Selenium–curcumin conjugated nanoparticles (curconSeNPs) were synthesized using an ascorbic acid–mediated reduction method to enhance the biofunctional stability of selenium and curcumin nanoforms. Selenium, an essential antioxidant micronutrient, attains improved bioavailability and reduced reactivity at the nanoscale, and its conjugation with curcumin further strengthens these advantages. The synthesized selenium nanoparticles (SeNPs), curcumin nanoparticles (CurNPs), and curconSeNPs were characterized for size distribution, morphology, elemental composition, surface charge, and functional groups. Conjugation reduced the hydrodynamic particle size from 71 nm (SeNPs) to 54 nm, while the optical band gap decreased from 2.13 eV to 1.61 eV, indicating altered electronic properties. SEM, TEM, and SAED analyses confirmed spherical nanoparticles with a rhombohedral crystal structure, and EDAX revealed clear signatures of selenium and oxygen. The lattice fringe spacing measured 5 nm for SeNPs and 10 nm for curconSeNPs. FTIR spectra verified the presence of curcumin and selenium functional groups, and SAED revealed additional biofunctional groups associated with the conjugate. CurconSeNPs exhibited enhanced colloidal stability, with a zeta potential of -35.6 mV compared with − 33 mV for SeNPs and − 12.3 mV for CurNPs. Acute oral toxicity studies in Wistar rats demonstrated a wide safety margin for all formulations, indicating their suitability for future biotherapeutic and nutraceutical applications.

Spermatogonial stem cells (SSC) are of high significance in animal reproduction, breeding, and regenerative medicine. Cryopreservation of putative SSC is a prerequisite for long-term storage and future applications. However, dissociated putative SSC are very susceptible to cryostress and undergo apoptosis, thereby reducing their functional competence. To minimize the dissociation-induced apoptosis during cryopreservation and post-thaw culture, the role of anti-apoptotic molecule, Y-27632, was evaluated in sheep putative SSC. The putative SSC were isolated, enriched, and cultured from prepubertal sheep testis. Y-27632 (0, 5,10, and 15 µM) was added in putative SSC cryomedia (Cy group) and in both freezing and post-thaw culture media (Cy + y group). The effects were compared with non-treated control (C group). One month after cryopreservation, the putative SSC were assessed for viability, ROS production, metabolic activity, stemness associated undifferentiated status, colony characteristics, and apoptosis associated pathway/genes. The viability of the dissociated post-thaw putative SSC was significantly higher > 90

Sperm cryopreservation is a critical component of assisted reproductive technologies employed for both livestock breeding and human fertility management. Sperm are the highly specialized motile cells prone to cryodamage during freezing. Moreover, buffalo, pig and sheep sperm are more susceptible to cryoinjury leading to increased semen rejection rates and substantial economic losses due to reduced fertility. Advances in freezing protocols and modulation in composition of semen diluents protect sperm from cryodamage; however, inconsistency and inter-individual variability in semen freezability exist due to multifactorial etiology. The use of molecular technologies, particularly genomics, transcriptomics, proteomics, and metabolomics led to identification of potential biomarkers associated with cryotolerance. These omics-driven insights have not only enlightened our understanding of the molecular basis of cryoinjury but also has the potential in selecting bulls with good semen freezability. A multidisciplinary approach toward the development of targeted strategies such as supplementing extenders with novel cryotolerant biomolecules to mitigate the sperm damage. This review consolidates current knowledge on the molecular and physiological underpinnings of sperm cryodamage offering a holistic perspective that may guide refinement of existing cryopreservation protocols and extenders for improving sperm cryo-survivability in breeding males.

