Search Results (57)

Summer high temperatures (27–38 °C, more than 7 days) readily induce heat stress in late-laying broiler breeders, which impedes offspring growth and development. This study aimed to provide a scientific basis for improving the offspring development of late-laying broiler breeders during summer. Six hundred fertilized eggs from 50-week-old LiFeng broiler breeders were divided into five treatment groups (non-injected, NaCl-injected, 6 mg/egg L-arginine-injected, 3.5 mg/egg L-methionine-injected, 8.4 mg/egg L-leucine-injected), with six replicates per group and 20 eggs per replicate. Embryos were incubated for 21 days and chicks raised for 21 days post-hatch. Methionine injection significantly enhanced hatchability (+5.8%), increased daily chick weight gain (+8.8%), reduced serum urea nitrogen (−53.13%), decreased inflammatory cytokine levels, elevated antioxidant enzyme activities, lowered malondialdehyde content (−47.99%), and suppressed expression of inflammatory and apoptotic pathway genes. The comprehensive effect of methionine was the best among the three amino acids when injected into chicken embryos. Methionine promoted protein synthesis, enhanced antioxidant and anti-inflammatory capacities, and consequently improved chick growth performance. Full article

Rooster behavior and activity are critical for egg fertility and hatchability in broiler and layer breeder houses. Desirable roosters are expected to have good leg health, reach sexual maturity, be productive, and show less aggression toward females during mating. However, not all roosters are desirable, and low-productive roosters should be removed and replaced. The objectives of this study were to apply an object detection model based on deep learning to identify hens and roosters based on phenotypic characteristics, such as comb size and body size, in a cage-free (CF) environment, and to compare the performance metrics among the applied models. Six roosters were mixed with 200 Lohmann LSL Lite hens during the pre-peak phase in a CF research facility and were marked with different identifications. Deep learning methods, such as You Only Look Once (YOLO) models, were innovated and trained (based on a comb size of up to 2500 images) for the identification of male and female chickens based on comb size and body features. The performance matrices of the YOLOv5u and YOLOv11 models, including precision, recall, mean average precision (mAP), and F1 score, were statistically compared for hen and rooster detection using a one-way ANOVA test at a significance level of p < 0.05. For rooster detection based on comb size, YOLOv5lu, and YOLOv11x variants performed the best among the five variants of each model, with YOLOv5lu achieving a precision of 87.7%, recall of 56.3%, and mAP@0.50 of 60.1%, while YOLOv11x achieved a precision of 86.7%, recall of 65.3%, and mAP@0.50 of 61%. For rooster detection based on body size, YOLOv5xu, and YOLOv11m outperformed other variants, with YOLOv5xu achieving a precision of 88.9%, recall of 77.7%, and mAP@0.50 of 82.3%, while YOLOv11m achieved a precision of 89.0%, recall of 78.8%, and mAP@0.50 of 82.6%. This study provides a reference for automatic rooster monitoring based on comb and body size and offers further opportunities for tracking the activities of roosters in a poultry breeder farm for performance evaluation and genetic selection in the future. Full article

Thermal manipulation (TM) during embryogenesis is a promising non-pharmacological strategy to enhance physiological resilience in broiler chickens. This study evaluated the impact of thermal conditioning of fertile eggs on growth performance, inflammatory responses, and molecular stress markers following a post-hatch lipopolysaccharide (LPS) challenge. Fertilized eggs (average weight 62 ± 3 g) were obtained from 35-week-old Indian River broiler breeder hens. A total of 720 eggs were randomly assigned to either the control group (n = 360) or the TM group (n = 360), with each group consisting of two replicates of 180 eggs. Control eggs were maintained under standard incubation conditions (37.8 °C, 56% RH), while TM eggs were subjected to elevated temperature (38.8 °C, 65% RH) for 18 h daily from embryonic day 10 to 18. On post-hatch day 15, control and TM groups were administered either saline or LPS via intraperitoneal (IP) injection. Body weight and temperature, internal organ weights, and splenic mRNA expression levels of inflammatory cytokines, toll-like receptors, transcription factors, and heat shock proteins were assessed. TM did not alter hatchability (p = 0.633), but significantly shortened hatch time (p < 0.05) and improved feed efficiency (p < 0.05). While LPS induced marked inflammatory responses in all birds, those subjected to TM exhibited attenuated proinflammatory cytokine expression, enhanced anti-inflammatory signaling, and differential regulation of stress-associated genes, including nuclear factor kappa B (NF-κB), heat shock protein 70 (HSP70), and heat shock factors (HSFs). These findings suggest that TM during incubation promotes a more regulated immune response and improved stress adaptation post-hatch. This approach offers a potential antibiotic-free intervention to enhance broiler health, performance, and resilience under immunological stress. Full article

