Cryopreservation efficiency of red-rumped agouti (Dasyprocta leporina) sperm obtained from different origins through epididymal retrograde flushing or electroejaculation.

This study investigated whether the origin of sperm (epididymal vs. ejaculate) affects the cryopreservation efficiency in agouti (Dasyprocta leporina). Five sexually mature agoutis underwent electroejaculation, resulting in obtaining four semen samples. After 15 days, the same animals were euthanized, and through retrograde flushing, sperm samples were obtained from the epididymis tails. In both collection methods, samples were evaluated for sperm parameters (sperm concentration, motility, vigor, membrane integrity, osmotic response, and morphology). Then, samples were diluted in ACP 109c, added with 20% egg yolk, and a final concentration of 6% glycerol. Finally, the samples were packaged in 0.25 mL straws and frozen in liquid nitrogen. After one week, samples were thawed and evaluated in the same way as fresh samples, with the addition of membrane integrity analysis using fluorescent probes (C-FDA/PI) and computerized analysis (CASA). Immediately after obtaining the sperm, samples obtained directly from the epididymis presented higher values (P ≤ 0.05) than those obtained by electroejaculation concerning the parameters of volume, sperm concentration, and total number of sperm (1,398.25 ± 206.0 x106 and 184.5 ± 78.0 x106 sperm). On the other hand, in the classical evaluation of the other sperm parameters and the computerized analysis (CASA) after thawing, such as total motility, no statistical differences were observed between sperm from both origins (ejaculate: 16.7 ± 8.2% and epididymal: 24.8 ± 12.0%, P > 0.05). This demonstrates the possibility of direct application of the cryopreservation protocol for agouti (D. leporina) sperm obtained via the epididymis or ejaculate.

Microbiological load and preantral follicle preservation using different systems for ovarian tissue vitrification in the red-rumped agouti.

We evaluated the effect of open and closed systems used for ovarian tissue vitrification on the microbiological load and preservation of preantral follicles (PAFs) in the red-rumped agoutis. The ovaries from eight females were recovered and fragmented, with four cortexes fragments immediately fixed and evaluated (fresh group). The other fragments were processed for the solid-surface vitrification method (SSV) or an ovarian tissue cryosystem (OTC) using fetal calf serum, ethylene glycol, and sucrose as cryoprotectants, stored for two weeks, and rewarmed. Subsequently, fragments were subjected to a 24-h in vitro culture and assessed for microbiological load, PAF morphology, and DNA integrity. There was no fungal contamination; however, the vitrified samples from two individuals showed bacterial contamination of 79 200 colony forming units per milliliter (CFU)/mL for SSV and 3120 CFU/mL for OTC. From those samples, a total of eight different types of bacterial colonies were isolated and identified as coagulase-negative Staphylococci and Gram-positive bacilli. Regarding PAF morphology, both systems provided adequate preservation, with values higher than 70% normal follicles observed before and after culture. The TUNEL assay revealed that both SSV (52.39%) and OTC (41.67%) could preserve DNA integrity after vitrification and after 24 h of culture. In summary, both open and closed systems were equally efficient in preserving agouti ovarian tissues, especially concerning the preantral follicle morphology and DNA integrity; however, the OTC seems to provide a less adequate environment for bacterial proliferation.

Cryopreservation and Culture of Testicular Tissues: An Essential Tool for Biodiversity Preservation.

Systematic cryo-banking of reproductive tissues could enhance reproductive management and ensure sustainability of rare mammalian genotypes. Testicular tissues contain a vast number of germ cells, including at early stages (spermatogonia and spermatocytes), that can potentially develop into viable spermatozoa after grafting or culture in vitro, and the resulting sperm cells then can be used for assisted reproductive techniques. The objective of this review was to describe current advances, limitations, and perspectives related to the use of testicular tissue preservation as a strategy for the conservation of male fertility. Testes can be obtained from mature or prepubertal individuals, immediately postmortem or by orchiectomy, but testicular biopsies could also be an alternative to collect samples from living individuals. Testicular fragments can be then cryopreserved by using slow or ultra-rapid freezing, or even vitrification methods. The composition of cryopreservation media can vary according to species-specific characteristics, especially regarding the cryoprotectant type and concentration. Finally, spermatozoa have been usually obtained after xenografting of testicular fragments into severely immunodeficient mice, while this method still has to be optimized after in vitro culture conditions.

