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El artículo expone parte de los resultados del proyecto de investigación «Violencia y endeudamiento: mecanismos que coartan los derechos humanos de personas endeudadas en el sector informal costarricense», desarrollado en la Universidad Estatal a Distancia. Bajo un carácter descriptivo, a partir de la revisión de antecedentes nacionales e internacionales y la participación de seis mujeres, se logra comprender el impacto del endeudamiento informal en la historia de vida de la población participante. Como parte de los hallazgos, las personas participantes evidenciaron una serie de privaciones relacionadas con la educación, empleo formal y conocimientos financieros; a su vez, la figura del crédito informal se destina a gastos regulares requeridos para la reproducción de condiciones de vida (pago de alquiler, alimentación o servicios básicos), con montos catalogados como «micro créditos», sujetos a plazos y condiciones definidos a criterio de la persona prestamista; espacio en el cual, se evidencian mecanismos coercitivos de presión y violación a la integridad de la persona (amenazas, daño físico) para pagar la deuda.

This investigation aimed to analyze the effect that thawing time and temperature in combination with a termo-resistance test had on straws from dairy bulls used for artificial insemination (AI) on semen motility and kinematic variables measured with CASA systems. Eight animals of Holstein and Jersey breeds were used, and nine frozen-thawed semen doses per animal were analyzed for each breed. Three temperatures (35, 37, and 40 °C) and three thawing times (35, 40, and 45 s) were evaluated using a factorial design. Motility and kinematic patterns were analyzed using CASA-mot (Computer-Assisted Semen Analysis of motility) technology at different post-thawing times (0.5, 1, and 2 h). Sperm motility in Jersey bulls was higher (p < 0.05) than in Holstein ones (64.52 ± 1.45% and 53.10 ± 1.40%, respectively). The same effect was seen with progressive motility among the two breeds (Jersey: 45.29 ± 1.00%; Holstein: 36.30 ± 0.98%, p < 0.05). The Jersey breed presented higher values (p < 0.05) of curvilinear velocity (VCL), rectilinear velocity (VSL), average velocity (VAP), linearity on forward progression (LIN), and wobble (WOB). The Holstein breed showed a lower mean value (p < 0.05) of the beat-cross frequency (BCF) compared to the Jersey breed, thus suggesting an effect on VCL and VAP. During the post-thaw period, a gradual increase in VCL was observed at 2 h. VSL and VAP showed a decrease (p < 0.05) as the post-thaw period was prolonged. The study showed differences in sperm quality between Holstein and Jersey breeds, influenced by cryopreservation, thawing, and post-thawing incubation. Thawing at 37 °C for 30 s was considered optimal in relation to sperm motility. In addition, a decrease in sperm quality was observed as post-thawing time increased.

Abstract Enzootic bovine leukosis virus (BLV) and bovine herpesvirus 1 (BHV-1) are very important infectious agents for the livestock industry worldwide. The present study aimed to explore the association between natural exposure to BLV and BHV-1 with sperm quality analyzed by Computer-Assisted Semen Analysis (CASA) systems. Ten sexually mature Brahman bulls, with sanitary status BLV+/BHV-1+ (n = 2), BLV−/BHV-1+ (n = 6) and BLV-/BHV-1- (n = 2) were evaluated twice, 30 days apart. Results showed that sanitary status of each bull was not associated with semen quality. It was found that the quality of the semen from the second collection was better due to the interruption of sexual rest. The evidence thus revealed that a bull infected with BLV generated good-quality contaminated semen and, therefore, that it is essential to detect contaminated seminal samples to prevent the spread of BLV. A multivariate analysis showed the presence of four sperm subpopulations in Brahman bulls that differ significantly in their kinematic patterns and with respect to sanitary status (P

Ethical considerations regarding our treatment of animals have gained strength, leading to legislation and a societal focus across various disciplines. This is a subject of study within curricula related to agri-food sciences. The aim was to determine the perceptions of agronomy university students concerning animal welfare in livestock production systems. A survey was conducted to encompass various aspects, from participants’ sociodemographic attributes to their attitudes and behaviors regarding animal welfare and the consumption of animal products. Statistical analysis, performed using R software, delved into the associations between participants’ characteristics and their perspectives on the ethical, bioethical, and legal dimensions of animal welfare. Associations between demographic factors and ethical viewpoints among students were identified. Gender differences emerged in animal treatment perceptions, while rural and urban environments impacted perspectives on various animals. Bioethical considerations revealed distinctive disparities based on gender and education in concerns regarding animal welfare, value perceptions, evaluations of animal behaviors, and opinions on animal research. It is crucial to distinguish between animal welfare and the ethical considerations arising from coexisting with sentient beings capable of experiencing suffering. Ethical theories provide a lens through which we perceive our obligations toward animals. The responsibility to ensure animal welfare is firmly rooted in recognizing that animals, like humans, experience pain and physical suffering. Consequently, actions causing unjustified suffering or mistreatment, particularly for entertainment purposes, are considered morally unacceptable.

