Melatonin, an antioxidant hormone secreted by the pineal gland, has been recognized as a regulator for numerous biological events. The deleterious effects of juglone, a polyphenolic extract of walnut trees, on embryo development has been previously reported. In the current study, we aimed to display the impact of melatonin administrated during in vitro oocyte maturation (IVM) on juglone-treated oocytes. Thus, in vitro matured oocytes were collected after 24 h post incubation with juglone in the presence or absence of melatonin. Reactive oxygen species (ROS), glutathione (GSH) content, mitochondrial distribution, and the relative abundance of mRNA transcription levels were assessed in oocytes, in addition, oocytes were in vitro fertilized to check the competency levels of oocytes to generate embryos. We found that administration of melatonin during the maturation of oocytes under juglone stress significantly improved the cleavage rate, 8-16 cell-stage embryos and day-8 blastocysts when compared to the sole juglone treatment. In addition, the fluorescence intensity of ROS increased, whereas the GSH decreased in juglone-treated oocytes compared to melatonin–juglone co-treated and untreated ones. Additionally, a significant increase in the mitochondrial aberrant pattern, the pattern that was normalized following melatonin supplementation, was observed following juglone administration. The mRNA analysis using RT-qPCR revealed a significant upregulation of autophagy and oxidative-stress-specific markers in the juglone-treated group compared to the co-treatment and control. In conclusion, the study reveals, for the first time, a protective effect of melatonin against the oxidative stress initiated following juglone treatment during the in vitro maturation of oocytes.

Understanding the metabolic profile within the follicular microenvironment is crucial for optimizing reproductive efficiency in camels. In this study, we examined the metabolomic profile of camel follicular fluid (FF) during the breeding (n = 10) and non-breeding seasons (n = 10). Gas chromatography-mass spectrometry (GC-MS) was utilized to describe the metabolites present in follicular fluid samples. The results found considerable differences in the metabolomics profiles between the breeding and non-breeding seasons. Hexadecenoic acid, galactose and glucose levels were significantly (P < 0.05) higher in camel FF during the breeding season, while 9-octadecenamide, oleonitrile, glycine, octadecanamide, cholesterol, and propanoic acid were higher (P < 0.05) in FF during the non-breeding season. Multivariante analyses pointed to those 9 metabolites, and univariate analysis showed hexadecenoic acid, galactose, glucose, and oleanitril were the most significant ones in camel follicular fluid collected during both breeding and non-breeding seasons. The univariate and multivariate analyses showed an increase in the levels of hexadecanoic acid, galactose, glucose, and a depletion in the level of oleanitrile in the breeding season compared to the non-breeding season. The ROC curve and statistical analysis showed that hexadecanoic acid, galactose, and oleanitril with AUC = 1 were promising to be seasonal biomarkers of fertility in female camels. In conclusion, the metabolomic analysis of camel FF reveals distinct changes in metabolite levels between breeding and non-breeding seasons, reflecting adaptive metabolic responses to support reproductive processes. These results offer valuable insights into the reproductive physiology of camels and offer practical implications for potential biomarkers and assessing the reproductive status in camels, which can be utilized in reproductive management and conservation efforts in these valuable animal species.

Background: Aloe vera gel is rich in polysaccharides (acemannan), phenolic compounds, vitamins, and minerals, while Pomegranate peel ( Punica granatum L.) is a valuable source of punicalagin, polyphenols, tannins, flavonoids, dietary fiber, vitamins, and minerals. This study examined the impact of these bioactive-rich supplements on growth performance, metabolic activity, digestive enzyme function, antioxidant status, levels of nucleic acids and proteins, as well as gastrointestinal histomorphometry in growing rabbits. Materials and methods: Thirty male New Zealand White rabbits (56 ± 3 days) at the start of the trial, corresponding to the early post-weaning/growing phase in New Zealand White rabbits. Rabbits were randomly divided into three groups ( n = 10) and treated for 14 weeks. The control group (C) received a basal diet; the pomegranate group (P) received the basal diet supplemented with 4.5% pomegranate peel; and the Aloe vera group (A) received the basal diet with drinking water containing Aloe vera gel (500 mg/L). Results: Both supplemented groups exhibited significantly higher body weight and improved metabolic indices, including elevated blood glucose, total protein and lipid profile, compared with the control group ( p < 0.05). Activities of amylase, lipase, and protease increased significantly, with stronger stimulation observed in the pomegranate group. Antioxidant assays revealed higher total antioxidant capacity (TAC) and catalase (CAT) activity, accompanied by reduced malondialdehyde (MDA) levels in both supplemented groups ( p < 0.05). DNA and total protein concentrations were also elevated, particularly in the pomegranate group. Histomorphometric analysis of the duodenum showed significant improvements in villus height, crypt depth, and glandular area ( p < 0.05). Aloe vera supplementation exerted greater effects on villus and crypt architecture, whereas pomegranate peel predominantly enhanced glandular development. Molecular docking simulations revealed that acemannan and punicalagin possess high binding affinities for pro-apoptotic and antioxidant targets. Specifically, acemannan exhibited markedly lower binding energies than punicalagin for both BAX (−10.627 vs. –7.540 kcal/mol) and SOD (−10.544 vs. –7.663 kcal/mol). These results suggest that acemannan may exert superior bioactivity by effectively modulating BAX-mediated apoptosis and augmenting SOD-driven antioxidant defense through stable protein-ligand complexation. Discussion: In conclusion, dietary supplementation with pomegranate peel or Aloe vera significantly improved growth performance, optimized metabolic activity, and enhanced intestinal morphology in growing rabbits. Each supplement provided unique physiological benefits, supported by molecular docking evidence linking their bioactive compounds to antioxidant and cytoprotective mechanisms.