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Avian metapneumovirus (aMPV) is a highly contagious pathogen that causes acute upper respiratory tract diseases in chickens and turkeys, resulting in serious economic losses. Subtype B aMPV has recently become the dominant epidemic strain in China. We developed an attenuated aMPV subtype B strain by serial passaging in Vero cells and evaluated its safety and efficacy as a vaccine candidate. The safety test showed that after the 30th passage, the LN16-A strain was fully attenuated, as clinical signs of infection and histological lesions were absent after inoculation. The LN16-A strain did not revert to a virulent strain after five serial passages in chickens. The genomic sequence of LN16-A differed from that of the parent wild-type LN16 (wtLN16) strain and had nine amino acid mutations. In chickens, a single immunization with LN16-A induced robust humoral and cellular immune responses, including the abundant production of neutralizing antibodies, CD4+ T lymphocytes, and the Th1 (IFN-γ) and Th2 (IL-4 and IL-6) cytokines. We also confirmed that LN16-A provided 100% protection against subtype B aMPV and significantly reduced viral shedding and turbinate inflammation. Our findings suggest that the LN16-A strain is a promising live attenuated vaccine candidate that can prevent infection with subtype B aMPV.

ABSTRACT: Live vaccines containing Eimeria oocysts are commercially available to protect against avian coccidiosis. Additionally, probiotics (PRO) and prebiotics (PRE) improve the poultry productivity and health and can be used as anticoccidial substitutes. However, the impact of PRO and PRE on reproductive potential, lesion score, intestinal health, and immunization outcomes of the live coccidia vaccines has not received adequate attention. Five groups of unsexed 1-day-old broiler chicks were used as follows: negative control (NC); challenged control (CC); vaccinated and challenged (VC); vaccinated, PRO-treated, and challenged (V-PRO); and vaccinated, PRE-treated, and challenged (V-PRE). At 21 d post-vaccination (pv), the vaccine increased the count of cecal anaerobes (P ≤ 0.05) and coliforms (P > 0.05) as well as harmed body weight gain (WG) (P ≤ 0.05), cecal lactic acid bacteria (P ≤ 0.05), and plasma carotenoid level (P > 0.05). None of the additives decreased oocyst shedding after vaccination, although they lowered the middle intestine and cecal lesion scores (P > 0.05). Compared to VC (2.68 ± 0.12) and V-PRE (2.66 ± 0.05), the V-PRO group showed an improved carotenoid level pv (2.96 ± 0.05) (P ≤ 0.05). V-PRE exhibited higher WG (822.95 ± 18.25) (P > 0.05) and FI (1153.01 ± 10.02) (P ≤ 0.05) than VC (781.86 ± 25.16 and 1109.85 ± 33.68) and V-PRO pv (787.61 ± 19.92 and 1077.43 ± 15.99). Following the homologous coccidia challenge, coccidia-vaccinated broilers adminstered the PRO or PRE continued to exhibit protection levels comparable to those received the vaccine alone. During 2 weeks post-challenge, VC, V-PRO and V-PRE improved bird performance and reduced oocyst shedding and lesion scores compared to CC. Ultimately, PRO and PRE treatments did not significantly reverse the reduction in growth performance in broiler chickens vaccinated against coccidia during the 1st three weeks of age.

ABSTRACT: Environmental conditions during incubation and rearing can influence stress responsivity of laying hen pullets throughout their lifespan, and therefore have important implications for their welfare. In this study, a 12:12h green LED light-dark cycle during incubation and larvae provisioning as enrichment during rearing were tested as strategies to optimize early-life conditions and thereby decrease stress responsivity in ISA Brown laying hens. A combination of parameters was measured to indicate neuronal, physiological, and behavioral changes that may affect fear and stress. The proteins calbindin D28k (calbindin1), doublecortin (DCX), and neuronal nuclein protein (NeuN) were quantified after hatch as a proxy for brain plasticity. Plasma and feather corticosterone levels were measured after hatch and at the end of the rearing phase, and fearfulness was investigated through a series of behavioral tests (i.e., voluntary approach, open field, tonic immobility, and manual restraint tests). No effects of light during incubation were found on calbindin1, DCX, or NeuN. Neither of the treatments affected corticosterone levels in blood plasma and feathers. Light-incubated pullets showed less fearfulness towards humans in the voluntary approach test, but not in the other behavioral tests reported in this study. Larvae provisioning had no effect on behavior. Our study showed minor effects of light during incubation and no effects of enrichment during rearing on stress responsivity of laying hen pullets. The small effects may be explained by the enriched rearing conditions for all birds in this experiment (low stocking density, natural daylight, and 24/7 classical music). Given the promising results of lighted incubation in other studies, which were mostly performed in broiler chickens, and evidence regarding the positive effects of enrichment during rearing, the potential of these strategies to improve laying hen welfare needs to be explored further.

ABSTRACT: Domestic laying hens rely primarily on their hindlimbs for terrestrial locomotion. Although they perform flapping flight, they appear to use maximal power during descent and thus may lack control for maneuvering and avoiding injuries on landing. This in turn may result in injury in open rearing systems. Wing-assisted incline running (WAIR) requires a bird to use its wings to assist the hindlimbs during climbing of an incline, and training in WAIR may therefore provide a useful method to increase a hen's power reserve and control for flight. We subjected hens to an exercise regimen involving inclines to induce WAIR for 16 wk during rearing. We then measured wing and body kinematics during aerial descent from a 155 cm platform. We hypothesized that birds reared with exercise would be better able to modulate their wing and body kinematics for making slower, more-controlled descent and landing. Brown-feathered birds exhibited greater wing beat frequencies than white-feathered birds, which is consistent with the higher wing loading of brown-feathered birds and WAIR-trained birds exhibited greater initial flight velocities compared to control birds. This may indicate that WAIR training provided an improved capacity to modulate flight velocity and strengthen the leg muscles. Providing incline exercises during rearing may therefore improve welfare for adult laying hens as greater initial flight velocity should reduce the power required for supporting body weight in the air and allow a hen to direct her excess power toward maneuvering.

Sperm motility, the percentage of sperm with forward progressions, is one of the semen quality traits that is highly related with male fertility potential in farm animals. The proportion of roosters with low sperm motility impedes the reproduction efficiency. This study aimed to determine crucial testicular proteins linked to sperm motility defects as causes of subfertility in roosters. iTRAQ was performed with the testis from five low sperm motility roosters and three high ones. A total of 2345 proteins were identified, of which, 86 were differentially expressed (DEPs), including 5 up-regulated and 88 down-regulated proteins in the low group. These DEPs were mainly enriched in cytoskeleton and cilium cell component, and biological processes related to sperm motility, and protein transport, and cellular protein localization. A cluster of 13 down-regulated proteins such as IFT88, TEKT1, ACTN2, DNAH5, RSPH9, and SPAG6, were associated with axoneme assembly. Further western blot and immunohistochemical analysis confirmed the down-regulated SPAG6 expression in low group, and indicated its expression in other cell types in testis beyond round spermatids, and that its pattern was in accordance with testis development and recession pace. Silencing transcription factor SOX5 down-regulated SPAG6 transcripts and impaired the cell proliferation and migration. In summary, this study highlights that down-regulated sperm flagellar structure associated proteins were the potential cause of low sperm motility. More specifically, the highly conserved SPAG6 protein across species is a positive regulator for testis development, spermatogenesis, and sperm motility regulation in chickens.

Homegrown eggs from free-ranging laying hens often contain elevated concentrations of perfluoroalkyl acids (PFAAs). However, it is unclear which factors contribute to these relatively large exposure risk scenarios. Moreover, existing bioavailability and modeling concepts of conventional organic pollutants cannot be generalized to PFAAs due to their different physicochemical soil interactions. Therefore, there is an urgent need for empirical models, based on real-world data, to provide insights into how (a)biotic factors affect the bioavailability to eggs. To this end, 17 targeted analytes were analyzed in abiotic (i.e. rainwater, soil; both N = 101) matrices and homegrown eggs (N = 101), which were sampled in 101 private gardens across Flanders (Belgium) in 2019, 2021 and 2022. Various soil characteristics were measured to evaluate their role in affecting PFAA bioavailability to the eggs. Finally, PFAAs were measured in potential feed sources (i.e. homegrown vegetable and earthworm pools; respectively N = 49 and N = 34) of the laying hens to evaluate their contribution to the egg burden. Modeling suggested that soil was a major exposure source to laying hens, accounting for 16–55% of the total variation in egg concentrations for dominant PFAAs. Moreover, concentrations in vegetables and earthworms for PFBA and PFOS, respectively, were significantly positively related with corresponding egg concentrations. Predictive models based on soil concentrations, total organic carbon (TOC), pH, clay content and exchangeable cations were successfully developed for major PFAAs, providing possibilities for time- and cost-effective risk assessment of PFAAs in homegrown eggs. Among other soil characteristics, TOC and clay content were related with lower and higher egg concentrations for most PFAAs, respectively. This suggests that bioavailability of PFAAs to the eggs is driven by complex physicochemical interactions of PFAAs with TOC and clay. Finally, remediation measures were formulated that are readily applicable to lower PFAA exposure via homegrown eggs.