The spermatogonial stem cells (SSCs) are unique stem cells of the adult body having the power of self-renewal and differ entiation into spermatozoa. Due to their distinct advantages over other stem cell types, the field of cell based regenerative medicine is exploring the potential of SSCs for its various applications in biotechnology and animal reproduction. Here we discuss about several approaches for SSC isolation, enrichment and characterization. Given the low percentage of SSCs in the testis, it is difficult to obtain an enriched population of SSC for further research. Differentiation and apoptosis over time is another challenge in establishing a long term SSC culture system. Hence, optimization of the culture media and culture conditions along with cryopreservation is a prerequisite for the downstream application of SSC technology in livestock. Although several markers have been identified for SSCs in animals, an appropriate species specific marker is still lacking. Suitable recipients for SSC transplantation are generated through surrogate sire technology in livestock. SSC transplantation has been experimentally successful in numerous livestock species with donor derived spermatogenesis and production of offspring. Such advancement coupled with genome editing may open new avenues for augmenting livestock production in near future. We emphasize that a deeper understanding of the SSCs characteristics and the fac tors that influence their differentiation and stemness is essential for the wider application of SSC technology in animals.

Turmeric, a bright yellow spice from India, belongs to the Curcuma family. The phytochemical compounds curcumin, demethoxycurcumin and bisdemethoxycurcumin (curcuminoids) in turmeric are traditionally used as an antimicrobial agent and also as a growth enhancer in both livestock and poultry. Among these phytocompounds in turmeric, the lipophilic polyphenol compound, curcumin (1,7-bis-(4-hydroxy-3-methoxyphenyl)-hepta-1,6-diene-3,5-dione), has unique properties such as anti-bacterial, anti-inflammatory, anti-oxidant, and anti-cancer effects. However, curcumin also has low water solubility, bioavailability and pharmacokinetic properties for therapeutic applications. To overcome these drawbacks, a variety of curcumin formulations have been established. Recently, nanotechnology emerged as vibrant subject to address variety of issues including enhancing the bioavailability. The synthesized nanocurcumin could be employed as an animal feed additive to improve the reproductive health, egg and meat quality, gut health and growth promotion. The synthesized nanocurcumin can be analyzed by various analytical techniques such as UV–Visible spectroscopy, FTIR and HPLC analysis. In addition, different extraction methods are involved; its formulations, identification and characterization are essential for its application in the animal feed and for enhanced biological activities.

Nanoparticles have been widely synthesized and used in various biological applications. Among the minerals, zinc (Zn) alters numerous biological pathways, and curcumin (Cur) functions mainly as the antimicrobial, anti-inflammatory, and anticancerous agent. In this study, nano-zinc oxide (nZnO), nano-curcumin (nCur), and its conjugate (nZnOcur) were synthesized through the chemical reduction method. The particles were characterized using UV-visible (UV-visible) spectroscopy, Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), scanning electron microscopy-energy-dispersive X-ray spectroscopy (SEM-EDS), high-resolution transmission electron microscopy (HR-TEM), dynamic light scattering (DLS), and ζ-potential measurements. The UV-visible spectrum revealed that nZnO had an absorbance peak at 370 nm, while nCur had peaks at 400-600 nm, and the nZnOcur conjugate had peaks at 550-650 nm and a smaller peak in the 300-400 nm range. The key functional groups identified by FTIR confirm the successful conjugation of nZnOcur nanoparticles. XRD patterns indicated the crystalline wurtzite phase of nZnO with variations in peak intensity, suggesting structural changes upon curcumin incorporation. SEM-EDS and DLS analyses provided average particle sizes of 88.41 and 122 nm; 53.81 and 98 nm; 43.41 and 102 nm for nZnO, nCur, and nZnOcur conjugate, respectively. HR-TEM images showed that nZnO (∼70 nm, spherical) transformed into rod-like nZnOcur (∼105 nm) upon conjugation with curcumin. nZnOcur had higher antibacterial activity against bacterial strains, demonstrating synergistic effects. The antioxidant capacity was greater in both nCur and nZnOcur conjugates, while anti-inflammatory assays indicated significant protein denaturation inhibition with all three nanoparticles. Cytotoxicity tests revealed that nZnO had higher toxicity, and the acute oral toxicity study indicated nZnOcur conjugate as least toxic, with LD50 value of 425.4 mg/kg. Overall, the findings underscore the nZnOcur conjugate as a promising and safer nanoparticle for antimicrobial, anti-inflammatory, and antioxidant applications with least side effects.

