Improvement of diluted semen from Muscovy (Cairina moschata) drakes by the addition of water-soluble antioxidants.

The aim of this study was to evaluate the effect of antioxidant supplementation in diluted semen from Muscovy drakes after the induction of oxidative stress (OS) on the sperm motility, kinematic parameters and biochemical markers - lipid peroxidation (LPO) levels and total glutathione (tGSH) concentration. The pooled semen was distributed equally into three parts, diluted (1:3 v/v) with IMV Canadyl, HIA-1 or AU, and stored at 4°C for 6 h. Later, the semen was equilibrated at 20-25°C for 15 min, and divided in Eppendorf tubes. The sperm samples (final concentration of 50 × 106 sperm cells/mL per sample) were incubated at 37°C for 30 min in the absence (-) or presence (+) of 0.1 mM FeSO4 + 0.5 mM H2 O2 (Fenton system) and the following combinations of antioxidants: ascorbic acid + Trolox (A + T); ascorbic acid + Desferal (A + D); Trolox + Desferal (T + D) and ascorbic acid + Trolox + Desferal (A + T + D), all of them in a final concentration of 0.1 mM. Thus, the total number of samples was 30 and in each one, the sperm motility, velocity parameters, LPO and tGSH were determined. The motility and kinematic parameters of the diluted semen with added antioxidants were restored by up to 20% after inducing OS via the Fenton reaction. Dual combinations of antioxidants (A + T, A + D, and T + D) lowered LPO levels, but not equally across different extenders. After the induction of OS, the tGSH levels in diluted semen with IMV-Canadyl were not affected by the added antioxidants. Whereas antioxidant combinations in diluted semen with HIA-1 or AU had a beneficial effect and partially restored tGSH levels. In conclusion, the results showed that the extender IMV-Canadyl is well balanced and protected the Muscovy semen under OS conditions, while the other two extenders HIA-1 and AU can be improved by adding antioxidants.

Effect of Chitosan-Diosgenin Combination on Wound Healing.

The difficult-to-heal wounds continue to be a problem for modern medicine. Chitosan and diosgenin possess anti-inflammatory and antioxidant effects making them relevant substances for wound treatment. That is why this work aimed to study the effect of the combined application of chitosan and diosgenin on a mouse skin wound model. For the purpose, wounds (6 mm diameter) were made on mice's backs and were treated for 9 days with one of the following: 50% ethanol (control), polyethylene glycol (PEG) in 50% ethanol, chitosan and PEG in 50% ethanol (Chs), diosgenin and PEG in 50% ethanol (Dg) and chitosan, diosgenin and PEG in 50% ethanol (ChsDg). Before the first treatment and on the 3rd, 6th and 9th days, the wounds were photographed and their area was determined. On the 9th day, animals were euthanized and wounds' tissues were excised for histological analysis. In addition, the lipid peroxidation (LPO), protein oxidation (POx) and total glutathione (tGSH) levels were measured. The results showed that ChsDg had the most pronounced overall effect on wound area reduction, followed by Chs and PEG. Moreover, the application of ChsDg maintained high levels of tGSH in wound tissues, compared to other substances. It was shown that all tested substances, except ethanol, reduced POx comparable to intact skin levels. Therefore, the combined application of chitosan and diosgenin is a very promising and effective medication for wound healing.

Changes in the Oxidative Status of Dual-Purpose Hens Rearing in the Free-Range System during Cold, Thermoneutral and Hot Period.

This study aimed to assess the changes in the oxidative status of six genotypes of free-range laying hens during cold, thermoneutral, and hot periods by measuring the levels of lipid peroxidation (LPO), total glutathione (tGSH), and the activity of antioxidant enzymes catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) in erythrocyte suspension, in relation with their egg production. Two identical experiments were conducted in two consecutive years. Thermal stress adversely affected the oxidative status of hens. The induced OS is expressed by an increase in LPO and the activities of antioxidant enzymes SOD and GPx during cold and hot periods and a decrease in CAT and tGSH during the cold period in both years. The factor "temperature period", compared to "year" and "genotype", had the most significant influence on all biochemical parameters (p < 0.001). Significant phenotypic correlations (p < 0.05) were detected among studied biochemical parameters, except between SOD and tGSH. The chicken genotypes showed differences in their susceptibility to OS and this had an effect on egg production­from 37.87% to 74.93%. The OS is genotypically specific and can play a significant role in determining welfare and egg production in free-range systems.

Effect of prooxidants and chelator Desferal on the oxidative status and sperm motility of Muscovy semen.

This study aimed to establish the sensitivity of Muscovy duck semen to oxidative stress (OS) and the effect of Desferal, applied as an antioxidant. The effect of three prooxidant systems in presence and absence of Desferal were tested on the motility and kinetic parameters (determined using CASA system), as well as the level of lipid peroxidation (LPO) and glutathione (tGSH) of Muscovy semen. The semen was diluted (1:3 v/v) with four extenders (saline solution, IMV Canadyl, HIA-1, and AU) and stored at 4 °C for 6 h. The cooled semen was divided into aliquots (50 × 106 sperm cells/mL), which were incubated at 37 °C for 30 min with one of the following prooxidative agents: ferrous sulfate (FeSO4, 0.1 mM), hydrogen peroxide (H2O2, 1 mM), and Fenton system (FeSO4(Fe2+), 0.1 mM + H2O2, 1 mM), in the presence or absence of Desferal (0.1 mM). The addition of FeSO4 + H2O2 or FeSO4 regardless of the used extender, as well as the addition of H2O2 to the diluted semen with saline solution significantly increased the levels of LPO (P < 0.05). With the lowest prooxidant effect was H2O2. The application of Desferal reduced significantly (P < 0.05) the LPO levels induced by FeSO4 + H2O2 or FeSO4 and in a weaker degree by H2O2. Among all prooxidants, FeSO4 + H2O2 decreased in the greatest extent the tGSH concentration in semen diluted with each used extenders in comparison to the relevant control. The addition of Desferal in semen diluted with HIA-1 extender and incubated with FeSO4, and H2O2, showed the best restoration of tGSH level, close to that of respectively controls. The studied prooxidants significantly reduced total, progressive, and kinetic sperm motility (P < 0.05). Although the inclusion of Desferal reduced the sperm OS, it did not improve the impaired by OS sperm motility.