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      Evaluación de cámaras de recuento sobre parámetros espermáticos de verracos analizados con un sistema CASA-Mot Translated title: Assessment of counting chambers on boar sperm parameters analyzed by a CASA-Mot system


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          Resumen Introducción. La comprensión de la variabilidad en los valores de la cinética de los espermatozoides, por medio de diferentes profundidades de cámaras, demuestra la importancia de crear un estándar para los métodos de control de calidad en la industria de la inseminación artificial (IA). Objetivo. El objetivo del presente trabajo fue evaluar los parámetros de cinética de los espermatozoides con base en diferentes profundidades de cámara de visualización mediante un sistema comercial de análisis de esperma asistidos por computadora, CASA-Mot. Materiales y Métodos. Se utilizaron veinte dosis seminales de diez verracos pietrain. El periodo experimental fue de febrero a julio del año 2017. Se utilizó el sistema integrado de análisis de semen (ISAS®v1) con una frecuencia de captura de 50 Hz. Se utilizaron cámaras de recuento ISAS®D4C16 e ISAS®D4C20 con una altura de 16 y 20 μm, respectivamente y precalentadas a 37 °C. Resultados. Se encontraron valores superiores (p<0,05) para todos los parámetros cinéticos cuando la altura de la cámara de recuento fue de 20 μm. El efecto de zona dentro de la cámara de recuento se mantuvo constante entre las dos alturas, y las variaciones observadas en los parámetros cinéticos se debieron a un efecto aleatorio del verraco. Al analizar el efecto de zona dentro de la cámara de recuento, los primeros tres campos de análisis mostraron mayor velocidad curvilínea y rectilínea (p<0,05) que los siguientes campos, lo que se atribuye a la presencia de movimiento pasivo (drifting). Conclusión. La mayor amplitud y capacidad de volumen dentro de la cámara de recuento (20 μm versus 16 μm), podría favorecer el desplazamiento sin restricciones de las células, lo que explicaría el incremento en los valores cinéticos conforme aumentó la altura de la cámara. Deben continuarse los estudios sobre las condiciones técnicas del análisis seminal para estandarizar los métodos de valoración con los sistemas CASA.

          Translated abstract

          Abstract Introduction. Understanding the variability in sperm kinetic values through different chambers depths, shows the importance to create a standard for quality control methods in the artificial insemination (AI) industry. Objective. The work aimed was to evaluate the spermatozoa kinetic parameters based on different depths of the visualization chamber by means of a commercial system of computer-assisted sperm analysis, CASA-Mot. Materials and Methods. Twenty seminal doses of ten pietrain boars were used. The experimental period was from February to July 2017. The Integrated Semen Analyses System (ISAS®v1) with 50 Hz capture frequency was used. ISAS®D4C16 and ISAS®D4C20 counting chambers with a height of 16 and 20 μm respectively and pre-heated to 37 °C were employed. Results. Higher values (p<0.05) were found for all kinetic parameters when the height of the counting chamber was 20 μm. The zone effects within the counting chamber were constant between the two heights, and the variations observed in the kinetic parameters were due to a random effect of the boar. When analyzing the zone effect within the counting chamber, the first three fields of analysis showed higher curvilinear and rectilinear velocity (p<0.05) than the following fields, which is attributed to the presence of passive movement (drifting). Conclusion. The greater amplitude and volume capacity within the counting chamber (20 μm versus 16 μm), could promote the unrestricted movement of the cells, which would explain the increase in the kinetic values as the chamber height increased. Studies on the technical conditions of seminal analysis should be continued in order to standardize valuation methods with CASA systems.

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          Computer assisted semen analyzers in andrology research and veterinary practice.

          The evaluation of sperm cell motility and morphology is an essential parameter in the examination of sperm quality and in the establishment of correlations between sperm quality and fertility. Computer-assisted sperm analysis (CASA) allows an objective assessment of different cell characteristics: motion, velocity, and morphology. The development and problems related to this technology are raised in this review, paying particular attention to the biases and standardization requirements absolutely needed to obtain useful results. Although some interesting results, mainly in humans, have already been obtained, many questions remain, which have to be answered to allow for further development of this technology in veterinary medicine, clinical fertility settings, physiological, and toxicology research activities. The main problem is related to the standardization and optimization of the equipment and procedures. The different CASA instruments have all demonstrated high levels of precision and reliability using different sperm classification methodology. Their availability gives us a great tool to objectively compare sperm motility and morphology and to improve our knowledge and ability to manipulate spermatozoa.
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            Computer-assisted sperm analysis (CASA): capabilities and potential developments.

            Computer-assisted sperm analysis (CASA) systems have evolved over approximately 40 years, through advances in devices to capture the image from a microscope, huge increases in computational power concurrent with amazing reduction in size of computers, new computer languages, and updated/expanded software algorithms. Remarkably, basic concepts for identifying sperm and their motion patterns are little changed. Older and slower systems remain in use. Most major spermatology laboratories and semen processing facilities have a CASA system, but the extent of reliance thereon ranges widely. This review describes capabilities and limitations of present CASA technology used with boar, bull, and stallion sperm, followed by possible future developments. Each marketed system is different. Modern CASA systems can automatically view multiple fields in a shallow specimen chamber to capture strobe-like images of 500 to >2000 sperm, at 50 or 60 frames per second, in clear or complex extenders, and in <2 minutes, store information for ≥ 30 frames and provide summary data for each spermatozoon and the population. A few systems evaluate sperm morphology concurrent with motion. CASA cannot accurately predict 'fertility' that will be obtained with a semen sample or subject. However, when carefully validated, current CASA systems provide information important for quality assurance of semen planned for marketing, and for the understanding of the diversity of sperm responses to changes in the microenvironment in research. The four take-home messages from this review are: (1) animal species, extender or medium, specimen chamber, intensity of illumination, imaging hardware and software, instrument settings, technician, etc., all affect accuracy and precision of output values; (2) semen production facilities probably do not need a substantially different CASA system whereas biology laboratories would benefit from systems capable of imaging and tracking sperm in deep chambers for a flexible period of time; (3) software should enable grouping of individual sperm based on one or more attributes so outputs reflect subpopulations or clusters of similar sperm with unique properties; means or medians for the total population are insufficient; and (4) a field-use, portable CASA system for measuring one motion and two or three morphology attributes of individual sperm is needed for field theriogenologists or andrologists working with human sperm outside urban centers; appropriate hardware to capture images and process data apparently are available.
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              The lateral migration of a spherical particle in two-dimensional shear flows


                Author and article information

                Role: ND
                Role: ND
                Agronomía Mesoamericana
                Agron. Mesoam
                Universidad de Costa Rica. Programa Cooperativo Centroamericano para el Mejoramiento Cultivos y Animales (San Pedro, San José, Costa Rica )
                August 2019
                : 30
                : 2
                : 447-458
                [1] orgnameInstituto Tecnológico de Costa Rica Costa Rica
                [2] orgnameInstituto Tecnológico de Costa Rica Costa Rica

                This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 International License.

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                Figures: 0, Tables: 0, Equations: 0, References: 40, Pages: 12

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