Search

The effect of dietary calcium (Ca) and magnesium (Mg) supplementation on serum biochemical parameters, steroid hormones, gene expression, and the sex ratio was investigated in female New Zealand white rabbits. A total of 25 rabbits were allocated into five treatment groups: The control group was fed with regular pellet feed, whereas, treatment groups were supplemented with Ca and Mg: T1 (0.40% and 0.01%), T2 (0.60% and 0.02%), T3 (0.80% and 0.03%) and T4 (1.00% and 0.04%), respectively. The rabbits were subjected to three breeding cycles. The T3 group skewed towards females (65.33%) from all three breeding. There was elevated Ca concentration in T3 (15.26 ± 0.77 mg dL-1) and T4 (15.61 ± 0.82 mg dL-1) groups compared to the control. The concentration of estradiol was significantly high in T3 and T4 groups at 0.5 days post-coitus (dpc) and T2, T3 and T4 groups at 21dpc. Testosterone was significantly high in T4 group at 0.50 dpc and T2 and T4 group at 21dpc. The expression of 13 genes was studied in the oviduct. Genes such as OVGP1, CCT4, ANXA2 and TLR4 were upregulated and positively correlated with the female sex ratio. The molecular functions and pathways of up-regulated genes were suggestive of their role in fertilization such as sperm selection, sperm storage, immune regulation, implantation and early embryonic development. The variations in the serum electrolytes, steroid hormones and gene expression might have an impact on the skewing process. [ABSTRACT FROM AUTHOR]

Abstract The reproductive performance during the transition from prepubertal to pubertal stage was evaluated in Osmanabadi breed bucks supplemented with organic Zinc (Zn) and Copper (Cu). A total number of 40 bucks aged 20 weeks were randomly assigned to 10 groups (each n = 4). The control group was maintained with basal diet, without any additional mineral supplementation. The treatment groups were supplemented with graded doses of organic Zn (Zn 20 mg, Zn 40 mg and Zn 60 mg), Cu (Cu 12.5 mg, Cu 25 mg and Cu 37.5 mg) and a combination of Zn + Cu (Zn 20 mg + Cu 12.5 mg, Zn 40 mg + Cu 25 mg and Zn 60 mg + Cu 37.5 mg), respectively for a period of 26 weeks (up to the age of 46 weeks). Sexual behaviour and scrotal biometry were recorded periodically. Blood and semen samples were collected and processed for LH estimation in blood plasma, and testosterone, T3 and T4 hormones in the seminal plasma. The mounts with ejaculation were observed earlier (P < 0.05) in the treatment bucks (from 38th week of age) than the control group (43rd week onwards). A positive correlation was observed between blood plasma LH and testosterone with total mounts (r = 0.31, P < 0.05; r = 0.51, P < 0.01) and mounts without ejaculation (r = 0.40, P < 0.01; r = 0.52, P < 0.01). A negative correlation between T4 with sperm number per ejaculation (r = −0.31, P < 0.05) and sperm concentration (r = −0.35, P < 0.05) had been noticed. Different doses of minerals showed positive interaction (P < 0.05) with sperm functional and behavioural characteristics. The spermatozoal gene expression of ODF2 and ZCCHC6 were significantly influenced by the mineral supplementation in all doses. The ZCCHC6 gene expression was positively correlated with testosterone (r = 0.50, P < 0.001) and sperm number per ejaculation (r = 0.42, P < 0.001), and ODF2 gene with T3 hormone (r = 0.34, P < 0.05). The present study indicates that the diet supplemented with organic trace minerals cause intense sexual behaviour, enhancement in sperm number per ejaculate, total motility, spermatozoal genes expression and altered LH, testosterone and T4 hormones. Graphical abstract Image 1 Highlights • First study regarding the role of Zn and Cu on male sexual behavior in goats. • Mineral supplementation influences LH, testosterone, T3 and T4 hormone levels. • Zn and Cu modulates the sperm quantitative and qualitative characteristics. • Mineral supplementation improved the male fertility genes (ODF2 and ZCCHC6) expression in spermatozoa. [ABSTRACT FROM AUTHOR]

Abstract The antioxidant properties and the protective role of organic zinc (Zn) and copper (Cu) in white blood cells (WBCs) and spermatozoa were analyzed through quantification of superoxide dismutase 1 (SOD1), catalase (CAT), glutathione peroxidase 4 (GPx4) and nuclear factor erythroid 2-like 2 (NFE2L2) and correlations were determined with sperm functional characteristics in Osmanabadi bucks. Bucks (aged 5 months; n = 40) were divided into ten groups, and the dietary treatments comprised of a control and nine treatment groups as follows: organic Zn as Zn 20, Zn 40 and Zn 60, organic Cu as Cu 12.5, Cu 25, Cu 37.5 and combined organic Zn and Cu as Zn 20+Cu 12.5, Zn 40+Cu 25, Zn 60+Cu 37.5, respectively per kg dry matter for a period of 8 months. The blood (120 and 240 days) and semen (240 days: 40 × 4 = 160) samples were collected from 40 bucks. In WBCs: the relative abundance of mRNA for SOD1, CAT, GPx4, NFE2L2 was greater (P < 0.05) in (120 and 240 days) in majority of the mineral supplemented animals. In spermatozoa: the relative abundance of SOD1, NFE2L2, GPx4 and CAT mRNA was greater (P < 0.05) in selected treatment groups. The abundance of SOD1 mRNA in WBCs was positively correlated (P < 0.05) with sperm mass motility (r = 0.692, P = 0.027). The abundance of GPx4 mRNA was negatively correlated (P < 0.05) with type A sperm (straightness; STR) > 85% and amplitude of lateral head displacement (ALH) > 2.5 μm/ s) (r = −0.711, P = 0.021) and (P < 0.05) positively correlated with sperm viability (r = 0.669, P = 0.035). Organic Zn and Cu supplementation was associated with an increase in the expression of antioxidant defense enzyme genes in bucks. [ABSTRACT FROM AUTHOR]

