Glycerol Treatments Research Articles

Tunable Fungal Monofilaments from Food Waste for Textile Applications.

A fungal biorefinery is presented to valorize food waste to fungal monofilaments with tunable properties for different textile applications. Rhizopus delemar is successfully grown on bread waste and the fibrous cell wall is isolated. A spinnable hydrogel is produced from cell wall by protonation of amino groups of chitosan followed by homogenization and concentration. Fungal hydrogel is wet spun to form fungal monofilaments which underwent post-treatments to tune the properties. The highest tensile strength of untreated monofilaments is 65MPa (and 4% elongation at break). The overall highest tensile strength of 140.9MPa, is achieved by water post-treatment. Moreover, post-treatment with 3% glycerol resulted in the highest elongation % at break, i.e., 14%. The uniformity of the monofilaments also increased after the post-treatments. The obtained monofilaments are compared with commercial fibers using Ashby's plots and potential applications are discussed. The wet spun monofilaments are located in the category of natural fibers in Ashby's plots. After water and glycerol treatments, the properties shifted toward metals and elastomers, respectively. The compatibility of the monofilaments with human skin cells is supported by a biocompatibility assay. These findings demonstrate fungal monofilaments with tunable properties fitting a wide range of sustainable textiles applications.

Utilization of Duck Feet Gelatin with the Additional Glycerol as A Plasticizer on the Physical Properties of Edible Film

The packaging that is widely used is plastic packaging, but this material has a weakness that is difficult to decompose and isn’t biodegradable, therefore edible film can be an alternative primary packaging for food products. The material used consists of gelatine made from duck feet with the addition of various concentrations of glycerol. Duck feet is an underutilized portion of the animal's anatomy, yet they contain collagen, which has the potential to be the raw material for gelatine and edible films. The study was undertaken utilizing duck feet as a source of raw material, which was subsequently processed into gelatine for use in the production of edible films using different glycerol treatments. The treatments comprised of glycerol concentrations of P1 = 10%, P2 = 20%, P3 = 30%, P4 = 40%, P5 = 50%, and P6 = 60%. The findings indicated that the application of glycerol significantly affected the films' solubility, water absorption, and tensile strength (P<0.05), but didn’t significantly affect the elongation of edible films. (P>0.05). The optimal concentration of glycerol for edible film in this study was obtained by using 20% glycerol, with a physical characteristic value of 40.25% solubility, 24.23% water absorption, tensile strength is 0.57 MPa, and elongation is 90%.

Fungal textile alternatives from bread waste with leather-like properties

Food waste and fashion pollution are two of the most prominent global environmental issues. To alleviate the problems associated with food waste, while simultaneously contributing to sustainable fashion, the feasibility of making an alternative textile material with leather-like properties from fungal biomass cultivated on bread waste was investigated. The filamentous fungus, Rhizopus delemar, was successfully grown on waste bread in a submerged cultivation process, and fungal biomass was treated with vegetable tannin of chestnut wood. NMR and FTIR confirmed interactions between tannin and fungal biomass, while OM, SEM and AFM visualised the changes in the hyphae upon the tannin treatment. Thermal stability was assessed using TGA analysis. The wet-laid technique commonly utilised for paper-making was used to prepare sheets of hyphae. Some of the sheets were treated with glycerol and/or a biobased binder as post-treatment. Overall, three of the produced materials exhibited leather-like properties comparable to that of natural leather. Sheets from untreated biomass with only glycerol post-treatment showed a tensile strength of 7.7 MPa and an elongation at break of 5%. Whereas sheets from untreated biomass and tannin treated biomass with both glycerol and binder treatments led to tensile strengths of 7.1 MPa and 6.9 MPa, and the elongation at break of 12% and 17%, respectively. The enhancement of hydrophobicity after the binder treatment, helped to preserve the absorbed glycerol within the sheet and thereby the flexibility was retained when in contact with moisture. These findings demonstrate that bread waste-derived fungal sheets have great potential as environmentally friendly materials with leather-like properties.

Increased Comfort of Polyester Fabrics.