Rising global temperatures challenge poultry production by disrupting the cecal microbiota, which is essential for chicken health. Thermal manipulation (TM) during embryogenesis is a potential strategy to enhance thermotolerance in broilers. This study examined TM’s effects on the cecal microbiome, body weight (BW), and body temperature (BT) under chronic heat stress (CHS). Fertile Indian River eggs (n = 800) were incubated under control (37.8 °C, 56% RH) or TM conditions (39 °C, 65% RH for 18 h per day from embryonic day 10 to 18). On post-hatch day 18, male chicks were assigned to either CHS (35 ± 0.5 °C for five days) or thermoneutral conditions (24 ± 0.5 °C). The CHS-TM group showed a significantly higher BW than the CHS-CON group (p < 0.05). Under thermoneutral conditions, TM chicks had a lower BT on day 1 (p < 0.05), while the CHS-TM group exhibited a non-significant BT reduction compared to the CHS-CON group under heat stress (p > 0.05). An analysis of the gut microbiome showed that the beta diversity analysis (PERMANOVA, p < 0.05) indicated distinct microbial shifts. Firmicutes and Bacteroidota dominated the phylum level, with CHS increased Bacilli and Lactobacillus while reducing Lachnospirales in the CHS-TM group. These findings suggest that TM modulates gut microbiota and mitigates BW loss, offering a potential strategy to enhance broilers’ resilience to heat stress. Full article

Fibrosarcomas represent a rare but highly aggressive tumor entity among soft tissue tumors. Due to its rarity, questions regarding its development and pathophysiology remain unclear. The chorioallantoic membrane (CAM) assay represents an easily available method to investigate tumors on a growth membrane, live and in ovo. The following study was established to test whether the growth of fibrosarcoma spheroids on the CAM was possible and to critically review their applicability for downstream investigations. The shells of fertilized chicken eggs were opened and the previously prepared HT1080 cell spheroids (50,000, 75,000, and 100,000 cells per spheroid) were applied to the CAM. After 7 days, tumors were examined for size, weight, and vascularization. After 7 days, 80 of 163 chicken eggs showed sufficient tumor growth. Of these 80 eggs with confirmed tumor growth, 32 (40%) were from the 50,000 spheroid group, 18 (22.5%) were from the 75,000 spheroid group, and 30 (37.5%) were from the 100,000 spheroid group. The 100,000-cell spheroid group exhibited the highest weights, with a mean of 110.7 mg, as well as tumor size expansion. This cell number also showed the highest vascularization rates. Tumor growth of fibrosarcoma spheroids could successfully be initiated on the CAM. Consequently, the CAM assay presents a good base for future studies involving human fibrosarcoma cell spheroids. Full article

This study explores the application of advanced Recurrent Neural Network (RNN) architectures—specifically Long Short-Term Memory (LSTM) and Gated Recurrent Unit (GRU)—for classifying chicken egg fertility based on embryonic development detected in egg images. Traditional methods, such as candling, are labor-intensive and often inaccurate, making them unsuitable for large-scale poultry operations. By leveraging the capabilities of LSTM and GRU models, this research aims to automate and enhance the accuracy of egg fertility classification, thereby contributing to agricultural automation. A dataset comprising 240 high-resolution egg images was employed, resized to 256 × 256 pixels for optimal processing efficiency. LSTM and GRU models were trained to discern fertile from infertile eggs by analyzing the sequential data represented by the pixel rows in these images. The LSTM model demonstrated superior performance, achieving a validation accuracy of 89.58%, significantly surpassing the GRU model (66.67%). Compared to classical methods such as Decision Tree (85%), Logistic Regression (88.3%), the LSTM model demonstrated superior performance, achieving a validation accuracy of 89.58%, significantly surpassing the GRU model (66.67%). Compared to Decision Tree (85%), Logistic Regression (88.3%), SVM (84.57%), K-means (82.9%), and R-CNN (70%), the LSTM model achieved the highest classification accuracy. Unlike classical machine learning approaches that rely on handcrafted features and predefined decision rules, LSTM effectively learns complex sequential dependencies within images, improving fertility classification accuracy in real-world poultry farming applications. In contrast, GRU models, while more computationally efficient, may struggle with generalization under constrained data conditions. This study underscores the potential of advanced RNNs in enhancing the efficiency and accuracy of automated farming systems, paving the way for future research to further optimize these models for real-world agricultural applications. Full article

