Summary Information on donkey sperm bioenergetics, kinetics and oxidative status is scarce even though crucial for development of reproductive technologies and germplasm conservation. For these reasons, it is interesting to monitor sperm kinetics, bioenergetics, and oxidative status during sperm storage at +4°C and with several sperm extenders and concentrations. Donkey semen was collected from three jackasses, three times each. It was diluted with four extenders (Kenney, Equiplus, INRA96 or Hippex), set at three sperm concentrations (30, 50 or 70 × 106 spermatozoa/ml) and evaluated for its functionality after 0, 3, 24, 48 and 72 h storage at +4°C. Sperm kinetics was analyzed by Sperm Computer Analysis; sperm bioenergetics was assessed by mitochondrial membrane potential (MMP); sperm oxidative status was evaluated by lipid peroxidation (LPO), anti-LPO potential and nitroblue tetrazolium (NBT) assays. Incubation produced a progressive (P < 0.01) decline in sperm kinetics and MMP, whereas parameters related to oxidative status either increased (LPO, NBT) or decreased (anti-LPO). The anti-LPO potential was the index better related to sperm motility and kinetics. Extenders proved to be differently (P < 0.01) effective in preserving sperm kinetics, MMP, and oxidative status. The concentration of 30 × 106 spermatozoa/ml provided an optimum preservation of sperm functions. Significant correlations emerged between most parameters examined. This study identified reference criteria for storing donkey spermatozoa at +4°C. A low sperm concentration together with a proper extender are crucial requirements for optimum sperm cryopreservation efficiency. Field trials are, however, required to validate these findings, making them operational in practice.
By exploiting the PMN property to produce high quantities of oxygen peroxide to neutralize pathogens, the oxygen peroxide content of uterine cells was measured to diagnose endometritis. After preliminary in vitro studies in which endometrial cells from slaughtered mares were mixed with leukocytes from peripheral blood, endometrial samples were collected by uterine flushing from mares before insemination. Staining endometrial cells with H2DCF-DA was combined with hydroethidine to normalize the fluorescence intensity with the cellular content of the sample. Stained cell smears were assumed as the gold standard of endometritis, and based on this assay, the samples were considered positive (C+) and negative (C−) for endometritis. The amount and the turbidity of fluid recovered by uterine flushing were significantly (p < 0.01) higher in C+ than in C−. Moreover, the oxygen peroxide content of the endometrial cells was significantly higher in the C+ than in the C− group (6.31 ± 1.92 vs. 3.12 ± 1.26, p = 0.001). Using the value of 4.4 as the cutoff level of this fluorescence cytology assay, it was found that only one C− sample exceeded the cutoff level (false positives = 7.7%) while three C+ samples showed values below the cutoff level (false negative = 11.5%).
Zn-bearing phyllosilicates are common minerals in nonsulfide Zn deposits, but they seldom represent the prevailing economic species. However, even though the presence of Zn-bearing clays is considered as a disadvantage in mineral processing, their characteristics can give crucial information on the genesis of the oxidized mineralization. This research has been carried out on the Mina Grande and Cristal Zn-sulfide/nonsulfide deposits, which occur in the Bongará district (Northern Peru). In both of the deposits, Zn-bearing micas and clays occur as an accessory to the ore minerals. The XRD analyses and scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDS) investigations revealed that the Zn-bearing micas that are occurring in both deposits mostly consist of I/S mixed layers of detrital origin, which have been partly altered or overprinted by sauconite during the supergene alteration of sulfides. Sporadic hendricksite was also identified in the Cristal nonsulfide mineral assemblage, whereas at Mina Grande, the fraipontite-zaccagnaite (3R-polytype) association was detected. The identified zaccagnaite polytype suggests that both fraipontite and zaccagnaite are genetically related to weathering processes. The hendricksite detected at Cristal is a product of hydrothermal alteration, which is formed during the emplacement of sulfides. The complex nature of the identified phyllosilicates may be considered as evidence of the multiple processes (hydrothermal and supergene) that occurred in the Bongará district.
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