The hydrophilicity of fibers is directly related to the comfort of a fabric and represents one of the most important aspects of a textile. Therefore, polyester (PES) modification has focused on an increase in moisture content and a subsequent improvement of the user’s experience. Based on the glycerol hygroscopic properties, the main objective has been the enhancement of the hydrophilicity of polyester by glycerol treatments. Furthermore, microwave irradiation and alkaline treatment have been applied, in order to increase glycerol adhesion. Treated PES samples were characterized by performing moisture content, negative ion, water diffusion and water vapor resistance analyses. The effect of different treatment conditions such as bath ratio (1/10 or 1/15), temperature (40, 60 or 100 °C), time (2 or 5 min) and microwave radiation intensity (300 or 500 W) was evaluated. The moisture content of treated PES results indicated that by decreasing the bath ratio and increasing the time and temperature the moisture gain can reach almost 14%, which can be easily related to increases in the weight of the fiber. The treatment with alkali was done and led to the highest moisture increase. Treatment with 500 W microwave irradiation led to higher glycerol retention after rinsing. Different experimental conditions were applied to the glycerol-treated PES fabrics, and a clear improvement in moisture content was obtained increasing the comfort. The results were compared with the ones obtained for cotton and wool, where the moisture is higher than non treated PES.

Preparation and Performance of Cement Mortar Reinforced by Modified Bamboo Fibers.

This study aims to prepare bamboo-fiber-reinforced cement composites and provide a solution to the issue of poor interfacial adhesion between bamboo fibers and cement matrix. The original bamboo fibers were modified by three moderately low-cost and easy-to-handle treatments including glycerol, aluminate ester, and silane treatments. The performance of the modified bamboo-fiber-reinforced cement composites was evaluated by a series of mechanical and durability tests, including flexural and compressive strength, water absorption, chloride ion penetration, drying shrinkage, freeze–thaw resistance, and carbonization. In addition, the microstructures of composites were characterized using a scanning electron microscope (SEM). The results showed that the composites reinforced with glycerol-modified bamboo fibers had 14% increased flexural strength and comparable compressive strength. From durability perspectives, all treatments showed similar performance in drying shrinkage, whereas aluminate ester treatment was the most effective in terms of impermeability, chloride resistance, freeze–thaw resistance, and carbonization. The results could provide insights to efficient and effective natural fiber treatment to enable better performance of natural-fiber-reinforced cement-based materials.

Urine metabolome alterations in malnutrition and the impact of glycerol or rumen-protected choline chloride supplementation in advanced pregnant ewes

The objective of this study was to explore the metabolic profiles of pregnancy malnutrition induced by feed restriction (FR) and the counteracting effects of glycerol and rumen-protected choline chloride supplementation. Two feeding trials were conducted. In the first experiment, twenty pregnant Hu sheep carrying multiple fetuses with a gestation period of 108 d were randomly divided into two groups. The ewes in the control (CON) group were offered 100 % of their nutritional requirements as recommended by the National Research Council (NRC), while the FR group was offered 30 % of feed intake of CON for 15 d. In the second experiment, eighteen pregnant Hu sheep were offered a feed intake comprising 30 % of the NRC-recommended nutritional requirements twice daily. The sheep were randomly divided into three groups: the FR group in the second experiment (FR2), with no supplementation, the glycerol (GLY) group, which received 40 ml of glycerol per d, and the rumen-protected choline chloride (RPC) group, which received 10 g of rumen-protected choline chloride per d for 9 d. In the first experiment, the urine metabolome of sixteen ewes showed significant difference between the CON group and FR group. Compared with the CON group, FR decreased the level of d-glucose, lactic acid, levoglucosan, α-ketoglutarate, phosphohydroxypyruvic acid, glucose 6-phosphate and the methyl donors, while increasing the level of pyruvate, fumaric acid and carnitines in urine. Both the GLY and RPC treatments counteracted some of these changes and modulated the urine metabolome in advanced pregnant ewes suffering from malnutrition.

Application of Pectin Extracted from Cocoa Pod in the Production of Edible Film

Cocoa pod is lignocellulosic waste which contain cellulose, hemicellulose, lignin, pectin and other compounds. Pectin is a polymer of D-galactaconic acid linked by an α-1,4 glycoside bond. Some carboxyl groups in pectin compounds undergo esterification with methyl (methylation) to methoxyl groups. This compound is referred to as pectinic acid or pectin. Pectin in cocoa pod can be produced by extraction. The purpose of this research is to know the physical and chemical characteristics of pectin-based edible film from cocoa pod. This research is conducted in two steps. The first stage is the extraction of pectin from cocoa pod, and the second stage is the process of making edible film. Pectin extraction was performed by using 5% citric acid. The extracted pectin will be made into an edible film using glycerol as plastisizer. The glycerol treatments are 0%, 2%, 4%, 6%, 8%. The design used was Completely Randomized Design (CRD) at 5% significant level. The results showed that the addition of glycerol has significant effect on water content, thickness, color and water vapor transmission rate.