Introducción. Los patrones cinemáticos del semen se condicionan por variación individual y/o un efecto ambiental determinado por el manejo posterior a la obtención de la muestra. Objetivo. Evaluar el efecto de dos diluyentes comerciales, la edad y estado sexual de los toros sobre la movilidad y cinética espermática del semen fresco bovino. Materiales y métodos. El estudio se realizó en las fincas ganaderas “La Vega”, San Carlos, y “La Balsa”, San Ramón, Costa Rica, entre marzo y agosto de 2021. Se utilizó el semen proveniente de trece toros Brahman, extraído por electroeyaculación. Se utilizaron 45 eyaculados en total, con un mínimo de dos y un máximo de cinco por animal. El semen se diluyó con dos diluyentes comerciales, Optixcell® y Bioxcell®, a una temperatura de 37 °C. Las muestras se analizaron mediante el Sistema Computarizado de Análisis Seminal (Computer-Assisted Semen Analysis, CASA)-Mot ISAS®v1 y una cámara de recuento espermática reutilizable Spermtrack® de 20 μm de profundidad. Resultados. Se observaron diferencias (p48 meses (62,27±1,87 %). Se observó un efecto del estado sexual, durante la monta se presentó mayor linealidad, velocidad y oscilación espermática comparado al descanso sexual. Conclusión. La edad, el estado sexual y el diluyente utilizado para la preservación del semen condicionaron la calidad espermática de los eyaculados bovinos.

The objective of the present study was to determine the effects that the reproductive season has on the motility, kinematics, morphology, and sperm morphometry of Brahman bulls evaluated with a commercial CASA system. The experiment was carried out at the Costa Rica Institute of Technology from March to August 2021. A total of eight Brahman bulls were used. A total of 28 ejaculates were collected in the pre-mating period (PMP), during it (DMP), and after it (AMP) using an electroejaculator. The sperm concentration was measured with the Accuread photometer. The motility was measured using a Spermtrack® counting chamber. The analyses were performed with the CASA-Mot ISAS®v1 system. The morphology was analyzed using a microscope with a negative phase contrast objective. Morphometry was evaluated with the CASA-Morph. The sperm concentration did not present differences between the PMP and AMP; however, it was significantly higher than DMP (p > 0.05). Regarding the progressiveness variables, linearity on forward progression (LIN), straightness (STR), and wobble (WOB) were higher (p < 0.05) DMP. A kinematic principal component analysis grouped all the variables into three factors and an effect on the reproductive period was found (p < 0.05) in the parameters of the head and middle part of the sperm, such as width and perimeter, which were greater in the PMP. The length of the sperm head in the PMP and DMP did not show differences; however, both were larger (p < 0.05) than AMP. The insertion distance of the middle piece of the sperm was significantly greater than DMP. Finally, the PMP contained cells with a larger insertion angle (p < 0.05) than AMP. These findings are important to understand the implications of reproductive status on sperm quality and to consider them in andrological evaluations.

Introducción. La reproducción asistida de animales tiene su origen en la domesticación de las especies ganaderas útiles al ser humano. Los consiguientes procesos de selección artificial permitieron desarrollar técnicas biotecnológicas que contribuyeron a la mejora de las capacidades de producción animal. Objetivo. Contextualizar el análisis seminal en la agricultura de precisión en el siglo XXI. Desarrollo. La visualización de espermatozoides en el microscopio puede considerarse como el primer paso en el advenimiento de la biotecnología de la reproducción y la base para el desarrollo de las técnicas de reproducción asistida. Con el perfeccionamiento de los métodos de microscopía, se logró caracterizar los gametos masculinos, lo que significó un avance significativo en la tecnología de inseminación artificial. El punto de inflexión marcado por el desarrollo de técnicas de conservación espermática, implicó un cambio sustantivo en el desarrollo de estas tecnologías en las especies ganaderas, ya sea con semen crio preservado o refrigerado. Estos métodos son de alto valor en el caso de especies amenazadas, dado que se pueden crear bancos de germoplasma con propósitos de conservación genética y rescate de especies en riesgo de extinción. El análisis seminal se ha desarrollado de forma paralela con las técnicas de reproducción asistida, al punto que hoy se considera como una técnica relevante en la biotecnología de la reproducción animal, que se ha perfeccionado mediante el avance de la ciencia y la tecnología, la física óptica y la computación. Conclusión. El análisis seminal ha tenido un cambio de paradigma al desechar técnicas obsoletas de evaluación subjetiva de la calidad seminal y adoptar métodos objetivos de evaluación del semen, mediante el análisis de grandes volúmenes de datos y variables de movilidad, cinéticas, morfométricas, morfológicas y fragmentación del ADN, que permiten caracterizar mejor los eyaculados de los reproductores en centros de inseminación artificial.

La suplementación vitamínica es importante para optimizar el potencial reproductivo del verraco. El objetivo de este estudio fue evaluar la calidad seminal del eyaculado de verracos suplementados con vitaminas liposolubles y el efecto de la durabilidad de las muestras de semen a través del tiempo. Se utilizaron cuatro verracos de una línea comercial terminal (Duroc*Pietrain) con edades promedio de 25,3±8,8 meses y de fertilidad conocida. Las muestras de semen se recogieron por la mañana, una vez por semana. Los animales se asignaron aleatoriamente a dos grupos y se alimentaron con cualquiera de los dos tratamientos durante 16 semanas: (1) tratamiento de control (TC, sin suplementos de vitaminas liposolubles), (2) tratamiento que contenía un 100 % de suplementos de vitaminas liposolubles (TS) a base de vitaminas liposolubles (A, D3, E). Se encontraron diferencias (P0,05) en los valores de movilidad total y progresiva a las 24 y 48 horas, pero si hubo una disminución significativa (P

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). 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%).