Despite the continuous progress in food science and technology, the global burden of foodborne illnesses remains substantial, with pathogens in food causing millions of infections each year. Traditional microbiological culture methods are inadequate in detecting the full spectrum of these microorganisms, highlighting the need for more comprehensive detection strategies. This review paper aims to elucidate the relationship between foodborne pathogen colonization and the composition of the poultry microbiome, and how this knowledge can be used for improved food safety. Our review highlights that the relationship between pathogen colonization varies across different sections of the poultry microbiome. Further, our review suggests that the microbiome profile of poultry litter, farm soil, and farm dust may serve as potential indicators of the farm environment's food safety issues. We also agree that the microbiome of processed chicken samples may reveal potential pathogen contamination and food quality issues. In addition, utilizing predictive modeling techniques on the collected microbiome data, we suggest establishing correlations between particular taxonomic groups and the colonization of pathogens, thus providing insights into food safety, and offering a comprehensive overview of the microbial community. In conclusion, this review underscores the potential of microbiome analysis as a powerful tool in food safety, pathogen detection, and risk assessment.

ABSTRACT: Goose astrovirus genotype 2 (GAstV-2) is the causative agent causing severe visceral gout and joint gout in goslings, with mortality rates of affected flocks up to 50%. To date, continuous GAstV-2 outbreaks still pose a great threat to goose industry in China. Although most researches on GAstV-2 have focused on its pathogenicity to geese and ducks, limited studies have been performed on chickens. Herein, we inoculated 1-day-old specific pathogen-free (SPF) White Leghorn line chickens with 0.6 mL of GAstV-2 culture supernatant (TCID50 10−5.14/0.1 mL) via orally, subcutaneously and intramuscularly, and then assessed the pathogenicity. The results revealed that the infected chickens presented depression, anorexia, diarrhea, and weight loss. The infected chickens also suffered from extensive organ damage and had histopathological changes in the heart, liver, spleen, kidney, and thymuses. The infected chickens also had high viral load in tissues and shed virus after the challenge. Overall, our research demonstrates that GAstV-2 can infect chickens and adversely affect the productivity of animals. And the viruses shed by infected chickens can pose a potential risk to the same or other domestic landfowls.

This study aimed to investigate the nonclinical safety of lincomycin and spectinomycin hydrochloride (LC-SPH) intramuscular (i.m) doses on target animals (chickens) to provide guidelines for dose level design and side effect monitoring in clinical trials.A total of 80 healthy Arbor Acres plus broiler chicks were completely randomized and blindly divided into four treatment groups (control, one-time dose, three-time dose, and five-time dose) of 20 chicks each (20 chickens per group). At the age of day 15, all chickens (except the control group) were administered LC-SPH intramuscularly (chest muscles) at different doses of 20 mg/kg.bw, 60 mg/kg.bw, and 100 mg/kg.bw respectively for 9 consecutive days recommended by veterinary international cooperation on harmonization (VICH) guidelines. The chickens had ad libitum access to antibiotic-free feed and water. Feeding chickens were observed twice a day throughout the study. The drug safety was evaluated by complete blood count, biochemical parameters, histopathological, clinical signs, body weight gain, and feed conversion ratio (FCR).Hence, considering the minor toxicity of 60 mg/kg, our results reveal that intramuscular injection of at least 20 mg/kg body weight has no effects on growth performance, clinical blood parameters, organ coefficient, and histopathological parameters. Thus, a combination of LC-SPH 20 mg/kg body weight i.m injection investigated safe followed daily administration for nine consecutive days in healthy chickens.It is concluded that the experimental results support the safety of 20 mg/kg body weight in combination for the further clinical research study.

ABSTRACT: The genetic composition of populations is the result of a long-term process of selection and adaptation to specific environments and ecosystems. Runs of homozygosity (ROHs) are homozygous segments of the genome where the 2 haplotypes inherited from the parents are identical. The detection of ROH can be used to describe the genetic variability and quantify the level of inbreeding in an individual. Here, we investigated the occurrence and distribution of ROHs in 40 Beijing-You Chickens from the random breeding population (BJY_C) and 40 Beijing-You Chickens from the intramuscular fat (IMF) selection population (BJY_S). Principal component analysis (PCA) and maximum likelihood (ML) analyses showed that BJY_C was completely separated from the BJY_S. The nucleotide diversity of BJY_C was higher than that of BJY_S, and the decay rate of LD of BJY_C was faster. The ROHs were identified for a total of 7,101 in BJY_C and 9,273 in BJY_S, respectively. The ROH-based inbreeding estimate (FROH) of BJY_C was 0.079, which was significantly lower than that of BJY_S (FROH = 0.114). The results were the same as the estimates of the inbreeding coefficients calculated based on homozygosity (FHOM), the correlation between uniting gametes (FUNI), and the genomic relationship matrix (FGRM). Additionally, the distribution and number of ROH islands in chromosomes of BJY_C and BJY_S were significantly different. The ROH islands of BJY_S that included genes associated with lipid metabolism and fat deposition, such as CIDEA and S1PR1, were absent in BJY_C. However, GPR161 was detected in both populations, which is a candidate gene for the formation of the unique five-finger trait in Beijing-You chickens. Our findings contributed to the understanding of the genetic diversity of random or artificially selected populations, and allowed the accurate monitoring of population inbreeding using genomic information, as well as the detection of genomic regions that affect traits under selection.

SUMMARY: The objective of the present experiment was to investigate the influence of dietary crude protein (CP), standardized ileal digestible (SID) Lys level and their interaction in male Ross 308 broilers. From day 10, six experimental treatments were tested until day 35. The 6 experimental treatments consisted in a 3 × 2 factorial design with 3 levels of dietary SID Lys/CP (equivalent to 3 levels of dietary CP) and 2 levels of dietary SID Lys. Growth performance, uniformity, carcass traits, litter quality, blood & gut parameters, nitrogen (N) emissions, and global warming potential (GWP) were measured or calculated. There was no interaction for all criteria except for NH3 and N2O volatilization. The increase of dietary SID Lys/CP did not affect growth performance or uniformity, increased abdominal fat yield, decreased all N related parameters (litter N, blood uric acid, N excretion, NH3, and N2O volatilization). There was an interaction between dietary CP and AA origin for GWP; GWP was further reduced after CP reduction when using AA from EU. Increasing SID Lys improved growth performance, increased uniformity, increased breast meat yield, reduced abdominal fat yield, but increased N emissions and GWP. This study indicates that dietary CP can be drastically reduced in different nutritional contexts without negatively affecting growth performance, and with positive benefits on the environment.

ABSTRACT: Semen extenders are essential for maintaining the quality of sperm during storage and assuring the success of fertility after insemination. The objective of this study was to examine the effect of a new rooster semen extender (hereinafter, NCAB) supplemented with serine on the survivability and fertilizing ability of sperm following storage. NaCl solution, the NCAB extender, and IGGKPh extender were used as treatments. In Experiment 1, different storage temperatures (5°C and 25°C) and durations of storage (0, 12, and 24 h) were used to compare the semen quality and determine the suitable storage temperature. The fertility test was performed in in-station tests and on-farm experimental trials in Experiments 2 and 3, respectively. The results indicated that the interaction effect of duration, treatment, and storage temperature on all sperm parameters was highly significant. The NCAB extender significantly improved rooster semen quality during storage for 24 h at 5°C (P < 0.01). The sperm diluted in saline solution could not survive 24 h of storage at 25°C. The fertility and hatchability rates obtained for sperm diluted with the NCAB extender were higher than those diluted with other extenders. In addition, the fertilizing capacity of the NCAB extender-diluted sperm under field conditions was significantly higher than that of the saline solution-treated sperm. In conclusion, the NCAB extender supplemented with serine and stored at a low temperature (5°C) positively affects sperm quality and fertilization.

Black soldier fly larvae (BSFL) have garnered considerable attention for their efficacy in mitigating waste management challenges. However, their potential in treating antibiotics contaminated chicken manure remains uncertain. This study investigates the physicochemical properties changes and nutrient dynamics during the composting of contaminated-chicken manure using BSFL. The results indicate that BSFL treatment reduces electrical conductivity (by 6.01-58.09 %), organic matter, and dissolved organic carbon content in chicken manure throughout the composting process, while maintaining a more stable pH value (pH ∼ 6.0-8.0). This is attributed to the consumption of organic matter by BSFL and the subsequent promotion of organic acid formation. Additionally, BSFL treatment improves the degree of aromatization of dissolved organic matter (DOM) in chicken manure and increases the proportions of fulvic acid (up to 48.77 %) and humic acid (maximally 14.27 %) within the DOM. The germination index and pot experiments indicated improved compost maturity and plant growth in BSFL-treated composts. Furthermore, BSFL meal demonstrated high protein and essential fatty acid content, highlighting its potential as a protein supplement in animal feed. This study underscores the efficacy of BSFL in enhancing compost quality and nutrient availability, offering a sustainable solution for waste management and animal feed production., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Glyphosate (GLY) is the most universally used herbicide worldwide and its application has caused extensive pollution to the ecological environment. Increasing evidence has revealed the multi-organ toxicity of GLY in different species, but its male reproductive toxicity in avian species remains unknown. Thus, in vivo and in vitro studies were conducted to clarify this issue. Data firstly showed that chronic GLY exposure caused testicular pathological damage. Intriguingly, we identified and verified a marked down-regulation gap junction gene Connexin 43 (Cx43) in GLY-exposed rooster testis by transcriptome analysis. Cx43 generated by Sertoli cells acts as a key component of blood-testis barrier (BTB). To further investigate the cause of GLY-induced downregulation of Cx43 to disrupt BTB, we found that autophagy activation is revealed in GLY-exposed rooster testis and primary avian Sertoli cells. Moreover, GLY-induced Cx43 downregulation was significantly alleviated by ATG5 knockdown or CQ administration, respectively, demonstrating that GLY-induced autophagy activation contributed to Cx43 degradation. Mechanistically, GLY-induced autophagy activation and resultant Cx43 degradation was due to its direct interaction with ER-α. In summary, these findings demonstrate that chronic GLY exposure activates autophagy to induce Cx43 degradation, which causes BTB damage and resultant reproductive toxicity in roosters., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Microbial contamination annually leads to substantial food resource loss. Effective food packaging can mitigate food contamination and waste, yet conventional materials such as plastics often lack bacteriostatic activity. This study aimed to synthesise FengycinA-M3@bacterial cellulose@polyvinyl alcohol composite hydrogels via dual cross-linking with hydrogen and borate bonding, with the goal of enhancing antibacterial properties and prolonging the preservation period of refrigerated chicken breast. The composite hydrogel was subjected to comprehensive characterisation for structural, mechanical, water absorption, slow peptide release, antimicrobial capacity, biocompatibility, and chicken breast freshness preservation. The results showed that the composite hydrogel had a porous network structure and excellent gel elasticity and biocompatibility. It was effective in inhibiting Staphylococcus aureus and Escherichia coli, and prolonged the storage time of frozen chicken breast for up to 12 days. These findings emphasise the potential of hydrogel food packaging to prolong storage periods and its suitability for food industry applications due to ease of manufacture., Competing Interests: Declaration of competing interest The authors confirm that there are no conflicts of interest associated with this study., (Copyright © 2024 Elsevier B.V. All rights reserved.)