PRODUCTION AND CHARACTERIZATION OF BIOSURFACTANTS PRODUCED BY Pseudomonas aeruginosa B031 ISOLATED FROM A HYDROCARBON PHYTOREMEDIATION FIELD

The biosurfactants are used by several industrial sectors such as petroleum, agriculture, food production, chemistry, cosmetics, and pharmaceuticals. Because of their hydrophobic and hydrophilic moieties, they have potency to reduce surface tension, interfacial tension between water-hydrocarbon systems, and low micelle concentration. Their characteristics strongly depend on the producer strain as well as on the medium composition, such as carbon and nitrogen sources. This study was conducted to investigate the influence of different sources of carbon (n-hexadecane, glycerol and glucose) and nitrogen (urea, NH4Cl and NaNO3) for the production of biosurfactants by a new strain of Pseudomonas aeruginosa B031 isolated from a rhizosphere of Paraserianthes falcataria L. Nielsen, a hardwood plant species at a phytoremediation field. The biosurfactant characteristics of the strain were evaluated, particularly its surface-active properties and potential to remove hydrocarbon. Glycerol was found to be the optimum carbon source, with rhamnose concentration, emulsification index, and critical micelle concentration (CMC) of 718 mg/L, 37%, and 35 mN/m, respectively. NaNO3 was observed as the optimum nitrogen source, with rhamnose concentration, emulsification index, and CMC of 290 mg/L, 30%, and 24 mN/m, respectively. These biosurfactants efficiently reduced surface tension of culture broth from 42 mN/m to 31 mN/m for the glycerol treatment and from 37 mN/m to 24 mN/m for the sodium nitrate treatment. The crude biosurfactants from the glycerol and sodium nitrate treatments also removed 87.5% and 84%, respectively, of crude oil from sand. These rates were higher than those of the chemical surfactants (SDS and Triton X-100). These findings indicate that the biosurfactants produced by the strain from both glycerol and NaNO3 treatments can efficiently decrease the interfacial tension of culture broth dilution and have a high emulsion index, thus hold promise in hydrocarbon bioremediation application.

92 Sperm Cryopreservation in the Burmese Python (Python bivittatus) as a Model for Endangered Snakes

The Burmese python (Python bivittatus) is listed as vulnerable by the International Union for Conservation of Nature (IUCN). Released pet Burmese pythons have detrimental effects on fauna native to southern Florida and are responsible for localised declines of several species in some parts of the Everglades National Park (IUCN, 2012; 10.2305/IUCN.UK.2012-1.RLTS.T193451A2237271.en). As part of an invasive species monitoring program, Burmese pythons were captured in the Florida Everglades and used as a model for the development of sperm cryopreservation protocols for endangered snakes. Sperm was collected by flushing the vas deferens postmortem and initial motility score (IMS; % motile × speed of progression2), plasma membrane integrity (IPL), and acrosome integrity (IAC) were recorded before cryopreservation. Sperm was extended in TEST-yolk buffer with final dimethyl sulfoxide (DMSO) or glycerol (GLY) concentrations of 8, 12, or 16%, or combinations of DMSO and GLY with final concentrations of 4:4, 6:6, or 8:8%. Sperm in 500 µL of extender was frozen in vials at 0.3°C/min to –40°C before storage in liquid nitrogen. For each treatment, triplicate vials from each of 3 males were thawed at 37°C for 90 s. Cryoprotectant was removed by centrifugation and the sperm pellet was resuspended in TCM-199+HEPES. Sperm was evaluated at 22°C immediately following resuspension (T0) and at 60 (T60) minutes. All data were expressed as a percentage of initial (%IMS, %IPL, and %IAC). The effects of freeze method on %IMS, %IPL and %IAC were analysed by ANOVA and Tukey’s HSD test. Freeze method significantly affected %IMS at T0 (P = 0.0004) and T60 (P = 0.0001), with sperm frozen in the 6%DMSO:6%GLY and 4%DMSO:4%GLY treatments resulting in the highest %IMS at both T0 (19.4% and 17.7%, respectively) and T60 (26.7% and 14.4%, respectively). Regardless of cryoprotectant concentrations, sperm frozen in a combination of DMSO and GLY exhibited significantly higher %IMS than all treatments of DMSO or GLY alone (P < 0.0001 at T0 and T60). The %IPL was significantly affected by freeze method at T0 (P < 0.0001) and T60 (P = 0.0266). Sperm frozen in 8%DMSO:8%GLY and 6%DMSO:6%GLY retained greater %IPL at both T0 (69.1% and 65.7%, respectively) and T60 (47.8% and 49.9%, respectively). Acrosome integrity was significantly affected by freeze method at T0 (P < 0.0001) and sperm frozen in 8% DMSO resulted in the greatest %IAC (56.4%). In addition, all DMSO and DMSO:GLY treatments preserved a significantly greater proportion of intact acrosomes than GLY alone (P < 0.0001). To simplify these analyses and to determine the best overall freeze method for this species, a sperm quality index (SQI) was calculated, giving equal weight to each of the 3 measured indicators of cryosurvival. The SQI analysis revealed that Burmese python sperm frozen at 0.3°C/min in either 6%DMSO:6%GLY or 4%DMSO:4%GLY exhibited significantly higher post-thaw viability at T0 and T60 than all other treatments. This study represents the first comparative, comprehensive attempt to develop a sperm cryopreservation protocol for any snake species.