Ejaculate quality can be regarded as multifactorial, with nutrition being a factor that could directly influence sperm parameters. The present study aimed to evaluate seminal quality associated with seasonal fat-soluble vitamin supplementation of boars. Seven sexually mature boars were randomly allotted to one of the three groups, and fed one of the three supplementary diets for 32 weeks: (1) control treatment (COD), without supplementation of fat-soluble vitamins, (2) treatment containing 100% fat-soluble vitamin supplementation administered intramuscularly, which was based on fat soluble vitamin supplementation (A, D3, E) (FVD1), and (3) treatment containing 50% of fat-soluble vitamin supplementation (FVD12). Semen was collected at 7-day intervals. Semen samples were analyzed to assess several sperm parameters using the Computer-Assisted Semen Analysis (CASA) ISAS®v1 system. Results showed that groups receiving FVD1 and FVD12 supplementation had an increased semen volume. The percentages of motile and progressively motile sperm were increased by FVD1 treatment. A statistically significant interaction between treatment and season was found in the percentage of motility and progressive motility (p < 0.05). Sperm concentrations showed significant differences (p < 0.05) between treatments. Velocity variables (VSL, VCL, and VAP) were higher (p < 0.05) in boars that received fat-soluble vitamin supplementation in comparison to controls receiving no supplementation. The FVD1 treatment presented spermatozoa with greater head size and more elongated heads (p < 0.05). Overall, the utilization of dietary fat-soluble vitamin supplementation significantly improved the semen quality of boar ejaculates. This highlights the importance of fat-soluble vitamin supplementation in sexually active boars.

Artificial insemination in the swine industry, as in other species, demands adequate semen handling and accurate evaluation for the preparation of seminal doses. Sperm concentration and motility estimates are part of the semen evaluation process and are considered important for maximizing the yield of doses for insemination. In this study, methods were examined for their accuracy in the estimation of boar sperm concentration and motility. Assessments of sperm concentration were carried out using iSperm®, ISAS® v1, Open CASA v2, and the Accuread® photometer. Analyses of sperm motility were performed with iSperm®, ISAS® v1, and Open CASA v2 systems. In this study, boar semen samples were collected from 10 healthy males from two genetic lines. There were no relevant differences between sire lines when sperm concentration was assessed. A Bayesian analysis was applied to the four methods used to assess sperm concentration to examine whether there are relevant differences between them. Results suggested differences in the four methods, with a probability of relevance (PR) of 0.86–1.00. The iSperm® method revealed higher concentration values within the highest posterior density region at 95% confidence interval (HPD95%) = 167.0, 224.2 M/mL, whereas Open CASA v2 showed the lowest values, with HPD95% = 99.3, 155.9 M/mL. The iSperm® demonstrated higher reliability in measuring sperm concentration compared to other methods or devices within the given range of confidence. ANOVAs revealed relevant differences in the three methods of motility estimation. Overall, differences in boar sperm concentration and motility estimates were found using various methods, but further studies are needed for better characterization of these differences.

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.

Introducción. Las condiciones óptimas de análisis seminal permiten estandarizar los protocolos de evaluación. Objetivo. Evaluar el efecto de factores externos relacionados con el seminograma sobre las subpoblaciones espermáticas de bovinos de la raza Brahman. Materiales y métodos. El estudio se realizó con animales de dos fincas ganaderas de Alajuela, Costa Rica, de septiembre a diciembre de 2019. Se utilizaron diez toros Brahman que fueron electroeyaculados y el semen se diluyó con tres diluyentes comerciales: Andromed®, Androstar® y BTS, a dos temperaturas (37 y 29 °C). Las muestras se analizaron mediante un sistema CASA-Mot ISAS®v1 y se utilizaron cámaras de recuento ISAS®D4C (10, 16 y 20 μm) y Spermtrack® en diferentes tiempos de análisis (0, 3, 6 y 12 h). Resultados. La Spermtrack® presentó mayor velocidad curvilínea pero menor linealidad respecto de las otras cámaras de recuento (p

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

El análisis de la concentración espermá- tica en los eyaculados de verraco constituye una estimación de calidad en granjas dedicadas a la venta de dosis seminales e inseminación artifi- cial. El objetivo del presente estudio fue determi- nar las diferencias en precisión entre métodos de estimación de la concentración espermática. Se analizaron 174 eyaculados de verracos Duroc y Pietrain, pertenecientes a la granja del Programa de Producción Agropecuaria (PPA) de la Escuela de Agronomía del Instituto Tecnológico de Costa Rica. La concentración fue estimada por 3 méto- dos: espermadensímetro (ED), espectrofotómetro (EF) y sistema automatizado de análisis de semen (casa). La concentración más baja (p

