Search Results (265)

The collection of perivitelline fluid (PVF) from early-stage post-activation zebrafish (Danio rerio) eggs/embryos poses a significant challenge owing to the liability of the egg/embryo to sustain damage and rupture during handling. Rupture of the blastoderm and/or yolk presents a major risk of PVF sample contamination. Previous efforts to extract PVF at such early stages have employed formalin fixation to enhance the structural integrity of the blastoderm and yolk syncytial layer, thereby reducing the likelihood of contamination. While this approach successfully mitigates blastoderm and yolk rupture, formaldehyde fixation may cause issues with downstream proteomic analyses. Recent findings indicate that zebrafish PVF contains a range of maternally inherited proteins involved in innate immune defence. However, current extraction methods compromise the reliability of downstream protein analyses, raising concerns that fixation-induced protein crosslinking may obscure the presence of maternally inherited proteins during the earliest stages of development. The micro-aspiration technique described here allows for the precise extraction of PVF from living, water-activated eggs with minimal disruption to the blastodisc and yolk. This method reduces the risk of contamination from other non-target proteinaceous egg sources and eliminates the need for formalin fixation, thereby improving the integrity of PVF samples and enhancing the reliability of subsequent downstream analyses. Full article

Background: The metabolic dysfunction-associated steatotic liver disease (MASLD) represents an escalating global health concern, with effective treatments still lacking. Given its complex pathogenesis, multi-targeted strategies are highly desirable. Methods: This study reports the isolation of four flavone C-glycosides (FCGs) from Dianthus superbus L. and explores their potential in treating MASLD. The bioactivity and underlying mechanisms of FCGs were systematically evaluated by integrating network pharmacology, molecular docking, and zebrafish model validation. Results: Network pharmacology analysis revealed that FCGs may modulate multiple MASLD-related pathways, including lipid metabolism, insulin signaling, inflammation, and apoptosis. Molecular docking further confirmed strong binding affinities between FCGs and key protein targets involved in these pathways. In the zebrafish model of MASLD induced by egg yolk powder, FCGs administration markedly attenuated obesity, hepatic lipid accumulation, and liver tissue damage. Furthermore, FCGs improved lipid metabolism and restored locomotor function. Molecular analyses confirmed that FCGs upregulated PPARγ expression to promote lipid metabolism, restored insulin signaling by enhancing INSR, PI3K, and AKT expression, and suppressed inflammation by downregulating TNF, IL-6 and NF-κB. Additionally, FCGs inhibited hepatocyte apoptosis by elevating the BCL-2/BAX ratio. Conclusions: These findings highlight the multi-pathway regulatory effects of FCGs in MASLD, underscoring its potential as a novel therapeutic candidate for further preclinical development. Full article

The objective of this study was to develop a novel sauce formulation in which egg yolk was substituted with pea and soy proteins, in addition to the incorporation of tomato pomace as a functional ingredient. Nine experimental samples (E1–E3, S1–S3, and P1–P3) and three control samples (E0, S0, and P0) were prepared, corresponding to three protein sources (E: egg yolk, S: soy, P: pea), with increasing concentrations of tomato pomace (0, 2, 4, and 6%). The formulations were adjusted proportionally in terms of water and oil to maintain the desired consistency. The analyses performed included: physico-chemical analysis of the sauce (fat content, peroxide value, and CIE L* a* b* color determination), quality assessment using Fourier Transform Infrared Spectroscopy (FT-IR, rheological measurements, and microstructural evaluation. The sample designated P2 demonstrated a notable correlation with favourable parameters, exhibiting intense colouration, elevated protein content, and consistent rheological properties. However, at higher levels of tomato pomace (notably 6%), microstructural instability was observed, which may limit the formulation’s robustness over time. These findings demonstrate that tomato pomace can enhance the functional and structural characteristics of sauce, while also highlighting the importance of optimizing concentration levels to avoid negative impacts on emulsion stability. Overall, the results support the use of tomato pomace and plant proteins in the formulation of sustainable and innovative food products. Full article