Different carbon sources affects biofloc volume, water quality and the survival and physiology of African catfish Clarias gariepinus fingerlings reared in an intensive biofloc technology system

A 6-week experiment was performed to compare different carbon sources, i.e. sucrose, glycerol and rice bran, to a nitrogen ratio of 15:1 in a biofloc-based African catfish Clarias gariepinus culture system. Catfish survival, growth, whole-body proximate composition, body indices, liver histopathology and glycogen content were measured. Each treatment was triplicated in glass aquaria with each replicate containing 50 fish (500 fish/m3) with an initial weight ± SD of 5.06 ± 0.05 g. Glycerol significantly increased total biofloc production, and both the sucrose and glycerol treatments generally had lower nitrogenous levels, compared to the control. These levels spiked at week 2 in the rice bran treatment, leading to significantly lower survival compared to all other treatments. At both weeks 3 and 6, liver histopathology of fish in the rice bran treatment revealed substantial vacuolation and less glycogen while the highest was in fish from the glycerol treatment. Fish growth was unaffected among the treatments, but survival was highest in the glycerol treatment. Rice bran appears unsuitable for C. gariepinus, likely due to being a slower-releasing carbon source. Instead, glycerol is recommended based on significantly higher biofloc production and subsequently improved water quality and survival of C. gariepinus during their nursery culture.

Effect of glycerol in combination with alfalfa on in vitro gas production and microbial protein synthesis

Alfonso-Ávila, Á. R., Charbonneau, E., Lafrenière, C. and Berthiaume, R. 2015. Effect of glycerol in combination with alfalfa on in vitro gas production and microbial protein synthesis. Can. J. Anim. Sci. 95: 577–588. This study sought to determine the effect of added glycerol on microbial protein synthesis, ruminal degradation and utilization of alfalfa at different concentrations of nonstructural carbohydrates (NSC), using in vitro gas production. The 2×3 factorial plus one treatment consisted of oven-dried alfalfa with two NSC levels [high: 17.9 (HNSC) or low: 7.4% dry matter (DM) (LNSC)] and three glycerol treatments [control without glycerol, 15% crude glycerol (CG) and 15% pure glycerol (PG)], the additional treatment was LNSC+exogenous sugars (LNSC+ES: LNSC with 5% sucrose+5% starch). Five pre-planned contrasts were evaluated from the seven treatments: (1) HNSC vs. LNSC alfalfa; (2) with glycerol vs. without; (3) interaction of alfalfa and glycerol; (4) CG vs. PG, and; (5) LNSC+ES vs. HNSC. Gas production over 24 h was higher for HNSC than LNSC (202 vs. 179 mL g−1 DM) and with glycerol than without glycerol (202.2 vs. 168 mL g−1 DM). A decrease in the acetate:propionate ratio was observed for HNSC compared with LNSC (2.87 vs. 3.27) and for the addition of glycerol vs. no glycerol (2.78 vs. 3.65). Reduced microbial mass (185.5 vs. 240.5 mg g−1 DM) was observed for CG compared with PG. The LNSC+ES treatment had lower microbial protein synthesis and propionic acid production in relation to HNSC. No significant interaction was observed between the effect of NSC content of alfalfa and glycerol utilization. When effects were studied separately, results indicate that increasing NSC in alfalfa stimulates the synthesis of microbial protein. Also, the addition of glycerol promotes the synthesis of glucose precursors. Finally, the type of glycerol has an impact on results obtained suggesting caution when extrapolating results for PG to CG.

