Search Results (49)

The physiological functions of mammalian sperm, such as motility, hyperactivation, and capacitation, require substantial energy. This study investigates the effects of two classic cryopreservation extenders—TCG (tris-citrate-glucose) and LEY (lactose-egg yolk)—on the energy metabolism of frozen–thawed boar semen. By comparing the quality indicators, key metabolite levels, and the activities of critical enzymes involved in glycolysis and the tricarboxylic acid cycle, we aim to understand how these different semen extenders influence the spermatozoa vitality of frozen–thawed boar semen. Following thawing, the LEY-cryopreserved sperm demonstrated significantly elevated motility parameters (viability, VCL, VSL, and VAP) and enhanced plasma membrane and acrosomal integrity compared with the TCG group (p < 0.05), though both cryopreserved groups exhibited significantly reduced performance relative to fresh semen controls. Cryopreservation markedly reduced intracellular adenosine triphosphate (ATP), pyruvate, and acetyl coenzyme A (A-CoA) levels (fresh > LEY > TCG; p < 0.05). The LEY-preserved spermatozoa retained higher activities of glycolysis-related enzymes (phosphofructokinase, PFK; pyruvate kinase, PK) compared with the TCG group, which, in turn, showed elevated lactate dehydrogenase (LDH) activity. Critically, TCG-suppressed pyruvate dehydrogenase (PDH) activity (p < 0.05) coincided with diminished A-CoA, indicating impaired mitochondrial oxidative phosphorylation. These results demonstrate LEY’s superior preservation of motility and membrane stability but highlight cryodamage-induced energy metabolism dysregulation, particularly TCG’s disruption of the glycolysis–TCA cycle coordination essential for spermatozoa function. In conclusion, the choice of semen extender has a significant impact on the energy metabolism and overall quality of frozen–thawed semen, highlighting the importance of optimizing cryopreservation protocols for improved spermatozoa viability and functionality. Full article

The study aimed to evaluate the effectiveness of the cryopreservation protocols for Pseudoplatystoma magdaleniatum semen using dimethyl sulfoxide (DMSO) or methanol (MET) as permeable cryoprotectants at two concentrations (5% and 10%) combined with 12% egg yolk (Y12%) or 5% skimmed milk powder (SMP5%) and glucose (6%), resulting in eight treatments. A semen pool (n = 8) was diluted in a 1:4 ratio, packed in 2.5 mL straws, and frozen in nitrogen vapors. It was thawed at 35 °C for 90 s. Sperm kinetics and motility duration of fresh, prefrozen, and thawed semen were analyzed using a CASA system. The osmolarity of seminal plasma and cryosolutions was estimated. Fertilization (F) and embryo viability (E) rates of thawed semen were evaluated. The osmolarity of seminal plasma was 251.1 ± 3.3 mOsmol/kg and, in the cryosolutions, ranged between 1248.3 ± 19.9 mOsmol/kg (DMSO5% + Y12%) and 3488.2 ± 1.5 mOsmol/kg (MET10% + Y12%). After thawing, total motility ranged from 38.2% to 60.5%, representing a significant reduction compared to fresh semen (95.4 ± 2.1%) (p < 0.05). The best fertilization and embryo viability rates of thawed semen were obtained with DMSO5% + SMP 5% (F = 20.7%, E = 11.7%) and MET10% + SMP5% (F = 20.1%, E = 11.5%) (p < 0.05). A cryopreservation protocol for P. magdaleniatum semen with 5%DMSO or 10%MET combined with SMP5% is possible, but further study is necessary to optimize its fertilizing capacity. 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