The objective of this study was to evaluate the effects of various feed additives on odor emissions, gut health, and stress responses in laying hens fed low-protein diets. Four commercially available functional feed additives (Bacillus subtilis, protease, saponin, and thyme-based essential oil) were selected for this study. A total of 288 Hy-Line brown laying hens aged 49 weeks were randomly fed on one of six experiment diets: a 16% standard crude protein diet, a 12% low-crude-protein (LCP) diet, and LCP diets supplemented with Bacillus-based probiotic, protease, saponin, or thyme-based essential oils prepared for 8 weeks. Each treatment had eight replicates with six birds per replicate. Lowering crude protein levels affected the laying performance, nitrogen balance, odor production (i.e., ammonia), and nutrient digestibility but did not alter eggshell quality or fecal short-chain fatty acids. Dietary additives added into the LCP diet did not affect the laying performance, egg qualities, and nitrogen balance but increased crude ash digestibility compared with the LCP-diet-fed laying hens. Branched-chain fatty acids tended to be higher in all laying hens fed low-CP diets, irrespective of feed additives. Notably, low vs. standard protein diets tended to increase yolk corticosterone levels, which is an indicator of stress responses in chickens. This low-CP-mediated increase in yolk corticosterone was partially decreased by 20.8–48.6% on average, depending on the additives used. Our study suggests that low-protein diets could effectively lower nitrogen excretion and odor emissions. However, adding dietary additives into low-protein diets has minimal effects on low-CP-diet-fed laying hens, which needs further studies to clarify the role of low-crude-protein diets and dietary additives in modulating hindgut fermentation via shaping the gut microbiota and stress responses of laying hens. Full article

Cytoplasmic polyadenylation element-binding proteins (CPEBs) are critical regulators of maternal mRNA translation during oogenesis, yet their roles in insect reproduction remain underexplored. Here, we characterized CmCPEB2, a CPEB homolog in the rice leaf roller Cnaphalocrocis medinalis, a destructive lepidopteran pest insect, and elucidated its role in mating-induced oviposition. The CmCPEB2 protein harbored conserved RNA recognition motifs and a ZZ-type zinc finger domain and was phylogenetically clustered with lepidopteran orthologs. Spatiotemporal expression profiling revealed CmCPEB2 was predominantly expressed in ovaries post-mating, peaking at 12 h with a 6.75-fold increase in transcript levels. Liposome-mediated RNA interference targeting CmCPEB2 resulted in a 52% reduction in transcript abundance, leading to significant defects in ovarian maturation, diminished vitellogenin deposition, and a 36.7% decline in fecundity. The transcriptomic analysis of RNAi-treated ovaries identified 512 differentially expressed genes, with downregulated genes enriched in chorion formation and epithelial cell development. Tissue culture-based hormonal assays demonstrated the juvenile hormone-dependent regulation of CmCPEB2, as JH treatment induced its transcription, while knockdown of the JH-responsive transcription factor CmKr-h1 in the moths suppressed CmCPEB2 expression post-mating. These findings established CmCPEB2 as a juvenile hormone-dependent regulator of mating-induced oviposition that orchestrates vitellogenesis through yolk protein synthesis and ovarian deposition and choriogenesis via transcriptional control of chorion-related genes. This study provides novel evidence of CPEB2-mediated reproductive regulation in Lepidoptera, highlighting its dual role in nutrient allocation and structural eggshell formation during insect oogenesis and oviposition. Full article

This study aimed to develop a shelf-stable, plant-based whole hard-boiled egg analogue, available in both regular and low-allergenic versions. Six plant proteins—soy, mung bean, pea, rice, potato, and wheat—were formulated into egg white and yolk components, with mung bean and wheat proteins showing the most promising sensory and visual qualities. Two preservation methods, thermal pasteurization (75–85 °C, 15–20 min) and gamma irradiation (2–5 kGy), were applied to extend shelf life while maintaining product quality. Thermal treatment at 75 °C for 15 min and gamma irradiation at 3.5 kGy were identified as optimal conditions, balancing sensory acceptability and microbial safety. Sensory evaluation by 100 untrained panelists revealed favorable scores for appearance, texture, and overall liking, without significant differences among selected formulations (p > 0.05). Accelerated shelf life testing and Q10 modeling predicted a shelf life of 188 days for thermally pasteurized eggs and 253 days (8.42 months) for gamma-irradiated eggs at 30 °C. These results demonstrate the feasibility of developing a consumer-acceptable, plant-based hard-boiled egg analogue with extended ambient shelf life. Full article