Introducción. El uso de biotecnologías reproductivas ha significado una mejora relevante sobre los parámetros de eficiencia y rentabilidad en la industria porcina. Objetivo. Determinar el estado del arte en el que se encuentra la biotecnología aplicada al estudio del semen y la implementación; en granjas porcinas; de técnicas de reproducción asistida como la inseminación artificial. Desarrollo. En el sector porcino, la fertilidad de los verracos (Sus scrofa domestica) tiene un efecto significativo sobre la eficiencia general de la reproducción de las piaras. Aunque hay métodos objetivos de valoración precisa de las variables seminales de los verracos, muchas granjas aún continúan realizando sus evaluaciones de forma subjetiva. Los conceptos bioquímicos que determinan la funcionalidad del gameto masculino o espermatozoide son molecularmente complejos, sin embargo, pueden conocerse los aspectos generales de la funcionalidad y desarrollar métodos de manejo del semen que permita el mantenimiento de las condiciones óptimas de las muestras seminales o dosis de semen utilizadas en inseminación artificial (IA). Dentro de la funcionalidad espermática intervienen parámetros clásicos de volumen, concentración y movilidad que son fundamentales para la optimización de las dosis seminales producidas por verraco. Los sistemas CASA (Computerassisted semen analysis) son una aplicación biotecnológica que desplaza el análisis subjetivo tradicional del semen para la valoración de la calidad, estos aportan grandes volúmenes de información que han permitido explorar más en detalle los eyaculados porcinos. Conclusiones. En las granjas que utilizan la inseminación artificial, el uso de técnicas biotecnológicas para el análisis seminal, constituye un valor agregado para el estudio de la calidad del semen y la optimización de dosis seminales producidas.

This study aimed to characterize the sperm kinematic values with high frames per second, to define the subpopulation structure of a horse and a donkey and compare them. A total of 57 fresh semen ejaculates (26 Spanish and 16 Arabian horse breeds and 10 donkeys) were collected and subsequently analyzed for kinematic parameters using the Computer-aided sperm motility analysis ISAS®v1.2 system and using a Spermtrack® 10-μm depth counting chamber. Sequences were recorded at 250 frames per second, and eight kinematic parameters were automatically evaluated. All kinematic parameters showed significant differences between a donkey and a horse and between horse breeds. All ejaculates evaluated showed excellent semen motility characteristics, with significantly higher values for all kinematic parameters for donkeys compared with horses except for beat-cross frequency. Donkey sperm was faster and linear than the horse. Regarding horse breeds differences, the Spanish horse had higher average path velocity, curvilinear velocity, and beat-cross frequency compared with the Arabian horse. Spanish horse sperm was rapid, but Arab horse was more linear. The principal component analysis showed three sperm subpopulations in the ejaculate of donkeys and horses with a significantly different motility characteristic between them. The dominant subpopulation for both donkey and horse was for rapid, straight, and linear with a high beat sperm (38.2 and 41.7%, respectively), whereas the lowest subpopulation was for the slowest and non-linear sperms. This, plus slight differences in the distribution of these subpopulations between Arabian and Spanish horses, were found. In conclusion, higher frames permitted to have a new interpretation of motile subpopulations with species and breed differences. More so, future works on donkey and horse breed spermatozoa should take into account differences between breeds that may interfere and alter the real analysis performed.

Introducción. La movilidad es el indicador de calidad seminal más utilizado, por tanto, es necesario determinar una adecuada frecuencia de fotogramas (frame rate, FR) para estandarizar los análisis cinéticos con los equipos de análisis computadorizado de semen (CASA-Mot) y disminuir la variabilidad entre laboratorios, lo que optimizará el número de dosis seminales y su calidad para la industria porcina basada en inseminación artificial (IA). Objetivo. Determinar la frecuencia óptima de fotogramas sobre la cinética espermática de verraco mediante un sistema comercial CASA-Mot y analizar la distribución de subpoblaciones espermáticas con el FR óptimo. Materiales y métodos. Se utilizaron veinte dosis seminales de diez verracos Pietrain (Sus scrofa domestica). El periodo experimental fue entre febrero y julio del año 2017. Se utilizó el sistema CASA-Mot (ISAS®v1) con cámaras de recuento ISAS®D4C20 precalentadas a 37 °C. El tiempo de captura fue de dos segundos. Resultados. La frecuencia de fotogramas afectó los parámetros de cinética espermática, exceptuando la velocidad rectilínea (VSL), debido a que no representó un comportamiento dependiente del FR. Las demás variables presentaron alteraciones significativas al cambiar su FR, y la velocidad curvilínea (VCL) y velocidad promedio (VAP) fueron las variables más sensibles al cambio. Se obtuvieron tres componentes principales (PC) definidos por patrones de movilidad, progresividad y oscilación, en donde el PC1 (velocidad-ondulación) fue el más influyente, debido a que representó el 51,34 % y 55,08 % de la varianza total explicada para 25 y 200 fps (fotogramas por segundo), respectivamente. Conclusión. La frecuencia óptima de fotogramas para analizar variables de cinética espermática en verraco con un sistema CASA-Mot fue de 200 fps. Una tasa de fotogramas inferior provoca errores en la estimación de la trayectoria de los espermatozoides. La distribución de subpoblaciones espermáticas varía según la tasa de fotogramas utilizada. Es necesario establecer un protocolo de análisis para homogenizar los resultados.