Influence of different extenders on morphological and functional parameters of frozen-thawed spermatozoa of jaguar (Panthera onca).

Due to the global decrease in jaguar population, conservation strategies are essential and the development of effective semen cryopreservation protocols would contribute to the formation of germplasm banks. Therefore, the objectives were to (1) evaluate the use of TRIS and ACP-117c extenders for jaguar semen freezing, (2) describe the ultrastructural changes in sperm after cryopreservation, and (3) evaluate the binding capacity of the thawed sperm. Eight ejaculates from five mature individuals were collected by electroejaculation, extended in TRIS or a coconut based-extender (ACP-117c), and frozen in liquid nitrogen. Samples were evaluated for sperm motility, vigor, membrane functionality, mitochondrial activity, morphology (using light microscopy, scanning electron microscopy - SEM and transmission electron microscopy - TEM), sperm kinetic parameters (by computerized analysis - CASA), and sperm binding capability using an egg yolk perivitelline membrane assay. Samples preserved in TRIS presented better post-thaw motility (46.0 ±â€¯7.7%) and membrane functionality (60.5 ±â€¯4.2%) and higher mitochondrial activity (21.5 ±â€¯3.7%) than those preserved in ACP-117c (20.9 ±â€¯5.4% motile sperm; 47.1 ±â€¯2.5% functional membrane; 11.8 ±â€¯1.7% mitochondrial activity). Regarding ultrastructural evaluations, SEM showed that both extenders were able to preserve the superficial membrane of the sperm, but TEM revealed the occurrence of nuclear electron lucent points, especially in samples extended in ACP-117c. Additionally, TRIS also provided a higher number of sperm bound to the perivitelline membrane (29.5 ±â€¯3.3%) in comparison to samples diluted in ACP-117c (18.6 ±â€¯1.5%). Overall, we suggest the use of a TRIS-based extender for cryopreservation of jaguar semen.

Establishment of a protocol for the isolation of ovarian preantral follicles derived from collared peccaries (Pecari tajacu).

We compare the efficiency of mechanical or enzymatic methods, and their combination, for the isolation of ovarian preantral follicles (PFs) from collared peccaries. The ovaries from six females were subjected to the different methods investigated here. For the enzymatic method, ovary fragments were exposed to collagenase type IV in TCM-HEPES medium; the mechanical procedure was based on ovarian cortex dissociation by using a scalpel blade. The residual solution obtained after the mechanical isolation was subjected to the enzymatic procedure. The number of isolated PFs was quantified and classified as primordial, primary, or secondary; their viability was assessed using trypan blue dye assay. To confirm the results, PFs derived from the most efficient method were evaluated for integrity using scanning electron microscopy (SEM) and subjected to a 24 h in vitro culture for subsequent evaluation of viability by using fluorescent probes. A higher number of PFs (P < 0.05) was obtained from the enzymatic method (961.7 ± 132.9) in comparison with the mechanical method (434.3 ± 88.9), but no difference was observed between the two methods and their combination (743.2 ± 92.8). The trypan blue assay showed that the enzymatic method (98.7 ± 0.6%) provided the highest percentage of viable follicles (P < 0.05). Furthermore, SEM confirmed the ultrastructural integrity of the surface architecture of peccary PFs isolated by the enzymatic procedure; epifluorescence microscopy was used to confirm their viability (86.0%). In conclusion, we suggest that the enzymatic method investigated here is useful for the isolation of viable ovarian PFs from collared peccaries.

Estrous synchronization in captive collared peccaries (Pecari tajacu) using a prostaglandin F2α analog.

