The Andean puma (Puma concolor) has not been widely studied, particularly in reference to its semen characteristics. The aim of the present study was to define the morphometry of puma sperm heads and classify their subpopulations by cluster analysis. Samples were recovered postmortem from two epididymides from one animal and prepared for morphological observation after staining with the Hemacolor kit. Morphometric data were obtained from 581 spermatozoa using a CASA-Morph system, rendering 13 morphometric parameters. The principal component (PC) analysis was performed followed by cluster analysis for the establishment of subpopulations. Two PC components were obtained, the first related to size and the second to shape. Three subpopulations were observed, corresponding to elongated and intermediate-size sperm heads and acrosomes, to large heads with large acrosomes, and to small heads with short acrosomes. In conclusion, puma spermatozoa showed no uniform sperm morphology but three clear subpopulations. These results should be used for future work in the establishment of an adequate germplasm bank of this species.
Semen analysis is a key factor when determining the fertility ability in males. South American camelids, and in particular the alpaca, have been studied very little when compared with other farm animals. The aim of this work was to perform the kinematic characterization of alpaca spermatozoa collected directly from the deferent duct by using CASA-Mot (Computer Assisted Semen Analysis for Motility) technology. Samples were obtained every three days throughout the reproductive season during two periods and with a break of seven days in the middle. During both periods, the quality of the sample's motility and kinematics increased over the first two days and then subsequently decreased. This pattern was similar in all animals. It was concluded that the introduction of resting times can be useful to improve sperm quality for artificial insemination purposes in natural conditions.
Introducción. El estudio de la morfometría espermática del venado cola blanca (Odocoileus virginianus peruvianus) permite caracterizar las células espermáticas de esta especie con propósitos de conservación. Objetivo. Determinar las subpoblaciones espermáticas según la morfometría de los espermatozoides epididimarios del venado de cola blanca, con un sistema CASA-Morph. Materiales y métodos. El estudio se desarrolló en el segundo semestre 2016, en la Universidad Nacional de San Antonio Abad del Cusco, Perú. En muestras de semen obtenidas del extremo caudal del epidídimo de dos venados (dos y cuatro dientes), se determinó con base en el sistema CASA-Morph, Integrated Semen Analysis System (ISAS®v1): la longitud, anchura, área, perímetro, elipticidad, elongación, regularidad y rugosidad de la cabeza del espermatozoide, también, la anchura, área, distancia y ángulo de inserción de la pieza intermedia. Se realizó un análisis de componentes principales (PCA) y se aplicó el modelo no jerárquico k-medias para determinar el número óptimo de clústeres. Resultados. Las variables morfométricas se distribuyeron en cinco PCA: elipticidad, tamaño, circularidad, ángulo de inserción de la pieza intermedia y anchura de la pieza intermedia, que explicaron un 81,24 % de la varianza total. El análisis de clústeres determinó cuatro subpoblaciones (SP), la SP1 agrupó células grandes, alargadas con alta elongación y elipticidad (21,76 %), la SP2 compuesta por espermatozoides de tamaño pequeño y delgados tanto en la cabeza como en la pieza intermedia (37,7 %), la SP3 se relacionó con la presencia de células alargadas con valores altos de anchura de la pieza intermedia, elipticidad y elongación (17,7 %), y la SP4 agrupó espermatozoides de tamaño intermedio de la cabeza y de la pieza intermedia (22,84 %). Conclusiones. La determinación de cuatro subpoblaciones espermáticas en el semen de venado cola blanca puede ser la base de programas de reproducción asistida de esta especie.
The aim of the study was to determine the morphological and morphometric characteristics of the vicuña sperm (Vicugna vicugna), looking for possible subpopulations in its structure. Semen was collected by electroejaculation method from three adult male vicuñas weighing 50.33 ± 2.52 kg, once per animal. The volume, color and concentration, sperm morphology and morphometry were determined. The samples for the analysis of sperm morphology and morphometry were stained with Hemacolor®, and analyzed using the CASA-Morph, Integrated Semen Analysis System (ISAS®v1). Five forms of the vicuña sperm head were distinguished: normal, piriform, long, short and rounded. The morphometric parameters determined were the length, width, area, perimeter, ellipticity, elongation, regularity and rugosity of the vicuña sperm head, percentage of acrosome, head gray level, as well as the width, area, distance and angle of insertion of midpiece of the sperm. In relation to morphology, significant differences (P <0.05) were found in the percentage distribution of head shapes, with the normal shape (55.7%) being the majority and different from the other shapes. Significant differences (P <0.05) were found between animals in the morphometric variables of head, percentage of acrosome, ellipticity, rugosity, elongation and gray level; while the variables of the midpiece and regularity did not show differences (P> 0.05). The morphometric variables were distributed in four main components (PCA) called elongation, area, circularity and midpiece width, which explained 84.59% of the total variance. The cluster analysis determined five subpopulations (SP): SP1 grouped small cells of low length, width and area (18.8%); SP2 of sperm of large size, both in area and width of the head (17.38%); SP3 of rounded cells with high values of percentage of acrosome and head gray level (24.04%); SP4 of spermatozoa of intermediate size and elongated, with greater ellipticities and elongation (23.61%); and SP5 of cells of intermediate size and short, with low values of area and length (16.71%).
El objetivo del estudio fue determinar la existencia de subpoblaciones espermáticas según la morfometría de los espermatozoides de llama, utilizando un sistema CASA-Morph. Se colectó semen de cuatro llamas Q’ara de 4-6 años mediante el método de electroeyaculación en cuatro oportunidades por animal, con intervalos de una semana. Se determinó el volumen, movilidad total, concentración de espermatozoides, porcentaje de espermatozoides vivos y porcentaje de espermatozoides con reacción acrosomal. Los parámetros morfométricos de los espermatozoides se evaluaron utilizando el sistema CASA-Morph, Integrated Semen Analysis System (ISAS®v1). Se determinó la longitud, anchura, área, perímetro, elipticidad, elongación, regularidad y rugosidad de la cabeza del espermatozoide de llama, así como la anchura, área, distancia y ángulo de inserción de la pieza intermedia del espermatozoide. Las variables morfométricas se distribuyeron en cinco componentes principales (PCA) denominados elongación, longitud, circularidad, anchura de la pieza intermedia e inserción de la pieza intermedia, que explicaron un 81.6% de la varianza total. El análisis de clústeres determinó cuatro subpoblaciones (SP): SP1 agrupó células pequeñas con baja elongación y elipticidad (36.8%); SP2 de espermas de tamaño intermedio, tanto en la longitud y anchura de la cabeza (9.9%); SP3 de células alargadas con valores bajos de anchura de la cabeza y pieza intermedia (33.0%); y SP4 de espermatozoides con una anchura de la pieza intermedia alta (20.2%). Se hallaron diferencias significativas de subpoblaciones intra e inter animal.
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