The effect of β-Carotene on ram semen cryopreservation has not been previously evaluated. The present research aimed to investigate the impact of different concentrations of β-Carotene in the diluent on the quality of ram semen and to analyze its antioxidant effects by the addition of tert-butyl hydroperoxide (TBHP) during cryopreservation. Semen samples were diluted with a Tris-based extender containing the β-Carotene (0, 10, 20, 30 and 40 mg/L). The motility and biokinetic characteristics, membrane and acrosome integrity, related parameters of oxidative stress, mitochondrial membrane potential (MMP) and apoptosis rate were measured after the cryo-preservation. The results indicated that the 20 mg/L β-Carotene group significantly improved total motility (TM), progressive motility (PM), average motion degree (MAD), membrane and acrosome integrity, total antioxidant capacity (T-AOC), superoxide dismutase (SOD) and catalase (CAT) compared to the other groups (P < 0.05). Additionally, this group significantly reduced reactive oxygen species (ROS) and malondialdehyde (MDA) content compared to the control group (P < 0.05). Furthermore, the TM, PM, wobble (WOB), MAD, membrane and acrosome integrity, CAT, SOD, T-AOC and MMP in the 100 μM TBHP+20 mg/ml β-Carotene group were higher than those in the 100 μM TBHP group (P < 0.05). The combined supplementation group also significantly decreased the ROS, MDA, the protein level of Cytochrome C and sperm apoptosis rates (P < 0.05). Therefore, 20 mg/L is identified as the optimal concentration for the cryopreservation of ram semen, and β-Carotene can improve the oxidative stress damage during the frozen-thawed process by enhancing the antioxidant capacity of sperm and maintaining mitochondrial function.

Reproduction in males is one of the complicated processes that is mediated by many environmental factors, as well as by diet (e.g. supplements, nutritional value). The parameters of reproduction like sperm cells, hormones and testicular histology are highly prone to oxidative stress due to influence of factors on reproductive processes. Polyphenols are naturally present in feed of animals as shrubs, trees and agro-industrial by products. The exposures of these polyphenols through diet have strong antioxidants properties. However, the benefits of these substances on reproductive health are not always certain. The use of these polyphenols in the diet of animals and in the semen extender has been reported as remarkable benefits. These beneficial effects are manifested in the form of improvement of antioxidant potential via the reduction in the production of reactive oxygen species. Polyphenols are also used for cryopreservation of various species for the genetic improvement of various species. This review gathers all the findings related to dietary consumptions of polyphenols on reproductive performance of male of various species as well as use in semen extender in fresh and frozen semen.

Moringa oleifera, a traditional Indian herb, is widely known for its capacity to induce antioxidant, anti-inflammatory and other chemoprotective effects in a broad range of biomedical models. These perspectives have led to an extensive number of studies using various moringa extracts to evaluate its capacity to protect biological systems from oxidative stress and to explore whether it could be used to slow the onset of numerous age-related conditions and diseases. Moringa extracts have also been applied to prevent damage to plants from oxidative and saline stresses, following hormetic dose–response patterns. The present paper provides the first integrated and mechanistically based assessment showing that moringa extracts commonly induce hormetic dose responses and that many, perhaps most, of the beneficial effects of moringa are due to its capacity to act as an hormetic agent.

Cadmium (Cd) is a highly harmful pollutant that poses a serious threat to human health. The liver is the primary organ for Cd accumulation, and Cd-induced hepatotoxicity has been shown to be strongly correlated with an oxidative imbalance in hepatocytes. Our previous studies in the eukaryotic model organism Saccharomyces cerevisiae revealed that not only co-treatment but also pretreatment with aqueous Moringa oleifera Lam. leaf extract (AMOLE) effectively mitigated Cd toxicity by reducing intracellular Cd accumulation and Cd-mediated oxidative stress. In this study, we therefore investigated the preventive effect of AMOLE against Cd toxicity in human HepG2 hepatocytes. The results showed that, similar to the case of the yeast model, pretreatment with AMOLE prior to Cd exposure also significantly inhibited Cd-induced oxidative stress in HepG2 cells. Untargeted LC-MS/MS-based metabolomic analysis of AMOLE revealed that its major phytochemical constituents were organic acids, particularly phenolic acids and carboxylic acids. Additionally, DPPH-HPTLC fingerprints suggested that quercetin and other flavonoids possibly contribute to the antioxidant activities of AMOLE. Based on our findings, it appears that pretreatment with AMOLE prevented Cd-induced hepatotoxicity via three possible mechanisms: i) direct elimination of free radicals by AMOLE antioxidant compounds; ii) upregulation of antioxidant defensive machinery (GPx1, and HO-1) via Nrf2 signaling cascade to improve cellular antioxidant capacity; and iii) reduction of intracellular Cd accumulation, probably by suppressing Cd uptake. These data strongly suggest the high potential of AMOLE for clinical utility in the prevention of Cd toxicity.