Egg yolk, commonly employed as a cryoprotectant in semen cryopreservation, contains large particle matter that can diminish semen quality post thaw and complicate its quality assessment. For this reason, we designed a centrifugal treatment of chicken egg yolk to evaluate its effect on the cryopreservation of porcine semen. The control group (CG) was prepared with a dilution of chicken egg yolk by conventional mixing treatment, and the experimental group (EG) used a dilution of centrifugally treated chicken egg yolk for the ultra-low-temperature cryopreservation of porcine semen. The freezing process was carried out by conventional freezing methods. The spermatozoa were subsequently assessed for various parameters, including motility, acrosome integrity rate, plasma membrane integrity rate, antioxidant indexes, apoptosis rate, and the expression of apoptosis-related genes. The results showed that, post freeze–thawing, the motility, viability, VSL, and VCL of the spermatozoa in the EG were significantly higher than those observed in the CG (p < 0.05). Additionally, the acrosome integrity and plasma membrane integrity of the spermatozoa in the EG were significantly enhanced compared to the CG (p < 0.05). Furthermore, the EG exhibited significantly lower MDA content and sperm apoptosis rate (p < 0.05), while demonstrating significantly higher T-AOC and CAT levels (p < 0.05) relative to the CG. In comparison to the CG, the EG exhibited a significant reduction in the gene expression of TNF-a and Bax in the spermatozoa (p < 0.05), whereas the expression levels of CAT and Bcl-2 were significantly elevated (p < 0.05). In conclusion, the dilution solution formulated through the centrifugal processing of chicken egg yolk demonstrated efficacy in enhancing the quality of porcine spermatozoa following cryopreservation and subsequent thawing. Full article

Carotenoids from different sources have different structures and functions, and their dietary components benefit the health of various organisms. The effects of halophilic Archaea-derived C50 carotenoids on poultry egg quality and gut microbiota remain largely unexplored. In this study, we isolated a carotenoid-secreting strain of Halalkalicoccus paucihalophilus, TRM89021, from the Pamir Plateau. We characterized the carotenoid pigments produced by this strain; the major components were bacterioruberin and its derivatives. The effects of these carotenoids on the egg quality and cecal microbiota composition of hens were investigated. Compared to the basal diet group (BDG), supplementation with carotenoids in the carotenoids-supplemented diet group (CDG) resulted in significantly lower a* and b* scores at week 5 and lower b* scores and Haugh units at week 2, while egg strength and weight were higher. CDG also showed increased yolk antioxidant capacity, higher glutathione peroxidase levels, and significantly lower catalase levels (p < 0.05). Plasma analysis revealed elevated total bilirubin and aspartate aminotransferase levels, along with reduced inorganic phosphorus levels in the CDG (p < 0.05). No significant differences in cecal microbiota diversity were observed between the groups at any taxonomic level. This result suggests that halophilic archaea-derived carotenoids have the potential to be used as natural feed supplements to improve egg quality. Our study provides a theoretical basis for applying archaea-derived carotenoids in poultry diets. Full article

Producing translucent eggs has been found to reduce the quality and safety of the eggs, as well as the demand from consumers. However, the intestinal function and the molecular mechanism for the production of translucent eggs remain uncertain. A total of 120 eggs from 276-day-old Jining Bairi were divided into two groups based on eggshell translucence: the translucent egg group (group T) and the normal group (group C). Group T exhibited thicker eggshells and a lower egg yolk color. Subsequently, we divided the chickens into translucent and normal groups based on their egg quality. We then assessed the plasma biochemical index, intestinal morphology and structure, enzyme activity, and antioxidant capacity of the hens producing translucent eggs compared to those producing normal eggs. The results showed that the ratio of duodenal villus length to crypt depth, succinate dehydrogenase (SDH) activity, chymotrypsin, total ATPase (T-ATPase), alkaline phosphatase (AKP), and glutathione peroxidase (GSH-Px) activities were decreased in the hens produced translucent eggs (p < 0.05), but malondialdehyde (MDA) content was increased (p < 0.05); jejunal lipase activity, Na⁺K⁺-ATPase activity, total antioxidant capacity (T-AOC), and GSH-Px activities were decreased (p < 0.05) in group T; ileal amylase and Ca²⁺Mg²⁺-ATPase activities were also decreased (p < 0.05) in group T. In addition, we identified a total of 471 differentially expressed genes (DEGs) in duodenal tissue, with 327 up-regulated genes and 144 down-regulated genes (|log₂FC| ≥ 1 and p < 0.05). Enrichment analysis showed that the up-regulated genes, such as GSTT1, GSTO2, and GSTA3, were mostly enriched in metabolism of xenobiotics by cytochrome P450, drug metabolism-cytochrome P450, and oxidative phosphorylation pathways. The results of our study indicate that plasma lipid metabolism disorder, decreased intestinal antioxidant capacity, and altered intestinal metabolism capabilities may influence the formation of translucent eggs. Full article