There has been very limited use of computer assisted semen analysis (CASA) to evaluate reptile sperm. The aim of this study was to examine sperm kinematic variables in American crocodile (Crocodylus acutus) semen samples and to assess whether sperm subpopulations could be characterized. Eight ejaculates (two ejaculates/male) from four sexually mature captive crocodiles were obtained. An ISAS®v1 CASA-Mot system, with an image acquisition rate of 50 Hz, and ISAS®D4C20 counting chambers were used for sperm analyses. The percentages of motile and progressively motile spermatozoa did not differ among animals (P > 0.05) but there was a significant animal effect with regards to kinematic variables (P < 0.05). Principal component (PC) analysis revealed that kinematic variables grouped into three components: PC1, related to velocity; PC2 to progressiveness and PC3 to oscillation. Subpopulation structure analysis identified four groups (P < 0.05), which represented, on average, 9.8%, 32.1%, 26.8%, and 31.3% of the total sperm population. Males differed in the proportion of sperm in each of the kinematic subpopulations. This new approach for the analysis of reptile sperm kinematic subpopulations, reflecting quantifiable parameters generated by CASA system technology, opens up possibilities for future assessments of crocodile sperm and will be useful in the future development of assisted reproduction for these species.

El objetivo de este estudio fue evaluar la presencia de polimorfismos de nucleótido simple (SNP´s) en genes de µ-calpaína (CAPN-1) y calpastatina (CAST) asociados a atributos de terneza en carne bovina y relacionar la presencia de estos genes con las diferencias esperadas de progenie (DEP´s) para características de crecimiento. Se recolectaron muestras de sangre (n = 82) de machos reproductores Brahman de registro, pertenecientes a 7 haciendas, ubicadas en 4 regiones geográficas de Costa Rica. Todos los individuos se encontraban incluidos en el programa de mejoramiento genético de bovinos de carne, dirigido por la Corporación Ganadera (CORFOGA). Las muestras de sangre fueron analizadas en el laboratorio de biotecnología molecular del Instituto Tecnológico de Costa Rica, Sede San Carlos. Se analizó la presencia de los SNP´s CAPN316, CAPN4751 (gen CAPN-1) y CAST2959 (gen CAST). Se estimaron las frecuencias alélicas en CAPN316 para C (0,006), G (0,994) y genotipos CC (0,000), CG (0,013) y GG (0,987). El marcador CAPN4751 presentó frecuencias alélicas para C (0,165), T (0,835), así como frecuencias genotípicas de CC (0,025), CT (0,279) y TT (0,696). El polimorfismo de nucleótido simple CAST2959 presentó frecuencias alélicas para A (0,667), G (0,333), y genotípicas para AA (0,432), AG (0,469) y GG (0,099). Se observó una baja frecuencia del alelo favorable C del gen CAPN316, sin embargo, se observó una distribución de frecuencias alélicas más equilibrada en CAPN4751 y CAST2959. En el análisis de asociación entre SNP´s favorables y DEP´s, se encontraron asociaciones muy débiles o incluso ausencia de correlación.

The aim was to determine the relationship between kinematic parameters of boar spermatozoa and fertility rates of sow, as well as to assess the effect of sperm clusters on the fertility capacity of the ejaculate. Semen samples were collected from 11 sexually mature boars. Samples were analyzed by an ISAS®v1 CASA-Mot system for eight kinematic parameters. Ejaculate clusters were characterized using multivariate procedures, such as principal factors (PFs) analysis and clustering methods (the k-means model). Four different ejaculate clusters were identified from two kinematic PFs which involved linear trajectory and velocity. There were differences (p < 0.05) between the sperm kinematic variables by sire line. There was no statistical difference (p > 0.05) between dam lines and ejaculate clusters in fertility variables. The discriminant ability of the different kinematics of sperm variables to predict litter size fertility was analyzed using receiver operating characteristics (ROC) curve analysis. Curvilinear velocity (VCL), average path velocity (VAP), amplitude of lateral head displacement (ALH), and beat-cross frequency (BCF) showed significant, albeit limited, predictive capacity for litter size fertility variables (range: 0.55–0.58 area under curve, AUC). The kinematic analysis of the ejaculates in clusters did not have a predictive capacity for litter size variables.

Dogs have been under strong artificial selection as a consequence of their relationship with man. Differences between breeds are evident that could be reflected in seminal characteristics. The present study was to evaluate differences in sperm head morphometry between seven well-defined breeds of dog: the British Bulldog, Chihuahua, German Shepherd, Labrador Retriever, Spanish Mastiff, Staffordshire Terrier, and Valencian Rat Hunting dog. Semen samples were obtained by masturbation and smears stained with Diff-Quik. Morphometric analysis (CASA-Morph) produced four size and four shape parameters. Length, Ellipticity, and Elongation showed higher differences between breeds. MANOVA revealed differences among all breeds. Considering the whole dataset, principal component analysis (PCA) showed that PC1 was related to head shape and PC2 to size. Procluster analysis showed the British Bulldog to be the most isolated breed, followed by the German Shepherd. The PCA breed by breed showed the Chihuahua, Labrador Retriever, Spanish Mastiff, and Staffordshire Terrier to have PC1 related to shape and PC2 to size, whereas the British Bulldog, Valencia Rat Hunting dog, and German Shepherd had PC1 related to size and PC2 to shape. The dendrogram for cluster groupings and the distance between them showed the British Bulldog to be separated from the rest of the breeds. Future work on dog semen must take into account the large differences in the breeds′ sperm characteristics. The results provide a base for future work on phylogenetic and evolutionary studies of dogs, based on their seminal characteristics.