Aspects related to the contamination of hatching eggs, sanitary management during pre-incubation, and the performance of the incubation process can compromise productive efficiency in poultry farming. When these factors negatively influence poultry farming, they can destabilize the generation and distribution of financial resources throughout the production chain, as well as limit public access to poultry-derived proteins. Understanding how these aspects are interrelated is essential for making decisions that benefit poultry health and productivity. Therefore, we conducted a multivariate analysis of microbiological and incubation parameters to evaluate whether bacterial contamination of the eggshell and yolk sac negatively affects HI and to compare the effectiveness of different sanitization protocols in reducing bacterial contamination in these regions. To achieve this, we utilized the raw data from our previous research on the sanitization of hatching eggs and conducted a detailed statistical analysis to evaluate the relationships between the studied variables. The correlation analysis revealed that eggshell mesophilic bacterial contamination (EGM) was strongly associated with yolk sac mesophilic bacterial contamination (YSM) (r = 0.76) and yolk sac contamination by Enterobacteriaceae (YSE) (r = 0.73). The principal component analysis indicated a negative association between HI performance and eggshell and yolk sac contamination. Results indicated beneficial associations between the reduction of contamination in hatching eggs and increased hatchability rates when using essential oils. The bacterial load of hatching eggs contributes to reduced productivity, reaffirming the need for proper egg sanitization, especially using essential oils. Full article

Eggs are good sources of animal proteins. Methods for creating designer eggs are attractive to scientists and producers of the eggs, in order to fulfill customers’ demands. We review and summarize the current understanding of how genetic and environmental factors influence chicken egg components, specifically, amino acid contents of yolk and albumen. Genetically diverse breeds of chicken enable us to obtain a wide variety of egg amino acids. Moreover, the use of fermented feed and different rearing systems (barn and deep litter) has impacts on egg amino acids, which may be mediated through changes in the intestinal environment, including the microbiota. To overcome the future food crisis, the combinations of breeds, feeds, and rearing systems will be important for producing designer eggs. In the future, not only long-term selected layer strains will be required, but also many kinds of indigenous chicken breeds, which have already adapted to each local environment in the world, should be investigated under the continental levels of climate environment using different fermented feed materials. To better understand the interplay between genetic and environmental factors, we will offer valuable insights for both egg producers and consumers, potentially guiding future efforts to optimize egg amino acid contents in chickens. Full article

The potential of black soldier fly larvae (BSFL) and microalgae (MA) in poultry diets has garnered increasing interest due to their high nutritional value and reduced environmental footprint. BSFL represent a sustainable alternative to conventional protein sources such as soybean meal, whereas MA contributes to improved egg quality, particularly through its enrichment with polyunsaturated fatty acids. This study assessed the effects of BSFL and MA inclusion on the growth performance and egg quality of Rhode Island Red (RIR) laying hens. Three diets were formulated: Diet A (10% BSFL), Diet B (10% BSFL + 2% MA), and Diet C (commercial control). The diets were formulated to meet the age-specific nutrient requirements of RIR hens, according to the National Research Council (NRC, 1994) guidelines. A total of 96 four-week-old chicks were randomly allocated to six pens (n = 16 per pen) and provided ad libitum access to feed and water throughout the trial. The results demonstrated that the inclusion of BSFL and MA significantly influenced the growth rate, onset of lay, and egg characteristics. Hens fed Diet B exhibited the highest average weekly body weight gain (0.034 ± 0.001 kg/week); initiated laying at 20 weeks of age, three weeks earlier than hens on Diets B and C; and produced significantly heavier eggs (51.208 ± 0.511 g). Enhanced eggshell quality and yolk pigmentation were also observed. In addition, Diet B enhanced the nutritional profile of the eggs, yielding a higher albumen protein content (76.546 ± 1.382%DM) and lower lipid concentrations (0.451 ± 0.128%DM). These findings underscore the potential of BSFL and MA as functional feed ingredients for improving poultry performance and egg quality in a sustainable production system. Full article