We aimed to create high-moisture plant-based meat alternatives (HPMAs) that closely mimic the fibrous structure found in conventional meat. To achieve this goal, we focused on the impact of modulating the heating and cooling die temperatures during the extrusion process on the structural properties of HPMAs. The optimal temperatures for the heating and cooling dies were determined to be 160 and 50 °C, respectively, following a comprehensive analysis of various quality characteristics. These included assessments of cutting strength, texturization degree, moisture content, and microstructure. Analyzing the correlation coefficients between heating and cooling die temperatures and the quality characteristics of HPMAs revealed that modulating the cooling die temperature has a greater impact on quality characteristics than modulating the heating die temperature. The quality characteristics of HPMAs were also compared with those of boiled and sous-vide cooked chicken breasts. The optimal HPMA exhibited the most similar fibrous macro- and micro-structure to sous-vide cooked chicken breast. Consequently, our study highlights the importance of modulating the heating die and cooling die temperatures, among various process parameters of high-moisture extrusion cooking, which is crucial for producing HPMAs closely resembling conventional meat., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Heat stress is a critical step in the processing of liquid egg white; however, this treatment can severely affect its foaming properties. This study aims to elucidate the mechanisms underlying the decline in foaming properties of liquid egg white during heat stress. The research begins by examining the adverse effects of heat stress on the foaming properties of liquid egg whites, where an increase in apparent viscosity, turbidity, and particle size is initially observed, indicating the formation of aggregates. After heat stress, the binding water capacity of the liquid egg white increases, intermolecular forces strengthen, and the secondary structure transforms towards β-sheet and β-turn configurations, while surface hydrophobicity decreases. Heat stress promotes the transition of liquid egg white into a more stable gel state. Additionally, electrophoresis results show the disappearance of bands for ovomucin subunit, ovotransferrin, and lysozyme, while microscopic observations reveal a rougher surface texture of the samples. In summary, this study provides insights and theoretical basis for understanding the mechanisms behind the decline in foaming properties of liquid egg whites under heat stress., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The molecular weight (MW) of oligosaccharides on gel properties of myofibrillar protein (MP) at high temperature remains unclear. In this study, it was found that chitosan oligosaccharides (CO) with different MW all significantly alleviated the textural deterioration of MP gel with high-temperature treatment. Moreover, MP-CO gel with the largest MW had the highest breaking force and the lowest cooking loss. Low-field NMR results further indicated that MP-CO gel with larger MW of CO had gradually increased relaxation rate, thus binding water more tightly. Rheological and microrheological tests suggested the addition of CO with larger MW resulted in much tighter gel network. These results indicated that CO with larger MW improved the quality of MP gel more effectively, which was because CO with larger MW inhibited aggregation of MP to a larger extent, resulting in smaller MP aggregates. Then MP-CO gel with much denser and more homogeneous structure was formed. Besides, MP-CO gel with larger MW of CO had higher content of β-sheet, resulting in MP gel with more ordered structure and better gel quality. Therefore, this study provided theoretical guidance for choosing the appropriate CO in improving texture of high temperature meat products., Competing Interests: Declaration of competing interest The authors declared that there were no competing financial interests or personal relationships that could influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study sought to develop an edible active film by integrating Pickering emulsions of forsythia essential oil and bacterial nanofibers at various concentrations into a film-forming matrix composed of funoran (F). The stability of the emulsions was evaluated through examination of the micro-morphology, particle size and distribution, 7-day emulsification index, and embedding rate of the Pickering emulsions. Subsequently, selected Pickering emulsions were incorporated into F to generate the edible active film. Scanning electron microscopy, Fourier-transform infrared spectroscopy, and Raman spectroscopy revealed that the Pickering emulsion was uniformly distributed throughout the F film, interconnected by hydrogen bonds. X-ray Diffraction spectra exhibited changes in peak intensity and shifts in position attributable to the edible active film. Pickering emulsion had a minimal impact on thermal stability. The film's tensile strength significantly increased, while elongation at break decreased. The heightened concentration of hydroxyl groups in the film led to increased thickness, reduced moisture content, and enhanced hygroscopicity. The edible active film exhibited superior antioxidant and antibacterial properties, thereby more efficiently shielding against oxygen and water vapour. In preservation tests involving chicken and lamb, the Pickering emulsion led to elevations in pH, total volatile basic nitrogen, and thiobarbituric acid reactive substance levels in the meat., Competing Interests: Declaration of competing interest All the authors declare no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Introduction: Fine microsurgical motor skill acquisition can be challenging. With increasing technological innovation, the methods of microsurgical skills acquisition may change. Studies show that laboratory-based microsurgical training programmes on a 2D microscope significantly improves the microsurgical skill acquisition of novices. However, it remains to be seen if these skills are transferable to a 3D microscope or if gaming agility is more important? We present a randomised control trial of three interventions, namely laboratory tabletop microscope training (LM), high-fidelity video gaming (Sony PlayStation 4 console; VG) and high-fidelity virtual reality gaming (Sony PlayStation VR console; VR) versus a control group., Methods: Forty novice medical students were block randomised to four groups: control (no intervention) n = 10, LM n = 10, VG n = 10 and VR n = 10. Participants performed chicken femoral artery anastomosis using the Aesculap Aeos® 3D microscope platform at the baseline and again after the intervention. Performance was evaluated using a modified structured assessment of microsurgery skills (mSAMS) score, time taken to complete anastomosis and time taken for suture placement by two blinded independent assessors., Result: No statistically significant difference was noted between the groups at the baseline. There was a statistically significant improvement in the LM arm between the baseline and post-training for mSAMS score and time for suture placement. In the VG, VR and control groups no statistically significant difference was observed., Conclusion: Our study demonstrates that during early microsurgical training, an intense laboratory-based microsurgical training programme significantly improves a novice's anastomotic performance on a 2D microscope, and these skills are transferable when a 3D anastomosis is carried out. However, focused gaming had no significant effect, and the results were akin to that of the non-intervention group., (Copyright © 2024 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.)

Heat shock protein 90 (HSP90) has a recognized anti-heat stress injury effect, but its function and corresponding molecular mechanism in heat-stressed hepatocytes are not fully understood, especially in tropical animals. In the present study, we identified several key factors affecting resistance to injury liver tissues from heat-stressed Wenchang chickens (a typical tropical species), such as HSP90, cellular pyroptosis and mitophagy. Heat stress upregulated the NLRP3/Caspase-1/GSDMD-N-mediated cellular pyroptosis pathway and the Pink1/Parkin-mediated mitophagy pathway in chicken hepatocytes, accompanied by the upregulation of HSP90. We also found that HSP90 overexpression significantly reduced heat stress-induced hepatocyte pyroptosis and enhanced mitophagy in primary hepatocytes from Wenchang chickens (PHWCs). HSP90 knockdown significantly increased heat stress-induced hepatocyte pyroptosis and decreased mitophagy in PHWCs. Interestingly, we performed immunoprecipitation and immunofluorescence colocalization and found that HSP90 and Pink1 can interact and directly regulate the level of mitophagy in PHWCs. Our results suggest that HSP90, which regulates Pink1, is an important factor in mitophagy that attenuates heat stress injury by inhibiting cellular pyroptosis., Competing Interests: Declaration of competing interest The author(s) declared no potential conflicts of interest with respect to the research, author- ship, and/or publication of this article., (Copyright © 2024. Published by Elsevier B.V.)