This work provides information on the blue fox ejaculated sperm quality needed for seminal dose calculations. Twenty semen samples, obtained by masturbation, were analyzed for kinematic and morphometric parameters by using CASA-Mot and CASA-Morph system and principal component (PC) analysis. For motility, eight kinematic parameters were evaluated, which were reduced to PC1, related to linear variables, and PC2, related to oscillatory movement. The whole population was divided into three independent subpopulations: SP1, fast cells with linear movement; SP2, slow cells and nonoscillatory motility; and SP3, medium speed cells and oscillatory movement. In almost all cases, the subpopulation distribution by animal was significantly different. Head morphology analysis generated four size and four shape parameters, which were reduced to PC1, related to size, and PC2, related to shape of the cells. Three morphometric subpopulations existed: SP1: large oval cells; SP2: medium size elongated cells; and SP3: small and short cells. The subpopulation distribution differed between animals. Combining the kinematic and morphometric datasets produced PC1, related to morphometric parameters, and PC2, related to kinematics, which generated four sperm subpopulations - SP1: high oscillatory motility, large and short heads; SP2: medium velocity with small and short heads; SP3: slow motion small and elongated cells; and SP4: high linear speed and large elongated cells. Subpopulation distribution was different in all animals. The establishment of sperm subpopulations from kinematic, morphometric, and combined variables not only improves the well-defined fox semen characteristics and offers a good conceptual basis for fertility and sperm preservation techniques in this species, but also opens the door to use this approach in other species, included humans.

The aim of the study was to compare the morphometric features of sperm head size and shape from the Pietrain line and the Duroc × Pietrain boar crossbred terminal lines, and to evaluate their relationship with reproductive success after artificial insemination of sows produced from crossbreeding the York, Landrace and Pietrain breeds. Semen samples were collected from 11 sexually mature boars. Only ejaculates with greater than 70% motility rate and ®v1 computer-assisted sperm analysis system for eight morphometric parameters of head shape and size (CASA-Morph). Sub-populations of morphometric ejaculates were characterized using multivariate procedures, such as principal component (PC) analysis and clustering methods (k-means model). Four different ejaculate sub-populations were identified from two PCs that involved the head shape and size of the spermatozoa. The discriminant ability of the different morphometric sperm variables to predict sow litter size was analyzed using a receiver operating characteristics (ROC) curve analysis. Sperm head length, ellipticity, elongation, and regularity showed significant predictive capacity on litter size (0.59, 0.59, 0.60, and 0.56 area under curve (AUC), respectively). The morphometric sperm sub-populations were not related to sow litter size.

This work evaluates sperm head morphometric characteristics in adolescents from 12 to 18 years of age, and the effect of varicocele. Volunteers between 150 and 224 months of age (mean 191, n = 87), who had reached oigarche by 12 years old, were recruited in the area of Barranquilla, Colombia. Morphometric analysis of sperm heads was performed with principal component (PC) and discriminant analysis. Combining seminal fluid and sperm parameters provided five PCs: two related to sperm morphometry, one to sperm motility, and two to seminal fluid components. Discriminant analysis on the morphometric results of varicocele and nonvaricocele groups did not provide a useful classification matrix. Of the semen-related PCs, the most explanatory (40%) was related to sperm motility. Two PCs, including sperm head elongation and size, were sufficient to evaluate sperm morphometric characteristics. Most of the morphometric variables were correlated with age, with an increase in size and decrease in the elongation of the sperm head. For head size, the entire sperm population could be divided into two morphometric subpopulations, SP1 and SP2, which did not change during adolescence. In general, for varicocele individuals, SP1 had larger and more elongated sperm heads than SP2, which had smaller and more elongated heads than in nonvaricocele men. In summary, sperm head morphometry assessed by CASA-Morph and multivariate cluster analysis provides a better comprehension of the ejaculate structure and possibly sperm function. Morphometric analysis provides much more information than data obtained from conventional semen analysis.

DNA fragmentation has been shown to be one of the causes of male infertility, particularly related to repeated abortions, and different methods have been developed to analyze it. In the present study, two commercial kits based on the SCD technique (Halosperm ® and SDFA) were evaluated by the use of the DNA fragmentation module of the ISAS ® v1 CASA system. Seven semen samples from volunteers were analyzed. To compare the results between techniques, the Kruskal-Wallis test was used. Data were used for calculation of Principal Components (two PCs were obtained), and subsequent subpopulations were identified using the Halo, Halo/Core Ratio, and PC data. Results from both kits were significantly different (P < 0.001). In each case, four subpopulations were obtained, independently of the classification method used. The distribution of subpopulations differed depending on the kit used. From the PC data, a discriminant analysis matrix was obtained and a good a posteriori classification was obtained (97.1% for Halosperm and 96.6% for SDFA). The present results are the first approach on morphometric evaluation of DNA fragmentation from the SCD technique. This approach could be used for the future definition of a classification matrix surpassing the current subjective evaluation of this important sperm factor.

Sperm quality is evaluated for the calculation of sperm dosage in artificial reproductive programs. The most common parameter used is motility, but morphology has a higher potential as a predictor of genetic quality. Morphometry calculations from CASA-Morph technology improve morphological evaluation and allow mathematical approaches to the problem. Semen from 28 Holstein bulls was collected by artificial vagina, and several ejaculates were studied. After general evaluation, samples were diluted, packaged in 0.25 ml straws, and stored in liquid nitrogen. Two straws per sample were thawed, and slides were processed and stained with Diff-Quik. Samples were analyzed by a CASA-Morph system for eight morphometric parameters. In addition to the "classical" statistical approach, based on variance analysis (revealing differences between animals, ejaculates, and straws), principal component (PC) analysis showed that the variables were grouped into PC1, related to size, and PC2 to shape. Subpopulation structure analysis showed four groups, namely, big, small, short, and narrow from their dominant characteristics, representing 31.0%, 27.3%, 24.1%, and 17.7% of the total population, respectively. The distributions varied between animals and ejaculates, but between straws, there were no differences in only four animals. This modern approach of considering an ejaculate sperm population as divided into subpopulations reflecting quantifiable parameters generated by CASA-Morph systems technology opens a new view on sperm function. This is the first study applying this approach to evaluate different ejaculates and straws from the same individual. More work must be done to improve seminal dose calculations in assisted reproductive programs.