We verify the efficiency of a protocol for estrus synchronization in captive female collared peccaries (Pecaricari tajacu) using the prostaglandin analog D-cloprostenol. Five adult female collared peccaries received an intramuscular administration of 60 µg D-cloprostenol, which procedure was repeated after a 9-day interval. For 10 days after second the D-cloprostenol administration, females were monitored for changes in external genitalia, ovarian ultrasonography, vaginal cytology and reproductive hormonal dosage. As a result, four females synchronized their estrous at 9.5 ± 0.5 days after the second administration of the prostaglandin analog. Such females showed external signs of estrus, including vulvar opening, hyperemic vaginal mucosa, and vaginal mucus, concomitant with an increase in the proportion of superficial cells (52.2 ± 9.9%) verified through vaginal cytology. An estrogen peak of 22.7 ± 3.4 pg/ml was detected by hormonal dosage, and the presence of anechoic follicles measuring 0.29 ± 0.05 × 0.32 ± 0.07 mm were detected in the ovary by ultrasonography. Given these findings, we suggest that D-cloprostenol may be effective for use in estrus synchronization in collared peccaries.

Relações entre a câmara de Neubauer a espectrofotometria utilizadas para a determinação da concentração espermática de catetos (Pecari tajacu) / Relationship between Neubauer counting chamber and spectrophotometer used for the sperm concentration determination in collared peccaries (Pecari tajacu)

Em ejaculados provenientes de 28 catetos, verificou-se a existência de relações entre a concentração espermática determinada por meio da câmara de Neubauer e a tramitância observada por espectrofotometria, utilizando comprimentos de onda variando de 530 a 590nm. Os ejaculados apresentaram uma concentração média de 283,9±30,8x106 espermatozoides mL-1, com variação de 30 a 640x106 espermatozoides mL-1. Os valores para tramitância variaram entre 36,9 a 96,3, nos diferentes comprimentos de onda. Não foram detectadas relações significativas entre os dois métodos (P>0,05). Dessa forma, não se recomenda a espectrofotometria para os procedimentos de rotina na determinação da concentração espermática em catetos.

In ejaculates derived from 28 collared peccaries, we verified the existence of relationships between sperm concentration determined by the Neubauer counting chamber and the tramitance verified by spectrophotometer, under wavelengths varying from 530 to 590nm. Ejaculates presented a concentration of 283.9±30.8x106sperm m-1, varying from 30 to 640x106sperm mL-1. Values for tramitance varied from 36.9 to 96.3, under different wavelengths. No significant relationship was verified between two methods (P>0.05). Thus, the spectrophotometer is not recommended for routine procedures of sperm concentration measurement in collared peccaries.

Educador social: Imagem e relações com crianças em situação de acolhimento institucional / Social educator: Image and relationship with children in shelter institutions / Educador social: La imagen y relaciones con los niños en situaciones de refugio / Éducateur Social: Image et relations avec les enfants dans les situations de abri

Este estudo teve como objetivo identificar a imagem que educadores sociais têm da criança em situação de acolhimento institucional e relacioná-la com as ações propostas por estes em espaços de convivência institucional. A investigação ocorreu em uma instituição abrigo (Diadema / SP) que atende crianças vítimas de negligência e / ou maus tratos. Os instrumentos utilizados foram: Teste do Desenho Temático e Observação. A técnica foi aplicada individualmente nas seis educadoras que integravam a instituição. Já a observação foi realizada nos momentos nos quais havia uma única educadora acompanhando o grupo de crianças em atividades de rotina do abrigo. Os dados obtidos com cada uma das educadoras foram analisados separadamente e posteriormente integrados aos registros da Observação. Na continuidade, uma síntese conclusiva foi elaborada para cada uma das participantes do estudo. Os resultados denotam falta de energia vital e ausência de contato afetivo com as crianças. Observa-se a predominância de educadoras com uma concepção do abrigo como um lugar provisório e inapropriado para o desenvolvimento da criança e descomprometidas com a tarefa de educá-las. O ambiente institucional parece favorecer a emergência de sentimentos de insegurança e inadequação e a ausência de perspectivas positivas para o futuro das crianças abrigadas. Esses sentimentos geram ansiedade, da qual se defendem por meio do uso de racionalização e do excessivo controle sobre o comportamento das crianças, desse modo impossibilitando uma interação mais favorável. As educadoras, em sua relação com as crianças, se configuram em modelos de identificação que, se bem elaborados, apontariam para uma instituição mais criativa, educativa e saudável. (AU)

