A spermiogramme is an analysis performed to assess the quality of semen. Motility parameters are primarily obtained by subjectively observing samples or by using complex systems, such as computer-assisted sperm analysis (CASA). Here, we describe an easy and low-cost analysis system for obtaining quantifiable kinetic observations using Salmo salar semen model. In this work, we observed and captured video images of both fresh and stored Atlantic salmon semen to describe the possibility of analysis using the ImageJ CASA plug-in application for the kinetic parameters obtained from the videos. The semen is exposed to "powermilt", a commercial activating solution, and the curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP) values are described. When the samples were activated after having been stored, differences were detected in sperm quality, using the low-cost plug-in application. However, this system was not able to detect small variations in the same recorded sample, suggesting limits in sample observation. The results indicate that it is possible to quantify the kinetic parameters of semen samples using a low-cost video system and free software.
This study estimates spermatozoa mortality, morphology, motility and intracellular calcium levels in Atlantic salmon Salmo salar milt after prolonged storage. Milt samples were preserved at 4° C for 25 days and then evaluated for mortality. Motility remained high for the first 3 days and the mortality was low during the first 5 days of storage. A decrease of >50% in calcium content was observed after 5 days of storage. When spermatozoa were activated, calcium levels increased >200% in relative fluorescence units (RFU); this rate of increase was lost when the samples were stored for extended periods of time and was only partially manifested in a zero calcium solution. The results suggest that in vitro storage of S. salar spermatozoa at 4° C for a period of 3 days preserves motility and limits mortality to levels similar to those of fresh spermatozoa. This method also maintains intracellular calcium storage critical for spermatozoa performance.
Gastric distension has been used to evaluate gastric sensory function in humans, but the methodology is poorly validated and studies in vivo comparing different distension protocols are lacking. We aimed to compare the influence of the mode of gastric distension on sensation and gastric compliance utilizing a barostat device. In seven healthy volunteers, we positioned a barostat bag in the proximal stomach and tested in random order (in triplicate) four different distension protocols: (1) standard ramp distension with 4 mm Hg pressure step increments of 20 sec duration; (2) slow ramp distension with 2 mm Hg pressure increments of 40 sec duration; (3) random distension using a pressure ramp consisting of 2 mm Hg increments of 40 sec duration with randomly interposed pressure steps 50% below the preceding pressure step; and (4) rapid random distension with 4 mm Hg pressure increments of 10 sec duration with randomly interposed pressure steps 50% below the preceding pressure step. The distension procedures yielded mean airflow rates during the different distension protocols between 2.4 ml/sec for standard ramp and 18.4 ml/sec for rapid random distension. First perception and maximal tolerable pressure were 10.9 +/- 1.1 mm Hg and 19.6 +/- 1.5 mm Hg, respectively. First perception and maximal tolerable pressures were significantly correlated (r = 0.93, P < 0.005). The gastric pressure at occurrence of perception and the maximal tolerated pressure were not significantly different for the different distension protocols but gastric compliance was significantly reduced during rapid ramp distension (P < 0.01 vs slow ramp and P < 0.05 vs random distension) but not during standard ramp distension. We conclude that gastric sensory pressure thresholds as assessed by isobaric distension are not influenced by the mode of distension. The high correlation of pressure thresholds at first perception and maximal tolerated distension suggest a single population of gastric mechanoreceptors that mediate first sensation at low intensity stimulation and pain at intense stimulation.
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