This review compiles information on the biochemistry of spider reproduction, from vitellogenesis to postembryonic development. Despite the diversity of spiders, biochemical studies on their reproduction remain scarce. The structures, functions, and relationships of vitellogenins and lipovitellins across different groups are compared. Information on two vitellogenin-associated proteins (30 and 47 kDa) is presented and discussed. By analyzing females at different reproductive stages—previtellogenesis, early vitellogenesis, vitellogenesis, and postvitellogenesis—as well as males, we examined lipid and fatty acid synthesis, mobilization, and accumulation in the yolk. Lipid dynamics across vitellogenic organs, such as the intestinal diverticula, hemolymph, and ovaries, were established. Structural lipids, mainly phosphatidylcholine and phosphatidylethanolamine, were the predominant yolk components, followed by triacylglycerols. The gonadosomatic and hepatosomatic indices are described for the first time in spiders, providing a new tool for studying vitellogenesis. Hemocyanin was detected in early spider eggs, suggesting a role in organogenesis, with its concentration increasing in later embryonic stages. In contrast, lipovitellin consumption was observed throughout embryonic development until juvenile emergence. The data compiled in this review provide valuable insights into the molecular interactions underlying a key process for oviparous animals. Full article

This study examined the effects of replacing corn with an aged brown rice (ABR)–wheat mixture (ABR: wheat = 85%:15%) on laying performance, egg quality, yolk fatty acid profile, economic benefits, serum biochemistry, and nutrient digestibility in laying ducks. A 12-week trial with six hundred 32-week-old Jinding ducks were randomly divided into five groups, with 10 replicates per group, and each replicate contained 12 ducks. The levels of ABR–wheat mixture in the diet were 0%, 12.5%, 25%, 37.5%, and 50%, respectively, to replace equal amounts of corn. The basic diet of the control group was corn–soybean meal (corn accounted for 50%). The experimental period was 12 weeks (from 32 to 43 weeks old). No significant differences were observed in laying performances (p > 0.05). From a numerical perspective, the duck-housed laying rate and egg mass were highest in the 37.5% group and lowest in the 50% group. Yolk color declined linearly (p < 0.05) over 12 weeks. The albumen height and Haugh unit showed a quadratic increase in weeks 4 and 8 (p < 0.05). The relative content of monounsaturated fatty acids (MUFA) increased linearly (p < 0.05), while the relative content of polyunsaturated fatty acids (PUFA), ω-3 PUFA, ω-6 PUFA, and ω-6/ω-3 PUFA decreased linearly in egg yolks (p < 0.05). The triglycerides (TG) content in serum showed a significant secondary change (p < 0.05), with the 50% group significantly lower than the other treatment groups (p < 0.05). The apparent digestibility of ether extract (EE) in the diets decreased linearly (p < 0.05). The apparent digestibility of crude protein (CP) in the diets was significantly reduced with the 50% group (p < 0.05), while there was no significant difference between the 0% and 37.5% groups (p > 0.05). Compared with the control group, economic analysis revealed net gains of 0.04–0.10 USD/duck at ≤37.5% substitution, but a loss (−0.04 USD) at 50%. In conclusion, the ABR–wheat mixture could reduce the yolk color and yolk PUFA content, but it could improve albumen quality and increase yolk MUFA content. High-level ABR–wheat mixture (50%) significantly reduced the TG content in serum and nutrient digestibility of the diet and also showed a numerical decrease in laying rate and egg mass. Our findings suggest that up to a 37.5% ABR–wheat mixture can effectively replace corn in laying duck diets without negatively affecting laying performance, while improving albumen quality and altering yolk fatty acid composition. Full article

Vaccination is widely recognized as an effective strategy for preventing various bacterial and viral diseases. In this study, protein, DNA, and egg yolk antibody (IgY) vaccines targeting the outer membrane protein VF14355 of Vibrio fluvialis (V. fluvialis) were administered to goldfish (Carassius auratus, C. auratus) subsequently challenged with V. fluvialis and Aeromonas hydrophila (A. hydrophila). The immune efficacy of the three VF14355 vaccines was evaluated through their immune activities, protective rates, anti-inflammatory and antioxidant effects, histopathology, and immunofluorescence, and the results indicated that the protective rates in the three immunized groups were significantly higher than those in the control group; furthermore, the number of kidney bacteria was significantly reduced in the immunized group compared to the control group. The ELISA results demonstrated an in vitro interaction between the bacteria and C. auratus serum. The plasma phagocytosis index and phagocytosis percentage were significantly increased in C. auratus, and their serum immune factor levels, including those of acid phosphatase (ACP), alkaline phosphatase (AKP), and lysozyme (LZM), were increased, while those of serum antioxidant factors, such as superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA), were reduced in the immunized group; notably, the expression levels of inflammatory factors were also diminished in the immunized groups. Histopathological analyses further revealed that the organ structures of the immunized group remained intact, and immunofluorescence tests indicated significant reductions in apoptosis factor p53 and DNA damage factor γH2A.X in kidney tissues. Therefore, the protein, DNA, and IgY vaccines of VF14355 demonstrate the potential to confer resistance against various bacterial infections, positioning them as promising multivalent vaccine candidates for aquaculture. Full article