The application of spermatogonial stem cells (SSC) will be more effective and feasible following the successful cryopreservation and transfer of SSCs in livestock. Like other cells, SSCs are also sensitive to cryoinjury; hence composition of the cryomedia and freezing protocols need to be optimized. The present study aims to optimize the best freezing rates by minimising the ice crystallization and dehydration effect in order to maximize the post-thaw SSCs survivability and stemness characteristics. Three different freezing protocols with varied cooling profiles, cooling profile 1 (isopropanol based freezing): 1 °C/min from 0 °C to -10 °C, 0.5 °C/min up to -40 °C, further reduced to 0.25 °C/min up to -50 °C and 0.1 °C/min to -60 °C; cooling profile 2 (using programmable freezer): 1 °C/min up to 4 °C, 0.3 °C/min up to -8 °C, and cooled at 0.5 °C/min to -50 °C, further decrease to -90 °C (8 °C/min) and cooling profile 3 (uncontrolled rapid freezing): 3.3 °C/min from 0 °C to -10 °C, 5 °C/min up to -40 °C, 2 °C/min to -50 °C and 1.2 °C/min up to -60 °C, were compared for cryopreservation efficiency. The overall viability (91.41 ± 2.00 % Vs 74.59 ± 2.34 %), stemness activity (1.34 ± 0.095 OD units Vs 0.356 ± 0.026 OD units), and proliferation rate (0.849 ± 0.019 OD units Vs 0.749 ± 0.015 OD units) of post-thaw SSC culture irrespective of the freezing regimes were significantly decreased when compared to pre-freeze SSC culture characteristics. The post-thaw viability was significantly greater in cooling profile 1 (79.64 ± 4.1 %) when compared to cooling profile 2 (69.72 ± 2.4 %) and cooling profile 3 (75.43 ± 4.8 %). Also, cooling profile 1 yielded greater (p < 0.05) post-thaw stemness activity (0.456 ± 0.044 OD units) when compared to other methods. The study suggests that the cooling profile 1 using isopropanol based freezing can be recommended for preservation of viability and stemness characteristics of SSCs.

The present study evaluated the effect of dietary supplementation with n-3 polyunsaturated fatty acids (PUFA) on preovulatory follicle (POF) turnover, prolificacy, and endocrine and metabolic milieu in Malpura sheep. Fifty cyclic ewes with 3-3.5 body condition scores on a five-point scale were allocated equally to two groups (n = 25) following estrus synchronization and were supplemented with 0.6 mL/kg body weight of n-3 PUFA-rich fish oil (FO) or palm oil (PO) as control, for 60 d following an acclimatization period of 7 d. All ewes were mated with sexually active rams at the end of the supplementation period. On ultrasonographic ovarian scanning at the last fourth estrus, the mean number of POFs was 77.8% greater (P < 0.01) in FO ewes than in the PO ewes. The proportion of ewes with multiple ovulations two months after the beginning of supplementation was 56% in the FO group as compared to 8% in the PO group. The number of fetuses was 46% higher (P < 0.01) in the FO than in the PO ewes at d 45 of gestation. At lambing, the twinning percent in the FO ewes was three times greater than in the PO ewes (27.3 vs. 9.1%). Plasma cholesterol, estradiol, and insulin concentrations were lower (P < 0.01) in ewes fed with FO than those offered PO group at the end of the feeding period. It was concluded that the dietary supplementation of n-3 PUFA-rich FO in well-fed Malpura ewes improved the number of follicles and ovulation rate which led to an increased prolificacy, accompanied by a reduction of plasma cholesterols, estradiol, and insulin.

