Prediction of Nili-Ravi buffalo bull fertility through Fourier harmonic analysis of sperm.

Fourier harmonic analysis (FHA) is a robust method for identification of minute changes in sperm nuclear shape that are indicative of reduced fertility. The current study was designed to develop a fertility prediction model for Nili-Ravi buffalo bulls through FHA of sperm. In experiment I, FHA technique was standardized, average sperm nuclear perimeter was measured and sperm nuclear shape plot of buffalo bull was constructed. Sperm of buffalo bulls (n = 10) were stained with YOYO-1 and Hoechst-33342 to differentiate live and dead, and digital images were captured using phase contrast and fluorescent microscopy. The images were analyzed by ImageJ software and 100 sperm/bull were evaluated. The results are described as mean ± SEM values of mean harmonic amplitude (mharm), skewness harmonic amplitude (skharm), kurtosis harmonic amplitude (kurharm) and variance harmonic amplitude (varharm) at Fourier frequencies 0-5 along with the cartesian and polar coordinate plots of buffalo bull sperm. In experiment II, a fertility prediction model was developed based on FHA of buffalo bull sperm. Semen samples of low (n = 6), medium (n = 3) and high (n = 8) fertility bulls were investigated for FHA of sperm and harmonic amplitudes (HA) were generated. Firstly, to determine if live and dead sperm population have unique nuclear shape distribution; the mean, skewness, kurtosis and variance HA 0-5 of 1700 live and 1294 dead spermatozoa of 17 bulls were evaluated. T-test signified a difference in the mharm0 (2.363 ± 0.01 vs. 2.439 ± 0.02), skharm0 (-0.0002 ± 0.07 vs. -0.266 ± 0.09), kurharm0 (-0.156 ± 0.07 vs. 0.260 ± 0.18), kurharm2 (0.142 ± 0.11 vs. 1.031 ± 0.32) and varharm4 (0.109 ± 0.00 vs. 0.082 ± 0.00) of live vs. dead sperm population (p < 0.05). Therefore, 100 live sperm/bull were further evaluated for mean, skewness, kurtosis and variance HA 0-5 values among high (n = 6) and low-fertility (n = 6) groups. Results of T-test showed higher values of mharm2 (0.739 ± 0.01 vs. 0.686 ± 0.00), mharm4 (0.105 ± 0.001 vs. 0.007 ± 0.001), and skharm0 (0.214 ± 0.109 vs. -0.244 ± 0.097) in high vs. low-fertility group (p < 0.05). In next step, five significantly different combinations of discriminant measures between high and low-fertility groups were obtained by discriminant analysis. In conclusion, mharm4, skharm0 and varharm2 correctly identified 91.7 % of bulls into their respective fertility groups, and upon cross validation the value of the canonical correlation was 0.928.

Evaluation of carboxylated poly-l-lysine for cryopreservation of Labeo rohita sperm.