Background: Protein quality, evaluated using Digestible Indispensable Amino Acid Score (DIAAS), requires ileal digestibility values of individual indispensable amino acids (IAAs) in each protein. However, true tryptophan (Trp) digestibility has rarely been quantified in humans., Objective: To measure the true Trp digestibility and DIAAS of 2 H-intrinsically labeled plant and animal protein sources in humans, using the dual isotope tracer technique., Methods: The true Trp digestibility of 2 H intrinsically labeled plant proteins such as whole mung bean (n = 6) and dehulled mung bean (n = 6), chickpea (n = 5), and yellow pea (n = 5), and protein from animal source foods such as egg white (n = 6), whole egg (n = 6), chicken meat (n = 6), and goat milk (n = 7) was determined against the known digestibility of U- 13 C spirulina whole cell protein as reference, except for goat milk protein that was measured against free crystalline 13 C-Trp as reference. Banked samples from earlier studies conducted to determine true IAA digestibility of different protein sources were used for the analysis. DIAAS was calculated for each test protein using digestibility corrected IAA scores (mg IAA/g of protein) in comparison with the IAA requirement score for adults., Results: The true Trp digestibility of whole mung bean, dehulled mung bean, chickpea, yellow pea, egg white, whole egg, chicken meat, and goat milk were 67.6 ± 3.7%, 74.5 ± 4.4%, 72.6 ± 2.3%, 72.5 ± 2.2%, 89.7 ± 2.5%, 91.4 ± 2.6%, 95.9 ± 2.2%, and 92.8 ± 2.9%, respectively. The true Trp digestibility of plant protein sources was significantly lower than that of animal protein sources (P < 0.05). Trp was not a limiting IAA in all the tested proteins., Conclusion: The true Trp digestibility determined in this study ranged from 67.6 ± 3.7% to 95.9 ± 2.2% for whole mung bean and chicken meat, respectively, and adds to the database of individual true IAA digestibility of different protein sources., Trial Registration Number: This study was registered in Clinical Trials Registry of India (CTRI) with registration numbers CTRI/2017/11/010468, CTRI/2020/04/024512, and CTRI/2018/03/012265., Competing Interests: Conflict of interest The authors report no conflicts of interest., (Copyright © 2024 American Society for Nutrition. Published by Elsevier Inc. All rights reserved.)

E26-transforming specific (ETS) variant 6 (ETV6) is a transcription factor regulating the expression of interferon stimulating genes (ISGs) and involved in the embryonic development and hematopoietic regulation, but the role of ETV6 in host response to virus infection is not clear. In this study, we show that ETV6 was upregulated in DF-1 cells with poly(I:C) stimulation or IBDV, AIV and ARV infection via engagement of dsRNA by MDA5. Overexpression of ETV6 in DF-1 cells markedly inhibited IBDV-induced type I interferon (IFN-I) and ISGs expressions. In contrast, knockdown, or knockout of ETV6 remarkably inhibited IBDV replication via promoting IFN-I response. Furthermore, our data show that ETV6 negatively regulated host antiviral response to IBDV infection by interaction with TANK binding kinase 1 (TBK1) and subsequently inhibited its phosphorylation. These results uncovered a novel role of ETV6 as a pro-viral factor in host response by inhibiting TBK1 phosphorylation, furthering our understandings of RNA virus immunosuppression and providing a valuable clue to the development of antiviral reagents for the control of avian RNA virus infection., Competing Interests: Declaration of competing interest We declare there are no conflicts of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Environmental pollution poses a significant challenge to the poultry industry, leading to substantial losses and adverse effects on the health, production, and performance of avian species. In recent years, there has been growing interest in exploring natural compounds with potential protective effects against cadmium (Cd)-induced toxicity. Luteolin (LUT), a flavonoid found in various plants, has been studied for its antioxidant, anti-inflammatory, and cytoprotective properties. In this study, Su green shell grass chickens were divided into 4 groups: control, LUT (150 mg LUT), Cd (100 mg CdCl2), and Cd + LUT (100 mg CdCl2 + 150 mg LUT) groups for 1 month, respectively. The present study revealed that LUT maintained the morphology and functional activity of the liver and intestine. LUT alleviated Cd-induced impairment in the liver and intestinal biochemical indicators, suppressed Cd-induced liver fibrosis, mitigated liver and intestinal tissue damage. Additionally, LUT reduced oxidative stress and regulated the Cd-induced impairment in trace elements of the liver and intestine. Furthermore, LUT reduced Cd-induced liver inflammation, restored Cd-induced intestinal barrier function, and normalized Cd-induced serum proteins, including changes in the content of glutamyltranspeptidase. Moreover, LUT maintained Cd-induced disruption of gut microbiota and alleviated bacterial dysbiosis. Overall, these findings suggest that LUT holds promise as a potential therapeutic agent for mitigating the adverse effects of Cd-induced toxicity in poultry, by preserving liver and intestinal health, reducing oxidative stress, inflammation, and restoring gut microbiota balance., Competing Interests: DISCLOSURES The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Avian coccidiosis, caused by Eimeria spp, is a devastating disease in laying hens. Previous studies have suggested that amino acids may be involved in Eimeria infection of broiler chickens. However, their metabolic features in laying hens, as well as the effect of multiple Eimeria species challenges on poultry hosts have not been elucidated yet. Here, a targeted metabolomics approach was employed to identify altered amino acid metabolism and mechanisms in laying hens with multiple Eimeria species challenges. Laying hens, Hy-Line W-36 aged 25 wk, were randomly assigned to a control group and groups inoculated with varying levels of mixed Eimeria species (E. maxima, E. tenella, and E. acervulina). Serum samples from each group were collected at 6 d and 14 d of postinoculation (6 and 14 DPI) for metabolite profiling. Metabolomic analysis revealed notable metabolic variations between control and infected groups, especially at 6 DPI stage. Varying levels of Eimeria dosages did not show a significant metabolic difference, and metabolites were sensitive to low-level infection. With statistical analysis, differentially expressed compounds (3-methylhistidine, alanine, aspartate, lysine, asparagine, methionine, ornithine, and tryptophan) were selected, and their metabolic network was identified by pathway enrichment analysis. In the network, the lysine biosynthesis pathway was upregulated, while the arginine and proline metabolic pathway was downregulated under infection. Other pathways showed complex patterns of metabolic relationships. Based on the results, biological implications of metabolic changes were elucidated and discussed. Last, the results were further confirmed with our previous study (phenotype and gene expression results) using the same set of samples. Our finding provides in-depth information on altered amino acid metabolism and mechanisms in laying hens upon multiple Eimeria species infection., Competing Interests: DISCLOSURES The authors declare no conflict of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Valgus-varus deformity (VVD) is a common long bone deformity in broilers. Imbalance in cartilage homeostasis is the main feature of leg disease. Exosomes act as an important intercellular communication vector that regulates chondrogenesis by encapsulating specific nucleic acids and proteins. However, the exact mechanism of how plasma exosomal miRNAs regulate cartilage homeostasis in VVD broilers remains unclear. This study first demonstrated the structural disorder, growth retardation, and reduced proliferative capacity of VVD cartilage in vitro and in vivo. Subsequently, VVD and Normal broiler plasma exosomes were collected for miRNA sequencing. Cartilage-specific miR-455-5p was extraordinarily emphasized by performing bioinformatics analysis on differential miRNA target genes and further validated by tissue expression profiling. PKH67 fluorescently labeled plasma exosomes were shown to be taken up by chondrocytes, deliver miR-455-5p, inhibit chondrocyte proliferation, and disrupt their homeostasis, and these effects could be inhibited by the miR-inhibitors. Mechanistically, MiR-455-5p targets Ribosomal Protein S6 Kinase B1 (RPS6KB1) to inhibit RPS6 phosphorylation and reduce the synthesis of key proteins for cartilage proliferation, which in turn inhibits cartilage proliferation and disrupts its homeostasis. In conclusion, the present study identified abnormalities in VVD cartilage tissue and clarified the specific mechanism by which plasma exosome-derived miR-455-5p regulates cartilage homeostasis., Competing Interests: DISCLOSURES The authors declare no conflicts of interest., (Copyright © 2024. Published by Elsevier Inc.)