Understanding the genetic, physiological, and nutritional characteristics of native chickens in South Africa has been significantly hindered by studies over the last ten years. These chickens hold significant economic, social, and cultural importance for South African communities, particularly those marginalized. Despite their reputation for lower egg productivity, they are highly valued for their flavorful meat by consumers. Many local chicken ecotypes and breeds remain undocumented and in danger of going extinct, even though some have been classified. To tackle this issue, the Food and Agriculture Organization has launched an indigenous poultry conservation program. One crucial method employed is assisted reproductive biotechnologies such as cryopreservation, which serves as an ex situ conservation strategy for preserving the germplasm of endangered animals. In avian species, cryopreservation is particularly beneficial for the long-term storage of sperm cells, although it necessitates the use of cryoprotectants to shield sperm cells from cold shock during freezing. However, the use of cryoprotectants can lead to thermal shocks that may damage the sperm cell plasma membrane, potentially reducing viability and fertility. Furthermore, the membranes of avian sperm cells are highly polyunsaturated fatty acids, which can undergo lipid peroxidation (LPO) when reactive oxygen species (ROS) are present. This review focuses on current knowledge and the latest effective strategies for utilizing cryopreservation to conserve semen from indigenous poultry breeds. Full article

A specific-pathogen-free (SPF) chicken colony was maintained with successive groups a month apart in age. The absence of specific pathogens, including chicken anemia virus (CAV), was confirmed through periodic serological tests for each group. However, some groups became CAV seropositive. The procedures of removing seropositive and the adjacent seronegative chickens followed with chemically disinfecting the housing did not halt CAV outbreaks. The full genome sequence of the CAV strain that appeared was closely related to low-virulence isolates in China. The outbreaks of CAV decreased with an increase in the seropositive chicken population, indicating that the progeny is protected from CAV infection by maternal anti-CAV antibodies. The persistence of CAV in erythroid and lymphoid tissues or reproductive tissues from CAV seropositive chickens was examined in chickens of various ages using polymerase chain reaction (PCR). Since a low persistence of CAV was observed in the colony, we isolated eggs from CAV seropositive hens through artificial insemination using semen collected from roosters and confirmed as CAV-free by PCR. Fertilized eggs were transferred to a new SPF facility and used for generating CAV-free progeny. To date, chickens reared in the new facility have been CAV-free for longer than two years. Redirection of eggs from seropositive hens was an effective means of eliminating CAV from chickens. Full article

Background and Objectives: Malignant melanoma (MM) remains one of the most aggressive cancers worldwide, presenting a limited number of therapeutic options at present. Aspirin (ASA), a broadly used non-steroid anti-inflammatory medicine, has recently emerged as a candidate for repurposing in cancer management, due to its therapeutic potential in the treatment of several neoplasms which include MM. Fisetin (FIS) is a flavonoid phytoestrogen instilled with multispectral pharmacological activities, including a potent anti-melanoma property. The present study aimed to assess the potential improved anti-neoplastic effect resulting from the association of ASA and FIS for MM therapy. Materials and Methods: The study was conducted using the A375 cell line as an experimental model for MM. Cell viability was assessed via the MTT test. Cell morphology and confluence were evaluated using bright-field microscopy. The aspect of cell nuclei and tubulin fibers was observed through immunofluorescence staining. The irritant potential and the anti-angiogenic effect were determined on the chorioallantoic membrane of chicken fertilized eggs. Results: The main findings related herein demonstrated that the ASA 2.5 mM + FIS (5, 10, 15, and 20 µM) combination exerted a higher cytotoxicity in A375 MM cells compared to the individual compounds, which was outlined by the concentration-dependent and massive reduction in cell viability, loss of cell confluence, cell shrinkage and rounding, apoptotic-like nuclear features, constriction and disruption of tubulin filaments, increased apoptotic index, and suppressed migratory ability. ASA 2.5 mM + FIS 20 µM treatment lacked irritant potential on the chorioallantoic membrane and inhibited blood-vessel formation in ovo. Conclusion: These results stand as one of the first contributions presenting the anti-melanoma effect of the ASA + FIS combinatorial treatment. Full article