Carboxylated poly-l-lysine (CPLL) is an anti-freeze agent having pronounced non-permeating yet membrane stabilizing cryoprotective capabilities. The objective was to evaluate the CPLL supplementation in extender in terms of post-thaw quality (sperm), total anti-oxidant activity (milt) and fertilization potential of cryopreserved Labeo rohita sperm. For this purpose, male brood fish reared at a fish seed hatchery, Rawal Town Islamabad, Pakistan were captured from different rearing ponds and acclimatized in hatchery ponds for 6 h. The brooder was injected with Ovaprim (0.2 mL/kg), and milt was collected after 8 h in cooled sterilized falcon tubes, maintained at 4°C and evaluated for sperm motility. The milt collected from three brooders (n = 3) was diluted in extenders viz., modified Kurokura-2 extender having 10% methanol (control); experimental extenders with CPLL supplementation at the rate of 0.5%, 1% and 1.5%. Diluted milt was filled in 0.5 mL straws, exposed to liquid nitrogen vapours and cryopreserved. Cryopreserved milt was thawed at 25°C and assessed for post-thaw sperm quality. Sperm motility, motility duration, viability, total anti-oxidant capacity and DNA integrity was significantly higher (p < 0.05) in the extender having 1.5% CPLL than control. To evaluate the fertilization rates, male and female brooders were injected with Ovaprim at 0.2 mL/Kg and 0.5 mL/Kg body weight respectively. Fresh eggs and milt were collected through abdominal stripping. Batches of 10 g of eggs from each female (n = 2) were fertilized with one straw, each from frozen sperm with KE + methanol (control), KE + methanol + 1.5% CPLL and 50 µL fresh milt (negative control). After 1.5 h of fertilization, eggs were collected from all jars and a total of 200 eggs were counted. The fertilized eggs appeared clear and transparent while unfertilized eggs looked opaque with disintegrated nuclei. Sperm fertilization rate (%) was higher (p < 0.05) in extender KE + methanol + 1.5% CPLL (78.7 ± 0.5) compared to control (KE + methanol) (52.0 ± 0.4) however, it was lower compared to that of negative control, the fresh milt (85.2 ± 0.6). In conclusion, supplementation of carboxylated poly-l-lysine (1.5%) to modified Kurokura-2 extender having 10% methanol improves post-thaw motility, motility duration, viability, DNA integrity, anti-oxidant capacity (milt) and fertilizing ability of cryopreserved L. rohita sperm.

Metal ferrites-based nanocomposites and nanohybrids for photocatalytic water treatment and electrocatalytic water splitting.

Photocatalytic degradation is one of the most promising technologies available for removing a variety of synthetic and organic pollutants from the environmental matrices because of its high catalytic activity, reduced energy consumption, and low total cost. Due to its acceptable bandgap, broad light-harvesting efficiency, significant renewability, and stability, Fe2O3 has emerged as a fascinating material for the degradation of organic contaminants as well as numerous dyes. This study thoroughly reviewed the efficiency of Fe2O3-based nanocomposite and nanomaterials for water remediation. Iron oxide structure and various synthetic methods are briefly discussed. Additionally, the electrocatalytic application of Fe2O3-based nanocomposites, including oxygen evolution reaction, oxygen reduction reaction, hydrogen evolution reaction, and overall water splitting efficiency, was also highlighted to illustrate the great promise of these composites. Finally, the ongoing issues and future prospects are directed to fully reveal the standards of Fe2O3-based catalysts. This review is intended to disseminate knowledge for further research on the possible applications of Fe2O3 as a photocatalyst and electrocatalyst.

Sol-Gel Synthesized High Entropy Metal Oxides as High-Performance Catalysts for Electrochemical Water Oxidation.

Hexanary high-entropy oxides (HEOs) were synthesized through the mechanochemical sol-gel method for electrocatalytic water oxidation reaction (WOR). As-synthesized catalysts were subjected to characterization, including X-ray diffraction (XRD), Fourier transforms infrared (FTIR) analysis, and scanning electron microscopy (SEM). All the oxide systems exhibited sharp diffraction peaks in XRD patterns indicating the defined crystal structure. Strong absorption between 400-700 cm-1 in FTIR indicated the formation of metal-oxide bonds in all HEO systems. WOR was investigated via cyclic voltammetry using HEOs as electrode platforms, 1M KOH as the basic medium, and 1M methanol (CH3OH) as the facilitator. Voltammetric profiles for both equiatomic (EHEOs) and non-equiatomic (NEHEOs) were investigated, and NEHEOs exhibited the maximum current output for WOR. Moreover, methanol addition improved the current profiles, thus leading to the electrode utility in direct methanol fuel cells as a sequential increase in methanol concentration from 1M to 2M enhanced the OER current density from 61.4 to 94.3 mA cm-2 using NEHEO. The NEHEOs comprising a greater percentage of Al, ([Al0.35(Mg, Fe, Cu, Ni, Co)0.65]3O4) displayed high WOR catalytic performance with the maximum diffusion coefficient, D° (10.90 cm2 s-1) and heterogeneous rate constant, k° (7.98 cm s-1) values. These primary findings from the EC processes for WOR provide the foundation for their applications in high-energy devices. Conclusively, HEOs are proven as novel and efficient catalytic platforms for electrochemical water oxidation.