Abstract: The objective was to determine the association of mRNA expression of cystine rich secretary protein 2 (CRISP2), chaperonin containing T-complex protein 1, subunit 8 (CCT8), and phosphatidylethanolamine-binding protein 1 (PEBP1), in sperm of Holstein bulls with Sire Conception Rate (SCR) scores between −4 and +4. These proteins were involved in sperm capacitation and sperm-egg fusion. Samples of sperm obtained on a single day from Holstein bulls (N = 34) in a commercial AI centre were used to evaluate relative mRNA expression of CRISP2, CCT8, and PEBP1. The mRNA abundance of CRISP2 was positively correlated (r = 0.88; P < 0.002), CCT8 was negatively correlated (r = −0.87; P < 0.002), and PEBP1 was positively correlated (r = 0.83; P < 0.006) with SCR-scores. The means of CRISP2 mRNA abundance was greater among positive SCR-score bulls (2.5 to 8 fold), the means of CCT8 mRNA abundance was greater among the negative SCR-score bulls (9.5 to 3.5 fold), and the means of PEBP1 mRNA abundance was greater for the positive SCR-score bulls (5.4 to 7.7 fold). In multivariate regression models predicting SCR-scores, mRNA abundance of CCT8 was significantly associated with SCR-score in all models. In the presence of CRISP2 mRNA abundance in the model, the SCR score''s predictability of PEBP1 was insignificant. However, in the absence of CRISP2 mRNA abundance in the model, the SCR-score''s predictability of PEBP1 was significant. In multivariate regression models, CRISP2 and CCT8 mRNA expression in sperm accounted for 95% of the variance in Holstein bull''s SCR-scores. In conclusion, Holstein bulls with greater CRISP2 and lower CCT8 mRNA expression in sperm had higher probabilities of siring calves. [Copyright &y& Elsevier]

The high incidence of idiopathic recurrent pregnancy loss (iRPL) may stem from the limited research on male contributory factors. Many studies suggest that sperm DNA fragmentation and oxidative stress contribute to iRPL, but their roles are still debated. MicroRNAs (miRNAs) are short non-coding RNAs that regulate various biological processes by modulating gene expression. While differential expression of specific miRNAs has been observed in women suffering from recurrent miscarriages, paternal miRNAs remain unexplored. We hypothesize that analyzing sperm miRNAs can provide crucial insights into the pathophysiology of iRPL. Therefore, this study aims to identify dysregulated miRNAs in the spermatozoa of male partners of iRPL patients. Total mRNA was extracted from sperm samples of iRPL and control groups, followed by miRNA library preparation and high-output miRNA sequencing. Subsequently, raw sequence reads were processed for differential expression analysis, target prediction, and bioinformatics analysis. Twelve differentially expressed miRNAs were identified in the iRPL group, with eight miRNAs upregulated (hsa-miR-4454, hsa-miR-142-3p, hsa-miR-145-5p, hsa-miR-1290, hsa-miR-1246, hsa-miR-7977, hsa-miR-449c-5p, and hsa-miR-92b-3p) and four downregulated (hsa-miR-29c-3p, hsa-miR-30b-5p, hsa-miR-519a-2-5p, and hsa-miR-520b-5p). Functional enrichment analysis revealed that gene targets of the upregulated miRNAs are involved in various biological processes closely associated with sperm quality and embryonic development. [ABSTRACT FROM AUTHOR]

It is critical in sheep farming to accurately estimate ram fertility for maintaining reproductive effectiveness and for production profitability. However, there is currently a lack of reliable biomarkers to estimate semen quality and ramfertility, which is hindering advances in animal science and technology. The objective of this study was to uncover long non-coding RNAs (lncRNAs) in sperm from rams with distinct fertility phenotypes. Mature rams were allocated into two groups: high and low fertility (HF; n = 31; 94.5 ± 2.8%, LF; n = 25; 83.1 ± 5.73%; P = 0.028) according to the pregnancy rates sired by the rams (average pregnancy rate; 89.4 ± 7.2%). Total RNAs were isolated fromspermof the highest- and lowest-fertility rams (n = 4, pregnancy rate; 99.2 ± 1.6%, and 73.6 ± 4.4%, respectively) followed by next-generation sequencing of the transcripts. We uncovered 11,209 lncRNAs from the sperm of rams with HF and LF. In comparison to each other, there were 93 differentially expressed (DE) lncRNAs in sperm from the two distinct fertility phenotypes. Of these, 141 mRNAs were upregulated and 134 were downregulated between HF and LF, respectively. Genes commonly enriched for 9 + 2 motile cilium and sperm flagellum were ABHD2, AK1, CABS1, ROPN1, SEPTIN2, SLIRP, and TEKT3. Moreover, CABS1, CCDC39, CFAP97D1, ROPN1, SLIRP, TEKT3, and TTC12 were commonly enriched in flagellated sperm motility and sperm motility. Differentially expressed mRNAs were enriched in the top 16 KEGG pathways. Targets of the differentially expressed lncRNAs elucidate functions in cis and trans manner using the genetic context of the lncRNA locus, and lncRNA sequences revealed 471 mRNAs targets of 10 lncRNAs. This study illustrates the existence of potential lncRNA biomarkers that can be implemented in analyzing the quality of ram sperm and determining the sperm fertility and is used in breeding soundness exams for precision livestock farming to ensure food security on a global scale. [ABSTRACT FROM AUTHOR]