This study aimed at scrutinizing efficiency of incorporating L-carnitine or M. oleifera leaves extract into semen diluent on improving cryopreservation capacity and in vitro fertilization ability of buck spermatozoa. Ejaculates (n=48) were collected by an artificial vagina from six adult Damascus bucks twice weekly during the breeding season (September-October). Following initial evaluation, ejaculates of each collection session from the same bucks were pooled, diluted (1:10) with glycerolized (3 % glycerol, v/v) tris-citric acid egg yolk diluent and were split into three aliquots. The first aliquot served as control, whereas the second and third aliquots were supplemented with 4 μL/mL L-carnitine and 400 μL/mL moringa leaves extract (v/v), respectively. Thereafter, all specimens were processed for cryopreservation and were stored in liquid nitrogen (-196 °C) for 12 months before post-thaw sperm criteria were analyzed by a computer-assisted sperm analysis (CASA) system. Integrity of sperm DNA post thawing was visualized in all semen groups by fluorescence imaging, and in vitro fertilization ability of spermatozoa was also determined. Inclusion of L-carnitine or moringa leaves extract into the diluent improved (P<0.05) post-thaw sperm physical, morphofunctional and kinematic attributes, whilst maintaining (P<0.05) integrity of sperm DNA throughout the freezing and thawing cycle. Consequently, both supplemented groups yielded higher (P<0.05) in vitro fertilization rates compared to control. These results accentuate the protective roles of these antioxidants on buck sperm against consequences of cryopreservation-induced oxidative stress, hence ameliorating post-thaw sperm quality and fertilization competence. This is crucial for successful application of AI and IVF in goat selective breeding programs.

Although cryopreservation is a reliable method used in assisted reproduction to preserve genetic materials, it can stimulate the occurrence of oxidative stress, which affects sperm structure and function. This research was conducted to explore the effects of quinoa seed extracts (QSE) on ram sperm quality, oxidative biomarkers, and the gene expression of frozen-thawed ram sperm. Semen samples were diluted in extenders supplemented with 0 (QSE0), 250 (QSE1), 500 (QSE2), 750 (QSE3), and 1000 (QSE4) µg of QSE /mL, and then frozen according to the typical procedure. The findings indicate that the QSE3 and QSE4 groups provided the optimal results in terms of sperm viability and progressive motility. Sperm kinematics were considerably enhanced in the QSE3 group compared to the other groups (P<0.01). QSE (500-1000 µg/mL) significantly decreased the apoptosis-like changes (higher viable and lower apoptotic sperm) in ram sperm (P<0.001). The percentage of live sperm with intact acrosomes was significantly increased, while the percentage of detached and intact acrosomes in live and dead sperm were significantly decreased respectively by the QSE addition (P<0.001). All QSE groups had higher TAC and lower MDA and H2O2 levels than the control group (P<0.001). The expressions of SOD1, CAT, GABPB1, and GPX1 genes in sperm samples were significantly increased, while the CASP3 gene was significantly decreased in all QSE-supplemented samples. Our data suggest that QSE has beneficial effects on sperm quality of cryopreserved ram semen, which are achieved by promoting sperm antioxidant-related genes and reducing apoptosis-related gene.

The purpose of this study was to explore the effect of sodium salicylate (SS) on semen preservation and metabolic regulation in goats.

Under the condition of low temperature, SS was added to goat semen diluent to detect goat sperm motility, plasma membrane, acrosome, antioxidant capacity, mitochondrial membrane potential (MMP) and metabonomics.

The results show that at the 8th day of low-temperature storage, the sperm motility of the 20 μM SS group was 66.64%, and the integrity rates of the plasma membrane and acrosome were both above 60%, significantly higher than those of the other groups. The activities of catalase and superoxide dismutase in the sperm of the 20 μM SS group were significantly higher than those of the control group, the contents of reactive oxygen species and malondialdehyde were significantly lower than those in the control group, the MMP was significantly higher than that in the control group, and the contents of Ca2+ and total cholesterol were significantly higher than those in the control group. Through metabonomics analysis, there were significant metabolic differences between the control group and the 20 μM SS group. Twenty of the most significant metabolic markers were screened, mainly involving five metabolic pathways, of which nicotinic acid and nicotinamide metabolic pathways were the most significant.