Zearalenone (ZEN), a mycotoxin produced by Fusarium species, is known for its reproductive toxicity as an estrogen analogue. However, there are limited knowledge about its hepatointestinal toxicity, as well as the role that gut microbiota and metabolites play in this process. In this study, a total of 24 thirty-week-old hens were fed to investigate the hepatointestinal toxicity subjected to long-term ZEN consumption at 2.0 mg/kg for 90 d. And we employed uncultured 16S rRNA sequencing for gut microbiota and untargeted metabolomics for fecal metabolites assessment. Notably, ZEN induced significant hepatic damage, as evidenced by hepatocyte necrosis, inflammatory cell infiltrate, increased liver lipopolysaccharide (LPS) and blood aspartate aminotransferase (AST) levels (P < 0.05). The decreased villus height, disruption of simple columnar epithelial cells, and exposure of the mucosal intrinsic layer were observed in the intestine. The gut microbial community composition and metabolites differed between ZEN group and control group. ZEN group exhibited higher gut microbial diversity (P < 0.05), lower Firmicutes/Bacteroidetes ratio and Lactobacillus abundance, and higher abundance in the genus such as Bacteroidetes, Parabacteroidetes and Desulfovibrio. Metabolomic analysis showed that ZEN treatment altered biosynthesis of siderophore group nonribosomal peptides and phenylpropanoids, metabolism of amino acid, digestion and absorption of vitamin and ABC transporters. Differential metabolites suggested that ZEN increase the risk of estrogen disorder, nucleic acid degradation, intestinal oxidative stress and inflammation. Neural network analysis showed that Ruminococcus was positively correlated with glyceric acid, and Prevotella was positively correlated with phenylacetylglycine. Both metabolites were positively correlated with blood AST level (P < 0.05), suggesting that intestinal microbe Ruminococcus and Prevotella might exacerbate liver damage by producing these harmful metabolites. Overall, we conclude that ZEN has damaged hepatointestinal system and the altered gut microbiota with resultant metabolite changes contribute to the adverse hepatointestinal effects of ZEN on laying hens. This study underscores the need for monitoring and mitigating ZEN exposure in poultry diets, highlighting its broader implications for animal health and food safety., Competing Interests: DISCLOSURES The authors declare no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Pathogenic bacteria infection, especially Clostridium perfringens (C. perfringens), markedly threatened the health of animals, and further caused huge economic loss. In this study, Bacillus licheniformis HJ0135 (BL) was used. Oxford cup bacteriostatic test and inhibitory rate test were conducted to evaluate the antibacterial ability of BL. Results showed the strongest inhibitory role of BL on C. perfringens (P < 0.05). Afterwards, 540 one-day-old yellow-feather broilers (32.7 ± 0.2 g) were randomly allocated into 3 groups, including CON group (basal diet), CP group (basal diet + 1 × 10 9 CFU C. perfringens in gavage), and BL + CP group (basal diet containing 7.5 × 10 6 CFU/g BL + 1 × 10 9 CFU C. perfringens in gavage). At d 70, broilers in the CP and BL + CP groups were treated with C. perfringens by continuously oral administration for 5 d. The experiment lasted for 75 d. The serum, immune organs, jejunal mucosa, and cecal contents were collected for analysis. In vivo experiment showed that BL supplementation markedly improved (P < 0.05) BW, ADG, thymus index, serum immunoglobins and antioxidases, reduced feed conversion ratio (FCR) and serum pro-inflammatory cytokines of C. perfringens-infected broilers. Furthermore, the increased jejunal injury and levels of pro-inflammatory cytokines, decreased gene expressions of tight junction proteins in the jejunal mucosa were significantly alleviated (P < 0.05) by BL. More importantly, the activation of NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3) inflammasome was inhibited (P < 0.05) by BL to further attenuate jejunal damage. Besides, BL supplementation markedly increased (P < 0.05) the cecal isobutyric acid and isovaleric acid. Microbial analysis showed that BL changed the composition and relative abundances of microbiota in the cecal contents (P < 0.05), especially the short chain fatty acids (SCFAs)-producing bacteria including Eubacterium&#95;coprostanoligenes&#95;group, Megamonas, Faecalibacterium, and Lactobacillus, which further protected against C. perfringens-induced jejunal inflammation in broilers. Our study laid a theoretical basis for the application of probiotics in lessening C. perfringens-related diseases in poultry farming., Competing Interests: DISCLOSURES The authors declare no conflicts of interest., (Copyright © 2024. Published by Elsevier Inc.)

The purpose of the study was to investigate the effects of the reduced dietary calcium (Ca) and phosphorus (P) level and supplementation of 25-hydroxycholecalciferol (25-OHD 3 ) on the expression of vitamin D receptor (VDR) and antimicrobial peptides and gut microbiota of broiler chickens with/without Eimeria challenge. A total of 576 fourteen-day-old broiler chicks were randomly allocated according to a 2 × 2 × 2 factorial design with main effects including Eimeria challenging (125,000 Eimeria acervulina, 25,000 Eimeria maxima, and 25,000 Eimeria tenella), dietary Ca and P levels (0.84% Ca and 0.42% available P or 0.64% Ca and 0.22% available P), and supplementation of 25-OHD 3 (3,000 IU/kg) of 6 replicates. Three-way ANOVA was performed, and the effects of 3 main factors and their interactions were investigated. The reduced dietary Ca and P level downregulated cathelicidins 3 (CATH3) in the upper jejunum in the Eimeria challenging condition (interaction; P < 0.05). The reduced dietary Ca and P level decreased the relative mRNA expression of jejunal avian beta defensin 5 (AvBD5) in the Eimeria challenging condition (interaction; P < 0.05). The reduced dietary Ca and P level tended to decrease the relative mRNA expression of jejunal AvBD9 in the Eimeria challenging condition (interaction; P = 0.051). The reduced dietary Ca and P level decreased observed features (alpha diversity parameter for richness) in the upper jejunal microbiota in the Eimeria challenging condition (interaction; P < 0.05). The supplementation of 25-OHD 3 decreased the relative abundance of the phylum Bacteroidetes (P < 0.05) and increased the relative abundance of the family Ruminococcaceae (P < 0.05) in the cecal digesta. The supplementation of 25-OHD 3 decreased the serum endotoxin level in the Eimeria challenging condition (interaction; P < 0.05). Therefore, the reduced dietary Ca and P level modulated the upper jejunal microbiota via modulating the expression of antimicrobial peptides, and the supplementation of 25-OHD 3 favorably modulated the cecal microbiota in broiler chickens with/without Eimeria challenge., Competing Interests: DISCLOSURES The authors declare no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The current study investigated the effects of dietary crude protein (CP) level on the liver lipid metabolism, gut microbiota, and bile acids (BA) profiles of growing pullets. Roman growing pullets (N = 180, 13-wk-old) were divided into 3 treatments groups with 6 replicates in each group and 10 hens in each replicate and provided 3 different dietary CP level diet treatments. The diet treatments included: a high-protein diet (15.5% CP, HP group), a medium-protein diet (14.5% CP, MP group), and a low-protein diet (13.5% CP, LP group). Compared with HP group, LP group significantly increased the lipid contents in the body (such as Breast intramuscular fat [BIMF], Leg intramuscular fat [LIMF], Percentage of abdominal fat [PAF], liver triglyceride [TG] and liver cholesterol [TC]), and the lipid metabolism-related parameters in serum (such as cholesterol (TC), high density lipoprotein cholesterol [HDL-C], low density lipoprotein cholesterol [LDL-C], very low density lipoprotein [VLDL]), and the mRNA expression of lipid metabolism-related genes (such as fatty acid synthase [FAS], CCAAT/enhancer binding protein β [C/EBPβ], and fatty acid translocase [FAT/CD6]) (P < 0.05). In addition, LP group significantly reduced the contents of lithocholic acid (LCA), isoLCA, and ursodesoxycholic acid (UDCA), and increased the deoxycholic acid (DCA) content compared with HP group (P < 0.05). The effects of LCA on lipid deposition were confirmed in chicken preadipocyte cell line (CPI), in which LCA supplementation significantly decreased the relative expression of PPARγ, FAS, acyl-CoA carboxylase (ACC) and SREBP-1c (P < 0.05). Correlation analysis further revealed a significant association between BA profiles and lipid metabolism-related parameters. Furthermore, 16S rRNA gene sequencing indicated that dietary protein level can significantly affect the richness, diversity, and composition of cecal microbiota in growing pullets. LP group significantly increased the abundance of Bacteroidetes and significantly decreased the abundance of Firmicutesa compared with the HP group. In summary, low protein diet in growing pullets influence the liver lipid metabolism through changing the gut microbiota and liver BA metabolism., Competing Interests: DISCLOSURES The authors declare no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