Male fertility can be affected by oxidative stress (OS), which occurs when an imbalance between the production of reactive oxygen species (ROS) and the body's ability to neutralize them arises. OS can damage cells and influence sperm production. High levels of lipid peroxidation have been linked to reduced sperm motility and decreased fertilization ability. This literature review discusses the most commonly used biomarkers to measure sperm damage caused by ROS, such as the high level of OS in seminal plasma as an indicator of imbalance in antioxidant activity. The investigated biomarkers include 8-hydroxy-2-deoxyguanosine acid (8-OHdG), a marker of DNA damage caused by ROS, and F2 isoprostanoids (8-isoprostanes) produced by lipid peroxidation. Furthermore, this review focuses on recent methodologies including the NGS polymorphisms and differentially expressed gene (DEG) analysis, as well as the epigenetic mechanisms linked to ROS during spermatogenesis along with new methodologies developed to evaluate OS biomarkers. Finally, this review addresses a valuable insight into the mechanisms of male infertility provided by these advances and how they have led to new treatment possibilities. Overall, the use of biomarkers to evaluate OS in male infertility has supplied innovative diagnostic and therapeutic approaches, enhancing our understanding of male infertility mechanisms. [ABSTRACT FROM AUTHOR]

Simple Summary: The objective was to identify candidate miRNAs and their integrated genes regulating acrosome function in capacitated sperm of high- versus low-fertility dairy bulls and to elucidate functional biological pathways using a systems biology approach featuring miRNA–mRNA cluster analyses. Based on categorized bovine miRNAs (n = 84), 19 were differentially expressed in high- compared to low-fertility capacitated sperm (p ≤ 0.05, fold regulation ≥ 2 magnitudes). mRNA expression of highly scored integrated genes of differentially expressed miRNAs was greater, ranging from 2.0 to 9.1-fold (p < 0.05) in high- compared to low-fertility sperm, with predicted pathways regulating acrosome vesicle exocytosis, acrosome reaction, and binding of sperm to zona pellucida. In conclusion, highly differentially expressed miRNAs in high-fertility bovine sperm regulating acrosome function have potential for predicting bull fertility. Bioinformatics envisage experimental data as illustrated biological networks, exploring roles of individual proteins and their interactions with other proteins in regulation of biological functions. The objective was to identify differentially expressed miRNAs and their associated genes regulating the acrosome reaction in capacitated sperm of high- compared to low-fertility dairy bulls and to elucidate biological functional pathways using a systems biology approach, featuring miRNA–mRNA cluster analysis. Categorized bovine-specific miRNAs (n = 84) were analyzed by RT-PCR; 19 were differentially expressed in high- compared to low-fertility sperm (p ≤ 0.05, fold regulation ≥ 2 magnitudes). Six miRNAs (bta-miR-129-5p, bta-miR-193a-3p, bta-miR-217, bta-mir-296-5p, bta-miR-27a, and bta-miR-320a) were highly upregulated (p < 0.05; fold regulation ≥ 5 magnitudes) in high- compared to low-fertility sperm. Highly scored integrated genes of differentially expressed miRNAs predicted associations with pathways regulating acrosome vesicle exocytosis, acrosome reaction, and sperm-oocyte binding. The mRNA expressions of genes associated with the acrosome reaction (including hub genes) were greater, ranging from 2.0 to 9.1-fold (p < 0.05) in high- compared to low-fertility capacitated bull sperm. In conclusion, differentially expressed miRNAs in high-fertility bovine sperm regulating acrosome functions have potential for predicting bull fertility. [ABSTRACT FROM AUTHOR]

Vechur is a unique but threatened indigenous cattle breed of Kerala. The use of cryopreserved semen serves long term conservation and improvement of the breed. The fertility of the cryopreserved sperm depends on the ability to withstand the freeze thaw cycle. Oxidative stress and reactive oxygen species (ROS) generated in the spermatozoa during freezing and thawing of frozen semen is considered a major factor responsible for causing infertility due to decrease in sperm quality parameters. Semen ejaculates from 20 Vechur bulls were collected. Bulls consistently producing semen with 70 per cent progressive motility were selected and analysed for cryotolerant sperm. Using conventional liquid Nitrogen method, six bulls exhibiting differential post thaw motility were grouped into Good Freezability Rate (GFR) (40 per cent) and Poor Freezability Rate (PFR) (<40 per cent) Three ejaculates from three bulls in each group were collected and cryopreserved. RNA was isolated from fresh and post thaw semen from two groups. Three candidate genes HSP90AA1, PRDX6 and SOD1 were selected for expression profiling. These genes were reported to be involved in the protection of the sperm from temperature changes, molecular folding (HSP90AA1) and oxidative stress (PRDX6 and SOD1). Relative quantification was performed by qPCR with PPIA as internal control gene. Statistical analysis revealed a significant (p<0.05) downregulation of HSP90AA1 and PRDX6 from fresh to GFR semen. While, there was no significant difference between fresh and GFR in case of SOD1 and a significant (p<0.05) downregulation in PFR from GFR for all the three selected candidate genes were revealed. The current study suggests the three candidate genes can be considered for selecting bulls based on sperm parameters. [ABSTRACT FROM AUTHOR]