The results indicate that SS can effectively improve the low-temperature preservation quality of goat sperm.

The aim of the present study is to evaluate the effect of Moringa oleifera leave extract (MOLE) on buffalo bull cryopreserved semen quality and fertility. Sixty ejaculates were collected from ten fertile buffalo bulls on a weekly basis for 6 weeks (n = 10 bulls & n = 60) then semen samples were pooled and divided into five groups. The semen of the control group was without additives. The semen of other groups was supplemented with MOLE at doses of 200, 400, 600 and 800 µg/ mL, respectively. One hundred thirty multiparous buffaloes were artificially inseminated with semen supplemented without or with MOLE at dose of 600 µg/ mL. Inclusion of MOLE in semen extender at dose 600 µg/ mL significantly elevated the total motility, progressive motility, membrane integrity and fertilization capacity of the post-thawed spermatozoa, as well as the total antioxidant capacity. However, it significantly decreased acrosomal defects of spermatozoa, and the concentration of malondialdehyde. This study indicated that inclusion of MOLE to semen extender improved the quality and fertility of the post-thawed buffalo bulls' semen through enhancing the activities of the antioxidant enzyme system and decreasing cryodamage of the buffalo bull spermatozoa.

To investigate the effect of Y-27632 on low-temperature metabolism of sheep sperm, different concentrations of Y-27632 were added to sheep semen at 4 °C in this experiment to detect indicators such as sperm motility, plasma membrane, acrosome, antioxidant performance, mitochondrial membrane potential (MMP), and metabolomics. The results showed that the addition of 20 µM Y-27632 significantly increased sperm motility, plasma membrane integrity rate, acrosome integrity rate, antioxidant capacity, MMP level, significantly increased sperm adenosine triphosphate (ATP) and total cholesterol content, and significantly reduced sperm Ca2+ content. In metabolomics analysis, compared with the control group, the 20 µM Y-27632 group screened 20 differential metabolites, mainly involved in five metabolic pathways, with the most significant difference in Histidine metabolism (P = 0.001). The results confirmed that Y-27632 significantly improved the quality of sheep sperm preservation under low-temperature conditions.

This study was planned to detect the adverse pathological consequences of aflatoxin B1 in White Leghorn (WLH) layer breeder males. Eight-week-old male layer cockerels were separated into six experimental categories: A group was kept as negative control, offered with normal feed only; group B was fed with 400 ppb amount of aflatoxin, while groups F and D fed with normal feed and supplemented with vitamin E 100 ppm and 1% Moringa oleifera, respectively, whereas groups E and C were fed with 400 ppb aflatoxin containing feed and ameliorated with vitamin E 100 ppm and 1% Moringa oleifera, respectively. This study was continued for 2 months and immunologic disorders and reproductive parameters were observed during the trial. To find out immunological status lymphoproliferative response to phytohemagglutinin-P (PHA-P), antibody titers against sheep red blood cells (SRBCs) and carbon clear assay were performed by collecting samples from five birds from each group. The whole data was measured by ANOVA test, and group means were compared by DMR test by using M-Stat C software. Regarding the reproductive status, spermatogenesis, blood testosterone level, testes weight, testes histology, sperm motility, and morphology were negatively affected by aflatoxins, but these deviations positively ameliorated by vitamin E and Moringa. Vitamin E and Moringa found advantageous in boosting the immune status of affected bird. All the immunological parameters including antibody titers against sheed red blood cells, lymphoproliferative response to avian tuberculin and phagocytic potential of macrophages were suppressed by AFB1 however in control, Moringa and vitamin E groups these immunological responses were significantly higher.