This study aimed to investigate the effects of dietary riboflavin levels on the reproductive performance of pigeon breeders and the growth performance and carcass traits of offspring squabs to estimate the riboflavin requirement of pigeon breeders. The natural riboflavin content in the basic diet of corn–peas–soybean–wheat–sorghum–corn gluten is 1.20 mg/kg. Different doses of riboflavin (0, 2.5, 5, 10, and 15 mg/kg) were supplemented with the basal diet to produce five dietary treatments with total riboflavin levels of 1.20, 3.70, 6.20, 10.20, and 16.20 mg/kg. A total of 120 pairs of White King pigeons, aged 60 wks, were randomly allocated into five treatment groups, each consisting of 24 pairs. Each pair was individually raised for 8 wks. After the experiment, an assessment was conducted to evaluate the reproductive performance of the pigeon breeders, as well as the growth and carcass traits of offspring squabs at 28 days of age. The results showed that the dietary riboflavin levels had no significant effect on body weight, feed intake, egg weight, egg production, and egg fertility (p > 0.05). However, pigeons fed a diet without riboflavin had the lowest egg hatchability, egg yolk color, carcass trait, and riboflavin status, while exhibiting higher liver weight and liver index (p < 0.05). Moreover, the indices above showed increased or decreased linearly as the level of riboflavin was increased in the diet. Based on the broken-line regression model, pigeon breeders were determined to require a dietary riboflavin content of 11.4, 13.6, 13.4, 6.60, 4.28, 4.47, 4.67, 6.69, and 6.82 mg/kg to optimize hatchability, eviscerated weight, half-eviscerated weight, breast muscle weight, breast muscle percentage, liver weight, liver index, egg yolk riboflavin, and squab plasma riboflavin, respectively. In conclusion, the optimal supplemental dosage of riboflavin in the diets of pigeon breeders is 13.6 mg/kg. Full article

Red light (RL) has been observed to enhance egg production in pigeons, yet the underlying histological characteristics and molecular mechanisms remain less understood. This study included fifty-four pigeons to assess follicular histology, reproductive hormones, and ovarian transcriptomics on the third day of the laying interval under RL and white light (WL). The results showed that the granulosa cell layer was significantly thicker under RL (p < 0.05), whereas the theca cell and connective tissue layers showed no significant differences (p > 0.05). Higher plasma estradiol (E₂) levels were recorded in the RL group (p < 0.05), as well as follicle stimulating hormone (FSH), although progesterone (P₄) levels were higher under WL (p < 0.05). Moreover, P₄ concentrations in follicle yolk significantly decreased under RL (p < 0.01), with higher FSH and E₂ levels in F1 yolk and similar increases in SF1 yolk (p < 0.01). Transcriptomic analysis revealed 4991 differentially expressed genes in the pigeon ovary. The protein–protein interaction network highlighted genes like HSD11B1, VEGFD, WNT6, SMAD6, and LGR5 as potential contributors to hierarchical follicle selection under RL. This research provides new insights into the molecular basis by which RL may promote hierarchical follicle selection and improve egg production in pigeons. Full article

