Effects of five cryoprotectants on proliferation and differentiation-related gene expression of frozen-thawed bovine calf testicular tissue.

The cryopreservation of testicular tissue is a potential method for preserving male fertility. However, the effect of cryopreservation on bovine calf testicular tissue is scarce. This study investigated the effect of different cryoprotectants on bovine calf testicular tissue at the molecular level. Testicular tissue from ten immature bovine calves (6 months) was collected after slaughter and cryopreserved in an extender containing different concentrations of the following five cryopreservation solutions (CP): bovine serum albumin (BSA) with 5% dimethyl sulfoxide (DMSO), trehalose with 5% DMSO, DMSO and glycerol and ethylene glycol (EG). After 7-day cryopreservation, the expression levels of three spermatogonial stem cell (SSC)-related genes, octamer-4 (OCT4), KIT ligand (MGF/SCF) and kit oncogene (C-KIT), were investigated by quantitative PCR (qPCR). The cell viability was highest for the tissues preserved with 30 mg/ml BSA (77.82% ± 1.22) and 40 mg/ml trehalose (74.23% ± 1.16) compared with other groups (p < 0.05), and the level of expression of the three genes was highest with 30 mg/ml BSA (p < 0.05). Compared with other CPs, the 30 mg/ml BSA and 40 mg/ml trehalose have the better cryopreserve protection. The 30 mg/ml BSA is the most viable media for the cryopreservation of testicular tissue from cattle.

Ensiling corn stover: effect of feedstock preservation on particleboard performance.

Ensilage is a truncated solid-state fermentation in which anaerobically produced organic acids accumulate to reduce pH and limit microbial activity. Ensilage can be used to both preserve and pretreat biomass feedstock for further downstream conversion into chemicals, fuels, and/or fiber products. This study examined the ensilage of enzyme-treated corn stover as a feedstock for particleboard manufacturing. Corn stover at three different particle size ranges (<100, <10, and <5 mm) was ensiled with and without a commercial enzyme mixture having a cellulase:hemicellulase ratio of 2.54:1, applied at a hemicellulase rate of 1670 IU/kg dry mass. Triplicate 20 L mini-silos were destructively sampled and analyzed on days 0, 1, 7, 21, 63, and 189. Analysis included produced organic acids and water-soluble carbohydrates, fiber fractions, pH, and microorganisms, including Lactobacillus spp. and clostridia were monitored. On days 0, 21, and 189, the triplicate samples were mixed evenly and assembled into particleboard using 10% ISU 2 resin, a soy-based adhesive. Particleboard panels were subjected to industry standard tests for modulus of rupture (MOR), modulus of elasticity (MOE), internal bonding strength (IB), thickness swell (TS), and water absorption at 2 h boiling and 24 h soaking. Enzyme addition did improve the ensilage process, as indicated by sustained lower pH (P < 0.0001), higher water-soluble carbohydrates (P < 0.05), and increased lactic acid production (P < 0.0001). The middle particle size range (<10 mm) demonstrated the most promising results during the ensilage process. Compared with fresh stover, the ensilage process did increase IB of stover particleboard by 33% (P < 0.05) and decrease water adsorption at 2 h boiling and 24 h soaking significantly (P < 0.05). Particleboard panels produced from substrate ensiled with enzymes showed a significant reduction in water adsorption of 12% at 2 h boiling testing. On the basis of these results, ensilage can be used as a long-term feedstock preservation method for particleboard production from corn stover. Enzyme-amended ensilage not only improved stover preservation but also enhanced the properties of particleboard products.