Drastic climatic changes threaten animal productivity and prolificacy, whose adaptability is governed by its pheno- and genotypic traits. This study was aimed at investigating the effect of coat color on the adaptability of goat bucks under heat stress conditions from the perspectives of testicular blood flow (TBF) and biometry, reproductive hormones, and semen quality. Twenty bucks (Capra hircus) bearing different coat colors were selected from a large flock and divided into four equal groups (n = 5 each) as follows: black coat (BC; 100% black), brown coat (BrC; 100% dark brown), white coat (WC; 100% white), white-black coat (WBC; 50-60% white). Bucks were examined for TBF [Doppler ultrasonography and serum nitric oxide (NO)], testosterone (T) and luteinizing hormone (LH), seminal plasma oxidative biomarkers [catalase (CAT), total antioxidant capacity (TAC), and malondialdehyde (MDA)], and sperm traits percentages [progressive motility (PM), viability (SV), normal morphology (NM), and sperm concentration (SC) once a week for seven consecutive weeks (W1-W7) in the summer season (temperature humidity index = 88.4-92.2). Specifically, at W3-W7, darker bucks (BC and BrC) testicular volume, testicular colored area, T, NO, CAT, TAC, PM, SV, NM, and SC (W7 only) differed significantly (P < 0.05) by decrease than the lighter ones (WC and WBC). Both Doppler indices and serum MDA concentrations were elevated (P < 0.05) at W3-W7 in the BC and BrC bucks compared to WC and WBC groups. In conclusion, bucks with lighter coats were more resistant to the negative effects of HS on TBF, seminal oxidative biomarkers, and semen quality.

Abamectin (ABM), a naturally fermented product of Streptomyces avermitilis, is applied to pest control in livestock and agriculture fields. The aim of the current study is to evaluate the protective effects of Moringa oleifera leaf ethanolic extract (MOE) on biochemical changes including oxidative stress indices, immune response marker, lipid profiles as well as mRNA expression of immune related genes, and abamectin (ABM, 5% EC) residue levels in Nile tilapia (Oreochromis niloticus) exposed to a sub-lethal concentration (0.5 µg/l) for 28 days. Disturbance in liver and kidney biomarkers was markedly increased in ABM-exposed fish compared to the control group. Malondialdehyde levels in the liver and brain tissues, as well as the activities of glutathione-s-transferase, superoxide dismutase, and glutathione peroxides, all increased significantly in ABM group. Additionally, ABM exposure increased the levels of interleukin 10 beta and growth factor gene expression. On the other hand, fish exposed to ABM had significantly lower serum alkaline phosphatase, creatinine, high-density lipoprotein, glutathione peroxides in brain, glutathione in liver and brain tissues, lysozyme activity, nitric oxide, immunoglobulin M, tumor necrosis factor, and interleukin 1 beta as compared to the control group. The recorded detrimental effects of ABM on tilapia have been overcome by the addition of MOE to the diet (1%) and ameliorating hepato-renal damage and enhancing antioxidant activity, innate immune responses, and upregulating the anti-inflammatory gene expression. Therefore, it could be concluded that MOE dietary supplementation at 1% could be used to counteract the oxidative stress, immune response disruption induced by abamectin exposure in Oreochromis niloticus, and reduce its accumulation in fish tissues.

This study aimed to determine the effects of melatonin administration on testicular vascular perfusion in relation to steroid hormones and semen characteristics in dogs. The study included 12 normospermic German shepherd dogs (weighed 35 ± 0.5 kg and aged 4 ± 0.5 years). Males received a single melatonin administration (melatonin dimethyl sulfoxide + corn oil via subcutaneous route; MEL; n = 6), while the rest of the animals served as controls (dimethyl sulfoxide + corn oil; Control; n = 6). Males were subjected to routine examination on days -15, 0, 15, 30, 45, and 60. All examined dogs were subjected to Doppler screening, semen collection, and blood sampling. The MEL group showed a significant (P < 0.05) elevation in semen volume, concentration, percentage of sperm motility, and total sperm × 106 / ejaculate compared to other control males. Doppler indices as resistance (RI) and pulsatility (PI) indices declined (P < 0.05) from D 30 (1.02 ± 0.01) until day 60 (0.87 ± 0.02) of treatment. In MEL males, the peak systolic point of velocity (PSV; cm/sec) of the testicular artery elevated (P < 0.05) on day 60 (20.15 ± 0.99) compared to its value on day 0 (17.39 ± 1.84). On D 60, the levels of testosterone (T), estradiol 17-ß (E2), and nitric oxide (NO) elevated (P < 0.05). A negative correlation was detected between testicular volume, scrotal circumference (SC), T levels, Doppler indices, and velocities. In conclusion, single melatonin administration could improve testicular vascularization via increasing Doppler velocities and intratesticular colored areas. In addition, it could improve semen picture and steroids (T and E2) and nitric oxide.