This study aimed to identify the image that social educators have about children in shelter institutions and relate it to their proposed actions for institutional coexistence spaces. The research took place at a shelter institution (Diadema / SP) that works with children victims of negligence and / or abuse. The techniques used were Thematic Drawing Test and Observation. The technique was applied individually in six educators who worked for the institution. The observation was conducted in moments when only one educator was accompanying the group of children in routine activities in the shelter. The data obtained with each educator were analyzed separately and subsequently incorporated in the observation registries. In continuity, a conclusive synthesis was elaborated for each one of the study participants. The results show a lack of vital energy and emotional contact with the children. There is a predominance of educators with an image of the shelter as a temporary place, inappropriate for a child's development, and not committed to the task of children's education. The institutional environment seems to favor the emergency of insecurity feelings, inadequacy and lack of positive prospects for the future of the sheltered children. These feelings create anxiety, from which they try to protect themselves using rationalization and excessive control over the children's behavior, thus preventing a more favorable interaction. The educators, in their relationship with the children, configure themselves in models of identification that, if well developed, could point to a more creative, educational and healthy institution. (AU)

Este estudio tuvo como objetivo identificar la imagen que los educadores sociales tienen del niño en un albergue y se refieren a las acciones propuestas por ellos en los espacios de vida institucional. La investigación se llevó a cabo en una institución refugio (Diadema / SP), que sirve a los niños víctimas de abandono y / o maltrato. Los instrumentos utilizados fueron: Test de Dibujo y Nota temática. La técnica se aplicó de forma individual en seis educadores que formaban parte de la institución. Puesto que la observación se realizó en momentos en que sólo había un profesor que acompaña al grupo de los niños en las actividades rutinarias de la vivienda. Los datos obtenidos por cada uno de los maestros se analizaron por separado, seguido de un análisis de la integración de la producción gráfica de la Nota de los registros. Más tarde, una conclusión como resumen fue preparado para cada uno de los participantes en el estudio. Los resultados muestran una falta de energía vital y la falta de contacto emocional con los niños. Tenga en cuenta el predominio de los maestros con un dibujo de la vivienda como un hogar temporal e inadecuado para el desarrollo del niño, aparecen comprometidos en la tarea de educarlos. El ambiente institucional parecía fomentar sentimientos de inseguridad, la insuficiencia y falta de perspectivas positivas para el futuro de los niños protegidos. Estos sentimientos generan ansiedad para los que se defienden mediante el uso de la racionalización y del control excesivo sobre el comportamiento de los niños, imposibilitando una interacción más favorable. Los educadores en su relación con los niños, se han configurado para identificar los modelos que, si están bien elaboraos, apuntan a una institución más creativa, educativa y saludable. (AU)

Cette étude visait à identifier l'image que les éducateurs sociaux que l'enfant dans un abri et de le relier aux actions proposées par ceux qui dans les espaces de vie institutionnels. La recherche a eu lieu dans un établissement sous (Diadema / SP), qui sert aux enfants victimes de négligence et / ou de mauvais traitements. Les instruments utilisés étaient: test du dessin et de la note thématique. La technique a été appliquée individuellement dans six éducateurs qui faisaient partie de l'institution. Depuis l'observation a été effectuée à des moments où il n'y avait qu'un seul enseignant accompagnant le groupe d'enfants dans les activités de routine de l'abri. Les données obtenues par chacun des enseignants ont été analysés séparément, suivie d'une analyse intégrant la production graphique de la note dossiers. Plus tard, un résumé a été préparé pour la conclusion de chacun des participants à l'étude. Les résultats montrent un manque d'énergie vitale et le manque de contact émotionnel avec les enfants. Notez la prédominance des enseignants avec une conception de l'abri comme un lieu temporaire et inappropriée pour le développement de l'enfant, semblent ne pas s'engager dans la tâche de les éduquer. L'environnement institutionnel semblait susciter des sentiments d'insécurité, l'insuffisance et le manque de perspectives positives pour l'avenir des enfants protégés. Ces sentiments d'anxiété pour lequel ils se défendre grâce à l'utilisation de la rationalisation et un contrôle excessif sur le comportement des enfants, ce qui empêche une interaction plus favorable. Les éducateurs dans leur relation avec les enfants, sont configurés pour identifier les modèles qui, s'ils sont bien conçus, rappelle dans un établissement de plus de créativité, de l'éducation et la santé. (AU)