Sperm transcriptomics provide insights into subtle differences in sperm fertilization competence. For predicting the success of complex traits like male fertility, identification of hub genes involved in various sperm functions are essential. The bulls from the transcriptome profiled samples (n = 21), were grouped into good and poor progressive motility (PM), acrosome integrity (AI), functional membrane integrity (FMI) and fertility rate (FR) groups. The up-regulated genes identified in each group were 87, 470, 1715 and 36, respectively. Gene networks were constructed using up- and down-regulated genes from each group. The top clusters from the upregulated gene networks of the PM, AI, FMI and FR groups were involved in tyrosine kinase (FDR = 1.61E−11), apoptosis (FDR = 1.65E−8), translation (FDR = 2.2E−16) and ribosomal pathway (FDR = 1.98E−21), respectively. From the clusters, the hub genes were identified and validated in a fresh set of semen samples (n = 12) using RT-qPCR. Importantly, the genes (fold change) RPL36AL (14.99) in AI, EIF5A (54.32) in FMI, and RPLP0 (8.55) and RPS28 (13.42) in FR were significantly (p < 0.05) up-regulated. The study suggests that the expression levels of MAPK3 (PM), RPL36AL + RPS27A or RPL36AL + EXT2 (AI), RPL36AL or RPS27A (FMI) and RPS18 + RPS28 (FR) are potential markers for diagnosing the semen quality and fertility status of bulls which can be used for the breeding program.

Sperm antigenicity has been implicated as a regulatory factor for acquiring fertilizing competence in the female reproductive tract. Overt immune response against the sperm proteins leads to idiopathic infertility. Hence, the aim of the study was to evaluate the influence of the auto-antigenic potential of sperm on the antioxidant status, metabolic activities and reactive oxygen species (ROS) in bovine. Semen from Holstein-Friesian bulls (n = 15) was collected and classified into higher (HA, n = 8) and lower (LA, n = 7) antigenic groups based on micro-titer agglutination assay. The neat semen was subjected to the evaluation of bacterial load, leukocyte count, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and lipid peroxidation (LPO) levels. Antioxidant activities in seminal plasma and intracellular ROS levels in the post-thawed sperm were estimated. The number of leukocytes was lower (p < .05) in the HA than the LA semen. The percentage of metabolically active sperm was higher (p < .05) in HA than the LA group. The activities of total non-enzymatic antioxidant, superoxide dismutase (SOD) and catalase (CAT) were higher (p < .05) while glutathione peroxidase activity was lower (p < .05) in the seminal plasma of LA group. The LPO levels of neat sperm and the percentage sperm positive for intracellular ROS in the cryopreserved sample were lower (p < .05) in the HA group. Auto-antigenic levels were positively correlated with the percentage of metabolically active sperm (r = 0.73, p < .01). However, the seminal auto-antigenicity was negatively (p < .05) correlated with the levels of SOD (r=-0.66), CAT (r=-0.72), LPO (r=-0.602) and intracellular ROS (r=-0.835). The findings were represented in graphical abstract. It is inferred that the higher auto-antigenic levels protect the quality of bovine semen by promoting sperm metabolism and lowering ROS and LPO levels.

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 up-regulated 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.

The study aimed to assess the influence of cryostress on RNA integrity and functional significance in sperm fertilizing ability. The fresh and post-thawed buffalo sperm ( n = 6 each) samples were evaluated for their functional attributes, and sperm total RNA was subjected to transcriptome sequencing followed by validation using real-time PCR and dot blot. Overall, 6911 genes had an expression of FPKM > 1, and among these 431 genes were abundantly expressed (FPKM > 20) in buffalo sperm. These abundantly expressed genes regulate reproductive functions such as sperm motility ( TEKT2 , SPEM1 , and PRM3 , FDR = 1.10E-08), fertilization ( EQTN , PLCZ1 , and SPESP1 , FDR = 7.25E-06) and the developmental process involved in reproduction ( SPACA1 , TNP1 , and YBX2 , FDR = 7.21E-06). Cryopreservation significantly ( p < 0.05) affected the structural and functional membrane integrities of sperm. The expression levels of transcripts that regulate the metabolic activities and fertility-related functions were compromised during cryopreservation. Interestingly, cryostress induces the expression of genes involved ( p < 0.05) in chemokine signaling ( CX3CL1 , CCL20 , and CXCR4 ), G-protein coupled receptor binding ( ADRB1 , EDN1, and BRS3 ), translation ( RPS28 , MRPL28 , and RPL18A ), oxidative phosphorylation ( ND1 , ND2 , and COX2 ), response to reactive oxygen species ( GLRX2 , HYAL2 , and EDN1 ), and immune responses ( CX3CL1 , CCL26 , and TBXA2R ). These precociously expressed genes during cryopreservation alter the signaling mechanisms that govern sperm functional competence and can impact fertilization and early embryonic development.