Effects of Replacing Sucrose with Various Sugar Alcohols on Quality Properties of Semi-dried Jerky

The objective of this study was to investigate the effects of replacing sucrose with sugar alcohols (sorbitol, glycerol and xylitol) on the quality properties of semi-dried jerky. Total 7 treatments of jerkies were prepared as follows: control with sucrose, and treatments with 2.5 and 5.0% of sucrose replaced by each sugar alcohol, respectively. Drying yield, pH, water activity, moisture content, shear force, myofibrillar fragmentation index (MFI), 2-thiobarbituric acid reactive substance (TBARS) value, sugar content, and sensory evaluation were evaluated. Xylitol slightly decreased the pH when compared to the other sugar alcohols (p>0.05). The water activity of the semi-dried jerky was significantly reduced by treatment with glycerol and xylitol (p<0.05). The moisture content of semi-dried jerky containing various sugar alcohols was significantly higher than that of the control (p<0.05), while replacing sucrose with glycerol yielded the highest moisture content. The shear force of semi-dried jerky containing sugar alcohols was not significantly different for the sorbitol and glycerol treatments, but that replacing sucrose with 5.0% xylitol demonstrated the lowest shear force (p<0.05). The TBARS values of semi-dried jerkies with sugar alcohols were lower than the control (p<0.05). The sugar content of the semi-dried jerkies containing sorbitol and glycerol were lower than the control and xylitol treatment (p<0.05). In comparison with the control, the 5.0% xylitol treatment was found to be significantly different in the sensory evaluation (p<0.05). In conclusion, semi-dried jerky made by replacement with sugar alcohols improved the quality characteristics, while xylitol has applicability in manufacturing meat products.

Anti-irritant and anti-inflammatory effects of glycerol and xylitol in sodium lauryl sulphate-induced acute irritation.

Glycerol is known to possess anti-irritant and hydrating properties and previous studies suggested that xylitol may also have similar effects. Our aim was to study whether different concentrations of these polyols restore skin barrier function and soothe inflammation in sodium lauryl sulphate (SLS)-induced acute irritation. The experiments were performed on male SKH-1 hairless mice. The skin of the dorsal region was exposed to SLS (5%) for 3 h alone or together with 5% or 10% of glycerol respectively. Further two groups received xylitol solutions (8.26% and 16.52% respectively) using the same osmolarities, which were equivalent to those of the glycerol treatments. The control group was treated with purified water. Transepidermal water loss (TEWL) and skin hydration were determined. Microcirculatory parameters of inflammation were observed by means of intravital videomicroscopy (IVM). Furthermore, accumulation of neutrophil granulocytes and lymphocytes, the expression of inflammatory cytokines and SLS penetration were assessed, as well. Treatment with the 10% of glycerol and both concentrations of xylitol inhibited the SLS-induced elevation of TEWL and moderated the irritant-induced increase in dermal blood flow and in the number of leucocyte-endothelial interactions. All concentrations of the applied polyols improved hydration and prevented the accumulation of lymphocytes near the treatment site. At the mRNA level, neither glycerol nor xylitol influenced the expression of interleukin-1 alpha. However, expression of interleukin-1 beta was significantly decreased by the 10% glycerol treatment, while expression of tumour necrosis factor-alpha decreased upon the same treatment, as well as in response to xylitol. Higher polyol treatments decreased the SLS penetration to the deeper layers of the stratum corneum. Both of the analysed polyols exert considerable anti-irritant and anti-inflammatory properties, but the effective concentration of xylitol is lower than that of glycerol.

The effect of replacing barley with glycerol in the diet of dairy cows on rumen parameters and milk fatty acid profile

The objective of this paper was to evaluate the changes in rumen parameters (pH, NH3-N and volatile fatty acids (VFA) proportions), caused by large amounts of dietary glycerol fed to dairy cows, and whether those changes are associated with differences in the milk fatty acid (FA) composition. For this study eight primiparous Estonian Holstein cows (134±5 DIM at the beginning of the experiment) were paired and used in a 4×4 Latin square experimental design. Cows were fed a total mixed ration ad libitum, consisting of grass silage, barley meal, soybean meal and minerals (control) with crude glycerol isoenergetically replacing barley meal in experimental diets to inclusion levels of 52g/kg, 104g/kg and 156g/kg of crude glycerol in the dry matter (DM) of the diets. The effect of the diets was studied with the analyses of variance followed by the tests of linear and quadratic contrasts. Pearson correlation was used to assess the suitability of models predicting rumen VFA proportions based on milk FA, when glycerol-containing diets were fed. Results showed that dietary glycerol did not affect ruminal pH but linearly increased the relative proportions of propionate and butyrate in rumen fluid, at the expense of acetate. The ratios of acetate to propionate, and lipogenic VFA to propionate, showed both linear and quadratic changes. The proportion of 2-methyl butyrate changed both linearly and quadratically; highest when the diet with 52g/kg in DM of crude glycerol was fed. Rumen total VFA content decreased linearly according to the crude glycerol content in the diets. Replacing barley meal with glycerol in the diets affected the concentration of most of the de novo synthesized milk FA, as well as FA in different groups according to chain length or degree of saturation. The proportions of even-chain saturated FA C6:0–C10:0 increased linearly in milk fat of cows in the dietary glycerol treatments, whereas the proportions of C14:0 and C16:0 decreased quadratically. The proportion of saturated odd-chain FA (C5–C17) increased linearly and quadratically with increasing dietary glycerol treatments. Correlation analysis indicated that the increase in the proportion of propionate in total VFA was accordance with the changes in the milk FA profile, increasing the odd-chain (C5–C17) FA concentration.