Tilapia lake virus (TiLV) poses a major threat to global tilapia aquaculture and contributes to significant economic losses due to the absence of effective vaccines and treatments. Given the high mortality rates and severe pathological effects of TiLV on tilapia, alternative strategies, such as immunoglobulin-based therapies, are being considered for disease control. In this study, we developed specific immunoglobulin Y (IgY) antibodies against TiLV and evaluated their neutralization activity. Laying hens were immunized via intramuscular injections of recombinant TiLV segment 4 protein, and IgY antibodies were extracted and purified from their egg yolks using polyethylene glycol precipitation. Western blot analysis confirmed the specificity of the IgY, which demonstrated no cross-reactivity with nontarget proteins. Neutralization assays revealed a dose-dependent reduction in TiLV infectivity, which declined from 5.01 × 10⁶ TCID₅₀/mL to 5.01 × 10⁴–1.26 × 10⁵ TCID₅₀/mL, with the highest efficacy observed at a 1:2 dilution. Despite the variability in neutralization infectivity among the different hens, IgY effectively inhibited TiLV-induced cytopathic effects. Immunofluorescence assays further confirmed a significant reduction in the TiLV antigen levels in IgY-treated RHTiB cells. Our findings highlight IgY as a promising strategy for TiLV control and suggest its potential application in the prevention of emerging viruses. Full article

Two concurrent assays were conducted with the objective of evaluating the bioequivalence and determining the relative bioavailability value (RBV) of L-tryptophan biomass 60% for laying hens. In both assays, L-tryptophan 98% was used as the reference source. A total of 216 hens were used in the two assays. The first assay was performed using the direct comparison method to assess bioequivalence between the sources. In the second assay, the slope ratio method was used to determine the RBV. The variables analyzed were feed intake, tryptophan intake, egg production, egg weight, daily egg mass, feed conversion ratio, feed efficiency, yolk weight, shell weight, albumen weight, albumen height, eggshell thickness, Haugh unit, body protein weight, body lipid weight, body water weight, and body ash weight. The data from both assays were submitted to analysis of variance, and we only applied linear regression to variables from the data obtained using the slope ratio method that were significant at 0.05. The results obtained via the direct comparison method showed no significant difference (p > 0.05) between the L-tryptophan sources when using ANOVA. In the slope ratio method, the treatments only affected the egg production and daily tryptophan intake responses of the hens and linear equations were adjusted. No differences (p > 0.05) were found for egg quality. When we applied the slope ratio in adjusted equations, we found that the RBV = 100%. The results showed that the utilization of L-tryptophan biomass 60% in the diet of laying hens was bioequivalent. Full article

Insects face the constant challenge of balancing energy allocation between reproduction and immune responses, both of which are highly energy-demanding processes. Immune challenges frequently result in decreased fecundity, reduced egg viability, and delayed ovarian development. Conversely, heightened reproductive activity often suppresses immune functions. This trade-off has profound ecological and evolutionary consequences, shaping insects’ survival, adaptation, and population dynamics. The intricate interplay between reproduction and immunity in insects is regulated by the neuroendocrine and endocrine systems, which orchestrate resource distribution alongside other biological processes. Key hormones, such as juvenile hormone and ecdysteroids, serve as central regulators, influencing both immune responses and reproductive activities. Additionally, macromolecules like vitellogenin and lipophorin, primarily known for their functions as yolk protein precursors and lipid carriers, play crucial roles in pathogen recognition and transgenerational immune priming. Advancements in molecular and omics tools have unveiled the complexity of these regulatory mechanisms, providing new insights into how insects dynamically allocate resources to optimize their fitness. This delicate balance underscores critical evolutionary strategies and the integration of physiological systems across species. This review synthesizes insights from life history theory, oogenesis, and immunity, offering new perspectives on the trade-offs between reproductive output and immune investment. Full article