This study aimed to investigate the influence of calcium and magnesium supplementation on the skewing of sex ratio and placental genes expression in New Zealand White Rabbits. A total of 25 rabbits were allocated to 5 groups; each treatment group was supplemented with Ca and Mg; T1 (0.4% and 0.01%), T2 (0.6% and 0.02%), T3 (0.8% and 0.03%), and T4 (1.0% and 0.04%) respectively, while the C group was provided with regular feed and subjected to three breeding. The T3 group produced a female-biased litter and hence, the F1 female kits (n=12) of T3 and C group was provided same supplementation management. At 21 days of gestation, three animals from each group were sacrificed and placental samples were collected, the remaining animals were allowed for full-term delivery. The selected F1 produced female-biased litter with elevated serum Ca and Mg concentrations and reduced sodium and cholesterol levels. A total of 15 genes related to mineral absorption, placental development and immunity were selected to study the influence of diet on sex and placental gene expression. The expression of genes such as PEG10, SOD1, SLC30A, TLR4, AR, and TRPM6 was high in the treatment placenta compared to the control. RTL1, ESR2, CALM2, and TRPM6 upregulated in the treatment female placenta. The study concluded that the intake of 0.8% Ca and 0.03% Mg could lead to the production of more females and upregulation of some placental genes could serve as the molecular mechanism in response to intake of the minerals and fetal sex.

Background: Oxidative stress negatively impacts pregnancy and its outcomes; hence, its avoidance through gestational antioxidant intakes is a desired nutrition-reproduction practices. Materials and Methods: Female rabbits (n=40) were divided into 5 groups and supplemented with 0, 200, 300, 400, and 500 mg Chlorella vulgaris biomass per kilogram body weight daily, respectively throughout the gestation period. Upon kindling, kits of the female rabbits (n=75) were randomly selected and monitored from birth till 120 days old. The reproductive performance of the female rabbits was evaluated by computation of their reproductive index, while birth weight, growth rate, oestrogen and testosterone concentrations, oxidative stress biomarkers, and expression of selected functional genes of the progeny were determined. Results: The gestational intake of Chlorella vulgaris increased reproductive index of the female rabbits (p < 0.05), and it also increases the growth and oxidative stress protection status of the rabbit progeny (p < 0.05). Selected functional genes, including Gstp1, Cyp1a1, Ar, Ghr, Il2, and Il6 assessed in the progeny of the supplemented groups were significantly upregulated (p < 0.05). Conclusion: It was concluded from these results that gestational Chlorella vulgaris biomass intake improved reproductive index of the female rabbits, increased growth and oxidative stress protection in their progeny