Effects on enteric methane production and bacterial and archaeal communities by the addition of cashew nut shell extract or glycerol—An in vitro evaluation

The objective of the study was to evaluate the effect of cashew nut shell extract (CNSE) and glycerol (purity >99%) on enteric methane (CH4) production and microbial communities in an automated gas in vitro system. Microbial communities from the in vitro system were compared with samples from the donor cows, in vivo. Inoculated rumen fluid was mixed with a diet with a 60:40 forage:concentrate ratio and, in total, 5 different treatments were set up: 5mg of CNSE (CNSE-L), 10mg of CNSE (CNSE-H), 15mmol of glycerol/L (glycerol-L), and 30mmol of glycerol/L (glycerol-H), and a control without feed additive. Gas samples were taken at 2, 4, 8, 24, 32, and 48h of incubation, and the CH4 concentration was measured. Samples of rumen fluid were taken for volatile fatty acid analysis and for microbial sequence analyses after 8, 24, and 48h of incubation. In vivo rumen samples from the cows were taken 2h after the morning feeding at 3 consecutive days to compare the in vitro system with in vivo conditions. The gas data and data from microbial sequence analysis (454 sequencing) were analyzed using a mixed model and principal components analysis. These analyses illustrated that CH4 production was reduced with the CNSE treatment, by 8 and 18%, respectively, for the L and H concentration. Glycerol instead increased CH4 production by 8 and 12%, respectively, for the L and H concentration. The inhibition with CNSE could be due to the observed shift in bacterial population, possibly resulting in decreased production of hydrogen or formate, the methanogenic substrates. Alternatively the response could be explained by a shift in the methanogenic community. In the glycerol treatments, no main differences in bacterial or archaeal population were detected compared with the in vivo control. Thus, the increase in CH4 production may be explained by the increase in substrate in the in vitro system. The reduced CH4 production in vitro with CNSE suggests that CNSE can be a promising inhibitor of CH4 formation in the rumen of dairy cows.

Cryopreservation of turkey semen: Effect of breeding line and freezing method on post-thaw sperm quality, fertilization, and hatching

Cryopreservation methods for poultry semen are not reliable for germplasm preservation, especially for turkeys, where fertility rates from frozen/thawed semen are particularly low. The objective was to evaluate cryopreservation methods for effectiveness in promoting cryosurvival and post-thaw function of sperm from five turkey lines: one commercial line and four research (RBC1; E; RBC2; F) lines from Ohio State University (OSU). The model for cryopreservation was set up as a 2×2×2×5 design for cryoprotectant (glycerol or dimethylacetamide (DMA)), cryopreservation medium (Lake or ASG), method of dilution (fixed dilution volume versus fixed sperm concentration) and turkey line, respectively. The final cryoprotectant concentrations were 11% glycerol or 6% DMA. Thawed sperm were evaluated for plasma membrane integrity and quality, motility, acrosome integrity and, after artificial insemination, for egg fertility and hatchability. Commercial turkey hens were used for all fertility trials, regardless of semen source. Turkey sperm frozen with glycerol exhibited higher membrane integrity and membrane quality upon thawing than turkey sperm frozen with DMA although no differences in total motility, and only minimal differences in progressive motility, were detected among the eight cryopreservation treatments. Within line, fertility was affected by cryoprotectant, medium and dilution method, where the overall highest percentages of fertile, viable embryos (Day 7) occurred for the DMA/ASG/fixed sperm concentration method, while high percentages (15.8–31.5%) of fertile, non-viable embryos (Day 1–6) were observed for multiple cryopreservation methods, including two glycerol treatments. From a single insemination, the duration of true and viable fertility in all lines was 10–13weeks and 9–10weeks, respectively. The duration of hatchability was 4–6weeks after insemination for four of the turkey lines. The highest percentage of viable embryos was observed for the commercial line (9.5±2.4%), followed by the E line (5.3±1.3%), F line (3.7±2.0%) and RBC2 line (2.6±0.8%). For the RBC1 line, there was 100% embryonic death by Day 6 of incubation. Overall, better fertility results were obtained with the cryoprotectant DMA, the ASG diluent and fixed sperm concentration. However, the applicability of this method for preserving semen from research populations may be line dependent.

