sacos aéreos. En aves, las neoplasias primarias de origen respiratorio son infrecuentes. Carcinomas, adenocarcinomas, y cistoadenocarcinomas de sacos aéreos se han reportado en Psitaciformes, Columbiformes, Falconiformes y Cuculiformes. Los tumores benignos de sacos aéreos han sido escasamente documentados y se carece de descripciones detalladas de estas neoplasias en gallináceas en la literatura.

We previously demonstrated that a prime-boost regime with an infectious bronchitis virus (IBV) Massachusetts (Mass)-type vaccine and recombinant LaSota virus (rLS) coexpressing IBV Arkansas (Ark)-type trimeric spike ectodomain (Se) and granulocyte macrophage colony stimulating factor (GMCSF) enhances heterologous protection against virulent Ark challenge. This study evaluates protection against Ark-type challenge conferred by administering the rLS/ArkSe.GMCSF and the attenuated Mass viruses mixed in the same vial as a combined vaccine. Chickens were vaccinated at day of hatch and challenged at 21 days of age with virulent Ark. Protection conferred by vaccination was assessed by respiratory signs, tracheal virus isolation as well as IBV RNA quantitation, and tracheal histomorphometry. Protection conferred by the combined vaccine was compared to protection induced by a commercial attenuated ArkDPI (Delmarva Poultry Industry) vaccine as well as by the attenuated Mass vaccine alone. Vaccination with the combined vaccine (rLS/ArkSe.GMCSF + Mass) as well as Mass alone provided significantly less protection against Ark challenge compared to the control using attenuated live ArkDPI vaccine. Only ArkDPI-vaccinated chickens exhibited \"sterilizing immunity,\" i.e., no virus isolated from ≥10% of chickens after challenge. Chickens vaccinated with the combined vaccine rLS/ArkSe.GMCSF + Mass showed significantly less tracheal damage than birds vaccinated with the attenuated Mass vaccine alone. In addition, the combined vaccine also resulted in less virus isolation from the trachea. We concluded that the combined vaccine containing the recombinant virus and the attenuated Mass enhanced the cross-protective ability of the attenuated Mass vaccine against heterologous challenge.
Nota de investigación- Protección cruzada conferida por una vacuna combinada que contiene el virus atenuado de la bronquitis infecciosa serotipo Massachusetts y un virus de Newcastle LaSota recombinante que expresa la proteína de la espícula del serotipo Arkansas. Previamente se demostró que un esquema de refuerzo con una vacuna tipo Massachusetts (Mass) del virus de la bronquitis infecciosa (IBV) y con un virus de Newcastle LaSota recombinante (rLS) que co-expresa el ectodominio de pico trimérico del serotipo Arkansas (Ark) del virus de la bronquitis infecciosa (Se) y el factor de estimulación de colonias de granulocitos y macrófagos (GMCSF) (virus rLS/ArkSe.GMCSF) mejora la protección heteróloga contra el desafío con serotipo Arkansas virulento. Este estudio evaluó la protección contra el desafío con el serotipo Arkansas conferida por la administración del virus recombinante rLS/ArkSe.GMCSF y el virus Massachussets atenuado mezclados en el mismo vial como una vacuna combinada. Los pollos se vacunaron el día de la eclosión y se desafiaron a los 21 días de edad con el serotipo Arkansas virulento. La protección conferida por la vacunación se evaluó mediante signos respiratorios, el aislamiento del virus traqueal, cuantificación del ARN del virus de bronquitis y por histomorfometría traqueal. La protección conferida por la vacuna combinada se comparó con la protección inducida por una vacuna comercial atenuada ArkDPI (Delmarva Poultry Industry), y con la vacuna atenuada Massachussets por si sola. La vacunación con la vacuna combinada (rLS/ArkSe.GMCSF + Mass), así como con Massachussets sola, proporcionó una protección significativamente menor contra la exposición con Arkansas en comparación con el control que utilizó la vacuna ArkDPI viva atenuada. Solo los pollos vacunados con ArkDPI exhibieron “inmunidad esterilizante”, es decir, que no se aisló ningún virus en ≥10 % de los pollos después del desafío. Los pollos vacunados con la vacuna combinada rLS/ArkSe.GMCSF + Mass mostraron significativamente menos daño traqueal que las aves vacunadas con la vacuna Massachusetts atenuada sola. Además, la vacuna combinada también resultó en un menor aislamiento del virus de la tráquea. Se concluye que la vacuna combinada que contiene el virus recombinante y el serotipo Massachussets atenuado mejoró la capacidad de protección cruzada de la vacuna de Massachusetts atenuada contra el desafío heterólogo.

Skeletal muscle, which is predominantly constituted by multinucleated muscle fibers, plays a pivotal role in sustaining bodily movements and energy metabolism. Myoblasts, which serve as precursor cells for differentiation and fusion into muscle fibers, are of critical importance in the exploration of the functional genes associated with embryonic muscle development. However, the in vitro proliferation of primary myoblasts is inherently constrained. In this study, we achieved a significant breakthrough by successfully establishing a chicken myoblast cell line through the introduction of the exogenous chicken telomerase reverse transcriptase (chTERT) gene, followed by rigorous G418-mediated pressure screening. This newly developed cell line, which was designated as chTERT-myoblasts, closely resembled primary myoblasts in terms of morphology and exhibited remarkable stability in culture for at least 20 generations of population doublings without undergoing malignant transformation. In addition, we conducted an exhaustive analysis that encompassed cellular proliferation, differentiation, and transfection characteristics. Our findings revealed that the chTERT-myoblasts had the ability to proliferate, differentiate, and transfect after multiple rounds of population doublings. This achievement not only furnished a valuable source of homogeneous avian cell material for investigating embryonic muscle development, but also provided valuable insights and methodologies for establishing primary cell lines.