Orange peel and its extract are good sources of phenols and vitamin C that can be used as powerful antioxidants and antibacterial. The effects of dietary ascorbic acid (AA), orange peel powder (OPP) and orange peel extract (OPE) supplementations on growth performance, blood biochemicals, gene expression and antioxidant status of growing rabbits under hot conditions were investigated. A total of 80 weaned Giant Flander male rabbits, five weeks old (606.25 ± 10.08 g), were randomly assigned to four groups. The first group received untreated diet (control group). The other groups received diets supplemented with 0.5 g AA/kg diet, 2% OPP and 500 mg OPE/kg diet. The lowest feed conversion ratio (FCR) was recorded by rabbits consumed diet supplemented with AA. Supplementations of OPP and OPE reduced blood plasma total cholesterol, low density lipoprotein and very-low density lipoprotein concentrations. The tested diets reduced triglycerides, total lipids, hydrogen peroxide, malondialdehyde levels, Staphylococcus aureus and Escherichia coli of the rabbits cecum. Supplementation of OPE improved activities of superoxide dismutase gene (6.1475) and insulin-like growth factor-1 (9.2108). Conclusively, dietary supplementation of OPE improved rabbit performance through improving antioxidant enzyme activities as well as upregulation of insulin-like growth gene. Additionally, OPP and OPE (2% and 500 mg/kg diet, respectively) had antibacterial effects for growing rabbits under hot conditions. [ABSTRACT FROM AUTHOR]

We evaluated the effects of feeding 60% dried corn distillers' grains plus solubles (DDGS) or the equivalent sulfur as CaSO4 on sperm characteristics and transcript abundance. Thirty-six half-sibling Angus bulls (256 ± 8.5 d; initial BW = 320 ± 2.7 kg) were assigned to one of three treatments: (1) 60% concentrate as corn (CON); (2) 60% DDGS as corn replacement (60DDGS); and (3) CON diet + equivalent sulfur of 60DDGS added as CaSO4 (SULF). The acrosome/cell membrane integrity, mitochondrial energy potential, oxidation status, DNA integrity, and zinc signatures were analyzed via flow cytometry. Sperm-specific gene expression was assessed via RNA sequencing. The flow cytometry data were analyzed using PROC MIXED in SAS to determine the effects of treatment. Pairwise comparisons based on edgeR were used to identify differentially expressed genes. The percentage of polarized mitochondria tended to be greater (p = 0.08) for SULF compared with CON and 60DDGS. Protamine 1 was upregulated (p < 0.01; FDR = 0.10) in 60DDGS compared to CON. Zinc signature 1 in 60DDGS and SULF was reduced (p = 0.03) compared to CON. This study suggests that feeding bulls diets containing 60% DDGS had little effect on DNA integrity and gene expression. [ABSTRACT FROM AUTHOR]

Background: MicroRNAs (miRNAs) are small, single-stranded, non-coding RNA molecules of 22–24 nucleotides that regulate gene expression. In the last decade, miRNAs have been described in sperm of several mammals, including cattle. It is known that miRNAs can act as key gene regulators of early embryogenesis in mice and humans; however, little is known about the content, expression, and function of sperm-borne miRNAs in early bovine embryo. In this study, total sperm RNA was isolated from 29 cryopreserved sperm samples (each coming from a separate bull) using a RNeasy kit and treatment with DNase I. RNA concentration and purity were determined through an Epoch spectrophotometer and an Agilent Bioanalyzer. The expression of 10 candidate miRNAs in bovine sperm (bta-miR-10a, bta-miR-10b, bta-miR-138, bta-miR-146b, bta-miR-19b, bta-miR-26a, bta-miR-34a, bta-miR-449a, bta-miR-495 and bta-miR-7), previously identified in testis and/or epididymis, was evaluated with RT-qPCR. The cel-miR-39-3p was used as a spike-in exogenous control. Nonparametric Mann–Whitney tests were run to evaluate which miRNAs were differentially expressed between bulls with high fertility [HF; non-return rates (NRR) ranging from 39.5 to 43.5] and those with subfertility (SF; NRR ranging from 33.3 to 39.3). Several sperm functionality parameters (e.g., viability, membrane stability or oxygen consumption, among others) were measured by multiplexing flow cytometry and oxygen sensing technologies. Results: RNA concentration and purity (260/280 nm ratio) (mean ± SD) from the 29 samples were 99.3 ± 84.6 ng/µL and 1.97 ± 0.72, respectively. Bioanalyzer results confirmed the lack of RNA from somatic cells. In terms of the presence or absence of miRNAs, and after applying the Livak method, 8 out of 10 miRNAs (bta-miR-10b, -138, -146b, -19b, -26a, -449a, -495, -7) were consistently detected in bovine sperm, whereas the other two (bta-miR-10a, and -34a) were absent. Interestingly, the relative expression of one miRNA (bta-miR-138) in sperm was significantly lower in the SF than in the HF group (P = 0.038). In addition to being associated to fertility potential, the presence of this miRNA was found to be negatively correlated with sperm oxygen consumption. The expression of three other miRNAs (bta-miR-19b, bta-miR-26a and bta-miR-7) was also correlated with sperm function variables. Conclusions: In conclusion, although functional validation studies are required to confirm these results, this study suggests that sperm bta-miR-138 is involved in fertilization events and beyond, and supports its use as a fertility biomarker in cattle. [ABSTRACT FROM AUTHOR]