In order to optimize the donkey sperm motility analysis by the CASA (Computer Assisted Sperm Analysis)-Mot system, twelve ejaculates were collected from six jackasses. Capillary loaded chamber (CLC), ISAS®D4C depths 10 and 20 µm, ISAS®D4C Leja 20 and drop displacement chamber (DDC), Spermtrack® (Spk) depths 10 and 20 µm were used. Sperm kinematic variables were evaluated using each chamber and a high-resolution camera capable of capturing a maximum of 500 frames/second (fps). The optimum frame rate (OFR) (defined according to curvilinear velocity—VCL) was dependent on chamber type. The highest OFR obtained was 278.46 fps by Spk20. Values for VCL, straight-line velocity (VSL), straightness (STR), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) were high in DDC and 10 µm depth. In both DDC 10 and 20 µm, the sperm velocities (VCL, VSL, VAP) and ALH values decreased significantly from the centre to the edges, while Wobble and BCF increased. No defined behavior was observed along the CLC. However, all the kinematic variables had a higher value in a highly concentrated sample, in both chamber types. In conclusion, analyzing a minimum of nine fields at 250 fps from the centre to the edges in Spk10 chamber using a dilution of 30 × 106 sperm/mL offers the best choice for donkey computerised sperm motility analysis.

The ejaculate is heterogenous and sperm sub-populations with different kinematic patterns can be identified in various species. Nevertheless, although these sub-populations are statistically well defined, the statistical differences are not always relevant. The aim of the present study was to characterize kinematic sub-populations in sperm from two bovine species, and diluted with different commercial extenders, and to determine the statistical relevance of sub-populations through Bayesian analysis. Semen from 10 bulls was evaluated after thawing. An ISAS®v1 computer-assisted sperm analysis (CASA)-Mot system was employed with an image acquisition rate of 50 Hz and ISAS®D4C20 counting chambers. Sub-populations of motile spermatozoa were characterized using multivariate procedures such as principal components (PCs) analysis and clustering methods (k-means model). Four different sperm sub-populations were identified from three PCs that involved progressiveness, velocity, and cell undulatory movement. The proportions of the different sperm sub-populations varied with the extender used and in the two species. Despite a statistical difference (p < 0.05) between extenders, the Bayesian analysis confirmed that only one of them (Triladyl®) presented relevant differences in kinematic patterns when compared with Tris-EY and OptiXcell®. Extenders differed in the proportion of sperm cells in each of the kinematic sub-populations. Similar patterns were identified in Bos taurus and Bos indicus. Bayesian results indicate that sub-populations SP1, SP2, and SP3 were different for PC criteria and these differences were relevant. For velocity, linearity, and progressiveness, the SP4 did not show a relevant difference regarding the other sperm sub-populations. The classical approach of clustering or sperm subpopulation thus may not have a direct biological meaning. Therefore, the biological relevance of sperm sub-populations needs to be reevaluated.

La evaluación de la calidad seminal constituye una herramienta para mejorar los índices reproductivos en las granjas porcinas. La movilidad del espermatozoide es un parámetro que se utiliza en la evaluación de la calidad espermática, porque se relaciona con el nivel de energía del gameto masculino. El objetivo de este estudio fue determinar la influencia de la composición racial sobre variables reproductivas de calidad, cinética y movilidad del semen en verracos. Durante el año 2016, se recolectaron 240 eyaculados de 63 verracos con una edad media de 24,4±10,9 meses. Se identificaron seis grupos raciales: Duroc (D), Yorkshire (Y), Landrace (L), F1 Pietrain*Duroc (PD) y dos líneas genéticas (LA y LB). Los grupos raciales con mayor volumen de eyaculado presentaron un número total de espermatozoides más elevado (P

Se evaluó el efecto de dos tipos de sonda (IC, intracervical, e IU, intrauterina) sobre variables de producción y reproducción en cerdas de primer parto y multíparas. Hubo un efecto (P < 0,05) del tipo de sonda (IU o IC) sobre los lechones destetados (11,61 ± 0,29 y 10,85 ± 0,33, respectivamente) y el peso de la camada al destete (78,28 ± 2,47 y 68,80 ± 2,85, respectivamente). Las hembras inseminadas con la sonda intrauterina presentaron valores mayores que las inseminadas con la sonda intracervical. Además, se encontró un efecto (P < 0,05) del orden de parto (multípara o de primer parto) sobre los lechones nacidos vivos, los lechones nacidos muertos (2,26 ± 0.32 y 1,45 ± 0,52, respectivamente), los lechones nacidos en total (14,34 ± 0,43 y 12,54 ± 0,68, respectivamente), el peso de la camada al nacer (16,11 ± 0,39 y 13,95 ± 0,62, respectivamente) y al destete (76,40 ± 2,01 y 70,68 ± 3,20, respectivamente). Las hembras multíparas presentaron valores mayores con respecto a las de primer parto. No hubo efecto (P > 0,05) de interacción tipo catéter-orden de parto sobre las variables analizadas.