In poultry diets, copper is an essential nutrient that is critical for various physiological functions. Although copper sulfate is commonly used due to its cost-effectiveness, organic copper sources are gaining popularity because of their superior production outcomes and environmental benefits. Nevertheless, understanding the distinct bioaccessibility of inorganic and organic copper in diverse dietary setting remains limited. This study investigated the bioaccessibility of copper sulfate, copper amino acid chelate, and copper proteinate in the intestine via in vitro digestion and in situ dialysis. The results showed significant differences in the molecular size distribution of compounds formed by different copper salts within the intestinal environment, thereby leading to varying bioaccessibility. Copper sulfate has a bioaccessibility of 47 % ± 4%, which is significantly lower than copper amino acid chelate and copper proteinate (63% ± 5%, and 60% ± 4%, respectively) in purified diet systems. Similarly, in whey protein systems, sulfate records 54% ± 10% bioaccessibility compared to 78% ± 9% and 76% ± 5% for copper amino acid chelate and copper proteinate. Coexisting feed ingredients have a significant impact on copper bioaccessibility. Copper sulfate forms precipitates, reducing its bioaccessibility to 34% ± 1% in sodium nitrate solution. The addition of digestive enzyme increases the bioaccessibility of copper sulfate to 81% ± 2% by providing organic ligands. Digestive enzyme also enhanced the bioaccessibility of copper proteinate from 36% ± 4% to 81% ± 4% by degrading its ligands. However, feed ingredients may decrease copper bioaccessibility by forming macromolecular complexes with copper, as all the organic ligands can competitively bind with copper in the intestine. These findings emphasize the importance of considering copper salt types and diet composition in animal nutrition practices., Competing Interests: DISCLOSURES The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Resistance of Escherichia coli (E.coli) to antibiotics has steadily increased over time; hence, there is an urgent need to develop safer alternatives to antibiotics. The present study aimed to evaluate the effect of luteolin (Lut) on E. coli from chicken. The bioactive compound Lut from Humulus scandens was selected by network pharmacology and molecular docking analyses. Scanning electron microscopy (SEM), transmission electron microscopy (TEM), and confocal laser scanning microscopy (CLSM) were used to observe the effects of Lut on the morphology and structure of E. coli cells. The data-independent acquisition (DIA) method was used to analyze protein expression level of E. coli before and after Lut treatment. The in vivo evaluation of the antibacterial, anti-inflammatory, and oxidative effects of Lut on E.coli was conducted using E.coli isolated strains infected the SPF chicken model. The network pharmacology analysis revealed 19 distinctive bioactive compounds such as Lut and β-sitosterol in H. scandens; furthermore, 30 core targets were selected from H. scandens. The KEGG enrichment analysis showed that the PI3K-Akt, TNF, MAPK, IL-17, JAK-STAT, and HIF-1 pathways were related from H. scandens. Based on the results of the network pharmacology analysis, Lut was subjected to screening by molecular docking analysis to determine its antibacterial effect on E. coli and the associated mechanism of action. The minimum inhibitory concentration (MIC) of Lut against E. coli standard strains was 500 µg/mL. SEM, TEM, and CLSM results indicated that Lut damaged the cell wall and cell membrane of E. coli strains and destroyed the cell structure, leading to cell death.The expression level of membrane structure, Phenylalanine metabolism and some other metabolic pathways in E.coli changed after treatment with Lut (P < 0.05). In vivo experiments in the SPF chicken model showed that Lut treatment alleviated the decline in the growth performance of chickens (P < 0.05), prevented pathological changes in the correspond ding organs and suppressed the inflammatory response induced by E. coli infection (P < 0.05), improved the immunity and antioxidant capacity of chickens (P < 0.05), and protected them against infection with E. coli strains. To summarize, Lut from H. scandens can inhibit E. coli growth by damaging the cell membrane structureand affecting the expression level of some metabolic proteins. In vivo experiments also showed that Lut can significantly reduce the damage caused by E. coli isolates on SPF chickens, improve their antioxidant capacity and immunity, and reduce inflammatory responses following E. coli infection., Competing Interests: DISCLOSURES The authors declare that they have no conflict of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Glyphosate-based herbicides (GBH) are the most extensively used herbicides worldwide. Despite a presumed nondangerousness for animals, several studies reported negative effects after a GBH exposure in several animal models including birds, notably on reproductive functions. Several studies concerning the advantages of Vitamin E (VE) for antioxidant activity but also growth and reproduction have been reported in birds. However, it remains unclear whether VE could alleviate the negative effect of GBHs on chicken ovarian cells. Here we exposed chicken primary granulosa cells (GCs) from F1 and F3/4 follicles to growing doses of GBH (0.036, 0.36, 3.6, and 36 gly eq/L), with or without VE supplementation (1 mg/L) and investigated cell viability, proliferation, oxidative stress and steroidogenesis. GBH exposure did not affect F1 and F3 GCs viability but it increased cell proliferation only in F1 GCs and this effect was not altered by VE. In both F1 and F3/4 GCs, GBH exposure increased total oxidant status (TOS), reduced total antioxidant status (TAS) and consequently increased index of oxidative stress (OSI) in dose dependent manner. This latter effect for GBH 36 mg eq gly/L was totally abolished in response to VE. In both F1 and F3/4 GCs, GBH exposure reduced progesterone secretion in a dose dependent manner and this effect with GBH 0.36 and 1.8 mg eq glyphosate/L was alleviated by VE. However, we did not observe any effect of GBH and VE on the gene expression of several components of the steroidogenesis process. Taken together, these results show that GBH may have endocrine disruptor effects, and that these effects might be alleviated by antioxidant VE supplementation., Competing Interests: DISCLOSURES The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Enterococcus cecorum (EC) infection is an emerging endemic disease in UK and global broiler poultry with major economic impact and welfare concerns. There are significant research gaps with regards to EC pathogenesis, source of infection, transmission routes and early detection of disease, which this study aimed to address. In this prospective study, 725 environmental samples were collected from 4 broiler farms (A-D) the day before chick placement (d 1) and through the subsequent crop (d 7, 14, and 21). Cecal swabs were collected from birds that died of natural causes during the study period. A sample of birds that had been found dead or were culled for health reasons, were presented for post-mortem and samples were taken from lesions for EC culture. DNA was extracted from all environmental samples and EC detected using a qPCR and MALDI-TOF. Two EC isolates from diseased birds were inoculated on concrete slabs and incubated at 23°C and 32°C followed by swabbing of concrete culturing and determination of EC cfu at defined time points. Alongside environmental and bird sampling commercially available, smart camera systems were installed in selected houses on each farm to monitor bird activity and distribution. No EC outbreak occurred during the study, however, it was detected by qPCR in 215/725 (29.7 %) of all samples collected. Also, EC DNA was detected on average in 37% of samples collected on d 1, with approx. 88% of samples from chick paper being positive. Despite this, it was only cultured from 3 ceca samples and joint fluids of two infected birds from farm B on d 14 and 21. The survival experiments using isolates from infected chickens showed EC can survive on concrete for at least 21 d. This study provides invaluable insights into transmission pathways and tenacity of EC. Further studies are needed to determine strain characteristics in relation to their ability to cause disease and to further elucidate the sources of infection on poultry farms., Competing Interests: DISCLOSURES The authors declare no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Salmonella enteretidis (SE) has a great propensity to translocate from the cecum into internal organs such as the spleen and liver. However, a major concern is the ability of SE to colonize the ovaries. This study aimed to evaluate the efficacy of cell walls from Saccharomyces cerevisiae to control the Salmonella load in the ceca and ovaries of commercial layer pullets. Ten-week-old layer pullets were divided into 2 groups: one group was fed a control diet with commercial feed without additives, and another group was fed the same diet supplemented with 0.5 kg/metric ton of yeast cell walls (YCWs). At 16 wk of age, the birds in both groups were challenged with 3.0 × 10 9 CFU/mL SE by oral gavage. The birds were euthanized on d 7 and 14 postchallenge to collect the ceca and ovaries for Salmonella load determination. The results demonstrated that there were no statistical differences in ovary SE infection rates. The trend in the prevalence of SE positivity in the ovaries was similar at 14 d, with 2.1% (YCW pullets) to 4.2% positive for the ovaries of the nontreated pullets. There was also no significant difference in the SE log10 MPN/gram between the YCW and the control groups. In the ceca, the high level of SE (3.0 × 10 9 cfu/pullet), which results in ovarian transmission, causes high intestinal tract inflammation. There was a significant difference in the prevalence of SE in the ceca at 7 d postchallenge but not at 14 d postchallenge. In conclusion, the reduction in Salmonella load observed in the ceca on d 7 in this study shows the potential of YCW supplementation for reducing Salmonella colonization in poultry., Competing Interests: DISCLOSURES The authors declare no conflicts of interest. Declaration of Generative AI and AI-Assisted Technologies in the Writing Process: During the preparation of this work the author(s) do not used AI services., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The plasma membrane involves in many important biological events such as cell fusion and programmed cell death, but most of current plasma membrane probes cannot meet the requirement of long-term specific anchoring to the plasma membrane. Herein, we propose a molecular side-chain engineering strategy to modulate the long-term imaging performance of fluorescent dyes to the plasma membrane by regulating the cell permeability and anchoring ability. A series of FMR dyes with different lengths of side chains were designed and synthesized, and their transmembrane behaviours and staining performance were evaluated in living HeLa cells. We found that short-chain and medium-chain FMR dyes have excellent cell permeability without the labeling ability to the plasma membrane while the long-chain FMR dyes specifically stain the plasma membrane and can be firmly anchored to the plasma membrane for a long period of time. These long-chain FMR dyes have high stain specificality to the plasma membrane, and C10-FMR can be anchored to the plasma membrane of living cells for 2 h, which enables it to continuously monitor dynamic changes of the plasma membrane. The three-dimensional precision imaging of various cells was achieved using C10-FMR, which provides an opportunity to obtain complete information on the three-dimensional spatial morphology of the plasma membrane. The PEG-induced cell fusion of chicken red blood cells and H 2 O 2 -induced apoptosis of HeLa cells were monitored by real-time tracking of dynamic changes of the plasma membrane during these processes, which provide solid examples to prove the usefulness of these fluorescent dyes as long-term imaging tools. This work validates the hypothesis that cell permeability of membrane dyes can be readily regulated by tuning the side chains, and provides the effective design strategy of fluorescent dyes for 3D and long-term dynamic tracking of the plasma membrane of diverse animal cells., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Trimethyltin chloride (TMT) is widespread in the environment and is harmful to both humans and animals. In order to investigate the toxicity mechanism of TMT exposure on chicken liver, We established an in vivo experimental model by giving chickens oral administration of different concentrations of TMT dilution solution and vitro experiments of treating leghorn male hepatoma (LMH) cells for 12 h. The results showed that Albumin (ALB), total protein (TP) and alanine aminotransferase (ALT) in the blood of TMT-treated chickens, as well as ALT and aspartate aminotransferase (AST) in the liver, were dose-dependently increased, and different degrees of necrosis of hepatocytes were observed in histology. Meanwhile, TMT exposure led to a significant decrease in glutathione (GSH) content in chicken liver tissues and LMH cells, what's more a significant increase in malondialdehyde (MDA) content in cell supernatants. The expression of apoptosis-related genes Caspase8, Caspase3 and Caspase9 were increased in chicken liver tissues and LMH cells after treated by TMT, and an increased in the percentage of late apoptosis in LMH cells. This suggests that TMT can cause oxidative stress and apoptosis in chicken livers and cells, resulting in liver injury., Competing Interests: DISCLOSURES The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Cadmium (Cd) is a heavy metal that pollutes the environment and threatens human and animal health via the food chain. The spleen is one of the target organs affected by Cd toxicity. However, the mechanism of Cd toxicity is not fully understood. In this study, 80 chicks were allocated into 4 groups (n = 20) and exposed to different doses of CdCl 2 (0 mg/kg, 35 mg/kg, 70 mg/kg and 140 mg/kg) for 90 d. The pathological changes in the spleen, mitochondrial dynamics-related factors, cytochrome P450 (CYP450) enzyme system contents, activities, transcription levels, nuclear receptors (NRs) response molecule levels, and mitochondrial unfolded protein-related factors were detected. The findings indicate that exposure to Cd significantly leads to spleen injury. In Cd groups, the total contents of CYP450 and cytochrome b5 (Cyt b5) increased, and the activities of the CYP450 enzyme system (APND, ERND, AH, and NCR) changed. The NRs response was induced, and the gene levels of AHR/CAR and corresponding CYP450 isoforms (CYP1B1, CYP1A5, CYP1A1, CYP2C18, CYP2D6 and CYP3A4) were found altered. The study found that Cd exposure altered the mRNA expression levels of mitochondrial dynamics-related factors, such as OPA1, Fis1, MFF, Mfn1, and Mfn2, breaking mitochondrial fusion and cleavage and ultimately leading to mitochondrial dysfunction. Changes were detected in the gene levels of several mitochondrial unfolded protein response (mtUPR)-related factors, namely (SIRT1, PGC-1α, NRF1, TFAM, SOD2, and HtrA2). Cd also altered the gene levels of mitochondrial function-related factors (VDAC1, Cyt-C, COA6, PRDX3, RAF and SIRT3). It is showed that Cd can initiate the NRs response, influence the homeostasis of the CPY450 enzyme system, trigger the mtUPR, impair mitochondrial function, and ultimately lead to Cd toxicity in the spleen of chickens., Competing Interests: DISCLOSURES The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Most commercial laying hens suffer from sternum (keel) bone damage including deviations and fractures. X-raying hens, followed by segmenting and assessing the keel bone, is a key to automating the monitoring of keel bone condition. The aim of the current work is to train a deep learning model to segment the keel bone out of whole-body x-ray images. We obtained full-body x-ray images of laying hens (n = 1,051) and manually drew the outline of the keel bone on each image. Using the annotated images, a U-net model was then trained to segment the keel bone. The proposed model was evaluated using 5-fold cross validation. We obtained high segmentation accuracy (Dice coefficients of 0.88-0.90) repeatably over several validation folds. In conclusion, automatic segmentation of the keel bone from full-body x-ray images is possible with good accuracy. Segmentation is a requirement for automated measurements of keel geometry and density, which can subsequently be connected to susceptibility to keel deviations and fractures., Competing Interests: DISCLOSURES The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Fatty liver disease in laying hens, characterized by excessive lipid accumulation in hepatocytes, poses significant challenges to poultry health and production efficiency. In this study, we investigated the therapeutic potential of epigallocatechin gallate (EGCG), a bioactive compound found in green tea, in mitigating oleic acid (OA)-induced hepatic steatosis in primary chicken hepatocytes. Treatment with EGCG effectively attenuated lipid deposition by downregulating lipid synthesis-related genes. Moreover, EGCG mitigated oxidative stress, inflammation, DNA damage, and apoptosis induced by OA, thereby preserving hepatocyte viability. Mechanistically, EGCG exerted its protective effects by modulating the p38 MAPK signaling pathway. Our findings suggest that EGCG holds promise as a therapeutic agent for managing fatty liver disease in poultry, offering insights into novel strategies for improving poultry health and production outcomes., Competing Interests: DISCLOSURES The authors declare that they have no known competing financial interests., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Salmonellosis is still one of the most reported zoonoses worldwide and poultry meat is a major source, as chickens are often persistent carriers of Salmonella. Medium-chain fatty acids (MCFA) are known for their strong antimicrobial activity. MCFAs used today in the animal feed industry, however, mainly originate from the palm oil industry, which is notorious for its negative impact on the climate. We investigated the effect of a specific blend of palm-free MCFAs (ranging from C6 to C9) on Salmonella Enteritidis (SE) colonization in broiler chickens and in vitro SE characteristics. Fifty Ross 308 broiler chickens were randomly divided in 2 treatment groups. Chickens received either un-supplemented feed or feed supplemented with 300 ppm MCFAs from D0 onwards. On D7, all chickens were orally inoculated with 1600 CFU of SE. Cloacal swabs (D11) and samples of liver and caeca (D12) of all animals were collected and SE was enumerated. Percentage of SE-positive caecum samples was significantly (P = 0.044) reduced in birds receiving MCFAs compared to those receiving unsupplemented feed (36% vs. 64%). In vitro work performed with the same SE strain showed that preincubating the Salmonella bacteria with MCFAs at a sub-minimal inhibitory concentration significantly (p < 0.05) reduced bacterial adhesion to and invasion in Caco-2 cells, which may explain the observed reduction in intestinal SE colonization in the in vivo trial. Together, these results show that the tested eco-friendly MCFA blend could be a promising tool in the control of Salmonella in broilers., Competing Interests: DISCLOSURES The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