The oriental river prawn (Macrobrachium nipponense) is a commercially important species in Asia. A previous study showed that the succinate dehydrogenase complex iron sulfur subunit B (SDHB) gene participates in testes development in this species through its effect on the expression of the insulin-like androgenic gland hormone gene. This study knocked-down the Mn-SDHB genes in M. nipponense using RNAi. A transcriptome analysis of the androgenic gland and testes was then performed to discover the male sex-related genes regulated by SDHB and investigate the mechanism of male sexual development in this species. More than 16,623 unigenes were discovered in each sample generated. In the androgenic gland, most of the differentially expressed genes were enriched in the hypertrophic cardiomyopathy pathway, while in the testes, they were enriched in the citrate cycle pathway. In addition, after Mn-SDHB knockdown, five genes were found to be downregulated in the androgenic gland in a series of biological processes associated with phosphorylated carbohydrate synthesis and transformations in the glycolysis/gluconeogenesis pathway. Moreover, a total of nine male sex-related genes were identified including Pro-resilin, insulin-like androgenic gland hormone, Protein mono-ADP-ribosyltransferase PAPR11, DNAJC2, C-type Lectin-1, Tyrosine-protein kinase Yes, Vigilin, and Sperm motility kinase Y-like, demonstrating the regulatory effects of Mn-SDHB, and providing a reference for the further study of the mechanisms of male development in M. nipponense. [ABSTRACT FROM AUTHOR]

Simple Summary: Bovine colostrum (BC) is the first secretion of mammary glands produced after birth; it represents a natural source of nutrients essential for the growth and development of newborns. Given its various properties, including antioxidants, this study intended to determine the effects of the dietary supplementation of two different concentrations (2.5% and 5%) of BC on the antioxidant status in different tissues (blood, liver and muscle) of rabbits. No differences in dietary treatment were recorded regarding the plasma concentration of antioxidant enzymes (catalase, glutathione peroxidase and superoxide dismutase), or regarding the gene expression of the aforementioned enzymes in the liver and longissimus dorsi (LD) muscle samples of rabbits. Further studies are needed to better understand the effect of this potentially promising nutraceutical in rabbit meat. Recent advances in animal nutrition have indicated that bovine colostrum (BC), due to its content of macronutrients, micronutrients and bioactive compounds, is an excellent health supplement. To the best of our knowledge, no studies on the effect of BC on antioxidant status have been performed in rabbits. This study aimed to investigate the effect of two BC concentrations on antioxidant status and gene expression of antioxidant enzymes in some tissues of rabbits. Thirty New Zealand White male rabbits were randomly divided into three experimental diets, containing 0% (CON), 2.5%, and 5% of BC (BC-2.5 and BC-5, respectively). The activity of antioxidant enzymes in plasma (catalase: CAT; glutathione peroxidase: GPx; superoxide dismutase: SOD), and the enzymes' gene expression in the liver and longissimus dorsi muscle, were determined. Results showed no significant differences, neither in plasma nor in tissues. A significant tissue-related effect has been observed regarding the mRNA levels of SOD and GPx, which were higher in the LD (p = 0.022) and liver (p = 0.001), respectively. Further studies, considering modifications of the length and dosage of dietary BC supplementation, are required to update the current state of knowledge in rabbits, as well as to fully understand the potential value of BC for possible application in farming use. [ABSTRACT FROM AUTHOR]

In bovines, cryopreserved semen is used for artificial insemination; however, the fertility of cryopreserved semen is far lower than that of fresh semen. Although cryopreservation alters sperm phenotypic characteristics, its effect on sperm molecular health is not thoroughly understood. The present study applied nextgeneration sequencing to investigate the effect of cryopreservation on the sperm transcriptomic composition of bull spermatozoa. While freshly ejaculated bull spermatozoa showed 14,280 transcripts, cryopreserved spermatozoa showed only 12,375 transcripts. Comparative analysis revealed that 241 genes were upregulated, 662 genes were downregulated, and 215 genes showed neutral expression in cryopreserved spermatozoa compared to fresh spermatozoa. Gene ontology analysis indicated that the dysregulated transcripts were involved in nucleic acid binding, transcriptionspecific activity, and protein kinase binding involving protein autophosphorylation, ventricular septum morphogenesis, and organ development. Moreover, the dysregulated genes in cryopreserved spermatozoa were involved in pathways associated with glycogen metabolism, MAPK signalling, embryonic organ morphogenesis, ectodermal placode formation, and regulation of protein auto-phosphorylation. These findings suggest that the cryopreservation process induced alterations in the abundance of sperm transcripts related to potential fertility-associated functions and pathways, which might partly explain the reduced fertility observed with cryopreserved bull spermatozoa. [ABSTRACT FROM AUTHOR]