Sperm binding to hyaluronan is an excellent predictor of Nili-Ravi buffalo bull fertility.

This study reports the first evaluation of sperm hyaluronan binding assay (HBA) for predicting the fertility of Nili-Ravi buffalo bulls in relation to standard parameters of sperm quality. Cryopreserved semen doses of low (n = 6), medium (n = 3) and high fertility (n = 8) bulls based on their respective return rates were used. Significantly, more spermatozoa bound to hyaluronan from the most fertile bulls (57.15% ± 1.44) compared with medium (42.46% ± 1.08) and low fertility bulls (29.70% ± 0.78). A strongly positive correlation (r = .824, p < .01) was found between HBA and fertility that predicts a 67.9% variability (r2  = .679, p < .01) in fertility. HBA was also strongly positively correlated with sperm viability (r = .679, p < .01) followed by their live/dead ratio (r = .637, p < .01), uncapacitated spermatozoa (r = .631, p < .01), normal apical ridge (r = .459, p < .01), motility (r = .434, p < .01), mature spermatozoa with low residual histones (r = .364, p < .01), high plasma membrane integrity (r = .316, p < .01) and nonfragmented DNA levels (r = .236, p < .05). It was negatively correlated with spermatozoa having reacted acrosome (r = -.654, p < .01). A fertility model built using a combination of sperm HBA and either sperm livability or viability predicts, respectively, 86.1% (r2  = .861, p < .01) and 85.9% (r2  = .859, p < .01) variability in buffalo bull fertility. In conclusion, sperm HBA may prove to be a single robust predictor of Nili-Ravi buffalo bull fertility.

Effect of Synergism Between Carboxylated Poly-l-Lysine and Glycerol on Freezability of Nili-Ravi Buffalo (Bubalus bubalis) Semen.

Carboxylated poly-l-lysine (CPLL), an ampholytic polymer, has remarkable cryoprotective properties. It was hypothesized that CPLL will reduce/replace glycerol in extender and improve the freezability of buffalo semen. The objective was to evaluate various combinations of CPLL and glycerol in extenders for any synergism toward cryosurvivability of Nili-Ravi buffalo sperm. Semen collected from four Nili-Ravi buffalo bulls (ejaculates = 40; replicates = 5) was diluted in tris-citric acid based extenders, Control: (0% CPLL +7% Glycerol); E1: (1% CPLL +6% Glycerol); E2: (2% CPLL +5% Glycerol); E3: (3% CPLL +4% Glycerol); E4: (4% CPLL +3% Glycerol); E5: (5% CPLL +2% Glycerol); E6: (6% CPLL +1% Glycerol), and E7: (7% CPLL +0% Glycerol), and cryopreserved using a programmable cell freezer. Percentages of post-thaw sperm motility, plasma membrane integrity, and acrosomal integrity were found to be higher (p < 0.05) in extenders E1, E2, E3, E4, and E5 compared to E6 and the control. Sperm livability (%; live/dead ratio) and viability (%; live sperm with intact acrosome) were higher (p < 0.05) in extender E4 compared to all the other extenders. Sperm DNA integrity was higher (p < 0.05) in extender E2, E3, E4, and E5 compared to control, E1, and E6 extenders. Sperm lipid peroxidation levels were lower (p < 0.05) in E3, E4, E5, and E6 compared to control, E1, E2, and E7 extenders. Total antioxidant capacity of seminal plasma was higher (p < 0.05) in extenders E5, E6, and E7 than control, E1, E2, E3, and E4 extenders. It is concluded that synergism between CPLL and glycerol (4% CPPL and 3% Glycerol) seems to improve the freezability of Nili-Ravi buffalo semen by reducing oxidative stress.