Chlorella mirabilis as a Potential Species for Biomass Production in Low-Temperature Environment

Successful adaptation/acclimatization to low temperatures in micro-algae is usually connected with production of specific biotechnologically important compounds. In this study, we evaluated the growth characteristics in a micro-scale mass cultivation of the Antarctic soil green alga Chlorella mirabilis under different nitrogen and carbon sources followed by analyses of fatty acid contents. The micro-scale mass cultivation was performed in stable (in-door) and variable (out-door) conditions during winter and/or early spring in the Czech Republic. In the in-door cultivation, the treatments for nitrogen and carbon sources determination included pure Z medium (control, Z), Z medium + 5% glycerol (ZG), Z medium + 5% glycerol + 50 μM KNO3 (ZGN), Z medium + 5% glycerol + 200 μM NH4Cl (ZGA), Z medium + 5% glycerol + 1 mM Na2CO3 (ZNC), Z medium + 5% glycerol + 1 mM Na2CO3 + 200 μM NH4Cl (ZGCA) and Z medium + 5% glycerol + 1 mM Na2CO3 + 50 μM KNO3 (ZGCN) and were performed at 15°C with an irradiance of 75 μmol m−2 s−1. During the out-door experiments, the night-day temperature ranged from −6.6 to 17.5°C (daily average 3.1 ± 5.3°C) and irradiance ranged from 0 to 2,300 μmol m−2 s−1 (daily average 1,500 ± 1,090 μmol m−2 s−1). Only the Z, ZG, ZGN, and ZGC treatments were used in the out-door cultivation. In the in-door mass cultivation, all nitrogen and carbon sources additions increased the growth rate with the exception of ZGA. When individual sources were considered, only the effect of 5% glycerol addition was significant. On the other hand, the growth rate decreased in the ZG and ZGN treatments in the out-door experiment, probably due to carbon limitation. Fatty acid composition showed increased production of linoleic acid in the glycerol treatments. The studied strain of C. mirabilis is proposed to be a promising source of linoleic acid in low-temperature-mass cultivation biotechnology. This strain is a perspective model organism for biotechnology in low-temperature conditions.

Measurement of Extracellular Ca2+ Influx and Intracellular H+ Efflux in Response to Glycerol and PEG6000 Treatments

Measurement of Extracellular Ca2+ Influx and Intracellular H+ Efflux in Response to Glycerol and PEG6000 Treatments

Greenhouse gas emissions and nutrient supply rates in soil amended with biofuel production by-products

Ethanol production results in distiller grain, and biodiesel produces glycerol as by-product. However, there is limited information on effects of their addition on evolution of N2O and CO2 from soils, yet it is important to enable our understanding of impacts of biofuel production on greenhouse gas budgets. The objective of this study was to evaluate the direct effects of adding wet distillers grain (WDG), thin stillage (TS), and glycerol at three rates on greenhouse gas emissions (N2O and CO2) and nutrient supply rates in a cultivated soil from the Canadian prairies. The WDG and TS application rates were: 100, 200, or 400 kg N ha−1, whereas glycerol was applied at: 40, 400, or 4,000 kg C ha−1 applied alone (G − N) or in a combination with 300 kg N ha−1 (G + N). In addition, conventional amendments of urea (UR) and dehydrated alfalfa (DA) were added at the same rates of total N as the by-products for comparative purposes. The production of N2O and CO2 was measured over an incubation period of 10 days in incubation chambers and Plant Root Simulator™ resin membrane probes were used to measure nutrient (NH 4 + -N, NO 3 − -N, and PO 4 −3 -P) supply rates in the soil during incubation. Per unit of N added, urea tended to result in the greatest N2O production, followed by wet distillers grain and thin stillage, with glycerol and dehydrated alfalfa resulting in the lowest N2O production. Cumulative N2O production increased with increasing the rate of N-containing amendments and was the highest at the high rate of UR treatment. Addition of urea with glycerol contributed to a higher rate of N2O emission, especially at the low rate of glycerol. The DA and WDG resulted in the greatest evolution of CO2 from the soil, with the thin stillage resulting in less CO2 evolved per unit of N added. Addition of N fertilizer along with glycerol enhanced microbial activity and decomposition. The amendments had significant impacts on release of available nutrient, with the UR treatments providing the highest NO 3 − -N supply rate. The TS treatments supplied the highest rate of NH 4 + -N, followed by WDG compared to the other amendments. The WDG treatments were able to provide the greatest supply of PO 4 −3 -P supply in comparison to the other amendments. Microbial N immobilization was associated with glycerol treatments applied alone. This study showed that the investigated biofuel by-products can be suitable soil amendments as a result of their ability to supply nutrients and N2O emissions that did not exceed that of the conventional urea fertilizer.