Keywords: feedomics; gene expression; nutrigenomics; nutrition; transcriptome; ruminant EN feedomics gene expression nutrigenomics nutrition transcriptome ruminant N.PAG N.PAG 19 04/26/22 20220415 NES 220415 1. 10.1007/s00424-017-2105-9. 29305650 169 Arroyo J.M., Hosseini A., Zhou Z., Alharthi A., Trevisi E., Osorio J.S., Loor J.J. Reticulo-rumen mass, epithelium gene expression, and systemic biomarkers of metabolism and inflammation in Holstein dairy cows fed a high-energy diet. Transcription Factors (TFs) TFs are functional cellular proteins that manage the gene expression process through binding to target gene regulatory regions (silencer or promoter sequences) on the DNA, sparking gene expression series, and controlling the gene transcription rate [[39]]. 10.1038/s41598-018-35315-5 181 Liu L., Sun D., Mao S., Zhu W., Liu J. Infusion of sodium butyrate promotes rumen papillae growth and enhances expression of genes related to rumen epithelial VFA uptake and metabolism in neonatal twin lambs. This revolutionary approach has focused on how feeds talk to genes and how genes respond, addressing a novel holistic approach and redefining the conventional ruminant nutrition-gene pattern in a broad context. [Extracted from the article]

Despite encountering new challenges in using epididymal sperm recovered from cauda epididymides, this accessible and, in some species, worthwhile sample makes inevitable the further development of a suitable cryopreservation protocol. In this study, sperm was recovered from the epididymis of 4°C overnight stored slaughtered bulls' testes and the effects of cryopreservation on the bovine epididymal sperm motility (with CASA) and gene expression patterns (with quantitative Real time‐PCR) were evaluated. Moreover the fertilizing potential of cryopreserved epididymal sperm was used in in vitro fertilization (IVF). After freezing and thawing of epididymal sperm, total and slow progressive sperm motility, VCL, VAP, MAD, ALH and BCF were significantly decreased (p <.05), while in the parameters of fast progressive motility, VSL, LIN, WOB and STR there were not any significant variations in the frozen sperm compared to fresh (non‐frozen) counterpart. The assessment of abundance of transcripts encoding motility (TSSK6) and fertility (PRM1 and PRM2)‐related genes in epididymal sperm, showed that these transcripts were affected by freezing especially in slow progressive motility status (p <.01). Furthermore, cleavage and blastocyst rate did not present any significant differences between bovine embryos produced in vitro by fresh or frozen‐thawed epididymal sperm. It can be concluded that epididymal sperm has enough freezability after overnight testes storage, and cryopreservation could not affect the percentage of in vitro produced embryos in spite of the changes of relative abundance of some transcripts and direction progressive motility pattern of sperm. [ABSTRACT FROM AUTHOR]

In this study, chrysin as a natural flavonoid was encapsulated in nanoliposomal structures, and the synthesized nanoliposome-loaded chrysin (NLC) was further characterized for its physical properties and cytoprotective effects in mice that received cadmium-containing water. The results showed that the synthesized NLC is possessed spherical structure with the size of 185.1 nm and negative surface charge of − 26 mV with a poly dispersity index of 0.26. The mice received cadmium (2 mg/kg body weight/day) through drinking water showed weight loss and decease in the feed intake significantly (p ≤ 0.05). The cadmium notably (p ≤ 0.05) increased the liver enzymes including aspartate aminotransferase, alanine transaminase, and alkaline phosphatase; altered the liver metal deposition (cadmium, copper, manganese, selenium, and zinc); and induced hepatic oxidative stress (inducible nitric oxide synthase, catalase, superoxide dismutase, and glutathione peroxidase genes) with no remarkable histopathological changes. Furthermore, the cadmium impaired the morphology of jejunum through reducing villus height and villus width and increasing the crypt depth. Providing NLC as a dietary supplement at the concentrations of 2.5 and 5 mg/kg mice body weight significantly (p ≤ 0.05) improved the feed intake and body weight gain, modulated the liver enzymes, and alleviated the hepatic oxidative stress. The NLC also improved the antioxidant mineral deposition in the liver and morphohistological structure of jejunum. Consequently, the NLC is suggested as a potential dietary supplement to alleviate the symptoms of cadmium-induced toxicity in mice. [ABSTRACT FROM AUTHOR]

Background: One of the important factor associated with male infertility is high production of reactive oxygen species (ROS). The main function of Nuclear factor erythroid 2-related factor 2 (NRF2) is to activate the cellular antioxidant response by inducing the transcription of a wide array of genes that can combat the harmful effects of factors such as oxidative stress. The purpose of this study was to evaluate the effect of N-acetyl-L-cysteine (NAC), as an antioxidant drug, on NRF2 Gene Expression in Asthenoteratozoospermia Men. Materials and Methods: In this randomized, blinded clinical trial study, included 50 infertile men with asthenoteratozoospermia, who received NAC (600 mg, three times daily). Sperm parameters analyzed according to the world health organization (WHO; 2010). Sperm DNA fragmentation, relative NRF2 expression, and seminal plasma level of antioxidant enzymes were measured by TUNEL assay, reverse transcription polymerase chain reaction (RT-PCR) and ELISA test, respectively. Results: After NAC treatment, findings showed a significant increase in sperm concentration and motility compared to pre-treatment status, whereas the percentage of abnormal morphology and DNA fragmentation was significantly decreased (P<0.05). A significant improvement in expression of NRF2 gene and antioxidant enzyme levels were observed compared to pre-treatment by NAC (P<0.05). Significant correlations were observed between NRF2 mRNA expression level, specific sperm parameters and level of antioxidant enzymes (P<0.05). Conclusion: The results demonstrated that NAC oral supplementation protected against oxidative stress by enhancing NRF2 expression. This could improve semen parameters quality parameters in asthenoteratozoospermia men [ABSTRACT FROM AUTHOR]