Antioxidants protect cellular function and structure by neutralizing the oxidative stress caused by increased reactive oxygen species (ROS) during sperm freezing. Studies on cryopreservation using various antioxidants have demonstrated encouraging results. Many studies have used antioxidants to increase the efficiency of sperm freezing and to improve the success rate of artificial insemination and pregnancy. Manganese (III) tetrakis (4-benzoic acid) porphyrin chloride (MnTBAP) is a newly synthesized antioxidant with positive effects on sperm morphology and capacitation in humans, rams, and stallions. In this study, porcine semen was treated with 0, 50, 100, and 150 μM of MnTBAP based on a Tris–egg-yolk extender and frozen to determine whether MnTBAP can assist the status of sperm during cryopreservation. First, motility was assessed using the computer-assisted sperm analysis (CASA) system, with the 100 μM treatment group showing the highest motile rate (66.8%) compared with that of the other groups (control, 51.1%; 50 μM and 150 μM, 59.6%); therefore, the remaining analyses were conducted comparing the two groups (control vs. 100 μM group; p < 0.01). Second, fluorescence staining was applied to examine the control and 100 μM groups using fluorescence microscopy. The viability (41.7% vs. 62.4%) and the acrosome integrity (77.9% vs. 86.4%) differed significantly (p < 0.05). In addition, the mitochondrial membrane potential (MMP) was 46.5% vs. 51.9%; the fragmentation rate, estimated using the Sperm-sus-Halomax kit, was 63.4% vs. 57.4%; and the detected caspase activity was 30.1% vs. 22.9%. These tended to be higher in the treated group but did not differ significantly. Third, measurements using FACSLyric revealed that the 100 μM treatment group exhibited a state of elevated normal lipid arrangement within the plasma membrane and diminished levels of apoptosis and ROS (p < 0.01). We assessed the expression of genes relevant to antioxidant effectiveness using real-time RT-qPCR. Our findings indicated significant alterations in the expression levels of various mRNA species, with the exception of NOX5 (p < 0.05). Finally, the straws were dissolved and used to treat matured denuded oocytes to investigate the effect on fertilization and embryo development in vitro. The cleavage rate was (77.6% vs. 84.1%), and the blastocyst rate was 9.7% vs. 11.4% (p < 0.05). In conclusion, these results suggest that MnTBAP positively affected sperm freeze–thawing, improving the fertilization capacity, and leading to increased embryo development. Full article

The aim of this experiment was to investigate the effects of dietary Phytosterol Ester (PSE) supplementation on egg characteristics, eggshell ultrastructure, antioxidant capacity, liver function, hepatic metabolites, and its mechanism of action in Hy-Line Brown laying hens during peak laying period. A total of 256 healthy Hy-Line Brown laying hens were randomly allocated into four groups. The hens in the control group were fed a basal diet, while those in the experimental groups were fed a basal diet further supplemented with 10, 20, and 40 mg/kg PSE, respectively. It was found that the addition of 20 mg/kg and 40 mg/kg PSE to the diets increased egg weight, but decreased egg breaking strength (p < 0.05). The addition of PSEs to the diets increased albumen height and Haugh unit in all experimental groups (p < 0.05). Electron microscopic observation revealed that the mammillary thickness increased significantly at doses of 20 and 40 mg/kg, but the total thickness decreased, and the effective thickness also thinned (p < 0.05). The mammillary width narrowed in all experimental groups (p < 0.001). Dietary supplementation with 40 mg/kg PSE significantly increased egg yolk Phenylalanine, Leucine, and Isoleucine levels (p < 0.05). In untargeted liver metabolomic analyses, L-Phenylalanine increased significantly in all experimental groups. Leucyl-Lysine, Glutamyl-Leucyl-Arginine, and L-Tryptophan increased significantly at doses of 10 and 20 mg/kg (p < 0.05), and L-Tyrosine increased significantly at doses of 10 and 40 mg/kg (p = 0.033). Aspartyl-Isoleucine also increased significantly at a dose of 10 mg/kg (p = 0.044). The concentration of total protein in the liver was significantly higher at doses of 20 and 40 mg/kg than that of the control group, and the concentrations of total cholesterol and low-density lipoprotein cholesterol were significantly reduced (p < 0.05). The concentration of triglyceride and alkaline phosphatase were significantly reduced in all experimental groups (p < 0.05). Steatosis and hemorrhage in the liver were also improved by observing the H&E-stained sections of the liver. Concerning the antioxidant capacity in the liver, malondialdehyde concentration was significantly reduced (p < 0.05) at a dose of 40 mg/kg. In the ovary, malondialdehyde and nitric oxide concentrations were significantly reduced (p < 0.001). In all the experimental groups, plasma nitric oxide concentration was significantly decreased while superoxide dismutase was significantly increased, and total antioxidant capacity concentration was significantly increased (p < 0.05) in the 10 mg/kg and 40 mg/kg doses. Metabolomics analyses revealed that PSEs play a role in promoting protein synthesis by promoting Aminoacyl-tRNA biosynthesis and amino acid metabolism, among other pathways. This study showed that the dietary addition of PSEs improved egg characteristics, antioxidant capacity, liver function, and symptoms of fatty liver hemorrhagic syndrome in Hy-Line Brown laying hens at peak laying stage. The changes in liver metabolism suggest that the mechanism of action may be related to pathways such as Aminoacyl-tRNA biosynthesis and amino acid metabolism. In conclusion, the present study demonstrated that PSEs are safe and effective dietary additives as an alternative to antibiotics. Full article