Energy based diet supplementation is responsible for body growth and plays a significant role to improve spermatogenesis, semen quality, fertility and influence livestock production. The aim of the present study was to evaluate the effect of dietary flaxseed and coconut oil supplementation on semen profile, hormone level, testicular biometry, antioxidant enzyme activity and seminal plasma protein in adult rams. A total number of nine rams (Bannur crossbred rams, aged 20-23 months) were divided into three groups and each group consisted of three rams. The sheep of the control were fed a diet consisting of concentrate mixture and ragi straw in a ratio of 50:50, respectively as per ICAR 2013 standard. The sheep of the coconut oil supplemented group were fed the same basal diet as that of the control group; additionally they were supplemented with coconut oil @ 5 g/100 g concentrate mixture. The sheep of flaxseed oil supplemented group were fed the same basal diet as that of the control group, additionally they were supplemented with flaxseed oil @ 5 g/100 g concentrate mixture. A total of 45 semen samples (5 collections from each ram), were collected through electro-ejaculator method (twice in a week and in 3 days interval). The volume, initial motility, mass motility, live and dead percentage, concentration, acrosome intactness and functional membrane integrity were assessed in fresh semen. Sperm velocity and motility parameters were assessed using CASA. Antioxidant enzyme activities, LPO and seminal plasma protein profiling were estimated in seminal plasma. The results indicated that rams fed with oils (treatment groups) produced semen with higher volume, sperm concentrations, with improved membrane and acrosome integrities. The antioxidant enzyme activities were increased and the oxidative stress reduced as indicated by lowered MDA level in the treatment groups. However, the motility and velocity of spermatozoa did not improve although testosterone increased. Testicular biometric measurements did not differ between control and treatment groups. Marked changes were observed in the seminal plasma protein profile due to feeding of oils. The MALDI-TOF/TOF analysis revealed that the differentially expressed protein spots in the coconut oil group were Interleukin-2, Endothelin-1, Secreted phosphoprotein 24, Caveoilin-I, Interferon gamma and in flaxseed oil group were SRY protein. This study indicated that supplementation of flaxseed and coconut oil at 5% level to the rams improved the protective role against oxidative stress and induced over expression of certain seminal plasma proteins depending upon the oil fed.

Mineral supplementation has greater impact on male reproduction; however, the mechanism of action has not been studied in detail. The present study was aimed to deal with the lacuna in mechanism of action of mineral supplementation on improvement in sperm characteristics. A group of 40 bucks (aged 5 months) were assigned to 10 groups (4 in each group) based on their body weight and fed with concentrate mixture: basal roughage (minimal diet) in equal proportion to all the bucks. Among the 10 groups, one was considered as control, without any additional mineral supplementation, and the remaining 9 were treatment groups (3 groups each in Zn, Cu, and Zn + Cu). In treatment groups, organic Zn was fed in three different doses as 20, 40, and 60 mg/kg DM; organic Cu was fed in three different doses as 12.5, 25, and 37.5 mg/kg DM; and organic Zn + Cu was combinedly supplied as 20 + 12.5, 40 + 25, and 60 + 37.5 based on their mg/kg DM for 8 months period. The neat semen samples were processed for spermatozoal gene (stress- NOS3, HSP70, HIF1A; fertility- MTF1, MTA1, TIMP2, TNFa, and EGFR) expression studies through qRT-PCR and protein profile changes through single- and two-dimensional gel electrophoresis. Significantly, the stress-responsive genes were downregulated, and fertility-related genes were upregulated in treatment groups. A significant correlation had been noticed among the genes studied: HIF1A with MTA1 (P < 0.05) and MTF1 with EGFR, TIMP2, TNFa, and NOS3 (P < 0.01) respectively. The organic Zn and Cu feeding modulated the expression of stress- and fertility-related genes and protein abundance, thereby improved the sperm characteristics.

