Enhancing the quality of inferior oocytes of buffalo for in vitro embryo production: The impact of melatonin on maturation, SCNT, and epigenetic modifications.

Success of animal cloning is limited by oocyte quality, which is closely linked to reprogramming ability. The number of layers of cumulus cells is typically used to assess the quality of oocyte; a minimum of one-third of collected cumulus-oocyte complexes (COCs) are discarded as inferior oocytes because they have less cumulus cells. Melatonin, which has been recognised for its ability to sequester free radicals and perform multiple functions, has emerged as a potentially effective candidate for enhancing inferior oocytes quality and, consequently, embryo development competency. The current study investigates to improve the quality of inferior oocytes by supplementation of melatonin (10-9 M) during in vitro maturation (IVM) and subsequent cloned embryo production and its mechanism. The results indicate that melatonin supplementation significantly (p<0.05) enhances inferior oocytes maturation, reduces oxidative stress by reducing ROS levels, and improves mitochondrial function by boosting GSH levels. The melatonin treatment (10-9 M) enhances the expression of SOD, GPx1, GDF 9, BMP 15, ATPase 6, and ATPase 8 in inferior oocytes. Furthermore, melatonin treatment increases the total cell number in the treated groups, promoting cloned blastocyst formation rates derived from inferior oocytes. Furthermore, compared to the control, 10-9 M melatonin supplementation enhances H3K9ac acetylation and lowers H3K27me3 methylation in cloned blastocysts derived from inferior oocytes. In conclusion, 10-9 M melatonin supplementation during IVM increased inferior oocyte maturation and promoted cloned buffalo embryo development by lowering oxidative stress and promoting epigenetic alterations. These studies show that melatonin may improve the quality of poor oocytes and buffalo cloning.

Partial purification of bacterial cellulo-xylanolytic enzymes and their application in deinking of photocopier waste paper.

The potential of alkaline cellulo-xylanolytic enzymes from non-pathogenic Bacillus subtilis strain was tested for deinking of photocopier waste paper. Cellulase and xylanase play a crucial role in deinking of different types of waste paper. Partial purification of cellulo-xylanolytic enzymes was carried out using ultrafiltration followed by ammonium sulfate precipitation. The ultrafiltered enzyme was used for deinking the photocopier waste paper along with chemical deinking. An enzyme dose of 0.6 IU/g and reaction time of 60 min for ultrafiltered cellulo-xylanolytic enzyme significantly increased deinking efficiency, tear index (9.52%) and folding endurance (5±2%) as compared to chemical deinking. There was improvement in strength properties such as tear index and double-fold along with freeness of pulp (18%). There was slight decrease in tensile index (0.6%) and burst index (16%) while ISO brightness remained unaffected. Enzymatic deinking (74.3%) by ultrafiltered cellulo-xylanolytic from Bacillus subtilis was found significant over conventional chemical deinking.

Bacterial cellulase treatment for enhancing reactivity of pre-hydrolysed kraft dissolving pulp for viscose.

To improve the process economy of reactivity improvement, crude cellulase from Bacillus subtilis was employed for the treatment and significant dissolving pulp properties were analyzed. With increase in enzyme dose from 0.25 to 2 U/g o.d. pulp, improvement in Fock reactivity and alkali solubilities (S10 and S18) were observed with simultaneous reduction in viscosity and yield. Fourier transform infrared spectroscopy and scanning electron microscopy were used to observe the molecular level effects on dissolving grade pulp. The most suitable cellulase dose for reactivity improvement with lowering of viscosity was 0.25 U/g o.d. pulp. With increases in enzyme dose, alkali solubilities (S10 and S18) of dissolving pulp showed continuous increment, while alpha-cellulose of pulp showed reduction due to chain scission of long cellulose fiber fraction.