The dilution method and ratio were tested to assess their effects on the Hu ram semen after cryopreservation. Experiment I aimed to explore the effect of various dilution ratios (1:1, 1:2, 1:3, 1:4) of diluent I (Tris-based and egg yolk) under the condition of 1:1 dilution of diluent II (diluent I and glycerol) on the Hu ram semen preserved in liquid nitrogen regarding spermatozoa motility and kinetic parameters. Experiment II aimed to investigate the effect of various dilution ratios (1:1, 1:2, 1:3, 1:4) of diluent I under the condition of 1:2 dilution of diluent II to the Hu ram semen for cryopreservation on spermatozoa motility and kinetic parameters. The purpose of experiment III is to assess the effect of various dilution methods and ratios on the cryopreservation of Hu ram semen by detecting spermatozoa motility, kinetic parameters, plasma membrane integrity, acrosome integrity and reactive oxygen species (ROS) level. Experiment III includes four groups: one-step dilution method and two-step dilution method. The two-step dilution method includes two groups: 1:2, 1:1 and 1:3, 1:2, and the one-step dilution method includes two groups: 1:5 and 1:11. The results indicated that the post-thawed spermatozoa total motility (TM), progressive motility (PM) and average motion degree (MAD) were highest in the 1:2 group and significantly higher (p < 0.05) than those in the 1:1 and 1:4 groups under the condition of 1:1 dilution of diluent II. The post-thawed spermatozoa TM and PM of the 1:3 group were significantly higher (p < 0.05) than those of the other groups under the condition of 1:2 dilution of diluent II. The post-thawed spermatozoa TM, PM, plasma membrane integrity and acrosome integrity of the two-step group (1:3, 1:2) were the highest and significantly higher (p < 0.05) than those in the other groups. Additionally, the post-thawed spermatozoa ROS level of the two-step group (1:3, 1:2) was significantly lower (p < 0.05) than that in the one-step groups (1:5 and 1:11). Therefore, a two-step dilution (1:3, 1:2) was found to be the most suitable method and ratio for diluting the Hu ram semen after cryopreservation. Full article

Lithium (Li) is present in human nutrition based on food intake, and several studies recommend it for treating mood disorders, even if the biological proprieties and biochemical mechanisms represent the basis for its use as an essential element. The Li content was evaluated using the inductively coupled plasma mass spectrometry technique (ICP-MS) in 1071 food and beverage samples from the Romanian market. The results show that Li had a decreasing mean concentration in the food samples as follows: vegetables leafy > bulbous > fructose > leguminous > egg whites > root vegetables > milk products > egg yolks > meats. Approximately a quarter of all data from each dataset category was extreme values (range between the third quartile and maximum value), with only 10% below the detection limit. Mean Li concentration indicated higher values in red wine, white wines, beers, and fruit juice and lower in ciders and bottled waters. A particular interest was addressed to plants for teas and coffee seeds, which showed narrow amounts of Li. For both food and beverages, two similar matrices, including egg whites and yolks and white and red wines, were found to have significant differences, which explains the high variability of Li uptake in various matrices. For 99.65% of the analyzed samples, the estimated daily intake of Li was below the provisional subchronic and chronic reference dose (2 µg/kg_bw/day) for adverse effects in several organs and systems. Even so, a risk occurs in consuming bulbous vegetables (Li > 13.47 mg/kg) and fructose solano vegetables (Li > 11.33 mg/kg). The present study’s findings indicate that ingesting most of the analyzed beverages and food samples could be considered safe, even if future studies regarding Li content, nutritional aspects, and human cohort diseases must be conducted. Full article

