The utility of nanowater for ram semen cryopreservation

Abstract

Nanowater (NW; water declusterized in the low-temperature plasma reactor) has specific physicochemical properties that could increase semen viability after freezing and hence fertility after artificial insemination (AI) procedures. The main goal of this study was to evaluate ram semen quality after freezing in the media containing NW. Ejaculates from 10 rams were divided into two equal parts, diluted in a commercially available semen extender (Triladyl Õ ; MiniTü b GmbH, Tiefenbach, Germany) prepared with deionized water (DW) or NW, and then frozen in liquid nitrogen. Semen samples were examined for sperm motility and morphology using the sperm class analyzer system and light microscopy. Cryo-scanning electron microscopy (cryo-SEM) was employed to determine the size of extracellular water crystals in frozen semen samples. Survival time at room temperature, aspartate aminotransferase (AspAT) and alkaline phosphatase (ALP) concentrations post-thawing as well as conception/lambing rates after laparoscopic intrauterine AI of 120 ewes were also determined. There were no significant differences between DW and NW groups in sperm progressive motility (26.4 AE 12.2 and 30.8 AE 12.4%) or survival time (266.6 AE 61.3 and 270.9 AE 76.7 min) after thawing and no differences in the percentages of spermatozoa with various morphological defects before or after freezing. There were, however, differences (P < 0.05) in AspAT (DW: 187.1 AE 160.4 vs. NW: 152.7 AE 118.3 U/l) and ALP concentrations (DW: 2198.3 AE 1810.5 vs. NW: 1612.1 AE 1144.8 U/l) in semen samples post-thawing. Extracellular water crystals were larger (P < 0.05) in ejaculates frozen in NW-containing media. Ultrasonographic examinations on day 40 post-AI revealed higher (P < 0.05) conception rates in ewes inseminated with NW (78.3%) compared with DW semen (58.3%), and the percentages of ewes that carried lambs to term were 73.3% and 45.0% in NW and DW groups, respectively (P < 0.01). In summary, the use of a semen extender prepared with NW was associated with a substantial improvement in the fertilizing ability of frozen-thawed ram semen and lamb productivity of inseminated ewes.

The Water Treated With Low-Frequency Low-Pressure Glow Plasma Enhances the Phytoavailability of Selenium and Promotes the Growth of Selenium-Treated Cucumber Plants

“…and d.w. of the shoots and roots were significantly higher in the DWbased than in the GPTW-based medium. The cause of this phenomenon requires further clarification, but we suppose that it may be associated with better solubility of mineral salts in GPTW than in DW [Murawski et al 2015], which can modify the bioavailability of essential mineral nutrients from the solution. However, when The mean values (n = 6 for the concentration of photosynthetic pigments, and n = 12 for Fv/Fm value) in each column followed by different letters indicate significant differences between treatments (Tukey's test; p ˂ 0.05).…”

“…2). The enhanced Se accumulation in the GPTW-based medium, especially in the roots, may result from the higher Se solubility in GPTW than in DW [Murawski et al 2015] or/and from the presence inside the GPTW clathrates of excited, probably singlet oxygen molecules [Bialopiotrowicz et al 2016], which can modify the oxidation state and/or balance of Se ions in the medium, thus influencing the availability and translocation of Se within plants. It is known that Se in the form of selenite (Se 4+ ) is rapidly converted to organoselenium compounds that are accumulated in the root system, whereas selenate (Se 6+ ) is delivered immediately to the xylem and assimilated into organoselenium compounds in plastids [White 2015].…”

“…Moreover, GPTW has low viscosity, high diffusivity, and very low density. It dissolves 35-40% more substances than the same volume of normal water, thus it can be a very efficient carrier of mineral nutrients [Mystkowska et al 2013, Murawski et al 2015.…”

Current status and advances in ram semen cryopreservation

“…Arav et al (2002) developed a device (Multi-Thermal-Gradient'-MTG ® ) to control temperature seeding and morphology of created ice crystals, achieving high rates of post thaw semen motility. Recently, Murawski et al (2015) used an innovative technology preparing the commercial semen extender Triladyl ® (Minitube, Germany) with water declusterized in the low-temperature plasma reactor, called "nanowater". This process led not only to lower crystallization rate of water but also to a substantial improvement in the fertilizing ability of frozen-thawed ram semen and higher conception rates after laparoscopic AI of ewes.…”

Pool Boiling Heat Transfer Coefficient of Low-Pressure Glow Plasma Treated Water at Atmospheric and Reduced Pressure

“…The possibilities of using GPTW are shown in Figure 1. The use of GPTW has already been reported in relation to disinfection [24] and the cryopreservation of biological samples [25]. This type of water is also used in cosmetology, due to the possibility of dissolving more chemical compounds, and even allows the dissolution of compounds that are practically insoluble in plain water, which makes it possible to obtain highly-concentrated aqueous solutions of active substances.…”