Summary: Predicting bull fertility prior to breeding is a current challenge for the dairy industry. The use of molecular biomarkers has been previously assessed. However, the integration of this information has not been performed to extract biologically relevant markers. The goal of this study was to integrate DNA methylation data with previously published RNA‐sequencing results in order to identify candidate markers for sire fertility. A total of 1765 differentially methylated cytosines were found between high‐ and low‐fertility sires. Ten genes associated with 11 differentially methylated cytosines were found in a previous study of gene expression between high‐ and low‐fertility sires. Additionally, two of these genes code for proteins found exclusively in bull seminal plasma. Collectively, our results reveal 10 genes that could be used in the future as a panel for predicting bull fertility. [ABSTRACT FROM AUTHOR]

Background and aims: miRNAs have regulatory functions in developmental processes. The stages of spermatogenesis can also be affected by miRNAs. We tried to detect a relationship between the expression of miR-508-5p and miR-510-5p and male infertility. The purpose of this study was to investigate the association between expression of miR-508-5p and mir-510-5p in ejaculated sperm from patients with idiopathic asthenozoospermia, teratozoospermia, and teratoasthenozoospermia. Materials and Methods: We enrolled 18 men with asthenozoospermia, 17 men with teratozoospermia, 18 men with teratoasthenozoospermia, and 18 individuals with normozoospermia based on the clinical criteria. Then, the expressions of the mentioned miRs in the spermatozoa were evaluated by quantitative real-time PCR (qRT-PCR). Kruskal-Wallis was used to compare their expressions in the studied groups. Results: The expression of mir-508-5p did not show any statistical significance in all groups. On the other hand, the expression of miR-510- 5p in teratozoospermia groups (P<0.05) and the asthenozoospermia group (P<0.05) demonstrated a significant downregulation compared with the control and teratoasthenozoospermia groups. Conclusion: By analyzing the expression profile of miRNAs, we concluded that the expression level of miR-510-5p changed in patients with abnormal motility and morphology of spermatozoa; therefore, it may affect infertility by down-regulating the expression of mir-510-5p which shows the role of it in abnormal morphology and motility defects in infertility cases. [ABSTRACT FROM AUTHOR]

An introduction is presented in which the author discusses topics within the issue including gene expression, monoclonal antibodies, and chromatin.

Conventional sperm analysis (i.e., motility and fertility) has been used to evaluate sperm quality. Understanding the quality of sperm on the molecular level in the sturgeons, Acipenser baerii and A. schrenckii, is essential for the improvement of the conservation of genetic resources and farming performance. In this study, we used the iTRAQ proteomics approach to perform proteomic profiling of spermatozoa associated with sperm quality in sturgeons (Data are available via ProteomeXchange with identifier PXD006108). The results showed 291 and 359 differentially expressed proteins in A. baerii and A. schrenckii, respectively, of which 72 were common to both species and all were upregulated in high quality compared with low quality samples. The differentially expressed proteins were mainly categorized into the generation of precursor metabolites and energy and oxidation, and they were localized to the mitochondria. Three distinguishing pathways, Arginine and proline metabolism, Pyruvate metabolism and the Citrate cycle (TCA cycle) were found to play an important role in energy metabolism, and some substrates could be used in the sperm medium for storage and cryopreservation. The quantity levels of two proteins, CKMT1 and LDHB, were verified by western blot analysis. Moreover, other potential biomarkers involved in oxidation reduction, ubiquitin-proteasome-dependent proteolysis, chaperones and binding activity were also discussed. Our study is the first to use the iTRAQ-based proteomics approach to analyse the sturgeon spermatozoa proteome, and the results that we obtained are valuable for the prediction of sperm quality and reproduction management in these threatened species. [ABSTRACT FROM AUTHOR]