Yolkin is a polypeptide complex isolated from hen egg yolk that exhibits immunomodulating properties. The aim of the present study was to determine whether in-ovo-delivered yolkin affects leukocyte populations and cytokine levels in broiler chickens. The experiment was carried out on eggs from Ross 308 broiler breeder birds. Yolkin was administered in ovo on the 18th day of incubation, once, at the following three doses: 1, 10, or 100 µg/egg. The immunological parameters were assessed in 1-, 7-, 14-, 21-, 28-, 35-, and 42-day-old birds kept under farming conditions and routinely vaccinated. The leukocyte populations were determined in the thymus, spleen, and blood. The cytokine (IL-1β, IL-2, IL-6, and IL-10) levels were determined in the plasma of the broiler chickens. Each experimental group included eight birds. The most pronounced effect of yolkin was an increase in the population of T cells, both CD4⁺ and CD8⁺, mainly in the blood. This effect on the lymphocyte subsets may be valuable regarding chicken immune responses, mainly against T-dependent antigens, during infection or after vaccination. Full article

This study aimed to investigate the effect of enriching ice cream with high biological value animal protein sources from by-products on its techno-functional properties. Ice creams were prepared with 10 g (100 g)⁻¹ amount of different enrichments: whole egg, egg yolk, egg white, animal blood plasma, whole blood, and haemoglobin. The rheological properties, frozen texture, colour, dry matter content, and pH of the ice cream samples were analysed. The results indicate that these ‘natural food additives’ can influence the quality properties of ice cream, highlighting the potential for developing novel ice cream products with enhanced nutritional value and decreasing food wastes. The study reveals that enrichments significantly affect the rheological attributes of the ice cream mix, altering yield stress and consistency index without changing the overall rheological behaviour. The frozen texture varies among enrichments, with egg white resulting in the softest texture, while blood-based enrichments and egg yolk contribute to a harder texture. Colour analysis indicates changes in redness/greenness, yellowness/blueness and lightness due to different pigments and foaming effects of enrichment materials. Furthermore, enrichments influence dry matter content and pH, with egg yolk increasing fat content and haemoglobin boosting protein content. Sensory analysis suggests that certain enrichments improve taste sensation and colour preference, making them more acceptable to consumers despite the by-product ingredients. Full article

Egg is a food product of high nutritional quality, extensively consumed worldwide. The objectives of this study were the determination of the elemental profile in eggs (egg white, yolk, and eggshell), the estimation of the non-carcinogenic health risk associated with the presence of heavy metals in investigated egg samples, and the development of statistical models to identify the best predictors for the differentiation of egg components. The assessments were carried out in a total set of 210 samples, comprising home-produced and commercial eggs, using inductively coupled plasma mass spectrometry. The results suggested measurable differences amongst hen eggs coming from different husbandry systems. The statistical models employed in this study identified several elemental markers that can be used for discriminating between market and local producer samples. The non-carcinogenic risk related to the consumption of the analyzed egg samples was generally in the safe range for the consumers, below the maximum permitted levels set by Romanian and European legislation. Food contamination is a public health problem worldwide, and the risk associated with exposure to trace metals from food products has aroused widespread concern in human health, so assessing the heavy metal content in food products is mandatory to evaluate the health risk. Full article