Omics approaches have significantly contributed to our understanding of several aspects of chicken reproduction. This review paper gives an overview of the use of omics technologies such as genomics, transcriptomics, proteomics, and metabolomics to elucidate the mechanisms of chicken reproduction. Genomics has transformed the study of chicken reproduction by allowing the examination of the full genetic makeup of chickens, resulting in the discovery of genes associated with reproductive features and disorders. Transcriptomics has provided insights into the gene expression patterns and regulatory mechanisms involved in reproductive processes, allowing for a better knowledge of developmental stages and hormone regulation. Furthermore, proteomics has made it easier to identify and quantify the proteins involved in reproductive physiology to better understand the molecular mechanisms driving fertility, embryonic development, and egg quality. Metabolomics has emerged as a useful technique for understanding the metabolic pathways and biomarkers linked to reproductive performance, providing vital insights for enhancing breeding tactics and reproductive health. The integration of omics data has resulted in the identification of critical molecular pathways and biomarkers linked with chicken reproductive features, providing the opportunity for targeted genetic selection and improved reproductive management approaches. Furthermore, omics technologies have helped to create biomarkers for fertility and embryonic viability, providing the poultry sector with tools for effective breeding and reproductive health management. Finally, omics technologies have greatly improved our understanding of chicken reproduction by revealing the molecular complexities that underpin reproductive processes. Full article

The sperm-specific phospholipase C zeta (PLCζ) protein is widely considered as the predominant physiological stimulus for initiating the Ca²⁺ release responsible for oocyte activation during mammalian fertilization. The increasing number of genetic and clinical reports that directly link PLCζ defects and/or deficiencies with oocyte activation failure (OAF) necessitates the use of a powerful therapeutic intervention to overcome such cases of male factor infertility. Currently, in vitro fertilization (IVF) clinics treat OAF cases after intracytoplasmic sperm injection (ICSI) with Ca²⁺ ionophores. Despite their successful use, such chemical agents are unable to trigger the physiological pattern of Ca²⁺ oscillations. Moreover, the safety of these ionophores is not yet fully established. We have previously demonstrated that recombinant PLCζ protein can be successfully used to rescue failed oocyte activation, resulting in efficient blastocyst formation. Herein, we produced a maltose binding protein (MBP)-tagged recombinant human PLCζ protein capable of inducing Ca²⁺ oscillations in mouse oocytes similar to those observed at fertilization. Circular dichroism (CD) experiments revealed a stable, well-folded protein with a high helical content. Moreover, the recombinant protein could retain its enzymatic properties for at least up to 90 days after storage at −80 °C. Finally, a chick embryo model was employed and revealed that exposure of fertilized chicken eggs to MBP-PLCζ did not alter the embryonic viability when compared to the control, giving a first indication of its safety. Our data support the potential use of the MBP-PLCζ recombinant protein as an effective therapeutic tool but further studies are required prior to its use in a clinical setting. Full article

Partial least-squares (PLS) regression is a well known chemometric method used for predictive modelling, especially in the presence of many variables. Although PLS was not initially developed as a technique for classification tasks, scientists have reportedly used this approach successfully for discrimination purposes. Whereas some non-supervised learning approaches, including, but not limited to, PCA and k-means clustering, do well in identifying/understanding grouping and clustering patterns in multidimensional data, they are limited when the end target is discrimination, making PLS a preferable alternative. Hyperspectral imaging data on a total of 672 fertilized chicken eggs, consisting of 336 white eggs and 336 brown eggs, were used in this study. Hyperspectral images in the NIR region of the 900–1700 nm wavelength range were captured prior to incubation on day 0 and on days 1–4 after incubation. Eggs were candled on incubation day 5 and broken out on day 10 to confirm fertility. While a total number of 312 and 314 eggs were found to be fertile in the brown and white egg batches, respectively, the total number of non-fertile eggs in the same set of batches was 23 and 21, respectively. Spectral information was extracted from a segmented region of interest (ROI) of each hyperspectral image and spectral transmission characteristics were obtained by averaging the spectral information. A moving-thresholding technique was implemented for discrimination based on PLS regression results on the calibration set. With true positive rates (TPRs) of up to 100% obtained at selected threshold values of between 0.50 and 0.85 and on different days of incubation, the results indicate that the proposed PLS technique can accurately discriminate between fertile and non-fertile eggs. The adaptive PLS approach was, thereby, presented as suitable for handling hyperspectral imaging-based chicken egg fertility data. Full article