Spermatozoal transcripts expression levels could be used to assess fertility potential of a male. The objective of the present study was to elucidate the predictive ability of the expression levels of growth, apoptosis and homeostasis regulating transcripts on sperm functions and fertility. The expression levels of spermatozoal RNA isolated from the neat semen samples were related to the good (discarded ejaculate, <25%; n = 7) and poor (discarded ejaculate, >40%, n = 6) quality semen producer and bulls ( n = 12) with known conception rate. The relative fold expression levels of BMP2 were significantly ( p < 0.01) higher in good than the poor semen producers and positively associated with post-thaw sperm velocity parameters (LIN and VAP). The NGF expressions fold levels had significant ( p < 0.05) positive relationship with mitochondrial membrane potential of neat semen samples. The genes involved in the apoptotic, UBE2D3 ( r = −0.61, p = 0.02), CASP3 ( r = −0.57, p = 0.03) and homeostatic, HSFY2 ( r = −0.61, p < 0.02) regulators had significant negative correlation with the percentage of post-thaw fast progressive motile spermatozoa. The expression level of TRADD had significant negative influence on the mitochondrial membrane potential ( r = −0.54, p = 0.05) of neat semen samples and conception rate ( r = −0.57, p < 0.05). The expression levels of BMP2 had highly significant positive correlation with NGF ( r = 0.99, p < 0.01) and CASP3 ( r = 0.56, p = 0.05). The BMP2 expression level might be used to predict the quality of the semen and TRADD determine the conception rate of the bull. The study provides ample evidence that the sperm transcripts expression levels might be used to predict quality semen production and bull fertility. [ABSTRACT FROM AUTHOR]

In the current study, fennel essential oil was used as an antibiotic alternative compared to gentamycin for enhancing the expression of apoptosis genes and antioxidant enzymes in weaned rabbits as well as meat quality and growth performance. The gene expression of the cell lymphoma 2 (BAX and BCL2), caspase3 (CASP3), and glutathione peroxidase (GPX1) were estimated in the liver tissue using qRT-PCR. A total of 45 Moshtohor weaned male rabbits aged four weeks were randomly allocated to control, T1, and T2 treatment groups; each consisted of 15 weaned male rabbits with five replicates. Rabbits in the T1 and T2 groups were orally supplied with 1 mL fennel oil and 1 mL gentamycin, respectively. Weaned rabbits under different treatments showed increased body weight (BW) at 8 and 12 weeks of age and average daily gain (ADG) at 4–8 and 4–12 weeks of age compared to the control group. Compared to the controls, the weaned rabbits supplemented with fennel oil and gentamycin had lower total cholesterol, triglyceride, and MDA. In addition, villus length, mRNA of BAX, BCL2, Casp3, and GPX were increased in the different treatments compared to the control. Furthermore, the meat of these rabbits was less tender, had a lower aerobic plate count (APC), pH, and was brighter and redder in color than the control. Under the conditions of the present study, the supplementation of weaned Moshtohor rabbits with fennel oil as a natural alternative for gentamycin enhanced feed conversion and daily gain through enhancing villus length and mucus thickness. Additionally, fennel essential oil reduces oxidative stress by increasing the antioxidant enzymes. [ABSTRACT FROM AUTHOR]

Standard sperm evaluation parameters do not enable predicting their ability to survive cryopreservation. Mitochondria are highly prone to suffer injuries during freezing, and any abnormalities in their morphology or function are reflected by a decline of sperm quality. Our work focused on describing a link between the number and the activity of mitochondria, with an aim to validate its applicability as a biomarker of bovine sperm quality. Cryopreserved sperm collected from bulls with high (group 1) and low (group 2) semen quality was separated by swim up. The spermatozoa of group 1 overall retained more mitochondria (MitoTrackerGreen) and mtDNA copies, irrespective of the fraction. Regardless of the initial ejaculate quality, the motile sperm contained significantly more mitochondria and mtDNA copies. The same trend was observed for mitochondrial membrane potential (ΔΨm, JC-1), where motile sperm displayed high ΔΨm. These results stay in agreement with transcript-level evaluation (real-time polymerase chain reaction, PCR) of antioxidant enzymes (PRDX1, SOD1, GSS), which protect cells from the reactive oxygen species. An overall higher level of glutathione synthetase (GSS) mRNA was noted in group 1 bulls, suggesting higher ability to counteract free radicals. No differences were noted between basal oxygen consumption rate (OCR) (Seahorse XF Agilent) and ATP-linked respiration for group 1 and 2 bulls. In conclusion, mitochondrial content and activity may be used as reliable markers for bovine sperm quality evaluation. [ABSTRACT FROM AUTHOR]

Seminal plasma (SP), the non-cellular component of semen, is a heterogeneous composite fluid built by secretions of the testis, the epididymis and the accessory sexual glands. Its composition, despite species-specific anatomical peculiarities, consistently contains inorganic ions, specific hormones, proteins and peptides, including cytokines and enzymes, cholesterol, DNA and RNA—the latter often protected within epididymis- or prostate-derived extracellular vesicles. It is beyond question that the SP participates in diverse aspects of sperm function pre-fertilization events. The SP also interacts with the various compartments of the tubular genital tract, triggering changes in gene function that prepares for an eventual successful pregnancy; thus, it ultimately modulates fertility. Despite these concepts, it is imperative to remember that SP-free spermatozoa (epididymal or washed ejaculated) are still fertile, so this review shall focus on the differences between the in vivo roles of the SP following semen deposition in the female and those regarding additions of SP on spermatozoa handled for artificial reproduction, including cryopreservation, from artificial insemination to in vitro fertilization. This review attempts, including our own results on model animal species, to critically summarize the current knowledge of the reproductive roles played by SP components, particularly in our own species, which is increasingly affected by infertility. The ultimate goal is to reconcile the delicate balance between the SP molecular concentration and their concerted effects after temporal exposure in vivo. We aim to appraise the functions of the SP components, their relevance as diagnostic biomarkers and their value as eventual additives to refine reproductive strategies, including biotechnologies, in livestock models and humans. [ABSTRACT FROM AUTHOR]