Educador social: imagem e relações com crianças em situação de acolhimento institucional / Social educator: image and relationship with children in shelter institutions / Educador social: la imagen y relaciones con los niños en situaciones de refugio / Éducateur social: image et relations avec les enfants dans les situations de abri

Este estudo teve como objetivo identificar a imagem que educadores sociais têm da criança em situação de acolhimento institucional e relacioná-la com as ações propostas por estes em espaços de convivência institucional. A investigação ocorreu em uma instituição abrigo (Diadema / SP) que atende crianças vítimas de negligência e / ou maus tratos. Os instrumentos utilizados foram: Teste do Desenho Temático e Observação. A técnica foi aplicada individualmente nas seis educadoras que integravam a instituição. Já a observação foi realizada nos momentos nos quais havia uma única educadora acompanhando o grupo de crianças em atividades de rotina do abrigo. Os dados obtidos com cada uma das educadoras foram analisados separadamente e posteriormente integrados aos registros da Observação. Na continuidade, uma síntese conclusiva foi elaborada para cada uma das participantes do estudo. Os resultados denotam falta de energia vital e ausência de contato afetivo com as crianças. Observa-se a predominância de educadoras com uma concepção do abrigo como um lugar provisório e inapropriado para o desenvolvimento da criança e descomprometidas com a tarefa de educá-las...

This study aimed to identify the image that social educators have about children in shelter institutions and relate it to their proposed actions for institutional coexistence spaces. The research took place at a shelter institution (Diadema / SP) that works with children victims of negligence and / or abuse. The techniques used were Thematic Drawing Test and Observation. The technique was applied individually in six educators who worked for the institution. The observation was conducted in moments when only one educator was accompanying the group of children in routine activities in the shelter. The data obtained with each educator were analyzed separately and subsequently incorporated in the observation registries. In continuity, a conclusive synthesis was elaborated for each one of the study participants. The results show a lack of vital energy and emotional contact with the children. There is a predominance of educators with an image of the shelter as a temporary place, inappropriate for a child's development, and not committed to the task of children's education...

Este estudio tuvo como objetivo identificar la imagen que los educadores sociales tienen del niño en un albergue y se refieren a las acciones propuestas por ellos en los espacios de vida institucional. La investigación se llevó a cabo en una institución refugio (Diadema / SP), que sirve a los niños víctimas de abandono y / o maltrato. Los instrumentos utilizados fueron: Test de Dibujo y Nota temática. La técnica se aplicó de forma individual en seis educadores que formaban parte de la institución. Puesto que la observación se realizó en momentos en que sólo había un profesor que acompaña al grupo de los niños en las actividades rutinarias de la vivienda. Los datos obtenidos por cada uno de los maestros se analizaron por separado, seguido de un análisis de la integración de la producción gráfica de la Nota de los registros. Más tarde, una conclusión como resumen fue preparado para cada uno de los participantes en el estudio. Los resultados muestran una falta de energía vital y la falta de contacto emocional con los niños. Tenga en cuenta el predominio de los maestros con un dibujo de la vivienda como un hogar temporal e inadecuado para el desarrollo del niño, aparecen comprometidos en la tarea de educarlos...

Cette étude visait à identifier l'image que les éducateurs sociaux que l'enfant dans un abri et de le relier aux actions proposées par ceux qui dans les espaces de vie institutionnels. La recherche a eu lieu dans un établissement sous (Diadema / SP), qui sert aux enfants victimes de négligence et / ou de mauvais traitements. Les instruments utilisés étaient: test du dessin et de la note thématique. La technique a été appliquée individuellement dans six éducateurs qui faisaient partie de l'institution. Depuis l'observation a été effectuée à des moments où il n'y avait qu'un seul enseignant accompagnant le groupe d'enfants dans les activités de routine de l'abri. Les données obtenues par chacun des enseignants ont été analysés séparément, suivie d'une analyse intégrant la production graphique de la note dossiers. Plus tard, un résumé a été préparé pour la conclusion de chacun des participants à l'étude. Les résultats montrent un manque d'énergie vitale et le manque de contact émotionnel avec les enfants. Notez la prédominance des enseignants avec une conception de l'abri comme un lieu temporaire et inappropriée pour le développement de l'enfant, semblent ne pas s'engager dans la tâche de les éduquer...