To explore the potential protective effect of resveratrol (RES) on cold-exposed broilers, 360 21-day-old broilers were equally divided into 5 groups with 6 replicates. A control (CON) group was reared at the normal feeding temperature and received a basal diet, and 4 cold exposure (8 ± 1°C for 10 h/d from d 29 to 42) groups were fed the basal diet with 0 (CE), 250 (CE + RES250), 500 (CE + RES500), and 750 (CE + RES750) mg/kg RES from d 22 to 42. Broilers were slaughtered on d 42 and heart tissues were collected to measure the relevant indexes. The results showed that heart tissues of all CE-broilers had inflammatory cell infiltrations, and dietary RES supplementation reduced this phenomenon. Compared to CON group, the concentrations of MDA and H 2 O 2 were increased and activities of SOD and CAT were decreased in all CE-broilers (P < 0.05). mRNA expression of genes related to endoplasmic reticulum (ER) stress (GRP78, IRE1, PERK, EIF-2α, ATF4, ATF6, and CHOP), pyroptosis (NLRP3, ASC, Caspase1, GSDME, IL-18, and IL-1β), and proinflammation (TNF-α, IFN-γ, IL-2, and IL-6) was upregulated and that of ant-inflammatory cytokines (IL-4 and IL-10) was downregulated in CE and all CE + RES groups compared to CON group (P < 0.05). Compared to CE group, the activities of SOD and CAT and mRNA expression of anti-inflammatory genes were increased (P < 0.05), and concentrations of MDA and H 2 O 2 and mRNA expression of ER stress, pyroptosis and proinflammatory genes were reduced (P < 0.05) in 3 CE + RES groups. Additionally, protein levels of PERK, ATF4, CHOP, NLRP3, Caspase1, GSDMD, IL-18, IL-1β, TNF-α, and IL-10 were similar in their mRNA expression. Overall, cold exposure caused oxidative stress and ER stress, and induced pyroptosis and inflammatory response, resulting in heart injury in broilers, and dietary RES addition reduced heart damage by enhancing antioxidant defense function. This study indicates that RES can be a feed additive to alleviate cold exposure-induced heart injury in broilers, and a 500 mg RES/kg diet is the optimal supplemental level., Competing Interests: DISCLOSURES The authors confirm that there are no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The clinical relevance of avian metapneumovirus (aMPV) is growing in the poultry sector, especially in broiler farming, where no vaccination is administered in Italy. Given the naïve status of the birds, a serological survey was conducted in a densely populated area of Northern Italy, to evaluate aMPV circulation. Seven farms were selected and sampled in summer/fall, then sampling was repeated in the following season (winter/spring) to assess a possible seasonal effect. In each farm, fifteen birds were blood sampled towards the end of the cycle and sera were analyzed with an ELISA test. Clinical signs were reported in 5 out of 7 farms, although all farms were positive at both sampling points, except for one, which was negative at the first sampling. The seroprevalence within farm ranged from 26.6% to 100%, and antibody titres appear to increase with age. No seasonality effect was evidenced, whereas a farm effect was more distinct. aMPV circulation appears wide in Northern Italian farms, with different clinical outcomes that could be modulated by intrinsic characteristics of the farms. In absence of vaccination, serological monitoring can be a useful tool for viral entrance monitoring, although sampling timing should be evaluated in order to spot seroconversion after late infections., Competing Interests: DISCLOSURES The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Avian reovirus (ARV) is the causative agent of avian viral arthritis and causes significant economic losses to the global poultry industry. For clinical diagnosis, detecting ARV-specific antibodies is crucial. We successfully expressed the ARV-σC protein in insect cells using the baculovirus expression vector system, achieving an expression level of approximately 200 mg/L. We developed an indirect enzyme-linked immunosorbent assay (iELISA) using the ARV-σC protein as a coating antigen to detect antibodies against it. The inter-batch and intrabatch coefficients of iELISA variation were less than 10%. Its sensitivity (1:12,800 diluted in serum) was 4 times higher than that of the indirect immunofluorescence assay (IFA; 1:3200 diluted in serum), and it showed no cross-reactivity with antibodies against other common avian viruses (such as Infectious bursal disease virus, Newcastle disease virus). The practicality of the iELISA was further evaluated using clinical samples. 300 clinical sera from chickens vaccinated with the ARV attenuated vaccine and 20 SPF sera were tested using both the iELISA and the IFA, demonstrating a 100% conformity rate. In conclusion, these results suggest that the iELISA developed in this study is a rapid, sensitive, and specific method that could serve as an effective diagnostic tool for monitoring and controlling avian viral arthritis., Competing Interests: DISCLOSURES All authors disclosed no relevant relationships., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Gyrovirus galga1 (GyVg1), formerly known as AGV2, was initially identified in chickens in southern Brazil. The prevalence of GyVg1 from 2021 to 2024 in 28 out of the 63 poultry farms located in Jiangsu, Anhui, Henan, Hunan, Shandong, and Hubei provinces in eastern and central China was detected via PCR. The complete genomes of the 28 strains were sequenced and exhibited a full length of 2,376 bp. Similarity analysis of these strains did not suggest definite correlation with evolutionary branching and geographical distribution. Compared with the reference GyVg1 strains, HN2202 shared the highest similarity of 99.71% with HLJ1511 (chicken-originated) from northeastern China in 2015 to 2016. Recombination analysis revealed that AH2102 was a potential recombinant of peafowl-originated HN2019-PF1 and chicken-originated HLJ1506-2, whereas HN2304 was a recombinant of peafowl-originated HN2019-PF1 and the Hungarian ferret strain G13. Mutation site analysis of the capsid protein revealed that highly mutated regions occurred between sites 288 to 316 and 383 to 419. These results indicate that GyVg1 may have undergone an interspecies transmission, which involved complex mutations and recombination. This study may provide a reference for subsequent investigations targeting the molecular epidemiology and viral evolution of GyVg1., Competing Interests: DISCLOSURES The authors have no competing interests to declare., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The hyperplasia and hypertrophy of preadipocytes were closely related to lipid deposition in animals. Butyric acid was reported to be involved in lipid metabolism. The aim of the current study was to investigate the effect of butyric acid on the proliferation and differentiation of the immortalized chicken preadipocyte 2 (ICP2). ICP2 were treated respectively with 12mM butyric acid for 48h in proliferation trial and 4mM butyric acid plus 200 μM oleic acid for 3 d in differentiation trial. For the proliferation trial, RNA-seq analysis revealed that 2039 genes were significantly up-regulated and 780 genes were significantly down-regulated with 12 mM butyric acid after 48 h treatment. Concurrently, Cell cycle, DNA replication and p53 signaling pathways were down-regulated in Butyric acid group. More importantly, 12 mM butyric acid restrained the expression of cell proliferation genes such as PCNA, CDK1 and CDK2 in Butyric acid group (P < 0.05), and the protein expression levels of PCNA and CDK1 were also significantly decreased (P < 0.05). The Oil red staining revealed a fewer presence of red fat droplets in ICP2 following treatment with 4 mM butyric acid, accompanied by decreased levels of total cholesterol (TC) and triglycerides (TG). RNA-seq analysis shown that the number of up and down-regulated genes were 2095 and 1042 respectively in OAB group (oleic acid+butyric acid) when compared with OA group (oleic acid). Meanwhile the AMPK signaling pathway, FOXO signaling pathway and focal adhesion were significantly enriched in OAB group. Additionally, 4 mM butyric acid inhibited the expression of lipid differentiation genes including FABP4, C/EBPα, PPARγ and LPL in OAB group (P < 0.05), as well as lipogenesis proteins such as FABP4, C/EBP-α and PPARγ (P < 0.05). In conclusion, 12 mM butyric acid effectively inhibited the proliferation of ICP2 by slowing down cell cycle progression, while 4 mM butyric acid alleviated lipid deposition by reducing the production of lipid droplets through inhibiting the expression of lipid differentiation marker genes and proteins., Competing Interests: DISCLOSURES The author affirmed that all data in this study are freely accessible without limitation, and declared that they have no competing financial interests or personal relationships in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