The aim of the study was to determine differences in the proteome and peptidome and zinc concentrations in the serum and tissues of chickens supplemented with a multi-strain probiotic and/or zinc glycine chelate in ovo. A total of 1400 fertilized broiler eggs (Ross × Ross 708) were divided into four groups: a control and experimental groups injected with a multi-strain probiotic, with zinc glycine chelate, and with the multi-strain probiotic and zinc glycine chelate. The proteome and peptidome were analyzed using SDS-PAGE and MALDI—TOF MS, and the zinc concentration was determined by flame atomic absorption spectrometry. We showed that in ovo supplementation with zinc glycine chelate increased the Zn concentration in the serum and yolk sac at 12 h post-hatch. The results of SDS-PAGE and western blot confirmed the presence of Cu/Zn SOD in the liver and in the small and large intestines at 12 h and at 7 days after hatching in all groups. Analysis of the MALDI—TOF MS spectra of chicken tissues showed in all experimental groups the expression of proteins and peptides that regulate immune response, metabolic processes, growth, development, and reproduction. Full article

The aim of this study was to investigate the effects of adding perilla seed meal (PSM) to the diet on reproductive performance, egg quality, yolk fatty acids, antioxidant capacity and liver lipid metabolism in breeding hens. A total of 192 31-week-old yellow-feathered hens were randomly divided into 4 treatments with 6 replicates of 8 birds for 8 weeks. The chickens were fed a typical corn–soybean meal diet containing 0% (control), 0.3%, 0.6%, and 1% PSM. The results showed that PSM can change the productivity of laying hens. Adding 0.6% PSM to the feed reduced the mortality rate of chickens. Adding 1% PSM improved the fertilization rate and hatching rate of chickens. Regarding egg quality, the albumen height and Haugh unit were improved in the 0.6% PSM group. The content of MUFAs and PUFAs in the egg yolk was increased in all the PSM groups, while SFAs were only increased in the 0.6% PSM group. Among the indicators related to lipid metabolism, serum GLU decreased in all the PSM groups. The 0.6% PSM group had a reduction in serum and liver TG, as well as reductions in serum LDL-C and ALT. The same results were observed for the abdominal fat percentage in the 0.6% PSM group. Liver lipid metabolism-associated gene expression of FAS and LXRα was decreased in all the PSM groups, and the mRNA expression of ACC and SREBP-1c was significantly reduced in the 0.6% PSM group. HE staining showed that the vacuoles in the liver tissue gradually decreased with increasing PSM doses, especially the 1% PSM dose. Lipid droplets with a similar trend were observed using Oil Red O staining. In the results of the antioxidant capacity test, the serum T-AOC was increased in the 0.6% and 1% PSM groups, and the SOD in both the serum and liver was significantly increased in all the PSM groups. The expression of antioxidant-related genes such as Nrf2, NQO-1, HO-1, CAT and GSH-Px was significantly upregulated in the 1% PSM group. In conclusion, the PSM diet improved the lipid metabolism and antioxidant capacity of breeding hens. PSM reduces mortality and improves fertilization and hatchability in the late laying period of chickens, resulting in greater benefits. We recommend adding 0.6% PSM to layer feed, which improves the physical condition of the hens and brings higher economic benefits. Full article

Fertility rate and hatchability rate are low for all types of double-yolk (DY) eggs in comparison to single-yolk eggs (SY), but these parameters also depend on the number of developing embryos in the egg. The hatchability rate of double-yolk eggs containing two developing embryos (DY2F) is vastly lower than in the case of double-yolk eggs containing only one embryo (DY1F). The aim of the study was to determine the differences between egg fertility rate, hatchability rate, time of embryonic mortality, and embryo malposition during incubation in three types of eggs from Hy-Line Brown hens: SY, DY1F and DY2F. In addition, the quality of the hatched chicks was assessed using the Pasgar©score. Following a 21-day incubation, chicks were obtained from DY1F and SY eggs. No chicks were obtained from DY2F eggs, although the embryos in these eggs developed up to the late stage of incubation. Early (≤7 d of incubation), middle (8–14 d), and late (≥15 d) embryonic mortality was significantly higher in DY eggs than in SY eggs. The embryonic mortality rate during early incubation was the same for DY1F and DY2F eggs, but middle and late embryonic mortality were significantly higher for DY2F eggs. Based on evaluation of embryo position according to Landauer, only three types of malposition that could potentially lead to embryonic death were noted. There were fewer malpositioned embryos in double-yolk eggs containing one embryo. Quality assessment of chicks (Pasgar©score) showed no differences between chicks hatched from eggs containing one yolk and those hatched from double-yolk eggs with one developing embryo, but chicks from double-yolk eggs were significantly heavier. The results of the research will contribute to a better understanding of the development and mortality of embryos in double-yolk eggs. Full article