The study was conducted to assess nano zinc (ZnN) as a feed supplement with an aim to compare the supplemental dose of inorganic zinc (ZnI). ZnN was synthesized from 0.45 molar (M) zinc nitrate [Zn(NO3)2.6H2O] and 0.9 M sodium hydroxide (NaOH) and was confirmed to be of ZnN by TEM-EDAX measurements. Wister albino rats (rats; 84, 53.6 ± 0.65 g) were divided into seven groups (4 replicate with 3 rats each) and given feed supplemented with zinc for 60 days with either of the following diets: (1) normal control (NC): basal diet (BD) + no supplemental Zn; (2) ZnI-25: BD + 25 mg/kg Zn from inorganic ZnO; (3) ZnN-25: BD + 25 mg/kg of ZnN; (4) ZnN-12.5: BD + 12.5 mg/kg of ZnN; (5) ZnN-6.25: BD + 6.25 mg/kg of ZnN; (6) ZnN-3.125: BD + 3.125 mg/kg of ZnN; (7) ZnN-50: BD + 50 mg/kg of ZnN. T3 and insulin-like growth factor-1 (IGF-1) hormone levels were similar among groups (P > 0.05), whereas T4 and testosterone were significantly affected, based on supplemented dose. Zn supplementation improved both cell-mediated and humoral immunity. However, both cell-mediated immunity at 24 h and humoral immunity were statistically similar in ZnI-25 and ZnN-6.25 groups. Superoxide dismutase 1 gene expression was found to be similar in all experimental groups. The vascular degeneration were found in liver tissues moderately in NC, mildly in ZnN-6.25 and ZnN-3.125 groups, and no observable changes were noticed in kidney and spleen tissues. However, there was a mild damage in intestinal epithelium of ZnN-25 group rats, hyperplasia of goblet cells, and moderate damage in intestinal villi were observed in ZnN-50 group rats. From the study, it can be concluded that ZnN at half the dose of ZnI showed similar or better responses in terms of immunity, SOD-1 expression, hormonal profiles, and the tissue architecture of vital organs in rats, i.e., 25 mg/kg of Zn from ZnI and 12.5 mg/kg of ZnN impacted similar biological responses like immunity, SOD-1 expression, hormonal profiles, and the tissue architecture of vital organs in rats.

Sex pre-selection of the progeny to suit the livestock production system aids in greatly increasing the efficiency and profitability of the livestock enterprise to meet the demand for animal meat and dairy products. Individual animals can skew the sex ratio in response to various social and environmental stipulations. These social and environmental factors include the composition of the maternal diet, maternal hormonal levels, maternal stress, and the time of insemination. Hence, this present review focuses on these approaches and their success rate as premises for the alterations of offspring sex ratio in different animal species. Numerous studies have linked maternal nutrition and its influence on offspring sex ratio. A maternal diet rich in calcium and magnesium favoured the production of more females, while diets rich in sodium and potassium favoured more males. Furthermore, feeding animals with a particular type of fat diet, including omega-3, omega-6, or omega-9 fatty acids, assists in sex pre-selection. The circulating hormone level in females during conception is associated with the skewing of sex ratio. In addition, maternal stress during the conception period is also found to be associated with the skewing of the sex ratio in favour of female offspring. The timing of insemination relative to the ovulation period has been identified as a promising approach for the pre-selection of offspring sex; however, proper detection of oestrus is crucial. Among all the approaches reported, dietary manipulation could be regarded as the most affordable and stands a better chance of being optimised for commercial livestock production. Reduction in the cost of desired sex production by adopting sex pre-selection techniques will lead to the improvement of the economy and minimise the culling of the undesired sex. However, further research is necessary to focus on the understanding of the underlying molecular mechanisms for the production of animals of the desired sex.

The great expectation of the dairy owners from the frozen semen industry is quality semen with high fertility. Many researchers concentrate in this area to produce frozen semen of high quality and fertility. However, the problem still exists to minimize or reduce the cryoinjury related damages that occur during the process of cryopreservation with the use of all kinds of semen additives. The success rate of the additives added semen is variable at a larger scale. Therefore, any new molecules which can effectively prevent or minimize the cryoinjury are warranted and it could be low cost substances. Our incidental finding and protein identification with molecular weight ≈ 19 kDa (cold-inducible RNA-binding protein CIRBP) in goat spermatozoa seems to be effective in minimizing cryoinjury. The identified CIRBP was confirmed via Western blotting. In future, further studies are needed to confirm the molecular mechanism of the CIRBP on the cryoinjury as semen additives, and application of this protein added semen in the field to improve the level of conception rate in cattle.