제주흑우 동결정액 제조 시 Amide 계열의 동결보호제가 동결 융해 후 정자의 성상에 미치는 영향

본 연구는 희소 한우인 제주흑우의 정액 동결을 위하여 동결보호제로서 glycerol과 amide 계열의 동결보호제인 5%의 DMA, DMF 그리고 7%의 MF를 이용하여 동결 융해 후 정자의 운동성, 생존율, 정자막 온전성 및 정자의 첨체 양상의 변화 조사를 위하여 수행하였다. 제주흑우의 정액을 동결 한 결과 glycerol과 DMF를 사용하였을 때 운동성은 <TEX>$64.00{\pm}9.62$</TEX>와 <TEX>$59.00{\pm}5.48$</TEX>로 DMA와 MF에 비해 유의적으로 높았으며(p<0.05), 생존율 역시 glycero과 DMF 처리구에서 <TEX>$58.25{\pm}6.63$</TEX>과 <TEX>$53.05{\pm}3.77$</TEX>로 유의적으로 높게 나타났다(p<0.05). 그러나 glycerol과 DMF 처리구 간의 유의적 차이는 나타나지 않았다. 정자막 온전성 검사(HOST)에 있어서도 역시 glycerol과 DMF 처리에서 정자 미부의 팽창 비율 역시 <TEX>$45.12{\pm}25.08$</TEX>과 <TEX>$44.95{\pm}8.58$</TEX>로 유의적으로 높았다(p<0.05). 정자의 첨체막 양상 변화에 있어 F pattern의 비율이 MF 처리시 다른 처리구에 비하여 유의적으로 낮았으며(p<0.05), B pattern의 비율은 DMA, DMF 및 MF의 사용시 glycerol에 비하여 유의적으로 증가하였다(p<0.05). AR pattern의 비율에 있어서는 DMF를 제외한 amide의 사용은 F pattern의 감소와 조기 첨체반응에 의한 유의적인 AR pattern의 증가를 볼 수 있었다(p<0.0.5). 그러나 DMF의 사용은 glycerol 보다 AR pattern의 비율이 유의적으로 낮았다(p<0.05). 동결 융해 후 시간에 따른 정자의 생존율과 정자미부 팽창 비율에 있어서 융해 초기에는 glycerol에서 DMF 보다 높은 생존율과 정자 미부 팽창율을 나타냈으나, 1시간 이후부터는 DMF에서 glycerol 보다 높은 생존율과 정자 미부 팽창율을 나타내었다. 이러한 결과는 희소 가축의 생식세포 보존 및 유전자원 확립을 위한 중요한 자료가 될 것이며, 제주흑우 및 희소 가축의 안정적이고 효율적인 동결 정액 제조 연구에 있어 중요한 정보를 제공할 것으로 평가된다. The objective of this study was to examine the effect of amides as a cryoprotectant for semen cryopreservation in Korean Jeju Black Bull. The semen was cryopreserved with extenders containing 5% dimethyl acetamide (DMA), 5% dimethyl formamide (DMF), 5% methyl formamide (MF) or 7% glycerol. Post-thawed sperm were evaluated for sperm motility, viability, acrosome integrity and membrane integrity. Post-thawed sperm motility was significantly higher (p<0.05) in glycerol and DMF (<TEX>$64.00%{\pm}9.62$</TEX> and <TEX>$59.00%{\pm}5.48$</TEX>, respectively) than DMA and MF (<TEX>$50.00%{\pm}3.24$</TEX> and <TEX>$44.00%{\pm}4.18$</TEX>, respectively). Sperm viability wassignificantly higher (p<0.05) in glycerol and DMF (<TEX>$58.25%{\pm}7.35$</TEX> and <TEX>$53.05%{\pm}3.77$</TEX>, respectively) than others. However, for sperm motility and viability, there were no differences among glycerol and DMF. Also, swelling sperm ratio by hypo-osmetic selling test (HOST) was significantly increased (p<0.05) in glycerol and DMF treatments (<TEX>$45.12%{\pm}25.08$</TEX> and <TEX>$44.95%{\pm}8.58$</TEX>, respectively). The percentage of capacitated sperm assessed by CTC staining, F pattern was lower (p<0.05) in DMF than others. B pattern was increased (p<0.05) in DMA, DMF and MF when compared with glycerol. AR pattern ratio was decreased (p<0.05) in glycerol and DMF when compared with DMA and MF. These results suggested that amides performed better and could be used as a cryoprotectant for semen freezing of Korean Jeju Black Bull.