1,720,964 research outputs found

    Effect of semen preparation on casa motility results in cryopreserved bull spermatozoa

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    Computer-assisted sperm analyzers (CASA) have become the standard tool for evaluating sperm motility and kinetic patterns because they provide objective data for thousands of sperm tracks. However, these devices are not ready-to-use and standardization of analytical practices is a fundamental requirement. In this study, we evaluated the effects of some settings, such as frame rate and frames per field, chamber and time of analysis, and samples preparations, including thawing temperature, sperm sample concentration, and media used for dilution, on the kinetic results of bovine frozen-thawed semen using a CASA. In Experiment 1, the frame rate (30-60 frame/s) significantly affected motility parameters, whereas the number of frames per field (30 or 45) did not seem to affect sperm kinetics. In Experiment 2, the thawing protocol affects sperm motility and kinetic parameters. Sperm sample concentration significantly limited the opportunity to perform the analysis and the kinetic results. A concentration of 100 and 50 x 10(6) sperm/mL limited the device's ability to perform the analysis or gave wrong results, whereas 5, 10, 20, and 30 x 10(6) sperm/mL concentrations allowed the analysis to be performed, but with different results (Experiment 3). The medium used for the dilution of the sample, which is fundamental for a correct sperm head detection, affects sperm motility results (Experiment 4). In this study, Makler and Leja chambers were used to perform the semen analysis with CASA devices. The chamber used significantly affected motility results (Experiment 5). The time between chamber loading and analysis affected sperm velocities, regardless of chamber used. Based on results recorded in this study, we propose that the CASA evaluation of motility of bovine frozen-thawed semen using Hamilton-Thorne IVOS 12.3 should be performed using a frame rate of 60 frame/s and 30 frames per field. Semen should be diluted at least at 20 x 10(6) sperm/mL using PBS. Furthermore, it is necessary to consider the type of chamber used and perform the analysis within 1 or 2 min, regardless of the chamber used.[...

    The effect of the chamber on kinetic results in cryopreserved bull spermatozoa

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    In this study, the effect of the chamber used for the automated analysis of sperm motility by a computer-assisted semen analysis system on sperm kinematics was evaluated, and the cause of this effect was also verified. Twenty-three bull semen batches were thawed, and semen was diluted, aliquoted and analysed with six different chambers, (three capillary-loaded chambers and three droplet (DR)-loaded chambers). For each chamber type, each sample was analysed in quadruplicate, and the reliability of the analysis was tested using an intraclass correlation coefficient (ICC). Furthermore, sperm membrane integrity (MI) was evaluated, for each sample and chamber, in 12 randomly selected central and 12 edge fields. The ICC analysis showed that some parameters could have a significant variability related to the chamber. High stability of results was detected in Leja 4-chamber slide. Furthermore, as previously reported in other studies, capillary-loaded chambers seemed to affect the total and progressive motility and sperm velocities (average path velocity, straight line velocity and curvilinear velocity). These findings were corroborated by the evaluation of sperm MI that was significantly higher in the DR-loaded chambers. This study confirms that the chamber used for the objective kinetic evaluation of bull-thawed spermatozoa significantly affects the result. These differences could be present also in other species, even if the specific effect on the sperm kinematics should be verified. The Makler chamber seemed to give reliable results with negligible effects on sperm kinematics

    Kinematic study on the effect of pH on bull sperm function

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    Since the mammalian spermatozoa became capable of motion, during the epididymal transit, the spermatozoon swims in a liquid medium and it is completely dependent on the environmental conditions. Some reports have suggested an influence of pH on sperm kinetic characteristics, but no study has objectively described how motility changes in a different environmental pH. In this study, we evaluated the effect of different environmental pHs (5.5, 6, 6.5, 7, 7.5, 8, and 8.5) on kinetic parameters, sperm viability, mitochondrial activity, and sperm morphology of bull semen immediately and 1h after dilution. The results showed higher values for sperm motility characteristics, viability, and mitochondrial activity at pH 7 and 7.5. Values of pH lower than 6.5 and higher than 8 resulted in suboptimal motility, with a decrease in most parameters. At pH 8 and 8.5, a discrepancy between viability and total and progressive motility was found, with a significant amount of spermatozoa that were live but immotile. This reduction seemed related to a decrease in mitochondrial activity, possibly due to the increase in pH. The flow cytometric evaluation of sperm viability assessed by calcein AM was very consistent with the amount of spermatozoa with membrane integrity, evaluated in fluorescence by propidium iodide/SYBR-14 stain. Thus, the calcein AM stain could be used as viability stain instead the classic propidium iodide/SYBR-14 stain because this could allow the addiction of other functional stains without a overlapping of the fluorescent signal in the flow cytometer.[...

    Cheesemaking in highland pastures: Milk technological properties, cream, cheese and ricotta yields, milk nutrients recovery, and products composition

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    Summer transhumance of dairy cows to high Alpine pastures is still practiced in many mountainous areas. It is important for many permanent dairy farms because the use of highland pastures increases milk production and high-priced typical local dairy products often boost farm income. As traditional cheese- and ricotta-making procedures in Alpine pastures are central to this dairy system, the objective of this study was to characterize the quality and efficiency of products and their relationships with the quality and availability of grass during the grazing season. The milk from 148 cows from 12 permanent farms reared on a temporary farm located in Alpine pastures was processed every 2 wk during the summer (7 cheesemakings from late June to early September). During each processing, 11 dairy products (4 types of milk, 2 by-products, 3 fresh products, and 2 ripened cheeses) were sampled and analyzed. In addition, 8 samples of fresh forage from the pasture used by the cows were collected and analyzed. At the beginning of the pasture season the cows were at 233 ± 90 d in milk, 2.4 ± 1.7 parities, and produced 23.6 ± 5.7 kg/d of milk. The milk yield decreased with the move from permanent to temporary farms and during the entire summer transhumance, but partly recovered after the cows returned to the permanent farms. Similar trends were observed for the daily yields of fat, protein, casein, lactose, and energy, as we found no large variations in the quality of the milk, with the exception of the first period of Alpine pasture. The somatic cell counts of milk increased during transhumance, but this resulted from a concentration of cells in a lower quantity of milk rather than an increase in the total number of cells ejected daily from the udder. We noted a quadratic trend in availability of forage (fresh and dry matter weight per hectare), with a maximum in late July. The quality of forage also varied during the summer with a worsening of chemical composition. The evening milk (before and after natural creaming), the whole morning milk, and the mixed vat milk had different chemical compositions, traditional coagulation properties, and curd-firming modeling parameters. These variations over the pasture season were similar to the residual variations with respect to chemical composition, and much lower with respect to coagulation and curd-firming traits. Much larger variations were noted in cream, cheese, and ricotta yields, as well as in nutrient recoveries in curd during the pasture season. The protein content of forage was correlated with some of the coagulation and curd-firming traits, the ether extract of forage was positively correlated with milk fat content and cheese yields, and fiber fractions of forage were unfavorably correlated with some of the chemical and technological traits. Traditional cheese- and ricotta-making procedures showed average cream, cheese, and ricotta yields of 6.3, 14.2, and 4.9%, respectively, and an overall recovery of almost 100% of milk fat, 88% of milk protein, and 60% of total milk solids

    Going Beyond Counting First Authors in Author Co-citation Analysis

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    The present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed

    Variations on the Author

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    “Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
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