Los sistemas computadorizados de análisis seminal (CASA, en inglés), se utilizan en reproducción animal conel propósito de valorar de forma rápida y fiable la calidad espermática. Actualmente, en los centros de inseminación artificial se debe disponer de sistemas fiables y objetivos que garanticen una confiabilidad de datos reproductivos.El objetivo fue realizar una revisión acerca de los sistemas de análisis seminal asistidos por computadora, enespecies ganaderas. El uso de sistemas CASA sustituye los análisis rutinarios que introducen sesgos y que conllevaa obtener distintos grados de imprecisión. Además, si se consideran las limitaciones intrínsecas de microscopíao espermatozoides con diferentes patrones de movilidad, lo más probable es que se introduzca una variabilidad considerable que repercute en la objetividad de la estimación. En un principio, este tipo de tecnologías eran pocoaccesibles, pero actualmente se utilizan en la mayoría de laboratorios para la evaluación de muestras espermáticas en distintas especies de interés zootécnico. Las técnicas computadorizadas son capaces de emitir informes con un número elevado de características, minimizar el factor subjetivo de los análisis rutinarios de semen y garantizar una mejor correlación con la capacidad fecundante de los espermatozoides, por medio de las relaciones entre la motilidad espermática y su competencia funcional. El uso de sistemas CASA junto con el análisis estadístico multivariante, han demostrado la existencia de subpoblaciones tanto cinéticas como morfométricas en eyaculados de diferentesespecies, desterrando la idea de que los eyaculados son homogéneos, formados por una única población espermáticacon distribución normal.

Los sistemas actuales de producción de carne bovina deben ser competitivos y estar orientados hacia la calidad cárnica como concepto de comercialización. El objetivo de este trabajo fue evaluar el componente genético y la suplementación en la fase final de novillos en pastoreo, sobre el crecimiento, composición de la canal y rendimiento cárnico. El periodo de evaluación fue de 126 días durante el año 2013, en la finca “La Vega” ubicada en el Cantón de San Carlos, Alajuela, Costa Rica. Se evaluó: en ganancia diaria de peso, espesor de grasa dorsal, marmoleo, profundidad de músculo, peso en pie, merma de transporte de finca a planta, peso canal caliente, rendimiento pie a canal, conformación muscular, grasa perirenal, diámetro de la pierna, longitud de la canal, área del ojo del lomo, grosor de grasa dorsal, pH y temperatura de la canal (2 y 24 h), merma de peso de canal caliente a canal fría. El componente genotípico de los bovinos incidió sobre las características de canal, así como el tipo de suplementación ofrecida. Los animales cruzados Brahman*Charolais (F1), presentaron mejor desempeño que los Brahman y/o Charolais, en características como profundidad de músculo y rendimiento en cortes cárnicos como lomito, posta de cuarto y punta de solomo. La suplementación ofrecida y el componente genotípico no influyeron en el rendimiento de carne vendible.

El objetivo del presente trabajo fue evaluar el efecto del componente racial y la suplementación energética en novillos en etapa de nalización en pastoreo sobre la ganancia de peso, la calidad de la canal y la fuerza de corte (kg). El periodo de evaluación del estudio fue de 144 días, durante el año 2014. Este se realizó en la nca “La Vega” ubicada en el cantón de San Carlos, Alajuela, Costa Rica. Se utilizaron veintiséis animales distribuidos en dos grupos raciales y tres niveles de suplementación para un total de seis tratamientos experimentales: E-T0 (3,2 Mcal), C-T0 (3,2 Mcal), E-T1 (3,4 Mcal), C-T1 (3,4 Mcal), E-T2 (3,67 Mcal) y C-T2 (3,67 Mcal). Se evaluaron: ganancia diaria de peso (GDP) y peso vivo y de la canal, marmoleo, desarrollo muscular, grasa dorsal, rendimiento en canal, espesor de grasa dorsal (EGD), peso de grasa perirenal, área del ojo del lomo (AOL) y nalmente la fuerza de corte (FC) como parámetro de calidad de carne. No se encontraron diferencias debido al efecto de la ración (P>0,05) para ganancia diaria de peso, peso de la canal, marmoleo, desarrollo muscular y grasa dorsal. Se encontró un efecto del componente racial sobre la calidad de carne (P

In the present study, the impact of non-functional spermatozoa on the cryopreservation success of functional boar spermatozoa was evaluated. Fifteen sperm-rich ejaculate fractions collected from five fertile boars were frozen with different proportions of induced non-functional sperm (0--native semen sample-, 25, 50 and 75% non-functional spermatozoa). After thawing, the recovery of motile and viable spermatozoa was assessed, and the functional of the spermatozoa was evaluated from plasma membrane fluidity and intracellular reactive oxygen species (ROS) generation upon exposure to capacitation conditions. In addition, the lipid peroxidation of the plasma membrane was assessed by the indirect measurement of malondialdehyde (MDA) generation. The normalized (with respect to a native semen sample) sperm motility (assessed by CASA) and viability (cytometrically assessed after staining with Hoechst 33342, propidium iodide and fluorescein-conjugated peanut agglutinin) decreased (p