As the main active glycoprotein of egg white, the biological functions of chicken ovomucin α- and β-subunit are closely related to the structure of glycans. However, the exact composition and structure of the subunit glycans are still unknown. We obtained highly pure chicken ovomucin α-subunit and β-subunit protein bands by the strategy combined with two-step isoelectric precipitation and SDS-PAGE gel electrophoresis. The ammonia-catalyzed one-pot procedure was then used to release and capture α-and β-subunit protein glycans with 1-phenyl- 3-Methyl-5-pyrazolone (PMP). The N/O-glycans of bis-PMP derivatives were purified and analyzed by LC-MS. More importantly, an effective dual modification was performed to accurately quantify neutral and sialylated O-glycans through methylamidation of sialic acid residues and simultaneously through carbonyl condensation reactions of reducing ends with PMP. We first showed that the α-subunit protein has only N-glycosylation modification, and the β-subunit only O-glycosylation, a total of 22 N-glycans and 20 O-glycans were identified in the α- and β-subunit, respectively. In addition, the complex N-glycan (47 %) and the sialylated O-glycan (77 %) are each major types of the above subunits. Such findings in this study provide a basis for studying the functional and biological activities of chicken ovomucin glycans., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Improper disposal of organic solid waste results in serious environmental pollution. Aerobic composting provides an environmentally friendly treatment method, but improving humification of raw materials remains a challenge. This study revealed the effect of different concentrations of potassium persulfate (PP) on humification of chicken manure and straw aerobic composting and the underlying microbial mechanisms. The results showed that when 0.6 % PP was added (PPH group), humus and the degree of polymerization were 80.77 mg/g and 2.52, respectively, which were significantly higher than those in 0.3 % PP (PPL group). As the concentration of PP was increased, the composition of rare taxa (RT) changed and improved in evenness, while abundant taxa (AT) was unaffected. Additionally, the density (0.037), edges (3278), and average degree (15.21) in the co-occurrence network decreased compared to PPL, while the average path (4.021) and modularity increased in PPH. This resulted in facilitating the turnover of matter, information, and energy among the microbes. Interestingly, cooperative behavior between microorganisms during the maturation period (24-60 d) occurred in PPH, but competitive relationships dominated in PPL. Cooperative behavior was positively correlated with humus (p < 0.05). Because the indices, such as higher degree, betweenness centrality, eigenvector centrality, and closeness centrality of the AT, were located in the microbial network center compared to RT, they were unaffected by the concentration of PP. The abundance of carbohydrate and amino acid metabolic pathways, which play an important role in humification, were higher in PPH. These findings contribute to understanding the relative importance of composition, interactions, and metabolic functionality of RT and AT on humification during chicken manure and straw aerobic composting under different concentrations of PP, as well as provide a basic reference for use of various conditioning agents to promote humification of organic solid waste., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Avian primordial germ cells (PGCs) are important culture cells for the production of transgenic chickens and preservation of the genetic resources of endangered species; however, culturing these cells in vitro proves challenging. Although the proliferation of chicken PGCs is dependent on insulin, the underlying molecular mechanisms remain unclear. In the present study, we explored the expression of the PI3K/AKT signaling pathway in PGCs, investigated its effects on PGC self-renewal and biological properties, and identified the underlying mechanisms. Our findings indicated that although supplementation with the PI3K/AKT activator IGF-1 failed to promote proliferation under the assessed culture conditions, the PI3K/AKT inhibitor LY294002 resulted in retarded cell proliferation and reduced expression of germ cell-related markers. We further demonstrated that inhibition of PI3K/AKT regulates the cell cycle and promotes apoptosis in PGCs by activating the expression of BAX and inhibiting that of Bcl-2. These findings indicated that the PI3K/AKT pathway is required for cell renewal, apoptosis, and maintenance of the reproductive potential in chicken PGCs. This study aimed to provide a theoretical basis for the optimization and improvement of a culture system for chicken PGCs and provide insights into the self-renewal of vertebrate PGCs as well as potential evolutionary changes in this unique cell population., Competing Interests: DISCLOSURES The authors declare no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Naringenin is a flavonoid with significant anti-inflammatory and antioxidant properties. Mitochondrial dynamics, the mitochondrial respiratory chain, and mtROS are closely related to each other and regulate various biological processes. Ferroptosis is closely related to inflammatory responses and immune function in multiple tissues and organs. However, whether naringenin can alleviate LPS-induced inflammation and immune disorders in the chicken thymus via mtROS/ferroptosis has not been reported. Therefore, in this study, we constructed chicken thymus and MSB-1 cell models of LPS and naringenin based on screening for naringenin concentrations that have positive effects on inflammation and immune function to further investigate the anti-inflammatory, antiferroptosis, and maintenance of the immune function of naringenin. The results showed that 40 mg/kg naringenin alleviated LPS-induced tissue damage, elevated serum inflammatory factors, and decreased serum immune factors. The mechanism by which naringenin attenuates mtROS release by alleviating the imbalance of mitochondrial dynamics and the blockage of the respiratory chain. The effect of naringenin on alleviating LPS-induced lipid peroxidation, disruption of the GSH/GSSG system, iron overload, and GPx4 inactivation, thereby attenuating ferroptosis in thymus tissue, was inhibited by the addition of mtROS activators. In conclusion, naringenin alleviates LPS-induced ferroptosis in chicken thymus by attenuating mtROS release., Competing Interests: DISCLOSURES The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)